1: Pol J Vet Sci. 2008;11(4):411-4. Current issues connected with usage of genetically modified crops in production of feed and livestock feeding. Kwiatek K, Mazur M, Sieradzki Z. Department of Hygiene of Animal Feedingstuffs, National Veterinary Research Institute in Pulawy, Al. Partyzantów 57, 24-100 Puławy, Poland. kwiatekk@piwet.pulawy.pl Progress, which is brought by new advances in modern molecular biology, allowed interference in the genome of live organisms and gene manipulation. Introducing new genes to the recipient organism enables to give them new features, absent before. Continuous increase in the area of the biotech crops triggers continuous discussion about safety of genetically modified (GM) crops, including food and feed derived from them. Important issue connected with cultivation of genetically modified crops is a horizontal gene transfer and a bacterial antibiotic resistance. Discussion about safety of GM crops concerns also food allergies caused by eating genetically modified food. The problem of genetic modifications of GM crops used for livestock feeding is widely discussed, taking into account Polish feed law. PMID: 19227143 [PubMed - in process] 2: Anal Bioanal Chem. 2009 Feb 19. [Epub ahead of print] Sensitive and highly specific quantitative real-time PCR and ELISA for recording a potential transfer of novel DNA and Cry1Ab protein from feed into bovine milk. Guertler P, Paul V, Albrecht C, Meyer HH. Physiology Weihenstephan, Technische Universität München, Weihenstephaner Berg 3, 85350, Freising, Germany, patrick.guertler@wzw.tum.de. To address food safety concerns of the public regarding the potential transfer of recombinant DNA (cry1Ab) and protein (Cry1Ab) into the milk of cows fed genetically modified maize (MON810), a highly specific and sensitive quantitative real-time PCR (qPCR) and an ELISA were developed for monitoring suspicious presence of novel DNA and Cry1Ab protein in bovine milk. The developed assays were validated according to the assay validation criteria specified in the European Commission Decision 2002/657/EC. The detection limit and detection capability of the qPCR and ELISA were 100 copies of cry1Ab muL(-1) milk and 0.4 ng mL(-1) Cry1Ab, respectively. Recovery rates of 84.9% (DNA) and 97% (protein) and low (<15%) imprecision revealed the reliable and accurate estimations. A specific qPCR amplification and use of a specific antibody in ELISA ascertained the high specificity of the assays. Using these assays for 90 milk samples collected from cows fed either transgenic (n = 8) or non-transgenic (n = 7) rations for 6 months, neither cry1Ab nor Cry1Ab protein were detected in any analyzed sample at the assay detection limits. PMID: 19225766 [PubMed - as supplied by publisher] 3: J Agric Food Chem. 2009 Feb 13. [Epub ahead of print] Safety Assessment and Detection Method of Genetically Modified Chinese Kale ( Brassica oleracea cv. alboglabra ). Lin CH, Lu CT, Lin HT, Pan TM. Institute of Microbiology and Biochemistry, College of Life Science, National Taiwan University, Number 1, Section 4, Roosevelt Road, Taipei 10617, Taiwan, and Department of Food Science, Nutrition, and Nutraceutical Biotechnology, Shih Chien University, Number 70, Ta-Chih Street, Taipei 10462, Taiwan. Sporamins are tuberous storage proteins and account for 80% of soluble protein in sweet potato tubers with trypsin-inhibitory activity. The expression of sporamin protein in transgenic Chinese kale (line BoA 3-1) conferred insecticidal activity toward corn earworm [ Helicoverpa armigera (Hubner)] in a previous report. In this study, we present a preliminary safety assessment of transgenic Chinese kale BoA 3-1. Bioinformatic and simulated gastric fluid (SGF) analyses were performed to evaluate the allergenicity of sporamin protein. The substantial equivalence between transgenic Chinese kale and its wild-type host has been demonstrated by the comparison of important constituents. A reliable real-time polymerase chain reaction (PCR) detection method was also developed to control sample quality. Despite the results of most evaluations in this study being negative, the safety of sporamin in transgenic Chinese kale BoA 3-1 was uncluded because of the allergenic risk revealed by bioinformatic analysis. PMID: 19216530 [PubMed - as supplied by publisher] 4: Methods Mol Biol. 2009;483:69-87. Production and localization of recombinant pharmaceuticals in transgenic seeds. Rademacher T, Arcalis E, Stoger E. Institute for Molecular Biology, RWTH Aachen, Worringerweg, Aachen, Germany. Among the many plant-based production systems that have been developed for pharmaceutical proteins, seeds have the useful advantage of accumulating proteins in a relatively small volume, and recombinant proteins are very stable in dry seeds allowing long-term storage and facilitating distribution before processing.To take full advantage of the natural ability of endosperm cells to store large amounts of protein in a protected subcellular environment, it is useful to target recombinant proteins to appropriate storage organelles. In this chapter, we describe the distinct types of protein storage organelles in the cereal endosperm and a protocol for the detection of recombinant proteins in these organelles by immunofluorescence and immunogold labelling.The use of food and feed crops for the production of pharmaceutical proteins such as edible vaccines implies the need for strict separation of the transgenic seeds from the food and feed chain. For improved traceability visual markers may be co-expressed with the gene of interest in engineered seeds. DsRed is one example for a fluorescent protein that can be detected with high sensitivity using low tech equipment. We therefore describe the generation of transgenic maize plants expressing DsRed in a constitutive manner, and we point out the advantages of using this marker during the process of transformation and selection of plant tissue and later during breeding of transgenic lines into elite germplasm. PMID: 19183894 [PubMed - indexed for MEDLINE] 5: Mod Healthc. 2009 Jan 12;39(2):17. Battling the clones. CHW wants to avoid genetically altered foods. Rhea S. Publication Types: News PMID: 19172933 [PubMed - indexed for MEDLINE] 6: J Dairy Sci. 2009 Feb;92(2):444-57. Fate of lysostaphin in milk from individual cows through pasteurization and cheesemaking. Van Hekken DL, Wall RJ, Somkuti GA, Powell MA, Tunick MH, Tomasula PM. Dairy Processing and Products Research Unit, USDA, Agricultural Research Service, Eastern Regional Research Center, Wyndmoor, PA 19038, USA. diane.vanhekken@ars.usda.gov Transgenic cows secreting over 3 microg of lysostaphin/ mL of milk are protected against mastitis caused by Staphylococcus aureus, but it is unknown if active lysostaphin persists through dairy processing procedures or affects the production of fermented dairy foods. The objective of this study was to determine the fate of lysostaphin as milk was pasteurized and then processed into cheese. Raw milk from transgenic cows was heat treated at 63 degrees C for 30 min, 72 degrees C for 15 s (high temperature, short time), or 140 degrees C for 2 s (UHT). Portions of the high temperature, short-time milk were manufactured into semi-hard cheeses. Aliquots taken at each processing step were assayed to determine the quantity (ELISA) and activity (ability to inhibit S. aureus growth) of lysostaphin. Results indicated that most of the lysostaphin was present in the aqueous portion of the milk and was not affected by pasteurization, although UHT treatment reduced enzyme concentration by 60%. The quantity and activity of the lysostaphin decreased during cheesemaking. Based on the amount of lysostaphin present in the starting cheesemilk, 10 to 15% of the lysostaphin was recovered in the whey, 21 to 55% in the cheese curd at d 1, and 21 to 36% in cheese stored at 4 degrees C for 90 d. Enough of the lysostaphin secreted into milk by transgenic cows survived typical dairy processing conditions to impart potential value as a bioprotective agent against staphylococci in dairy foods. PMID: 19164654 [PubMed - indexed for MEDLINE] 7: J Genet Genomics. 2009 Jan;36(1):41-9. Index selection on seed traits under direct, cytoplasmic and maternal effects in multiple environments. Zhang W, Xu H, Zhu J. Institute of Bioinformatics, College of Agriculture and Biotechnology, Zhejiang University, Hangzhou 310029, China. Crop seeds are important sources of protein, oil, and carbohydrates for food, animal feeds, and industrial products. Recently, much attention has been paid to quality and functional properties of crop seeds. However, seed traits possess some distinct genetic characteristics in comparison with plant traits, which increase the difficulty of genetically improving these traits. In this study, diallel analysis for seed models with genotype by environment interaction (GE) effect was applied to estimate the variance-covariance components of seed traits. Mixed linear model approaches were used to estimate the genetic covariances between pair-wise seed and plant traits. The breeding values (BV) were divided into two categories for the seed models. The first category of BV was defined as the combination of direct additive, cytoplasmic, and maternal additive effects, which should be utilized for selecting stable cultivars over multi-environments. The three genetic effects, together with their GE interaction, were included in the second category of BV for selecting special lines to be grown in specific ecosystems. Accordingly, two types of selection indices for seed traits, i.e., general selection index and interaction selection index, were developed and constructed on the first and the second category BV, respectively. These proposed selection indices can be applied to solve the difficult task of simultaneously improving multiple seed traits in various environments. Data of crop seeds with regard to four seed traits and four yield traits based on the modified diallel crosses in Upland cotton (Gossypium hirsutum L.) were used as an example for demonstrating the proposed methodology. Publication Types: Research Support, Non-U.S. Gov't PMID: 19161944 [PubMed - in process] 8: Rev Med Suisse. 2008 Dec 10;4(183):2709. [Europe does not want to eat cloned food] [Article in French] Nau JY. jynau@orange.fr PMID: 19157291 [PubMed - indexed for MEDLINE] 9: Anal Chim Acta. 2009 Feb 16;634(1):75-82. Epub 2008 Dec 6. Evaluation of stable isotope labelling strategies for the quantitation of CP4 EPSPS in genetically modified soya. Ocaña MF, Fraser PD, Patel RK, Halket JM, Bramley PM. Centre for Chemical and Bioanalytical Sciences, Royal Holloway, University of London, Egham TW20 0EX, UK. The introduction of genetically modified (GM) crops into the market has raised a general alertness relating to the control and safety of foods. The applicability of protein separation hyphenated to mass spectrometry to identify the bacterial enolpyruvylshikimate-3-phosphate synthase (CP4 EPSPS) protein expressed in GM crops has been previously reported [M.F. Ocana, P.D. Fraser, R.K.P. Patel, J.M. Halket, P.M. Bramley, Rapid Commun. Mass Spectrom. 21 (2007) 319.]. Herein, we investigate the suitability of two strategies that employ heavy stable isotopes, i.e. AQUA and iTRAQ, to quantify different levels of CP4 EPSPS in up to four GM preparations. Both quantification strategies showed potential to determine whether the presence of GM material is above the limits established by the European Union. The AQUA quantification procedure involved protein solubilisation/fractionation and subsequent separation using SDS-PAGE. A segment of the gel in which the protein of interest was located was excised, the stable isotope labeled peptide added at a known concentration and proteolytic digestion initiated. Following recovery of the peptides, on-line separation and detection using LC-MS was carried out. A similar approach was used for the iTRAQ workflow with the exception that proteins were digested in solution and generated tryptic peptides were chemically tagged. Both procedures demonstrated the potential for quantitative detection at 0.5% (w/w) GM soya which is a level below the current European Union's threshold for food-labelling. In this context, a comparison between the two procedures is provided within the present study. Publication Types: Evaluation Studies Research Support, Non-U.S. Gov't PMID: 19154813 [PubMed - indexed for MEDLINE] 10: Recent Pat DNA Gene Seq. 2009;3(1):53-62. Current patents and future development underlying marker-assisted breeding in major grain crops. Utomo HS, Linscombe SD. Rice Research Station, Louisiana State University Agricultural Center, 1373 Caffey Rd., Rayne, Louisiana 70578, USA. hutomo@agcenter.lsu.edu Genomics and molecular markers provide new tools to assemble and mobilize important traits from different genetic backgrounds, including breeding lines and cultivars from different parts of the world and their related wild ancestors, to improve the quality and yield of the existing commercial cultivars to meet the increasing challenges of global food demand. The basic techniques of marker-assisted breeding, such as isolating DNA, amplifying DNA of interest using publicly available primers, and visualizing DNA fragments using standard polyacrylamid gel, have been described in the literature and, therefore, are available to scientists and breeders without any restrictions. A more sophisticated high-throughput system that includes proprietary chemicals and reagents, parts and equipments, software, and methods or processes, has been a subject of intensive patents and trade secrets. The high-throughput systems offer a more efficient way to discover associated QTLs for traits of economic importance. Therefore, an increasing number of patents of highly valued genes and QTLs is expected. This paper will discuss and review current patents associated with genes and QTLs utilized in marker-assisted breeding in major grain crops. The availability of molecular markers for important agronomic traits combined with more efficient marker detection systems will help reach the full benefit of MAS in the breeding effort to reassemble potential genes and recapture critical genes among the breeding lines that were lost during domestication to help boost crop production worldwide. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 19149739 [PubMed - indexed for MEDLINE] 11: Kennedy Inst Ethics J. 2008 Dec;18(4):393-402. FDA releases draft guidance on regulation of genetically engineered animals. Gluck JP, Holdsworth MT. Department of Psychology, University of New Mexico, Albuquerque, USA. PMID: 19143411 [PubMed - indexed for MEDLINE] 12: Eur J Histochem. 2008 Oct-Dec;52(4):263-7. Can a genetically-modified organism-containing diet influence embryo development? A preliminary study on pre-implantation mouse embryos. Cisterna B, Flach F, Vecchio L, Barabino SM, Battistelli S, Martin TE, Malatesta M, Biggiogera M. Dipartimento di Biologia Animale, Laboratorio di Biologia Cellulare e Neurobiologia, ed Instituto di Genetica Molecolare del CNR, University of Pavia, Italy. In eukaryotic cells, pre-mRNAs undergo several transformation steps to generate mature mRNAs. Recent studies have demonstrated that a diet containing a genetically modified (GM) soybean can induce modifications of nuclear constituents involved in RNA processing in some tissues of young, adult and old mice. On this basis, we have investigated the ultrastructural and immunocytochemical features of pre-implantation embryos from mice fed either GM or non- GM soybean in order to verify whether the parental diet can affect the morpho-functional development of the embryonic ribonucleoprotein structural constituents involved in pre-mRNA pathways. Morphological observations revealed that the general aspect of embryo nuclear components is similar in the two experimental groups. However, immunocytochemical and in situ hybridization results suggest a temporary decrease of pre-mRNA transcription and splicing in 2-cell embryos and a resumption in 4-8-cell embryos from mice fed GM soybean; moreover, pre-mRNA maturation seems to be less efficient in both 2-cell and 4-8-cell embryos from GM-fed mice than in controls. Although our results are still preliminary and limited to the pre-implantation phases, the results of this study encourage deepening on the effects of food components and/or contaminants on embryo development. Publication Types: Research Support, Non-U.S. Gov't PMID: 19109102 [PubMed - indexed for MEDLINE] 13: J Agric Food Chem. 2009 Jan 28;57(2):395-402. Event-specific detection of stacked genetically modified maize Bt11 x GA21 by UP-M-PCR and real-time PCR. Xu W, Yuan Y, Luo Y, Bai W, Zhang C, Huang K. Laboratory of Food Safety and Molecular Biology, College of Food Science and Nutritional Engineering, China Agricultural University, Beijing. More and more stacked GMOs have been developed for more improved functional properties and/or a stronger intended characteristic, such as antipest, improved product efficiency etc. Bt11 x GA21 is a new kind of stacked GM maize developed by Monsanto Company. Since there are no unique flanking sequences in stacked GMOs, up to now, no appropriate method has been reported to accurately detect them. In this passage, a novel universal primer multiplex PCR (UP-M-PCR) was developed and applied as a rapid screening method for the simultaneous detection of five target sequences (NOS, 35S, Bt11 event, GA21 event, and IVR) in maize Bt11 x GA21. This method overcame the disadvantages rooted deeply in conventional multiplex PCR such as complex manipulation, lower sensitivity, self-inhibition and amplification disparity resulting from different primers. What's more, it got a high specificity and had a detection limit of 0.1% (approximates to 38 haploid genome copies). Furthermore, real-time PCR combined with multivariate statistical analysis was used for accurate quantification of stacked GM maize Bt11 x GA21 in 100% GM maize mixture (Bt11 x GA21, Bt11 and GA21). Detection results showed that this method could accurately validate the content of Bt11, GA21 and Bt11 x GA21 in 100% GM mixture with a detection limit of 0.5% (approximates to 200 haploid genome copies) and a low relative standard deviation <5%. All the data proved that this method may be widely applied in event-specific detection of other stacked GMOs in GM-mixture. Publication Types: Evaluation Studies Research Support, Non-U.S. Gov't PMID: 19105640 [PubMed - indexed for MEDLINE] 14: Regul Toxicol Pharmacol. 2008 Dec 7. [Epub ahead of print] Murine models for evaluating the allergenicity of novel proteins and foods. Aldemir H, Bars R, Herouet-Guicheney C. University of Paris Sud XI, Faculty of Pharmacy, 5 rue J.B. Clément, 92290 Châtenay Malabry, France; Bayer CropScience, 355 rue Dostoïevski, 06903 Sophia-Antipolis, France. Genetically modified crops convey many benefits to world population. However, a rigorous safety assessment procedure, including an evaluation of the allergenic potential, is fundamental before their release into the food chain. As an integral part of the safety assessment process, regulatory authorities worldwide strongly recommend the use of tests that can predict the allergenic potential of the novel proteins. All guidance documents are based on an array of tests that have been proposed in 2003 by the Codex Alimentarius. Although the animal model is not a requirement of the Codex Alimentarius weight of evidence approach, allergenic hazard of novel proteins could only be evaluated by an in vivo model that can potentially identify and distinguish commonly allergenic proteins from rarely allergenic proteins. Therefore, food allergy experts encourage its development. During the 2007 International Life Science Institute (ILSI) workshop (Nice, France), worldwide experts shared their latest research results on rodent models to evaluate the allergenic potential of proteins and foods. This review presents the most promising rodent models for assessing food protein allergenicity that were evaluated during this ILSI workshop. PMID: 19100305 [PubMed - as supplied by publisher] 15: Plant Cell Rep. 2009 Mar;28(3):445-55. Epub 2008 Dec 18. Evaluation of a morphological marker selection and excision system to generate marker-free transgenic cassava plants. Saelim L, Phansiri S, Suksangpanomrung M, Netrphan S, Narangajavana J. Department of Biotechnology, Faculty of Science, Mahidol University, Rama 6 Road, Bangkok, 10400, Thailand. The efficacy of the ipt-type Multi-Auto-Transformation (MAT) vector system to transform the extensively grown cassava cultivar "KU50" was evaluated. This system utilizes the isopentenyltransferase (ipt) gene as morphological marker for visual selection of transgenic lines. The extreme shooty phenotype (ESP) of transgenic lines is lost due to the removal of ipt gene mediated by the yeast Rint/RS system. As a result, phenotypically normal shoots, considered marker-free transgenic plants, could be obtained. When transforming KU50 cassava cultivar with two different ipt-type MAT vectors, transformation frequency at 19-21% was observed. Among the total number of ESP explants, 32-38% regained normal extended shoot phenotype and 88-96% of which were confirmed to represent the marker-free transgenic plants. This is the first demonstration of the efficacy of Rint/RS system in promoting excision of ipt marker gene in cassava specie, with the consequent rapid production of marker-free transgenic plants. The high efficiency of this system should facilitate pyramiding a number of transgenes by repeated transformation without having to undergo through laborious, expensive and time-consuming processes of sexual crossing and seed production. The generation of marker-free, thus environmentally safe, genetically modified cassava clones should also ease the public concerns regarding the use of transgenic cassava in both food and nonfood industries. PMID: 19093119 [PubMed - in process] 16: Environ Biosafety Res. 2008 Oct-Dec;7(4):241-52. Epub 2008 Dec 16. Dispersal of viable row-crop seeds of commercial agriculture by farmland birds: implication for genetically modified crops. Cummings JL, Handley LW, Macbryde B, Tupper SK, Werner SJ, Byram ZJ. U.S. Department of Agriculture, Animal and Plant Health Inspection Service, Wildlife Services, National Wildlife Research Center, 4101 LaPorte Avenue, Fort Collins, CO 80521, USA. john.1.cummings@aphis.usda.gov To address some concerns about the expansion of genetically engineered pharmaceutical and industrial crops to outdoor plantings and potential impacts on the human food supply, we determined whether commercial agriculture seeds of maize or corn Zea mays L., barley Hordeum vulgare L., safflower Carthamus tinctorius L. and rice Oryza sativa L. are digested or pass viably through the digestive tract, or are transported externally, by captive mallard ducks Anas platyrhynchos L., ring-necked pheasants Phasianus colchicus L., red-winged blackbirds Agelaius phoeniceus (L.) and rock pigeons Columba livia Gmelin (with the exception of whole maize seeds which were too large to feed to the blackbirds). These crop seeds, whether free-fed or force-fed, did not pass through the digestive tract of these bird species. The birds nonetheless did retain viable seeds in the esophagus/crop and gizzard for several hours. For example, after foraging for 6 h, mallards had retained an average of 228 +/- 112 barley seeds and pheasants 192 +/- 78 in the esophagus/crop, and their germination rates were 93 and 50%, respectively. Birds externally transported seeds away from the feeding location, but in only four instances were seeds found attached to their muddy feet or legs and in no case to feathers. Risk of such crop seeds germinating, establishing and reproducing off site after transport by a bird (externally or internally) or movement of a carcass by a predator, will depend greatly on the crop and bird species, location, environmental conditions (including soil characteristics), timing, and seed condition. Publication Types: Research Support, U.S. Gov't, Non-P.H.S. PMID: 19081011 [PubMed - in process] 17: New Phytol. 2009;181(1):174-86. Effects of elevated carbon dioxide and ozone on volatile terpenoid emissions and multitrophic communication of transgenic insecticidal oilseed rape (Brassica napus). Himanen SJ, Nerg AM, Nissinen A, Pinto DM, Stewart CN Jr, Poppy GM, Holopainen JK. Department of Environmental Science, University of Kuopio, P.O. Box 1627, FIN-70211 Kuopio, Finland. sari.himanen@uku.fi Does transgenically incorporated insect resistance affect constitutive and herbivore-inducible terpenoid emissions and multitrophic communication under elevated atmospheric CO(2) or ozone (O(3))? This study aimed to clarify the possible interactions between allocation to direct defences (Bacillus thuringiensis (Bt) toxin production) and that to endogenous indirect defences under future climatic conditions. Terpenoid emissions were measured from vegetative-stage non-Bt and Bt Brassica napus grown in growth chambers under control or doubled CO(2), and control (filtered air) or 100 ppb O(3). The olfactometric orientation of Cotesia vestalis, an endoparasitoid of the herbivorous diamondback moth (Plutella xylostella), was assessed under the corresponding CO(2) and O(3) concentrations. The response of terpenoid emission to CO(2) or O(3) elevations was equivalent for Bt and non-Bt plants, but lower target herbivory reduced herbivore-inducible emissions from Bt plants. Elevated CO(2) increased emissions of most terpenoids, whereas O(3) reduced total terpenoid emissions. Cotesia vestalis orientated to host-damaged plants independent of plant type or CO(2) concentration. Under elevated O(3), host-damaged non-Bt plants attracted 75% of the parasitoids, but only 36.8% of parasitoids orientated to host-damaged Bt plants. Elevated O(3) has the potential to perturb specialized food-web communication in Bt crops. Publication Types: Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. PMID: 19076723 [PubMed - indexed for MEDLINE] 18: Food Chem Toxicol. 2009 Feb;47(2):425-32. Epub 2008 Dec 6. A 90-day toxicology study of transgenic lysine-rich maize grain (Y642) in Sprague-Dawley rats. He XY, Tang MZ, Luo YB, Li X, Cao SS, Yu JJ, Delaney B, Huang KL. College of Food Science and Nutritional Engineering, China Agricultural University, No. 17 Tsinghua Donglu, Beijing 100083, China. The gene for a lysine-rich protein (sb401) obtained from potatoes (Solanum berthaultii) was inserted into maize seed to produce Y642 transgenic maize. Compositional analysis of Y642 grain demonstrated that the concentrations of lysine and total protein were higher than those observed in maize grain from a near-isogenic non-genetically modified (non-GM) commercially available control quality protein maize (Nongda 108). The safety of Y642 maize grain was assessed by comparison of toxicology response variables in Sprague-Dawley (SD) rats consuming diets containing Y642 maize grain with those containing Nongda 108 maize grain. Maize grains from Y642 or Nongda 108 were incorporated into rodent diets at low (30%) or high concentrations (76%) and administered to SD rats (n=10/sex/group) for 90 days. An additional group of negative control group of rats (n=10/sex/group) were fed AIN93G diets. No adverse diet-related differences in body weights, feed consumption/utilization, clinical chemistry, hematology, absolute and relative organ weights were observed. Further, no differences in gross or microscopic pathology were observed between rats consuming diets with Y642 maize grain compared with rats consuming diets containing Nongda 108 maize grain. These results demonstrated that Y642 lysine-rich maize is as safe and nutritious as conventional quality protein maize. Publication Types: Comparative Study Research Support, Non-U.S. Gov't PMID: 19073230 [PubMed - indexed for MEDLINE] 19: J Agric Food Chem. 2009 Jan 14;57(1):26-37. Real-time PCR array as a universal platform for the detection of genetically modified crops and its application in identifying unapproved genetically modified crops in Japan. Mano J, Shigemitsu N, Futo S, Akiyama H, Teshima R, Hino A, Furui S, Kitta K. National Food Research Institute, 2-1-12 Kannondai, Tsukuba, Ibaraki, Japan. We developed a novel type of real-time polymerase chain reaction (PCR) array with TaqMan chemistry as a platform for the comprehensive and semiquantitative detection of genetically modified (GM) crops. Thirty primer-probe sets for the specific detection of GM lines, recombinant DNA (r-DNA) segments, endogenous reference genes, and donor organisms were synthesized, and a 96-well PCR plate was prepared with a different primer-probe in each well as the real-time PCR array. The specificity and sensitivity of the array were evaluated. A comparative analysis with the data and publicly available information on GM crops approved in Japan allowed us to assume the possibility of unapproved GM crop contamination. Furthermore, we designed a Microsoft Excel spreadsheet application, Unapproved GMO Checker version 2.01, which helps process all the data of real-time PCR arrays for the easy assumption of unapproved GM crop contamination. The spreadsheet is available free of charge at http://cse.naro.affrc.go.jp/jmano/index.html . Publication Types: Research Support, Non-U.S. Gov't PMID: 19072282 [PubMed - indexed for MEDLINE] 20: J Agromedicine. 2008;13(4):219-24. Biofuels and North American agriculture--implications for the health and safety of North American producers. Gunderson PD. Dakota Center for Technology-Optimized Agriculture, Devils Lake, ND 58301, USA. Paul.D.Gunderson.1@LRSC.NODAK.EDU This decade has provided North American agricultural producers with opportunity to not only produce fiber and food, but also fuel and other industrial products. The drivers incenting this development could be sustained well into the future, therefore workforce safety and health implications are likely to persist for some time. Within production agriculture, the 'feedstock growth and harvest cycle' and 'transport' sectors possess the changing exposures experienced by workers. The Conference explored the following exposures: distiller's grains and bio-processing byproducts, spent catalyst, solvent brine, microbial agents, genetically modified organisms, discharge effluent, H2O dilutes, change in cropping patterns and resultant use of different seeding and harvest technologies, pests (whether target or non-target), and rural traffic resulting from concentrated movement of massive quantities of biomass and grain. Other issues of a more general public health nature such as watershed implications, other environmental impacts, emissions, uneven economic development potential, public safety issues associated with transport of both fuel and other industrial products, and rural emergency medical service need were explored. And, agronomic impacts were noted, including tillage change, potassium buildup in soil, nutrient depletion, sedimentation and erosion of tillable soil, and local esthetics. It was concluded that rural venues for formation and exploration of public policy need to be created. PMID: 19064413 [PubMed - indexed for MEDLINE] 21: Biosci Biotechnol Biochem. 2008 Dec;72(12):3301-5. Epub 2008 Dec 7. Accumulation of the bioactive peptides, novokinin, LPYPR and rubiscolin, in seeds of genetically modified soybean. Nishizawa K, Kita A, Doi C, Yamada Y, Ohinata K, Yoshikawa M, Ishimoto M. National Agricultural Research Center for Hokkaido Region, Sapporo, Hokkaido, Japan. Novokinin (RPLKPW), LPYPR, and rubiscolin (YPLDLF) are bioactive peptides with respective hypotensive, hypocholesterolemic, and memory-enhancing activities. We generated transgenic soybean lines that expressed modified forms of the alpha' subunit of seed storage protein beta-conglycinin containing tandem repeats of these bioactive peptides. The modified alpha' subunits constituted up to 0.2% of extracted proteins from the transgenic seeds. Publication Types: Research Support, Non-U.S. Gov't PMID: 19060385 [PubMed - indexed for MEDLINE] 22: Maturitas. 2009 Jan 20;62(1):42-6. Epub 2008 Dec 5. Morphological modification of female bladder after prolonged use of soy-based diets. da Silva Faria T, Soares LL, Medeiros JL Jr, Boaventura GT, Sampaio FJ, da Fonte Ramos C. Urogenital Research Unit-UERJ, Biomedical Center, State University of Rio de Janeiro, Av. 28 de Setembro, 87-fundos-FCM-terreo, 20551-030 Rio de Janeiro, RJ, Brazil. OBJECTIVES: The aim of this study was to compare the effects of a prolonged use of organic and transgenic soy upon the lipid profile and the collagen/muscle ratio of the detrusor muscle of the bladder. METHODS: Wistar rats were fed three different diets from weaning until sacrifice (15 months old): control group (CG) casein-based diet; organic soy group (OSG) organic soy-based diet; genetically modified soy group (GMSG) transgenic soy-based diet. RESULTS: There was no difference in the food consumption or in the diet isoflavone components among the groups. Comparing to CG, both OSG and GMSG groups presented a significant (p<0.05) reduction in the body weight, triglycerides, cholesterol and the smooth muscle of the detrusor and a significant (p<0.05) increase of collagen fibers number of the detrusor muscle. CONCLUSIONS: These findings call into question that, the prolonged use of soy-based diets can be deleterious to the bladder by altering the collagen/muscle ratio what can cause bladder dysfunctions similar with that occurring during menopause. PMID: 19058935 [PubMed - in process] 23: BMC Genomics. 2008 Dec 4;9:584. Optimised padlock probe ligation and microarray detection of multiple (non-authorised) GMOs in a single reaction. Prins TW, van Dijk JP, Beenen HG, Van Hoef AA, Voorhuijzen MM, Schoen CD, Aarts HJ, Kok EJ. RIKILT - Institute of Food Safety (WUR), Bornsesteeg 45, 6708 PD Wageningen, the Netherlands. theo.prins@wur.nl BACKGROUND: To maintain EU GMO regulations, producers of new GM crop varieties need to supply an event-specific method for the new variety. As a result methods are nowadays available for EU-authorised genetically modified organisms (GMOs), but only to a limited extent for EU-non-authorised GMOs (NAGs). In the last decade the diversity of genetically modified (GM) ingredients in food and feed has increased significantly. As a result of this increase GMO laboratories currently need to apply many different methods to establish to potential presence of NAGs in raw materials and complex derived products. RESULTS: In this paper we present an innovative method for detecting (approved) GMOs as well as the potential presence of NAGs in complex DNA samples containing different crop species. An optimised protocol has been developed for padlock probe ligation in combination with microarray detection (PPLMD) that can easily be scaled up. Linear padlock probes targeted against GMO-events, -elements and -species have been developed that can hybridise to their genomic target DNA and are visualised using microarray hybridisation.In a tenplex PPLMD experiment, different genomic targets in Roundup-Ready soya, MON1445 cotton and Bt176 maize were detected down to at least 1%. In single experiments, the targets were detected down to 0.1%, i.e. comparable to standard qPCR. CONCLUSION: Compared to currently available methods this is a significant step forward towards multiplex detection in complex raw materials and derived products. It is shown that the PPLMD approach is suitable for large-scale detection of GMOs in real-life samples and provides the possibility to detect and/or identify NAGs that would otherwise remain undetected. Publication Types: Research Support, Non-U.S. Gov't PMID: 19055784 [PubMed - in process] 24: Plant Biotechnol J. 2009 Feb;7(2):119-28. Epub 2008 Nov 26. Recombinant aryl hydrocarbon receptors for bioassay of aryl hydrocarbon receptor ligands in transgenic tobacco plants. Kodama S, Okada K, Akimoto K, Inui H, Ohkawa H. Graduate School of Science and Technology, Kobe University, Rokkodai-cho 1-1, Nada-ku, Kobe 657-8501, Japan. Dioxin residues widely contaminate soil and agricultural products at low concentrations and may accumulate in organisms at the top of food chains owing to their physicochemical properties. In this study, we have developed novel, dioxin-inducible, reporter gene expression systems regulated by recombinant aryl hydrocarbon receptors (AhRs). The recombinant AhRs, referred to as XDVs, consist of the DNA-binding domain of the bacterial repressor protein LexA, a 90-kDa heat shock protein- and ligand-binding regulatory domain from mouse AhR, and the transactivation domain of herpes simplex virus regulatory protein VP16. Transgenic tobacco plants carrying XDVs absorb various AhR ligands, including 3-methylcholanthrene, beta-naphthoflavone and indigo from solid medium and vermiculite, and show dose- and time-dependent expression of the beta-glucuronidase reporter gene. The results clearly suggest that XDVs are functional transcription factors that respond to AhR ligands, and that the XDV-mediated reporter gene expression system is applicable to bioassays for dioxin residues in the environment. Publication Types: Research Support, Non-U.S. Gov't PMID: 19055610 [PubMed - indexed for MEDLINE] 25: Nature. 2008 Dec 4;456(7222):570. Comment on: Nature. 2008 Oct 16;455(7215):850-2. Switch to ecological engineering would aid independence. Settele J, Biesmeijer J, Bommarco R. Publication Types: Comment Letter PMID: 19052603 [PubMed - indexed for MEDLINE] 26: Nature. 2008 Dec 4;456(7222):563-8. Agronomy: Five crop researchers who could change the world. Marris E. Publication Types: News PMID: 19052600 [PubMed - indexed for MEDLINE] 27: Vopr Pitan. 2008;77(5):13-7. [Medical and biological safety assessment of genetically modified maize event MON 88017. Report 2. Genotoxicologic, immunologic and allergologic examinations] [Article in Russian] Tyshko NV, Britsina MV, Gmoshinskiĭ IV, Zhanataev AK, Zakharova NS, Zorin SN, Mazo VK, Semenov BF. There are presented the results of genotoxicologic, immunologic and allergologic examinations which were conducted within the framework of integrated medical and biological assessment of genetically modified rootworm Diabrotica spp.--protected and glyphosate tolerant maize event MON 88017. Analysis of damages of DNA and structural chromosome aberrations, assessment of the allergenic potential and immunoreactive properties has not confirmed any genotoxic, allergenic and immunotoxic effect of maize event MON 88017. Publication Types: English Abstract PMID: 19048882 [PubMed - indexed for MEDLINE] 28: Vopr Pitan. 2008;77(5):4-12. [Medical and biological safety assessment of genetically modified maize event MON 88017. Report 1. Toxicologo-hygienic examinations] [Article in Russian] Tutel'ian VA, Gapparov MM, Avren'eva LI, Aksiuk IN, Guseva GV, kravchenko LV, L'vova LS, Saprykin VP, Tyshko NV, Chernysheva ON. The results of toxicologo-hygienic examinations, which were conducted within the framework of integrated medical and biological assessment of genetically modified rootworm Diabrotica spp.--protected and glyphosate tolerant maize event MON 88017, are presented. Analysis of morphological, hematological, biochemical parameters and system (sensitive) biomarkers has not confirmed any toxic effect of maize event MON 88017. Publication Types: English Abstract PMID: 19048881 [PubMed - indexed for MEDLINE] 29: Environ Sci Pollut Res Int. 2009 Jan;16(1):85-94. Epub 2008 Dec 2. Cumulative impact of GM herbicide-tolerant cropping on arable plants assessed through species-based and functional taxonomies. Squire GR, Hawes C, Begg GS, Young MW. Scottish Crop Research Institute, Invergowrie, Dundee, DD2 5DA, UK, geoff.squire@scri.ac.uk. BACKGROUND, AIM AND SCOPE: In a gradualist approach to the introduction of crop biotechnology, the findings of experimentation at one scale are used to predict the outcome of moving to a higher scale of deployment. Movement through scales had occurred for certain genetically modified herbicide-tolerant (GMHT) crops in the UK as far as large-scale field trials. However, the land area occupied by these trials was still <1% of the area occupied by the respective non-GM crops. Some means is needed to predict the direction and size of the effect of increasing the area of GMHT cropping on ecological variables such as the diversity among species and trophic interactions. Species-accumulation curves are examined here as a method of indicating regional-scale impacts on botanical diversity from multiple field experiments. MATERIALS AND METHODS: Data were used from experiments on the effect of (GMHT) crops and non-GM, or conventional, comparators in fields sown with four crop types (beet, maize, spring and winter oilseed rape) at a total of 250 sites in the UK between 2000 and 2003. Indices of biodiversity were measured in a split-field design comparing GMHT with the farmers' usual weed management. In the original analyses based on the means at site level, effects were detected on the mass of weeds in the three spring crops and the proportion of broadleaf and grass weeds in winter oilseed rape, but not on indices of plant species diversity. To explore the links between site means and total taxa, accumulation curves were constructed based on the number of plant species (a pool of around 250 species in total) and the number of plant functional types (24), inferred from the general life-history characteristics of a species. RESULTS: Species accumulation differed between GMHT and conventional treatments in direction and size, depending on the type of crop and its conventional management. Differences were mostly in the asymptote of the curve, indicative of the maximum number of species found in a treatment, rather than the steepness of the curve. In winter oilseed rape, 8% more species were accumulated in the GMHT treatment, mainly as a result of the encouragement of grass species by the herbicide when applied in the autumn. (Overall, GMHT winter oilseed rape had strong negative effects on both the food web and the potential weed burden by increasing the biomass of grasses and decreasing that of broadleaf weeds.) In maize, 33% more species-a substantial increase-were accumulated in the GMHT than in the conventional, consistent with the latter's highly suppressive weed management using triazine herbicides. In the spring oilseed rape and beet, fewer species (around 10%) were accumulated in the GMHT than the conventional. The GMHT treatments did not remove or add any functional (life history) types, however. Differences in species accumulation between treatments appeared to be caused by loss or gain of rarer species. The generality of this effect was confirmed by simulations of species accumulation in which the species complement at each of 50 sites was drawn from a regional pool and subjected to reducing treatment at each site. Shifts in the species-accumulation parameters, comparable to those measured, occurred only when a treatment removed the rarer species at each site. DISCUSSION: Species accumulation provided a set of simple curve-parameters that captured the net result of numerous local effects of treatments on plant species and, in some instances, the balance between grass and broadleaf types. The direction of effect was not the same in the four crops and depended on the severity of the conventional treatment and on complex interactions between season, herbicide and crop. The accumulation curves gave an indication of potential positive or negative consequences for regional species pools of replacing a conventional practice with GMHT weed management. In this and related studies, a range of indicators, through which diversity was assessed by both species and functional type, and at both site and regional scales, gave more insight into effects of GMHT treatment than provided by any one indicator. CONCLUSIONS: Species accumulation was shown to discriminate at the regional scale between agronomic treatments that had little effect on species number at the field scale. While a comprehensive assessment of GM cropping needs to include an examination of regional effects, as here, the costs of doing this in all instances would be prohibitive. Simulations of diversity-reducing treatments could provide a theoretical framework for predicting the likely regional effects from in-field plant dynamics. RECOMMENDATIONS AND PERSPECTIVES: Accumulation curves potentially offer a means of linking within-site effects to regional impacts on biodiversity resulting from any change in agricultural practice. To guide empirical measurement, there is a scope to apply a methodology such as individual-based modelling at the field scale to explore the links between agronomic treatments and the relative abundance of plant types. The framework needs to be validated in practice, using species-based and functional taxonomies, the latter defined by measured rather than inferred traits. PMID: 19048321 [PubMed - in process] 30: Nature. 2008 Nov 27;456(7221):421-2. A fruitless campaign. [No authors listed] Publication Types: Editorial PMID: 19037266 [PubMed - indexed for MEDLINE] 31: J Agric Food Chem. 2008 Dec 24;56(24):12099-104. Safety assessment of bacterial choline oxidase protein introduced in transgenic crops for tolerance against abiotic stress. Singh AK, Singh BP, Prasad GB, Gaur SN, Arora N. Institute of Genomics and Integrative Biology (CSIR), Delhi, India. Genetically modified crops have resistance to abiotic stress by introduction of choline oxidase protein. In the present study, the safety of choline oxidase protein derived from Arthrobacter globiformis was assessed for toxicity and allergenicity. The protein was stable at 90 degrees C for 1 h. Toxicity studies of choline oxidase in mice showed no significant difference (p > 0.05) from control in terms of growth, body weight, food consumption, and blood biochemical indices. Histology of gut tissue of mice fed protein showed normal gastric mucosal lining and villi in jejunum and ileum sections. Specific IgE in serum and IL-4 release in splenic culture supernatant were low in choline oxidase treated mice, comparable to control. Intravenous challenge with choline oxidase did not induce any adverse reaction, unlike ovalbumin group mice. Histology of lung tissues from choline oxidase sensitized mice showed normal airways, whereas ovalbumin-sensitized mice showed inflamed airways with eosinophilic infiltration and bronchoconstriction. ELISA carried out with food allergic patients' sera revealed no significant IgE affinity with choline oxidase. Also, choline oxidase did not show any symptoms of toxicity and allergenicity in mice. Publication Types: Research Support, Non-U.S. Gov't PMID: 19035641 [PubMed - indexed for MEDLINE] 32: Regul Toxicol Pharmacol. 2008 Nov 8. [Epub ahead of print] Identifying food proteins with allergenic potential: Evolution of approaches to safety assessment and research to provide additional tools. Ladics GS, Selgrade MK. DuPont Co., DuPont Crop Genetics, Wilmington, DE 19880 USA. A safety assessment process exists for genetically engineered crops that includes the evaluation of the expressed protein for allergenic potential. The objectives of this evaluation are twofold: (1) to protect allergic consumers from exposure to known allergenic or cross-reactive proteins, and (2) protect the general population from risks associated with the introduction of genes encoding proteins that are likely to become food allergens. The first systematic approach to address these concerns was formulated by Metcalfe et al. [Metcalfe, D.D., Astwood, J.D., Townsend, R., Sampson, H.A., Taylor, S.L., and Fuchs, R.L. 1996. Assessment of the allergenic potential of foods from genetically engineered crop plants. Crit. Rev. Food Sci. Nutr. 36(5), 165-186.] and subsequently Food and Agriculture Organization of the United Nations/World Health Organization (FAO/WHO) [FAO/WHO, 2001. Evaluation of allergenicity of genetically modified foods. Report of a Joint FAO/WHO Expert Consultation on Allergenicity of Foods Derived from Biotechnology. January 22-25, 2001. Rome, Italy]. More recently, Codex [Codex Alimentarius Commission, 2003. Alinorm 03/34: Joint FAO/WHO Food Standard Programme, Codex Alimentarius Commission, Twenty-Fifth Session, Rome, Italy, 30 June-5 July, 2003. Appendix III, Guideline for the conduct of food safety assessment of foods derived from recombinant-DNA plants, and Appendix IV, Annex on the assessment of possible allergenicity. pp. 47-60], noting that no single factor is recognized as an identifier for protein allergenicity, suggested a weight of evidence approach be conducted that takes into account a variety of factors and approaches for an overall assessment of allergenic potential. These various recommendations are based on what is known about allergens, including the history of exposure and safety of the gene(s) source; amino acid sequence identity to human allergens; stability to pepsin digestion in vitro; protein abundance in the crop and processing effects; and when appropriate, specific IgE binding studies or skin-prick testing. Similarities and differences between these various suggested recommendations, as well as data gaps, are discussed. The US Environmental Protection Agency (EPA)'s Office of Research and Development (ORD) has initiated a targeted research effort to address data gaps and improve the various recommended methods/endpoints for assessing the allergenic risks associated with plant incorporated pesticides (PIPs) through both intramural and extramural (grant supported) research. The areas of primary focus for EPA include: (1) development and evaluation of animal models; (2) targeted or specific serological assays; and (3) structure-activity relationships. Details on the current as well as proposed EPA funded research are discussed. More recently US EPA has partnered with the National Institute of Allergy and Infectious Disease (NIAID), National Institutes of Health to support research in areas of mutual interest with respect to food allergy. PMID: 19028539 [PubMed - as supplied by publisher] 33: Plant Biotechnol J. 2009 Jan;7(1):106-17. Epub 2008 Oct 13. Sensory analysis of calcium-biofortified lettuce. Park S, Elless MP, Park J, Jenkins A, Lim W, Chambers E 4th, Hirschi KD. Department of Horticulture, Forestry and Recreation Resources, Kansas State University, Manhattan, KS 66506, USA. shpark@ksu.edu Vegetables represent an attractive means of providing increased calcium nutrition to the public. In this study, it was demonstrated that lettuce expressing the deregulated Arabidopsis H(+)/Ca(2+) transporter sCAX1 (cation exchanger 1) contained 25%-32% more calcium than controls. These biofortified lettuce lines were fertile and demonstrated robust growth in glasshouse growth conditions. Using a panel of highly trained descriptive panellists, biofortified lettuce plants were evaluated and no significant differences were detected in flavour, bitterness or crispness when compared with controls. Sensory analysis studies are critical if claims are to be made regarding the efficacy of biofortified foods, and may be an important component in the public acceptance of genetically modified foods. Publication Types: Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. PMID: 19021875 [PubMed - indexed for MEDLINE] 34: J Biol. 2008 Nov 7;7(9):33. [Epub ahead of print] Aquaglyceroporins: ancient channels for metalloids. Bhattacharjee H, Mukhopadhyay R, Thiyagarajan S, Rosen BP. Department of Biochemistry and Molecular Biology, Wayne State University, School of Medicine, Detroit, MI 48201, USA. brosen@med.wayne.edu. ABSTRACT: The identification of aquaglyceroporins as uptake channels for arsenic and antimony shows how these toxic elements can enter the food chain, and suggests that food plants could be genetically modified to exclude arsenic while still accumulating boron and silicon. PMID: 19014407 [PubMed - as supplied by publisher] 35: Plant Foods Hum Nutr. 2009 Mar;64(1):1-5. Organic and genetically modified soybean diets: consequences in growth and in hematological indicators of aged rats. Daleprane JB, Feijó TS, Boaventura GT. College of Medicine, Federal Fluminense University, Niterói, Brazil, juliobd@gmail.com. The aim of this study was to evaluate the protein quality of organic and genetically modified soy by feeding specific diets to rats. Three groups of Wistar rats (n = 10) were used, and each group was named according to the food that they ate. There was an organic soy group (OG), a genetically modified soy group (GG), and a control group (CG). All animals received water and diet ad libitum for 455 days. At the end of this period, the weight of the GG group was the same as that of the OG, and both were higher than CG. Protein intake was similar for the OG and GG, which were significantly lower (p < 0.0005) than the CG. The growth rate (GR) of the rats, albumin levels, and total levels of serum protein were comparable for all groups. Hematocrit (p < 0.04) and hemoglobin (p < 0.03) for the OG and GG were less than the CG. Although the OG and GG demonstrated reduced hematocrit and hemoglobin, both types of soy were utilized in a way similar to casein. This result suggests that the protein quality of soy is parallel to the standard protein casein in terms of growth promotion but not hematological indicators. PMID: 19011971 [PubMed - in process] 36: Methods Mol Biol. 2009;478:305-14. Design and management of field trials of transgenic cereals. Bedo Z, Rakszegi M, Láng L. Agricultural Research Institute of the Hungarian Academy of Sciences, Hungary. The development of gene transformation systems has allowed the introgression of alien genes into plant genomes, thus providing a mechanism for broadening the genetic resources available to plant breeders. The design and the management of field trials vary according to the purpose for which transgenic cereals are developed. Breeders study the phenotypic and genotypic stability of transgenic plants, monitor the increase in homozygosity of transgenic genotypes under field conditions, and develop backcross generations to transfer the introduced genes into secondary transgenic cereal genotypes. For practical purposes, they may also multiply seed of the transgenic lines to produce sufficient amounts of grain for the detailed analysis of trait(s) of interest, to determine the field performance of transgenic lines, and to compare them with the non-transformed parental genotypes. Prior to variety registration, the Distinctness, Uniformity and Stability (DUS) tests and Value for Cultivation and Use (VCU) experiments are carried out in field trials. Field testing includes specific requirements for transgenic cereals to assess potential environmental risks. The capacity of the pollen to survive, establish and disseminate in the field test environment, the potential for gene transfer, the effects of products expressed by the introduced sequences and phenotypic and genotypic instability that might cause deleterious effects must all be specifically monitored, as required by EU Directives 2003/701/EC (1) on the release of genetically modified higher plants in the environment. PMID: 19009453 [PubMed - indexed for MEDLINE] 37: Methods Mol Biol. 2009;478:273-88. Establishing substantial equivalence: proteomics. Lovegrove A, Salt L, Shewry PR. Department of Plant Sciences, Rothamsted Research, Centre for Crop Genetic Improvement, Harpenden, Hertfordshire, UK. Wheat is a major crop in world agriculture and is consumed after processing into a range of food products. It is therefore of great importance to determine the consequences (intended and unintended) of transgenesis in wheat and whether genetically modified lines are substantially equivalent to those produced by conventional plant breeding. Proteomic analysis is one of several approaches which can be used to address these questions. Two-dimensional PAGE (2D PAGE) remains the most widely available method for proteomic analysis, but is notoriously difficult to reproduce between laboratories. We therefore describe methods which have been developed as standard operating procedures in our laboratory to ensure the reproducibility of proteomic analyses of wheat using 2D PAGE analysis of grain proteins. Publication Types: Research Support, Non-U.S. Gov't PMID: 19009451 [PubMed - indexed for MEDLINE] 38: J Biosci Bioeng. 2008 Oct;106(4):350-6. Chemical characteristics and volatile profile of genetically modified peanut cultivars. Ng EC, Dunford NT, Chenault K. Department of Biosystems and Agricultural Engineering and Robert M. Kerr Food & Agricultural Products Center, Oklahoma State University, FAPC Room 103, Stillwater, OK 74078, USA. Genetic engineering has been used to modify peanut cultivars for improving agronomic performance and pest resistance. Food products developed through genetic engineering have to be assessed for their safety before approval for human consumption. Preservation of desirable chemical, flavor and aroma attributes of the peanut cultivars during the genetic modifications is critical for acceptance of genetically modified peanuts (GMP) by the food industry. Hence, the main objective of this study is to examine chemical characteristics and volatile profile of GMP. The genetically modified peanut cultivars, 188, 540 and 654 were obtained from the USDA-ARS in Stillwater, Oklahoma. The peanut variety Okrun was examined as a control. The volatile analysis was performed using a gas chromatograph/mass spectrometer (GC/MS) equipped with an olfactory detector. The peanut samples were also analyzed for their moisture, ash, protein, sugar and oil compositions. Experimental results showed that the variations in nutritional composition of peanut lines examined in this study were within the values reported for existing cultivars. There were minor differences in volatile profile among the samples. The implication of this study is significant, since it shows that peanut cultivars with greater pest and fungal resistance were successfully developed without major changes in their chemical characteristics. PMID: 19000610 [PubMed - in process] 39: Nat Biotechnol. 2008 Nov;26(11):1223-5. Coexistence in the EU-return of the moratorium on GM crops? Devos Y, Demont M, Sanvido O. Publication Types: Letter PMID: 18997757 [PubMed - indexed for MEDLINE] 40: Nat Biotechnol. 2008 Nov;26(11):1222-3. Pharming in crop commodities. Stewart CN Jr. Publication Types: Letter PMID: 18997756 [PubMed - indexed for MEDLINE] 41: Nat Biotechnol. 2008 Nov;26(11):1205-7. FDA transgenic animal guidance finally surfaces. Fox JL. Publication Types: News PMID: 18997745 [PubMed - indexed for MEDLINE] 42: Biosci Biotechnol Biochem. 2008 Nov;72(11):2953-8. Epub 2008 Nov 7. Real-time PCR method using capturing oligo-immobilized PCR tubes to determine the specific gene for soybean and genetically modified soybean in food matrices. Harikai N, Saito S, Abe M, Kondo K, Kitta K, Akiyama H, Teshima R, Kinoshita K. School of Pharmaceutical Sciences, Mukogawa Women's University, Hyogo, Japan. A new real-time PCR method using capturing oligo-immobilized PCR tubes is described. This method was used to detect specific genes for soybean and genetically modified (GM) soybean in food matrices. In a standard reaction using soybean genomic DNA and a capturing oligo for the lectin gene (Le1) immobilized on the tube, we examined the effects of such hybridization conditions as the location, length, and amount of the capturing oligo, and the incubation time and temperature. Under optimized conditions, the copy number of Le1 was determined in a concentration-dependent manner from soybean genomic DNA and soybean lysate (DNA 10-1000 ng, r=0.99; lysate 1-100%, r=0.99). The copy number of a Roundup Ready soybean (RRS) gene was also successfully detected in a concentration-dependent manner (1-100%, r=0.99) from GM soybean lysate, using PCR tubes with an immobilized capturing oligo for the transgene. Our data indicate that this is a rapid and simple method to determine specific genes for soybean and GM soybean in food matrices. Publication Types: Research Support, Non-U.S. Gov't PMID: 18997399 [PubMed - indexed for MEDLINE] 43: Crit Rev Food Sci Nutr. 2009 Feb;49(2):164-75. Health risks of genetically modified foods. Dona A, Arvanitoyannis IS. Department of Forensic Medicine and Toxicology, University of Athens, Medical School, Athens, Greece. As genetically modified (GM) foods are starting to intrude in our diet concerns have been expressed regarding GM food safety. These concerns as well as the limitations of the procedures followed in the evaluation of their safety are presented. Animal toxicity studies with certain GM foods have shown that they may toxically affect several organs and systems. The review of these studies should not be conducted separately for each GM food, but according to the effects exerted on certain organs it may help us create a better picture of the possible health effects on human beings. The results of most studies with GM foods indicate that they may cause some common toxic effects such as hepatic, pancreatic, renal, or reproductive effects and may alter the hematological, biochemical, and immunologic parameters. However, many years of research with animals and clinical trials are required for this assessment. The use of recombinant GH or its expression in animals should be re-examined since it has been shown that it increases IGF-1 which may promote cancer. PMID: 18989835 [PubMed - indexed for MEDLINE] 44: Nature. 2008 Nov 6;456(7218):2. Animals aren't drugs. [No authors listed] Publication Types: Editorial PMID: 18987684 [PubMed - indexed for MEDLINE] 45: J Am Vet Med Assoc. 2008 Nov 1;233(9):1370-3. Genetically engineered animals in the food supply. Burns K. Publication Types: News PMID: 18980485 [PubMed - indexed for MEDLINE] 46: Plant Foods Hum Nutr. 2008 Dec;63(4):163-9. Antidiabetic and antioxidant effects of polyphenols in brown alga Ecklonia stolonifera in genetically diabetic KK-A(y) mice. Iwai K. Department of Nutrition, Faculty of Health Sciences, Aomori University of Health and Welfare, Hamadate, Aomori, Japan. k_iwai@auhw.ac.jp The dietary intake and control of blood glucose levels are very important in hyperglycemic patients and alpha-glucosidase inhibitors are a cost-effective means to preventing the progression of diabetes. In search of a natural inhibitor from food materials, alpha-glucosidase inhibitory activity and the anti-hyperglycemic effects of a brown alga, Ecklonia stolonifera, were investigated using non-insulin dependent diabetic mice. Methanolic extract of E. stolonifera (MEE), which contains a high content of polyphenols, showed strong inhibition of alpha-glucosidase in vitro. Male KK-A(y) mice, a genetically non-insulin dependent diabetic model, showed hyperglycemia with aging, but the ingestion of MEE suppressed the increase in plasma glucose and lipid peroxidation levels in unfasted KK-A(y) mice dose dependently. In KK-A(y) mice, which were fed the MEE diet for 4 weeks, MEE moderated the elevation of plasma glucose levels after the oral administration of maltose. The polyphenols in MEE were estimated to be phlorotannins by HPLC-PDA and LC/MS analyses. These results demonstrate that E. stolonifera, seaweed typically used as a health food, has strong antidiabetic and antioxidant effects in vivo, thus, it may have beneficial properties in the prevention of diabetes and could be useful in the development of an antidiabetic pharmaceutical and functional food. PMID: 18958624 [PubMed - indexed for MEDLINE] 47: J Am Diet Assoc. 2008 Nov;108(11):1888-95. Nutritional genomics, polyphenols, diets, and their impact on dietetics. Barnes S. Department of Pharmacology and Toxicology, 452 McCallum Research Building, University of Alabama at Birmingham, 1918 University Blvd, Birmingham, AL 35294, USA. sbarnes@uab.edu Nutritional genomics offers a way to optimize human health and the quality of life. It is an attractive endeavor, but one with substantial challenges. It encompasses almost all known aspects of science, ranging from the genomes of humans, plants, and microorganisms, to the highest levels of food science, analytical science, computing, and statistics of large systems, as well as human behavior. This paper describes the underlying biochemistry that is targeted by the principal issues in nutritional genomics, which entails genomics, transcriptomics, proteomics, and metabolomics. A major feature relevant to nutritional genomics is the single nucleotide polymorphisms in genes that interact with nutrients and other bioactive food components. These genetic changes may lead to alterations in absorption, metabolism, and functional responses to bioactive nutritional factors. Bioactive food components may also regulate gene expression at the transcriptome, protein abundance, and/or protein turnover levels. Even if all of these variables are known, additional variables to be considered include the nutritional variability of the food (unprocessed and processed), the amount that is actually eaten, and the eating-related behaviors of those consuming the food. These challenges are explored within the context of soy intake. Finally, the importance of international cooperation in nutritional genomics research is presented. Publication Types: Research Support, N.I.H., Extramural Review PMID: 18954579 [PubMed - indexed for MEDLINE] 48: J Exp Bot. 2008;59(15):4075-82. Epub 2008 Oct 24. Involvement of the ethylene response pathway in dormancy induction in chrysanthemum. Sumitomo K, Narumi T, Satoh S, Hisamatsu T. National Institute of Floricultural Science (NIFS), National Agriculture and Food Research Organization (NARO), 2-1 Fujimoto, Tsukuba, Ibaraki 305-8519, Japan. Temperature plays a significant role in the annual cycling between growth and dormancy of the herbaceous perennial chrysanthemum (Chrysanthemum morifolium Ramat.). After exposure to high summer temperatures, cool temperature triggers dormancy. The cessation of flowering and rosette formation by the cessation of elongation are characteristic of dormant plants, and can be stimulated by exogenous ethylene. Thus, the ethylene response pathway may be involved in temperature-induced dormancy of chrysanthemum. Transgenic chrysanthemums expressing a mutated ethylene receptor gene were used to assess this involvement. The transgenic lines showed reduced ethylene sensitivity: ethylene causes leaf yellowing in wild-type chrysanthemums, but leaves remained green in the transgenic lines. Extension growth and flowering of wild-type and transgenic lines varied between temperatures: at 20 degrees C, the transgenic lines showed the same stem elongation and flowering as the wild type; at cooler temperatures, the wild type formed rosettes with an inability to flower and entered dormancy, but some transgenic lines continued to elongate and flower. This supports the involvement of the ethylene response pathway in the temperature-induced dormancy of chrysanthemum. At the highest dosage of ethephon, an ethylene-releasing agent, wild-type plants formed rosettes with an inability to flower and became dormant, but one transgenic line did not. This confirms that dormancy is induced via the ethylene response pathway. Publication Types: Research Support, Non-U.S. Gov't PMID: 18952907 [PubMed - indexed for MEDLINE] 49: Food Chem Toxicol. 2008 Dec;46(12):3808-17. Epub 2008 Oct 8. Subchronic feeding study of high oleic acid soybeans (Event DP-3Ø5423-1) in Sprague-Dawley rats. Delaney B, Appenzeller LM, Munley SM, Hoban D, Sykes GP, Malley LA, Sanders C. Pioneer, A DuPont Company, Pioneer Hi-Bred International Inc., Johnston, IA 50131-0552, USA. bryan.delaney@pioneer.com DP-3Ø5423-1 (305423) is a genetically-modified (GM) soybean that was produced by biolistic insertion of a gm-fad2-1 gene fragment and the gm-hra gene into the germline of soybean seeds. The gm-fad2-1 gene fragment cosuppresses expression of the endogenous FAD2-1 gene encoding the seed-specific omega-6 fatty acid desaturase resulting in higher concentrations of oleic acid (18:1) relative to linoleic acid (18:2). The gm-hra gene encoding a modified acetolactate synthase (ALS) enzyme was used as a selectable marker. In the current study, processed fractions (meal, hulls, and oil) from 305423 soybeans, non-GM soybeans with a similar genetic background (near isoline control) and three commercially-available non-GM varieties were used to formulate diets that were nutritionally comparable to PMI Certified Rodent LabDiet 5002. Diets were fed to young adult Crl:CD(SD) rats (12/sex/group) for approximately 90 days. Compared with rats fed the non-GM control diet, no biologically relevant differences were observed in rats fed the 305423 diet with respect to body weight/gain, food consumption/efficiency, mortality, clinical signs of toxicity, or ophthalmological observations. No test diet-related effects were observed on neurobehavioral assessments, organ weights, or clinical or anatomic pathology. These results demonstrated that 305423 soybeans are as safe and wholesome as non-GM soybeans. Publication Types: Research Support, Non-U.S. Gov't PMID: 18952136 [PubMed - indexed for MEDLINE] 50: Prim Care. 2008 Dec;35(4):769-88. Common foods and farming methods thought to promote health: what the data show. Chahbazi J, Grow S. McLaren Family Medicine Residency, G-3245 Beecher Road, Flint, MI 48532, USA. chahbazi@umich.edu What are the best dietary practices and farming methods to promote health? The answer may depend on whether one looks at the health of individuals or the health of the planet (planetary health or PH). PH will equate to a healthy ecosphere fostered by dietary/farming practices that are less resource-intense, potentially decreasing starvation and carbon emissions. Best practices also may depend on whether by health one means lack of observable disease (such as obesity, nutritional deficiency, diabetes, or cancer), optimal health (also known as wellness), or longevity. This article attempts to give an overview of the evidence as regards all of these aspects and definitions of health. PMID: 18928829 [PubMed - indexed for MEDLINE] 51: PLoS Genet. 2008 Oct;4(10):e1000222. Epub 2008 Oct 17. The HLH-6 transcription factor regulates C. elegans pharyngeal gland development and function. Smit RB, Schnabel R, Gaudet J. Genes and Development Research Group, Department of Biochemistry and Molecular Biology, University of Calgary, Calgary, Alberta, Canada. The Caenorhabditis elegans pharynx (or foregut) functions as a pump that draws in food (bacteria) from the environment. While the "organ identity factor" PHA-4 is critical for formation of the C. elegans pharynx as a whole, little is known about the specification of distinct cell types within the pharynx. Here, we use a combination of bioinformatics, molecular biology, and genetics to identify a helix-loop-helix transcription factor (HLH-6) as a critical regulator of pharyngeal gland development. HLH-6 is required for expression of a number of gland-specific genes, acting through a discrete cis-regulatory element named PGM1 (Pharyngeal Gland Motif 1). hlh-6 mutants exhibit a frequent loss of a subset of glands, while the remaining glands have impaired activity, indicating a role for hlh-6 in both gland development and function. Interestingly, hlh-6 mutants are also feeding defective, ascribing a biological function for the glands. Pharyngeal pumping in hlh-6 mutants is normal, but hlh-6 mutants lack expression of a class of mucin-related proteins that are normally secreted by pharyngeal glands and line the pharyngeal cuticle. An interesting possibility is that one function of pharyngeal glands is to secrete a pharyngeal lining that ensures efficient transport of food along the pharyngeal lumen. Publication Types: Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't PMID: 18927627 [PubMed - indexed for MEDLINE] 52: Science. 2008 Oct 17;322(5900):362-4. Q&A: China's scientist premier. Interview by Bruce Alberts. Jiabao W. Publication Types: Interview News PMID: 18927368 [PubMed - indexed for MEDLINE] 53: J Nutr Biochem. 2008 Oct 14. [Epub ahead of print] Use of a novel genetic mouse model to investigate the role of folate in colitis-associated colon cancer. Chapkin RS, Kamen BA, Callaway ES, Davidson LA, George NI, Wang N, Lupton JR, Finnell RH. Faculty of Nutrition, Texas A&M University, College Station, TX 77843-2253, USA; Department of Nutrition and Food Science, Texas A&M University, College Station, TX 77843-2253, USA; Center for Environmental and Rural Health, Texas A&M University, College Station, TX 77843-2253, USA; Vegetable and Fruit Improvement Center, Texas A&M University, College Station, TX 77843-2119, USA. Inflammatory bowel disease (IBD) patients are at high risk for developing folate deficiency and colon cancer. Since it is difficult to study the subtle global and gene-specific epigenetic mechanisms involved in folate-mediated tumor initiation and promotion, we have generated genetically modified mouse models by targeting the reduced folate carrier (RFC1) and folate-binding protein (Folbp1) genes. The transgenic mice were fed semi-purified diets for 8 weeks containing either normal (2 mg) or deficient (0.1 mg folate/kg diet) levels of folate. Compound heterozygous mice (Folbp1(+/-); RFC1(+/-)) fed an adequate folate diet exhibited a reduction in plasma folate concentrations compared to heterozygous (Folbp1(+/-)) and littermate wild-type mice (P<.05). In contrast, no differences were observed in colonic mucosa. Consumption of a low folate diet significantly reduced (three- to fourfold) plasma and tissue folate levels in all animal models, although plasma homocysteine levels were not altered. In order to elucidate the relationship between folate status and inflammation-associated colon cancer, animals were injected with azoxymethane followed by dextran sodium sulphate treatment in the drinking water. Mice were fed a normal folate diet and were terminated 5 weeks after carcinogen injection. The number of high multiplicity aberrant crypt foci per centimeter of colon was significantly elevated (P<.05) in compound Folbp1(+/-); RFC1(+/-) (3.5+/-0.4) mice as compared to Folbp1(+/-) (1.9+/-0.3) and wild-type control mice (1.1+/-0.1). These data demonstrate that the ablation of two receptor/carrier-mediated pathways for folate transport increases the risk for developing inflammation-associated colon cancer. PMID: 18926688 [PubMed - as supplied by publisher] 54: C R Biol. 2008 Oct;331(10):763-71. Epub 2008 Sep 4. Genetic and molecular approaches to improve nutritional value of Brassica napus L. seed. Nesi N, Delourme R, Brégeon M, Falentin C, Renard M. INRA-Agrocampus Rennes-University of Rennes1 Joint Laboratory, UMR118, Plant Genetics and Biotechnologies, BP 35327, 35653 Le Rheu cedex, France. nathalie.nesi@rennes.inra.fr Oilseed rape (Brassica napus L.) is a major oil crop that also supplies proteins for the feed industry. In order to reduce total cost production, the objective is to increase oil yield while reducing crop inputs (especially nitrogen and pesticides). Concomitantly, it is necessary to anticipate specific uses (e.g., fatty acid composition) and to ensure the valorisation of the by-products (rapeseed meal). By the past, improvement of seed quality focused on fatty acid balance and low seed glucosinolate content. Current goals include the breeding of yellow-seeded rapeseed lines with high content of seed oil. The use of molecular tools and the exploitation of Arabidopsis knowledge will be presented and discussed. Publication Types: Review PMID: 18926490 [PubMed - indexed for MEDLINE] 55: Nature. 2008 Oct 16;455(7215):850-2. Comment in: Nature. 2008 Dec 4;456(7222):570. Is China ready for GM rice? Qiu J. Publication Types: News PMID: 18923484 [PubMed - indexed for MEDLINE] 56: Electrophoresis. 2008 Sep;29(18):3801-9. Using multiple PCR and CE with chemiluminescence detection for simultaneous qualitative and quantitative analysis of genetically modified organism. Guo L, Qiu B, Chi Y, Chen G. Ministry of Education Key Laboratory of Analysis and Detection Technology for Food Safety, Fuzhou University, Fuzhou, P. R. China. In this paper, an ultrasensitive CE-CL detection system coupled with a novel double-on-column coaxial flow detection interface was developed for the detection of PCR products. A reliable procedure based on this system had been demonstrated for qualitative and quantitative analysis of genetically modified organism-the detection of Roundup Ready Soy (RRS) samples was presented as an example. The promoter, terminator, function and two reference genes of RRS were amplified with multiplex PCR simultaneously. After that, the multiplex PCR products were labeled with acridinium ester at the 5'-terminal through an amino modification and then analyzed by the proposed CE-CL system. Reproducibility of analysis times and peak heights for the CE-CL analysis were determined to be better than 0.91 and 3.07% (RSD, n=15), respectively, for three consecutive days. It was shown that this method could accurately and qualitatively detect RRS standards and the simulative samples. The evaluation in terms of quantitative analysis of RRS provided by this new method was confirmed by comparing our assay results with those of the standard real-time quantitative PCR (RT-QPCR) using SYBR Green I dyes. The results showed a good coherence between the two methods. This approach demonstrated the possibility for accurate qualitative and quantitative detection of GM plants in a single run. Publication Types: Evaluation Studies Research Support, Non-U.S. Gov't PMID: 18850650 [PubMed - indexed for MEDLINE] 57: PLoS Genet. 2008 Oct;4(10):e1000213. Epub 2008 Oct 10. A TRPV channel modulates C. elegans neurosecretion, larval starvation survival, and adult lifespan. Lee BH, Ashrafi K. Department of Physiology, University of California San Francisco, San Francisco, CA, USA. For most organisms, food is only intermittently available; therefore, molecular mechanisms that couple sensation of nutrient availability to growth and development are critical for survival. These mechanisms, however, remain poorly defined. In the absence of nutrients, newly hatched first larval (L1) stage Caenorhabditis elegans halt development and survive in this state for several weeks. We isolated mutations in unc-31, encoding a calcium-activated regulator of neural dense-core vesicle release, which conferred enhanced starvation survival. This extended survival was reminiscent of that seen in daf-2 insulin-signaling deficient mutants and was ultimately dependent on daf-16, which encodes a FOXO transcription factor whose activity is inhibited by insulin signaling. While insulin signaling modulates metabolism, adult lifespan, and dauer formation, insulin-independent mechanisms that also regulate these processes did not promote starvation survival, indicating that regulation of starvation survival is a distinct program. Cell-specific rescue experiments identified a small subset of primary sensory neurons where unc-31 reconstitution modulated starvation survival, suggesting that these neurons mediate perception of food availability. We found that OCR-2, a transient receptor potential vanilloid (TRPV) channel that localizes to the cilia of this subset of neurons, regulates peptide-hormone secretion and L1 starvation survival. Moreover, inactivation of ocr-2 caused a significant extension in adult lifespan. These findings indicate that TRPV channels, which mediate sensation of diverse noxious, thermal, osmotic, and mechanical stimuli, couple nutrient availability to larval starvation survival and adult lifespan through modulation of neural dense-core vesicle secretion. Publication Types: Research Support, Non-U.S. Gov't PMID: 18846209 [PubMed - indexed for MEDLINE] 58: Nat Biotechnol. 2008 Oct;26(10):1070-1; author reply 1071-2. Comment on: Nat Biotechnol. 2008 Jan;26(1):73-81. Allergenicity testing of GM crops. Aalberse RC. Publication Types: Comment Letter PMID: 18846066 [PubMed - indexed for MEDLINE] 59: Nat Biotechnol. 2008 Oct;26(10):1068-70. Comment on: Nat Biotechnol. 2008 Mar;26(3):247. Fuelling the 9 billion. Martindale W, Trewavas A. Publication Types: Comment Letter PMID: 18846065 [PubMed - indexed for MEDLINE] 60: Nat Biotechnol. 2008 Oct;26(10):1060. Plant biotech bonanza. Waltz E. Publication Types: News PMID: 18846061 [PubMed - indexed for MEDLINE] 61: Curr Biol. 2008 Sep 23;18(18):R839-41. Rift grows in GM debate. Williams N. PMID: 18843797 [PubMed - indexed for MEDLINE] 62: Environ Sci Pollut Res Int. 2008 Oct;15(7):529-35. Epub 2008 Oct 7. Hazard mitigation or mitigation hazard? Reuter H, Menzel G, Pehlke H, Breckling B. Department of General and Theoretical Ecology, Centre for Environmental Research and Sustainable Technology (UFT), University of Bremen, P. O. Box 330440, 28334 Bremen, Germany. hauke.reuter@zmt.uni-bremen.de BACKGROUND, AIM AND SCOPE: Transgenic oilseed rape (Brassica napus L.; OSR) is estimated to be environmentally and economically problematic because volunteers and ferals occur frequently and because of its hybridisation potential with several wild and weedy species. A proposed mitigation strategy aims to reduce survival, in particular in conventional OSR crops, by coupling the transgenic target modification with a dwarfing gene to reduce competitive fitness. Our study allowed us to access potential ecological implications of this strategy. MATERIALS AND METHODS: On a large scale (>500 km(2)), we recorded phenological and population parameters of oilseed rape plants for several years in rural and urban areas of Northern Germany (Bremen and surroundings). The characterising parameter were analysed for differences between wild and cultivated plants. RESULTS: In rural areas, occurrences of feral and volunteer OSR together had an average density of 1.19 populations per square kilometre, in contrast to urban areas where we found 1.68 feral populations per square kilometre on average. Throughout the survey, the vegetation cover at the locations with feral OSR ranged from less than 10% to 100%. Our investigations gave clear empirical evidence that feral OSR was, on average, at least 41% smaller than cultivated OSR, independent of phenological state after onset of flowering. DISCUSSION: The findings can be interpreted as phenotypic adaptation of feral OSR plants. Therefore, it must be asked whether dwarfing could be interpreted as an improvement of pre-adaptation to feral environments. In most of the sites where feral plants occurred, germination and establishment were in locations with disturbed vegetation cover, allowing initial growth without competition. Unless feral establishment of genetically modified dwarfed traits are specifically studied, it would not be safe to assume that the mitigation strategy of dwarfing also reduces dispersal in feral environments. CONCLUSIONS AND RECOMMENDATIONS: With respect to OSR, we argue that the proposed mitigation approach could increase escape and persistence of transgene varieties rather than reducing them. We conclude that the development of effective hazard mitigation measures in the risk evaluation of genetically modified organisms requires thorough theoretical and empirical ecological analyses rather than assumptions about abstract fitness categories that apply only in parts of the environment where the plant can occur. Publication Types: Research Support, Non-U.S. Gov't PMID: 18839232 [PubMed - indexed for MEDLINE] 63: Commun Agric Appl Biol Sci. 2008;73(1):27-31. Optimizing Arabidopsis thaliana as a platform for the seed-based production of recombinant proteins. Demeyer R, Vereecken F, De Loose M, Van Droogenbroeck B. Technology & Food Unit, Institute for Agricultural and Fisheries Research, Burg. Van Gansberghelaan 115, BE-9820, Merelbeke, Belgium. PMID: 18831240 [PubMed - indexed for MEDLINE] 64: Plant J. 2009 Feb;57(3):487-97. Epub 2008 Sep 26. TaASY1 promotes homologous chromosome interactions and is affected by deletion of Ph1. Boden SA, Langridge P, Spangenberg G, Able JA. Molecular Plant Breeding Cooperative Research Centre, School of Agriculture, Food and Wine, The University of Adelaide, Waite Campus, PMB1, Glen Osmond, South Australia, 5064, Australia. During meiosis, chromosomes are sorted into homologous pairs as a preface to their intimate association via recombination and synapsis. However, little is known about the mechanism used to distinguish homologous chromosomes from other chromosomes present in the nucleus. Studies in wheat (Triticum aestivum) have shown that the Pairing homoeologous 1 (Ph1) locus is required to suppress interactions between genetically similar homoeologous chromosomes. Here we show that absence of Ph1 causes increased transcription of Asynapsis 1 (ASY1), a gene that encodes an axial-element-associated protein that is essential for synapsis and cross-over formation in Arabidopsis and rice. Localisation of ASY1 during meiosis is also affected by deletion of Ph1. In addition, transgenic wheat mutants with decreased activity of TaASY1 display reduced synapsis during prophase I and exhibit pairing between homoeologous chromosomes at metaphase I. These results suggest that ASY1 is required to promote interactions between homologous chromosomes in bread wheat, and that Ph1 has a gene regulatory role, which is consistent with its suggested genetic identity as a Cdk-like gene. Broader implications of this research suggest that we could use the Taasy1 mutants to assess their efficacy in alien chromatin introgression studies, as seen with the ph1b mutant. Publication Types: Research Support, Non-U.S. Gov't PMID: 18826431 [PubMed - indexed for MEDLINE] 65: N Biotechnol. 2008;25(2-3):101-7. Epub 2008 Sep 6. Integrated farming: why organic farmers should use transgenic crops. Ammann K. Delft University of Technology, Julianalaan 67, NL-2628 BC Delft, Netherlands. klaus.ammann@ips.unibe.ch The concept of organic farming is summarised and compared as an example to farming with biotechnology-derived crops. If done within an ecological concept, both methods can be seen as environmentally acceptable. Organic farming does not offer consistent arguments for the rejection of transgenic crops. Some arguments (from genomics to biodiversity) are discussed in order to demonstrate that the contrast between both farming systems is rated too high and that it is possible to overcome the divide. In this way the ground is prepared for a proposal on how to merge those otherwise incompatible agricultural management systems, a proposal that also will have to build on a new concept of sustainability. It will be dealt with in the second part of the article in the next issue of New Biotechnology. PMID: 18824150 [PubMed - indexed for MEDLINE] 66: Proc Natl Acad Sci U S A. 2008 Sep 30;105(39):14826-31. Epub 2008 Sep 24. Control of feeding behavior in C. elegans by human G protein-coupled receptors permits screening for agonist-expressing bacteria. Teng MS, Shadbolt P, Fraser AG, Jansen G, McCafferty J. Wellcome Trust Sanger Institute, Hinxton, Cambridgeshire CB10 1HH, United Kingdom. G protein-coupled receptors (GPCRs) have a key role in many biological processes and are important drug targets for many human diseases. Therefore, understanding the molecular interactions between GPCRs and their ligands would improve drug design. Here, we describe an approach that allows the rapid identification of functional agonists expressed in bacteria. Transgenic Caenorhabditis elegans expressing the human chemokine receptor 5 (CCR5) in nociceptive neurons show avoidance behavior on encounter with the ligand MIP-1alpha and avoid feeding on Escherichia coli expressing MIP-1alpha compared with control bacteria. This system allows a simple activity screen, based on the distribution of transgenic worms in a binary food-choice assay, without a requirement for protein purification or tagging. By using this approach, a library of 68 MIP-1alpha variants was screened, and 13 critical agonist residues involved in CCR5 activation were identified, four of which (T8, A9, N22, and A25) have not been described previously, to our knowledge. Identified residues were subsequently validated in receptor binding assays and by calcium flux assays in mammalian cells. This approach serves not only for structure/function studies as demonstrated, but may be used to facilitate the discovery of agonists within bacterial libraries. Publication Types: Research Support, Non-U.S. Gov't PMID: 18815363 [PubMed - indexed for MEDLINE] 67: Plant J. 2009 Feb;57(3):413-25. Epub 2008 Sep 21. Refunctionalization of the ancient rice blast disease resistance gene Pit by the recruitment of a retrotransposon as a promoter. Hayashi K, Yoshida H. National Agricultural Research Center, National Agriculture and Food Research Organization, 1-2-1 Inada, Jo-etsu, Niigata 943 0193, Japan. The plant genome contains a large number of disease resistance (R) genes that have evolved through diverse mechanisms. Here, we report that a long terminal repeat (LTR) retrotransposon contributed to the evolution of the rice blast resistance gene Pit. Pit confers race-specific resistance against the fungal pathogen Magnaporthe grisea, and is a member of the nucleotide-binding site leucine-rich repeat (NBS-LRR) family of R genes. Compared with the non-functional allele Pit(Npb), the functional allele Pit(K59) contains four amino acid substitutions, and has the LTR retrotransposon Renovator inserted upstream. Pathogenesis assays using chimeric constructs carrying the various regions of Pit(K59) and Pit(Npb) suggest that amino acid substitutions might have a potential effect in Pit resistance; more importantly, the upregulated promoter activity conferred by the Renovator sequence is essential for Pit function. Our data suggest that transposon-mediated transcriptional activation may play an important role in the refunctionalization of additional 'sleeping' R genes in the plant genome. PMID: 18808453 [PubMed - indexed for MEDLINE] 68: Appl Spectrosc. 2008 Sep;62(9):1044-7. Discrimination of transgenic and conventional soybean seeds by fourier transform infrared photoacoustic spectroscopy. Caires AR, Teixeira MR, Súarez YR, Andrade LH, Lima SM. Grupo de Optica Aplicada, Universidade Federal da Grande Dourados-UFGD, C.P. 533, 79804-970, Dourados, MS, Brazil. andercaires@ufgd.edu.br Publication Types: Research Support, Non-U.S. Gov't PMID: 18801246 [PubMed - indexed for MEDLINE] 69: Prep Biochem Biotechnol. 2008;38(4):411-21. Expression of an HCV core antigen coding gene in tobacco (N. tabacum L.). Nianiou I, Kalantidis K, Madesis P, Georgopoulou U, Mavromara P, Tsaftaris A. Department of Genetics and Plant Breeding, School of Agriculture, Aristotelian University of Thessaloniki, Greece. Hepatitis C virus (HCV) is the major agent causing chronic liver disease. The core gene is the most conserved sequence in the HCV genome and proved immunoreactive when expressed in bacteria and antigenic in humans. In order to test the ability of plants to express the core gene for the production of core antigen, transgenic tobacco plants carrying the core gene were generated. The core protein was stably synthesized in T(0) and T(1) generations and was found to be immunoreactive, not only with anti-core polyclonal and monoclonal antibodies, but also was able to recognize the HCV virus in infected human serum. The prospects of producing a plant based vaccine and/or a food vaccine for this important virus are discussed. PMID: 18800303 [PubMed - indexed for MEDLINE] 70: Food Chem Toxicol. 2008 Aug 29. [Epub ahead of print] Timely awareness and prevention of emerging chemical and biochemical risks in foods: Proposal for a strategy based on experience with recent cases. Kleter GA, Groot MJ, Poelman M, Kok EJ, Marvin HJ. RIKILT - Institute of Food Safety, Wageningen University and Research Center, P.O. Box 230, NL-6700 AE Wageningen, The Netherlands. A number of recent food safety incidents have involved chemical substances, while various activities aim at the early identification of emerging chemical risks. This review considers recent cases of chemical and biochemical risks, as a basis for recommendations for awareness and prevention of similar risks at an early stage. These cases include examples of unapproved genetically modified food crops, intoxications with botanical products containing unintentionally admixed toxic herbs, residues of unapproved antibiotics and contaminants in farmed aquaculture species such as shrimp and salmon; and adverse effects of chemical and biological pesticides of natural origin. Besides case-specific recommendations for mitigation of future incidents of the same nature, general inferences and recommendations are made. It is recommended, for example, to establish databases for contaminants potentially present within products. Pro-active reconnaissance can facilitate the identification of products potentially contaminated with hazardous substances. In international trade, prevention and early identification of hazards are aided by management systems for product quality and safety, rigorous legislation, and inspections of consignments destined for export. Cooperation with the private sector and foreign authorities may be required to achieve these goals. While food and feed safety are viewed from the European perspective, the outcomes also apply to other regions. PMID: 18790713 [PubMed - as supplied by publisher] 71: Crit Rev Food Sci Nutr. 2008 Oct;48(9):799-823. Potato: a comparative study of the effect of cultivars and cultivation conditions and genetic modification on the physico-chemical properties of potato tubers in conjunction with multivariate analysis towards authenticity. Arvanitoyannis IS, Vaitsi O, Mavromatis A. School of Agricultural Sciences, Department of Agriculture Ichtyology and Aquatic Environment, University of Thessaly, Volos, Hellas, Greece. parmenion@uth.gr Potato (Solanum tuberosum L.) is a highly nutritious, mild flavored, easy to blend food that has many possibilities for "building in" desired nutrients. Varietal and environmental differences are known to exist in the shape, size, and nutritional content of potatoes. Different populations opt for varying sensory properties in relation to their diets. Potatoes are a low energy food in comparison to cereals and legumes. The aim of this review was to present an update of the currently conducted studies both on the characterization of several potato varieties (physical, chemical, and sensory analysis) and by means of genetic modification. Towards this target, five comprehensive tables were compiled where all recent data (physicochemical properties) and GM varieties were presented in conjunction with multivariate analysis (chemometrics). The latter was shown to be effectively used towards authenticity purposes (identification of geographical origin, variety, GM). Publication Types: Review PMID: 18788007 [PubMed - indexed for MEDLINE] 72: Shokuhin Eiseigaku Zasshi. 2008 Aug;49(4):272-82. A 104-week feeding study of genetically modified soybeans in F344 rats. Sakamoto Y, Tada Y, Fukumori N, Tayama K, Ando H, Takahashi H, Kubo Y, Nagasawa A, Yano N, Yuzawa K, Ogata A. Department of Environmental Health and Toxicology, Tokyo Metropolitan Institute of Public Health. Tokyo, Japan. A chronic feeding study to evaluate the safety of genetically modified glyphosate-tolerant soybeans (GM soybeans) was conducted using F344 DuCrj rats. The rats were fed diet containing GM soybeans or Non-GM soybeans at the concentration of 30% in basal diet. Non-GM soybeans were a closely related strain to the GM soybeans. These two diets were adjusted to an identical nutrient level. In this study, the influence of GM soybeans in rats was compared with that of the Non-GM soybeans, and furthermore, to assess the effect of soybeans themselves, the groups of rats fed GM and Non-GM soybeans were compared with a group fed commercial diet (CE-2). General conditions were observed daily and body weight and food consumption were recorded. At the termination (104 weeks), animals were subjected to hematology, serum biochemistry, and pathological examinations. There were several differences in animal growth, food intake, organ weights and histological findings between the rats fed the GM and/or Non-GM soybeans and the rats fed CE-2. However, body weight and food intake were similar for the rats fed the GM and Non-GM soybeans. Gross necropsy findings, hematological and serum biochemical parameters, and organ weights showed no meaningful difference between rats fed the GM and Non-GM soybeans. In pathological observation, there was neither an increase in incidence nor any specific type of nonneoplastic or neoplastic lesions in the GM soybeans group in each sex. These results indicate that long-term intake of GM soybeans at the level of 30% in diet has no apparent adverse effect in rats. PMID: 18787312 [PubMed - in process] 73: Natl Toxicol Program Genet Modif Model Rep. 2007 Dec;(12):1-85. The toxicology and carcinogenesis study of phenolphthalein (CAS No. 77-09-8) in genetically modified haploinsufficient p16(Ink4a)/p19(Arf) mice (feed study). [No authors listed] Phenolphthalein was commonly used as a laxative for most of the 20th century. The use of phenolphthalein in laxatives has decreased since 1997 when the United States Food and Drug Administration (FDA) proposed to withdraw its classification as an over-the-counter drug (21 CFR, Part 310). Phenolphthalein has been previously evaluated in 2-year carcinogenicity studies by the National Toxicology Program (1996). The major route of human exposure to phenolphthalein is via ingestion, dermal contact, and inhalation of contaminated air originating from process units manufacturing the compound. In this study, the carcinogenic effects of phenolphthalein were studied in the haploinsufficient p16(Ink4a)/p19(Arf) mouse model as an ongoing goal of the NTP is to seek model systems for toxicology and carcinogenesis studies, especially those that can provide mechanistic information relative to understanding an agent's mode of action. Male and female haploinsufficient p16(Ink4a)/p19(Arf) mice were exposed to phenolphthalein (greater than 97% pure) in feed for 27 weeks. Genetic toxicology studies were conducted in mouse peripheral blood erythrocytes. 27-WEEK STUDY IN MICE: Groups of 15 male and 15 female mice were exposed to 0, 200, 375, 750, 3,000, or 12,000 ppm phenolphthalein (equivalent to average daily doses of approximately 35, 65, 135, 540, and 2,170 mg phenolphthalein/kg body weight to males and 50, 90, 170, 680, 2,770 mg/kg to females) in feed for 27 weeks. Survival of all exposed groups of male and female mice was similar to that of the control groups. Mean body weights of males in the 12,000 ppm group were less than those of the control group after week 11. No differences in feed consumption were noted between exposed and control groups. Atypical hyperplasia of the thymus, a premalignant change of chemically induced thymic lymphoma, occurred in exposed males and females, and the incidence was significantly increased in 12,000 ppm females. Atrophy of the seminiferous tubules in the testis, hyperplasia of the testicular interstitial (Leydig) cells, and epididymal hypospermia occurred in most 3,000 and 12,000 ppm males. Additionally, the left and right testis weights, the left epididymis weights, sperm motility, the numbers of spermatid heads per testis, and sperm heads per cauda and per gram cauda epididymis were generally significantly less in 3,000 and 12,000 ppm males than in the control group. The incidences of nephropathy were significantly increased in 3,000 and 12,000 ppm males; incidences of hypertrophy of renal tubules were significantly increased in males receiving 750 ppm or greater. Hematopoietic cell proliferation of the spleen occurred in all 12,000 ppm males, and the incidences of this lesion were significantly increased in 375, 750, and 12,000 ppm females. GENETIC TOXICOLOGY: The frequency of micronucleated erythrocytes was assessed at four time points during the 27-week study in male and female haploinsufficient p16(Ink4a)/p19(Arf) mice. Significant concentration-related increases in micronucleated cells were observed at all time points in male and female mice. CONCLUSIONS: Under the conditions of this 27-week feed study, there was no evidence of carcinogenic activity of phenolphthalein in male or female haploinsufficient p16(Ink4a)/p19(Arf) mice exposed to 200, 375, 750, 3,000, or 12,000 ppm. Because this is a new model, there is uncertainty whether the study possessed sufficient sensitivity to detect a carcinogenic effect. Phenolphthalein induced atypical hyperplasia, a preneoplastic lesion of the thymus, in male and female mice, hematopoietic cell proliferation of the spleen in male and female mice, and toxicity to the kidney and reproductive system in male mice. PMID: 18784766 [PubMed - indexed for MEDLINE] 74: Natl Toxicol Program Genet Modif Model Rep. 2005 Oct;(2):1-113. NTP toxicology studies of acesulfame potassium (CAS No. 55589-62-3) in genetically modified (FVB Tg.AC Hemizygous) mice and carcinogenicity studies of acesulfame potassium in genetically modified [B6.129-Trp53(tm1Brd) (N5) Haploinsufficient] mice (feed studies)mice. [No authors listed] Acesulfame potassium is an artificial sweetener used throughout the world in food and beverages. Acesulfame potassium was nominated by The Center for Science in the Public Interest because of its widespread use. Male and female Tg.AC hemizygous and p53 haploinsufficient mice were exposed to acesulfame potassium (at least 99% pure) in feed for 9 months. Genetic toxicology studies were conducted in mouse peripheral blood erythrocytes. 9-MONTH STUDY IN Tg.AC HEMIZYGOUS MICE: Groups of 15 male and 15 female Tg.AC hemizygous mice were fed diets containing 0%, 0.3%, 1%, or 3% acesulfame potassium (equivalent to average daily doses of approximately 420, 1,400, or 4,500 mg acesulfame potassium/kg body weight to males and 520, 1,700, or 5,400 mg/kg to females) for 40 weeks. Exposure to acesulfame potassium had no effect on survival or mean body weights. Feed consumption by the exposed groups was similar to that by the control groups throughout the study. There were no neoplasms or nonneoplastic lesions that were attributed to exposure to acesulfame potassium. 9-MONTH STUDY IN p53 HAPLOINSUFFICIENT MICE: Groups of 15 male and 15 female p53 haploinsufficient mice were fed diets containing 0%, 0.3%, 1%, or 3% acesulfame potassium (equivalent to average daily doses of approximately 475, 1,500, or 4,700 mg/kg to males and 570, 1,800, or 5,700 mg/kg to females) for 40 weeks. Exposure to acesulfame potassium had no effect on survival or mean body weights. Feed consumption by the exposed groups was similar to that by the control groups throughout the study. There were no neoplasms or nonneoplastic lesions that were attributed to exposure to acesulfame potassium. GENETIC TOXICOLOGY: Acesulfame potassium did not increase the frequency of micronucleated erythrocytes in peripheral blood of male or female Tg.AC hemizygous mice administered 0.3% to 3% in dosed feed. A similar study was conducted in p53 haploinsufficient mice, and a significant exposure concentration-related increase in the frequency of micronucleated erythrocytes was noted in males but not females. CONCLUSIONS: Under the conditions of this 9-month feed study, there was no evidence of carcinogenic activity of acesulfame potassium in male or female p53 haploinsufficient mice exposed to 0.3%, 1%, or 3%. PMID: 18784762 [PubMed - indexed for MEDLINE] 75: BMC Plant Biol. 2008 Sep 10;8:91. Transcriptional profiling of pea ABR17 mediated changes in gene expression in Arabidopsis thaliana. Krishnaswamy SS, Srivastava S, Mohammadi M, Rahman MH, Deyholos MK, Kav NN. Department of Agricultural, Food and Nutritional Science, University of Alberta, Edmonton, AB T6G 2P5, Canada. krishnas@ualberta.ca BACKGROUND: Pathogenesis-related proteins belonging to group 10 (PR10) are elevated in response to biotic and abiotic stresses in plants. Previously, we have shown a drastic salinity-induced increase in the levels of ABR17, a member of the PR10 family, in pea. Furthermore, we have also demonstrated that the constitutive expression of pea ABR17 cDNA in Arabidopsis thaliana and Brassica napus enhances their germination and early seedling growth under stress. Although it has been reported that several members of the PR10 family including ABR17 possess RNase activity, the exact mechanism by which the aforementioned characteristics are conferred by ABR17 is unknown at this time. We hypothesized that a study of differences in transcriptome between wild type (WT) and ABR17 transgenic A. thaliana may shed light on this process. RESULTS: The molecular changes brought about by the expression of pea ABR17 cDNA in A. thaliana in the presence or absence of salt stress were investigated using microarrays consisting of 70-mer oligonucleotide probes representing 23,686 Arabidopsis genes. Statistical analysis identified number of genes which were over represented among up- or down-regulated transcripts in the transgenic line. Our results highlight the important roles of many abscisic acid (ABA) and cytokinin (CK) responsive genes in ABR17 transgenic lines. Although the transcriptional changes followed a general salt response theme in both WT and transgenic seedlings under salt stress, many genes exhibited differential expression patterns when the transgenic and WT lines were compared. These genes include plant defensins, heat shock proteins, other defense related genes, and several transcriptional factors. Our microarray results for selected genes were validated using quantitative real-time PCR. CONCLUSION: Transcriptional analysis in ABR17 transgenic Arabidopsis plants, both under normal and saline conditions, revealed significant changes in abundance of transcripts for many stress responsive genes, as well as those related to plant growth and development. Our results also suggest that ABR17 may mediate stress tolerance through the modulation of many ABA- and CK-responsive genes and may further our understanding of the role of ABR17 in mediating plant stress responses. Publication Types: Research Support, Non-U.S. Gov't PMID: 18783601 [PubMed - indexed for MEDLINE] 76: J Agric Food Chem. 2008 Oct 8;56(19):9206-14. Epub 2008 Sep 10. Assessment of the nutritional values of genetically modified wheat, corn, and tomato crops. Venneria E, Fanasca S, Monastra G, Finotti E, Ambra R, Azzini E, Durazzo A, Foddai MS, Maiani G. Istituto Nazionale di Ricerca per gli Alimenti e la Nutrizione, 00178 Roma, Italy. venneria@inran.it The genetic modification in fruit and vegetables could lead to changes in metabolic pathways and, therefore, to the variation of the molecular pattern, with particular attention to antioxidant compounds not well-described in the literature. The aim of the present study was to compare the quality composition of transgenic wheat ( Triticum durum L.), corn ( Zea mays L.), and tomato ( Lycopersicum esculentum Mill.) to the nontransgenic control with a similar genetic background. In the first experiment, Ofanto wheat cultivar containing the tobacco rab1 gene and nontransgenic Ofanto were used. The second experiment compared two transgenic lines of corn containing Bacillus thuringiensis "Cry toxin" gene (PR33P67 and Pegaso Bt) to their nontransgenic forms. The third experiment was conducted on transgenic tomato ( Lycopersicum esculentum Mill.) containing the Agrobacterium rhizogenes rolD gene and its nontransgenic control (cv. Tondino). Conventional and genetically modified crops were compared in terms of fatty acids content, unsaponifiable fraction of antioxidants, total phenols, polyphenols, carotenoids, vitamin C, total antioxidant activity, and mineral composition. No significant differences were observed for qualitative traits analyzed in wheat and corn samples. In tomato samples, the total antioxidant activity (TAA), measured by FRAP assay, and the naringenin content showed a lower value in genetically modified organism (GMO) samples (0.35 mmol of Fe (2+) 100 g (-1) and 2.82 mg 100 g (-1), respectively), in comparison to its nontransgenic control (0.41 mmol of Fe (2+) 100 g (-1) and 4.17 mg 100 g (-1), respectively). On the basis of the principle of substantial equivalence, as articulated by the World Health Organization, the Organization for Economic Cooperation and Development, and the United Nations Food and Agriculture Organization, these data support the conclusion that GM events are nutritionally similar to conventional varieties of wheat, corn, and tomato on the market today. PMID: 18781763 [PubMed - indexed for MEDLINE] 77: Nat Biotechnol. 2008 Sep;26(9):975-8. Trace and traceability--a call for regulatory harmony. Ramessar K, Capell T, Twyman RM, Quemada H, Christou P. Publication Types: Letter PMID: 18779799 [PubMed - indexed for MEDLINE] 78: Nat Biotechnol. 2008 Sep;26(9):974-5. Comment in: Nat Biotechnol. 2008 Dec;26(12):1335; author reply 1335. Auf Wiedersehen, agbiotech. Miller HI. Publication Types: Letter PMID: 18779798 [PubMed - indexed for MEDLINE] 79: J R Soc Med. 2008 Sep;101(9):435. Comment on: J R Soc Med. 2008 Jun;101(6):290-8. Errors in text. Schubert D. Publication Types: Comment Letter PMID: 18779242 [PubMed - indexed for MEDLINE] 80: Regul Toxicol Pharmacol. 2008 Dec;52(3):208-22. Epub 2008 Aug 22. A risk-based classification scheme for genetically modified foods. I: Conceptual development. Chao E, Krewski D. McLaughlin Centre for Population Health Risk Assessment, Institute of Population Health, University of Ottawa, 1 Stewart Street, Ottawa, Ont., Canada KIN 6N5. echao@uottawa.ca The predominant paradigm for the premarket assessment of genetically modified (GM) foods reflects heightened public concern by focusing on foods modified by recombinant deoxyribonucleic acid (rDNA) techniques, while foods modified by other methods of genetic modification are generally not assessed for safety. To determine whether a GM product requires less or more regulatory oversight and testing, we developed and evaluated a risk-based classification scheme (RBCS) for crop-derived GM foods. The results of this research are presented in three papers. This paper describes the conceptual development of the proposed RBCS that focuses on two categories of adverse health effects: (1) toxic and antinutritional effects, and (2) allergenic effects. The factors that may affect the level of potential health risks of GM foods are identified. For each factor identified, criteria for differentiating health risk potential are developed. The extent to which a GM food satisfies applicable criteria for each factor is rated separately. A concern level for each category of health effects is then determined by aggregating the ratings for the factors using predetermined aggregation rules. An overview of the proposed scheme is presented, as well as the application of the scheme to a hypothetical GM food. Publication Types: Research Support, Non-U.S. Gov't PMID: 18778747 [PubMed - indexed for MEDLINE] 81: Regul Toxicol Pharmacol. 2008 Dec;52(3):223-34. Epub 2008 Aug 15. A risk-based classification scheme for genetically modified foods. II: Graded testing. Chao E, Krewski D. McLaughlin Centre for Population Health Risk Assessment, Institute of Population Health, University of Ottawa, 1 Stewart Street, Ottawa, Ont., Canada K1N 6N5. echao@uottawa.ca This paper presents a graded approach to the testing of crop-derived genetically modified (GM) foods based on concern levels in a proposed risk-based classification scheme (RBCS) and currently available testing methods. A graded approach offers the potential for more efficient use of testing resources by focusing less on lower concern GM foods, and more on higher concern foods. In this proposed approach to graded testing, products that are classified as Level I would have met baseline testing requirements that are comparable to what is widely applied to premarket assessment of GM foods at present. In most cases, Level I products would require no further testing, or very limited confirmatory analyses. For products classified as Level II or higher, additional testing would be required, depending on the type of the substance, prior dietary history, estimated exposure level, prior knowledge of toxicity of the substance, and the nature of the concern related to unintended changes in the modified food. Level III testing applies only to the assessment of toxic and antinutritional effects from intended changes and is tailored to the nature of the substance in question. Since appropriate test methods are not currently available for all effects of concern, future research to strengthen the testing of GM foods is discussed. Publication Types: Research Support, Non-U.S. Gov't PMID: 18768151 [PubMed - indexed for MEDLINE] 82: J Econ Entomol. 2008 Aug;101(4):1134-9. Effects of Bt transgenic Chinese cabbage on the herbivore Mamestra brassicae (Lepidoptera: Noctuidae) and its parasitoid Microplitis mediator (Hymenoptera: Braconidae). Kim YH, Kang JS, Kim JI, Kwon M, Lee S, Cho HS, Lee SH. Department of Applied Biology and Chemistry, Seoul National University, Gwanak-gu, Seoul, South Korea. We investigated the effects of a diamondback moth-resistant Chinese cabbage (Brassica campestris subsp napus variety pekinensis Makino), expressing the insecticidal protein CrylA(c) toxin derived from Bacillus thuringiensis, on the nontarget herbivore Mamestra brassicae (L.) (Lepidoptera: Noctuidae) and its parasitoid wasp Microplitis mediator (Haliday) (Hymenoptera: Braconidae). A decreased survival rate at neonate stage was observed in M. brassicae when reared on Bt cabbage, although overall development was not significantly affected. According to enzyme-linked immunosorbent assay test using CrylA(c) antibody, the Cry toxin was only detected in the alimentary canal, not in the hemolymph or remaining body parts of M. brassicae, indicating that the ingested Cry toxin is neither distributed inside the body nor transferred through the trophic level. As expected, no Cry toxin was found in the larvae and cocoons of M. mediator. In addition, no significant changes were observed in the parasitization rate, larval period, pupal period, cocoon weight, or adult emergence rate when M. mediator wasps were reared on the M. brassicae larvae fed with transgenic Chinese cabbage. In summary, no direct or indirect adverse effects of transgenic Chinese cabbage on the two nontarget insect species were observed, suggestive of low risk in herbivore-parasitoid food chain. Publication Types: Research Support, Non-U.S. Gov't PMID: 18767720 [PubMed - indexed for MEDLINE] 83: Regul Toxicol Pharmacol. 2008 Dec;52(3):235-41. Epub 2008 Aug 15. A risk-based classification scheme for genetically modified foods. III: Evaluation using a panel of reference foods. Chao E, Krewski D. McLaughlin Centre for Population Health Risk Assessment, Institute of Population Health, University of Ottawa, 1 Stewart Street, Ottawa, Ont., Canada K1N 6N5. echao@uottawa.ca This paper presents an exploratory evaluation of four functional components of a proposed risk-based classification scheme (RBCS) for crop-derived genetically modified (GM) foods in a concordance study. Two independent raters assigned concern levels to 20 reference GM foods using a rating form based on the proposed RBCS. The four components of evaluation were: (1) degree of concordance, (2) distribution across concern levels, (3) discriminating ability of the scheme, and (4) ease of use. At least one of the 20 reference foods was assigned to each of the possible concern levels, demonstrating the ability of the scheme to identify GM foods of different concern with respect to potential health risk. There was reasonably good concordance between the two raters for the three separate parts of the RBCS. The raters agreed that the criteria in the scheme were sufficiently clear in discriminating reference foods into different concern levels, and that with some experience, the scheme was reasonably easy to use. Specific issues and suggestions for improvements identified in the concordance study are discussed. Publication Types: Research Support, Non-U.S. Gov't PMID: 18765265 [PubMed - indexed for MEDLINE] 84: EMBO Rep. 2008 Sep;9(9):832-4. Up to the challenge? Rising prices for food and oil could herald a renaissance of plant science. Breithaupt H. PMID: 18762773 [PubMed - indexed for MEDLINE] 85: Plant Cell Rep. 2008 Nov;27(11):1741-54. Epub 2008 Aug 30. Efficient production of genetically engineered, male-sterile Arabidopsis thaliana using anther-specific promoters and genes derived from Brassica oleracea and B. rapa. Konagaya K, Ando S, Kamachi S, Tsuda M, Tabei Y. Division of Plant Science, National Institute of Agrobiological Sciences, Ibaraki, Japan. Prevention of transgene flow from genetically modified crops to food crops and wild relatives is of concern in agricultural biotechnology. We used genes derived from food crops to produce complete male sterility as a strategy for gene confinement as well as to reduce the food purity concerns of consumers. Anther-specific promoters (A3, A6, A9, MS2, and MS5) were isolated from Brassica oleracea and B. rapa and fused to the beta-glucuronidase (GUS) reporter gene and candidate genes for male sterility, including the cysteine proteases BoCysP1 and BoCP3, and negative regulatory components of phytohormonal responses involved in male development. These constructs were then introduced into Arabidopsis thaliana. GUS analyses revealed that A3, A6, and A9 had tapetum-specific promoter activity from the anther meiocyte stage. Male sterility was confirmed in tested constructs with protease or gibberellin insensitive (gai) genes. In particular, constructs with BoCysP1 driven by the A3 or A9 promoter most efficiently produced plants with complete male sterility. The tapetum and middle layer cells of anthers expressing BoCysP1 were swollen and excessively vacuolated when observed in transverse section. This suggests that the ectopic expression of cysteine protease in the meiocyte stage may inhibit programmed cell death. The gai gene also induced male sterility, although at a low frequency. This is the first report to show that plant cysteine proteases and gai from food crops are available as a novel tool for the development of genetically engineered male-sterile plants. Publication Types: Research Support, Non-U.S. Gov't PMID: 18758783 [PubMed - indexed for MEDLINE] 86: Food Chem Toxicol. 2008 Oct;46 Suppl 10:S15-9. Epub 2008 Aug 8. Analytical criteria for performance characteristics of IgE binding methods for evaluating safety of biotech food products. Holzhauser T, Ree R, Poulsen LK, Bannon GA. Division of Allergology, Paul-Ehrlich-Institut, Paul-Ehrlich-Strasse 51-59, 63225 Langen, Germany. holth@pei.de There is detailed guidance on how to perform bioinformatic analyses and enzymatic degradation studies for genetically modified crops under consideration for approval by regulatory agencies; however, there is no consensus in the scientific community on the details of how to perform IgE serum studies. IgE serum studies are an important safety component to acceptance of genetically modified crops when the introduced protein is novel, the introduced protein is similar to known allergens, or the crop is allergenic. In this manuscript, we describe the characteristics of the reagents, validation of assay performance, and data analysis necessary to optimize the information obtained from serum testing of novel proteins and genetically modified (GM) crops and to make results more accurate and comparable between different investigations. PMID: 18727951 [PubMed - indexed for MEDLINE] 87: J AOAC Int. 2008 Jul-Aug;91(4):957-64. Biopharming to increase bioactive peptides in rice seed. Yang L, Wakasa Y, Takaiwa F. Transgenic Crop Research and Development Center, National Institute of Agrobiological Sciences, Kannondai 2-1-2, Tsukuba Ibaraki 305-8602, Japan. The production of high-value pharmaceutical proteins and peptides in transgenic plants is an attractive and economically feasible alternative to conventional mammalian cell, yeast, and bacterial systems. In contrast to vegetative tissues, rice seeds allow higher accumulation of recombinant proteins and long-term stable storage. Rice is not only consumed as a staple food by a majority of the world's population, but is also used as a model monocot for biotechnological manipulation. Daily oral consumption of transgenic rice seeds that accumulate high concentrations of food-derived or synthetic bioactive peptides can be expected to provide a safe, reliable, and consistent oral delivery system that would contribute to the promotion of human health care. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 18727558 [PubMed - indexed for MEDLINE] 88: Anal Bioanal Chem. 2008 Oct;392(3):369-84. Epub 2008 Aug 23. Methods for detection of GMOs in food and feed. Marmiroli N, Maestri E, Gullì M, Malcevschi A, Peano C, Bordoni R, De Bellis G. Department of Environmental Sciences, Division of Genetics and Environmental Biotechnology, University of Parma, Viale G.P. Usberti 11A, 43100, Parma, Italy. This paper reviews aspects relevant to detection and quantification of genetically modified (GM) material within the feed/food chain. The GM crop regulatory framework at the international level is evaluated with reference to traceability and labelling. Current analytical methods for the detection, identification, and quantification of transgenic DNA in food and feed are reviewed. These methods include quantitative real-time PCR, multiplex PCR, and multiplex real-time PCR. Particular attention is paid to methods able to identify multiple GM events in a single reaction and to the development of microdevices and microsensors, though they have not been fully validated for application. Publication Types: Review PMID: 18726090 [PubMed - indexed for MEDLINE] 89: J Food Prot. 2008 Aug;71(8):1673-8. Comparison of Shiga-like toxin II expression between two genetically diverse lineages of Escherichia coli O157:H7. Dowd SE, Williams JB. U.S. Department of Agriculture, Agricultural Research Service Livestock Issues Research Unit, 1604 West FM 1294, Lubbock, Texas 79403, USA. sdowd@lbk.ars.usda.gov The existence of two separate lineages of Escherichia coli O157:H7 has previously been reported, and research indicates that one of these lineages (lineage I) might be more pathogenic toward human hosts. We postulated that the lineage more pathogenic expresses higher levels of Shiga toxin 2 (Stx2) than do the nonpathogenic lineage II. A comprehensive set of methodologies were used to investigate the difference in Stx2 protein and mRNA expression between the two lineages. An initial Stx2-specific enzyme-linked immunosorbent assay was conducted, and lineage I overall demonstrated significantly more toxin proteins expressed (P < 0.01). Gene expression analyses all showed significantly higher stx2 gene expression in lineage I (P = 0.02). PCR mapping revealed a possible explanation for decreased amounts of stx2 transcripts in the potentially nonpathogenic lineage II isolates, suggesting that genomic changes have modified the toxin-encoding region of the phage. This study provides additional data to support the existence of two diverse lineages of E. coli O157:H7, one of which may have lower pathogenic potential in relation to human hosts. The PCR described also provides a possible screening tool for E. coli O157 populations to differentiate these lineages. This study provides useful information on the ecology of E. coli O157, with broad implications within the clinical, scientific, and livestock industries. PMID: 18724763 [PubMed - indexed for MEDLINE] 90: J Med Food. 2008 Dec;11(4):601-5. The problem with nutritionally enhanced plants. Schubert DR. Cellular Neurobiology Laboratory, The Salk Institute for Biological Studies, La Jolla, California 92037-1099, USA. schubert@salk.edu Among the next generation of genetically modified (GM) plants are those that are engineered to produce elevated levels of nutritional molecules such as vitamins, omega-3 fatty acids, and amino acids. Based upon the U.S. current regulatory scheme, the plants and their products may enter our food supply without any required safety testing. The potential risks of this type of GM plant are discussed in the context of human health, and it is argued that there should be very careful safety testing of plants designed to produce biologically active molecules before they are commercially grown and consumed. This will require a mandatory, scientifically rigorous review process. PMID: 18721071 [PubMed - indexed for MEDLINE] 91: Obesity (Silver Spring). 2008 Oct;16(10):2289-95. Epub 2008 Jul 24. Variation in the bitter-taste receptor gene TAS2R38, and adiposity in a genetically isolated population in Southern Italy. Tepper BJ, Koelliker Y, Zhao L, Ullrich NV, Lanzara C, d'Adamo P, Ferrara A, Ulivi S, Esposito L, Gasparini P. Department of Food Science, School of Environmental and Biological Sciences, Rutgers University, New Brunswick, New Jersey, USA. tepper@aesop.rutgers.edu OBJECTIVE: Variation in the bitter-taste receptor gene, TAS2R38 confers the ability to taste 6-n-propylthiouracil (PROP). The objective of this study was to relate TAS2R38 haplotypes and PROP-tasting phenotypes to adiposity in a genetically isolated population. We hypothesized that the nontaster phenotype would be associated with higher BMI and waist circumference (WC) in females, and that dietary restraint would mediate this relationship. METHODS AND PROCEDURES: Participants were 540 healthy inhabitants of the genetically isolated village of Carlantino in southern Italy who were 15-89 years of age at the time of the study. Haplotype analyses were performed and PROP tasting was assessed using a filter paper method. Height, weight, and WC were measured and restrained eating was assessed using a brief questionnaire. RESULTS: Nontaster females had higher BMI and WC than females who were phenotypic tasters, and this relationship was specific to females with low dietary restraint. Regression analysis showed that BMI declined by 1.7 units across taster groups in females when the model included the PROP by restraint interaction. PROP phenotype was not significantly associated with WC in the regression models. Polymorphisms in TAS2R38 were not associated with BMI or WC in females. Neither TAS2R38 haplotype nor PROP phenotype was strongly related to BMI or WC in males. DISCUSSION: These data support previous findings of a relation between the nontaster phenotype and higher BMI in females that is modified by dietary restraint. Assessment of PROP phenotypes might provide unique information about adiposity that is not captured by haplotype analysis alone. Publication Types: Research Support, Non-U.S. Gov't PMID: 18719631 [PubMed - indexed for MEDLINE] 92: Food Chem Toxicol. 2008 Oct;46 Suppl 10:S24-34. Epub 2008 Jul 31. Performing IgE serum testing due to bioinformatics matches in the allergenicity assessment of GM crops. Goodman RE. Food Allergy Research and Resource Program, Department of Food Science and Technology, University of Nebraska, 143 Food Industry Complex, Lincoln, USA. rgoodman2@unl.edu Proteins introduced into genetically modified (GM) organisms through genetic engineering must be evaluated for their potential to cause allergic disease under various national laws and regulations. The Codex Alimentarius Commission guidance document (2003) calls for testing of serum IgE binding to the introduced protein if the gene was from an allergenic source, or the sequence of the transferred protein has >35% identity in any segment of 80 or more amino acids to a known allergen or shares significant short amino acid identities. The Codex guidance recognized that the assessment will evolve based on new scientific knowledge. Arguably, the current criteria are too conservative as discussed in this paper and they do not provide practical guidance on serum testing. The goals of this paper are: (1) to summarize evidence supporting the level of identity that indicates potential risk of cross-reactivity for those with existing allergies; (2) to provide example bioinformatics results and discuss their interpretation using published examples of proteins expressed in transgenic crops; and (3) to discuss key factors of experimental design and methodology for serum IgE tests to minimize the rate of false negative and false positive identification of potential allergens and cross-reactive proteins. PMID: 18715545 [PubMed - indexed for MEDLINE] 93: J Integr Plant Biol. 2008 Aug;50(8):991-6. Lycopene accumulation affects the biosynthesis of some carotenoid-related volatiles independent of ethylene in tomato. Gao H, Zhu H, Shao Y, Chen A, Lu C, Zhu B, Luo Y. College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, China. For elucidating the regulatory mechanism of ethylene on carotenoid-related volatiles (open chain) compounds and the relationship between lycopene and carotenoid-related volatiles, transgenic tomato fruits in which ACC synthase was suppressed were used. The transgenic tomato fruit showed a significant reduction of lycopene and aroma volatiles with low ethylene production. 6-Methyl-5-hepten-2-one, 6-methyl-5-hepten-2-ol and geranylacetone, which were suspected to be lycopene degradation products, were lower than those in wild type tomato fruits. In order to identify whether lycopene accumulation effects the biosynthesis of some carotenoid-related volatiles independent of ethylene in tomato or not, the capability of both wild type and transgenic tomato fruits discs to convert lycopene into carotenoid-related volatiles was evaluated. The data showed that external lycopene could convert into 6-methyl-5-hepten-2-one and 6-methyl-5-hepten-2-ol in vivo, indicating that the strong inhibition of ethylene production had no effect on enzymes in the biosynthesis pathway of some carotenoid-related volatiles. Therefore, in ACS-suppression transgenic tomato fruits, the low levels of 6-methyl-5-hepten-2-one, 6-methyl-5-hepten-2-ol was due to decreased lycopene accumulation, not ethylene production. Ethylene only affected the accumulation of lycopene, and then indirectly influenced the level of lycopene-related volatiles. Publication Types: Research Support, Non-U.S. Gov't PMID: 18713349 [PubMed - indexed for MEDLINE] 94: Curr Biol. 2008 Jul 22;18(14):R575-6. Spanish farmers welcome GM maize. Williams N. Publication Types: News PMID: 18711794 [PubMed - indexed for MEDLINE] 95: Nucleic Acids Res. 2008 Oct;36(18):e118. Epub 2008 Aug 18. NAIMA: target amplification strategy allowing quantitative on-chip detection of GMOs. Morisset D, Dobnik D, Hamels S, Zel J, Gruden K. Department of Biotechnology and Systems Biology, National Institute of Biology, Vecna pot 111, Ljubljana 1000, Slovenia and Eppendorf Array Technologies SA, Rue du séminaire 20, B-5000 Namur, Belgium. dany.morisset@nib.si We have developed a novel multiplex quantitative DNA-based target amplification method suitable for sensitive, specific and quantitative detection on microarray. This new method named NASBA Implemented Microarray Analysis (NAIMA) was applied to GMO detection in food and feed, but its application can be extended to all fields of biology requiring simultaneous detection of low copy number DNA targets. In a first step, the use of tailed primers allows the multiplex synthesis of template DNAs in a primer extension reaction. A second step of the procedure consists of transcription-based amplification using universal primers. The cRNA product is further on directly ligated to fluorescent dyes labelled 3DNA dendrimers allowing signal amplification and hybridized without further purification on an oligonucleotide probe-based microarray for multiplex detection. Two triplex systems have been applied to test maize samples containing several transgenic lines, and NAIMA has shown to be sensitive down to two target copies and to provide quantitative data on the transgenic contents in a range of 0.1-25%. Performances of NAIMA are comparable to singleplex quantitative real-time PCR. In addition, NAIMA amplification is faster since 20 min are sufficient to achieve full amplification. Publication Types: Evaluation Studies Research Support, Non-U.S. Gov't PMID: 18710880 [PubMed - indexed for MEDLINE] 96: Anal Bioanal Chem. 2008 Oct;392(3):327-31. The genetically modified foods debate: demystifying the controversy through analytical chemistry. Daunert S, Deo S, Morin X, Roda A. Publication Types: Editorial Introductory Journal Article PMID: 18709361 [PubMed - indexed for MEDLINE] 97: Food Chem Toxicol. 2008 Oct;46 Suppl 10:S20-3. Epub 2008 Jul 30. Current codex guidelines for assessment of potential protein allergenicity. Ladics GS. DuPont Experimental Station, Building 353, Wilmington, DE 19880, USA. Gregory.s.ladics@usa.dupont.com A rigorous safety assessment process exists for GM crops. It includes evaluation of the introduced protein as well as the crop containing such protein with the goal of demonstrating the GM crop is "as-safe-as" non-transgenic crops in the food supply. One of the major issues for GM crops is the assessment of the expressed protein for allergenic potential. Currently, no single factor is recognized as an identifier for protein allergenicity. Therefore, a weight-of-evidence approach, which takes into account a variety of factors and approaches for an overall assessment of allergenic potential, is conducted [Codex Alimentarious Commission, 2003. Alinorm 03/34: Joint FAO/WHO Food Standard Programme, Codex Alimentarious Commission, Twenty-Fifth Session, Rome, Italy, 30 June-5 July, 2003. Appendix III, Guideline for the conduct of food safety assessment of foods derived from recombinant-DNA plants, and Appendix IV, Annex on the assessment of possible allergenicity, pp. 47-60]. This assessment is based on what is known about allergens, including the history of exposure and safety of the gene(s) source; protein structure (e.g., amino acid sequence identity to human allergens); stability to pepsin digestion in vitro [Thomas, K. et al., 2004. A multi-laboratory evaluation of a common in vitro pepsin digestion assay protocol used in assessing the safety of novel proteins. Regul. Toxicol. Pharmacol. 39, 87-98]; an estimate of exposure of the novel protein(s) to the gastrointestinal tract where absorption occurs (e.g., protein abundance in the crop, processing effects); and when appropriate, specific IgE binding studies or skin prick testing. Additional approaches may be considered (e.g., animal models; targeted sera screening) as the science evolves; however, such approaches have not been thoroughly evaluated or validated for predicting protein allergenicity. PMID: 18708115 [PubMed - indexed for MEDLINE] 98: Food Chem Toxicol. 2008 Oct;46 Suppl 10:S35-40. Epub 2008 Jul 30. Molecular profiles: a new tool to substantiate serum banks for evaluation of potential allergenicity of GMO. Barber D, Rodríguez R, Salcedo G. Departamento de I+D, ALK-Abelló S.A., C/Miguel Fleta 19, E-28037 Madrid, Spain. domingo.barber@alk-abello.com Assessment of the allergenicity of GMOs involves performing a test with a panel of sera obtained from allergic donors. However, there is no clear indication of how to characterize the above-mentioned panel. The patient selection criteria should take into account the geographical location of patients, the intensity and nature of the environmental allergens in the area and the potential cross-reactivity among allergenic molecules. Sera for serum banks, obtained from patients with demonstrated food allergy, should be subjected to a further characterization by screening with a panel of relevant allergenic molecules. A representative panel of these sera should be used in the allergenicity assessment. Finally, the "in vitro" methodologies should have the adequate specificity and sensitivity, and the integrity of the molecules tested should be guaranteed. Publication Types: Research Support, Non-U.S. Gov't PMID: 18706962 [PubMed - indexed for MEDLINE] 99: Food Chem Toxicol. 2008 Oct;46 Suppl 10:S12-4. Epub 2008 Jul 30. The serum bank of EuroPrevall - the prevalence, cost and basis of food allergy across Europe. Vieths S, Reese G, Ballmer-Weber BK, Beyer K, Burney P, Fernandez-Rivas M, Summers C, Ree R, Mills C. Paul-Ehrlich-Institut, Division of Allergology, Paul-Ehrlich-Strasse 51-59, D-63225, Langen, Germany. Viest@pei.de EuroPrevall is an EU-funded multidisciplinary project including 62 institutions from 22 countries. EuroPrevall studies the prevalence and distribution of food allergies in infants, children, adolescents, and adults in Europe, threshold doses for allergenic foods, the role of the environment in food allergy, the socioeconomic impact of food allergy, and novel diagnostic tools for food allergies. The EuroPrevall serum bank (EPASB), containing samples from approximately 70,000 subjects, is a major tool to achieve these goals. EPASB is coordinated by the Paul-Ehrlich-Institut, Langen, Germany. Local sera collections are administered at the University of Amsterdam (NL), the University Hospital of Manchester (UK), Charité Hospital (DE) and the Paul-Ehrlich-Institut. The EPASB coordinator and managing partners distribute samples for experimental work and regulate access. The overall aim is to provide sera to fulfil EuroPrevall research goals. The EPASB coordinator and managing partners suggest appropriate sera for addressing specific scientific and diagnostic questions. The serum bank will be maintained after termination of the project, but subsequent investigations must be in accordance with the original research goals of EuroPrevall. Thus, the contributors of the sera retain control over their future use. This rule prevents investigation of questions outside the scope of EuroPrevall, e.g. the allergenicity of genetically-modified foods. PMID: 18706961 [PubMed - indexed for MEDLINE] 100: Food Chem Toxicol. 2008 Oct;46 Suppl 10:S2-5. Epub 2008 Jul 30. Food allergy: a clinician's criteria for including sera in a serum bank. Ballmer-Weber BK, Fernández-Rivas M. Department of Dermatology, Allergy Unit, University Hospital Zürich, Gloriastrasse 31, CH 8091, Zürich, Switzerland. barbara.ballmer@usz.ch Safety assessment for genetically-engineered crop plants includes assessment for allergic responses. To facilitate this assessment, serum banks should contain well-characterised sera from patients with confirmed food allergies. A serum is defined as well-characterised if it is taken from a patient who has a convincing history of allergic responses to a known allergen or an allergen-containing food, a positive skin prick test (or elevated IgE response), and a positive response in a clinical food challenge. PMID: 18706467 [PubMed - indexed for MEDLINE] 101: Anal Bioanal Chem. 2008 Oct;392(3):333-40. Genetically modified food from crops: progress, pawns, and possibilities. Morin XK. Princeton Writing Program and the Princeton Environmental Institute, South Baker Hall, Whitman College, Princeton University, Princeton, NJ 08544, USA. xmorin@princeton.edu Publication Types: Research Support, Non-U.S. Gov't PMID: 18704376 [PubMed - indexed for MEDLINE] 102: Genes Brain Behav. 2008 Nov;7(8):924-32. Epub 2008 Aug 12. Construction of a taste-blind medaka fish and quantitative assay of its preference-aversion behavior. Aihara Y, Yasuoka A, Iwamoto S, Yoshida Y, Misaka T, Abe K. Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Tokyo, Japan. In vertebrates, the taste system provides information used in the regulation of food ingestion. In mammals, each cell group within the taste buds expresses either the T1R or the T2R taste receptor for preference-aversion discrimination. However, no such information is available regarding fish. We developed a novel system for quantitatively assaying taste preference-aversion in medaka fish. In this study, we prepared fluorescently labeled foods with fine cavities designed to retain tastants until they were bitten by the fish. The subjects were fed food containing a mixture of amino acids and inosine monophosphate (AN food), denatonium benzoate (DN food) or no tastant (NT food), and the amounts of ingested food were measured by fluorescence microscopy. Statistical analysis of the fluorescence intensities yielded quantitative measurements of AN food preference and DN food aversion. We then generated a transgenic fish expressing dominant-negative Galpha(i2) both in T1R-expressing and in T2R-expressing cells. The feeding assay revealed that the transgenic fish was unable to show a preference for AN food and an aversion to DN food. The assay system was useful for evaluating taste-blind behaviors, and the results indicate that the two taste signaling pathways conveying preferable and aversive taste information are conserved in fish as well as in mammals. Publication Types: Research Support, Non-U.S. Gov't PMID: 18700838 [PubMed - indexed for MEDLINE] 103: J Biol Chem. 2008 Nov 14;283(46):31871-83. Epub 2008 Aug 8. Centrins, cell cycle regulation proteins in human malaria parasite Plasmodium falciparum. Mahajan B, Selvapandiyan A, Gerald NJ, Majam V, Zheng H, Wickramarachchi T, Tiwari J, Fujioka H, Moch JK, Kumar N, Aravind L, Nakhasi HL, Kumar S. Division of Emerging and Transfusion Transmitted Diseases, Food and Drug Administration, Rockville, Maryland 20852, USA. Molecules and cellular mechanisms that regulate the process of cell division in malaria parasites remain poorly understood. In this study we isolate and characterize the four Plasmodium falciparum centrins (PfCENs) and, by growth complementation studies, provide evidence for their involvement in cell division. Centrins are cytoskeleton proteins with key roles in cell division, including centrosome duplication, and possess four Ca(2+)-binding EF hand domains. By means of phylogenetic analysis, we were able to decipher the evolutionary history of centrins in eukaryotes with particular emphasis on the situation in apicomplexans and other alveolates. Plasmodium possesses orthologs of four distinct centrin paralogs traceable to the ancestral alveolate, including two that are unique to alveolates. By real time PCR and/or immunofluorescence, we determined the expression of PfCEN mRNA or protein in sporozoites, asexual blood forms, gametocytes, and in the oocysts developing inside mosquito mid-gut. Immunoelectron microscopy studies showed that centrin is expressed in close proximity with the nucleus of sporozoites and asexual schizonts. Furthermore, confocal and widefield microscopy using the double staining with alpha-tubulin and centrin antibodies strongly suggested that centrin is associated with the parasite centrosome. Following the episomal expression of the four PfCENs in a centrin knock-out Leishmania donovani parasite line that exhibited a severe growth defect, one of the PfCENs was able to partially restore Leishmania growth rate and overcome the defect in cytokinesis in such mutant cell line. To our knowledge, this study is the first characterization of a Plasmodium molecule that is involved in the process of cell division. These results provide the opportunity to further explore the role of centrins in cell division in malaria parasites and suggest novel targets to construct genetically modified, live attenuated malaria vaccines. PMID: 18693242 [PubMed - indexed for MEDLINE] 104: Nat Biotechnol. 2008 Aug;26(8):858-9. Comment on: Nat Biotechnol. 2008 Feb;26(2):161-2. GMO testing-trade, labeling or safety first? Holst-Jensen A. Publication Types: Comment Letter PMID: 18688230 [PubMed - indexed for MEDLINE] 105: Natl Toxicol Program Genet Modif Model Rep. 2005 Oct;(1):1-222. NTP report on the toxicology studies of aspartame (CAS No. 22839-47-0) in genetically modified (FVB Tg.AC hemizygous) and B6.129-Cdkn2atm1Rdp (N2) deficient mice and carcinogenicity studies of aspartame in genetically modified [B6.129-Trp53tm1Brd (N5) haploinsufficient] mice (feed studies). Bucher JR, Bristol DW, French JE, Hailey JR, Haseman JK, Herbert RA, Malarkey DE, Maronpot RR, Peckham JC, Roycroft JH, Smith CS, Travlos GS, Vallant MK, Witt KL, Wenk ML, Lanning LL, Hardisty JF, Shackelford CC, Brecher S; NTP Pathology Working Group, Long P, Dixon D, Flake GP, Herbert RA, Little PB, Nyska A, Shackelford CC, Crockett PW, Betz LJ, McGowan KP, Scott JT, Gunnels SR, Carver PH, Coker KK, Hall BF, Harper LM, Rathman ES, Serbus DC, Willis RA. Aspartame is an artificial sweetener used throughout the world in food and beverages. Conventional 2-year rodent cancer studies of aspartame are considered negative, although a small number of neoplasms of the brain were observed in a rat study (Fed. Regist., 1981a,b). The NTP has explored the use of genetically altered mouse models as adjuncts to the 2-year rodent cancer assay. These models may prove to be more rapid, use fewer animals, and provide some mechanistic insights into neoplastic responses. As part of the evaluation of new mouse cancer screening models, aspartame was tested for potential toxicity and carcinogenicity in two relatively well-studied models, the Tg.AC hemizygous strain and the p53 haploinsufficient strain, and an uncharacterized model, the Cdkn2a deficient strain. Male and female Tg.AC hemizygous, p53 haploinsufficient, and Cdkn2a deficient mice were given feed containing aspartame (greater than 98% pure) for 9 months. Genetic toxicology studies were conducted in Salmonella typhimurium, rat bone marrow cells, and mouse peripheral blood erythrocytes. 9-MONTH STUDY IN Tg.AC HEMIZYGOUS MICE: Groups of 15 male and 15 female Tg.AC hemizygous mice were fed diets containing 0, 3,125, 6,250, 12,500, 25,000, or 50,000 ppm aspartame (equivalent to average daily doses of approximately 490, 980, 1,960, 3,960, or 7,660 mg aspartame/kg body weight to males and 550, 1,100, 2,260, 4,420, or 8,180 mg/kg to females) for 40 weeks. Exposure to aspartame had no effect on survival. The mean body weights of 50,000 ppm females were greater than those of the controls from week 15 until the end of the study. Feed consumption by the exposed groups was similar to that by the control groups throughout the study. There were no neoplasms or nonneoplastic lesions that were attributed to exposure to aspartame. 9-MONTH STUDY IN p53 HAPLOINSUFFICIENT MICE: Groups of 15 male and 15 female p53 haploinsufficient mice were fed diets containing 0, 3,125, 6,250, 12,500, 25,000, or 50,000 ppm aspartame (equivalent to average daily doses of approximately 490, 970, 1,860, 3,800, or 7,280 mg/kg to males and 630, 1,210, 2,490, 5,020, or 9,620 mg/kg to females) for 40 weeks. Exposure to aspartame had no effect on survival or mean body weights. Feed consumption by the exposed groups was similar to that by the control groups throughout the study. No neoplasms or nonneoplastic lesions were attributed to exposure to aspartame. 9-MONTH STUDY IN Cdkn2a DEFICIENT MICE: Groups of 15 male and 15 female Cdkn2a deficient mice were fed diets containing 0, 3,125, 6,250, 12,500, 25,000, or 50,000 ppm aspartame for 40 weeks (equivalent to average daily doses of approximately of approximately 490, 960, 1,900, 3,700, and 7,400 mg/kg to males and 610, 1,200, 2,390, 4,850, and 9,560 mg/kg to females). Survival of all exposed groups was similar to that of the control groups. Mean body weights of 3,125 and 6,250 ppm males were less than those of the controls after weeks 29 and 16, respectively. Mean body weights of female mice were similar to those of the controls throughout the study. The incidences of minimal to mild cytoplasmic vacuolization of periportal hepatocytes were significantly greater than controls in males exposed to 6,250, 25,000, or 50,000 ppm aspartame. GENETIC TOXICOLOGY: Aspartame was tested for induction of gene mutations in Salmonella typhimurium. No mutagenicity was detected in strains TA98, TA100, or TA1535 with or without exogenous metabolic activation (S9). In addition, a single test in TA1537 with 30% rat liver S9 gave negative results. In TA97 with 30% rat liver S9, however, a reproducible small increase in mutant colonies was observed, and this response was judged to be equivocal. No mutagenicity was detected in TA97 without S9 or with hamster liver S9. An acute bone marrow micronucleus test was conducted with aspartame administered by gavage to male F344/N rats. No increase in micronucleated polychromatic erythrocytes was observed at any dose level. Peripheral blood micronucleus tests were conducted after 9 months exposure of Tg.AC hemizygous, p53 haploinsufficient, and Cdkn2a deficient mice to aspartame in dosed feed. Negative results were obtained in male and female Tg.AC hemizygous and Cdkn2a deficient mice. Negative results were also obtained with male p53 haploinsufficient mice. In female p53 haploinsufficient mice, the results of the micronucleus test were judged to be positive, based on a significant trend test and a small but statistically significant increased frequency of micronucleated erythrocytes in the 50,000 ppm group. CONCLUSIONS Under the conditions of this 9-month feed study, there was no evidence of carcinogenic activity of aspartame in male or female p53 haploinsufficient mice exposed to 3,125, 6,250, 12,500, 25,000, or 50,000 ppm. Because this is a new model, there is uncertainty whether the study possessed sufficient sensitivity to detect a carcinogenic effect. PMID: 18685711 [PubMed - indexed for MEDLINE] 106: Biotechnol Bioeng. 2009 Jan 1;102(1):29-37. Cloning the bacterial bphC gene into Nicotiana tabacum to improve the efficiency of PCB phytoremediation. Novakova M, Mackova M, Chrastilova Z, Viktorova J, Szekeres M, Demnerova K, Macek T. ICT Prague, Faculty of Food and Biochemical Technology, Department of Biochemistry and Microbiology, Technicka 3, 16628 Prague, Czech Republic. The aim of this work is to increase the efficiency of the biodegradation of polychlorinated biphenyls (PCBs) by the introduction of bacterial genes into the plant genome. For this purpose, we selected the bphC gene encoding 2,3-dihydroxybiphenyl-1,2-dioxygenase from Pseudomonas testosteroni B-356 to be cloned into tobacco plants. The dihydroxybiphenyldioxygenase enzyme is the third enzyme in the biphenyl degradation pathway, and its unique function is the cleavage of biphenyl. Three different constructs were designed and prepared in E. coli: the bphC gene being fused with the beta-glucuronidase (GUS) gene, with the luciferase (LUC) gene, and with histidine tail in three separate plant cloning vectors. The GUS and LUC genes were chosen because they can be used as markers for the easy detection of transgenic plants, while histidine tail better enables the isolation of protein expressed in plant tissue. The prepared vectors were then introduced into cells of Agrobacterium tumefaciens. The transient expression of the prepared genes was first studied in cells of Nicotiana tabacum. Once this ability had been established, model tobacco plants were transformed by agrobacterial infection with the bphC/GUS, bphC/LUC, and bphC/His genes. The transformed regenerants were selected on media using a selective antibiotic, and the presence of transgenes and mRNA was determined by PCR and RT-PCR. The expression of the fused proteins BphC/GUS and BphC/LUC was confirmed histochemically by analysis of the expression of their detection markers. Western blot analysis was performed to detect the presence of the BphC/His protein immunochemically using a mouse anti-His antibody. Growth and viability of transgenic plants in the presence of PCBs was compared with control plants. Publication Types: Research Support, Non-U.S. Gov't PMID: 18683252 [PubMed - indexed for MEDLINE] 107: Dev Biol. 2008 Oct 1;322(1):46-55. Epub 2008 Jul 9. Sensory mechanisms controlling the timing of larval developmental and behavioral transitions require the Drosophila DEG/ENaC subunit, Pickpocket1. Ainsley JA, Kim MJ, Wegman LJ, Pettus JM, Johnson WA. University of Iowa, Roy J. and Lucille A. Carver College of Medicine, Department of Molecular Physiology and Biophysics, Iowa City, IA 52242, USA. Growth of multicellular organisms proceeds through a series of precisely timed developmental events requiring coordination between gene expression, behavioral changes, and environmental conditions. In Drosophila melanogaster larvae, the essential midthird instar transition from foraging (feeding) to wandering (non-feeding) behavior occurs prior to pupariation and metamorphosis. The timing of this key transition is coordinated with larval growth and size, but physiological mechanisms regulating this process are poorly understood. Results presented here show that Drosophila larvae associate specific environmental conditions, such as temperature, with food in order to enact appropriate foraging strategies. The transition from foraging to wandering behavior is associated with a striking reversal in the behavioral responses to food-associated stimuli that begins early in the third instar, well before food exit. Genetic manipulations disrupting expression of the Degenerin/Epithelial Sodium Channel subunit, Pickpocket1(PPK1) or function of PPK1 peripheral sensory neurons caused defects in the timing of these behavioral transitions. Transient inactivation experiments demonstrated that sensory input from PPK1 neurons is required during a critical period early in the third instar to influence this developmental transition. Results demonstrate a key role for the PPK1 sensory neurons in regulation of important behavioral transitions associated with developmental progression of larvae from foraging to wandering stage. Publication Types: Research Support, Non-U.S. Gov't PMID: 18674528 [PubMed - indexed for MEDLINE] 108: Vopr Pitan. 2008;77(3):58-63. [Questions safety and tendency of using genetically modified microorganisms in food, food additives and food derived] [Article in Russian] Khovaev AA. In this article analysis questions of using genetically modified microorganisms in manufacture food production, present new GMM used in manufacture -food ferments; results of medical biological appraisal/ microbiological and genetic expert examination/ of food, getting by use microorganisms or there producents with indication modern of control methods. Publication Types: English Abstract Review PMID: 18669333 [PubMed - indexed for MEDLINE] 109: Vopr Pitan. 2008;77(3):49-57. [Requirements to a medical and biologic assessment and the hygienic control of the food production received from recombinant-DNA microorganisms] [Article in Russian] Sheveleva SA, Efimmochkina NR, Nesterenko LN, Zigangirova NA, Khovaev AA, Naroditskiĭ BS, Ivanov GE, Tutel'ian VA, Gintsburg AL. In work the characteristic of the created in the Russian Federation system of an estimation of safety of the foodstuff received from/or with use of genetically modified microorganisms (GMM) is given, at their admission to realization and the hygienic control of given production over a revolution. It is shown, that strategy of a safety at a stage of registration GMM, the established order and accepted control measures of the foodstuff received from/or with use GMM, in Russia their large-scale commercial use, and the normative-legal and methodical base based on the federal legislation on state regulation in the field of genetically engineering activity, about quality and effectively outstrip safety of foodstuff about protection of the rights of consumers, is harmonized with approaches of the international organizations. Publication Types: English Abstract Review PMID: 18669332 [PubMed - indexed for MEDLINE] 110: Anal Bioanal Chem. 2008 Oct;392(3):395-404. Epub 2008 Jul 31. Real-time and conventional PCR detection of Liberty Link rice varieties and transgenic soy in rice sampled in the Mexican and American retail markets. Quirasco M, Schoel B, Chhalliyil P, Fagan J, Gálvez A. Departamento de Alimentos y Biotecnología, Facultad de Química, Universidad Nacional Autónoma de México, 04510, México, D. F., México. Samples of rice from Mexican and USA retail stores were analyzed for the presence of transgenic (GM) events using real-time PCR. In screening for the CaMV35S promoter sequence (35SP), positive results were found in 49 and 35% of the Mexican and American samples, respectively. In further investigations in Mexican samples, 43% were positive for P35S::bar, with two above the quantifiable limit; these were 0.07% and 0.05% GMO. Fourteen out of the sixteen positive samples were labeled as imported from the USA. In testing samples bought in American retail shops, 24% showed positive results, all below the quantifiable range. It could be deduced that P35S::bar positive samples were Liberty Link(R) (LL) rice. In distinguishing between LL601 and LL62, end-point PCR was used, corroborating the P35S::bar amplicon length difference of these events. LL62 was found in one rice sample purchased in Mexico and two in the USA samples. Its presence was verified with the 35S terminator sequence. All other LL positive samples contained LL601. None of the samples analyzed showed the presence of Bt63 rice. The LL rice varieties found have been identified as not being commercially cultivated, and so their presence requires further investigation. 35SP was also present in samples which did not have any LL rice. Maize sequences could not be detected in any of the samples; however, soybean DNA was found in Mexican and USA rice samples. The Roundup Ready(R) trait was detected in trace amounts in 16 and 6% of the rice samples bought in Mexico and the USA, respectively. Real-time PCR was shown to be the method of choice for the sensitive and rapid screening of commodities and retail samples for the detection of GM and other contamination. PMID: 18668229 [PubMed - indexed for MEDLINE] 111: Horm Behav. 2008 Sep;54(4):506-13. Epub 2008 Mar 10. Disruption of seasonality in growth hormone-transgenic coho salmon (Oncorhynchus kisutch) and the role of cholecystokinin in seasonal feeding behavior. Lõhmus M, Raven PA, Sundström LF, Devlin RH. Fisheries and Oceans Canada, Centre for Aquaculture and Environmental Research, 4160 Marine Drive, West Vancouver, V7V 1N6, BC Canada. Seasonal variation in daily food intake is a well-documented phenomenon in many organisms including wild-type coho salmon where the appetite is noticeably reduced during periods of decreased day length and low water temperature. This reduction may in part be explained by altered production of cholecystokinin (CCK) and growth hormone (GH). CCK is a hormone produced in the brain and gut that mediates a feeling of satiety and thus has an inhibitory effect on food intake and foraging behaviour. Growth hormone (GH) enhances feeding behaviour and consequently growth, but its production is reduced during winter. The objectives of this study were: first, to compare the seasonal feeding behaviour of wild and GH-transgenic coho salmon; second, to determine the behavioural effect of blocking the action of CCK (by using devazepide) on the seasonal food intake; and third, to measure CCK expression in brain and gut tissues between the two genotypes across seasons. We found that, in contrast to wild salmon, food intake in transgenic salmon was not reduced during winter indicating that seasonal control of appetite regulation has been disrupted by constitutive production of GH in transgenic animals. Blocking of CCK increased food intake in both genotypes in all seasons. The increase was stronger in wild genotypes than transgenic fish; however blocking CCK in wild-type fish in winter did not elevate appetites to levels observed in the summer. The response to devazepide was generally faster in transgenic than in wild salmon with more rapid effects observed during summer than during winter, possibly due to a higher temperature in summer. Overall, a seasonal effect on CCK mRNA levels was observed in telencephalon with levels during winter being higher compared to the summer in wild fish, but with no seasonal effect in transgenic fish. No differences in seasonal CCK expression were found in hypothalamus. Higher levels of CCK were detected in the gut of both genotypes in winter compared to summer. Thus, CCK appears to mediate food intake among seasons in both wild-type and GH-transgenic salmon, and an altered CCK regulation may be responsible at least in part for the seasonal regulation of food intake. Publication Types: Comparative Study Research Support, Non-U.S. Gov't PMID: 18667200 [PubMed - indexed for MEDLINE] 112: Transgenic Res. 2008 Dec;17(6):1025-33. Epub 2008 Jul 29. Animal pharming, two decades on. Kind A, Schnieke A. Livestock Biotechnology, Technische Universität München, Center of Life and Food Sciences Weihenstephan, Hochfeldweg, 1, Freising 85354, Germany. Since its inception 20 years ago, the animal pharming industry has promoted transgenic animals as a cost-effective method of biopharmaceutical production. However, it took until 2006 for the first therapeutic product to gain regulatory approval. This was an important milestone, but scepticism still abounds. Can pharming regain investor confidence, and will society accept transgenic livestock as a production method? There is some cause for optimism, biopharmaceuticals are a large, expanding market and animal pharming has already made considerable strides. A novel production platform has been established, groundbreaking technologies developed, a necessary regulatory framework put in place. Nevertheless, despite cost advantages, pharming has become a niche production method and its long term success may depend on products unique to transgenic animals. Publication Types: Review PMID: 18663595 [PubMed - indexed for MEDLINE] 113: Plant Biotechnol J. 2008 Oct;6(8):843-53. Epub 2008 Jul 23. Erratum in: Plant Biotechnol J. 2008 Oct;6(8):854. Low-acrylamide French fries and potato chips. Rommens CM, Yan H, Swords K, Richael C, Ye J. Simplot Plant Sciences, JR Simplot Company, Boise, ID 83706, USA. crommens@simplot.com SUMMARY: Acrylamide is produced in starchy foods that are baked, roasted or fried at high temperatures. Concerns about the potential health issues associated with the dietary intake of this reactive compound led us to reduce the accumulation of asparagine, one of its main precursors, in the tubers of potato (Solanum tuberosum). This metabolic change was accomplished by silencing two asparagine synthetase genes through 'all-native DNA' transformation. Glasshouse-grown tubers of the transformed intragenic plants contained up to 20-fold reduced levels of free asparagine. This metabolic change coincided with a small increase in the formation of glutamine and did not affect tuber shape or yield. Heat-processed products derived from the low-asparagine tubers were also indistinguishable from their untransformed counterparts in terms of sensory characteristics. However, both French fries and potato chips accumulated as little as 5% of the acrylamide present in wild-type controls. Given the important role of processed potato products in the modern Western diet, a replacement of current varieties with intragenic potatoes could reduce the average daily intake of acrylamide by almost one-third. PMID: 18662372 [PubMed - indexed for MEDLINE] 114: Genome. 2008 Aug;51(8):668-72. foxo is required for resistance to amino acid starvation in Drosophila. Kramer JM, Slade JD, Staveley BE. Department of Biology, Memorial University of Newfoundland, St. John's, NL A1B3X9, Canada. bestave@mun.ca The ability of an organism to alter its metabolism, growth, and reproductive capacity in response to fluctuations in food availability has likely been an important factor in the course of evolution. The insulin signalling pathway is an evolutionarily conserved mechanism used by metazoan animals to sense and respond to changes in nutrient intake. During conditions of starvation the level of circulating insulin is low. Under conditions of low insulin, the foxo family of transcription factors are activated. Studies in Drosophila suggest that Drosophila foxo may alter the transcriptional profile of cells to allow for maximum survival of the fly during starvation. We have tested this ability in transgenic flies containing a luciferase reporter gene under the control of foxo response elements. We show that foxo activity is increased during amino acid starvation and reduced in the presence of amino acids. In addition, we find that loss of function of foxo leads to reduced survival under conditions of amino acid starvation in both larvae and adult flies. These data provide direct evidence that foxo is activated during amino acid starvation and is critical for optimal survival under these conditions. Publication Types: Research Support, Non-U.S. Gov't PMID: 18650956 [PubMed - indexed for MEDLINE] 115: Chem Biodivers. 2008 Jul;5(7):1225-37. Synthetic antimicrobial peptides as agricultural pesticides for plant-disease control. Montesinos E, Bardají E. Institute of Food and Agricultural Technology-CIDSAV-XaRTA, University of Girona, Campus Montilivi, E-18071 Girona. emonte@intea.udg.edu There is a need of antimicrobial compounds in agriculture for plant-disease control, with low toxicity and reduced negative environmental impact. Antimicrobial peptides are produced by living organisms and offer strong possibilities in agriculture because new compounds can be developed based on natural structures with improved properties of activity, specificity, biodegradability, and toxicity. Design of new molecules has been achieved using combinatorial-chemistry procedures coupled to high-throughput screening systems and data processing with design-of-experiments (DOE) methodology to obtain QSAR equation models and optimized compounds. Upon selection of best candidates with low cytotoxicity and moderate stability to protease digestion, anti-infective activity has been evaluated in plant-pathogen model systems. Suitable compounds have been submitted to acute toxicity testing in higher organisms and exhibited a low toxicity profile in a mouse model. Large-scale production can be achieved by solution organic or chemoenzymatic procedures in the case of very small peptides, but, in many cases, production can be performed by biotechnological methods using genetically modified microorganisms (fermentation) or transgenic crops (plant biofactories). Publication Types: Review PMID: 18649311 [PubMed - indexed for MEDLINE] 116: Histochem Cell Biol. 2008 Nov;130(5):967-77. Epub 2008 Jul 22. A long-term study on female mice fed on a genetically modified soybean: effects on liver ageing. Malatesta M, Boraldi F, Annovi G, Baldelli B, Battistelli S, Biggiogera M, Quaglino D. Dipartimento di Scienze Morfologico-Biomediche, Sezione di Anatomia e Istologia, University of Verona, strada Le Grazie 8, 37134, Verona, Italy. manuela.malatesta@univr.it Liver represents a suitable model for monitoring the effects of a diet, due to its key role in controlling the whole metabolism. Although no direct evidence has been reported so far that genetically modified (GM) food may affect health, previous studies on hepatocytes from young female mice fed on GM soybean demonstrated nuclear modifications involving transcription and splicing pathways. In this study, the effects of this diet were studied on liver of old female mice in order to elucidate possible interference with ageing. The morpho-functional characteristics of the liver of 24-month-old mice, fed from weaning on control or GM soybean, were investigated by combining a proteomic approach with ultrastructural, morphometrical and immunoelectron microscopical analyses. Several proteins belonging to hepatocyte metabolism, stress response, calcium signalling and mitochondria were differentially expressed in GM-fed mice, indicating a more marked expression of senescence markers in comparison to controls. Moreover, hepatocytes of GM-fed mice showed mitochondrial and nuclear modifications indicative of reduced metabolic rate. This study demonstrates that GM soybean intake can influence some liver features during ageing and, although the mechanisms remain unknown, underlines the importance to investigate the long-term consequences of GM-diets and the potential synergistic effects with ageing, xenobiotics and/or stress conditions. PMID: 18648843 [PubMed - indexed for MEDLINE] 117: Plant Cell Environ. 2008 Oct;31(10):1410-5. Epub 2008 Jul 14. Isoprene emissions influence herbivore feeding decisions. Laothawornkitkul J, Paul ND, Vickers CE, Possell M, Taylor JE, Mullineaux PM, Hewitt CN. Lancaster Environment Centre, Lancaster University, Lancaster, UK. Isoprene (C(5)H(8), 2-methyl 1,3-butadiene) is synthesized and emitted by many, but not all, plants. Unlike other related volatile organic compounds (monoterpenes and sesquiterpenes), isoprene has not been shown to mediate plant-herbivore interactions. Here, for the first time, we show, in feeding choice tests using isoprene-emitting transgenic tobacco plants (Nicotiana tabacum cv. Samsun) and non-emitting azygous control plants, that isoprene deters Manduca sexta caterpillars from feeding. This avoidance behaviour was confirmed using an artificial (isoprene-emitting and non-emitting control) diet. Both in vivo and in vitro experiments showed that isoprene can activate feeding avoidance behaviour in this system with a dose-response effect on caterpillar behaviour and an isoprene emission threshold level of <6 nmol m(-2) s(-1). Publication Types: Research Support, Non-U.S. Gov't PMID: 18643955 [PubMed - indexed for MEDLINE] 118: Physiol Plant. 2008 Nov;134(3):394-402. Epub 2008 Jul 31. Overexpression of the apple alcohol acyltransferase gene alters the profile of volatile blends in transgenic tobacco leaves. Li D, Shen J, Wu T, Xu Y, Zong X, Li D, Shu H. College of Food Sciences, Shandong Agricultural University, Tai'an, Shandong 271018, P.R. China. Alcohol acyltransferases (AATs) are key enzymes in ester biosynthesis. Previous studies have found that AAT may be a stress-related gene. To investigate further the function of the apple alcohol acyltransferase gene (MdAAT2), transgenic tobacco plants overexpressing MdAAT2 were generated. Gas chromatography-mass spectroscopy analysis showed that the volatile blends were altered in these transgenic tobacco leaves. Although no apple-fruity volatile esters were detected in transgenic tobacco leaves, methyl caprylate, methyl caprate, and methyl dodecanoate were newly generated, and the concentrations of methyl benzoate and methyl tetradecanoate were significantly increased, suggesting that MdAAT2 may use medium-chain fatty acyl CoA and benzoyl-CoA as acyl donors together with methanol acceptors as substrates. Surprisingly, the concentrations of linalool were significantly increased in transgenic tobacco leaves, which may mediate the repellent effect on Myzus persicae (Sulzer) aphids. Using methyl jasmonate (MeJA) and wounding treatments, we found that MdAAT2 may substitute for the partial ability of MeJA to induce the production of linalool in transgenic plants. These data suggest that MdAAT2 may be involved in the response to the MeJA signal and may play a role in the response to biotic and abiotic stress. Publication Types: Research Support, Non-U.S. Gov't PMID: 18636987 [PubMed - indexed for MEDLINE] 119: J Agric Food Chem. 2008 Aug 27;56(16):6791-800. Epub 2008 Jul 16. Detection of genetically modified canola using multiplex PCR coupled with oligonucleotide microarray hybridization. Schmidt AM, Sahota R, Pope DS, Lawrence TS, Belton MP, Rott ME. Sidney Laboratory, Canadian Food Inspection Agency, 8801 East Saanich Road, Sidney, British Columbia V8L 1H3, Canada. A rapid method was developed for concurrent screening of transgenic elements in GM canola. This method utilizes a single multiplex PCR coupled with an oligonucleotide DNA array capable of simultaneously detecting the 12 approved GM canola lines in Canada. The assay includes construct-specific elements for identification of approved lines, common elements (e.g., CaMV 35S promoter, Agrobacterium tumefaciens nos terminator, or nptII gene) for screening of approved or unapproved lines, a canola-specific endogenous gene, and endogenous genes from heterologous crops to serve as additional controls. Oligonucleotide probes were validated individually for functionality and specificity by amplification of specific transgene sequences from appropriate GM canola lines corresponding to each probe sequence, and hybridization of amplicons to the array. Each target sequence hybridized to its corresponding oligonucleotide probe and no significant cross-hybridization was observed. The limit of detection was examined for the GM lines GT73, T45, and MS8/RF3, and was determined to be 0.1%, 0.1%, and 0.5%, respectively, well within the European food and feed labeling threshold level of 0.9% for approved GM product. Practically, the method was demonstrated to be effective for the detection of GM canola in several types of animal feed, as well as in commercial canola meal. PMID: 18636685 [PubMed - indexed for MEDLINE] 120: J Biotechnol. 2008 Sep 10;136(3-4):140-7. Epub 2008 May 28. Hydrolysis of amorphous and crystalline cellulose by heterologously produced cellulases of Melanocarpus albomyces. Szijártó N, Siika-Aho M, Tenkanen M, Alapuranen M, Vehmaanperä J, Réczey K, Viikari L. Department of Applied Biotechnology and Food Science, Budapest University of Technology and Economics, P.O. Box 91, H-1521 Budapest, Hungary. nora_szijarto@mkt.bme.hu Three thermostable neutral cellulases from Melanocarpus albomyces, a 20-kDa endoglucanase (Cel45A), a 50-kDa endoglucanase (Cel7A), and a 50-kDa cellobiohydrolase (Cel7B) heterologously produced in a recombinant Trichoderma reesei were purified and studied in hydrolysis (50 degrees C, pH 6.0) of crystalline and amorphous cellulose. To improve their efficiency, M. albomyces cellulases naturally harboring no cellulose-binding module (CBM) were genetically modified to carry the CBM of T. reesei CBHI/Cel7A, and were studied under similar experimental conditions. Hydrolysis performance and product profiles were used to evaluate hydrolytic features of the investigated enzymes. Each cellulase proved to be active against the tested substrates; the cellobiohydrolase Cel7B had greater activity than the endoglucanases Cel45A and Cel7A against crystalline cellulose, whereas in the case of amorphous substrate the order was reversed. Evidence of synergism was observed when mixtures of the novel enzymes were applied in a constant total protein dosage. Presence of the CBM improved the hydrolytic potential of each enzyme in all experimental configurations; it had a greater effect on the endoglucanases Cel45A and Cel7A than the cellobiohydrolase Cel7B, especially against crystalline substrate. The novel cellobiohydrolase performed comparably to the major cellobiohydrolase of T. reesei (CBHI/Cel7A) under the applied experimental conditions. Publication Types: Research Support, Non-U.S. Gov't PMID: 18635283 [PubMed - indexed for MEDLINE] 121: Plant Physiol. 2008 Sep;148(1):89-96. Epub 2008 Jul 16. Expression of Umbelopsis ramanniana DGAT2A in seed increases oil in soybean. Lardizabal K, Effertz R, Levering C, Mai J, Pedroso MC, Jury T, Aasen E, Gruys K, Bennett K. Calgene Campus, Monsanto Company, Davis, California 95616, USA. kathy.lardizabal@monsanto.com Oilseeds are the main source of lipids used in both food and biofuels. The growing demand for vegetable oil has focused research toward increasing the amount of this valuable component in oilseed crops. Globally, soybean (Glycine max) is one of the most important oilseed crops grown, contributing about 30% of the vegetable oil used for food, feed, and industrial applications. Breeding efforts in soy have shown that multiple loci contribute to the final content of oil and protein stored in seeds. Genetically, the levels of these two storage products appear to be inversely correlated with an increase in oil coming at the expense of protein and vice versa. One way to overcome the linkage between oil and protein is to introduce a transgene that can specifically modulate one pathway without disrupting the other. We describe the first, to our knowledge, transgenic soy crop with increased oil that shows no major impact on protein content or yield. This was achieved by expressing a codon-optimized version of a diacylglycerol acyltransferase 2A from the soil fungus Umbelopsis (formerly Mortierella) ramanniana in soybean seed during development, resulting in an absolute increase in oil of 1.5% (by weight) in the mature seed. Publication Types: Research Support, Non-U.S. Gov't PMID: 18633120 [PubMed - indexed for MEDLINE] 122: PLoS ONE. 2008 Jul 16;3(7):e2664. Potential effects of oilseed rape expressing oryzacystatin-1 (OC-1) and of purified insecticidal proteins on larvae of the solitary bee Osmia bicornis. Konrad R, Ferry N, Gatehouse AM, Babendreier D. Agroscope Reckenholz-Tänikon Research Station ART, Zürich, Switzerland. roger.konrad@art.admin.ch Despite their importance as pollinators in crops and wild plants, solitary bees have not previously been included in non-target testing of insect-resistant transgenic crop plants. Larvae of many solitary bees feed almost exclusively on pollen and thus could be highly exposed to transgene products expressed in the pollen. The potential effects of pollen from oilseed rape expressing the cysteine protease inhibitor oryzacystatin-1 (OC-1) were investigated on larvae of the solitary bee Osmia bicornis (= O. rufa). Furthermore, recombinant OC-1 (rOC-1), the Bt toxin Cry1Ab and the snowdrop lectin Galanthus nivalis agglutinin (GNA) were evaluated for effects on the life history parameters of this important pollinator. Pollen provisions from transgenic OC-1 oilseed rape did not affect overall development. Similarly, high doses of rOC-1 and Cry1Ab as well as a low dose of GNA failed to cause any significant effects. However, a high dose of GNA (0.1%) in the larval diet resulted in significantly increased development time and reduced efficiency in conversion of pollen food into larval body weight. Our results suggest that OC-1 and Cry1Ab expressing transgenic crops would pose a negligible risk for O. bicornis larvae, whereas GNA expressing plants could cause detrimental effects, but only if bees were exposed to high levels of the protein. The described bioassay with bee brood is not only suitable for early tier non-target tests of transgenic plants, but also has broader applicability to other crop protection products. Publication Types: Research Support, Non-U.S. Gov't PMID: 18628826 [PubMed - indexed for MEDLINE] 123: Mol Plant Microbe Interact. 2008 Jun;21(6):757-68. WRR4 encodes a TIR-NB-LRR protein that confers broad-spectrum white rust resistance in Arabidopsis thaliana to four physiological races of Albugo candida. Borhan MH, Gunn N, Cooper A, Gulden S, Tör M, Rimmer SR, Holub EB. Agriculture and Agri-Food Canada, Saskatoon Research Centre, Saskatoon, SK., S7N 0X2, Canada. White blister rust in the Brassicaceae is emerging as a superb model for exploring how plant biodiversity has channeled speciation of biotrophic parasites. The causal agents of white rust across a wide breadth of cruciferous hosts currently are named as variants of a single oomycete species, Albugo candida. The most notable examples include a major group of physiological races that each are economically destructive in a different vegetable or oilseed crop of Brassica juncea (A. candida race 2), B. rapa (race 7), or B. oleracea (race 9); or parasitic on wild crucifers such as Capsella bursa-pastoris (race 4). Arabidopsis thaliana is innately immune to these races of A. candida under natural conditions; however, it commonly hosts its own molecularly distinct subspecies of A. candida (A. candida subsp. arabidopsis). In the laboratory, we have identified several accessions of Arabidopsis thaliana (e.g.,. Ws-3) that can permit varying degrees of rust development following inoculation with A. candida races 2, 4, and 7, whereas race 9 is universally incompatible in Arabidopsis thaliana and nonrusting resistance is the most prevalent outcome of interactions with the other races. Subtle variation in resistance phenotypes is evident, observed initially with an isolate of A. candida race 4, indicating additional genetic variation. Therefore, we used the race 4 isolate for map-based cloning of the first of many expected white rust resistance (WRR) genes. This gene was designated WRR4 and encodes a cytoplasmic toll-interleukin receptor-like nucleotide-binding leucine-rich repeat receptor-like protein that confers a dominant, broad-spectrum white rust resistance in the Arabidopsis thaliana accession Columbia to representative isolates of A. candida races 2, 4, 7, and 9, as verified by transgenic expression of the Columbia allele in Ws-3. The WRR4 protein requires functional expression of the lipase-like protein EDS1 but not the paralogous protein PAD4, and confers full immunity that masks an underlying nonhypersensitive incompatibility in Columbia to A. candida race 4. This residual incompatibility is independent of functional EDS1. Publication Types: Research Support, Non-U.S. Gov't PMID: 18624640 [PubMed - indexed for MEDLINE] 124: Science. 2008 Jul 11;321(5886):184-5. Food safety. Arsenic and paddy rice: a neglected cancer risk? Stone R. Publication Types: News PMID: 18621644 [PubMed - indexed for MEDLINE] 125: Plant Cell Rep. 2008 Sep;27(9):1423-40. Epub 2008 Jul 9. A history of plant biotechnology: from the Cell Theory of Schleiden and Schwann to biotech crops. Vasil IK. University of Florida, Box 110690, Gainesville, FL 32611-0690, USA. ivasil@ufl.edu Plant biotechnology is founded on the principles of cellular totipotency and genetic transformation, which can be traced back to the Cell Theory of Matthias Jakob Schleiden and Theodor Schwann, and the discovery of genetic transformation in bacteria by Frederick Griffith, respectively. On the 25th anniversary of the genetic transformation of plants, this review provides a historical account of the evolution of the theoretical concepts and experimental strategies that led to the production and commercialization of biotech (transformed or transgenic) plants expressing many useful genes, and emphasizes the beneficial effects of plant biotechnology on food security, human health, the environment, and conservation of biodiversity. In so doing, it celebrates and pays tribute to the contributions of scores of scientists who laid the foundation of modern plant biotechnology by their bold and unconventional thinking and experimentation. It highlights also the many important lessons to be learnt from the fascinating history of plant biotechnology, the significance of history in science teaching and research, and warns against the danger of the growing trends of ignoring history and historical illiteracy. Publication Types: Historical Article Review PMID: 18612644 [PubMed - indexed for MEDLINE] 126: Plant Physiol. 2008 Jul;147(3):939-53. Nutritionally improved agricultural crops. Newell-McGloughlin M. University of California, Systemwide Biotechnology Research and Education Program, Davis, California 95616, USA. mmmcgloughlin@ucdavis.edu PMID: 18612071 [PubMed - indexed for MEDLINE] 127: Regul Toxicol Pharmacol. 2008 Nov;52(2):94-103. Epub 2008 Jun 20. Establishing objective detection limits for the pepsin digestion assay used in the assessment of genetically modified foods. Ofori-Anti AO, Ariyarathna H, Chen L, Lee HL, Pramod SN, Goodman RE. Food Allergy Research and Resource Program (FARRP), Department of Food Science & Technology, University of Nebraska-Lincoln, 143 Food Industry Complex, Lincoln, NE 68583-0955, USA. RATIONALE: Guidelines for assessing the potential allergenicity of genetically modified (GM) organisms recommend testing the digestibility of the introduced protein by pepsin. Previous studies detailed the digestion procedure but have not described a simple objective measurement of the extent of digestion nor evaluated the impact of variation in pepsin activity. METHODS: Samples of eight proteins were digested by pepsin at pH 1.2 and 2.0 using standard conditions (10,000 U of pepsin activity per mg test protein) as well as 5000 and 20,000 units per mg of test protein. An independent digestion assay of hemoglobin was used to verify pepsin activity for each assay. Digestion was stopped in timed samples between 0.5 and 60 min. Digestion samples and undigested protein (10% and 100%) were separated by SDS-PAGE. Residual stained protein bands were measured by image analysis. RESULTS: The differences in pH and pepsin concentration only had minor effects on digestion of intermediately stable proteins: concanavalin A, ovalbumin, and lysozyme, but not on rapidly digested or stable proteins. CONCLUSIONS: Verification of pepsin activity and measurement of an objective endpoint of digestion (e.g. (90%) should provide more comparable results for the safety assessment of novel food proteins. Publication Types: Research Support, Non-U.S. Gov't PMID: 18611423 [PubMed - indexed for MEDLINE] 128: Biotechnol Annu Rev. 2008;14:423-62. Recent advances in the development of transgenic papaya technology. Tecson Mendoza EM, C Laurena A, Botella JR. College of Agriculture, University of the Philippines Los Baños, College, Laguna, Philippines. emtmendoza@nast.ph Papaya with resistance to papaya ringspot virus (PRSV) is the first genetically modified tree and fruit crop and also the first transgenic crop developed by a public institution that has been commercialized. This chapter reviews the different transformation systems used for papaya and recent advances in the use of transgenic technology to introduce important quality and horticultural traits in papaya. These include the development of the following traits in papaya: resistance to PRSV, mites and Phytophthora, delayed ripening trait or long shelf life by inhibiting ethylene production or reducing loss of firmness, and tolerance or resistance to herbicide and aluminum toxicity. The use of papaya to produce vaccine against tuberculosis and cysticercosis, an infectious animal disease, has also been explored. Because of the economic importance of papaya, there are several collaborative and independent efforts to develop PRSV transgenic papaya technology in 14 countries. This chapter further reviews the strategies and constraints in the adoption of the technology and biosafety to the environment and food safety. Constraints to adoption include public perception, strict and expensive regulatory procedures and intellectual property issues. Publication Types: Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Review PMID: 18606373 [PubMed - indexed for MEDLINE] 129: Biotechnol Annu Rev. 2008;14:171-90. Use of the cauliflower Or gene for improving crop nutritional quality. Zhou X, Van Eck J, Li L. U.S. Department of Agriculture- Department of Plant Breeding and Genetics, Cornell University, Ithaca, NY 14853, USA. Carotenoids are a group of pigments that are essential to human diets. An increasing interest in carotenoids as a nutritional source of vitamin A and health-promoting compounds has prompted the recent progress in metabolic engineering of carotenogenesis in food crops. Current strategies have been mainly focused on manipulating genes encoding carotenogeic enzymes. In many cases, it is difficult to reach the desired levels of carotenoid enhancement. In this chapter, we briefly summarize the recent progress on our understanding of carotenoid biosynthesis. We describe the isolation of a novel gene, the Or gene, from a high-beta-carotene orange cauliflower mutant. The Or gene encodes a plastid-targeted protein containing a cysteine-rich zinc finger domain and appears to be plant-specific. The insertion of a copia-like LTR retrotransponson in the Or gene confers high levels of carotenoid accumulation in the normally low-pigmented tissues. Rather than directly regulating carotenoid biosynthesis, the Or gene controls carotenoid accumulation by inducing the formation of chromoplasts, which provide a metabolic sink to sequester and deposit carotenoids. Examination of the Or transgenic potato tubers confirms that the Or-induced carotenoid accumulation is associated with the formation of a metabolic sink. Thus, the Or gene offers a new molecular tool to complement current approaches for nutritional enhancement in agriculturally important crops. Publication Types: Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Review PMID: 18606363 [PubMed - indexed for MEDLINE] 130: Plant Physiol Biochem. 2008 Oct;46(10):868-74. Epub 2008 Jun 3. A red algal cyclophilin has an effect on development and growth in Nicotiana tabacum. Cho EK, Kim M. Department of Bio-Food Materials, College of Medical Life Science, Silla University, Busan, Republic of Korea. choeun@silla.ac.kr In this study, an algal Cyp was introduced into plant to research the effect of the gene on growth and development. cDNA GjCyp-1 was isolated from the red alga (Griffithsia japonica), and a recombinant GjCyp-1 containing a CaMV35S promoter at the amino-terminus was constructed in Nicotiana tabacum. The altered GjCyp-1 levels in plants and the expression pattern of Cyp after hormone treatment were confirmed by RNA blotting. Transcript of GjCyp-1 was induced by plant hormones such as gibberellic acid (GA(3)), indoleacetic acid (IAA), and zeatin (ZA). Constitutive overexpression of GjCyp-1 appeared to be beneficial to seed germination. The ratio of emergence of cotyledon from seeds overexpressing GjCyp-1 was almost three times higher than that of the transgenic seeds carrying only the vector. In addition, it was found that most of the seedlings overexpressing GjCyp-1 were dwarfs with altered root systems. The ratio of leaf length and width and root length from transgenic seedlings overexpressing GjCyp-1 was almost 2 and 3.5 times lower than that of the transgenic seedlings carrying only the vector, respectively. The data in this study suggest that GjCyp-1 may affect development and growth in organisms. PMID: 18603440 [PubMed - indexed for MEDLINE] 131: Food Chem Toxicol. 2008 Aug;46(8):2591-605. Epub 2008 Jun 3. Safety assessment considerations for food and feed derived from plants with genetic modifications that modulate endogenous gene expression and pathways. Kier LD, Petrick JS. Monsanto Company, 800 North Lindbergh Blvd., Mail Code O3F, St. Louis, MO 63167, USA. The current globally recognized comparative food and feed safety assessment paradigm for biotechnology-derived crops is a robust and comprehensive approach for evaluating the safety of both the inserted gene product and the resulting crop. Incorporating many basic concepts from food safety, toxicology, nutrition, molecular biology, and plant breeding, this approach has been used effectively by scientists and regulatory agencies for 10-15 years. Current and future challenges in agriculture include the need for improved yields, tolerance to biotic and abiotic stresses, and improved nutrition. The next generation of biotechnology-derived crops may utilize regulatory proteins, such as transcription factors that modulate gene expression and/or endogenous plant pathways. In this review, we discuss the applicability of the current safety assessment paradigm to biotechnology-derived crops developed using modifications involving regulatory proteins. The growing literature describing the molecular biology underlying plant domestication and conventional breeding demonstrates the naturally occurring genetic variation found in plants, including significant variation in the classes, expression, and activity of regulatory proteins. Specific examples of plant modifications involving insertion or altered expression of regulatory proteins are discussed as illustrative case studies supporting the conclusion that the current comparative safety assessment process is appropriate for these types of biotechnology-developed crops. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 18602733 [PubMed - indexed for MEDLINE] 132: Plant Foods Hum Nutr. 2008 Sep;63(3):119-25. Epub 2008 Jul 4. Evaluation of the lime-cooking and tortilla making properties of quality protein maize hybrids grown in Mexico. Serna-Saldivar SO, Amaya Guerra CA, Herrera Macias P, Melesio Cuellar JL, Preciado Ortiz RE, Terron Ibarra AD, Vazquez Carrillo G. Departamento de Biotecnología e Ingeniería de Alimentos, Tecnológico de Monterrey, Av. Eugenio Garza Sada 2501 Sur, CP 64849, Monterrey, México. sserna@itesm.mx Eleven experimental and three commercial white quality protein maize (QPM) hybrids and two regular endosperm controls were planted at Celaya, Guanajuato, Mexico with the aim of comparing grain physical characteristics, protein quality, lime-cooking and tortilla making properties. All genotypes were planted under irrigation using a density of 80,000 plants/ha and fertilized with 250 kg N-60 P-60 K per hectare. When compared with the controls these QPM genotypes had lower test (77.4 vs. 76.5 kg/hL) and 1,000 kernel weights (327 vs. 307 g), softer endosperm texture (2.5 vs. 1.8 where 1 = soft, 2 intermediate and 3 hard endosperm), lower protein (10.0 vs. 8.0%), higher nixtamal water uptake after 30 min lime-cooking (50.0 vs. 53.1% moisture) and lower pericarp removal scores. The lower thousand-kernel weight and softer endosperm texture observed in the QPM genotypes lowered the optimum lime-cooking time as estimated with regression equations. Most QPM genotypes had higher amounts of lysine, tryptophan and albumins/globulins when compared with the controls. QPMs HEC 424973, HEC 774986 and HEC 734286 had the best grain traits for nixtamalization and therefore the best potential for industrial utilization. The commercial use of these QPM hybrids should benefit Mexicans who depend on tortillas as the main staple. Publication Types: Research Support, Non-U.S. Gov't PMID: 18600459 [PubMed - indexed for MEDLINE] 133: J Agric Food Chem. 2008 Aug 13;56(15):6648-55. Epub 2008 Jul 4. Comparison of sugar, acids, and volatile composition in raspberry bushy dwarf virus-resistant transgenic raspberries and the wild type 'meeker' (rubus idaeus L.). Malowicki SM, Martin R, Qian MC. Department of Food Science and Technology, Oregon State University, and USDA-ARS, Horticultural Crops Research Laboratory, 3420 N.W. Orchard Avenue, Corvallis, Oregon 97331, USA. Raspberry bushy dwarf virus (RBDV) causes a significant reduction in yield and quality in raspberry and raspberry-blackberry hybrid. Genetic modifications were made to 'Meeker' red raspberries to impart RBDV resistance. The RBDV-resistant transgenic and wild type 'Meeker' plants were grown in Oregon and Washington, and the fruits were harvested in the 2004 and 2005 growing seasons. Year-to-year and site-to-site variations were observed for the degrees Brix and titratable acidity, with Oregon raspberries having slightly higher degrees Brix and lower titratable acidity than Washington raspberries. Twenty-nine volatile compounds were quantified using stir bar sorptive extraction (SBSE) paired with gas chromatography-mass spectrometry (GC-MS). There were very few differences in volatile concentrations between the transgenic varieties and the wild type 'Meeker'. Much larger variations were observed between sites and harvest seasons. Raspberries grown in Oregon appeared to have higher concentrations of delta-octalactone, delta-decalactone, geraniol, and linalool. Chiral analysis of alpha-ionone, alpha-pinene, linalool, terpinen-4-ol, delta-octalactone, and delta-decalactone demonstrated a much higher percentage of one isomer over the other, particularly alpha-ionone, alpha-pinene, delta-octalactone, and delta-decalactone, with more than 90% of one isomer, while a racemic mixture was observed for linalool. The isomeric analysis revealed very little variation between varieties, locations, or years. The flavor compounds tested in this study did not show any difference between the transgenic lines and the wild type 'Meeker' raspberry. Publication Types: Comparative Study PMID: 18598047 [PubMed - indexed for MEDLINE] 134: PLoS ONE. 2008 Jul 2;3(7):e2601. Maternal feeding controls fetal biological clock. Ohta H, Xu S, Moriya T, Iigo M, Watanabe T, Nakahata N, Chisaka H, Hanita T, Matsuda T, Ohura T, Kimura Y, Yaegashi N, Tsuchiya S, Tei H, Okamura K. Center for Perinatal Medicine, Tohoku University Hospital, Sendai, Japan. hideohta@mail.tains.tohoku.ac.jp BACKGROUND: It is widely accepted that circadian physiological rhythms of the fetus are affected by oscillators in the maternal brain that are coupled to the environmental light-dark (LD) cycle. METHODOLOGY/PRINCIPAL FINDINGS: To study the link between fetal and maternal biological clocks, we investigated the effects of cycles of maternal food availability on the rhythms of Per1 gene expression in the fetal suprachiasmatic nucleus (SCN) and liver using a transgenic rat model whose tissues express luciferase in vitro. Although the maternal SCN remained phase-locked to the LD cycle, maternal restricted feeding phase-advanced the fetal SCN and liver by 5 and 7 hours respectively within the 22-day pregnancy. CONCLUSIONS/SIGNIFICANCE: Our results demonstrate that maternal feeding entrains the fetal SCN and liver independently of both the maternal SCN and the LD cycle. This indicates that maternal-feeding signals can be more influential for the fetal SCN and particular organ oscillators than hormonal signals controlled by the maternal SCN, suggesting the importance of a regular maternal feeding schedule for appropriate fetal molecular clockwork during pregnancy. Publication Types: Research Support, Non-U.S. Gov't PMID: 18596966 [PubMed - indexed for MEDLINE] 135: Arch Latinoam Nutr. 2008 Mar;58(1):49-58. [Differentiated perception of transgenic tomato sauce in the southern Chile] [Article in Spanish] Schnettler Morales B, Sepúlveda Bravo O, Ruiz Fuentes D, Denegri Coria M. Departamento de Producción Agropecuaria, Facultad de Ciencias Agropecuarias y Forestales, Universidad de La Frontera, Chile. The present study considers the debate generated in developed countries by genetically modified foods, the importance of this variable to consumers in Temuco (Araucanía Region, Chile) when purchasing tomato sauce and different market segments were studied through a personal survey administered to 400 people. Using conjoint analysis, it was determined that the presence of genetic modification in food was generally more important than the brand and purchase price. Using cluster analysis, three segments were distinguished, with the most numerous (49.3%) placing the greatest importance on the presence of genetic modification (GM) in food and rejecting the transgenic product. The second group (39.4%) gave the greatest importance to the brand and preferred tomato sauce with genetically modified ingredients. The smallest segment (11.3%) placed the greatest value on price and preferred transgenic tomato sauce. The three segments prefer the national brand, reject the store brand and react positively to lower prices. The segment sensitive to the presence of GM in food comprised mainly those younger than 35 years of age, single and with no children. The absence of GM in food of vegetable origin is desirable for young consumers in the Araucanía Region, but a significant proportion accepts genetic modification in food (50.7%). Publication Types: English Abstract PMID: 18589572 [PubMed - indexed for MEDLINE] 136: Curr Biol. 2008 Jun 3;18(11):R446-7. Commodity prices push European policies. Williams N. Publication Types: News PMID: 18584807 [PubMed - indexed for MEDLINE] 137: Anal Chem. 2008 Aug 1;80(15):6127-30. Epub 2008 Jun 24. High-throughput polymerase chain reaction in parallel circular loops using magnetic actuation. Sun Y, Nguyen NT, Kwok YC. National Institute of Education, Nanyang Technological University, 01 Nanyang Walk, Singapore 637616. We report here a novel multichannel closed-loop magnetically actuated microchip for high-throughput polymerase chain reaction (PCR). This is achieved by designing a series of concentric circular channels on one microchip and exploiting a magnetic force to drive DNA samples flowing continuously through the closed loops. The magnetic force arises from an external permanent magnet through ferrofluid plugs inside the microchannels. The magnet enables simultaneous actuation of DNA samples in all the channels. As the samples go around the loops, they pass through three preset temperature zones. Parameters of PCR, such as incubation time, temperatures, and number of cycles, can be fully controlled and adjusted. High reproducibility was achieved for different channels in the same run and for the same channels in consecutive runs. Genetically modified organisms (GMOs) were amplified simultaneously using the developed device. This simple, reliable, and high-throughput PCR microchip would find wide applications in forensic, clinical, and biological fields. Publication Types: Research Support, Non-U.S. Gov't PMID: 18572956 [PubMed - indexed for MEDLINE] 138: J Agric Food Chem. 2008 Jul 23;56(14):5514-20. Epub 2008 Jun 21. Development of one novel multiple-target plasmid for duplex quantitative PCR analysis of roundup ready soybean. Zhang H, Yang L, Guo J, Li X, Jiang L, Zhang D. GMO Detection Laboratory, SJTU-Bor Luh Food Safety Center, Shanghai Jiao Tong University, Shanghai 200240, People's Republic of China To enforce the labeling regulations of genetically modified organisms (GMOs), the application of reference molecules as calibrators is becoming essential for practical quantification of GMOs. However, the reported reference molecules with tandem marker multiple targets have been proved not suitable for duplex PCR analysis. In this study, we developed one unique plasmid molecule based on one pMD-18T vector with three exogenous target DNA fragments of Roundup Ready soybean GTS 40-3-2 (RRS), that is, CaMV35S, NOS, and RRS event fragments, plus one fragment of soybean endogenous Lectin gene. This Lectin gene fragment was separated from the three exogenous target DNA fragments of RRS by inserting one 2.6 kb DNA fragment with no relatedness to RRS detection targets in this resultant plasmid. Then, we proved that this design allows the quantification of RRS using the three duplex real-time PCR assays targeting CaMV35S, NOS, and RRS events employing this reference molecule as the calibrator. In these duplex PCR assays, the limits of detection (LOD) and quantification (LOQ) were 10 and 50 copies, respectively. For the quantitative analysis of practical RRS samples, the results of accuracy and precision were similar to those of simplex PCR assays, for instance, the quantitative results were at the 1% level, the mean bias of the simplex and duplex PCR were 4.0% and 4.6%, respectively, and the statistic analysis ( t-test) showed that the quantitative data from duplex and simplex PCR had no significant discrepancy for each soybean sample. Obviously, duplex PCR analysis has the advantages of saving the costs of PCR reaction and reducing the experimental errors in simplex PCR testing. The strategy reported in the present study will be helpful for the development of new reference molecules suitable for duplex PCR quantitative assays of GMOs. Publication Types: Research Support, Non-U.S. Gov't PMID: 18570432 [PubMed - indexed for MEDLINE] 139: Crit Rev Food Sci Nutr. 2008 Jun;48(6):553-98. A review of rice authenticity/adulteration methods and results. Vlachos A, Arvanitoyannis IS. School of Agricultural Sciences, Department of Agriculture Icthyology and Aquatic Environment, Fytokou Street Nea Ionia Magnesias, University of Thessaly, Volos, Hellas, Greece. Rice importance resides in its high consumption mainly in Asia and Africa and less in the EU. Several cultivars, both GM and non-GM, have established themselves in various regions depending mainly on the climatic and soil conditions. A high number of analytical, enzymic, and genomic analyses (instrumental) in conjunction with sensory analysis were applied (not always very successfully) towards detecting deliberate or non-deliberate rice adulteration. It was shown that the application of multivariate analysis to data obtained is very beneficial because it allows the effective discrimination of different origin, and/or cultivar rice. Although sensory analysis is based on a trained panel (subjective method), if this panel has been properly trained the adulteration results are comparable to those of the instrumental analysis obtained. Publication Types: Review PMID: 18568860 [PubMed - indexed for MEDLINE] 140: Transgenic Res. 2008 Dec;17(6):1117-29. Epub 2008 Jun 18. Development and evaluation of transgenic rice seeds accumulating a type II-collagen tolerogenic peptide. Hashizume F, Hino S, Kakehashi M, Okajima T, Nadano D, Aoki N, Matsuda T. Department of Applied Molecular Biosciences, Graduate School of Bioagricultural Sciences, Nagoya University, Chikusa-ku, Nagoya, Aichi 464-8601, Japan. Type II collagen (CII) in joint cartilage is known to be a major auto-antigen in human rheumatoid arthritis. Several animal model- and clinical-studies on tolerance-based immunotherapy for the arthritis have been conducted by administrating synthetic immunodominant peptides through an oral route. In the present study, to produce a tolerogenic peptide with therapeutic potential in transgenic rice plants, a gene construct producing glutelin fusion protein with tandem four repeats of a CII(250-270) peptide (residues 250-270) (GluA-4XCII(250-270)) containing a human T-cell epitope was introduced with a selection marker, hygromycin phosphotransferase gene (hygromycin-resistance gene) (hph), by co-transformation. Several transgenic plants with high and stable expression of gluA-4XCII ( 250-270 ), but no hph, were selected based on both DNA and protein analyses. The GluA-4XCII(250-270) fusion proteins were detected as both precursor and processed forms mainly in a glutelin fraction of rice endosperm protein extracts and in protein-body rich fractions prepared by density gradient ultracentrifugation. The amount of accumulated CII(250-270) peptide was immunochemically estimated to be about 1 microg per seed. Feeding DBA/1 mice the transgenic rice seeds (25 microg of the peptide per mouse a day) for 2 weeks showed tendencies lowering and delaying serum specific-IgG2a response against subsequent and repeated intraperitoneal-injection of type II collagen. Taken these together, the CII-immunodominant peptide could effectively be produced and accumulated as a glutelin-fusion protein in the transgenic rice seeds, which might be useful as pharmaceutical materials and functional food for prevention and therapy for anti-CII autoimmune diseases like human rheumatoid arthritis. Publication Types: Research Support, Non-U.S. Gov't PMID: 18563612 [PubMed - indexed for MEDLINE] 141: Nature. 2008 Jun 19;453(7198):979. Europe needs to protect its transgenic crop research. Atkinson HJ, Urwin PE. Publication Types: Letter PMID: 18563128 [PubMed - indexed for MEDLINE] 142: Res Vet Sci. 2008 Jun;84(3):395-408. Atlantic salmon (Salmo salar L.) parr fed genetically modified soybeans and maize: Histological, digestive, metabolic, and immunological investigations. Bakke-McKellep AM, Sanden M, Danieli A, Acierno R, Hemre GI, Maffia M, Krogdahl A. Aquaculture Protein Centre (APC), CoE, Norway. anne.mckellep@veths.no Physiological and health related responses to dietary inclusion of genetically modified (GM) full-fat soybean meal (Roundup Ready; GM-soy) and maize (MON810 Bt-maize; GM-maize), as well as non-parental, untransformed lines (nGM-soy and nGM-maize D2), were evaluated in farmed Atlantic salmon (Salmo salar L.) parr during the first 8 months of feeding. Significant effects of dietary GM presence were only found in intestinal Na+-dependent d-glucose uptake and SGLT1 protein level in the region pyloric caeca in which the highest values were found in the GM-soy, intermediate in the nGM-soy, and lowest in the standard FM fed groups. Data from this study confirm that GM soybeans (RRS) and maize (MON810) at inclusion levels of about 6% appear to be as safe as commercially available nGM soy and maize in diets for Atlantic salmon parr. Results from studies with higher inclusion levels and with non-modified, isogenic or near-isogenic parental lines as control groups are pending. Publication Types: Research Support, Non-U.S. Gov't PMID: 18561390 [PubMed - indexed for MEDLINE] 143: Curr Opin Allergy Clin Immunol. 2008 Jun;8(3):270-5. Soy allergy in perspective. Ballmer-Weber BK, Vieths S. Allergy Unit, Department of Dermatology, University Hospital Zurich, Zurich, Switzerland. barbara.ballmer@usz.ch PURPOSE OF REVIEW: The purpose of this paper is to review and discuss studies on soy allergy. RECENT FINDINGS: In Central Europe soy is a clinically relevant birch pollen-related allergenic food. Crossreaction is mediated by a Bet v 1 homologous protein, Gly m 4. Additionally, birch pollen allergic patients might acquire through Bet v 1 sensitization allergies to mungbean or peanut, in which Vig r 1 and Ara h 8 are the main cross-reactive allergens. Threshold doses in soy allergic individuals range from 10 mg to 50 g of soy and are more than one order of magnitude higher than in peanut allergy. No evidence was found for increased allergenicity of genetically modified soybeans. SUMMARY: In Europe, both primary and pollen-related food allergy exist. The diagnosis of legume allergy in birch pollen-sensitized patients should not be excluded on a negative IgE testing to legume extracts. Bet v 1 related allergens are often underrepresented in extracts. Gly m 4 from soy and Ara h 8 from peanut are nowadays commercially available and are recommended in birch pollen allergic patients with suspicion of soy or peanut allergy, but negative extract-based diagnostic tests to screen for IgE specific to these recombinant allergens. Publication Types: Review PMID: 18560305 [PubMed - indexed for MEDLINE] 144: Plant Foods Hum Nutr. 2008 Sep;63(3):111-7. Epub 2008 Jun 13. Changes caused by genotype and environmental conditions in beta-glucan content of spring barley for dietetically beneficial human nutrition. Ehrenbergerová J, Brezinová Belcredi N, Psota V, Hrstková P, Cerkal R, Newman CW. Department of Crop Science, Breeding and Plant Medicine, Mendel University of Agriculture and Forestry in Brno, Zemedelska 1, 613 00, Brno, Czech Republic. ehren@mendelu.cz Over the 5-year period (2000-2004), a significantly higher beta-glucan content was detected in the waxy varieties Washonubet, Wabet, and Wanubet (6.8-7.6%) and lines formed by crossing these varieties with malting varieties (5.8-7.1%). Conversely, the non-waxy hulled malting-type varieties Kompakt (4.0%) and Krona (4.3%) had significantly lower contents of beta-glucan. The observations also showed that concentrations of beta-glucans in 2000-2004 were significantly affected not only by varieties, but also environmental conditions in the growing periods and interactions of these two factors. Higher precipitation during the flowering time and grain filling period and lower temperatures during the flowering time in 2002 had negative effects on concentration of beta-glucans. Conversely, drier and warmer weather in 2003 enhanced the content of beta-glucans. The results show that it is possible to increase the content of beta-glucan in spring barley grain by implementing selective breeding practices. Compared to the parental malting varieties, the mean content of beta-glucans in F(4)-F(8) generations was increased by 1.8 and 2.0% by recombination in lines Kompakt x Wabet and Wanubet x Krona, respectively. Significant effect of environmental conditions and their interactions with varieties indicated the necessity to assess standard qualities of barley as a food material. Publication Types: Research Support, Non-U.S. Gov't PMID: 18551369 [PubMed - indexed for MEDLINE] 145: BMC Plant Biol. 2008 Jun 12;8:65. The effects of enhanced methionine synthesis on amino acid and anthocyanin content of potato tubers. Dancs G, Kondrák M, Bánfalvi Z. Agricultural Biotechnology Center, P,O, Box 411, H-2101 Gödöllõ, Hungary. dancs@abc.hu BACKGROUND: Potato is a staple food in the diet of the world's population and also being used as animal feed. Compared to other crops, however, potato tubers are relatively poor in the essential amino acid, methionine. Our aim was to increase the methionine content of tubers by co-expressing a gene involved in methionine synthesis with a gene encoding a methionine-rich storage protein in potato plants. RESULTS: In higher plants, cystathionine gamma-synthase (CgS) is the first enzyme specific to methionine biosynthesis. We attempted to increase the methionine content of tubers by expressing the deleted form of the Arabidopsis CgS (CgSDelta90), which is not regulated by methionine, in potato plants. To increase the incorporation of free methionine into a storage protein the CgSDelta90 was co-transformed with the methionine-rich 15-kD beta-zein. Results demonstrated a 2- to 6-fold increase in the free methionine content and in the methionine content of the zein-containing protein fraction of the transgenic tubers. In addition, in line with higher methionine content, the amounts of soluble isoleucine and serine were also increased. However, all of the lines with high level of CgSDelta90 expression were phenotypically abnormal showing severe growth retardation, changes in leaf architecture and 40- to 60% reduction in tuber yield. Furthermore, the colour of the transgenic tubers was altered due to the reduced amounts of anthocyanin pigments. The mRNA levels of phenylalanine ammonia-lyase (PAL), the enzyme catalysing the first step of anthocyanin synthesis, were decreased. CONCLUSION: Ectopic expression of CgSDelta90 increases the methionine content of tubers, however, results in phenotypic aberrations in potato. Co-expression of the 15-kD beta-zein with CgSDelta90 results in elevation of protein-bound methionine content of tubers, but can not overcome the phenotypical changes caused by CgSDelta90 and can not significantly improve the nutritional value of tubers. The level of PAL mRNA and consequently the amount of anthocyanin pigments are reduced in the CgSDelta90 transgenic tubers suggesting that methionine synthesis and production of anthocyanins is linked. Publication Types: Research Support, Non-U.S. Gov't PMID: 18549488 [PubMed - indexed for MEDLINE] 146: Anal Bioanal Chem. 2008 Oct;392(3):355-67. Epub 2008 Jun 8. New trends in bioanalytical tools for the detection of genetically modified organisms: an update. Michelini E, Simoni P, Cevenini L, Mezzanotte L, Roda A. Department of Pharmaceutical Sciences, University of Bologna, via Belmeloro 6, 40126, Bologna, Italy. Despite the controversies surrounding genetically modified organisms (GMOs), the production of GM crops is increasing, especially in developing countries. Thanks to new technologies involving genetic engineering and unprecedented access to genomic resources, the next decade will certainly see exponential growth in GMO production. Indeed, EU regulations based on the precautionary principle require any food containing more than 0.9% GM content to be labeled as such. The implementation of these regulations necessitates sampling protocols, the availability of certified reference materials and analytical methodologies that allow the accurate determination of the content of GMOs. In order to qualify for the validation process, a method should fulfil some criteria, defined as "acceptance criteria" by the European Network of GMO Laboratories (ENGL). Several methods have recently been developed for GMO detection and quantitation, mostly based on polymerase chain reaction (PCR) technology. PCR (including its different formats, e.g., double competitive PCR and real-time PCR) remains the technique of choice, thanks to its ability to detect even small amounts of transgenes in raw materials and processed foods. Other approaches relying on DNA detection are based on quartz crystal microbalance piezoelectric biosensors, dry reagent dipstick-type sensors and surface plasmon resonance sensors. The application of visible/near-infrared (vis/NIR) spectroscopy or mass spectrometry combined with chemometrics techniques has also been envisaged as a powerful GMO detection tool. Furthermore, in order to cope with the multiplicity of GMOs released onto the market, the new challenge is the development of routine detection systems for the simultaneous detection of numerous GMOs, including unknown GMOs. Publication Types: Review PMID: 18537027 [PubMed - indexed for MEDLINE] 147: Ecol Appl. 2008 Jun;18(4):826-37. Brazilian free-tailed bats as insect pest regulators in transgenic and conventional cotton crops. Federico P, Hallam TG, McCracken GF, Purucker ST, Grant WE, Correa-Sandoval AN, Westbrook JK, Medellin RA, Cleveland CJ, Sansone CG, López JD Jr, Betke M, Moreno-Valdez A, Kunz TH. Department of Ecology and Evolutionary Biology, 569 Dabney Hall, University of Tennessee, Knoxville, Tennessee 37996, USA. During the past 12000 years agricultural systems have transitioned from natural habitats to conventional agricultural regions and recently to large areas of genetically engineered (GE) croplands. This GE revolution occurred for cotton in a span of slightly more than a decade during which a switch occurred in major cotton production areas from growing 100% conventional cotton to an environment in which 95% transgenics are grown. Ecological interactions between GE targeted insects and other insectivorous insects have been investigated. However, the relationships between ecological functions (such as herbivory and ecosystem transport) and agronomic benefits of avian or mammalian insectivores in the transgenic environment generally remain unclear, although the importance of some agricultural pest management services provided by insectivorous species such as the Brazilian free-tailed bat, Tadarida brasiliensis, have been recognized. We developed a dynamic model to predict regional-scale ecological functions in agricultural food webs by using the indicators of insect pest herbivory measured by cotton boll damage and insect emigration from cotton. In the south-central Texas Winter Garden agricultural region we find that the process of insectivory by bats has a considerable impact on both the ecology and valuation of harvest in Bacillus thuringiensis (Bt) transgenic and nontransgenic cotton crops. Predation on agricultural pests by insectivorous bats may enhance the economic value of agricultural systems by reducing the frequency of required spraying and delaying the ultimate need for new pesticides. In the Winter Garden region, the presence of large numbers of insectivorous bats yields a regional summer dispersion of adult pest insects from Bt cotton that is considerably reduced from the moth emigration when bats are absent in either transgenic or non-transgenic crops. This regional decrease of pest numbers impacts insect herbivory on a transcontinental scale. With a few exceptions, we find that the agronomics of both Bt and conventional cotton production is more profitable when large numbers of insectivorous bats are present. Publication Types: Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. PMID: 18536245 [PubMed - indexed for MEDLINE] 148: Nature. 2008 Jun 5;453(7196):707. The four-year fight for biological art. Steven Kurtz interviewed by Rachel Courtland. Kurtz S. Publication Types: Biography Historical Article Interview Personal Name as Subject: Kurtz S PMID: 18528358 [PubMed - indexed for MEDLINE] 149: Arch Latinoam Nutr. 2007 Dec;57(4):313-5. [Orthorexia or when a healthy diet becomes an obsession] [Article in Spanish] Bartrina JA. Departamento de Medicina Preventiva y Salud Pública, Universidad de Navarra, España. Orthorexia is an obsessive-compulsive process characterized by extreme care for and selection of what is considered to be pure 'healthy' food. This ritual leads to a very restrictive diet and social isolation as a compensation. Orthorexics obsessively avoid foods which may contain artificial colours, flavours, preservant agents, pesticide residues or genetically modified ingredients, unhealthy fats, foods containing too much salt or too much sugar and other components. The way of preparation, kitchenware and other tools used are also part of the obsessive ritual. People with orthorexia often have a history or features in common with anorexic patients. They are very careful, detailed and tidy persons with an exagerated need for selfcare and protection. Women, adolescents and those who practice sports suchs as bodybuidling or ahthetics are the gruops at higher risk. A short test has been suggested as a screening tool and useful for early diagnosis of the disorder. Treatment of orthorexia require a multidisciplinary team involving physicians, psychoterapists and dietitians. In some cases, antiserotoninergic drugs may be required as part of the treatment. Publication Types: English Abstract PMID: 18524314 [PubMed - indexed for MEDLINE] 150: BMC Mol Biol. 2008 Jun 4;9:54. A novel universal real-time PCR system using the attached universal duplex probes for quantitative analysis of nucleic acids. Yang L, Liang W, Jiang L, Li W, Cao W, Wilson ZA, Zhang D. GMO detection laboratory, SJTU-Bor Luh Food Safety Center, School of life Science and Biotechnology, Shanghai Jiao Tong University, 800 Dongchuan Road, Shanghai 200240, PR China. yylltt@sjtu.edu.cn BACKGROUND: Real-time PCR techniques are being widely used for nucleic acids analysis, but one limitation of current frequently employed real-time PCR is the high cost of the labeled probe for each target molecule. RESULTS: We describe a real-time PCR technique employing attached universal duplex probes (AUDP), which has the advantage of generating fluorescence by probe hydrolysis and strand displacement over current real-time PCR methods. AUDP involves one set of universal duplex probes in which the 5' end of the fluorescent probe (FP) and a complementary quenching probe (QP) lie in close proximity so that fluorescence can be quenched. The PCR primer pair with attached universal template (UT) and the FP are identical to the UT sequence. We have shown that the AUDP technique can be used for detecting multiple target DNA sequences in both simplex and duplex real-time PCR assays for gene expression analysis, genotype identification, and genetically modified organism (GMO) quantification with comparable sensitivity, reproducibility, and repeatability with other real-time PCR methods. CONCLUSION: The results from GMO quantification, gene expression analysis, genotype identification, and GMO quantification using AUDP real-time PCR assays indicate that the AUDP real-time PCR technique has been successfully applied in nucleic acids analysis, and the developed AUDP real-time PCR technique will offer an alternative way for nucleic acid analysis with high efficiency, reliability, and flexibility at low cost. Publication Types: Evaluation Studies Research Support, Non-U.S. Gov't PMID: 18522756 [PubMed - indexed for MEDLINE] 151: BMC Bioinformatics. 2008 Jun 4;9:260. GMDD: a database of GMO detection methods. Dong W, Yang L, Shen K, Kim B, Kleter GA, Marvin HJ, Guo R, Liang W, Zhang D. GMO Detection Laboratory, SJTU-Bor Luh Food Safety Center, Key Laboratory of Microbial Metabolism, Ministry of Education, School of Life Science and Biotechnology, Shanghai Jiao Tong University, Shanghai, PR China. dong_wei@sjtu.edu.cn BACKGROUND: Since more than one hundred events of genetically modified organisms (GMOs) have been developed and approved for commercialization in global area, the GMO analysis methods are essential for the enforcement of GMO labelling regulations. Protein and nucleic acid-based detection techniques have been developed and utilized for GMOs identification and quantification. However, the information for harmonization and standardization of GMO analysis methods at global level is needed. RESULTS: GMO Detection method Database (GMDD) has collected almost all the previous developed and reported GMOs detection methods, which have been grouped by different strategies (screen-, gene-, construct-, and event-specific), and also provide a user-friendly search service of the detection methods by GMO event name, exogenous gene, or protein information, etc. In this database, users can obtain the sequences of exogenous integration, which will facilitate PCR primers and probes design. Also the information on endogenous genes, certified reference materials, reference molecules, and the validation status of developed methods is included in this database. Furthermore, registered users can also submit new detection methods and sequences to this database, and the newly submitted information will be released soon after being checked. CONCLUSION: GMDD contains comprehensive information of GMO detection methods. The database will make the GMOs analysis much easier. Publication Types: Research Support, Non-U.S. Gov't PMID: 18522755 [PubMed - indexed for MEDLINE] 152: EMBO Rep. 2008 Jun;9(6):500-4. GM directive deficiencies in the European Union. The current framework for regulating GM crops in the EU weakens the precautionary principle as a policy tool. Morris SH, Spillane C. Genetics and Biotechnology Laboratory, Department of Biochemistry and Biosciences Institute, University College Cork, Ireland. shane.morris@student.ucc.ie PMID: 18516083 [PubMed - indexed for MEDLINE] 153: J R Soc Med. 2008 Jun;101(6):290-8. Comment in: J R Soc Med. 2008 Sep;101(9):435. Genetically modified plants and human health. Key S, Ma JK, Drake PM. Molecular Immunology Unit, Centre for Infection, Department of Cellular and Molecular Medicine, St George's University of London Cranmer Terrace, London SW17 0RE, UK. Genetically modified (or GM) plants have attracted a large amount of media attention in recent years and continue to do so. Despite this, the general public remains largely unaware of what a GM plant actually is or what advantages and disadvantages the technology has to offer, particularly with regard to the range of applications for which they can be used. From the first generation of GM crops, two main areas of concern have emerged, namely risk to the environment and risk to human health. As GM plants are gradually being introduced into the European Union there is likely to be increasing public concern regarding potential health issues. Although it is now commonplace for the press to adopt 'health campaigns', the information they publish is often unreliable and unrepresentative of the available scientific evidence. We consider it important that the medical profession should be aware of the state of the art, and, as they are often the first port of call for a concerned patient, be in a position to provide an informed opinion. This review will examine how GM plants may impact on human health both directly - through applications targeted at nutrition and enhancement of recombinant medicine production - but also indirectly, through potential effects on the environment. Finally, it will examine the most important opposition currently facing the worldwide adoption of this technology: public opinion. Publication Types: Review PMID: 18515776 [PubMed - indexed for MEDLINE] 154: Plant Physiol Biochem. 2008 Jul;46(7):647-54. Epub 2008 Apr 25. Manipulation of sinapine, choline and betaine accumulation in Arabidopsis seed: towards improving the nutritional value of the meal and enhancing the seedling performance under environmental stresses in oilseed crops. Huang J, Rozwadowski K, Bhinu VS, Schäfer U, Hannoufa A. Agriculture and Agri-Food Canada, Saskatoon Research Centre, 107 Science Place, Saskatoon, Saskatchewan SK S7N 0X2, Canada. Sinapoylcholine (sinapine) is the most abundant antinutritional phenolic compound in cruciferous seeds. The quaternary ammonium compounds, choline, betaine and N,N-dimethylglycine, reside along a biosynthetic pathway linked to the synthesis of membrane phospholipids and neurotransmitters with various biological functions. In chicken, choline intake is required for optimal egg-laying performance and a choline supplement in diet is positively correlated with weight gains. A key step in sinapine biosynthesis is catalyzed by sinapoylglucose: choline sinapoyltransferase (SCT; EC 2.3.1.91) to form an ester linkage with sinapoylglucose and choline. The objective of this work was to reduce the sinapine content and simultaneously enhance free choline levels in cruciferous seeds. We report here the characterization of an Arabidopsis T-DNA insertion mutant lacking SCT activity in the seed. The sct mutant seeds contain less than 1% of sinapine and a more than 2-fold increase in free choline compared with wild type. We further expressed a choline oxidase (COX; EC 1.1.3.17) gene from Arthrobacter pascens in the Arabidopsis sct mutant and wild-type background using a napin gene promoter to convert free choline into betaine, an effective stress-alleviating compound in plants. Betaine was not detected in WT or sct mutant seeds. The sct+COX seeds contain nearly 2-fold greater levels of betaine relative to WT+COX seeds, demonstrating a positive correlation between endogenous choline and betaine production. In contrast, stable comparable levels of free choline were detected between sct+COX and WT+COX plants suggesting choline homeostasis likely prevent high levels of betaine production in the seed of transgenic COX plants. Publication Types: Research Support, Non-U.S. Gov't PMID: 18515127 [PubMed - indexed for MEDLINE] 155: Asia Pac J Clin Nutr. 2008;17 Suppl 1:241-4. Constantly evolving safety assessment protocols for GM foods. Sesikeran B, Vasanthi S. National Institute of Nutrition, (Indian Council of Medical Research), Jamai Osmanai PO, Tarnaka, Hyderabad - 500 007, India. dirnin_hyd@yahoo.co.in he introduction of GM foods has led to the evolution of a food safety assessment paradigm that establishes safety of the GM food relative to its conventional counterpart. The GM foods currently approved and marketed in several countries have undergone extensive safety testing under a structured safety assessment framework evolved by international organizations like FAO, WHO, Codex and OECD. The major elements of safety assessment include molecular characterization of inserted genes and stability of the trait, toxicity and allergenicity potential of the expressed substances, compositional analysis, potential for gene transfer to gut microflora and unintentional effects of the genetic modification. As more number and type of food crops are being brought under the genetic modification regime, the adequacy of existing safety assessment protocols for establishing safety of these foods has been questioned. Such crops comprise GM crops with higher agronomic vigour, nutritional or health benefit/ by modification of plant metabolic pathways and those expressing bioactive substances and pharmaceuticals. The safety assessment challenges of these foods are the potential of the methods to detect unintentional effects with higher sensitivity and rigor. Development of databases on food compositions, toxicants and allergens is currently seen as an important aid to development of safety protocols. With the changing global trends in genetic modification technology future challenge would be to develop GM crops with minimum amount of inserted foreign DNA so as to reduce the burden of complex safety assessments while ensuring safety and utility of the technology. Publication Types: Review PMID: 18296346 [PubMed - indexed for MEDLINE] 156: Asia Pac J Clin Nutr. 2008;17 Suppl 1:237-40. An Asian perspective on GMO and biotechnology issues. Teng PP. Natural Sciences & Science Education, National Institute of Education, Nanyang Technological University, Singapore 637616. paul.teng@nie.edu.sg Of the 102 million hectares that made up the global area of biotech crops in 2006, less than 8% (7.6 million ha) were in Asia. Three biotech crops are currently planted in significant areas in four Asian countries with government regulatory approval; namely, cotton, corn (maize), and canola. However, the amount of GM crop material imported into the Asian region for processing into food and animal feed is very substantial, and almost every country imports GM food. The issues which concern Asian scientists, regulators, and the lay public resemble those of other regions - biosafety, food safety, ethics and social justice, competitiveness, and the "EU" trade question. Most Asian countries now have regulatory systems for approving the commercialization of GM crops, and for approving food safety of GM crops. In Asia, because of the varied cultures, issues concerning the use of genes derived from animals arouse much emotion for religious and diet choice reasons. Because many Asian producers and farmers are small-scale, there is also concern about technology dependency and to whom the benefits accrue. All consumers surveyed have expressed concern about potential allergenic and long-term toxic effects, neither of which is grounded on scientific facts. Because of Asia's growing demand for high volumes of quality food, it is likely that GM crops will become an increasing feature of our diet. Publication Types: Review PMID: 18296345 [PubMed - indexed for MEDLINE] 157: Asia Pac J Clin Nutr. 2008;17 Suppl 1:233-6. International development of methods of analysis for the presence of products of modern biotechnology. Cantrill RC. AOCS, 2710 S. Boulder Dr., Urbana, IL 61802, USA. Richard.Cantrill@aocs.org Methods of analysis for products of modern biotechnology are required for national and international trade in seeds, grain and food in order to meet the labeling or import/export requirements of different nations and trading blocks. Although many methods were developed by the originators of transgenic events, governments, universities, and testing laboratories, trade is less complicated if there exists a set of international consensus-derived analytical standards. In any analytical situation, multiple methods may exist for testing for the same analyte. These methods may be supported by regional preferences and regulatory requirements. However, tests need to be sensitive enough to determine low levels of these traits in commodity grain for regulatory purposes and also to indicate purity of seeds containing these traits. The International Organization for Standardization (ISO) and its European counterpart have worked to produce a suite of standards through open, balanced and consensus-driven processes. Presently, these standards are approaching the time for their first review. In fact, ISO 21572, the "protein standard" has already been circulated for systematic review. In order to expedite the review and revision of the nucleic acid standards an ISO Technical Specification (ISO/TS 21098) was drafted to set the criteria for the inclusion of precision data from collaborative studies into the annexes of these standards. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 18296344 [PubMed - indexed for MEDLINE] 158: Asia Pac J Clin Nutr. 2008;17 Suppl 1:229-32. Nutritional and safety assessment of foods and feeds nutritionally improved through biotechnology--case studies by the International Food Biotechnology Committee of ILSI. Glenn KC. Monsanto Co., O3B, 800 North Lindbergh Blvd., St. Louis, MO 63167 USA. Kevin.c.glenn@monsanto.com During the last two decades, the public and private sectors have made substantial research progress internationally toward improving the nutritional value of a wide range of food and feed crops. Nevertheless, significant numbers of people still suffer from the effects of undernutrition. As newly developed crops with nutritionally improved traits come closer to being available to producers and consumers, scientifically sound and efficient processes are needed to assess the safety and nutritional quality of these crops. In 2004, a Task Force of international scientific experts, convened by the International Food Biotechnology Committee (IFBiC) of ILSI, published recommendations for the safety and nutritional assessment of foods and feeds nutritionally improved through modern biotechnology (J. Food Science, 2004, 69:CRH62-CRH68). The comparative safety assessment process is a basic principle in this publication and is the starting point, not the conclusion, of the analysis. Significant differences in composition are expected to be observed in the case of nutritionally enhanced crops and must be assessed on a case-by-case basis. The Golden Rice 2 case study will be presented as an example of a food crop nutritionally enhanced through the application of modern biotechnology (i.e., recombinant DNA techniques) to illustrate how the 2004 recommendations provide a robust paradigm for the safety assessment of "real world" examples of improved nutrition crops. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 18296343 [PubMed - indexed for MEDLINE] 159: Asia Pac J Clin Nutr. 2008;17 Suppl 1:99-102. Supplementing iron bioavailability enhanced mung bean. Purushothaman V, M A, Tsou SC, S S. Faculty of Community Education and Entrepreneurship Development, Avinashilingam University for Women, Coimba-tore- 641 043, Tamil Nadu, India. drvijip@yahoo.com.in Iron deficiency anaemia is a major public health problem. The high incidence is either due to insufficient intake of iron or poor bio availability. Enhancing the bio availability is as important as increasing the intake. The absorption could be enhanced by including ascorbic acid and beta carotene containing fruits and vegetables into recipes of iron containing food preparations. The effect of supplementation of iron bio-availability enhanced mung bean preparations was studied on 75 women who were compared against 75 who served as controls and another 75 who consumed regular traditional recipes. The methodology included identification of suitable mung bean variety, assessing iron in vitro bio availability, mapping the anaemic women, estimating their iron levels, supplementation for one year and studying the effect of supplementation. Mung bean supplementation had increased serum protein levels from 5.36 to 6.73 g/dl, serum iron levels had increased from 16.6 to 46.7 microg/dl. The TIBC levels decreased from 555 to 508 microg/dl while serum ferritin levels increased from 3.56 to 5.94 microg/dl and Hb levels from 7.54 to 8.29 g/dl. Thus, improving the bioavailability of iron of food preparations, will improve the iron status of women. Publication Types: Research Support, Non-U.S. Gov't PMID: 18296312 [PubMed - indexed for MEDLINE] 160: Asia Pac J Clin Nutr. 2008;17 Suppl 1:95-8. Global perspective of health related edible plants from the agricultural point of view. Lee YY, Tsou CS, Lin HC, Ien CH, Wu YT. Science and Technology Policy Research and Information Center, National Applied Research Laboratories, Taipei 10636, Taiwan. yylee@mail.stpi.org.tw In knowledge-based economies, nutrition concepts evolve with advances in agriculture. As people around the world become more health conscious, national health becomes one of the main directives for agricultural policies, including that of functional foods and their global markets. This article evaluates the development of the functional food industry in Taiwan and other countries through analysis of R&D capacity and bibliometrics. It attempts to identify future trends in nutrition with technology foresight research. Taiwan has a wide variety of indigenous herbal plants, although its functional food related literature is not large compared with some other Asian countries. However, there are quality papers on the immunologic functions of edible plants Globally there is much interest in edible plants with antioxidant activity and those phyto-nutrients which might help reduce the burden of chronic illness as well as in the nutrigenomics that will lead to the design of foods with these properties. To make the most of available agricultural resources, countries like Taiwan should relate agricultural development to the nutritional status of their populations. This strategy will add significant value to global agriculture. Publication Types: Review PMID: 18296311 [PubMed - indexed for MEDLINE] 161: Asia Pac J Clin Nutr. 2008;17 Suppl 1:91-4. Human health problems associated with current agricultural food production. Bhat RV. Secretary General, Federation of Asian Nutrition Societies, Centre for Science, Society and Culture M 11, Kakateeyanagar, Habshiguda, Hyderabad- 500 007, India. rameshvbhat@yahoo.com Scientific and technological developments in the agricultural sectors in the recent past has resulted in increased food production and at the same time led to certain public health concerns. Unseasonal rains at the time of harvest and improper post harvest technology often results in agricultural commodities being contaminated with certain fungi and results in the production of mycotoxins. Consumption of such commodities has resulted in human disease outbreaks. Naturally occurring toxins, inherently present in foods and either consumed as such or mixed up with grains, had been responsible for disease outbreaks. Other possible causes of health concern include the application of various agrochemicals such as pesticides and the use of antibiotics in aquaculture and veterinary practices. Foodborne pathogens entering the food chain during both traditional and organic agriculture pose a challenge to public health. Modern biotechnology, producing genetically modified foods, if not regulated appropriately could pose dangers to human health. Use of various integrated food management systems like the Hazard Analysis and critical control system approach for risk prevention, monitoring and control of food hazards are being emphasized with globalization to minimise the danger posed to human health from improper agricultural practices. Publication Types: Review PMID: 18296310 [PubMed - indexed for MEDLINE] 162: Asia Pac J Clin Nutr. 2008;17 Suppl 1:87-90. Application of agricultural biotechnology to improve food nutrition and healthcare products. Sun SS. The Chinese University of Hong Kong, Department of Biology, G88 Science Centre South Block, Shatin, NT, Hong Kong, China. ssun@cuhk.edu.hk Crop plants provide essential food nutrients to humans and livestock, including carbohydrates, lipids, proteins, minerals and vitamins, directly or indirectly. The level and composition of food nutrients vary significantly in different food crops. As a result, plant foods are often deficient in certain nutrient components. Relying on a single food crop as source of nutrients thus will not achieve a balanced diet and results in malnutrition and deficiency diseases, especially in the developing countries, due mainly to poverty. The development and application of biotechnology offers opportunities and novel possibilities to enhance the nutritional quality of crops, particularly when the necessary genetic variability is not available. While initial emphasis of agricultural biotechnology has been placed on input traits of crops such as herbicide tolerance, insect resistance and virus resistance, increasing effort and promising proof-of-concept products have been made in output traits including enhancing the nutritional quality of crops since 1990s. Advancements in plant transformation and transgene expression also allow the use of plants as bioreactors to produce a variety of bio-products at large scale and low cost. Many proof-of-concept plant-derived healthcare products have been generated and several commercialized. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 18296309 [PubMed - indexed for MEDLINE] 163: Asia Pac J Clin Nutr. 2008;17 Suppl 1:24-9. National food fortification: a dialogue with reference to Asia: policy in evolution. Wahlqvist ML. Center for Health Policy Research and Development, National Health Research Institutes, 35 Keyan Road, Zhunan, Miaoli County, Taiwan 350, RCO. profmlw@nhri.org.tw Food fortification generally refers to the addition of micronutrients and other favourably bio-active food components to food-stuffs where there are recognised deficiencies in the target population. Each forticant has had or could have regulatory implications. It is understandable, although arguable, in the face of a limited food supply skewed, for the majority, in the direction of starchy staples of low essential nutrient density. Efforts, with plant breeding, to biofortify such foods are underway and likely to be safer, more sustainable and affordable than chemical additions. Unfortunately, with an increasingly refined and naturally tasteless food supply (salty, fatty, sugary and starchy), and where energy requirements are falling because of physical inactivity, micronutrient fortification is being used as a nutritional "fix-it' strategy. In Asia, there are several critical micro- nutrients. No one national fortification program can deal with all deficiencies is likely to be highly selective for the nutrients which have the greatest advocacy or are most recognisable. They also leave the other health promoting food properties like intactness, nutrient spectrum, and phytonutrient content un-addressed. A variety of food-stuffs, with different biological origins, is the preferred approach. Where an optimal food system is not in place, there may be justification for fortification if there is regular monitoring and surveillance of the food supply and health outcomes occurs; is a clear cost-risk-benefit advantage in such a strategy; are programs in place to improve the nutritional value of the basic food supply and is an "exit strategy' for the fortification program. Publication Types: Review PMID: 18296294 [PubMed - indexed for MEDLINE] 164: Shokuhin Eiseigaku Zasshi. 2008 Apr;49(2):63-9. DNA extraction method using a silica-base resin type kit for the detection of genetically modified papaya. Ohmori K, Tsuchiya H, Watanabe T, Akiyama H, Maitani T, Yamada T, Hirayama K, Satoh S. Chemistry Division, Kanagawa Prefectural Institute of Public Health, Chigasaki, Kanagawa, Japan. Genetically modified (GM) papaya has not yet been approved for importation into, or cultivation in the European Union (EU) and Japan. A DNA extraction method using the Qiagen DNeasy Plant Mini Kit (PM method) and a method using a buffer containing cetyltrimethyl ammonium bromide (CTAB method) have been adopted as the official Japanese methods for detecting GM foods. However, the amounts of DNA extracted from papaya by these methods are very low. Therefore, we investigated an extraction method to obtain a high yield of DNA from raw or freeze-dried fresh papaya using the Promega Wizard DNA Clean-Up Resin System (WCR). The incubation for the extraction was carried out at 58 degrees C without proteinase K for 15 min. The extract was applied to a mini-column, then the column was washed with 80% isopropyl alcohol, and genomic DNA adsorbed on the column was eluted with TE buffer. The WCR method gave a higher yield of genomic DNA, and was simpler and faster than the PM method or CTAB method. In addition, it could be used to extract genomic DNA from fresh papaya at various stages of ripeness. Based on these results, we propose that the present method using WCR is the most practical and useful way to extract genomic DNA for the purpose of detecting GM papaya. Publication Types: Research Support, Non-U.S. Gov't PMID: 18503240 [PubMed - indexed for MEDLINE] 165: Clin Exp Allergy. 2008 Jul;38(7):1095-99. What is a food allergen? Lucas JS, Atkinson RG. Division of Infection Inflammation and Repair, University of Southampton, Southampton, UK. jlucas1@soton.ac.uk With the increasing prevalence of allergies, accurate identification of allergens is a major priority for allergists, scientists, the food industry, and food regulators. Knowledge of allergens is essential for risk assessment of novel genetically modified (GM) foods, and to develop recombinant proteins for the treatment and diagnosis of allergies. This Opinion Paper considers the lack of standardization for the clinical and scientific assessment of proteins before they are labelled as allergens. Food allergens are being reported and recorded in allergen databases, with minimal or in some cases apparently no published justification. IgE binding, rather than clinically relevant reactivity, is inappropriately used to confirm allergenicity. Using kiwifruit as an example, the lack of rigor in identifying allergenic proteins is considered. PMID: 18498418 [PubMed - indexed for MEDLINE] 166: Adv Biochem Eng Biotechnol. 2008;111:229-64. Food and agricultural biotechnology: a summary and analysis of ethical concerns. Thompson PB, Hannah W. Department of Philosophy, Michigan State University, 48824-1320, East Lansing, MI 48824-1320, USA, thomp649@msu.edu The range of social and ethical concerns that have been raised in connection with food and agricultural biotechnology is exceedingly broad. Many of these deal with risks and possible outcomes that are not unique to crops or animals developed using recombinant DNA. Food safety, animal welfare, socio-economic and environmental impacts, as well as shifts in power relations or access to technology raise concerns that might be generalized to many technologies. These aspects of the controversy over biotechnology are analyzed below as elements of general technological ethics, and key norms or values pertinent to each of these categories are specified in some detail. However, a number of special concerns unique to the use of rDNA in manipulating plant and animal genomes have been raised, and these are reviewed as well. The chapter concludes by reviewing two broad policy strategies for responding to the issues, one involving labels and consumer consent, the other applying the precautionary principle. Publication Types: Review PMID: 18496654 [PubMed - indexed for MEDLINE] 167: Food Chem Toxicol. 2008 Jul;46(7):2517-24. Epub 2008 Apr 13. Results of a 13-week safety assurance study with rats fed grain from corn rootworm-protected, glyphosate-tolerant MON 88017 corn. Healy C, Hammond B, Kirkpatrick J. Monsanto Company, 800 North Lindbergh Blvd., St. Louis, MO 63167, United States. charles.e.healy@monsanto.com Presented are the results of a 13-week rat feeding study with grain from MON 88017 corn (brand name YieldGard VT Rootworm/RR2), protected from feeding damage caused by corn rootworm and tolerant to glyphosate, the active ingredient in Roundup agricultural herbicides. Corn rootworm protection is accomplished through the introduction of cryBb1 coding sequence from Bacillus thuringiensis into the corn genome for in planta production of a bioactive form of Cry3Bb1 protein. Also included in the genome is the coding sequence for the CP4 EPSPS protein from Agrobacterium sp. strain CP4 that confers glyphosate herbicidal tolerance. MON 88017 was formulated into rodent diets at 11 or 33% (w/w) levels with its near isogenic control at a level of 33% (w/w). Additionally, six diets containing grain from different conventional (non-biotechnology-derived), reference hybrids were formulated, each at 33% (w/w) levels of one of six reference grains. All diets were nutritionally balanced and conformed to PMI specifications for Certified LabDiet 5002 (PMI Certified LabDiet 5002 is a registered trademark of Purina Mills, Inc.). The responses of rats fed diets containing MON 88017 were comparable to those of rats fed a diet containing grain from its near isogenic control. This study complements extensive agronomic, compositional, and farm animal feeding studies with MON 88017 grain, confirming that it is as safe and nutritious as grain from existing commercial corn hybrids. PMID: 18492601 [PubMed - indexed for MEDLINE] 168: Anal Bioanal Chem. 2008 Oct;392(3):341-6. Epub 2008 May 17. Should genetically modified foods be abandoned on the basis of allergenicity? Bachas-Daunert S, Deo SK. Princeton University, Princeton, NJ 08544, USA. PMID: 18488210 [PubMed - indexed for MEDLINE] 169: Nat Rev Genet. 2008 Jun;9(6):458-63. Opposition to transgenic technologies: ideology, interests and collective action frames. Herring RJ. Department of Government, Cornell University, White Hall 313, Ithaca, New York 14853, USA. rjh5@cornell.edu Genetic engineering has enabled significant, accepted innovations in medicine and other fields. In agriculture, however, a global cognitive divide around 'genetically modified organisms' (GMOs) has limited the diffusion and scope of this technology. The framing of agricultural products of recombinant DNA technology as GMOs lacks biological coherence, but has proved to be a powerful frame for opposition. Disaggregating the concept of the 'GMO' is a necessary condition for confronting misconceptions that constrain the use of biotechnology in addressing imperatives of development and escalating challenges from nature, especially in less-industrialized nations. PMID: 18487989 [PubMed - indexed for MEDLINE] 170: Georgian Med News. 2008 Apr;(157):39-44. IgE-mediated food hypersensitivity disorders. Gotua M, Lomidze N, Dolidze N, Gotua T. Food allergy has become a serious health concern especially in developed countries in the past two decades. In general population approximately 4-6% of children and 1-3% of adults experience food allergy. The article reviews IgE-mediated food hypersensitivity disorders. Epidemiology, Mechanism, Clinical manifestations, Genetically modified crops (GMOs), Diagnosis, Prevention and Treatment of IgE-mediated food allergies are discussed. The investigations show that over 90% of IgE-mediated food allergies in childhood are caused by: cow's milk, hen's egg, soy, peanuts, tree nuts, wheat, fish and shellfish. Also the causes of food allergy are food additives, genetically modified crops. Risk factors for food-dependent exercise-induced anaphylaxis include asthma and previous allergic reactions to the causative food. Food allergy is one of the most common causes of systematic anaphylaxis and anaphylactoid reactions, with an annual incidence of four cases per million populations and estimated 500 deaths annually. In addition to gastrointestinal symptoms, individuals may experience urticaria, angioedema, atopic dermatitis, oral syndrome, asthma, rhinitis, conjunctivitis, hypotension, shock and cardiac arrhythmias, caused by the massive release of mediators from mast cells and basophiles. Diagnosis of food allergy is based on history, detailed dietary analysis, skin testing, measuring specific IgE in blood serum and challenge tests. Treatment and prevention includes: avoidance diet, application of auto-injectable epinephrine, H1 and H2 antihistamines, corticosteroids, antileukotrienes, prostaglandin synthetase inhibitors, cromolyn sodium, etc. Publication Types: Review PMID: 18487689 [PubMed - indexed for MEDLINE] 171: Appl Environ Microbiol. 2008 Jul;74(14):4381-9. Epub 2008 May 16. Effectiveness of Bacillus thuringiensis-transgenic chickpeas and the entomopathogenic fungus Metarhizium anisopliae in controlling Helicoverpa armigera (Lepidoptera: Noctuidae). Lawo NC, Mahon RJ, Milner RJ, Sarmah BK, Higgins TJ, Romeis J. Agroscope Reckenholz-Tänikon Research Station ART, Reckenholzstr. 191, 8046 Zurich, Switzerland. The use of genetically modified (Bt) crops expressing lepidopteran-specific Cry proteins derived from the soil bacterium Bacillus thuringiensis is an effective method to control the polyphagous pest Helicoverpa armigera. As H. armigera potentially develops resistance to Cry proteins, Bt crops should be regarded as one tool in integrated pest management. Therefore, they should be compatible with biological control. Bioassays were conducted to understand the interactions between a Cry2Aa-expressing chickpea line, either a susceptible or a Cry2A-resistant H. armigera strain, and the entomopathogenic fungus Metarhizium anisopliae. In a first concentration-response assay, Cry2A-resistant larvae were more tolerant of M. anisopliae than susceptible larvae, while in a second bioassay, the fungus caused similar mortalities in the two strains fed control chickpea leaves. Thus, resistance to Cry2A did not cause any fitness costs that became visible as increased susceptibility to the fungus. On Bt chickpea leaves, susceptible H. armigera larvae were more sensitive to M. anisopliae than on control leaves. It appeared that sublethal damage induced by the B. thuringiensis toxin enhanced the effectiveness of M. anisopliae. For Cry2A-resistant larvae, the mortalities caused by the fungus were similar when they were fed either food source. To examine which strain would be more likely to be exposed to the fungus, their movements on control and Bt chickpea plants were compared. Movement did not appear to differ among larvae on Bt or conventional chickpeas, as indicated by the number of leaflets damaged per leaf. The findings suggest that Bt chickpeas and M. anisopliae are compatible to control H. armigera. Publication Types: Research Support, Non-U.S. Gov't PMID: 18487396 [PubMed - indexed for MEDLINE] 172: J Sep Sci. 2008 Jun;31(10):1810-8. Time of flight versus ion trap MS coupled to CE to analyse intact proteins. Erny GL, León C, Marina ML, Cifuentes A. Institute of Industrial Fermentations (CSIC), Madrid, Spain. In this work, two different CE-MS instruments, namely, CE-ESI-IT-MS and CE-ESI-TOF-MS, applied to analyse intact proteins from complex samples are investigated. The aim of this work was to compare both instruments in terms of LOD, number of proteins detected, and precision and repeatability in the determination of the protein relative molecular mass. Results show that although CE-ESI-IT-MS provides cleaner MS spectra of intact proteins, CE-ESI-TOF-MS allows the identification of a higher number of proteins from complex matrices in an easier way. Performance in terms of peak area reproducibility, LOD and precision in the determination of the molecular mass were similar for both instruments. The usefulness of the optimised CE-ESI-IT-MS and CE-ESI-TOF-MS conditions was demonstrated by studying the zein-proteins composition of three natural maize lines and their corresponding transgenic lines, showing no significant differences. Publication Types: Research Support, Non-U.S. Gov't PMID: 18481325 [PubMed - indexed for MEDLINE] 173: Nature. 2008 May 15;453(7193):263. German universities bow to public pressure over GM crops. Schiermeier Q. Publication Types: News PMID: 18480774 [PubMed - indexed for MEDLINE] 174: Biomed Environ Sci. 2008 Feb;21(1):53-62. Detection of genetically modified crops by combination of multiplex PCR and low-density DNA microarray. Zhou PP, Zhang JZ, You YH, Wu YN. National Institute for Nutrition and Food Safety, Chinese Center for Disease Control and Prevention, Beijing 100050, China. OBJECTIVE: To develop a technique for simultaneous detection of various target genes in Roundup Ready soybean by combining multiplex PCR and low-density DNA microarray. METHODS: Two sets of the multiplex PCR system were used to amplify the target genes in genetically modified (GM) soybean. Seventeen capture probes (PCR products) and 17 pairs of corresponding primers were designed according to the genetic characteristics of Rroundup Ready soybean (GTS40-3-2), maize (Mon810, Nk603, GA21), canola (T45, MS1/RF1), and rice (SCK) in many identified GM crops. All of the probes were categorized and identified as species-specific probes. One negative probe and one positive control probe were used to assess the efficiency of all reactions, and therefore eliminate any false positive and negative results. After multiplex PCR reaction, amplicons were adulterated with Cy5-dUTP and hybridized with DNA microarray. The array was then scanned to display the specific hybridization signals of target genes. The assay was applied to the analysis of sample of certified transgenic soybean (Roundup Ready GTS40-3-2) and canola (MS1/RF1). RESULTS: A combination technique of multiplex PCR and DNA microarray was successfully developed to identify multi-target genes in Roundup Ready soybean and MS1/RF1 canola with a great specificity and reliability. Reliable identification of genetic characteristics of Roundup Ready of GM soybean from genetically modified crops was achieved at 0.5% transgenic events, indicating a high sensitivity. CONCLUSION: A combination technique of multiplex PCR and low-density DNA microarray can reliably detect and identify the genetically modified crops. Publication Types: Research Support, Non-U.S. Gov't PMID: 18478979 [PubMed - indexed for MEDLINE] 175: Nat Neurosci. 2008 Jun;11(6):676-82. Epub 2008 May 11. Drosophila TRPA channel modulates sugar-stimulated neural excitation, avoidance and social response. Xu J, Sornborger AT, Lee JK, Shen P. Department of Cellular Biology, University of Georgia, 500 D. W. Brooks Drive, Athens, Georgia 30602, USA. Drosophila melanogaster postfeeding larvae show food-averse migration toward food-free habitats before metamorphosis. This developmental switching from food attraction to aversion is regulated by a neuropeptide Y (NPY)-related brain signaling peptide. We used the fly larva model to delineate the neurobiological basis of age-restricted response to environmental stimuli. Here we provide evidence for a fructose-responsive chemosensory pathway that modulates food-averse migratory and social behaviors. We found that fructose potently elicited larval food-averse behaviors, and painless (pain), a transient receptor potential channel that is responsive to noxious stimuli, was required for the fructose response. A subset of pain-expressing sensory neurons have been identified that show pain-dependent excitation by fructose. Although evolutionarily conserved avoidance mechanisms are widely appreciated for their roles in stress coping and survival, their biological importance in animal physiology and development remains unknown. Our findings demonstrate how an avoidance mechanism is recruited to facilitate animal development. Publication Types: Research Support, N.I.H., Extramural PMID: 18469811 [PubMed - indexed for MEDLINE] 176: J Exp Bot. 2008;59(9):2337-46. Epub 2008 May 9. A field-grown transgenic tomato line expressing higher levels of polyamines reveals legume cover crop mulch-specific perturbations in fruit phenotype at the levels of metabolite profiles, gene expression, and agronomic characteristics. Neelam A, Cassol T, Mehta RA, Abdul-Baki AA, Sobolev AP, Goyal RK, Abbott J, Segre AL, Handa AK, Mattoo AK. USDA-ARS, Henry A. Wallace Beltsville Agricultural Research Center, Building 001, Beltsville, MD 20705-2350, USA. Genetic modification of crop plants to introduce desirable traits such as nutritional enhancement, disease and pest resistance, and enhanced crop productivity is increasingly seen as a promising technology for sustainable agriculture and boosting food production in the world. Independently, cultural practices that utilize alternative agriculture strategies including organic cultivation subscribe to sustainable agriculture by limiting chemical usage and reduced tillage. How the two together affect fruit metabolism or plant growth in the field or whether they are compatible has not yet been tested. Fruit-specific yeast S-adenosylmethionine decarboxylase (ySAMdc) line 579HO, and a control line 556AZ were grown in leguminous hairy vetch (Vicia villosa Roth) (HV) mulch and conventional black polyethylene (BP) mulch, and their fruit analysed. Significant genotypexmulch-dependent interactions on fruit phenotype were exemplified by differential profiles of 20 fruit metabolites such as amino acids, sugars, and organic acids. Expression patterns of the ySAMdc transgene, and tomato SAMdc, E8, PEPC, and ICDHc genes were compared between the two lines as a function of growth on either BP or HV mulch. HV mulch significantly stimulated the accumulation of asparagine, glutamate, glutamine, choline, and citrate concomitant with a decrease in glucose in the 556AZ fruits during ripening as compared to BP. It enables a metabolic system in tomato somewhat akin to the one in higher polyamine-accumulating transgenic fruit that have higher phytonutrient content. Finally, synergism was found between HV mulch and transgenic tomato in up-regulating N:C indicator genes PEPC and ICDHc in the fruit. Publication Types: Comparative Study Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. PMID: 18469323 [PubMed - indexed for MEDLINE] 177: Nat Biotechnol. 2008 May;26(5):500-1. Comment on: Nat Biotechnol. 2008 Mar;26(3):247. Off the rails or on the mark? Chataway J, Tait J, Wield D. Publication Types: Comment Letter PMID: 18464777 [PubMed - indexed for MEDLINE] 178: Nat Biotechnol. 2008 May;26(5):499-500. Comment on: Nat Biotechnol. 2008 Mar;26(3):247. Off the rails or on the mark? Heinemann JA. Publication Types: Comment Letter PMID: 18464776 [PubMed - indexed for MEDLINE] 179: Nat Biotechnol. 2008 May;26(5):482. GM grass trials blocked. Fox JL. Publication Types: News PMID: 18464761 [PubMed - indexed for MEDLINE] 180: Nat Biotechnol. 2008 May;26(5):478. EU to monitor for Chinese GM rice. Huggett B. Publication Types: News PMID: 18464757 [PubMed - indexed for MEDLINE] 181: Trends Biotechnol. 2008 Jul;26(7):353-8. Epub 2008 Apr 29. Regulating coexistence of GM and non-GM crops without jeopardizing economic incentives. Demont M, Devos Y. Africa Rice Center (WARDA), B.P. 96, Saint-Louis, Senegal. m.demont@cgiar.org The ongoing debate about the coexistence of genetically modified (GM) and non-GM crops in the European Union (EU) mainly focuses on preventive measures needed to keep the adventitious presence of GM material in non-GM products below established tolerance thresholds, as well as on issues covering questions of liability and the duty to redress the incurred economic harm once adventitious mixing in non-GM products has occurred. By contrast, the interplay between the economic incentives and costs of coexistence has attracted little attention. The current overemphasis on the technical aspects and cost of coexistence over its economic incentives might lead EU policy-makers to adopt too stringent and rigid regulations on coexistence. Therefore, we argue for flexible coexistence regulations that explicitly take into account the economic incentives for coexistence. Our arguments provide a timely and important framework for EU policy-makers, who are currently struggling to implement coherent coexistence regulations in all member states. PMID: 18453019 [PubMed - indexed for MEDLINE] 182: J Agric Food Chem. 2008 May 28;56(10):3438-43. Epub 2008 Apr 29. International collaborative study of the endogenous reference gene LAT52 used for qualitative and quantitative analyses of genetically modified tomato. Yang L, Zhang H, Guo J, Pan L, Zhang D. GMO Detection Laboratory, SJTU-Bor Luh Food Safety Center, Key Laboratory of Microbial Metabolism, Ministry of Education, School of Life Science and Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, People's Republic of China One tomato ( Lycopersicon esculentum) gene, LAT52, has been proved to be a suitable endogenous reference gene for genetically modified (GM) tomato detection in a previous study. Herein are reported the results of a collaborative ring trial for international validation of the LAT52 gene as endogenous reference gene and its analytical systems; 14 GMO detection laboratories from 8 countries were invited, and results were finally received from 13. These data confirmed the species specificity by testing 10 plant genomic DNAs, less allelic variation and stable single copy number of the LAT52 gene, among 12 different tomato cultivars. Furthermore, the limit of detection of LAT52 qualitative PCR was proved to be 0.1%, which corresponded to 11 copies of haploid tomato genomic DNA, and the limit of quantification for the quantitative PCR system was about 10 copies of haploid tomato genomic DNA with acceptable PCR efficiency and linearity. Additionally, the bias between the test and true values of 8 blind samples ranged from 1.94 to 10.64%. All of these validated results indicated that the LAT52 gene is suitable for use as an endogenous reference gene for the identification and quantification of GM tomato and its derivates. Publication Types: Research Support, Non-U.S. Gov't PMID: 18442244 [PubMed - indexed for MEDLINE] 183: J Neurosci Res. 2008 Aug 15;86(11):2553-63. Antipsychotic drugs up-regulate tryptophan hydroxylase in ADF neurons of Caenorhabditis elegans: role of calcium-calmodulin-dependent protein kinase II and transient receptor potential vanilloid channel. Donohoe DR, Phan T, Weeks K, Aamodt EJ, Dwyer DS. Department of Pharmacology, Toxicology and Neuroscience, Louisiana State University Health Sciences Center-Shreveport, Shreveport, Louisiana 71130, USA. Antipsychotic drugs produce acute behavioral effects through antagonism of dopamine and serotonin receptors, and long-term adaptive responses that are not well understood. The goal of the study presented here was to use Caenorhabditis elegans to investigate the molecular mechanism or mechanisms that contribute to adaptive responses produced by antipsychotic drugs. First-generation antipsychotics, trifluoperazine and fluphenazine, and second-generation drugs, clozapine and olanzapine, increased the expression of tryptophan hydroxylase-1::green fluorescent protein (TPH-1::GFP) and serotonin in the ADF neurons of C. elegans. This response was absent or diminished in mutant strains lacking the transient receptor potential vanilloid channel (TRPV; osm-9) or calcium/calmodulin-dependent protein kinase II (CaMKII; unc-43). The role of calcium signaling was further implicated by the finding that a selective antagonist of calmodulin and a calcineurin inhibitor also enhanced TPH-1::GFP expression. The ADF neurons modulate foraging behavior (turns/reversals off food) through serotonin production. We found that short-term exposure to the antipsychotic drugs altered the frequency of turns/reversals off food. This response was mediated through dopamine and serotonin receptors and was abolished in serotonin-deficient mutants (tph-1) and strains lacking the SER-1 and MOD-1 serotonin receptors. Consistent with the increase in serotonin in the ADF neurons induced by the drugs, drug withdrawal after 24-hr treatment was accompanied by a rebound in the number of turns/reversals, which demonstrates behavioral adaptation in serotonergic systems. Characterization of the cellular, molecular, and behavioral adaptations to continuous exposure to antipsychotic drugs may provide insight into the long-term clinical effects of these medications. Publication Types: Research Support, N.I.H., Extramural PMID: 18438926 [PubMed - indexed for MEDLINE] 184: Science. 2008 Apr 25;320(5875):473-5. Is the drought over for pharming? Kaiser J. Publication Types: News PMID: 18436771 [PubMed - indexed for MEDLINE] 185: Science. 2008 Apr 25;320(5875):425. Comment in: Science. 2008 Jul 25;321(5888):489. Seeds of a perfect storm. Fedoroff N. Publication Types: Editorial PMID: 18436745 [PubMed - indexed for MEDLINE] 186: Plant Biotechnol J. 2008 Aug;6(6):619-31. Epub 2008 Apr 22. Metabolic and genetic perturbations accompany the modification of galactomannan in seeds of Medicago truncatula expressing mannan synthase from guar (Cyamopsis tetragonoloba L.). Naoumkina M, Vaghchhipawala S, Tang Y, Ben Y, Powell RJ, Dixon RA. Plant Biology Division, Samuel Roberts Noble Foundation, 2510 Sam Noble Parkway, Ardmore, OK 73401, USA. Galactomannan gums are widely used in the food and oil industries, and there is considerable interest in applying biotechnological approaches to improve their physical properties. A mannan synthase from guar (Cyamopsis tetragonoloba) was expressed under the control of a bean beta-phaseolin promoter in transgenic Medicago truncatula. Although the expression of exogenous mannan synthase caused a slight decrease in galactomannan levels in Medicago, the molecular weight and viscosity of the polymer were significantly increased, although the mannose to galactose ratio and degree of polydispersity remained unchanged. At the same time, expression of about 2.8% of the genes was altered significantly in the seeds of transgenic Medicago lines analysed by Affymetrix genome chip, with a particularly striking induction of putative trehalose phosphate synthase genes. Mannan synthase expression also caused large alterations in the levels of a number of sugars and sugar alcohols, suggesting that over-expression of a processive glycosyltransferase perturbs the mechanisms of sugar sensing and/or homeostasis, possibly involving signalling via trehalose-6-phosphate. PMID: 18433421 [PubMed - indexed for MEDLINE] 187: J Nutr. 2008 May;138(5):921-6. Lysozyme transgenic goats' milk influences gastrointestinal morphology in young pigs. Brundige DR, Maga EA, Klasing KC, Murray JD. Department of Animal Science, University of California, Davis, CA 95616, USA. Transgenesis provides a method of expressing novel proteins in milk to increase the functional benefits of milk consumption. Transgenic goats expressing human lysozyme (hLZ) at 67% of the concentration in human breast milk were produced, thereby enhancing the antimicrobial properties of goats' milk. The objective of this study was to investigate the impact of pasteurized milk containing hLZ on growth, the intestinal epithelium, and an enteropathogenic Escherichia coli (EPEC) infection in young weaned pigs. Pigs were placed into 4 groups and fed a diet of solid food and either control (nontransgenic) goats' milk or milk from hLZ-transgenic goats. Growth was assessed by weight gain. Nonchallenged pigs were necropsied after 6 wk, whereas the remaining pigs were necropsied at 7 wk following bacterial challenge. We determined the numbers of total coliforms and E. coli and examined small intestinal histology for all pigs. Complete blood counts were also determined pre- and postchallenge. Challenged pigs receiving hLZ milk had fewer total coliforms (P = 0.029) and E. coli (P = 0.030) in the ileum than controls. hLZ-fed pigs also had a greater duodenal villi width (P = 0.029) than controls. Additionally, nonchallenged hLZ-fed pigs had fewer intraepithelial lymphocytes per micron of villi height (P = 0.020) than nonchallenged controls. These results indicate that the consumption of pasteurized hLZ goats' milk has the potential to improve gastrointestinal health and is protective against an EPEC in young weaned pigs. These same benefits may occur in young children if they were to consume milk from hLZ-transgenic goats. Publication Types: Research Support, Non-U.S. Gov't PMID: 18424602 [PubMed - indexed for MEDLINE] 188: FEBS Lett. 2008 May 14;582(11):1599-606. Epub 2008 Apr 16. Aggregation of proteins having Golgi apparatus sorting determinant induces large globular structures derived from the endoplasmic reticulum in plant seed cells. Maruyama N, Okuda E, Tatsuhara M, Utsumi S. Laboratory of Food Quality Design and Development, Graduate School of Agriculture, Kyoto University, Gokasho, Uji, Kyoto 611-0011, Japan. marunobu@kais.kyoto-u.ac.jp Endoplasmic reticulum (ER)-derived compartments are found in many plant species. Although it has been assumed that aggregation induces formation of the ER-derived compartments in plant seed cells, the effect of aggregation on the trafficking from the ER to the Golgi has not yet been elucidated. In this study, we used an aggregated type of red fluorescent protein (DsRED) to investigate the effect of aggregation on sorting in seed cells. DsRED fused to the Golgi sorting determinant was found mainly in large globular structures derived from the ER where ER-resident proteins were excluded. These results indicate that aggregation of the Golgi protein blocks transport from the ER to the Golgi. Publication Types: Research Support, Non-U.S. Gov't PMID: 18423406 [PubMed - indexed for MEDLINE] 189: Plant Biotechnol J. 2008 Jun;6(5):465-76. Epub 2008 Apr 14. Spatial and temporal expression of endosperm transfer cell-specific promoters in transgenic rice and barley. Li M, Singh R, Bazanova N, Milligan AS, Shirley N, Langridge P, Lopato S. Plant and Pest Science, School of Agriculture, Food and Wine, University of Adelaide, Waite Campus, Glen Osmond, SA 5064, Australia. Two putative endosperm-specific rice genes, OsPR602 and OsPR9a, were identified from database searches. The promoter regions of these genes were isolated, and transcriptional promoter:beta-glucuronidase (GUS) fusion constructs were stably transformed into rice and barley. The GUS expression patterns revealed that these promoters were active in early grain development in both rice and barley, and showed strongest expression in endosperm transfer cells during the early stages of grain filling. The GUS expression was similar in both rice and barley, but, in barley, expression was exclusively in the endosperm transfer cells and differed in timing of activation relative to rice. In rice, both promoters showed activity not only in the endosperm transfer cells, but also in the transfer cells of maternal tissue and in several floral tissues shortly before pollination. The expression patterns of OsPR602 and OsPR9a in flowers differed. The similarity of expression in both rice and barley suggests that these promoters may be useful to control transgene expression in the transfer cells of cereal grains with the aim of altering nutrient uptake or enhancing the barrier against pathogens at the boundary between maternal tissue and the developing endosperm. However, the expression during floral development should be considered if the promoters are used in rice. Publication Types: Research Support, Non-U.S. Gov't PMID: 18422887 [PubMed - indexed for MEDLINE] 190: FEMS Microbiol Lett. 2008 Jun;283(1):62-8. Epub 2008 Apr 16. Intragastric administration with recombinant Lactococcus lactis producing heme oxygenase-1 prevents lipopolysaccharide-induced endotoxemia in rats. Pang Q, Ji Y, Li Y, Bermúdez-Humarán LG, Hu G, Zeng Y. Department of Pharmacology, Nanjing Medical University, Nanjing, Jiangsu, China. Gut injury is a pivotal initiating event in the dysfunctional inflammatory response that causes postinjury multiple organ failure. Heme oxygenase-1 (HO-1) is an important enzyme that provides cellular protection against oxidative stress in different in vitro and in vivo systems. In this study, we evaluated the protective effects of intragastrically administered live Lactococcus lactis secreting bioactive HO-1 to treat intestinal mucosal injury induced by lipopolysaccharide in rats. Intragastric administration with this recombinant L. lactis strain led to active delivery of HO-1 at the mucosa and significantly decreased morbidity and mortality of lipopolysaccharide -induced endotoxemia as confirmed by blinded macroscopic and microscopic inflammatory scores (Chiu's grade), myeloperoxidase activity, mortality, and tumor necrosis factor-alpha and IL-10 cytokine stimulation. This protective effect could be abolished by an HO-1 inhibitor, the zinc protoporphyrin-IX. Our results suggest that a food-grade bacterium genetically modified to deliver bioactive HO-1 in situ exerts a protective effect against intestinal mucosal injury in rats with endotoxemia via modulation of the immune system. This novel approach may be beneficial for the maintenance of the intestinal barrier and anti-inflammatory response of the lower intestine. PMID: 18422629 [PubMed - indexed for MEDLINE] 191: Risk Anal. 2008 Apr;28(2):463-76. Farmer knowledge and risk analysis: postrelease evaluation of herbicide-tolerant canola in Western Canada. Mauro IJ, McLachlan SM. Environmental Conservation Lab, Department of Environment and Geography, University of Manitoba, Winnipeg, Canada. ian_mauro@umanitoba.ca The global controversy regarding the use of genetically modified (GM) crops has proved to be a challenge for "science-based" risk assessments. Although risk analysis incorporates societal perspectives in decision making over these crops, it is largely predicated on contrasts between "expert" and "lay" perspectives. The overall objective of this study is to explore the role for farmers' knowledge, and their decade-long experience with herbicide-tolerant (HT) canola, in the risk analysis of GM crops. From 2002 to 2003, data were collected using interviews (n= 15) and mail surveys (n= 370) with farmers from Manitoba and across Canada. The main benefits associated with HT canola were management oriented and included easier weed control, herbicide rotation, and better weed control, whereas the main risks were more diverse and included market harm, technology use agreements (TUAs), and increased seed costs. Benefits and risks were inversely related, and the salient factor influencing risk was farmer experiences with HT canola volunteers, followed by small farm size and duration using HT canola. These HT volunteers were reported by 38% of farmers, from both internal (e.g., seedbank, farm machinery, etc.) and external (e.g., wind, seed contamination, etc.) sources, and were found to persist over time. Farmer knowledge is a reliable and rich source of information regarding the efficacy of HT crops, demonstrating that individual experiences are important to risk perception. The socioeconomic nature of most risks combined with the continuing "farm income crisis" in North America demonstrates the need for a more holistic and inclusive approach to risk assessment associated with HT crops and, indeed, with all new agricultural technology. Publication Types: Research Support, Non-U.S. Gov't PMID: 18419662 [PubMed - indexed for MEDLINE] 192: Can J Physiol Pharmacol. 2008 Apr;86(4):215-21. Genetic possibilities for altering sunflower oil quality to obtain novel oils. Skorić D, Jocić S, Sakac Z, Lecić N. Serbian Academy of Sciences and Arts, Belgrade, Novi Sad Branch, Nikole Pasića 6, 21000 Novi Sad, Serbia. dragankoric@sbb.co.yu The sunflower is one of the four most important oilseed crops in the world, and the nutritional quality of its edible oil ranks among the best vegetable oils in cultivation. Typically up to 90% of the fatty acids in conventional sunflower oil are unsaturated, namely oleic (C 18:1, 16%-19%) and linoleic (C 18:2, 68%-72%) fatty acids. Palmitic (C 16:0, 6%), stearic (C 18:0, 5%), and minor amounts of myristic (C 14:0), myristoleic (C 14:1), palmitoleic (C 16:1), arachidic (C 20:0), behenic (C 22:0), and other fatty acids account for the remaining 10%. Advances in modern genetics, most importantly induced mutations, have altered the fatty acid composition of sunflower oil to a significant extent. Treating sunflower seeds with gamma- and X-rays has produced mutants with 25%-30% palmitic acid. Sunflower seed treatment with X-rays has also resulted in mutants having 30% palmitoleic acid, while treatments with mutagenic sodium azide have produced seeds containing 35% stearic acid. The most important mutations have been obtained by treatment with dimethyl sulfate, which produced genotypes with more than 90% oleic acid. Mutants have also been obtained that have a high linoleic acid content (>80%) by treating seeds with X-rays and ethyl methanesulfonate. Of the vitamin E family of compounds, sunflower oil is known to predominantly contain alpha-tocopherol (>90%). Spontaneous mutations controlled by recessive genes have been discovered that significantly alter tocopherol forms and levels. The genes in question are tph(1) (50% alpha- and 50% beta-tocopherol), tph(2) (0%-5% alpha- and 95%-100% gamma-tocopherol), and tph(1)tph(2) (8%-40% alpha-, 0%-25% beta-, 25%-84% gamma-, and 8%-50% delta-tocopherol). The existence of (mutant) genes for increased levels of individual fatty acids and for different forms and levels of tocopherol enables the development of sunflower hybrids with different oil quality. The greatest progress has been made in developing high-oleic hybrids (>90% oleic acid). There has been considerable work done recently on the development of high-oleic hybrids with altered tocopherol levels, the oil of which will have 10-20 times greater oxidative stability than that of conventional sunflower oil. While sunflower breeders work on developing hybrids with altered oil quality, medical scientists in general and nutritionists in particular will determine the parameters for the use of these novel types of oil that can improve human nutrition and be used in the prevention of cardiovascular diseases. Publication Types: Review PMID: 18418432 [PubMed - indexed for MEDLINE] 193: Transgenic Res. 2008 Dec;17(6):1059-77. Epub 2008 Apr 11. Environmental impact of herbicide regimes used with genetically modified herbicide-resistant maize. Devos Y, Cougnon M, Vergucht S, Bulcke R, Haesaert G, Steurbaut W, Reheul D. Department of Plant Production, Faculty of Bioscience Engineering, Ghent University, Coupure Links 653, 9000 Ghent, Belgium. Yann.Devos@UGent.be With the potential advent of genetically modified herbicide-resistant (GMHR) crops in the European Union, changes in patterns of herbicide use are predicted. Broad-spectrum, non-selective herbicides used with GMHR crops are expected to substitute for a set of currently used herbicides, which might alter the agro-environmental footprint from crop production. To test this hypothesis, the environmental impact of various herbicide regimes currently used with non-GMHR maize in Belgium was calculated and compared with that of possible herbicide regimes applied in GMHR maize. Impacts on human health and the environment were calculated through the pesticide occupational and environmental risk (POCER) indicator. Results showed that the environmental impact of herbicide regimes solely relying on the active ingredients glyphosate (GLY) or glufosinate-ammonium (GLU) is lower than that of herbicide regimes applied in non-GMHR maize. Due to the lower potential of GLY and GLU to contaminate ground water and their lower acute toxicity to aquatic organisms, the POCER exceedence factor values for the environment were reduced approximately by a sixth when GLY or GLU is used alone. However, the environmental impact of novel herbicide regimes tested may be underestimated due to the assumption that active ingredients used with GMHR maize would be used alone. Data retrieved from literature suggest that weed control efficacy is increased and resistance development delayed when GLY or GLU is used together with other herbicides in the GMHR system. Due to the partial instead of complete replacement of currently used herbicide regimes, the beneficial environmental impact of novel herbicide regimes might sometimes be reduced or counterbalanced. Despite the high weed control efficacy provided by the biotechnology-based weed management strategy, neither indirect harmful effects on farmland biodiversity through losses in food resources and shelter, nor shifts in weed communities have been demonstrated in GMHR maize yet. However, with the increasing adoption rate of GMHR maize and their associated novel herbicide regimes, this situation is expected to change in the short-term. PMID: 18404410 [PubMed - indexed for MEDLINE] 194: Food Chem Toxicol. 2008 Jun;46(6):2201-13. Epub 2008 Feb 29. Subchronic feeding study of herbicide-tolerant soybean DP-356Ø43-5 in Sprague-Dawley rats. Appenzeller LM, Munley SM, Hoban D, Sykes GP, Malley LA, Delaney B. Pioneer Hi-Bred International, Inc., Johnston, IA, USA. Optimum GAT1 soybean is a genetically modified (GM) soybean containing event DP-356Ø43-5 (356043) that was produced by integration of the coding sequences of the GAT4601 and GM-HRA proteins. In planta expression of these proteins confers tolerance to glyphosate and sulfonylurea/imidazolinone herbicides, respectively. This paper reports the results from a subchronic rat feeding study conducted with 356043 soybeans. Dehulled/defatted toasted meal and toasted ground hulls were prepared from soybeans from untreated plants (356043), herbicide-treated plants (356043+Gly/SU), non-transgenic isoline control (091), and three commercial non-transgenic reference varieties (93B86, 93B15, and 93M40). Individual diets conforming to standard certified rodent chow formulation (Purina Rodent LabDiet) 5002) were prepared with 20% meal (w/w) and 1.5% hulls (w/w). Diets were fed to young adult Sprague-Dawley rats (12/sex/group) for at least 93 days. Compared with rats fed the isoline control or conventional reference diets, no biologically-relevant, adverse effects were observed in rats fed diets containing 356043 or 356043+Gly/SU soybean with respect to body weight/gain, food consumption/efficiency, clinical signs, mortality, ophthalmology, neurobehavioral assessments (sensory response, grip strength, motor activity), clinical pathology (hematology, coagulation, serum chemistry, urinalysis), organ weights, and gross and microscopic pathology. The results from this study indicate that 356043 soybeans are as safe and nutritious as conventional non-GM soybeans. PMID: 18403083 [PubMed - indexed for MEDLINE] 195: Commun Agric Appl Biol Sci. 2007;72(4):805-11. Introduction a potato cultivar "sprit" as relatively resistant to main fungal pathogens causal agents of early blight and wilting on potato in Iran. Saremi H, Davoodvandy MH, Amarlou A. Department of plant pathology, Faculty of Agriculture Zanjan University, Iran. Potato (Solanum tubersum L.) is one of the most human food production cultured in Iran especially Zanjan province as a temperate region. Some fungal pathogens caused severely infected on potato tubers or foliage in the majority grown areas and resulted yield losses in potato production. Recent years from 2002 to 2004 infected samples were collected from different potato grown regions in Zanjan province then cultured on PDA after surface sterilization with sodium hypochlorite. Isolated fungal pathogens were identified and study showed the main pathogens with high incidence and frequency were Alternaria solani, Fusarium oxysporum and Verticillium sp. in studied areas. The regions which used convention varieties showed more diseases than other locations which used relatively resistant races. The rate of resistance for 10 international potato varieties was studied by inoculation of them by 10(5) spores suspension of three common fungal pathogens in the field. Study showed Sprit cultivar was more resistant than others to all three common pathogens and Lady-Claire was most susceptible. Yield production of Sprit per unit of land area was also exceeded that of other cultivars by factors of 1.10 to 2.25 respectively. The results of the study helped potato growers to culture Sprit cultivar and have good yield production in Zanjan and Hamedan provinces in this year. Publication Types: Research Support, Non-U.S. Gov't PMID: 18396814 [PubMed - indexed for MEDLINE] 196: Nature. 2008 Mar 6;452(7183):122-4. Almost in bloom. Marris E. PMID: 18396501 [PubMed - indexed for MEDLINE] 197: Curr Opin Biotechnol. 2008 Apr;19(2):181-9. Epub 2008 Apr 3. Metabolic engineering of plant volatiles. Dudareva N, Pichersky E. Department of Horticulture and Landscape Architecture, Purdue University, West Lafayette, IN 47907, USA. dudareva@purdue.edu Metabolic engineering of the volatile spectrum offers enormous potential for plant improvement because of the great contribution of volatile secondary metabolites to reproduction, defense and food quality. Recent advances in the identification of the genes and enzymes responsible for the biosynthesis of volatile compounds have made this metabolic engineering highly feasible. Notable successes have been reported in enhancing plant defenses and improving scent and aroma quality of flowers and fruits. These studies have also revealed challenges and limitations which will be likely surmounted as our understanding of plant volatile network improves. Publication Types: Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Review PMID: 18394878 [PubMed - indexed for MEDLINE] 198: J Proteome Res. 2008 May;7(5):1850-61. Epub 2008 Apr 5. Proteomics as a complementary tool for identifying unintended side effects occurring in transgenic maize seeds as a result of genetic modifications. Zolla L, Rinalducci S, Antonioli P, Righetti PG. Department of Environmental Sciences, University of Tuscia, Viterbo, Italy. zolla@unitus.it To improve the probability of detecting unintended side effects during maize gene manipulations by bombardment, proteomics was used as an analytical tool complementary to the existing safety assessment techniques. Since seed proteome is highly dynamic, depending on the species variability and environmental influence, we analyzed the proteomic profiles of one transgenic maize variety (event MON 810) in two subsequent generations (T05 and T06) with their respective isogenic controls (WT05 and WT06). Thus, by comparing the proteomic profiles of WT05 with WT06 we could determine the environmental effects, while the comparison between WT06 and T06 seeds from plants grown under controlled conditions enabled us to investigate the effects of DNA manipulation. Finally, by comparison of T05 with T06 seed proteomes, it was possible to get some indications about similarities and differences between the adaptations of transgenic and isogenic plants to the same strictly controlled growth environment. Approximately 100 total proteins resulted differentially modulated in the expression level as a consequence of the environmental influence (WT06 vs WT05), whereas 43 proteins resulted up- or down-regulated in transgenic seeds with respect to their controls (T06 vs WT06), which could be specifically related to the insertion of a single gene into a maize genome by particle bombardment. Transgenic seeds responded differentially to the same environment as compared to their respective isogenic controls, as a result of the genome rearrangement derived from gene insertion. To conclude, an exhaustive differential proteomic analysis allows to determine similarities and differences between traditional food and new products (substantial equivalence), and a case-by-case assessment of the new food should be carried out in order to have a wide knowledge of its features. Publication Types: Research Support, Non-U.S. Gov't PMID: 18393457 [PubMed - indexed for MEDLINE] 199: Nat Biotechnol. 2008 Apr;26(4):379; discussion 379-80. Comment on: Nat Biotechnol. 2007 Dec;25(12):1330. An inconvenient version of events. Monastra G. Publication Types: Comment Letter PMID: 18392011 [PubMed - indexed for MEDLINE] 200: Nat Biotechnol. 2008 Apr;26(4):365. Tear-free onions. Aldridge S. Publication Types: News PMID: 18392004 [PubMed - indexed for MEDLINE] 201: Br J Nutr. 2008 Feb;99 Suppl 1:S22-5. Influence of parental attitudes in the development of children eating behaviour. Scaglioni S, Salvioni M, Galimberti C. Pediatric Clinic S. Paolo Hospital University of Milan, Milan, Italy. silviascaglioni@unimi.it The present paper is a review of available data on effects of parental feeding attitudes and styles on child nutritional behaviour. Food preferences develop from genetically determined predispositions to like sweet and salty flavours and to dislike bitter and sour tastes. There is evidence for existence of some innate, automatic mechanism that regulate appetite. However, from birth genetic predispositions are modified by experience. There are mechanisms of taste development: mere exposure, medicine effect, flavour learning, flavour nutrient learning. Parents play a pivotal role in the development of their child's food preferences and energy intake, with research indicating that certain child feeding practices, such as exerting excessive control over what and how much children eat, may contribute to childhood overweight. Mothers are of particular interest on children's eating behaviour, as they have been shown to spend significantly more time than fathers in direct interactions with their children across several familial situations.A recent paper describes two primary aspects of control: restriction, which involves restricting children's access to junk foods and restricting the total amount of food, and pressure, which involves pressuring children to eat healthy foods (usually fruits and vegetables) and pressuring to eat more in general.The results showed significant correlations between parent and child for reported nutritional behaviour like food intake, eating motivations, and body dis- and satisfaction. Parents create environments for children that may foster the development of healthy eating behaviours and weight, or that may promote overweight and aspects of disordered eating. In conclusion positive parental role model may be a better method for improving a child's diet than attempts at dietary control. Publication Types: Review PMID: 18257948 [PubMed - indexed for MEDLINE] 202: Curr Opin Biotechnol. 2008 Apr;19(2):129-30. Epub 2008 Apr 2. Plant biotechnology--predictive, green and quantitative. Chappell J, Grotewold E. Publication Types: Editorial PMID: 18387798 [PubMed - indexed for MEDLINE] 203: Environ Biosafety Res. 2008 Jan-Mar;7(1):35-56. Epub 2008 Apr 3. A screening method for prioritizing non-target invertebrates for improved biosafety testing of transgenic crops. Todd JH, Ramankutty P, Barraclough EI, Malone LA. The Horticulture and Food Research Institute of New Zealand Limited (HortResearch), Mt Albert, Private Bag 92169, Auckland Mail Centre, Auckland 1142, New Zealand. jtodd@hortresearch.co.nz We have developed a screening method that can be used during the problem formulation phase of risk assessment to identify and prioritize non-target invertebrates for risk analysis with any transgenic plant. In previously published protocols for this task, five criteria predominated. These criteria have been combined by our method in a simple model which assesses: (1) the possible level of risk presented by the plant to each invertebrate species (through measurements of potential hazard and exposure, the two principal criteria); (2) the hypothetical environmental impact of this risk (determined by the currently known status of the species' population in the ecosystem and its potential resilience to environmental perturbations); (3) the estimated economic, social and cultural value of each species; and (4) the assessed ability to conduct tests with the species. The screening method uses information on each of these criteria entered into a specially designed database that was developed using Microsoft Access 2003. The database holds biological and ecological information for each non-target species, as well as information about the transgenic plant that is the subject of the risk assessment procedure. Each piece of information is then ranked on the basis of the value of the information to each criterion being measured. This ranking system is flexible, allowing the method to be easily adapted for use in any agro-ecosystem and with any plant modification. A model is then used to produce a Priority Ranking of Non-Target Invertebrates (PRONTI) score for each species, which in turn allows the species to be prioritized for risk assessment. As an example, the method was used to prioritize non-target invertebrates for risk assessment of a hypothetical introduction of Bacillus thuringiensis (Bt) Cry1Ac-expressing Pinus radiata trees into New Zealand. Publication Types: Research Support, Non-U.S. Gov't PMID: 18384728 [PubMed - indexed for MEDLINE] 204: Rev Med Suisse. 2008 Jan 30;4(142):314. [Food and milk clones declared] [Article in French] Nau JY. PMID: 18383942 [PubMed - indexed for MEDLINE] 205: Food Chem Toxicol. 2008 Jun;46(6):1994-2002. Epub 2008 Feb 2. Comparison of grain from corn rootworm resistant transgenic DAS-59122-7 maize with non-transgenic maize grain in a 90-day feeding study in Sprague-Dawley rats. He XY, Huang KL, Li X, Qin W, Delaney B, Luo YB. College of Food Science and Nutritional Engineering, China Agricultural University, No. 17 Tsinghua Donglu, Beijing 100083, China. DAS-59122-7 (59122) is a transgenic maize (Zea mays L.) that contains genes encoding Cry34Ab1 and Cry35Ab1 proteins from Bacillus thuringiensis Berliner strain 149B1 and phosphinothricin acetyltransferase (PAT) protein from Streptomyces viridochromogenes. Expression of these proteins in planta confers resistance to corn rootworms and other Coleopteran parasites and tolerance to herbicides containing glufosinate ammonium, respectively. In the current study, processed flours from 59122 maize grain or its near isogenic control line (091) were used at two concentrations (50% and 70% wt/wt) to produce diets that were fed to rats for 90 days in accordance with Chinese toxicology guidelines (GB15193.13-2003). A commercial AIN93G diet was used as an additional negative control. No significant differences in body weight and feed utilization were observed between rats consuming diets formulated with 59122 and 091 Control corn. Statistical differences (p<0.05) were observed in certain hematology and serum chemistry response variables between rats consuming diets formulated with 59122 or 091 Control flour compared to AIN93G diet. However, the mean value of these response variables in the 59122 groups were not statistically different from those observed in diets formulated with corresponding high and low concentrations of the flour from the 091 Control maize grain. Therefore, the statistical differences were considered to be related to consumption of diets containing high concentrations of maize flour (compared to AIN93G diets) regardless of source rather than to consumption of flour from 59122 maize grain. The results from this study demonstrated that 59122 maize grain is as safe as non-transgenic maize grain. Publication Types: Research Support, Non-U.S. Gov't PMID: 18381227 [PubMed - indexed for MEDLINE] 206: J Am Vet Med Assoc. 2008 Mar 1;232(5):667. FDA reaches final conclusion that food from clones is safe. [No authors listed] Publication Types: News PMID: 18380061 [PubMed - indexed for MEDLINE] 207: J Exp Bot. 2008;59(6):1305-13. Epub 2008 Mar 28. Histological characterization of root-knot nematode resistance in cowpea and its relation to reactive oxygen species modulation. Das S, DeMason DA, Ehlers JD, Close TJ, Roberts PA. Department of Botany and Plant Sciences, University of California, Riverside, CA 92521, USA. Root-knot nematodes (Meloidogyne spp.) are sedentary endoparasites with a broad host range which includes economically important crop species. Cowpea (Vigna unguiculata L. Walp) is an important food and fodder legume grown in many regions where root-knot nematodes are a major problem in production fields. Several sources of resistance to root-knot nematode have been identified in cowpea, including the widely used Rk gene. As part of a study to elucidate the mechanism of Rk-mediated resistance, the histological response to avirulent M. incognita feeding of a resistant cowpea cultivar CB46 was compared with a susceptible near-isogenic line (in CB46 background). Most root-knot nematode resistance mechanisms in host plants that have been examined induced a hypersensitive response (HR). However, there was no typical HR in resistant cowpea roots and nematodes were able to develop normal feeding sites similar to those in susceptible roots up to 9-14 d post inoculation (dpi). From 14-21 dpi giant cell deterioration was observed and the female nematodes showed arrested development and deterioration. Nematodes failed to reach maturity and did not initiate egg laying in resistant roots. These results confirmed that the induction of resistance is relatively late in this system. Typically in pathogen resistance HR is closely associated with an oxidative burst (OB) in infected tissue. The level of reactive oxygen species release in both compatible and incompatible reactions during early and late stages of infection was also quantified. Following a basal OB during early infection in both susceptible and resistant roots, which was also observed in mechanically wounded root tissues, no significant OB was detected up to 14 dpi, a profile consistent with the histological observations of a delayed resistance response. These results will be useful to design gene expression experiments to dissect Rk-mediated resistance at the molecular level. Publication Types: Research Support, U.S. Gov't, Non-P.H.S. PMID: 18375605 [PubMed - indexed for MEDLINE] 208: Nature. 2008 Mar 20;452(7185):273-7. Comment in: Nature. 2008 May 29;453(7195):587. Water: more crop per drop. Marris E. Publication Types: News PMID: 18354452 [PubMed - indexed for MEDLINE] 209: J Neurosci. 2008 Mar 19;28(12):3103-13. Rapid consolidation to a radish and protein synthesis-dependent long-term memory after single-session appetitive olfactory conditioning in Drosophila. Krashes MJ, Waddell S. Department of Neurobiology, University of Massachusetts Medical School, Worcester, Massachusetts 01605, USA. In Drosophila, formation of aversive olfactory long-term memory (LTM) requires multiple training sessions pairing odor and electric shock punishment with rest intervals. In contrast, here we show that a single 2 min training session pairing odor with a more ethologically relevant sugar reinforcement forms long-term appetitive memory that lasts for days. Appetitive LTM has some mechanistic similarity to aversive LTM in that it can be disrupted by cycloheximide, the dCreb2-b transcriptional repressor, and the crammer and tequila LTM-specific mutations. However, appetitive LTM is completely disrupted by the radish mutation that apparently represents a distinct mechanistic phase of consolidated aversive memory. Furthermore, appetitive LTM requires activity in the dorsal paired medial neuron and mushroom body alpha'beta' neuron circuit during the first hour after training and mushroom body alphabeta neuron output during retrieval, suggesting that appetitive middle-term memory and LTM are mechanistically linked. Last, experiments feeding and/or starving flies after training reveals a critical motivational drive that enables appetitive LTM retrieval. Publication Types: Research Support, N.I.H., Extramural PMID: 18354013 [PubMed - indexed for MEDLINE] 210: PLoS ONE. 2008 Mar 19;3(3):e1818. A built-in strategy for containment of transgenic plants: creation of selectively terminable transgenic rice. Lin C, Fang J, Xu X, Zhao T, Cheng J, Tu J, Ye G, Shen Z. Institute of Insect Sciences and State Key Laboratory of Rice Biology, Zhejiang University, Hangzhou, China. Plant transgenic technology has been widely utilized for engineering crops for trait improvements and for production of high value proteins such as pharmaceuticals. However, the unintended spreading of commercial transgenic crops by pollination and seed dispersal is a major concern for environmental and food safety. Simple and reliable containment strategies for transgenes are highly desirable. Here we report a novel method for creating selectively terminable transgenic rice. In this method, the gene(s) of interest is tagged with a RNA interference cassette, which specifically suppresses the expression of the bentazon detoxification enzyme CYP81A6 and thus renders transgenic rice to be sensitive to bentazon, a herbicide used for rice weed control. We generated transgenic rice plants by this method using a new glyphosate resistant 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) gene from Pesudomonas putida as the gene of interest, and demonstrated that these transgenic rice plants were highly sensitive to bentazon but tolerant to glyphosate, which is exactly the opposite of conventional rice. Field trial of these transgenic rice plants further confirmed that they can be selectively killed at 100% by one spray of bentazon at a regular dose used for conventional rice weed control. Furthermore, we found that the terminable transgenic rice created in this study shows no difference in growth, development and yield compared to its non-transgenic control. Therefore, this method of creating transgenic rice constitutes a novel strategy of transgene containment, which appears simple, reliable and inexpensive for implementation. Publication Types: Research Support, Non-U.S. Gov't PMID: 18350155 [PubMed - indexed for MEDLINE] 211: Food Chem Toxicol. 2008 May;46 Suppl 2:S71-97. Epub 2008 Feb 2. Evaluation of protein safety in the context of agricultural biotechnology. Delaney B, Astwood JD, Cunny H, Conn RE, Herouet-Guicheney C, Macintosh S, Meyer LS, Privalle L, Gao Y, Mattsson J, Levine M; ILSI International Food Biotechnology Committee Task Force on Protein Safety. Collaborators: Privalle L, Zhou J, Eichen-Conn R, Juberg D, Bannon G, Delaney B, Meyer L. Pioneer Hi-Bred, A DuPont Company, Johnston, IA, USA. One component of the safety assessment of agricultural products produced through biotechnology is evaluation of the safety of newly expressed proteins. The ILSI International Food Biotechnology Committee has developed a scientifically based two-tiered, weight-of-evidence strategy to assess the safety of novel proteins used in the context of agricultural biotechnology. Recommendations draw upon knowledge of the biological and chemical characteristics of proteins and testing methods for evaluating potential intrinsic hazards of chemicals. Tier I (potential hazard identification) includes an assessment of the biological function or mode of action and intended application of the protein, history of safe use, comparison of the amino acid sequence of the protein to other proteins, as well as the biochemical and physico-chemical properties of the proteins. Studies outlined in Tier II (hazard characterization) are conducted when the results from Tier I are not sufficient to allow a determination of safety (reasonable certainty of no harm) on a case-by-case basis. These studies may include acute and repeated dose toxicology studies and hypothesis-based testing. The application of these guidelines is presented using examples of transgenic proteins applied for agricultural input and output traits in genetically modified crops along with recommendations for future research considerations related to protein safety assessment. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 18348900 [PubMed - indexed for MEDLINE] 212: Environ Entomol. 2008 Feb;37(1):263-70. Tri-trophic interactions between Bt cotton, the herbivore Aphis gossypii Glover (Homoptera: Aphididae), and the predator Chrysopa pallens (Rambur) (Neuroptera: Chrysopidae). Guo JY, Wan FH, Dong L, Lövei GL, Han ZJ. State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100094, China. guojy@cjac.org.cn Tri-trophic impacts of transgenic Bacillus thuringiensis (Bt) cotton GK12 and NuCOTN 99B were studied using a predator, the great lacewing Chrysopa pallens (Rambur), and its prey, the cotton aphid Aphis gossypii Glover, in laboratory feeding experiments. The parental nontransgenic cotton cultivar of GK12 was used as control. The predator was fed with uniform (aphids from a single cultivar) or mixed prey (aphids from the three cotton cultivars provided on alternate days). Mortality and development of the immature stages, pupal body mass, adult sex ratio, fecundity, and egg viability of C. pallens were measured. When fed GK12-originated aphid prey, pupal body mass of C. pallens was significantly higher than that of the control, more females emerged, and these females laid significantly more eggs. Other parameters were not impacted. Females emerging from larvae maintained on NuCOTN 99B-originated prey laid fewer eggs than those maintained on GK12. Other measurements did not differ significantly between the two Bt cotton cultivars. Compared with the control, mixed feeding significantly prolonged pupal development time and increased pupal body mass and percentage of females but did not affect other parameters. These results indicate that C. pallens is sensitive to aphid prey from different cotton cultivars. Transgenic Bt cotton GK12-originated aphid prey has no adverse impact on survival, development, and fecundity of C. pallens. Between the two Bt cotton cultivars, NuCOTN 99B-originated aphid prey provided to C. pallens in the larval stage may lower female fecundity. Mixed feeding of C. pallens with the two Bt cotton-originated prey and non-Bt prey may have some adverse impacts on pupal development. Publication Types: Comparative Study Research Support, Non-U.S. Gov't PMID: 18348819 [PubMed - indexed for MEDLINE] 213: Arch Environ Contam Toxicol. 2008 Nov;55(4):584-92. Epub 2008 Mar 18. Reduced fitness of Daphnia magna fed a Bt-transgenic maize variety. Bøhn T, Primicerio R, Hessen DO, Traavik T. Genøk-Centre for Biosafety, The Science Park, P.O. Box 6418, Tromso, 9294, Norway. thomas@genok.org Genetically modified (GM) maize expressing the Bt-toxin Cry1Ab (Bt-maize) was tested for effects on survival, growth, and reproduction of the water flea Daphnia magna, a crustacean arthropod commonly used as a model organism in ecotoxicological studies. In three repeated experiments, D. magna were fed 100% ground maize in suspension, using either GM or isogenic unmodified (UM) maize. D. magna fed GM-maize showed a significantly reduced fitness performance: The mortality was higher, a lower proportion of females reached sexual maturation, and the overall egg production was lower compared to D. magna fed UM isogenic maize. We conclude that the tested variety of Bt-maize and its UM counterpart do not have the same quality as food sources for this widely used model organism. The combination of a reduced fitness performance combined with earlier onset of reproduction of D. magna fed Bt-maize indicates a toxic effect rather than a lower nutritional value of the GM-maize. Publication Types: Research Support, Non-U.S. Gov't PMID: 18347840 [PubMed - indexed for MEDLINE] 214: Plant Biotechnol J. 2008 May;6(4):403-15. Epub 2008 Mar 12. Interlinking showy traits: co-engineering of scent and colour biosynthesis in flowers. Ben Zvi MM, Negre-Zakharov F, Masci T, Ovadis M, Shklarman E, Ben-Meir H, Tzfira T, Dudareva N, Vainstein A. The Institute of Plant Sciences and Genetics in Agriculture, Faculty of Agricultural, Food and Environmental Quality Sciences, The Hebrew University of Jerusalem, PO Box 12, Rehovot 76100, Israel. The phenylpropanoid pathway gives rise to metabolites that determine floral colour and fragrance. These metabolites are one of the main means used by plants to attract pollinators, thereby ensuring plant survival. A lack of knowledge about factors regulating scent production has prevented the successful enhancement of volatile phenylpropanoid production in flowers. In this study, the Production of Anthocyanin Pigment1 (Pap1) Myb transcription factor from Arabidopsis thaliana, known to regulate the production of non-volatile phenylpropanoids, including anthocyanins, was stably introduced into Petunia hybrida. In addition to an increase in pigmentation, Pap1-transgenic petunia flowers demonstrated an increase of up to tenfold in the production of volatile phenylpropanoid/benzenoid compounds. The dramatic increase in volatile production corresponded to the native nocturnal rhythms of volatile production in petunia. The application of phenylalanine to Pap1-transgenic flowers led to an increase in the otherwise negligible levels of volatiles emitted during the day to nocturnal levels. On the basis of gene expression profiling and the levels of pathway intermediates, it is proposed that both increased metabolic flux and transcriptional activation of scent and colour genes underlie the enhancement of petunia flower colour and scent production by Pap1. The co-ordinated regulation of metabolic steps within or between pathways involved in vital plant functions, as shown here for two showy traits determining plant-pollinator interactions, provides a clear advantage for plant survival. The use of a regulatory factor that activates scent production creates a new biotechnological strategy for the metabolic architecture of fragrance, leading to the creation of novel genetic variability for breeding purposes. Publication Types: Research Support, Non-U.S. Gov't PMID: 18346094 [PubMed - indexed for MEDLINE] 215: Medicina (Kaunas). 2008;44(2):87-99. Genetically modified organisms: do the benefits outweigh the risks? Hug K. K. Hug, Department of Medical Ethics, Lund University, BMC C 13, 221 84 Lund, Sweden. Kristina.Hug@med.lu.se The objective of this literature review is to analyze the implications of using genetically modified organisms (GMOs) as well as international and European position regarding such organisms. METHOD: Review of international and European legal requirements and ethical guidelines and relevant publications, found and accessed with the help of PubMed and Lund University Library databases. RESULTS: The article discusses the main application areas of GMOs, the expansion of using GMOs in the world as well as the advantages and disadvantages of the implications of their usage. It further provides an overview of the suggested ways to tackle or avoid the GMO-related risks. The international and European positions regarding the application of GMOs are discussed and European Directives, Regulations, and ethical guidelines are overviewed. The article further presents the public attitudes towards GMOs in Europe as well as overviews surveys conducted at the national level. CONCLUSION: Suggested steps to tackle the challenge of developing and managing biotechnology for the benefit of public health and the environment are presented. Publication Types: Comparative Study Review PMID: 18344661 [PubMed - indexed for MEDLINE] 216: Shokuhin Eiseigaku Zasshi. 2008 Feb;49(1):16-22. Development of event-specific quantitation method for GA21 maize, which is a gm event without CaMV35S promoter. Oguchi T, Onishi M, Chikagawa Y, Minegishi Y, Kodama T, Akiyama H, Ohno Y, Futo S, Hino A, Furui S, Kitta K. National Agriculture and Food Research Organization, National Food Research Institute, Ibaraki, Japan. A real-time PCR detection method was developed for event-specific quantitation of Roundup Ready maize, GA21. The developed PCR method was designed to amplify an artificial junction site between the native maize genome DNA and the recombinant DNA of GA21 maize, which provides only one target sequence per haploid of GA21 genome. Thus, the amplification efficiency of the event-specific target for GA21 became closely similar to the amplification of SSIIb, and the conversion factor (Cf) for the quantitation method was similar to the theoretical value. The developed method demonstrated better performance than the existing construct-specific method that has been used as a Japanese official method. The developed method can easily be combined with the real-time PCR targeting of the CaMV35S promoter, and the multiplexed method should be an effective screening method for GM maize. Publication Types: Research Support, Non-U.S. Gov't PMID: 18344654 [PubMed - indexed for MEDLINE] 217: Science. 2008 Mar 14;319(5869):1474-6. Erratum in: Science. 2008 Apr 4;320(5872):50. Science.2008 Apr 18;320(5874):316. Agriculture. Dueling visions for a hungry world. Stokstad E. Publication Types: News PMID: 18339914 [PubMed - indexed for MEDLINE] 218: Appetite. 2008 Jul;51(1):129-36. Epub 2008 Feb 7. Acceptance of genetically modified foods: the relation between technology and evaluation. Tenbült P, De Vries NK, van Breukelen G, Dreezens E, Martijn C. Department of Health Education and Promotion, Universiteit Maastricht, The Netherlands. p.m.a.tenbult@uvt.nl This study investigates why consumers accept different genetically modified food products to different extents. The study shows that whether food products are genetically modified or not and whether they are processed or not are the two important features that affect the acceptance of food products and their evaluation (in terms of perceived healthiness, naturalness, necessity and tastiness). The extent to which these evaluation attributes and acceptance of a product are affected by genetic modification or processing depends on whether the product is negatively affected by the other technology: Any technological change to a 'natural' product (when nonprocessed products are genetically modified or when non-genetically modified products are processed) affect evaluation and acceptance stronger than a change to an technologically adapted product (when processed products are also genetically modified or vice versa). Furthermore, evaluation attributes appear to mediate the effects of genetic modification and processing on acceptance. Publication Types: Randomized Controlled Trial PMID: 18336952 [PubMed - indexed for MEDLINE] 219: Food Chem Toxicol. 2008 Mar;46 Suppl 1:S2-70. Epub 2008 Feb 13. Safety and nutritional assessment of GM plants and derived food and feed: the role of animal feeding trials. EFSA GMO Panel Working Group on Animal Feeding Trials. Collaborators: Alink G, Barlow S, Cockburn A, Flachowsky G, Knudsen I, Kuiper H, Massin DP, Pascal G, Peijnenburg A, Phipps R, Pöting A, Poulsen M, Seinen W, Spielmann H, van Loveren H, Wal JM, Williams A, Andersson HC, Arpaia S, Bartsch D, Casacuberta J, Davies H, De Loose M, Hendriksen N, Herman L, Kärenlampi S, Kiss J, Kryspin-Sørensen I, Kuiper H, Nes I, Panopoulos N, Perry J, Pöting A, Schiemann J, Seinen W, Sweet J, Wal JM. In this report the various elements of the safety and nutritional assessment procedure for genetically modified (GM) plant derived food and feed are discussed, in particular the potential and limitations of animal feeding trials for the safety and nutritional testing of whole GM food and feed. The general principles for the risk assessment of GM plants and derived food and feed are followed, as described in the EFSA guidance document of the EFSA Scientific Panel on Genetically Modified Organisms. In Section 1 the mandate, scope and general principles for risk assessment of GM plant derived food and feed are discussed. Products under consideration are food and feed derived from GM plants, such as maize, soybeans, oilseed rape and cotton, modified through the introduction of one or more genes coding for agronomic input traits like herbicide tolerance and/or insect resistance. Furthermore GM plant derived food and feed, which have been obtained through extensive genetic modifications targeted at specific alterations of metabolic pathways leading to improved nutritional and/or health characteristics, such as rice containing beta-carotene, soybeans with enhanced oleic acid content, or tomato with increased concentration of flavonoids, are considered. The safety assessment of GM plants and derived food and feed follows a comparative approach, i.e. the food and feed are compared with their non-GM counterparts in order to identify intended and unintended (unexpected) differences which subsequently are assessed with respect to their potential impact on the environment, safety for humans and animals, and nutritional quality. Key elements of the assessment procedure are the molecular, compositional, phenotypic and agronomic analysis in order to identify similarities and differences between the GM plant and its near isogenic counterpart. The safety assessment is focussed on (i) the presence and characteristics of newly expressed proteins and other new constituents and possible changes in the level of natural constituents beyond normal variation, and on the characteristics of the GM food and feed, and (ii) the possible occurrence of unintended (unexpected) effects in GM plants due to genetic modification. In order to identify these effects a comparative phenotypic and molecular analysis of the GM plant and its near isogenic counterpart is carried out, in parallel with a targeted analysis of single specific compounds, which represent important metabolic pathways in the plant like macro and micro nutrients, known anti-nutrients and toxins. Significant differences may be indicative of the occurrence of unintended effects, which require further investigation. Section 2 provides an overview of studies performed for the safety and nutritional assessment of whole food and feed. Extensive experience has been built up in recent decades from the safety and nutritional testing in animals of irradiated foods, novel foods and fruit and vegetables. These approaches are also relevant for the safety and nutritional testing of whole GM food and feed. Many feeding trials have been reported in which GM foods like maize, potatoes, rice, soybeans and tomatoes have been fed to rats or mice for prolonged periods, and parameters such as body weight, feed consumption, blood chemistry, organ weights, histopathology etc have been measured. The food and feed under investigation were derived from GM plants with improved agronomic characteristics like herbicide tolerance and/or insect resistance. The majority of these experiments did not indicate clinical effects or histopathological abnormalities in organs or tissues of exposed animals. In some cases adverse effects were noted, which were difficult to interpret due to shortcomings in the studies. Many studies have also been carried out with feed derived from GM plants with agronomic input traits in target animal species to assess the nutritive value of the feed and their performance potential. Studies in sheep, pigs, broilers, lactating dairy cows, and fish, comparing the in vivo bioavailability of nutrients from a range of GM plants with their near isogenic counterpart and commercial varieties, showed that they were comparable with those for near isogenic non-GM lines and commercial varieties. In Section 3 toxicological in vivo, in silico, and in vitro test methods are discussed which may be applied for the safety and nutritional assessment of specific compounds present in food and feed or of whole food and feed derived from GM plants. Moreover the purpose, potential and limitations of the 90-day rodent feeding trial for the safety and nutritional testing of whole food and feed have been examined. Methods for single and repeated dose toxicity testing, reproductive and developmental toxicity testing and immunotoxicity testing, as described in OECD guideline tests for single well-defined chemicals are discussed and considered to be adequate for the safety testing of single substances including new products in GM food and feed. Various in silico and in vitro methods may contribute to the safety assessment of GM plant derived food and feed and components thereof, like (i) in silico searches for sequence homology and/or structural similarity of novel proteins or their degradation products to known toxic or allergenic proteins, (ii) simulated gastric and intestinal fluids in order to study the digestive stability of newly expressed proteins and in vitro systems for analysis of the stability of the novel protein under heat or other processing conditions, and (iii) in vitro genotoxicity test methods that screen for point mutations, chromosomal aberrations and DNA damage/repair. The current performance of the safety assessment of whole foods is mainly based on the protocols for low-molecular-weight chemicals such as pharmaceuticals, industrial chemicals, pesticides, food additives and contaminants. However without adaptation, these protocols have limitations for testing of whole food and feed. This primarily results from the fact that defined single substances can be dosed to laboratory animals at very large multiples of the expected human exposure, thus giving a large margin of safety. In contrast foodstuffs are bulky, lead to satiation and can only be included in the diet at much lower multiples of expected human intakes. When testing whole foods, the possible highest concentration of the GM food and feed in the laboratory animal diet may be limited because of nutritional imbalance of the diet, or by the presence of compounds with a known toxicological profile. The aim of the 90-days rodent feeding study with the whole GM food and feed is to assess potential unintended effects of toxicological and/or nutritional relevance and to establish whether the GM food and feed is as safe and nutritious as its traditional comparator rather than determining qualitative and quantitative intrinsic toxicity of defined food constituents. The design of the study should be adapted from the OECD 90-day rodent toxicity study. The precise study design has to take into account the nature of the food and feed and the characteristics of the new trait(s) and their intended role in the GM food and feed. A 90-day animal feeding trial has a large capacity (sensitivity and specificity) to detect potential toxicological effects of single well defined compounds. This can be concluded from data reported on the toxicology of a wide range of industrial chemicals, pharmaceuticals, food substances, environmental, and agricultural chemicals. It is possible to model the sensitivity of the rat subchronic feeding study for the detection of hypothetically increased amount of compounds such as anti-nutrients, toxicants or secondary metabolites. With respect to the detection of potential unintended effects in whole GM food and feed, it is unlikely that substances present in small amounts and with a low toxic potential will result in any observable (unintended) effects in a 90-day rodent feeding study, as they would be below the no-observed-effect-level and thus of unlikely impact to human health at normal intake levels. Laboratory animal feeding studies of 90-days duration appear to be sufficient to pick up adverse effects of diverse compounds that would also give adverse effects after chronic exposure. This conclusion is based on literature data from studies investigating whether toxicological effects are adequately identified in 3-month subchronic studies in rodents, by comparing findings at 3 and 24 months for a range of different chemicals. The 90-day rodent feeding study is not designed to detect effects on reproduction or development other than effects on adult reproductive organ weights and histopathology. Analyses of available data indicate that, for a wide range of substances, reproductive and developmental effects are not potentially more sensitive endpoints than those examined in subchronic toxicity tests. Should there be structural alerts for reproductive/developmental effects or other indications from data available on a GM food and feed, then these tests should be considered. By relating the estimated daily intake, or theoretical maximum daily intake per capita for a given whole food (or the sum of its individual commercial constituents) to that consumed on average per rat per day in the subchronic 90-day feeding study, it is possible to establish the margin of exposure (safety margin) for consumers. Results obtained from testing GM food and feed in rodents indicate that large (at least 100-fold) 'safety' margins exist between animal exposure levels without observed adverse effects and estimated human daily intake. Results of feeding studies with feed derived from GM plants with improved agronomic properties, carried out in a wide range of livestock species, are discussed. The studies did not show any biologically relevant differences in the parameters tested between control and test animals. (ABSTRACT TRUNCATED) Publication Types: Review PMID: 18328408 [PubMed - indexed for MEDLINE] 220: Plant Cell Rep. 2008 Jun;27(6):1027-38. Epub 2008 Mar 8. Agrobacterium tumefaciens-mediated transformation of poinsettia, Euphorbia pulcherrima, with virus-derived hairpin RNA constructs confers resistance to Poinsettia mosaic virus. Clarke JL, Spetz C, Haugslien S, Xing S, Dees MW, Moe R, Blystad DR. Plant Health and Plant Protection Division, Norwegian Institute for Agricultural and Environmental Research, Hoegskoleveien 7, 1432 Aas, Norway. jihong.liu-clarke@bioforsk.no Agrobacterium-mediated transformation for poinsettia (Euphorbia pulcherrima Willd. Ex Klotzsch) is reported here for the first time. Internode stem explants of poinsettia cv. Millenium were transformed by Agrobacterium tumefaciens, strain LBA 4404, harbouring virus-derived hairpin (hp) RNA gene constructs to induce RNA silencing-mediated resistance to Poinsettia mosaic virus (PnMV). Prior to transformation, an efficient somatic embryogenesis system was developed for poinsettia cv. Millenium in which about 75% of the explants produced somatic embryos. In 5 experiments utilizing 868 explants, 18 independent transgenic lines were generated. An average transformation frequency of 2.1% (range 1.2-3.5%) was revealed. Stable integration of transgenes into the poinsettia nuclear genome was confirmed by PCR and Southern blot analysis. Both single- and multiple-copy transgene integration into the poinsettia genome were found among transformants. Transgenic poinsettia plants showing resistance to mechanical inoculation of PnMV were detected by double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA). Northern blot analysis of low molecular weight RNA revealed that transgene-derived small interfering (si) RNA molecules were detected among the poinsettia transformants prior to inoculation. The Agrobacterium-mediated transformation methodology developed in the current study should facilitate improvement of this ornamental plant with enhanced disease resistance, quality improvement and desirable colour alteration. Because poinsettia is a non-food, non-feed plant and is not propagated through sexual reproduction, this is likely to be more acceptable even in areas where genetically modified crops are currently not cultivated. Publication Types: Research Support, Non-U.S. Gov't PMID: 18327592 [PubMed - indexed for MEDLINE] 221: Plant Cell Rep. 2008 Jun;27(6):1039-52. Epub 2008 Mar 8. Morphometric and biochemical characterization of red beet (Beta vulgaris L.) hairy roots obtained after single and double transformations. Thimmaraju R, Venkatachalam L, Bhagyalakshmi N. Plant Cell Biotechnology Department, Central Food Technological Research Institute, Mysore 570 020, India. It is known that T-DNA of Agrobacterium rhizogenes affects processes of plant development and activates the synthesis of secondary metabolites in transformed plant cells. In the present investigation, we provide evidence that different strains of A. rhizogenes significantly affect morphometric, morphological and functional characteristics of hairy roots of red beet (Beta vulgaris L.). Infection with four strains of A. rhizogenes (A4, A 2/83, A 20/83 and LMG-150) resulted in ten clones of hairy roots, which were named accordingly as A4(1), A4(2), A4(3), A 2/83(1), A 2/83(2), A 2/83(3), A 20/83(1), A 20/83(2), A 20/83(3) and LMG-150. Their growth characteristics, pigment content, levels of endogenous auxin and T-DNA copy number showed significant differences probably due to the physiological status of the host cell rather than the T-DNA copy number. Although A 2/83 showed highest hairy root induction capacity, the best hairy root clone was obtained with strain LMG-150 that produced highest biomass and pigments. In this root clone, the enzyme peroxidase was found involved in altering the endogenous auxin pool. When root clone LMG-150 was re-transformed to insert additional individual rol genes, two double transformed clones were obtained, one for rolABC and the other for rolC gene where the former produced higher biomass and betalaine than the latter. Despite the established fact that rol genes of T-DNA influence endogenous phytohormones, no direct correlation among the single transformants and the double transformants was found. This is the first report, in our knowledge, where a hairy root clone has been used to obtain double transformants. Publication Types: Research Support, Non-U.S. Gov't PMID: 18327591 [PubMed - indexed for MEDLINE] 222: Nat Biotechnol. 2008 Mar;26(3):272-4. The 20-year African biotech plan. Singh JA, Daar AS. Publication Types: Letter PMID: 18327231 [PubMed - indexed for MEDLINE] 223: Nat Biotechnol. 2008 Mar;26(3):261-3. Erratum in: Nat Biotechnol. 2008 Apr;26(4):470. Vintage genetic engineering. DeFrancesco L. Publication Types: News PMID: 18327227 [PubMed - indexed for MEDLINE] 224: Nat Biotechnol. 2008 Mar;26(3):259. Erratum in: Nat Biotechnol. 2008 Jun;26(6):709. Tony Conner. Kling J. Publication Types: Biography Historical Article Personal Name as Subject: Connor T PMID: 18327225 [PubMed - indexed for MEDLINE] 225: Nat Biotechnol. 2008 Mar;26(3):255. Plant biotechs defect. Aldridge S. Publication Types: News PMID: 18327220 [PubMed - indexed for MEDLINE] 226: Nat Biotechnol. 2008 Mar;26(3):249-50. Cloned animals deemed safe to eat, but labeling issues loom. Fox JL. Publication Types: News PMID: 18327214 [PubMed - indexed for MEDLINE] 227: Nat Biotechnol. 2008 Mar;26(3):247. Comment in: Nat Biotechnol. 2008 May;26(5):499-500. Nat Biotechnol. 2008 May;26(5):500-1. Nat Biotechnol. 2008 Oct;26(10):1068-70. Off the rails. [No authors listed] Publication Types: Editorial PMID: 18327213 [PubMed - indexed for MEDLINE] 228: J Vet Med Educ. 2007 Winter;34(5):658-73. Integrating molecular biology into the veterinary curriculum. Ryan MT, Sweeney T. College of Life Sciences, School of Agriculture, Food Science and Veterinary Medicine, University College Dublin, Dublin, Ireland. marion.ryan@ucd.ie The modern discipline of molecular biology is gaining increasing relevance in the field of veterinary medicine. This trend must be reflected in the curriculum if veterinarians are to capitalize on opportunities arising from this field and direct its development toward their own goals as a profession. This review outlines current applications of molecular-based technologies that are relevant to the veterinary profession. In addition, the current techniques and technologies employed within the field of molecular biology are discussed. Difficulties associated with teaching a subject such as molecular biology within a veterinary curriculum can be alleviated by effectively integrating molecular topics throughout the curriculum, pitching the subject at an appropriate depth, and employing varied teaching methods throughout. Publication Types: Review PMID: 18326779 [PubMed - indexed for MEDLINE] 229: Food Chem Toxicol. 2008 Jun;46(6):1976-84. Epub 2008 Feb 2. Impact of Bacillus thuringiensis toxin Cry1Ab on rumen epithelial cells (REC) - a new in vitro model for safety assessment of recombinant food compounds. Bondzio A, Stumpff F, Schön J, Martens H, Einspanier R. Department of Veterinary Biochemistry, Freie Universität Berlin, Oertzenweg 19b, 14163 Berlin, Germany. bondzio@zedat.fu-berlin.de The growing use of genetically modified crops necessitates viable screening methods for safety evaluation of recombinant feed, particularly for ruminants. A new sheep rumen epithelial cell culture is introduced as an in vitro cell system for safety evaluation especially focussing on feed and food compounds. We used lactate dehydrogenase (LDH) release, WST-1 conversion, ATP content and caspase 3/7 activity to evaluate cytotoxicity of Cry1Ab, one of the newly expressed Bt-proteins in transgene maize. The results were compared to the effects of valinomycin, a potassium ionophore known to induce cytotoxic effects on a wide range of cells. Whereas no toxicity of Cry1Ab was observed in short as well as in long term experiments, even at non-physiological high concentrations, exposure to valinomycin induced apoptosis and a significant response of all viability parameters after a number of hours. The ATP content and the WST-1 conversion reflecting the energy metabolism of the cells appear to be more sensitive indicators of valinomycin toxicity than the LDH release, a parameter which reflects the membrane integrity. This study presents an in vitro model system, that may be useful as a supplementary tool in toxicity screening before testing substances on animals in vivo. Publication Types: Research Support, Non-U.S. Gov't PMID: 18325653 [PubMed - indexed for MEDLINE] 230: BMC Biotechnol. 2008 Mar 6;8:26. Comparison of different real-time PCR chemistries and their suitability for detection and quantification of genetically modified organisms. Buh Gasparic M, Cankar K, Zel J, Gruden K. Department of Biotechnology and Systems Biology, National Institute of Biology, Vecna pot 111, SI-1000 Ljubljana, Slovenia. meti.buh.gasparic@nib.si BACKGROUND: The real-time polymerase chain reaction is currently the method of choice for quantifying nucleic acids in different DNA based quantification applications. It is widely used also for detecting and quantifying genetically modified components in food and feed, predominantly employing TaqMan and SYBR Green real-time PCR chemistries. In our study four alternative chemistries: Lux, Plexor, Cycling Probe Technology and LNA were extensively evaluated and compared using TaqMan chemistry as a reference system. RESULTS: Amplicons were designed on the maize invertase gene and the 5'-junction of inserted transgene and plant genomic DNA in MON 810 event. Real-time assays were subsequently compared for their efficiency in PCR amplification, limits of detection and quantification, repeatability and accuracy to test the performance of the assays. Additionally, the specificity of established assays was checked on various transgenic and non-transgenic plant species. The overall applicability of the designed assays was evaluated, adding practicability and costs issues to the performance characteristics. CONCLUSION: Although none of the chemistries significantly outperformed the others, there are certain characteristics that suggest that LNA technology is an alternative to TaqMan when designing assays for quantitative analysis. Because LNA probes are much shorter they might be especially appropriate when high specificity is required and where the design of a common TaqMan probe is difficult or even impossible due to sequence characteristics. Plexor on the other hand might be a method of choice for qualitative analysis when sensitivity, low cost and simplicity of use prevail. Publication Types: Comparative Study Evaluation Studies Research Support, Non-U.S. Gov't PMID: 18325084 [PubMed - indexed for MEDLINE] 231: Eur J Nutr. 2008 Mar;47(2):99-103. Epub 2008 Mar 4. Zeaxanthin is bioavailable from genetically modified zeaxanthin-rich potatoes. Bub A, Möseneder J, Wenzel G, Rechkemmer G, Briviba K. Federal Research Centre for Nutrition and Food, Institute of Nutritional Physiology, Haid-und-Neu-Str. 9, 76131 Karlsruhe, Germany. achim.bub@bfel.de The carotenoid zeaxanthin accumulates in the human macula lutea and protects retinal cells from blue light damage. However, zeaxanthin intake from food sources is low. Increasing zeaxanthin in common foods such as potatoes by traditional plant breeding or by genetic engineering could contribute to an increased intake of this carotenoid and, consequently, to a decreased risk of age-related macular degeneration. Our aim was to investigate whether zeaxanthin from genetically modified zeaxanthin-rich potatoes is bioavailable in humans. Three men participated in this randomized, controlled double-blinded, crossover pilot study. All subjects consumed 1,100 g of mashed potatoes, either genetically modified (Solanum tuberosum L. var. Baltica GM47/18; 3 mg zeaxanthin) or wild-type control potatoes (Solanum tuberosum L. var. Baltica; 0.14 mg zeaxanthin). A second treatment was followed after a 7-day wash-out period. The concentration of zeaxanthin was significantly increased in chylomicrons after consumption of genetically modified potatoes and 0.27 mg of the 3 mg zeaxanthin dose could be detected in chylomicrons. Consumption of control potatoes had no effect on concentrations of zeaxanthin in chylomicrons. After normalization of chylomicron zeaxanthin for plasma triacylglycerol, the time course of zeaxanthin concentrations peaked at 7 h after consumption of genetically modified potatoes. There were no significant differences in the concentrations of other major potato carotenoids such as lutein and beta-carotene in chylomicrons after consumption of genetically modified and wild type control potatoes. Thus, consumption of zeaxanthin-rich potatoes significantly increases chylomicron zeaxanthin concentrations suggesting that potentially such potatoes could be used as an important dietary source of zeaxanthin. Publication Types: Randomized Controlled Trial Research Support, Non-U.S. Gov't PMID: 18320254 [PubMed - indexed for MEDLINE] 232: Curr Opin Plant Biol. 2008 Apr;11(2):166-70. Epub 2008 Mar 7. Biofortified crops to alleviate micronutrient malnutrition. Mayer JE, Pfeiffer WH, Beyer P. Center for Applied Biosciences, University of Freiburg, D-79104 Freiburg, Germany. Micronutrient malnutrition affects more than half of the world population, particularly in developing countries. Concerted international and national fortification and supplementation efforts to curb the scourge of micronutrient malnutrition are showing a positive impact, alas without reaching the goals set by international organizations. Biofortification, the delivery of micronutrients via micronutrient-dense crops, offers a cost-effective and sustainable approach, complementing these efforts by reaching rural populations. Bioavailable micronutrients in the edible parts of staple crops at concentrations high enough to impact on human health can be obtained through breeding, provided that sufficient genetic variation for a given trait exists, or through transgenic approaches. Research and breeding programs are underway to enrich the major food staples in developing countries with the most important micronutrients: iron, provitamin A, zinc and folate. Publication Types: Review PMID: 18314378 [PubMed - indexed for MEDLINE] 233: J Immunol Methods. 2008 Apr 20;333(1-2):156-66. Epub 2008 Feb 20. Hybridoma populations enriched for affinity-matured human IgGs yield high-affinity antibodies specific for botulinum neurotoxins. Adekar SP, Jones RM, Elias MD, Al-Saleem FH, Root MJ, Simpson LL, Dessain SK. Cardeza Foundation for Hematologic Research and Kimmel Cancer Center, Thomas Jefferson University, 1015 Walnut Street, Philadelphia, PA 19107, United States. The affinity-matured human antibody repertoire may be ideal as a source for antibody therapeutics against infectious diseases and bioterror agents. Hybridoma methods for cloning these antibodies have many potential advantages, including convenience, high-yield antibody expression, and the ability to capture the antibodies in their native configurations. However, they have been hindered by hybridoma instability and limited accessibility of antigen-specific, class-switched human B-cells. Here, we describe an efficient, three-step method that uses human peripheral blood B-cells to produce stable hybridoma populations that are highly-enriched for affinity-matured human IgG antibodies. Peripheral blood mononuclear cells (PBMCs) are (a) selected for expression of CD27, a marker of post-germinal center B-cells, (b) cultured in vitro to promote B-cell proliferation and class-switching, and (c) fused to a genetically modified myeloma cell line. Using this strategy, we cloned 5 IgG antibodies that bind botulinum neurotoxins (BoNT), the causes of the food-borne paralytic illness, botulism, and Category A Select Bioterror agents. Two of these antibodies bind BoNT with low picomolar affinities. One (30B) is the first high-affinity human antibody to bind serotype B BoNT, and another (6A) is able to neutralize a lethal dose of serotype A BoNT in vivo in pre- and post-exposure models. This optimized hybridoma method will broadly enable access to the native human antibody repertoire. Publication Types: Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't PMID: 18313069 [PubMed - indexed for MEDLINE] 234: Learn Mem. 2008 Feb 28;15(3):133-42. Print 2008 Mar. Visual pattern memory requires foraging function in the central complex of Drosophila. Wang Z, Pan Y, Li W, Jiang H, Chatzimanolis L, Chang J, Gong Z, Liu L. State Key Laboratory of Brain and Cognitive Science, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China. The role of the foraging (for) gene, which encodes a cyclic guanosine-3',5'-monophosphate (cGMP)-dependent protein kinase (PKG), in food-search behavior in Drosophila has been intensively studied. However, its functions in other complex behaviors have not been well-characterized. Here, we show experimentally in Drosophila that the for gene is required in the operant visual learning paradigm. Visual pattern memory was normal in a natural variant rover (for(R)) but was impaired in another natural variant sitter (for(S)), which has a lower PKG level. Memory defects in for(S) flies could be rescued by either constitutive or adult-limited expression of for in the fan-shaped body. Interestingly, we showed that such rescue also occurred when for was expressed in the ellipsoid body. Additionally, expression of for in the fifth layer of the fan-shaped body restored sufficient memory for the pattern parameter "elevation" but not for "contour orientation," whereas expression of for in the ellipsoid body restored sufficient memory for both parameters. Our study defines a Drosophila model for further understanding the role of cGMP-PKG signaling in associative learning/memory and the neural circuit underlying this for-dependent visual pattern memory. Publication Types: Research Support, Non-U.S. Gov't PMID: 18310460 [PubMed - indexed for MEDLINE] 235: J Agric Food Chem. 2008 Mar 26;56(6):1818-28. Epub 2008 Feb 28. Development of a real-time PCR method for the differential detection and quantification of four solanaceae in GMO analysis: potato (Solanum tuberosum), tomato (Solanum lycopersicum), eggplant (Solanum melongena), and pepper (Capsicum annuum). Chaouachi M, El Malki R, Berard A, Romaniuk M, Laval V, Brunel D, Bertheau Y. Unité Etude du Polymorphisme des Génomes Végétaux (EPGV) UR1279, Centre National de Génotypage (CNG), 2 rue Gaston Crémieux 91057, CP5721, Evry cedex, France. The labeling of products containing genetically modified organisms (GMO) is linked to their quantification since a threshold for the presence of fortuitous GMOs in food has been established. This threshold is calculated from a combination of two absolute quantification values: one for the specific GMO target and the second for an endogenous reference gene specific to the taxon. Thus, the development of reliable methods to quantify GMOs using endogenous reference genes in complex matrixes such as food and feed is needed. Plant identification can be difficult in the case of closely related taxa, which moreover are subject to introgression events. Based on the homology of beta-fructosidase sequences obtained from public databases, two couples of consensus primers were designed for the detection, quantification, and differentiation of four Solanaceae: potato (Solanum tuberosum), tomato (Solanum lycopersicum), pepper (Capsicum annuum), and eggplant (Solanum melongena). Sequence variability was studied first using lines and cultivars (intraspecies sequence variability), then using taxa involved in gene introgressions, and finally, using taxonomically close taxa (interspecies sequence variability). This study allowed us to design four highly specific TaqMan-MGB probes. A duplex real time PCR assay was developed for simultaneous quantification of tomato and potato. For eggplant and pepper, only simplex real time PCR tests were developed. The results demonstrated the high specificity and sensitivity of the assays. We therefore conclude that beta-fructosidase can be used as an endogenous reference gene for GMO analysis. Publication Types: Comparative Study Research Support, Non-U.S. Gov't PMID: 18303841 [PubMed - indexed for MEDLINE] 236: Proc Natl Acad Sci U S A. 2008 Mar 4;105(9):3640-5. Epub 2008 Feb 26. Microarray analyses reveal that plant mutagenesis may induce more transcriptomic changes than transgene insertion. Batista R, Saibo N, Lourenço T, Oliveira MM. Instituto Nacional de Saúde Dr. Ricardo Jorge, Avenida Padre Cruz, 1649-016 Lisbon, Portugal. rita.batista@insa.min-saude.pt Controversy regarding genetically modified (GM) plants and their potential impact on human health contrasts with the tacit acceptance of other plants that were also modified, but not considered as GM products (e.g., varieties raised through conventional breeding such as mutagenesis). What is beyond the phenotype of these improved plants? Should mutagenized plants be treated differently from transgenics? We have evaluated the extent of transcriptome modification occurring during rice improvement through transgenesis versus mutation breeding. We used oligonucleotide microarrays to analyze gene expression in four different pools of four types of rice plants and respective controls: (i) a gamma-irradiated stable mutant, (ii) the M1 generation of a 100-Gy gamma-irradiated plant, (iii) a stable transgenic plant obtained for production of an anticancer antibody, and (iv) the T1 generation of a transgenic plant produced aiming for abiotic stress improvement, and all of the unmodified original genotypes as controls. We found that the improvement of a plant variety through the acquisition of a new desired trait, using either mutagenesis or transgenesis, may cause stress and thus lead to an altered expression of untargeted genes. In all of the cases studied, the observed alteration was more extensive in mutagenized than in transgenic plants. We propose that the safety assessment of improved plant varieties should be carried out on a case-by-case basis and not simply restricted to foods obtained through genetic engineering. Publication Types: Comparative Study PMID: 18303117 [PubMed - indexed for MEDLINE] 237: Adv Biochem Eng Biotechnol. 2008;111:149-86. Plant biotechnology: transgenic crops. Shewry PR, Jones HD, Halford NG. Rothamsted Research, AL5 2JQ, Harpenden, Herts, UK. peter.shewry@bbsrc.ac.uk Transgenesis is an important adjunct to classical plant breeding, in that it allows the targeted manipulation of specific characters using genes from a range of sources. The current status of crop transformation is reviewed, including methods of gene transfer, the selection of transformed plants and control of transgene expression. The application of genetic modification technology to specific traits is then discussed, including input traits relating to crop production (herbicide tolerance and resistance to insects, pathogens and abiotic stresses) and output traits relating to the composition and quality of the harvested organs. The latter include improving the nutritional quality for consumers as well as the improvement of functional properties for food processing. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 18299808 [PubMed - indexed for MEDLINE] 238: Plant Biotechnol J. 2008 May;6(4):337-45. Epub 2008 Feb 19. The use of life-cycle assessment to evaluate the environmental impacts of growing genetically modified, nitrogen use-efficient canola. Strange A, Park J, Bennett R, Phipps R. School of Agriculture, Policy and Development, The University of Reading, PO Box 237, Reading RG6 6AR, UK. Agriculture, particularly intensive crop production, makes a significant contribution to environmental pollution. A variety of canola (Brassica napus) has been genetically modified to enhance nitrogen use efficiency, effectively reducing the amount of fertilizer required for crop production. A partial life-cycle assessment adapted to crop production was used to assess the potential environmental impacts of growing genetically modified, nitrogen use-efficient (GMNUE) canola in North Dakota and Minnesota compared with a conventionally bred control variety. The analysis took into account the entire production system used to produce 1 tonne of canola. This comprised raw material extraction, processing and transportation, as well as all agricultural field operations. All emissions associated with the production of 1 tonne of canola were listed, aggregated and weighted in order to calculate the level of environmental impact. The findings show that there are a range of potential environmental benefits associated with growing GMNUE canola. These include reduced impacts on global warming, freshwater ecotoxicity, eutrophication and acidification. Given the large areas of canola grown in North America and, in particular, Canada, as well as the wide acceptance of genetically modified varieties in this area, there is the potential for GMNUE canola to reduce pollution from agriculture, with the largest reductions predicted to be in greenhouse gases and diffuse water pollution. Publication Types: Evaluation Studies Research Support, Non-U.S. Gov't PMID: 18298428 [PubMed - indexed for MEDLINE] 239: Crit Rev Immunol. 2008;28(1):15-43. Scratching the surface: towards understanding the pathogenesis of atopic dermatitis. Sehra S, Tuana FM, Holbreich M, Mousdicas N, Tepper RS, Chang CH, Travers JB, Kaplan MH. Department of Pediatrics and HB Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, IN 46202, USA. Atopic dermatitis (AD) is a chronic inflammatory skin disease with a steadily increasing prevalence affecting 10%-20% of infants and 1%-3% of adults globally. It is often the first clinical manifestation of atopic disease preceding asthma and allergic rhinitis. At least half of the children with AD develop some other form of atopic disease later in life. The pathogenesis of AD involves a complex interplay of factors, including genetic predisposition due to altered immune or skin barrier function, interactions with the environment, and infectious triggers of inflammation. In this review, we summarize the recent advances in understanding the contribution of different factors in the pathophysiology of AD in human and animal model systems. These insights provide new therapeutic potential for the treatment of human AD. Publication Types: Review PMID: 18298382 [PubMed - indexed for MEDLINE] 240: J Agric Food Chem. 2008 Mar 26;56(6):1804-9. Epub 2008 Feb 26. Qualitative and quantitative event-specific PCR detection methods for oxy-235 canola based on the 3' integration flanking sequence. Yang L, Guo J, Zhang H, Liu J, Zhang D. GMO Detection Laboratory, SJTU-Bor Luh Food Safety Center, Ministry of Education, School of Life Science and Biotechnology, Shanghai Jiao Tong University, Shanghai, People's Republic of China. As more genetically modified plant events are approved for commercialization worldwide, the event-specific PCR method has become the key method for genetically modified organism (GMO) identification and quantification. This study reveals the 3' flanking sequence of the exogenous integration of Oxy-235 canola employing thermal asymmetric interlaced PCR (TAIL-PCR). On the basis of the revealed 3' flanking sequence, PCR primers and TaqMan probe were designed and qualitative and quantitative PCR assays were established for Oxy-235 canola. The specificity and limits of detection (LOD) and quantification (LOQ) of these two PCR assays were validated to as low as 0.1% for the relative LOD of qualitative PCR assay; the absolute LOD and LOQ were low to 10 and 20 copies of canola genomic DNA in quantitative PCR assay, respectively. Furthermore, ideal quantified results were obtained in the practical canola sample detection. All of the results indicate that the developed qualitative and quantitative PCR methods based on the revealed 3' integration flanking sequence are suitable for GM canola Oxy-235 identification and quantification. Publication Types: Research Support, Non-U.S. Gov't PMID: 18298073 [PubMed - indexed for MEDLINE] 241: J Agric Food Chem. 2008 Mar 26;56(6):1977-83. Epub 2008 Feb 26. Individual detection of genetically modified maize varieties in non-identity-preserved maize samples. Akiyama H, Sakata K, Kondo K, Tanaka A, Liu MS, Oguchi T, Furui S, Kitta K, Hino A, Teshima R. National Institute of Health Sciences, Tokyo, Japan. akiyama@nihs.go.jp In many countries, the labeling of grains and feed- and foodstuffs is mandatory if the genetically modified organism (GMO) content exceeds a certain level of approved GM varieties. The GMO content in a maize sample containing the combined-trait (stacked) GM maize as determined by the currently available methodology is likely to be overestimated. However, there has been little information in the literature on the mixing level and varieties of stacked GM maize in real sample grains. For the first time, the GMO content of non-identity-preserved (non-IP) maize samples imported from the United States has been successfully determined by using a previously developed individual kernel detection system coupled to a multiplex qualitative PCR method followed by multichannel capillary gel electrophoresis system analysis. To clarify the GMO content in the maize samples imported from the United States, determine how many stacked GM traits are contained therein, and which GM trait varieties frequently appeared in 2005, the GMO content (percent) on a kernel basis and the varieties of the GM kernels in the non-IP maize samples imported from the United States were investigated using the individual kernel analysis system. The average (+/-standard deviation) of the GMO contents on a kernel basis in five non-IP sample lots was determined to be 51.0+/-21.6%, the percentage of a single GM trait grains was 39%, and the percentage of the stacked GM trait grains was 12%. The MON810 grains and NK603 grains were the most frequent varieties in the single GM traits. The most frequent stacked GM traits were the MON810xNK603 grains. In addition, the present study would provide the answer and impact for the quantification of GM maize content in the GM maize kernels on labeling regulation. Publication Types: Research Support, Non-U.S. Gov't PMID: 18298063 [PubMed - indexed for MEDLINE] 242: Soc Sci Med. 2008 Apr;66(8):1797-808. Epub 2008 Feb 21. Potential impacts of iron biofortification in India. Stein AJ, Meenakshi JV, Qaim M, Nestel P, Sachdev HP, Bhutta ZA. University of Hohenheim, Agricultural Economics and Social Sciences, Inst. 490b, 70593 Stuttgart, Germany. academic@ajstein.de Iron deficiency is a widespread nutrition and health problem in developing countries, causing impairments in physical activity and cognitive development, as well as maternal mortality. Although food fortification and supplementation programmes have been effective in some countries, their overall success remains limited. Biofortification, that is, breeding food crops for higher micronutrient content, is a relatively new approach, which has been gaining international attention recently. We propose a methodology for ex ante impact assessment of iron biofortification, building on a disability-adjusted life years (DALYs) framework. This methodology is applied in an Indian context. Using a large and representative data set of household food consumption, the likely effects of iron-rich rice and wheat varieties are simulated for different target groups and regions. These varieties, which are being developed by an international public research consortium, based on conventional breeding techniques, might be ready for local distribution within the next couple of years. The results indicate sizeable potential health benefits. Depending on the underlying assumptions, the disease burden associated with iron deficiency could be reduced by 19-58%. Due to the relatively low institutional cost to reach the target population, the expected cost-effectiveness of iron biofortification compares favourably with other micronutrient interventions. Nonetheless, biofortification should not be seen as a substitute for other interventions. Each approach has its particular strengths, so they complement one another. Publication Types: Research Support, Non-U.S. Gov't PMID: 18291567 [PubMed - indexed for MEDLINE] 243: Rev Panam Salud Publica. 2008 Jan;23(1):52-8. [Academic production on food labeling in Brazil] [Article in Portuguese] Câmara MC, Marinho CL, Guilam MC, Braga AM. Fundação Oswaldo Cruz (Fiocruz), Centro de Estudos da Saúde do Trabalhador e Ecologia Humana (CESTEH), Brazil. maria.clara@ensp.fiocruz.br OBJECTIVE: To review and discuss academic production (theses and dissertations) on the topic of labeling of prepackaged foods in Brazil. METHOD: A search of the database maintained by the Coordination for the Development of Higher Education Professionals (CAPES), one of the two Brazilian government research funding and support agencies, was conducted on the following keywords: "rotulagem" (labeling), "rotulagem nutricional" (food labeling) and "rótulo de alimentos" (food labels). The search covered the years 1987 (earliest year available) to 2004. RESULTS: We identified 49 studies on this topic. Content analysis identified three major themes: the extent to which food labels meet specific legal requirements (57.2%); the degree to which consumers understand the information on labels (22.4%); and the labeling of transgenic or genetically-modified foods (20.4%). CONCLUSIONS: Food labeling is a frequent topic and is adequately covered by the Brazilian academic production. In most of the studies, ineffective law enforcement appears to be the main factor in the lack of compliance with and disrespect for the food labeling rules and regulations in Brazil. Publication Types: English Abstract Review PMID: 18291073 [PubMed - indexed for MEDLINE] 244: Food Chem Toxicol. 2008 May;46(5):1414-36. Epub 2008 Jan 12. Edible safety requirements and assessment standards for agricultural genetically modified organisms. Deng P, Zhou X, Zhou P, Du Z, Hou H, Yang D, Tan J, Wu X, Zhang J, Yang Y, Liu J, Liu G, Li Y, Liu J, Yu L, Fang S, Yang X. Shenzhen Center for Disease Control and Prevention, Shenzhen, China. szdpj2002@163.com This paper describes the background, principles, concepts and methods of framing the technical regulation for edible safety requirement and assessment of agricultural genetically modified organisms (agri-GMOs) for Shenzhen Special Economic Zone in the People's Republic of China. It provides a set of systematic criteria for edible safety requirements and the assessment process for agri-GMOs. First, focusing on the degree of risk and impact of different agri-GMOs, we developed hazard grades for toxicity, allergenicity, anti-nutrition effects, and unintended effects and standards for the impact type of genetic manipulation. Second, for assessing edible safety, we developed indexes and standards for different hazard grades of recipient organisms, for the influence of types of genetic manipulation and hazard grades of agri-GMOs. To evaluate the applicability of these criteria and their congruency with other safety assessment systems for GMOs applied by related organizations all over the world, we selected some agri-GMOs (soybean, maize, potato, capsicum and yeast) as cases to put through our new assessment system, and compared our results with the previous assessments. It turned out that the result of each of the cases was congruent with the original assessment. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 18289760 [PubMed - indexed for MEDLINE] 245: Environ Entomol. 2007 Oct;36(5):1269-74. Response of ground beetle (Coleoptera: Carabidae) field populations to four years of Lepidoptera-specific Bt corn production. Floate KD, Cárcamo HA, Blackshaw RE, Postman B, Bourassa S. Agriculture and Agri-Food Canada, Lethbridge Research Centre, 5403 1st Ave. S., Lethbridge, AB, Canada T1J 4B1. floatek@agr.gc.ca Pitfall traps were used to monitor populations of ground beetles (Coleoptera: Carabidae) in plots of corn grown in continuous cultivation during a 4-yr period (2000-2003). Treatments included transgenic corn expressing a Bt Cry protein with efficacy specific against Lepidoptera (Bt), conventional corn grown with insecticide application (I), and the same conventional cultivar grown without insecticide application (NI). Mixed-model analyses of variance were performed on pitfall captures of beetles combined across weeks to give seasonal sums. Effects of corn treatment were not detected (P > 0.05) on total beetle abundance or species richness in any year. Effects of corn treatment on individual taxa were detected (P < 0.05) for 3 of the 39 species-by-year combinations examined. Effects of near significance (P < 0.08) were detected for an additional two species. In 2001, captures of Amara farcta Leconte and Harpalus amputatus Say were lower in Bt plots than in I or NI plots. In 2003, captures of Amara apricaria (Paykull) and Amara carinata (Leconte) were higher in Bt plots than in I or NI plots. Also in 2003, captures of Poecilus scitulus Leconte were higher in I plots than in Bt or NI plots. These patterns were not repeated among years. Results of this study indicate that cultivation of Lepidoptera-specific Bt corn in southern Alberta does not appreciably affect ground beetle populations. Publication Types: Research Support, Non-U.S. Gov't PMID: 18284752 [PubMed - indexed for MEDLINE] 246: Annu Rev Plant Biol. 2008;59:771-812. Genetically Engineered Plants and Foods: A Scientist's Analysis of the Issues (Part I). Lemaux PG. Department of Plant and Microbial Biology, University of California, Berkeley, California 94720, USA. lemauxpg@nature.berkeley.edu Through the use of the new tools of genetic engineering, genes can be introduced into the same plant or animal species or into plants or animals that are not sexually compatible-the latter is a distinction with classical breeding. This technology has led to the commercial production of genetically engineered (GE) crops on approximately 250 million acres worldwide. These crops generally are herbicide and pest tolerant, but other GE crops in the pipeline focus on other traits. For some farmers and consumers, planting and eating foods from these crops are acceptable; for others they raise issues related to safety of the foods and the environment. In Part I of this review some general and food issues raised regarding GE crops and foods will be addressed. Responses to these issues, where possible, cite peer-reviewed scientific literature. In Part II to appear in 2009, issues related to environmental and socioeconomic aspects of GE crops and foods will be covered. Publication Types: Review PMID: 18284373 [PubMed - indexed for MEDLINE] 247: PLoS Genet. 2008 Feb;4(2):e24. A role for autophagy in the extension of lifespan by dietary restriction in C. elegans. Hansen M, Chandra A, Mitic LL, Onken B, Driscoll M, Kenyon C. Department of Biochemistry and Biophysics, University of California San Francisco, San Francisco, California, United States of America. In many organisms, dietary restriction appears to extend lifespan, at least in part, by down-regulating the nutrient-sensor TOR (Target Of Rapamycin). TOR inhibition elicits autophagy, the large-scale recycling of cytoplasmic macromolecules and organelles. In this study, we asked whether autophagy might contribute to the lifespan extension induced by dietary restriction in C. elegans. We find that dietary restriction and TOR inhibition produce an autophagic phenotype and that inhibiting genes required for autophagy prevents dietary restriction and TOR inhibition from extending lifespan. The longevity response to dietary restriction in C. elegans requires the PHA-4 transcription factor. We find that the autophagic response to dietary restriction also requires PHA-4 activity, indicating that autophagy is a transcriptionally regulated response to food limitation. In spite of the rejuvenating effect that autophagy is predicted to have on cells, our findings suggest that autophagy is not sufficient to extend lifespan. Long-lived daf-2 insulin/IGF-1 receptor mutants require both autophagy and the transcription factor DAF-16/FOXO for their longevity, but we find that autophagy takes place in the absence of DAF-16. Perhaps autophagy is not sufficient for lifespan extension because although it provides raw material for new macromolecular synthesis, DAF-16/FOXO must program the cells to recycle this raw material into cell-protective longevity proteins. Publication Types: Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't PMID: 18282106 [PubMed - indexed for MEDLINE] 248: Nat Protoc. 2008;3(2):181-9. Agrobacterium-mediated transformation of Brassica napus and Brassica oleracea. Bhalla PL, Singh MB. Australian Research Council Centre of Excellence for Integrative Legume Research, Faculty of Land and Food Resources, The University of Melbourne, Parkville, Victoria 3010, Australia. premlb@unimelb.edu.au Agrobacterium-mediated transformation is widely used for gene delivery in plants. However, commercial cultivars of crop plants are often recalcitrant to transformation because the protocols established for model varieties are not directly applicable to them. The genus Brassica includes the oil seed crop, canola (B. napus), and vegetable crop varieties of Brassica oleracea, including cauliflower, broccoli and cabbage. Here, we describe an efficient protocol for Agrobacterium-mediated transformation using seedling explants that is applicable to various Brassica varieties; this protocol has been used to genetically engineer commercial cultivars of canola and cauliflower in our laboratory. Young seedling explants are inoculated with Agrobacterium on the day of explant preparation. Explants are grown for 1 week in the absence of a selective agent before being transferred to a selective medium to recover transgenic shoots. Transgenic shoots are subjected to an additional round of selection on medium containing higher levels of the selective agent and a low-carbohydrate source; this helps to eliminate false-positive plants. Use of seedling explants offers flexible experiment planning and a convenient explant source. Using this protocol, transgenic plants can be obtained in 2.5 to 3.5 months. Publication Types: Research Support, Non-U.S. Gov't PMID: 18274519 [PubMed - indexed for MEDLINE] 249: Foreign Aff. 2000;79(3):24-38. The global food fight. Paarlberg R. Publication Types: Historical Article PMID: 18274013 [PubMed - indexed for MEDLINE] 250: Pest Manag Sci. 2008 Apr;64(4):360-5. Evolved glyphosate-resistant weeds around the world: lessons to be learnt. Powles SB. WA Herbicide Resistance Initiative, School of Plant Biology, University of Western Australia, Perth 6014, Australia. spowles@plants.uwa.edu.au Glyphosate is the world's most important herbicide, with many uses that deliver effective and sustained control of a wide spectrum of unwanted (weedy) plant species. Until recently there were relatively few reports of weedy plant species evolving resistance to glyphosate. Since 1996, the advent and subsequent high adoption of transgenic glyphosate-resistant crops in the Americas has meant unprecedented and often exclusive use of glyphosate for weed control over very large areas. Consequently, in regions of the USA where transgenic glyphosate-resistant crops dominate, there are now evolved glyphosate-resistant populations of the economically damaging weed species Ambrosia artemissifolia L., Ambrosia trifida L., Amaranthus palmeri S Watson, Amaranthus rudis JD Sauer, Amaranthus tuberculatus (Moq) JD Sauer and various Conyza and Lolium spp. Likewise, in areas of transgenic glyphosate-resistant crops in Argentina and Brazil, there are now evolved glyphosate-resistant populations of Sorghum halepense (L.) Pers and Euphorbia heterophylla L. respectively. As transgenic glyphosate-resistant crops will remain very popular with producers, it is anticipated that glyphosate-resistant biotypes of other prominent weed species will evolve over the next few years. Therefore, evolved glyphosate-resistant weeds are a major risk for the continued success of glyphosate and transgenic glyphosate-resistant crops. However, glyphosate-resistant weeds are not yet a problem in many parts of the world, and lessons can be learnt and actions taken to achieve glyphosate sustainability. A major lesson is that maintenance of diversity in weed management systems is crucial for glyphosate to be sustainable. Glyphosate is essential for present and future world food production, and action to secure its sustainability for future generations is a global imperative. Copyright (c) 2008 Society of Chemical Industry. Publication Types: Review PMID: 18273881 [PubMed - indexed for MEDLINE] 251: Mycopathologia. 2008 Feb;165(2):89-97. Epub 2008 Feb 12. Development of a GFP-expressing Aspergillus flavus strain to study fungal invasion, colonization, and resistance in cottonseed. Rajasekaran K, Cary JW, Cotty PJ, Cleveland TE. USDA, ARS, Southern Regional Research Center, Food and Feed Safety Research Unit, 1100 Robert E. Lee Blvd., New Orleans, LA 70124, USA. krajah@srrc.ars.usda.gov Cotton bolls were inoculated with a green fluorescent protein (GFP)-expressing Aspergillus flavus (strain 70) to monitor fungal growth, mode of entry, colonization of cottonseeds, and production of aflatoxins. The GFP strain and the wild-type did not differ significantly in pathogen aggressiveness as indicated by similar reductions in inoculated locule weight. GFP fluorescence was at least 10 times higher than the blue green yellow fluorescence (BGYF) produced in response to infection by A. flavus. The GFP produced by the strain made it possible to identify and monitor specific plant tissues colonized by the fungus. For example, the inner seed coat and cotyledon were colonized by the fungus within 72 h of inoculation and the mode of entry was invariably through the porous chalazal cap in intact seeds. The amount of GFP fluorescence was shown to be an indicator of fungal growth, colonization and, to some extent, aflatoxin production. The A. flavus strain expressing GFP should be very useful for rapidly identifying cotton lines with enhanced resistance to A. flavus colonization developed through genetic engineering or traditional plant breeding. In addition, development of GFP expressing A. flavus strain provides an easy and rapid assay procedure for studying the ecology, etiology, and epidemiology of cotton boll rot caused by A. flavus resulting in aflatoxin contamination. PMID: 18266076 [PubMed - indexed for MEDLINE] 252: Nat Biotechnol. 2008 Feb;26(2):161-2. Comment in: Nat Biotechnol. 2008 Aug;26(8):858-9. Action needed to harmonize regulation of low-level presence of biotech traits. Krueger R, Le Buanec B. Publication Types: Letter PMID: 18259165 [PubMed - indexed for MEDLINE] 253: Nat Biotechnol. 2008 Feb;26(2):159-60. FDA on transgenic animals--a dog's breakfast? Miller HI. Publication Types: Letter PMID: 18259164 [PubMed - indexed for MEDLINE] 254: Nat Biotechnol. 2008 Feb;26(2):139-40. Scientists cry foul as Europe plays politics with GM crops. Hodgson J. Publication Types: News PMID: 18259156 [PubMed - indexed for MEDLINE] 255: Food Chem Toxicol. 2007 Dec 25. [Epub ahead of print] Identification of potentially emerging food safety issues by analysis of reports published by the European Community's Rapid Alert System for Food and Feed (RASFF) during a four-year period. Kleter GA, Prandini A, Filippi L, Marvin HJ. RIKILT – Institute of Food Safety, Wageningen University and Research Center, P.O. Box 230, NL-6700 AE Wageningen, The Netherlands. The SAFE FOODS project undertakes to design a new approach towards the early identification of emerging food safety hazards. This study explored the utility of notifications filed through RASFF, the European Commission's Rapid Alert System for Food and Feed, to identify emerging trends in food safety issues. RASFF information and alert notifications published in the four-year period of July 2003-June 2007 were assigned to categories of products and hazards. For chronological trend analysis, a basic time unit of three months was chosen. Data within each hazard category were analyzed for chronological trends, relationships between product and hazard categories, regions of origin, and countries filing the notifications. Conspicuous trends that were observed included a rise in the incidence of food contact substances, particularly 2-isopropyl-thioxanthone, as well as of chemical substances migrating from utensils and fraud-related issues. Temporary increases were noted in the incidences of the unauthorized dye Para Red, genetically modified organisms, the pesticide isophenfos-methyl, and herring worm, Anisakis simplex. National and European authorities themselves have signaled these conspicuous trends and taken measures. It is recommended to add complementary data to RASFF data, including safety assessments, risk management measures, background data on hazards and surveillance patterns, for a holistic approach towards early identification of emerging hazards. PMID: 18255210 [PubMed - as supplied by publisher] 256: Physiol Plant. 2008 Feb;132(2):236-53. Transcriptome analyses give insights into selenium-stress responses and selenium tolerance mechanisms in Arabidopsis. Van Hoewyk D, Takahashi H, Inoue E, Hess A, Tamaoki M, Pilon-Smits EA. Department of Biology, Colorado State University, Fort Collins, CO 80523, USA. Selenate is chemically similar to sulfate and can be taken up and assimilated by plants via the same transporters and enzymes. In contrast to many other organisms, selenium (Se) has not been shown to be essential for higher plants. In excess, Se is toxic and restricts development. Both Se deficiency and toxicity pose problems worldwide. To obtain better insights into the effects of Se on plant metabolism and into plant mechanisms involved in Se tolerance, the transcriptome of Arabidopsis plants grown with or without selenate was studied and Se-responsive genes identified. Roots and shoots exhibited different Se-related changes in gene regulation and metabolism. Many genes involved in sulfur (S) uptake and assimilation were upregulated. Accordingly, Se treatment enhanced sulfate levels in plants, but the quantity of organic S metabolites decreased. Transcripts regulating the synthesis and signaling of ethylene and jasmonic acid were also upregulated by Se. Arabidopsis mutants defective in ethylene or jasmonate response pathways exhibited reduced tolerance to Se, suggesting an important role for these two stress hormones in Se tolerance. Selenate upregulated a variety of transcripts that were also reportedly induced by salt and osmotic stress. Selenate appeared to repress plant development, as suggested by the downregulation of genes involved in cell wall synthesis and auxin-regulated proteins. The Se-responsive genes discovered in this study may help create plants that can better tolerate and accumulate Se, which may enhance the effectiveness of Se phytoremediation or serve as Se-fortified food. Publication Types: Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. PMID: 18251864 [PubMed - indexed for MEDLINE] 257: Appl Environ Microbiol. 2008 Apr;74(7):2129-34. Epub 2008 Feb 1. Construction of sterile ime1Delta-transgenic Saccharomyces cerevisiae wine yeasts unable to disseminate in nature. Ramírez M, Ambrona J. Departamento de Microbiología (Antiguo Rectorado), Facultad de Ciencias, Universidad de Extremadura, 06071 Badajoz, Spain. mramirez@unex.es The use of new transgenic yeasts in industry carries a potential environmental risk because their dispersal, introducing new artificial genetic combinations into nature, could have unpredictable consequences. This risk could be avoided by using sterile transgenic yeasts that are unable to sporulate and mate with wild yeasts. These sterile yeasts would not survive the annual cyclic harvesting periods, being condemned to disappear in the wineries and vineyards in less than a year. We have constructed new ime1Delta wine yeasts that are unable to sporulate and mate, bear easy-to-detect genetic markers, and quickly disappear in grape must fermentation immediately after sporulation of the yeast population. These sterile yeasts maintained the same biotechnological properties as their parent yeasts without any detectable deleterious effect of the ime1Delta mutation. These yeasts are therefore interesting biotechnologically for food industry applications and for genetically modified microorganism environmental monitoring studies. Publication Types: Research Support, Non-U.S. Gov't PMID: 18245242 [PubMed - indexed for MEDLINE] 258: Appetite. 2008 Jul;51(1):58-68. Epub 2007 Dec 15. Knowledge, attitudes towards and acceptability of genetic modification in Germany. Christoph IB, Bruhn M, Roosen J. Federal Research Centre for Nutrition and Food (BfEL), Department of Food Economics, Hermann-Weigmann-Strasse, 24103 Kiel, Germany. Genetic modification remains a controversial issue. The aim of this study is to analyse the attitudes towards genetic modification, the knowledge about it and its acceptability in different application areas among German consumers. Results are based on a survey from spring 2005. An exploratory factor analysis is conducted to identify the attitudes towards genetic modification. The identified factors are used in a cluster analysis that identified a cluster of supporters, of opponents and a group of indifferent consumers. Respondents' knowledge of genetics and biotechnology differs among the found clusters without revealing a clear relationship between knowledge and support of genetic modification. The acceptability of genetic modification varies by application area and cluster, and genetically modified non-food products are more widely accepted than food products. The perception of personal health risks has high explanatory power for attitudes and acceptability. Publication Types: Research Support, Non-U.S. Gov't PMID: 18243411 [PubMed - indexed for MEDLINE] 259: Trends Biotechnol. 2008 Mar;26(3):122-5. Is biotechnology a victim of anti-science bias in scientific journals? Miller HI, Morandini P, Ammann K. The Hoover Institution, 434 Galvez Mall, Stanford University, Stanford, CA 94305-6010, USA. miller@hoover.stanford.edu Primarily outside the scientific community, misapprehensions and misinformation about recombinant DNA-modified (also known as 'genetically modified', or 'GM') plants have generated significant 'pseudo-controversy' over their safety that has resulted in unscientific and excessive regulation (with attendant inflated development costs) and disappointing progress. But pseudo-controversy and sensational claims have originated within the scientific community as well, and even scholarly journals' treatment of the subject has been at times unscientific, one-sided and irresponsible. These shortcomings have helped to perpetuate 'The Big Lie' - that recombinant DNA technology applied to agriculture and food production is unproven, unsafe, untested, unregulated and unwanted. Those misconceptions, in turn, have given rise to unwarranted opposition and tortuous, distorted public policy. Publication Types: Review PMID: 18243381 [PubMed - indexed for MEDLINE] 260: Anal Bioanal Chem. 2008 Oct;392(3):347-54. Epub 2008 Feb 2. Advances in molecular techniques for the detection and quantification of genetically modified organisms. Elenis DS, Kalogianni DP, Glynou K, Ioannou PC, Christopoulos TK. Department of Chemistry, University of Athens, Athens, 15771, Greece. Progress in genetic engineering has led to the introduction of genetically modified organisms (GMOs) whose genomes have been altered by the integration of a novel sequence conferring a new trait. To allow consumers an informed choice, many countries require food products to be labeled if the GMO content exceeds a certain threshold. Consequently, the development of analytical methods for GMO screening and quantification is of great interest. Exponential amplification by the polymerase chain reaction (PCR) remains a central step in molecular methods of GMO detection and quantification. In order to meet the challenge posed by the continuously increasing number of GMOs, various multiplex assays have been developed for the simultaneous amplification and/or detection of several GMOs. Classical agarose gel electrophoresis is being replaced by capillary electrophoresis (CE) systems, including CE chips, for the rapid and automatable separation of amplified fragments. Microtiter well-based hybridization assays allow high-throughput analysis of many samples in a single plate. Microarrays have been introduced in GMO screening as a technique for the simultaneous multianalyte detection of amplified sequences. Various types of biosensors, including surface plasmon resonance sensors, quartz crystal microbalance piezoelectric sensors, thin-film optical sensors, dry-reagent dipstick-type sensors and electrochemical sensors were introduced in GMO screening because they offer simplicity and lower cost. GMO quantification is performed by real-time PCR (rt-QPCR) and competitive PCR. New endogenous reference genes have been validated. rt-QPCR is the most widely used approach. Multiplexing is another trend in this field. Strategies for high-throughput multiplex competitive quantitative PCR have been reported. Publication Types: Review PMID: 18239909 [PubMed - indexed for MEDLINE] 261: Peptides. 2008 Mar;29(3):331-7. Epub 2007 Dec 3. Anti-hypertensive activity of genetically modified soybean seeds accumulating novokinin. Yamada Y, Nishizawa K, Yokoo M, Zhao H, Onishi K, Teraishi M, Utsumi S, Ishimoto M, Yoshikawa M. Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Uji, Kyoto, Japan. yyamada@kais.kyoto-u.ac.jp Novokinin (Arg-Pro-Leu-Lys-Pro-Trp), which has been designed based on the structure of ovokinin (2-7), significantly reduces the systolic blood pressure at a dose of 100 microg/kg after oral administration in spontaneously hypertensive rats (SHRs). In this study, we generated a transgenic soybean which accumulates novokinin. A vector encoding a modified beta-conglycinin alpha' subunit (4novokinin-alpha') in which four novokinin sequences have been incorporated by site-directed mutagenesis was introduced into somatic embryos by whisker-mediated gene transformation to produce a transgenic soybean. The 4novokinin-alpha' occupied 0.5% of total soluble protein and 5% of the beta-conglycinin alpha' subunit in the transgenic soybean seeds. Protein extracted from the transgenic soybean reduced systolic blood pressure after single oral administration in SHRs at a dose of 0.15 g/kg. Defatted flour from the transgenic soybean also reduced the systolic blood pressure at a dose of 0.25 g/kg. Thus, the 4novokinin-alpha' produced in soybean exhibited an anti-hypertensive activity in SHRs after oral administration. Publication Types: Research Support, Non-U.S. Gov't PMID: 18226422 [PubMed - indexed for MEDLINE] 262: Bull Acad Natl Med. 2007 Apr-May;191(4-5):807-14; discussion 814. [Allergic risk and role of the Allergy Vigilance Network] [Article in French] Moneret-Vautrin DA. Médecine interne, immunologie clinique et allergologique, Hôpital Central, 54035 Nancy cedex. The recent increase in the incidence of severe anaphylaxis calls for continual assessment of risk factor and dangers associated with food allergy, keeping abreast of changes in the food industry. Allergologists, regulatory bodies and the food industry are all responsible for food safety. The Allergy Vigilance Network, founded by a university research team and comprising 398 French and Belgian allergologists, has developed a three-point strategy. First, reporting cases of severe anaphylaxis of document allergic origin makes it possible to monitor the prevalence of food allergens and to evaluate the quality of management of allergy-related emergencies, thus providing data suitable for estimating the economic burden of anaphylaxis. The second objective of the network is to set up multicenter trials to determine the prevalence of sensitization to risk allergens, such as peanut, lupin and plant pollen, of which transgenic varieties will soon emerge. The third objective is screening and long-term monitoring of dangers related to new foods, ingredients and adjuvant sensitizing factors. Post-marketing monitoring of potential allergic risks arising from genetically modified food is another aim of the network, together with the establishment of a serum bank, following WHO/FAO recommendations. The Allergy Vigilance Network, together with the French National Institute for Food Safety (AFSSA), the Ministry of Consumer Affairs (DGCCRF) and various patient associations, is striving to analyse and deal with dangers related to the allergenicity of natural and modified food proteins. Publication Types: Comparative Study English Abstract PMID: 18225435 [PubMed - indexed for MEDLINE] 263: Trends Biotechnol. 2008 Mar;26(3):139-45. Epub 2008 Jan 28. Metabolic engineering of carotenoid biosynthesis in plants. Giuliano G, Tavazza R, Diretto G, Beyer P, Taylor MA. Ente per le Nuove tecnologie, l'Energia e l'Ambiente (ENEA), Casaccia Research Center, PO Box 2400, 00123 S.M. di Galeria (Roma), Italy. giuliano@casaccia.enea.it Carotenoids are one of the most diverse classes of natural compounds. Plant carotenoids are composed of a C40 isoprenoid skeleton with or without epoxy, hydroxy and keto groups. They have fundamental roles in human nutrition as antioxidants and vitamin A precursors and their consumption is increasingly associated with protection from a range of diseases. They are also used commercially as safe food, feed and cosmetic colorants and they protect plants from photooxidative stress. In the past six years many metabolic engineering efforts have been undertaken in plants aiming to improve the nutritional value of staple crops, to enable the use of plants as 'cell factories' for producing specialty carotenoids and to improve plant resistance to abiotic stress. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 18222560 [PubMed - indexed for MEDLINE] 264: Ann Allergy Asthma Immunol. 2008 Jan;100(1 Suppl 2):S23-9. Hereditary angiodema: a current state-of-the-art review, VI: novel therapies for hereditary angioedema. Frank MM. Department of Pediatrics, Duke University School of Medicine, Durham, NC 27710, USA. frank007@mc.duke.edu OBJECTIVE: To provide a comprehensive overview on clinical trial design and results of emerging therapies for the treatment of hereditary angioedema (HAE). DATA SOURCES: MEDLINE or PubMed literature searches were conducted to identify double-blind, placebo-controlled trials investigating C1 esterase replacement, kallikrein inhibitor, and bradykinin receptor 2 antagonist therapies. STUDY SELECTION: Ongoing trials or those just recently completed from all companies developing a product for the treatment of HAE are discussed. RESULTS: All of these agents are believed to be effective when tested in patients in phase 1 or phase 2 trials. The studies have many features in common, including being placebo-controlled and blinded; having a preliminary screening visit at which the diagnosis is confirmed; having either low circulating C1 inhibitor protein levels or low levels of functional C1 inhibitor, low C4 levels, and normal C1q levels; enrolling individuals who are relatively early in attacks (4-6 hours from the onset); and stipulating that patients continue taking the medications that they have been taking in the long term. The type of attack acceptable for each treatment protocol varies from study to study. Some allow peripheral edema attacks, some facial attacks, and in some studies, the Food and Drug Administration has allowed purified serum C1 inhibitor to be used as a rescue medication if the patient remains in difficulty after the study drug has been used and found to be ineffective. CONCLUSION: The outlook for new, effective short-term therapy appears to be excellent. In the near future, a whole new therapeutic armamentarium to care for patients with HAE should be available in the United States. Publication Types: Review PMID: 18220149 [PubMed - indexed for MEDLINE] 265: Adv Med Sci. 2007;52:98-103. Food allergies, cross-reactions and agroalimentary biotechnologies. Ronchetti R, Kaczmarski MG, Hałuszka J, Jesenak M, Villa MP. Department of Paediatrics, 2nd School of Medicine, University La Sapienza, S. Andrea Hospital, Rome, Italy. roberto.ronchetti@ospedalesantandrea.it The discrepancy between what the general public and specialist in allergic diseases regard as a true food allergy can in part depend on the frequent evidence of subjects in whom clinical symptoms elicited by a given food allergen are frequently not reproducible: this suggests the existence of allergens variably present in certain foods. In adults and older children common is a form of food allergy associated with inhaled allergens, especially pollens. In this allergic form pollens and various vegetal food often cross react but the underlying scientific rationale is largely unclear. From the study of the "latex-fruits allergic syndrome" and the "oral allergic syndrome" emerged that the cross reactivity depends on epitopes of pollens and vegetables belonging to one of the 14 classes of the "pathogenesis related proteins" (PRPs). Vegetables produce PRPs in response to infection or after plant injury or application of chemicals: long-term conservation and methods used for rapid artificial ripening of vegetables can cause plant to produce PRPs or other allergens. A genetic selection of vegetables "protecting themselves against infection and infestation" by mean of PRPs production is practiced in agroalimentary biotechnology. We deem it urgent that the two realms, Medical Science (Allergology) and Agricultural Biotechnology begin to communicate openly in order to produce food as efficiently as possible but without harming the large part of the population which is predisposed to allergy and react to PRPs. Publication Types: Review PMID: 18217398 [PubMed - indexed for MEDLINE] 266: Proc Natl Acad Sci U S A. 2008 Jan 29;105(4):1339-42. Epub 2008 Jan 23. A Caenorhabditis elegans allatostatin/galanin-like receptor NPR-9 inhibits local search behavior in response to feeding cues. Bendena WG, Boudreau JR, Papanicolaou T, Maltby M, Tobe SS, Chin-Sang ID. Department of Biology, Queen's University, Kingston, ON, Canada. bendenaw@biology.queensu.ca Movement in Caenorhabditis elegans is the result of sensory cues creating stimulatory and inhibitory output from sensory neurons. Four interneurons (AIA, AIB, AIY, and AIZ) are the primary recipients of this information that is further processed en route to motor neurons and muscle contraction. C. elegans has >1,000 G protein-coupled receptors (GPCRs), and their contribution to sensory-based movement is largely undefined. We show that an allatostatin/galanin-like GPCR (NPR-9) is found exclusively in the paired AIB interneuron. AIB interneurons are associated with local search/pivoting behavior. npr-9 mutants display an increased local search/pivoting that impairs their ability to roam and travel long distances on food. With impaired roaming behavior on food npr-9 mutants accumulate more intestinal fat as compared with wild type. Overexpression of NPR-9 resulted in a gain-of-function phenotype that exhibits enhanced forward movement with lost pivoting behavior off food. As such the animal travels a great distance off food, creating arcs to return to food. These findings indicate that NPR-9 has inhibitory effects on the AIB interneuron to regulate foraging behavior, which, in turn, may affect metabolic rate and lipid storage. Publication Types: Research Support, Non-U.S. Gov't PMID: 18216257 [PubMed - indexed for MEDLINE] 267: Toxicology. 2008 Mar 12;245(1-2):24-34. Epub 2007 Dec 17. Immunotoxicological studies of genetically modified rice expressing PHA-E lectin or Bt toxin in Wistar rats. Kroghsbo S, Madsen C, Poulsen M, Schrøder M, Kvist PH, Taylor M, Gatehouse A, Shu Q, Knudsen I. Department of Toxicology and Risk Assessment, National Food Institute, Technical University of Denmark, Mørkhøj Bygade 19, DK-2860 Søborg, Denmark. sck@food.dtu.dk As part of the SAFOTEST project the immunmodulating effect of Cry1Ab protein from Bacillus thuringiensis (Bt) and PHA-E lectin from kidney bean (Phaseolus vulgaris erythroagglutinin) was examined in 28- and 90-day feeding studies in Wistar rats. PHA-E lectin was chosen as positive control. Rats were fed control rice, transgenic rice expressing Cry1Ab protein or PHA-E lectin, or transgenic rice spiked with the purified recombinant protein. Total immunoglobulin levels, mitogen-induced cell proliferation, T-dependent antibody response to sheep red blood cells and the antigen-specific antibody response in serum were examined at the end of the studies. A dose-dependent increase in mesenteric lymph node weight and total immunoglobulin A was seen when feeding PHA-E transgenic rice alone or spiked with 0.1% purified PHA-E lectin for 90 days indicating a local effect of PHA-E in the intestine. No adverse effects of Cry1Ab protein were found. An anti-PHA-E and anti-Cry1Ab antibody response was induced both after inhalation (control groups) and after inhalation/ingestion (groups fed recombinant protein alone or together with transgenic rice). In conclusion, only PHA-E lectin was found to have an immunomodulating effect when feeding rats for 90 days with approximately 70 mg PHA-E/kg bodyweight per day. As both PHA-E lectin and Cry1Ab protein were capable of inducing an antigen-specific antibody response it is important to make careful considerations when designing future animal studies to avoid intake of proteins from the other groups by inhalation as well as to examine the sensitization and elicitation potential of 'foreign' proteins before introduction to the world market. Publication Types: Research Support, Non-U.S. Gov't PMID: 18215453 [PubMed - indexed for MEDLINE] 268: J Food Sci. 2008 Jan;73(1):S64-9. Comparative physicochemical properties and structure of rice containing the sck+cryIAc genes and its nontransgenic counterpart. Li X, Huang K, Zhu B, Liang Z, Wei L, Luo Y. College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, China. The physicochemical properties and structure of an insect-resistant rice, Liangyou Kefeng Nr. 6 (IRR), containing the sck and cryIAc genes were compared with those of its nontransgenic counterpart designated as Liangyou 2186 (control), considering their key role in determining commercial value. Basically the appearance of IRR was not affected in terms of size and shape after foreign gene transformation but improved with lower chalkiness. The milling yield of IRR with lower chalkiness was higher measured by head rice yield compared with its parental control. The differences in appearance and milling quality were confirmed by the structure of raw rice grain by scanning electron microscopy (SEM). Slight differences were observed in pasting properties and textural quality determined by rapid viscosity analyzer and texture analyzer which was in agreement with the result of the structure of cooked rice grain by SEM. The above differences might be as a result of a positional effect of T-DNA insertion. On the whole, the appearance, milling quality, and eating quality of IRR were not adversely affected by transgenes, which will facilitate its acceptance by the consumer after commercialization. Publication Types: Research Support, Non-U.S. Gov't PMID: 18211372 [PubMed - indexed for MEDLINE] 269: Ecotoxicol Environ Saf. 2008 Jun;70(2):327-33. Epub 2008 Feb 21. Does Cry1Ab protein affect learning performances of the honey bee Apis mellifera L. (Hymenoptera, Apidae)? Ramirez-Romero R, Desneux N, Decourtye A, Chaffiol A, Pham-Delègue MH. Instituto de Ecologia A.C., Km. 2.5 Carretera Antigua a Coatepec No. 351 El Haya, 91070 Xalapa, Veracruz, Mexico. Genetically modified Bt crops are increasingly used worldwide but side effects and especially sublethal effects on beneficial insects remain poorly studied. Honey bees are beneficial insects for natural and cultivated ecosystems through pollination. The goal of the present study was to assess potential effects of two concentrations of Cry1Ab protein (3 and 5000 ppb) on young adult honey bees. Following a complementary bioassay, our experiments evaluated effects of the Cry1Ab on three major life traits of young adult honey bees: (a) survival of honey bees during sub-chronic exposure to Cry1Ab, (b) feeding behaviour, and (c) learning performance at the time that honey bees become foragers. The latter effect was tested using the proboscis extension reflex (PER) procedure. The same effects were also tested using a chemical pesticide, imidacloprid, as positive reference. The tested concentrations of Cry1Ab protein did not cause lethal effects on honey bees. However, honey bee feeding behaviour was affected when exposed to the highest concentration of Cry1Ab protein, with honey bees taking longer to imbibe the contaminated syrup. Moreover, honey bees exposed to 5000 ppb of Cry1Ab had disturbed learning performances. Honey bees continued to respond to a conditioned odour even in the absence of a food reward. Our results show that transgenic crops expressing Cry1Ab protein at 5000 ppb may affect food consumption or learning processes and thereby may impact honey bee foraging efficiency. The implications of these results are discussed in terms of risks of transgenic Bt crops for honey bees. Publication Types: Research Support, Non-U.S. Gov't PMID: 18206234 [PubMed - indexed for MEDLINE] 270: Shokuhin Eiseigaku Zasshi. 2007 Dec;48(6):170-8. [Development and evaluation of qualitative detection methods for unapproved genetically modified rice (LLRice)] [Article in Japanese] Watanabe T, Shiramasa Y, Furui S, Kitta K, Minegishi Y, Akiyama H, Maitani T. National Institute of Health Sciences, 1-18-1 Kamiyoga, Setagaya-ku, Tokyo 158-8501, Japan. We developed a specific method to extract DNA from rice grain samples and modified the qualitative real-time PCR method provided by Bayer Co., Ltd. for reliable detection of the genetically modified (GM) rice variety, LLRice601, which has not undergone safety assessment for regulatory approval in Japan. Moreover, we conducted a data analysis to confirm the results obtained with real-time PCR. The yields of DNA extracted from powdered samples of rice grains were almost equal among 5 different varieties of rice, and there was no significant difference in the yield over three days. Reliable results were obtained using 50 ng of the extracted DNA as the template for real-time PCR. To examine the adequacy of the methods, we organized an interlaboratory study with the participation of 2 laboratories, in which 80 test samples were analyzed in a blinded manner. The statistical analysis revealed no significant difference in the Ct value for the endogenous gene of the DNA samples and for the targeted DNA sequence of 0.1% samples. The limit of detection of the method was approximately 0.1%. Analysis of the fluorescence intensity of the PCR-amplified product of the construct-specific DNA sequence suggested that it may be reasonable to judge a sample as positive when a Ct value of less than 40 is obtained. Publication Types: English Abstract Research Support, Non-U.S. Gov't PMID: 18203502 [PubMed - indexed for MEDLINE] 271: Proc Natl Acad Sci U S A. 2008 Feb 5;105(5):1431-5. Epub 2008 Jan 17. Comment in: Proc Natl Acad Sci U S A. 2008 Feb 12;105(6):1777-8. Nutritional impact of elevated calcium transport activity in carrots. Morris J, Hawthorne KM, Hotze T, Abrams SA, Hirschi KD. Vegetable and Fruit Improvement Center, Texas A&M University, College Station, TX 77845, USA. Nutrition recommendations worldwide emphasize ingestion of plant-based diets rather than diets that rely primarily on animal products. However, this plant-based diet could limit the intake of essential nutrients such as calcium. Osteoporosis is one of the world's most prevalent nutritional disorders, and inadequate dietary calcium is a known contributor to the pathophysiology of this condition. Previously, we have modified carrots to express increased levels of a plant calcium transporter (sCAX1), and these plants contain approximately 2-fold-higher calcium content in the edible portions of the carrots. However, it was unproven whether this change would increase the total amount of bioavailable calcium. In randomized trials, we labeled these modified carrots with isotopic calcium and fed them to mice and humans to assess calcium bioavailability. In mice feeding regimes (n = 120), we measured (45)Ca incorporation into bones and determined that mice required twice the serving size of control carrots to obtain the calcium found in sCAX1 carrots. We used a dual-stable isotope method with (42)Ca-labeled carrots and i.v. (46)Ca to determine the absorption of calcium from these carrots in humans. In a cross-over study of 15 male and 15 female adults, we found that when people were fed sCAX1 and control carrots, total calcium absorption per 100 g of carrots was 41% +/- 2% higher in sCAX1 carrots. Both the mice and human feeding studies demonstrate increased calcium absorption from sCAX1-expressing carrots compared with controls. These results demonstrate an alternative means of fortifying vegetables with bioavailable calcium. Publication Types: Randomized Controlled Trial Research Support, N.I.H., Extramural Research Support, U.S. Gov't, Non-P.H.S. PMID: 18202180 [PubMed - indexed for MEDLINE] 272: J AOAC Int. 2007 Nov-Dec;90(6):1517-25. Detection and characterization of cry1Ac transgene construct in Bt cotton: multiple polymerase chain reaction approach. Singh CK, Ojha A, Kachru DN. Industrial Toxicology Research Centre, Post Box No. 80, M.G. Marg Lucknow-226001 U.P., India. To comply with international labeling regulations for genetically modified (GM) crops and food, and to enable proper identification of GM organisms (GMOs), effective methodologies and reliable approaches are needed. The spurious and unapproved GM planting has contributed to crop failures and commercial losses. To ensure effective and genuine GM cultivation, a methodology is needed to detect and identify the trait of interest and concurrently evaluate the structural and functional stability of the transgene insert. A multiple polymerase chain reaction (PCR) approach was developed for detection, identification, and gene stability confirmation of cry1Ac transgene construct in Bt cotton. As many as 9 samples of Bt cotton hybrid seeds comprising 3 approved Bt hybrids, MECH-12Bt, MECH-162Bt, MECH-184Bt, and a batch of 6 nonapproved Bt hybrids were tested. Initially, single standard PCR assays were run to amplify predominant GM DNA sequences (CaMV 35S promoter, nos terminator, and npt-II marker gene); a housekeeping gene, Gossypium hirsutum fiber-specific acyl carrier protein gene (acp1); a trait-specific transgene (cry1Ac); and a sequence of 7S 3' transcription terminator which specifically borders with 3' region of cry1Ac transgene cassette. The concurrent amplification of all sequences of the entire cassette was performed by 3 assays, duplex, triplex, and quadruplex multiplex PCR assays, under common assay conditions. The identity of amplicons was reconfirmed by restriction endonuclease digestion profile. The 2 distinct transgene cassettes, cry1Ac and npt-II, of the Bt cotton were amplified using the respective forward primer of promoter and reverse primer of terminator. The resultant amplicons were excised, eluted, and purified. The purified amplicons served as template for nested PCR assays. The nested PCR runs confirmed the transgene construct orientation and identity. The limit of detection as established by our assay for GM trait (cry1Ac) was 0.1%. This approach can be adopted as a standard procedure for complete molecular characterization of Bt cotton. These assays will be of interest and use to importers, breeders, research laboratories, safety regulators, and food processors for detection of cry1Ac bearing GMOs. Publication Types: Research Support, Non-U.S. Gov't PMID: 18193727 [PubMed - indexed for MEDLINE] 273: Trends Biotechnol. 2008 Feb;26(2):77-81. Epub 2008 Jan 11. Potential for metabolic engineering of resveratrol biosynthesis. Halls C, Yu O. Donald Danforth Plant Science Center, 975 North Warson Road, Saint Louis, MO 63132, USA. Resveratrol, an interesting plant phenolic compound, is found in red wine but is not widely distributed in other common food sources. Health benefits of resveratrol include prevention of cardiovascular diseases and cancers, and--as discovered more recently--promotion of longevity in several animal systems. The pathway and enzymes for resveratrol biosynthesis are well characterized. Furthermore, metabolic engineering of this compound has been achieved in plants, microbes and animals. This review attempts to summarize current understanding of resveratrol pathway-engineering in various systems, to outline the challenges in commercial applications and to identify future opportunities for resveratrol bioengineering. Publication Types: Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Review PMID: 18191264 [PubMed - indexed for MEDLINE] 274: Trends Biotechnol. 2008 Feb;26(2):64-9. Epub 2008 Jan 11. Can GM sorghum impact Africa? Botha GM, Viljoen CD. GMO Testing Facility, University of the Free State, PO Box 339, Bloemfontein 9300, South Africa. bothagm.sci@mail.ufs.ac.za It is said that genetic modification (GM) of grain sorghum has the potential to alleviate hunger in Africa. To this end, millions of dollars have been committed to developing GM sorghum. Current developments in the genetic engineering of sorghum are similar to efforts to improve cassava and other traditional African crops, as well as rice in Asia. On closer analysis, GM sorghum is faced with the same limitations as 'Golden Rice' (GM rice) in the context of combating vitamin A deficiency (VAD) efficiently and sustainably. Thus, it is questionable whether the cost of developing GM sorghum can be justified when compared to the cost of investing in sustainable agricultural practice in Africa. Publication Types: Review PMID: 18191263 [PubMed - indexed for MEDLINE] 275: Nat Biotechnol. 2008 Jan;26(1):73-81. Erratum in: Nat Biotechnol. 2008 Feb;26(2):241. Comment in: Nat Biotechnol. 2008 Oct;26(10):1070-1; author reply 1071-2. Allergenicity assessment of genetically modified crops--what makes sense? Goodman RE, Vieths S, Sampson HA, Hill D, Ebisawa M, Taylor SL, van Ree R. Department of Food Science & Technology, University of Nebraska, Lincoln, Nebraska, 68583-0955, USA. rgoodman2@unlnotes.unl.edu GM crops have great potential to improve food quality, increase harvest yields and decrease dependency on certain chemical pesticides. Before entering the market their safety needs to be scrutinized. This includes a detailed analysis of allergenic risks, as the safety of allergic consumers has high priority. However, not all tests currently being applied to assessing allergenicity have a sound scientific basis. Recent events with transgenic crops reveal the fallacy of applying such tests to GM crops. Publication Types: Research Support, U.S. Gov't, Non-P.H.S. Review PMID: 18183024 [PubMed - indexed for MEDLINE] 276: J Appl Toxicol. 2008 Aug;28(6):734-48. Adverse effect of tannery waste leachates in transgenic Drosophila melanogaster: role of ROS in modulation of Hsp70, oxidative stress and apoptosis. Siddique HR, Gupta SC, Mitra K, Bajpai VK, Mathur N, Murthy RC, Saxena DK, Chowdhuri DK. Embryotoxicology Section, Industrial Toxicology Research Centre, P. O. Box No. 80, M. G. Marg, Lucknow 226 001, Uttar Pradesh, India. Leachate is a complex chemical mixture of chemicals produced as a result of leaching of solid wastes. The potential toxicity of leachates is a major environmental health concern. The present study evaluated the role of ROS in tannery leachates induced Hsp70 expression, antioxidant enzymes and apoptosis in Drosophila. Different concentrations (0.05-2.0%) of leachates prepared from tannery waste at different pH (7.00, 4.93 and 2.88) were mixed with Drosophila food and fed to the larvae for 2-48 h to examine the different stress and apoptotic markers. A concentration- and time-dependent significant increase in Hsp70 expression, ROS generation, antioxidant enzymes activities and MDA content were observed in the exposed larvae. Activities of antioxidant enzymes were delayed compared with Hsp70 expression and MDA level in the exposed organisms. Apoptotic cell death was observed in the exposed larvae at higher concentrations concurrent with a significant regression in Hsp70 along with a higher level of ROS generation. A positive correlation drawn between ROS generation and apoptotic markers and a negative correlation between apoptotic markers and Hsp70 expression at these concentrations indicated the important role of ROS in the induction of cellular damage in the exposed organisms. There was a significant generation of ROS in the larvae exposed to 0.5% of leachates which did not interfere with the protection of their cells by Hsp70 and antioxidant enzymes. However, generation of significantly higher levels of ROS in the larvae exposed to 1.0% and 2.0% leachates may decrease Hsp70 expression thus leading to mitochondria-mediated caspase-dependent apoptotic cell death. Copyright 2008 John Wiley & Sons, Ltd. Publication Types: Research Support, Non-U.S. Gov't PMID: 18172893 [PubMed - indexed for MEDLINE] 277: Biotechnol Prog. 2008 Jan-Feb;24(1):192-201. Epub 2007 Dec 29. A methodical approach to ultra-scale-down of process sequences: application to casein removal from the milk of transgenic animals. Pampel LW, Boushaba R, Titchener-Hooker NJ. Amgen Process & Analytical Sciences, Thousand Oaks, California 91320, USA. Scale-down models of individual operations are widely used in biopharmaceutical process development to obtain information about the performance of production-scale equipment on the basis of inexpensive and efficient laboratory-scale tests, for the purposes of validation or optimization or characterization studies. We have investigated the ability of scale-down models of whole process sequences to provide reliable information for process scale-up from laboratory- to pilot-scales of operation. Using the example of the recovery of a protein from transgenic milk, we have conducted an a priori scale-down analysis of a projected pilot-scale process sequence. A systematic approach was developed to ensure that all critical aspects of process behavior were included in the scale-down model, resulting in the creation of an accurate and reliable scale-down representation of the pilot-scale process. The data from scale-down process trials conducted at 70 and 200 mL scales of operation served to highlight crucial factors determining process performance, and proved reliable in predicting the performance of the pilot-scale process over a scaling factor of 1000. Publication Types: Research Support, Non-U.S. Gov't PMID: 18163641 [PubMed - indexed for MEDLINE] 278: Nat Rev Genet. 2008 Feb;9(2):91-101. Towards a better bowl of rice: assigning function to tens of thousands of rice genes. Jung KH, An G, Ronald PC. Department of Plant Pathology, 1 Shields Avenue, UC Davis, Davis, California 95616, USA. Rice, one of the most important food crops for humans, is the first crop plant to have its genome sequenced. Rice whole-genome microarrays, genome tiling arrays and genome-wide gene-indexed mutant collections have recently been generated. With the availability of these resources, discovering the function of the estimated 41,000 rice genes is now within reach. Such discoveries have broad practical implications for understanding the biological processes of rice and other economically important grasses such as cereals and bioenergy crops. Publication Types: Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Review PMID: 18160965 [PubMed - indexed for MEDLINE] 279: Can J Infect Dis. 2000 May;11(3):142-53. How novel methods can help discover more information about foodborne pathogens. Griffiths MW. Department of Food Science, University of Guelph, Guelph, Ontario. Considerable emphasis is being placed on quantitative risk assessment modelling as a basis for regulation of trade in food products. However, for models to be accurate, information about the behaviour of potential pathogens in foods needs to be available. The question is how to obtain this knowledge in a simple and cost effective way. One technique that has great potential is the use of reporter bacteria which have been genetically modified to express a phenotype that can be easily monitored, such as light production in luminescent organisms. Bacteria carrying these (lux) genes can easily be detected using simple luminometers or more sophisticated low light imaging equipment.By monitoring light output from these bacteria over time, it can easily be determined if the organism is growing (resulting in an increase in light emission), is dead (causing a decrease in light production) or is injured (light output remains constant). The use of imaging systems allows the response of bioluminescent bacteria to be studied directly on the food, making the technique even more useful. Applications of bioluminescence are discussed below and include use as reporters of gene expression; biocide efficacy and antibiotic susceptibility; sub-lethal injury; adhesion and biofilm formation; the microbial ecology of foods; pathogenesis; and as biosensors. PMID: 18159282 [PubMed - in process] 280: BMC Plant Biol. 2007 Dec 20;7:67. TaMSH7: a cereal mismatch repair gene that affects fertility in transgenic barley (Hordeum vulgare L.). Lloyd AH, Milligan AS, Langridge P, Able JA. School of Agriculture, Food & Wine, The University of Adelaide, Waite Campus, PMB1, Glen Osmond, South Australia, 5064, Australia. andrew.lloyd@adelaide.edu.au BACKGROUND: Chromosome pairing, recombination and DNA repair are essential processes during meiosis in sexually reproducing organisms. Investigating the bread wheat (Triticum aestivum L.) Ph2 (Pairing homoeologous) locus has identified numerous candidate genes that may have a role in controlling such processes, including TaMSH7, a plant specific member of the DNA mismatch repair family. RESULTS: Sequencing of the three MSH7 genes, located on the short arms of wheat chromosomes 3A, 3B and 3D, has revealed no significant sequence divergence at the amino acid level suggesting conservation of function across the homoeogroups. Functional analysis of MSH7 through the use of RNAi loss-of-function transgenics was undertaken in diploid barley (Hordeum vulgare L.). Quantitative real-time PCR revealed several T0 lines with reduced MSH7 expression. Positive segregants from two T1 lines studied in detail showed reduced MSH7 expression when compared to transformed controls and null segregants. Expression of MSH6, another member of the mismatch repair family which is most closely related to the MSH7 gene, was not significantly reduced in these lines. In both T1 lines, reduced seed set in positive segregants was observed. CONCLUSION: Results presented here indicate, for the first time, a distinct functional role for MSH7 in vivo and show that expression of this gene is necessary for wild-type levels of fertility. These observations suggest that MSH7 has an important function during meiosis and as such remains a candidate for Ph2. Publication Types: Research Support, Non-U.S. Gov't PMID: 18096080 [PubMed - indexed for MEDLINE] 281: Environ Toxicol Chem. 2008 Jan;27(1):188-95. Toxicological safety assessment of genetically modified Bacillus thuringiensis with additional N-acyl homoserine lactonase gene. Peng D, Zhou C, Chen S, Ruan L, Yu Z, Sun M. State Key Laboratory of Agricultural Microbiology, College of Life Sciences and Technology, Huazhong Agricultural University, Wuhan, People's Republic of China. The aim of the present study is to evaluate the toxicology safety to mammals of a genetically modified (GM) Bacillus thuringiensis with an additional N-acyl homoserine lactones gene (aiiA), which possesses insecticidal activity together with restraint of bacterial pathogenicity and is intended for use as a multifunctional biopesticide. Safety assessments included an acute oral toxicity test and 28-d animal feeding study in Wistar rats, primary eye and dermal irritation in Zealand White rabbits, and delayed contact hypersensitivity in guinea pigs. Tests were conducted using spray-dried powder preparation. This GM product showed toxicity neither in oral acute toxicity test nor in 28-d animal feeding test at a dose of 5,000 mg/kg body weight. During the animal feeding test, there were no significant differences in growth, food and water consumption, hematology, blood biochemical indices, organ weights, and histopathology finding between rats in controls and tested groups. Tested animals in primary eye and dermal irritation and delayed contact hypersensitivity test were also devoid of any toxicity compared to controls. All the above results demonstrated that the GM based multifunctional B. thuringiensis has low toxicity and low eye and dermal irritation and would not cause hypersensitivity to laboratory mammals and therefore could be regarded as safe for use as a pesticide. Publication Types: Research Support, Non-U.S. Gov't PMID: 18092859 [PubMed - indexed for MEDLINE] 282: Toxicol Sci. 2008 Apr;102(2):425-32. Epub 2007 Dec 15. A gene-shuffled glyphosate acetyltransferase protein from Bacillus licheniformis (GAT4601) shows no evidence of allergenicity or toxicity. Delaney B, Zhang J, Carlson G, Schmidt J, Stagg B, Comstock B, Babb A, Finlay C, Cressman RF, Ladics G, Cogburn A, Siehl D, Bardina L, Sampson H, Han Y. Pioneer Hi-Bred International, Inc., Johnston, Iowa 50131, USA. The glyphosate acetyltransferase (gat) gene from Bacillus licheniformis was subjected to multiple rounds of gene shuffling to optimize kinetics of corresponding GAT proteins to acetylate the herbicide active ingredient glyphosate. Genetically modified soybeans expressing the gat4601 gene (356043 soybeans) are tolerant to the application of glyphosate. The current manuscript reports the outcome of the allergenicity and toxicity assessment for the GAT4601 protein. Bioinformatic comparison of the amino acid sequence of GAT4601 did not identify similarities to known allergenic or toxic proteins. In vitro studies conducted with heterologously produced GAT4601 protein demonstrated that it was rapidly degraded in simulated gastric fluid containing pepsin (< 30 s) and in simulated intestinal fluid containing pancreatin (< 2 min) and completely inactivated at temperatures above 56 degrees C. The GAT4601 protein expressed in planta is not glycosylated and similar protein profiles were observed in flour extracts from 356043 soybeans and nontransgenic near isoline comparator soybeans (Jack) using serum from soy allergic persons. No evidence of adverse effects was observed in mice following acute oral exposure to 2000 mg/kg of GAT4601 protein or in a repeated dose dietary exposure study at doses of 800-1000 mg/kg/day. This comprehensive assessment demonstrates that the GAT4601 protein does not present a risk for adverse effects in humans when used in the context of agricultural biotechnology. Publication Types: In Vitro Research Support, Non-U.S. Gov't PMID: 18084044 [PubMed - indexed for MEDLINE] 283: Trends Plant Sci. 2008 Jan;13(1):28-35. Epub 2007 Dec 20. Folate biofortification in food plants. Bekaert S, Storozhenko S, Mehrshahi P, Bennett MJ, Lambert W, Gregory JF 3rd, Schubert K, Hugenholtz J, Van Der Straeten D, Hanson AD. Department of Molecular Genetics, Ghent University, K.L. Ledeganckstraat 35, B-9000 Gent, Belgium. Folate deficiency is a global health problem affecting many people in the developing and developed world. Current interventions (industrial food fortification and supplementation by folic acid pills) are effective if they can be used but might not be possible in less developed countries. Recent advances demonstrate that folate biofortification of food crops is now a feasible complementary strategy to fight folate deficiency worldwide. The genes and enzymes of folate synthesis are sufficiently understood to enable metabolic engineering of the pathway, and results from pilot engineering studies in plants (and bacteria) are encouraging. Here, we review the current status of investigations in the field of folate enhancement on the eve of a new era in food fortification. Publication Types: Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Review PMID: 18083061 [PubMed - indexed for MEDLINE] 284: Pest Manag Sci. 2008 Apr;64(4):479-88. Comparison of herbicide regimes and the associated potential environmental effects of glyphosate-resistant crops versus what they replace in Europe. Kleter GA, Harris C, Stephenson G, Unsworth J. RIKILT-Institute of Food Safety, Wageningen University and Research Centre, Wageningen, Holland. gijs.kleter@wur.nl While cultivation of transgenic crops takes place in seven of the EU member states, this constitutes a relatively limited part of the total acreage planted to these crops worldwide. The only glyphosate-resistant (GR) crop grown commercially until recently has been soybean in Romania. In addition, large-scale experimental European data exist for GR sugar and fodder beets, and, to a lesser extent, GR oilseed rape. These GR crops are likely to have an impact both on the use of herbicides and on the environmental impact of the latter. From the data on these GR crops, it appears that quantities of herbicides applied to GR beets are decreased while those on GR soybean are slightly increased compared with their conventional counterparts. Depending on the parameters used for prediction or measurement of environmental impacts of GR crops, generally similar or less negative impacts were observed compared with conventional crops. Favourable environmental effects of the glyphosate-containing herbicide regimes on GR crops appear feasible, provided appropriate measures for maintaining biodiversity and prevention of volunteers and gene flow are applied. Copyright (c) 2007 Society of Chemical Industry. Publication Types: Comparative Study Research Support, Non-U.S. Gov't Review PMID: 18078305 [PubMed - indexed for MEDLINE] 285: Nature. 2007 Dec 13;450(7172):928-9. Showdown for Europe. Abbott A, Schiermeier Q. Publication Types: News PMID: 18075535 [PubMed - indexed for MEDLINE] 286: Nature. 2007 Dec 13;450(7172):921. Directive action required. [No authors listed] Publication Types: Editorial PMID: 18075528 [PubMed - indexed for MEDLINE] 287: Nat Biotechnol. 2007 Dec;25(12):1356; author reply 1359-60. Comment on: Nat Biotechnol. 2007 Sep;25(9):981-7. GM soybeans--revisiting a controversial format. Cummins J. Publication Types: Comment Letter PMID: 18066023 [PubMed - indexed for MEDLINE] 288: Nat Biotechnol. 2007 Dec;25(12):1356-8. Comment on: Nat Biotechnol. 2007 Sep;25(9):981-7. Response to GM soybeans-revisiting a controversial format. [No authors listed] Publication Types: Comment PMID: 18066022 [PubMed - indexed for MEDLINE] 289: Nat Biotechnol. 2007 Dec;25(12):1355; author reply 1359-60. Comment on: Nat Biotechnol. 2007 Sep;25(9):981-7. GM soybeans--revisiting a controversial format. Ho MW, Saunders PT. Publication Types: Comment Letter PMID: 18066021 [PubMed - indexed for MEDLINE] 290: Nat Biotechnol. 2007 Dec;25(12):1355; author reply 1359-60. Comment on: Nat Biotechnol. 2007 Sep;25(9):981-7. GM soybeans--revisiting a controversial format. Leifert C. Publication Types: Comment Letter PMID: 18066020 [PubMed - indexed for MEDLINE] 291: Nat Biotechnol. 2007 Dec;25(12):1355-6; author reply 1359-60. Comment on: Nat Biotechnol. 2007 Sep;25(9):981-7. GM soybeans--revisiting a controversial format. Heinemann JA, Traavik T. Publication Types: Comment Letter PMID: 18066019 [PubMed - indexed for MEDLINE] 292: Nat Biotechnol. 2007 Dec;25(12):1354-5; author reply 1359-60. Comment on: Nat Biotechnol. 2007 Sep;25(9):981-7. GM soybeans--revisiting a controversial format. John B. Publication Types: Comment Letter PMID: 18066018 [PubMed - indexed for MEDLINE] 293: Nat Biotechnol. 2007 Dec;25(12):1351-4; author reply 1359-60. Comment on: Nat Biotechnol. 2007 Sep;25(9):981-7. GM soybeans--revisiting a controversial format. Ermakova IV. Publication Types: Comment Letter PMID: 18066017 [PubMed - indexed for MEDLINE] 294: Nat Biotechnol. 2007 Dec;25(12):1330. Comment in: Nat Biotechnol. 2008 Apr;26(4):379; discussion 379-80. Another inconvenient truth. In Europe, no one apparently wants to listen if you have good news about genetically modified organisms (GMOs). [No authors listed] Publication Types: Editorial PMID: 18066008 [PubMed - indexed for MEDLINE] 295: Biotechnol Genet Eng Rev. 2007;24:263-79. Production of polyunsaturated fatty acids in transgenic plants. Vrinten P, Wu G, Truksa M, Qiu X. Bioriginal Food and Science Corporation, 102 Melville Street, Saskatoon, Saskatchewan, S7J 0R1, Canada. Publication Types: Review PMID: 18059637 [PubMed - indexed for MEDLINE] 296: Biotechnol Adv. 2008 Mar-Apr;26(2):162-8. Epub 2007 Nov 12. Advances in development of transgenic pulse crops. Eapen S. Nuclear Agriculture and Biotechnology Division, Bhabha Atomic Research Centre, Mumbai-400085, India. eapenhome@yahoo.com It is three decades since the first transgenic pulse crop has been developed. Todate, genetic transformation has been reported in all the major pulse crops like Vigna species, Cicer arietinum, Cajanus cajan, Phaseolus spp, Lupinus spp, Vicia spp and Pisum sativum, but transgenic pulse crops have not yet been commercially released. Despite the crucial role played by pulse crops in tropical agriculture, transgenic pulse crops have not moved out from laboratories to large farm lands compared to their counterparts - 'cereals' and the closely related leguminous oil crop - 'soybean'. The reason for lack of commercialization of transgenic pulse crops can be attributed to the difficulty in developing transgenics with reproducibility, which in turn is due to lack of competent totipotent cells for transformation, long periods required for developing transgenics and lack of coordinated research efforts by the scientific community and long term funding. With optimization of various factors which influence genetic transformation of pulse crops, it will be possible to develop transgenic plants in this important group of crop species with more precision and reproducibility. A translation of knowledge from information available in genomics and functional genomics in model legumes like Medicago truncatula and Lotus japonicus relating to factors which contribute to enhancing crop yield and ameliorate the negative consequences of biotic and abiotic stress factors may provide novel insights for genetic manipulation to improve the productivity of pulse crops. Publication Types: Review PMID: 18055156 [PubMed - indexed for MEDLINE] 297: J Microbiol Biotechnol. 2007 Apr;17(4):681-4. Quantitative analysis of phosphinothricin-N-acetyltransferase in genetically modified herbicide tolerant pepper by an enzyme-linked immunosorbent assay. Shim YY, Shin WS, Moon GS, Kim KH. Saskatoon Research Centre, Agriculture and Agri-Food Canada, 107 Science Place, Saskatoon, Saskatchewan S7N OX2, Canada. An immunoassay method was developed to quantitatively detect phosphinothricin-N-acetyltransferase (PAT) encoded by the Bialaphos resistance (bar) gene in genetically modified (GM) pepper. The histidine-tagged PAT was overexpressed in Escherichia coli M15 (pQE31-bar) and efficiently purified by Ni2+ affinity chromatography. A developed sandwich enzyme-linked immunosorbent assay (S-ELISA) method (detection limit: 0.01 microg/ml) was 100-fold more sensitive than a competitive indirect ELISA (CI-ELISA) method or Western blot analysis in detecting the recombinant PAT. In real sample tests, PAT in genetically modified herbicide-tolerant (GMHT) peppers was successfully quantified [4.9 +/- 0.4 microg/g of sample (n = 6)] by the S-ELISA method. The S-ELISA method developed here could be applied to other GMHT crops and vegetables producing PAT. Publication Types: Research Support, Non-U.S. Gov't PMID: 18051284 [PubMed - indexed for MEDLINE] 298: Science. 2007 Nov 30;318(5855):1417. Plants tolerant of high boron levels. Miwa K, Takano J, Omori H, Seki M, Shinozaki K, Fujiwara T. Biotechnology Research Center, University of Tokyo, Tokyo 113-8657, Japan. Reduced crop productivity due to soils containing toxic levels of boron (B) is a worldwide problem in food production. It is estimated that up to 17% of the barley yield losses in southern Australia are caused by B toxicity. We found that the expression of AtBOR4, an Arabidopsis paralog of BOR1, the first identified boron transporter gene, generates plants that are tolerant of high B levels. BOR4 is a polarly localized borate exporter that enhances B efflux from roots. The present study is a foundation for the improvement of crop productivity in soils containing excess B, which are distributed in arid areas of the world. Publication Types: Research Support, Non-U.S. Gov't PMID: 18048682 [PubMed - indexed for MEDLINE] 299: Proc Natl Acad Sci U S A. 2007 Dec 4;104(49):19631-6. Epub 2007 Nov 28. Delayed leaf senescence induces extreme drought tolerance in a flowering plant. Rivero RM, Kojima M, Gepstein A, Sakakibara H, Mittler R, Gepstein S, Blumwald E. Department of Plant Sciences, University of California, Davis, CA 95616, USA. Drought, the most prominent threat to agricultural production worldwide, accelerates leaf senescence, leading to a decrease in canopy size, loss in photosynthesis and reduced yields. On the basis of the assumption that senescence is a type of cell death program that could be inappropriately activated during drought, we hypothesized that it may be possible to enhance drought tolerance by delaying drought-induced leaf senescence. We generated transgenic plants expressing an isopentenyltransferase gene driven by a stress- and maturation-induced promoter. Remarkably, the suppression of drought-induced leaf senescence resulted in outstanding drought tolerance as shown by, among other responses, vigorous growth after a long drought period that killed the control plants. The transgenic plants maintained high water contents and retained photosynthetic activity (albeit at a reduced level) during the drought. Moreover, the transgenic plants displayed minimal yield loss when watered with only 30% of the amount of water used under control conditions. The production of drought-tolerant crops able to grow under restricted water regimes without diminution of yield would minimize drought-related losses and ensure food production in water-limited lands. Publication Types: Research Support, Non-U.S. Gov't PMID: 18048328 [PubMed - indexed for MEDLINE] 300: J Neuroendocrinol. 2008 Feb;20(2):182-7. Epub 2007 Nov 28. Changes in the brain serotonin satiety system in transgenic rats lacking brain angiotensinogen. Voigt JP, Raasch W, Hörtnagl H, Bader M, Fink H, Jöhren O. University of Nottingham, School of Veterinary Medicine and Science, Sutton Bonington Campus, Sutton Bonington, Leicestershire, UK. peter.voigt@nottingham.ac.uk In transgenic rats, TGR(ASrAOGEN)680, with reduced glial expression of angiotensinogen, changes in brain angiotensinogen are associated with reductions in serotonin (5-HT) content and/or 5-HT metabolism as determined in various brain regions, including the hypothalamus. These rats showed an anxious phenotype upon a first behavioural screen. The present study aimed to extend the search for functional consequences of changes in brain 5-HT with respect to feeding behaviour in these transgenic rats. In feeding experiments, rats were treated with the anorectic drug fenfluramine to probe for functional changes in the serotonergic satiety system. Fenfluramine (0.3 mg/kg, i.p.) reduced food intake in TGR(ASrAOGEN)680 rats whereas the minimal effective dose in wild-type rats was 3 mg/kg, i.p. Although, in the cortex, no differences were apparent in the expression of serotonin 5-HT(1A), 5-HT(1B), 5-HT(2C) receptor and 5-HT transporter mRNAs between TGR(ASrAOGEN)680 and wild-type rats, the expression of mRNAs for the 5-HT(2C) receptor and 5-HT transporter mRNA were significantly higher in the hypothalamus of TGR(ASrAOGEN)680 rats compared to wild-type rats. No differences were found in the mRNA levels for hypothalamic 5-HT(1A) and 5-HT(1B) receptors between TGR(ASrAOGEN)680 and wild-type rats. Taken together, these findings suggest that the transgenic effect on the brain 5-HT system is paralleled by functional changes of the serotonergic feeding system. Publication Types: Research Support, Non-U.S. Gov't PMID: 18047554 [PubMed - indexed for MEDLINE] 301: J Appl Microbiol. 2007 Dec;103(6):2248-57. The heterologous expression of polysaccharidase-encoding genes with oenological relevance in Saccharomyces cerevisiae. van Rensburg P, Strauss ML, Lambrechts MG, Cordero Otero RR, Pretorius IS. Department of Viticulture and Oenology, Institute for Wine Biotechnology, Stellenbosch University, Matieland, South Africa. AIMS: The main objective of this study was to develop polysaccharide-degrading wine strains of Saccharomyces cerevisiae, which are able to improve aspects of wine processing and clarification, as well as colour extraction and stabilization during winemaking. METHODS AND RESULTS: Two yeast expression/secretion gene cassettes were constructed, namely (i) a pectinase gene cassette (pPPK) consisting of the endo-polygalacturonase gene (pelE) from Erwinia chrysanthemi and the pectate lyase gene (peh1) from Erwinia carotovora and (ii) a glucanase/xylanase gene cassette (pEXS) containing the endo-beta-1,4-glucanase gene (end1) from Butyrivibrio fibrisolvens and the endo-beta-1,4-xylanase gene (xynC) from Aspergillus niger. The commercial wine yeast strain, VIN13, was transformed separately with these two gene cassettes and checked for the production of pectinase, glucanase and xylanase activities. Pinot Noir, Cinsaut and Muscat d'Alexandria grape juices were fermented using the VIN13[pPPK] pectinase- and the VIN13[pEXS] glucanase/xylanase-producing transformants. Chemical analyses of the resultant wines indicated that (i) the pectinase-producing strain caused a decrease in the concentration of phenolic compounds in Pinot Noir whereas the glucanase/xylanase-producing strain caused an increase in phenolic compounds presumably because of the degradation of the grape skins; (ii) the glucanase/xylanase-producing strain caused a decrease in wine turbidity, especially in Pinot Noir wine, as well as a clear increase in colour intensity and (iii) in the Muscat d'Alexandria and Cinsaut wines, the differences between the control wines (fermented with the untransformed VIN3 strain) and the wines produced by the two transformed strains were less prominent showing that the effect of these polysaccharide-degrading enzymes is cultivar-dependent. CONCLUSIONS: The recombinant wine yeasts producing pectinase, glucanase and xylanase activities during the fermentation of Pinot Noir, Cinsaut and Muscat d'Alexandria grape juice altered the chemical composition of the resultant wines in a way that such yeasts could potentially be used to improve the clarity, colour intensity and stability and aroma of wine. SIGNIFICANCE AND IMPACT OF THE STUDY: Aspects of commercial-scale wine processing and clarification, colour extraction and stabilization, and aroma enhancement could potentially be improved by the use of polysaccharide-degrading wine yeasts without the addition of expensive commercial enzyme preparations. This offers the potential to further improve the price:quality ratio of wine according to consumer expectations. Publication Types: Research Support, Non-U.S. Gov't PMID: 18045408 [PubMed - indexed for MEDLINE] 302: Trends Biotechnol. 2008 Jan;26(1):4-6. Epub 2007 Nov 26. Novel tomato flavours introduced by plastidial terpenoid pathway engineering. Mollet B, Niederberger P, Pétiard V. Nestlé Research Center, Nestec Ltd., Vers-chez-les-Blanc, CH-1000 Lausanne 26, Switzerland. beat.mollet@rdls.nestle.com Until recently breeding efforts centred on high-yield production while sacrificing flavour and taste quality traits of mass produced food products, such as tomatoes. The recent publication of Davidovich-Rikanati et al. demonstrates the technical feasibility of the genetical engineering of pathways in tomato plants to modify their fruit flavour profile in a proof-of-concept approach. The reported work ranks among an increasing number of reported successful modifications of edible plants with a focus on the benefits to end-consumers. PMID: 18037178 [PubMed - indexed for MEDLINE] 303: J Food Sci. 2007 Nov;72(9):S689-95. Zinc and iron bioavailability of genetically modified soybeans in rats. Martino HS, Martin BR, Weaver CM, Bressan J, Esteves EA, Costa NM. Dept. de Nutrição e Saúde, Univ. Federal de Viçosa, Viçosa-MG 36.570-000, Brazil. The aim of this work was to evaluate zinc and iron bioavailability of UFV-116, a new variety without 2 lipoxygenases, with better taste and flavor than a commercial variety OCEPAR 19, containing all 3 isozymes. To evaluate zinc absorption using 65Zn whole body retention and femur 65Zn uptake, rats were given 3 g of a 65ZnCl2 labeled test meal (0.25 microCi). The 2 varieties were tested at the level of 9 and 30 ppm of zinc as defatted soy flour. Two other groups (control) received egg white as source of protein and ZnS04.H20 as the zinc source. To evaluate iron absorption, using 59Fe whole body retention, animals were given a 3 g 59FeCl3 labeled test meal (0.2 microCi). The 2 varieties were tested at 12 and 25 ppm iron as defatted soy flour. Whole fat soy flour of variety 1 (UFV-116) was higher (P < 0.05) in Ca, K, Mg, phytic acid, and oxalate than variety 2 (OCEPAR-19). No difference was observed among the soybean varieties (P > 0.05) for femur 65Zn retention, at different levels of zinc. However, whole body retention was lower (P < 0.05) for UFV-116 than for OCEPAR-19. Femur 65Zn uptake was correlated with the whole body retention; however, whole body retention was more sensitive. Whole body 59Fe retention from UFV-116 was lower (P < 0.05) than from OCEPAR-19. Zinc and iron bioavailability was lower for UFV-116, possibly due to its higher content of antinutrient factors, especially phytate. Publication Types: Research Support, Non-U.S. Gov't PMID: 18034754 [PubMed - indexed for MEDLINE] 304: J Food Sci. 2007 Nov;72(9):R131-7. Nutritional and safety assessments of foods and feeds nutritionally improved through biotechnology: case studies: executive summary of a task force report by the International Life Sciences Institute, Washington, D.C. International Life Sciences Institute. During the last 2 decades, the public and private sectors have made substantial international research progress toward improving the nutritional value of a wide range of food and feed crops. Nevertheless, significant numbers of people still suffer from the effects of undernutrition. In addition, the nutritional quality of feed is often a limiting factor in livestock production systems, particularly those in developing countries. As newly developed crops with nutritionally improved traits come closer to being available to producers and consumers, we must ensure that scientifically sound and efficient processes are used to assess the safety and nutritional quality of these crops. Such processes will facilitate deploying these crops to those world areas with large numbers of people who need them. This document describes 5 case studies of crops with improved nutritional value. These case studies examine the principles and recommendations published by the Intl. Life Sciences Inst. (ILSI) in 2004 for the safety and nutritional assessment of foods and feeds derived from nutritionally improved crops (ILSI 2004). One overarching conclusion that spans all 5 case studies is that the comparative safety assessment process is a valid approach. Such a process has been endorsed by many publications and organizations, including the 2004 ILSI publication. The type and extent of data that are appropriate for a scientifically sound comparative safety assessment are presented on a case-by-case basis in a manner that takes into account scientific results published since the 2004 ILSI report. This report will appear in the January issue of Comprehensive Reviews in Food Science and Food Safety. Publication Types: Guideline PMID: 18034742 [PubMed - indexed for MEDLINE] 305: Toxicol Sci. 2008 Mar;102(1):100-9. Epub 2007 Nov 21. Differences in allergenic potential of food extracts following oral exposure in mice reflect differences in digestibility: potential approaches to safety assessment. Bowman CC, Selgrade MK. Immunotoxicology Branch, Experimental Toxicology Division, National Health and Environmental Effects Research Laboratory, U.S. Environmental Protection Agency, Research Triangle Park, North Carolina 27711, USA. bowman.christal@epa.gov An animal model for food allergy is needed to assess genetically modified food crops for potential allergenicity. The ideal model must produce allergic antibody (IgE) to proteins differentially according to known allergenicity before being used to accurately identify potential allergens among novel proteins. The oral route is the most relevant for exposure to food antigens, and a protein's stability to digestion is a current risk assessment tool based on this natural route. However, normal laboratory animals do not mount allergic responses to proteins administered orally due to oral tolerance, an immunologic mechanism which specifically suppresses IgE. To circumvent oral tolerance and evoke differential IgE responses to a panel of allergenic and nonallergenic food extracts, female C3H/HeJ mice were exposed subcutaneously or orally with cholera toxin as an adjuvant. All foods elicited IgE by the subcutaneous route. Oral exposure, however, resulted in IgE to allergens (peanut, Brazil nut, and egg white) but not to nonallergens (spinach and turkey), provided that the dose and exposures were limited. Additionally, in vitro digestibility assays demonstrated the presence of digestion-stable proteins in the allergenic food extracts but not in the nonallergenic foods. Our results suggest that the subcutaneous route is inadequate to distinguish allergens from nonallergens, but oral exposure under the appropriate experimental conditions will result in differential allergic responses in accordance with known allergenicity. Moreover, those foods containing digestion-resistant proteins provoke allergic responses in this model, supporting the current use of pepsin resistance in the decision tree for potential allergenicity assessment. Publication Types: Research Support, U.S. Gov't, Non-P.H.S. PMID: 18033772 [PubMed - indexed for MEDLINE] 306: J Ind Microbiol Biotechnol. 2008 Mar;35(3):159-66. Epub 2007 Nov 21. Overexpression of the plg1 gene encoding pectin lyase in Penicillium griseoroseum. Cardoso PG, Ribeiro JB, Teixeira JA, de Queiroz MV, de Araújo EF. Departamento de Biologia/Setor de Microbiologia, Universidade Federal de Lavras, Lavras, MG, Brazil. The pectin lyase (PL) is an industrially important enzyme since it is used for maceration and clarification in the process of fruit juice production in food industries. In order to increase the yields of pectin lyase we cloned the plg1 (pectin lyase 1) from Penicillium griseoroseum gene under the control of the strong constitutive promoter of the glyceraldehyde-3-phosphate dehydrogenase gene (gpdA) and the terminator region of the tryptophan synthetase (trpC) gene from Aspergillus nidulans (plasmid pAN52-Plg1) and transformed this construct into the P. griseoroseum strain PG63. One of the pAN52-Plg1 multi-copy transformants (strain 105) grown in culture medium containing glucose or sugar cane juice showed PL activities of 4,804 or 5,202 U ml(-1) respectively, which represented 57- and 132-fold increases. In addition, the apparent specific activity of PL produced by this strain was much higher than the one observed for a commercial pectinase preparation. Evaluation of the extracellular proteins in the culture supernatant of strain 105 by SDS-PAGE showed the presence of a clear and strong band of approximately 40 kDa that probably corresponds to PL. The enzyme yields reported here demonstrate that the system we developed is able to express pectin lyase at levels comparable to, or exceeding, previously reported data. Publication Types: Research Support, Non-U.S. Gov't PMID: 18030511 [PubMed - indexed for MEDLINE] 307: Plant Biotechnol J. 2008 Apr;6(3):213-25. Epub 2007 Nov 19. Regulating innovative crop technologies in Canada: the case of regulating genetically modified crops. Smyth S, McHughen A. College of Biotechnology, University of Saskatchewan, 51 Campus Drive, Saskatoon, SK, Canada S7N 5A8. stuart.smyth@usask.ca The advent of genetically modified crops in the late 1980s triggered a regulatory response to the relatively new field of plant genetic engineering. Over a 7-year period, a new regulatory framework was created, based on scientific principles that focused on risk mitigation. The process was transparent and deliberately sought the input of those involved in crop development from non-governmental organizations, industry, academia and federal research laboratories. The resulting regulations have now been in place for over a decade, and the resilience of the risk-mitigating regulations is evident as there has been no documented case of damage to either environment or human health. Publication Types: Review PMID: 18028290 [PubMed - indexed for MEDLINE] 308: Proc Natl Acad Sci U S A. 2007 Nov 27;104(48):19150-5. Epub 2007 Nov 19. The transcription factor IDEF1 regulates the response to and tolerance of iron deficiency in plants. Kobayashi T, Ogo Y, Itai RN, Nakanishi H, Takahashi M, Mori S, Nishizawa NK. Laboratory of Plant Biotechnology, Graduate School of Agricultural and Life Sciences, University of Tokyo, 1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan. Iron is essential for most living organisms and is often the major limiting nutrient for normal growth. Plants induce iron utilization systems under conditions of low iron availability, but the molecular mechanisms of gene regulation under iron deficiency remain largely unknown. We identified the rice transcription factor IDEF1, which specifically binds the iron deficiency-responsive cis-acting element IDE1. IDEF1 belongs to an uncharacterized branch of the plant-specific transcription factor family ABI3/VP1 and exhibits the sequence recognition property of efficiently binding to the CATGC sequence within IDE1. IDEF1 transcripts are constitutively present in rice roots and leaves. Transgenic tobacco plants expressing IDEF1 under the control of the constitutive cauliflower mosaic virus 35S promoter transactivate IDE1-mediated expression only in iron-deficient roots. Transgenic rice plants expressing an introduced IDEF1 exhibit substantial tolerance to iron deficiency in both hydroponic culture and calcareous soil. IDEF1 overexpression leads to the enhanced expression of the iron deficiency-induced transcription factor gene OsIRO2, suggesting the presence of a sequential gene regulatory network. These findings reveal cis element/trans factor interactions that are functionally linked to the iron deficiency response. Manipulation of IDEF1 also provides another approach for producing crops tolerant of iron deficiency to enhance food and biomass production in calcareous soils. PMID: 18025467 [PubMed - indexed for MEDLINE] 309: Toxicol Lett. 2007 Dec 10;175(1-3):118-35. Epub 2007 Oct 10. Zero tolerances in food and animal feed -- are there any scientific alternatives? A European point of view on an international controversy. Heberer T, Lahrssen-Wiederholt M, Schafft H, Abraham K, Pzyrembel H, Henning KJ, Schauzu M, Braeunig J, Goetz M, Niemann L, Gundert-Remy U, Luch A, Appel B, Banasiak U, Böl GF, Lampen A, Wittkowski R, Hensel A. Federal Institute for Risk Assessment, Section 55, - Residues of Medicinal Products, Diedersdorfer Weg 1, 12277 Berlin, Germany. bfr@bfr.bund.de A number of zero tolerance provisions are contained in both food and animal feed law, e.g. for chemical substances whose occurrence is not permitted or is directly prohibited in food or animal feed. In the European Union, bans of this kind were introduced to give consumers and animals the greatest possible protection from substances with a possible hazard potential within the intendment of the hazard prevention principles and current precautionary measures. This also applies to substances for which an acceptable daily intake cannot be derived and a maximum residue limit cannot, therefore, be established, e.g. due to missing or inadequate toxicological data. Zero tolerances are also under discussion as trade barriers because their use has triggered numerous legal disputes. This paper draws together the results of an evaluation of alternative risk assessment methods to be used for the risk assessment of substances to which currently only zero tolerances apply. It will demonstrate that, depending on the available toxicological data, a scientifically sound risk assessment may still be possible. In this context, the two concepts - margin of exposure and threshold of toxicological concern - are very promising approaches. Until the scientific and sociopolitical discussions have been completed, it is essential that the principle of zero tolerances be upheld, especially for those substances which may be genotoxic carcinogens. In microbiology, there is no legal room for manoeuvre with regard to food safety criteria established for reasons of consumer health protection on the basis of scientific assessments. PMID: 18024010 [PubMed - indexed for MEDLINE] 310: Toxicol Lett. 2007 Dec 10;175(1-3):82-8. Epub 2007 Oct 7. A rapid and inexpensive method to screen for common foods that reduce the action of acrylamide, a harmful substance in food. Hasegawa K, Miwa S, Tajima T, Tsutsumiuchi K, Taniguchi H, Miwa J. Institute for Biological Function, Chubu University, 1200 Matsumoto, Kasugai 487-8501, Japan. By DNA microarray and protein 2-DE screens for Caenorhabditis elegans genes up-regulated by acrylamide, we selected the gst-4 gene and constructed a gst::gfp fusion gene, which was used to transform C. elegans into a biosensor for acrylamide. This biosensor detects acrylamide as a GFP-expression signal in a dose- and time-dependent manner. When the biosensor was exposed to acrylamide together with commercially available powdered green tea, GFP levels decreased to the control level, suggestive of acrylamide detoxification or prevention of GST induction. The present methodology should be applicable for screening of not only harmful substances but also substances that reduce or counteract their harmfulness or action, with appropriately constructed visible biosensors. Publication Types: Research Support, Non-U.S. Gov't PMID: 18023302 [PubMed - indexed for MEDLINE] 311: Commun Agric Appl Biol Sci. 2007;72(1):177-81. Modelling heterogeneity to estimate the ex ante value of biotechnology innovations. Dillen K, Demont M, Tollens E. Centre for Agricultural and food economics Dept. of land management and economics, K.U. Leuven. PMID: 18018883 [PubMed - indexed for MEDLINE] 312: Trends Plant Sci. 2007 Dec;12(12):548-55. Epub 2007 Nov 19. Transgenic strategies for the nutritional enhancement of plants. Zhu C, Naqvi S, Gomez-Galera S, Pelacho AM, Capell T, Christou P. Universitat de Lleida, Av. Alcalde Rovira Roure, 191, E-25198 Lleida, Spain. The nutrients in the human diet ultimately come from plants. However, all our major food crops lack certain essential vitamins and minerals. Although a varied diet provides adequate nutrition, much of the human population, particularly in developing countries, relies on staple crops, such as rice or maize, which does not provide the full complement of essential nutrients. Malnutrition is a significant public health issue in most of the developing world. One way to address this problem is through the enhancement of staple crops to increase their essential nutrient content. Here, we review the current strategies for the biofortification of crops, including mineral fertilization and conventional breeding but focusing on transgenic approaches which offer the most rapid way to develop high-nutrient commercial cultivars. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 18006362 [PubMed - indexed for MEDLINE] 313: J Food Sci. 2007 Aug;72(6):S420-4. Comparison of nutritional quality between Chinese indica rice with sck and cry1Ac genes and its nontransgenic counterpart. Li X, Huang K, He X, Zhu B, Liang Z, Li H, Luo Y. College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, China. Nutritional assessment of transgenic crops used for human food and animal feed is an important aspect of safety evaluations. An insect-resistant rice (IRR) was generated by the stable insertion of sck, a modified cowpea trypsin inhibitor gene, and cry1Ac, encoding a crystal protein from Bacillus thuringiensis into the genome of a common variety of Chinese indica rice. The composition of the brown and milled rice grain from the resulted IRR line designated Liangyou Kefeng No. 6 was compared with that of the parental rice cultivar Liangyou 2186. Nutrients, including the proximates, amino acids, fatty acids, minerals, and vitamins, were measured. The antinutritive components such as phytic acid, lectin, and trypsin inhibitors were also examined. The data demonstrated that the nutritional quality of both the brown and milled rice grains from the transgenic line was substantially equivalent to that of the nontransgenic counterpart, and measured amounts of nutritional components fell within the range of values reported for other commercial lines. Publication Types: Research Support, Non-U.S. Gov't PMID: 17995700 [PubMed - indexed for MEDLINE] 314: Anal Bioanal Chem. 2008 Jan;390(1):377-87. Epub 2007 Nov 11. Detection and characterization of recombinant DNA expressing vip3A-type insecticidal gene in GMOs--standard single, multiplex and construct-specific PCR assays. Singh CK, Ojha A, Bhatanagar RK, Kachru DN. Industrial Toxicology Research Centre, Post Box No. 80, M. G. Marg, Lucknow, 226001, Uttar Pradesh, India. Vegetative insecticidal protein (Vip), a unique class of insecticidal protein, is now part of transgenic plants for conferring resistance against lepidopteron pests. In order to address the imminent regulatory need for detection and labeling of vip3A carrying genetically modified (GM) products, we have developed a standard single PCR and a multiplex PCR assay. As far as we are aware, this is the first report on PCR-based detection of a vip3A-type gene (vip-s) in transgenic cotton and tobacco. Our assay involves amplification of a 284-bp region of the vip-s gene. This assay can possibly detect as many as 20 natural wild-type isolates bearing a vip3A-like gene and two synthetic genes of vip3A in transgenic plants. The limit of detection as established by our assay for GM trait (vip-s) is 0.1%. Spiking with nontarget DNA originating from diverse plant sources had no inhibitory effect on vip-s detection. Since autoclaving of vip-s bearing GM leaf samples showed no deterioration/interference in detection efficacy, the assay seems to be suitable for processed food products as well. The vip-s amplicon identity was reconfirmed by restriction endonuclease assay. The primer set for vip-s was equally effective in a multiplex PCR assay format (duplex, triplex and quadruplex), used in conjunction with the primer sets for the npt-II selectable marker gene, Cauliflower mosaic virus 35S promoter and nopaline synthetase terminator, enabling concurrent detection of the transgene, regulatory sequences and marker gene. Further, the entire transgene construct was amplified using the forward primer of the promoter and the reverse primer of the terminator. The resultant amplicon served as a template for nested PCR to confirm the construct integrity. The method is suitable for screening any vip3A-carrying GM plant and food. The availability of a reliable PCR assay method prior to commercial release of vip3A-based transgenic crops and food would facilitate rapid and efficient regulatory compliance. Publication Types: Research Support, Non-U.S. Gov't PMID: 17994293 [PubMed - indexed for MEDLINE] 315: Nat Biotechnol. 2007 Nov;25(11):1213-4. GMO quantification in processed food and feed. Weighardt F. Publication Types: Letter PMID: 17989666 [PubMed - indexed for MEDLINE] 316: Toxicol Sci. 2008 Feb;101(2):215-25. Epub 2007 Nov 7. Acrylamide-responsive genes in the nematode Caenorhabditis elegans. Hasegawa K, Miwa S, Isomura K, Tsutsumiuchi K, Taniguchi H, Miwa J. Institute for Biological Function, Chubu University, Kasugai 487-8501, Japan. As acrylamide is a known neurotoxin for many animals and potential carcinogen for humans, it came as a surprise when the Swedish National Food Agency and Stockholm University reported in 2002 that it is formed during the frying or baking of foods. We report here genomic and proteomic analyses on genes and proteins of Caenorhabditis elegans exposed to 500 mg/l acrylamide. Of the 21,120 genes profiled, 409 genes were more than twofold upregulated and 111 genes were downregulated. Upregulated genes included many that encode detoxification enzymes such as glutathione S-transferases (GSTs), uridine diphosphate-glucuronosyl/glucosyl transferases, and short-chain type dehydrogenases but only one cytochrome P450. Subsequent proteomic analysis confirmed the heavy involvement of GSTs. Because of their high expression levels and central roles in acrylamide metabolism, we analyzed the in vivo expression patterns of eight gst genes. Although all encoded GST and were more than twofold upregulated by acrylamide treatment, their expression patterns were varied, and their regulation involved the transcription factor SKN-1 (a C. elegans homolog of Nuclear factor E2-related factors 1 and 2). We then selected the gst-4::gfp-transformed C. elegans to study the detoxification rate of acrylamide and its metabolite glycidimide in living animals. This animal detects acrylamide as a green fluorescence protein (GFP) expression signal in a dose- and time-dependent manner and may prove to be a useful tool not only for rapidly and inexpensively detecting acrylamide, a harmful substance in food, but also for analyzing mechanisms of GST induction by acrylamide and other inducers like oxidative stresses. Publication Types: Research Support, Non-U.S. Gov't PMID: 17989133 [PubMed - indexed for MEDLINE] 317: Med Pregl. 2007 May-Jun;60(5-6):295-8. [Diseases caused by viruses and toxins in biological warfare and bioterrorism] [Article in Serbian] Bojić I, Vukadinov J, Minić S. Specijalisticka ordinacija Dr Bojić Beograd. drbojic@net.yu INTRODUCTION: Viruses and toxins, as well as bacteria and rickettsia can potentially be used as biological weapons in conflicts or in bioterrorism. USE OF BIOLOGICAL WEAPONS: The infection can be acquired by inhalation of aerosols, ingestion of contaminated food or water, or direct contact with the skin or mucosa. Special attention must be given to the possible use of genetically modified agents. CONCLUSION: This paper describes the clinical features of diseases caused hbi viruses (smallpox, hemorrhagic Jever and encephalitis) and toxins (botulinum, staphylococcal enterotoxin B, ricinus toxin and mycotoxins) their diagnosis, treatment, as well as basic preventive measures. Publication Types: English Abstract Review PMID: 17988067 [PubMed - indexed for MEDLINE] 318: Crit Rev Food Sci Nutr. 2007;47(8):721-33. Toxicity studies of genetically modified plants: a review of the published literature. Domingo JL. Laboratory of Toxicology and Environmental Health, School of Medicine, Rovira I Virgili University, San Lorenzo, Reus, Spain. joseluis.domingo@urv.cat According to the information reported by the WHO, the genetically modified (GM) products that are currently on the international market have all passed risk assessments conducted by national authorities. These assessments have not indicated any risk to human health. In spite of this clear statement, it is quite amazing to note that the review articles published in international scientific journals during the current decade did not find, or the number was particularly small, references concerning human and animal toxicological/health risks studies on GM foods. In this paper, the scientific information concerning the potential toxicity of GM/transgenic plants using the Medline database is reviewed. Studies about the safety of the potential use of potatoes, corn, soybeans, rice, cucumber, tomatoes, sweet pepper, peas, and canola plants for food and feed were included. The number of references was surprisingly limited. Moreover, most published studies were not performed by the biotechnology companies that produce these products. This review can be concluded raising the following question: where is the scientific evidence showing that GM plants/food are toxicologically safe? Publication Types: Review PMID: 17987446 [PubMed - indexed for MEDLINE] 319: Regul Toxicol Pharmacol. 2008 Feb;50(1):98-113. Epub 2007 Sep 29. Comparative safety assessment of plant-derived foods. Kok EJ, Keijer J, Kleter GA, Kuiper HA. RIKILT Institute of Food Safety, Bornsesteeg 45, PO Box 230, 6700 AE Wageningen, The Netherlands. Esther.kok@wur.nl The second generation of genetically modified (GM) plants that are moving towards the market are characterized by modifications that may be more complex and traits that more often are to the benefit of the consumer. These developments will have implications for the safety assessment of the resulting plant products. In part of the cases the same crop plant can, however, also be obtained by 'conventional' breeding strategies. The breeder will decide on a case-by-case basis what will be the best strategy to reach the set target and whether genetic modification will form part of this strategy. This article discusses important aspects of the safety assessment of complex products derived from newly bred plant varieties obtained by different breeding strategies. On the basis of this overview, we conclude that the current process of the safety evaluation of GM versus conventionally bred plants is not well balanced. GM varieties are elaborately assessed, yet at the same time other crop plants resulting from conventional breeding strategies may warrant further food safety assessment for the benefit of the consumer. We propose to develop a general screening frame for all newly developed plant varieties to select varieties that cannot, on the basis of scientific criteria, be considered as safe as plant varieties that are already on the market. Publication Types: Comparative Study Research Support, Non-U.S. Gov't Review PMID: 17983697 [PubMed - indexed for MEDLINE] 320: Mol Cells. 2007 Oct 31;24(2):301-6. Overexpression of Arabidopsis homogentisate phytyltransferase or tocopherol cyclase elevates vitamin E content by increasing gamma-tocopherol level in lettuce (Lactuca sativa L.). Lee K, Lee SM, Park SR, Jung J, Moon JK, Cheong JJ, Kim M. School of Agricultural Biotechnology and Center for Agricultural Biomaterials, Seoul National University, Seoul 151-921, Korea. Tocopherols, essential components of the human diet, are synthesized exclusively by photosynthetic organisms. To increase tocopherol content by increasing total flux to the tocopherol biosynthetic pathway, genes encoding Arabidopsis homogentisate phytyltransferase (HPT/V-TE2) and tocopherol cyclase (TC/VTE1) were constitutively overexpressed in lettuce (Lactuca sativa L.). Total tocopherol content of the transgenic plants overexpressing either of the genes was increased by more than 2-fold mainly due to an increase in gamma-tocopherol. However, chlorophyll content in the HPT/VTE2 and TC/VTE1 transgenic lines decreased by up to 20% and increased by up to 35%, respectively (P < 0.01). These results demonstrate that manipulation of the tocopherol biosynthetic pathway can increase or decrease chlorophyll content depending on the gene introduced. Publication Types: Research Support, Non-U.S. Gov't PMID: 17978586 [PubMed - indexed for MEDLINE] 321: J Am Vet Med Assoc. 2007 Nov 1;231(9):1340-2. Comment in: J Am Vet Med Assoc. 2008 Feb 1;232(3):350; author reply 350-1. Envisioning the future of veterinary medicine: the imperative for change in veterinary medical education. Prasse KW, Heider LE, Maccabe AT. Association of American Veterinary Medical Colleges, 1101 Vermont Ave NW, Ste 301, Washington, DC 20005, USA. PMID: 17975989 [PubMed - indexed for MEDLINE] 322: Ecol Appl. 2007 Oct;17(7):2123-35. Effect of pollinator abundance on self-fertilization and gene flow: application to GM Canola. Hoyle M, Hayter K, Cresswell JE. PenTAG, Peninsula Medical School, University of Plymouth, Noy Scott House, Barrack Road, Exeter EX2 5DW, United Kingdom. m.w.hoyle@exeter.ac.uk Cross-pollination from fields of transgenic crops is of great public concern. Although cross-pollination in commercial canola (Brassica napus) fields has been empirically measured, field trials are expensive and do not identify the causes of cross-pollination. Therefore, theoretical models can be valuable because they can provide estimates of cross-pollination at any given site and time. We present a general analytical model of field-to-field gene flow due to the following competing mechanisms: the wind, bees, and autonomous pollination. We parameterize the model for the particular case of field-to-field cross-pollination of genetically modified (GM) canola via the wind and via bumble bees (Bombus spp.) and honey bees (Apis mellifera). We make extensive use of the large data set of bee densities collected during the recent U.K. Farm Scale Evaluations. We predict that canola approaches almost full seed set without pollinators and that autonomous pollination is responsible for > or = 25% of seed set, irrespective of pollinator abundance. We do not predict the relative contribution of bees vs. the wind in landscape-scale gene flow in canola. However, under model assumptions, we predict that the maximum field-to-field gene flow due to bumble bees is 0.04% and 0.13% below the current EU limit for adventitious GM presence for winter- and spring-sown canola, respectively. We predict that gene flow due to bees is approximately 3.1 times higher at 20% compared to 100% male-fertility, and due to the wind, 1.3 times higher at 20% compared to 100% male-fertility, for both winter- and spring-sown canola. Bumble bee-mediated gene flow is approximately 2.7 times higher and wind-mediated gene flow approximately 1.7 times lower in spring-sown than in winter-sown canola, regardless of the degree of male-sterility. The model of cross-pollination due to the wind most closely predicted three previously published observations: field-to-field gene flow is low; gene flow increases with the proportion of plants that are male-sterile; and gene flow is higher in winter- than in spring-sown canola. Our results therefore suggest that the wind, not bees, is the main vector of long-distance gene flow in canola. Publication Types: Research Support, Non-U.S. Gov't PMID: 17974346 [PubMed - indexed for MEDLINE] 323: Mol Ecol. 2008 Mar;17(5):1387-95. Epub 2007 Oct 29. Comment in: Mol Ecol. 2008 Mar;17(5):1167-9. Do escaped transgenes persist in nature? The case of an herbicide resistance transgene in a weedy Brassica rapa population. Warwick SI, Légère A, Simard MJ, James T. Agriculture and Agri-Food Canada, Eastern Cereal and Oilseeds Research Center, Central Experimental Farm, Ottawa, ON, Canada K1A OC6. warwicks@agr.gc.ca The existence of transgenic hybrids resulting from transgene escape from genetically modified (GM) crops to wild or weedy relatives is well documented but the fate of the transgene over time in recipient wild species populations is still relatively unknown. This is the first report of the persistence and apparent introgression, i.e. stable incorporation of genes from one differentiated gene pool into another, of an herbicide resistance transgene from Brassica napus into the gene pool of its weedy relative, Brassica rapa, monitored under natural commercial field conditions. Hybridization between glyphosate-resistant [herbicide resistance (HR)]B. napus and B. rapa was first observed at two Québec sites, Ste Agathe and St Henri, in 2001. B. rapa populations at these two locations were monitored in 2002, 2003 and 2005 for the presence of hybrids and transgene persistence. Hybrid numbers decreased over the 3-year period, from 85 out of approximately 200 plants surveyed in 2002 to only five out of 200 plants in 2005 (St Henri site). Most hybrids had the HR trait, reduced male fertility, intermediate genome structure, and presence of both species-specific amplified fragment length polymorphism markers. Both F(1) and backcross hybrid generations were detected. One introgressed individual, i.e. with the HR trait and diploid ploidy level of B. rapa, was observed in 2005. The latter had reduced pollen viability but produced approximately 480 seeds. Forty-eight of the 50 progeny grown from this plant were diploid with high pollen viability and 22 had the transgene (1:1 segregation). These observations confirm the persistence of the HR trait over time. Persistence occurred over a 6-year period, in the absence of herbicide selection pressure (with the exception of possible exposure to glyphosate in 2002), and in spite of the fitness cost associated with hybridization. PMID: 17971090 [PubMed - indexed for MEDLINE] 324: Int J Toxicol. 2007 Sep-Oct;26(5):389-99. Strategies to evaluate the safety of bioengineered foods. Delaney B. Pioneer Hi-Bred International, Inc., DuPont Agriculture and Nutrition, Johnston, Iowa 50131-0550, USA. bryan.delaney@pioneer.com A number of genetically modified (GM) crops bioengineered to express agronomic traits including herbicide resistance and insect tolerance have been commercialized. Safety studies conducted for the whole grains and food and feed fractions obtained from GM crops (i.e., bioengineered foods) bear similarities to and distinctive differences from those applied to substances intentionally added to foods (e.g., food ingredients). Similarities are apparent in common animal models, route of exposure, duration, and response variables typically assessed in toxicology studies. However, because of differences in the nutritional and physical properties of food ingredients and bioengineered foods and in the fundamental goals of the overall safety assessment strategies for these different classes of substances, there are recognizable differences in the individual components of the safety assessment process. The fundamental strategic difference is that the process for food ingredients is structured toward quantitative risk assessment whereas that for bioengineered foods is structured for the purpose of qualitative risk assessment. The strategy for safety assessment of bioengineered foods focuses on evaluating the safety of the transgenic proteins used to impart the desired trait or traits and to demonstrate compositional similarity between the grains of GM and non-GM comparator crops using analytical chemistry and, in some cases, feeding studies. Despite these differences, the similarities in the design of safety studies conducted with bioengineered foods should be recognized by toxicologists. The current paper reviews the basic principles of safety assessment for bioengineered foods and compares them with the testing strategies applied to typical food ingredients. From this comparison it can be seen that the strategies used to assess the safety of bioengineered foods are at least as robust as that used to assess the safety of typical food ingredients. Publication Types: Review PMID: 17963126 [PubMed - indexed for MEDLINE] 325: J Agric Food Chem. 2007 Nov 28;55(24):9846-9. Epub 2007 Oct 26. Improving zinc content and antioxidant activity in transgenic tomato plants with expression of mouse metallothionein-I by mt-I gene. Sheng J, Liu K, Fan B, Yuan Y, Shen L, Ru B. College of Food Science, China Agricultural University, Beijing, China. Metallothioneins (MTs), as a family of low-molecular-weight, cysteine-rich, and metal-binding proteins, show potential for utilization in functional food. Tomato plants were transformed with gene constructs that contained mt-I encoding the mouse MT-I, similar in sense orientation with the constitutively active double 35S promoter from cauliflower mosaic virus. Three independent transformants, which had copies of the gene in their genomes, were obtained. In these transgenic lines, high-level expression of MT-I, high zinc content, and some antioxidant enzyme activities were detected in leaves. The average zinc content in transgenic tomato leaves was 32.7 mg/100 g FW, which about 1.6 times higher than that in wild-type. The superoxide dismutase activity was also higher (68.6, 66.9, and 66.1 U/g FW in the three transformants) than that in wild-type (57.4 U/g FW). In particular, the levels of superoxide free radical scanvenging in the three transformants were 14.2%, 14.6%, and 13.7%, respectively, which about 1.5 times higher than that in control (5.6%). Transgenic MT tomato may potentially be used as an antioxidant and for zinc supplementation. Publication Types: Research Support, Non-U.S. Gov't PMID: 17960876 [PubMed - indexed for MEDLINE] 326: Risk Anal. 2007 Aug;27(4):935-46. An empirical test of competing theories of hazard-related trust: the case of GM food. Allum N. Department of Sociology, University of Surrey, Guildford, UK. n.allum@surrey.ac.uk Few scholars doubt the importance of trust in explaining variation in public perception of technological risk. Relatively little, however, is known about the particular types of judgments that people use in granting or withholding trust. This article presents findings from an empirical study that explores several dimensions of trust relevant for citizens' judgments of scientists involved in the development of GM food. The relationship between particular dimensions of trust and perceptions of GM food risk is also explored, using structural equation modeling. Results suggest that trust judgments based on the perception of shared values are most important in relation to GM food risk, but that judgments about scientists' technical competence are also important. PMID: 17958502 [PubMed - indexed for MEDLINE] 327: Plant Biotechnol J. 2008 Jan;6(1):2-12. Epub 2007 Oct 23. US regulatory system for genetically modified [genetically modified organism (GMO), rDNA or transgenic] crop cultivars. McHughen A, Smyth S. Department of Botany and Plant Sciences, University of California, Riverside, CA 92521-0124, USA. alanmc@ucr.edu This paper reviews the history of the federal regulatory oversight of plant agricultural biotechnology in the USA, focusing on the scientific and political forces moulding the continually evolving regulatory structure in place today. Unlike most other jurisdictions, the USA decided to adapt pre-existing legislation to encompass products of biotechnology. In so doing, it established an overarching committee (Office of Science and Technology Policy) to study and distribute various regulatory responsibilities amongst relevant agencies: the Food and Drug Administration, Environmental Protection Agency and US Department of Agriculture. This paper reviews the history and procedures of each agency in the execution of its regulatory duties and investigates the advantages and disadvantages of the US regulatory strategy. Publication Types: Historical Article Review PMID: 17956539 [PubMed - indexed for MEDLINE] 328: J AOAC Int. 2007 Sep-Oct;90(5):1513-6. Development of agricultural biotechnology and biosafety regulations used to assess the safety of genetically modified crops in Iran. Mousavi A, Malboobi MA, Esmailzadeh NS. National Institute of Genetic Engineering and Biotechnology, P.O. Box 14155-6343, Tehran, 1417863171, Iran. m-amir@nrcgeb.ac.ir Rapid progress in the application of biotechnological methodologies and development of genetically modified crops in Iran necessitated intensive efforts to establish proper organizations and prepare required rules and regulations at the national level to ensure safe application of biotechnology in all pertinent aspects. Practically, preparation of a national biotechnology strategic plan in the country coincided with development of a national biosafety framework that was the basis for the drafted biosafety law. Although biosafety measures were observed by researchers voluntarily, the establishment of national biosafety organizations since the year 2000 built a great capacity to deal with biosafety issues in the present and future time, particularly with respect to food and agricultural biotechnology. Publication Types: Review PMID: 17956001 [PubMed - indexed for MEDLINE] 329: J AOAC Int. 2007 Sep-Oct;90(5):1508-12. Development of agribiotechnology and biosafety regulations used to assess safety of genetically modified crops in Bangladesh. Nasiruddin KM, Nasim A. Bangladesh Agricultural University, Biotechnology Department, Mymensingh 2202, Bangladesh. nasirbiotech@yahoo.com Bangladesh is on the verge of adopting genetically modified (GM) crops for commercial cultivation and consumption as feed and food. Most of the laboratories are engaged in tissue culture and molecular characterization on plants, whereas some have started living modified organism research with shortages of trained manpower, infrastructure, and funding. Nutritionally improved Golden Rice, biotech brinjal, and late blight-resistant potato are in contained trials in a greenhouse, and potato ring spot virus-resistant papaya is in the process of approval for a field trial. The government has taken some initiative in support of GM organism research, which include the formation of a Biotechnology Department in all institutes and the formation of the apex body, the National Task Force Committee on Biotechnology of Bangladesh under the chairpersonship of the Prime Minister. Biosafety policy guidelines and related aspects of biotechnology issues have been approved, and the laws are in the process of being promulgated. Being a party to the Cartagena Protocol, proper biosafety measures are regulated by the appropriate authority as stated. Although there are no laws made yet directly for biosafety of GM crops/foods, the relevant laws on agriculture, medicine, food, import, trade, environment, etc. may suffice and explain the situation. Publication Types: Review PMID: 17956000 [PubMed - indexed for MEDLINE] 330: J AOAC Int. 2007 Sep-Oct;90(5):1500-7. Development of agriculture biotechnology in Pakistan. Zafar Y. Pakistan Atomic Energy Commission, Agriculture and Biotechnology Division, PO Box No. 1114, Islamabad, Pakistan. y_zafar@yahoo.com Agriculture plays an important role in the national economy of Pakistan, where most of the rapidly increasing population resides in rural areas and depends on agriculture for subsistence. Biotechnology has considerable potential for promoting the efficiency of crop improvement, food production, and poverty reduction. Use of modern biotechnology started in Pakistan since 1985. Currently, there are 29 biotech centers/institutes in the country. However, few centers have appropriate physical facilities and trained manpower to develop genetically modified (GM) crops. Most of the activities have been on rice and cotton, which are among the top 5 crops of Pakistan. Biotic (virus/bacterial/insect) and abiotic (salt) resistant and quality (male sterility) genes have already been incorporated in some crop plants. Despite acquiring capacity to produce transgenic plants, no GM crops, either produced locally or imported, have been released in the country. Pakistan is signatory to the World Trade Organization, Convention on Biological Diversity, and Cartagena protocols. Several legislations under the Agreement on Trade-Related Aspects of Intellectual Property Rights have been promulgated in the country. National Biosafety Guidelines have been promulgated in April 2005. The Plant Breeders Rights Act, Amendment in Seed Act-1976, and Geographical Indication for Goods are still passing through discussion, evaluation, and analysis phases. Meanwhile, an illegal GM crop (cotton) has already sneaked into farmer's field. Concerted and coordinated efforts are needed among various ministries for implementation of regulation and capacity building for import/export and local handling of GM crops. Pakistan could easily benefit from the experience of Asian countries, especially China and India, where conditions are similar and the agriculture sector is almost like that of Pakistan. Thus, the exchange of information and experiences is important among these nations. PMID: 17955999 [PubMed - indexed for MEDLINE] 331: J AOAC Int. 2007 Sep-Oct;90(5):1492-9. Application of current allergy assessment guidelines to next-generation biotechnology-derived crops. Bannon GA, Martino-Catt S. Monsanto Co., Global Regulatory Sciences, 800 N. Lindbergh Blvd, St. Louis, MO 63167, USA. gary.a.bannan@monsanto.com In any single day, our immune systems are exposed to thousands of different proteins from the environment and the food we eat. In a portion of the human population, some of those proteins will stimulate the immune systems to synthesize immunoglobulin E in an allergenic response. The discrepancy between the vast numbers of proteins we encounter and the limited number of proteins that actually become allergens have led scientists on a quest to discover what unique features exist that make proteins destined to be allergens. The information gained from these studies has led to an allergy assessment strategy that characterizes the potential allergenicity of biotechnology products prior to their commercialization. This testing strategy appears to be effective as shown by the fact that there have been no clinically documented food allergic reactions to any of the biotechnology proteins introduced into food crops, to date. The next generation of biotechnology products will most likely contain more complex traits, including nutritionally enhanced food crops, and the question arises as to whether the current allergy assessment strategy will be sufficient to protect the health of the consuming public. In this paper, we discuss general allergen characteristics in order to better understand how proteins become allergens, summarize the current allergy assessment process, evaluate the different aspects of this process for their adequacy in determining the allergenic potential of engineered functional foods, and, finally, we assess the possibility of new technologies having a positive impact on the allergy assessment of nutritionally enhanced crops. Publication Types: Review PMID: 17955998 [PubMed - indexed for MEDLINE] 332: J AOAC Int. 2007 Sep-Oct;90(5):1480-91. An overview of methods for assessment of iron bioavailability from foods nutritionally enhanced through biotechnology. Cockell KA. Health Canada, Nutrition Research Division, Food Directorate, 2203C Banting Research Centre, 251 Sir Frederick Banting Driveway, Ottawa, ON, Canada. kevin_cockell@hc-sc.gc.ca Iron deficiency and iron deficiency anemia continue to be significant public health problems worldwide. While supplementation and fortification have been viable means to improve iron nutriture of the population in developed countries, they may be less successful in developing regions for a number of reasons, including complexities in distribution and consumer compliance. Biofortification of staple crops, through conventional plant breeding strategies or modern methods of biotechnology, provides an alternative approach that may be more sustainable once initial investments have been made. Three types of biofortification strategies are being essayed, singly or in combination: increasing the total iron content of edible portions of the plant, decreasing the levels of inhibitors of iron absorption, and increasing the levels of factors that enhance iron absorption. Bioavailability is a key concept in iron nutrition, particularly for nonheme iron such as is found in these biofortified foods. An overview is presented of methods for evaluation of iron bioavailability from foods nutritionally enhanced through biotechnology. PMID: 17955997 [PubMed - indexed for MEDLINE] 333: J AOAC Int. 2007 Sep-Oct;90(5):1470-9. Nutritional and safety assessments of foods and feeds nutritionally improved through biotechnology: lysine maize as a case study. Glenn KC. Monsanto Co., 800 North Lindbergh Blvd, E3NB, St. Louis, MO 63167, USA. kevin.c.glenn@monsanto.com During the last decade, the area of biotech crops modified for agronomic input traits (e.g., herbicide tolerance and insect protection) has increased to 90 million halyear, grown by over 8 million farmers in a total of 17 countries. As adoption of these improved agronomic trait biotech crops has grown, so has interest in biotech crops that have improved nutritional characteristics for use as feed and food. A previous publication by the International Life Sciences Institute (ILSI) reported on the principles and concepts proposed for the nutritional and safety assessments of foods and feeds nutritionally improved through biotechnology. In this paper, the guidelines and principles recommended in the earlier publication are discussed relative to a specific case study, Lysine maize. Lysine maize is a feed ingredient with enhanced nutritional characteristics for poultry and swine and provides an alternative to the need for addition of supplemental lysine to some diets for these animals. The 2004 Task Force of the ILSI has also applied the concepts from that report to 4 other case studies: sweet potato enriched in provitamin A (2 examples, one using biotechnology and one using conventional breeding); Golden Rice 2; double-embryo maize; and ASP-1 enhanced protein sweet potato. PMID: 17955996 [PubMed - indexed for MEDLINE] 334: J AOAC Int. 2007 Sep-Oct;90(5):1445-9. Delivering golden rice to developing countries. Mayer JE. Golden Rice Project, Campus Technologies Freiburg, 79104 Freiburg, Germany. jorge.mayer@goldenrice.org Micronutrient deficiencies create a vicious circle of malnutrition, poverty, and economic dependency that we must strive to break. Golden Rice offers a sustainable solution to reduce the prevalence of vitamin A deficiency-related diseases and mortality, a problem that affects the health of millions of children in all developing countries. The technology is based on the reconstitution of the carotenoid biosynthetic pathway by addition of 2 transgenes. The outcome of this high-tech approach will be provided to end users as nutrient-dense rice varieties that are agronomically identical to their own, locally adapted varieties. This intervention has the potential to reach remote rural populations without access to fortification and supplementation programs. As part of our delivery strategy, we are partnering with government and nongovernment, national and international agricultural institutions to navigate through cumbersome and expensive regulatory regimes that affect the release of genetically modified crops, and to create local demand for the biofortified rice varieties. Publication Types: Review PMID: 17955992 [PubMed - indexed for MEDLINE] 335: J AOAC Int. 2007 Sep-Oct;90(5):1440-4. Impact of foods nutritionally enhanced through biotechnology in alleviating malnutrition in developing countries. Gilani GS, Nasim A. Nutrition Research Division, Food Directorate, Health Products and Food Branch, Health Canada, Government of Canada, 251 Sir Frederick Banting Dwy, Ottawa, ON, Canada. Sarwar_Gilani@hc-sc.gc.ca According to United Nations (UN) projections, the world's population will grow from 6.1 billion in 2000 to 8 billion in 2025 and 9.4 billion in 2050. Most (93%) of the increase will take place in developing countries. The rapid population growth in developing countries creates major challenges for governments regarding food and nutrition security. According to current World Health Organization estimates, more than 3 billion people worldwide, especially in developing countries, are malnourished in essential nutrients. Malnutrition imposes severe costs on a country's population due to impaired physical and cognitive abilities and reduced ability to work. Little progress has been made in improving malnutrition over the past few decades. The Food and Agriculture Organization of the UN would like to see more nutrient-rich foods introduced into these countries, because supplements are expensive and difficult to distribute widely. Biofortification of staple crops through modern biotechnology can potentially help in alleviating malnutrition in developing countries. Several genetically modified crops, including rice, potatoes, oilseeds, and cassava, with elevated levels of essential nutrients (such as vitamin A, iron, zinc, protein and essential amino acids, and essential fatty acids); reduced levels of antinutritional factors (such as cyanogens, phytates, and glycoalkaloid); and increased levels of factors that influence bioavailability and utilization of essential nutrients (such as cysteine residues) are advancing through field trial stage and regulatory processes towards commercialization. The ready availability and consumption of the biofortified crops would have a significant impact in reducing malnutrition and the risk of chronic disease in developing countries. Publication Types: Review PMID: 17955991 [PubMed - indexed for MEDLINE] 336: Food Chem Toxicol. 2008 Jan;46(1):9-33. Epub 2007 Sep 14. The application of post-market monitoring to novel foods. Hepburn P, Howlett J, Boeing H, Cockburn A, Constable A, Davi A, de Jong N, Moseley B, Oberdörfer R, Robertson C, Wal JM, Samuels F. Unilever, Safety and Environmental Assurance Centre, Colworth Park, Sharnbrook, Befordshire MK44 1LQ, United Kingdom. The role of post-market monitoring (PMM) in the safety assessment of novel foods is critically discussed in order to derive guidelines as to in which situations the application of PMM might be warranted. Available data sources on food consumption and health status, and the methodologies for generating such data are reviewed. The paper suggests improvements to make them more applicable for PMM purposes. It is concluded that any PMM programme must be a hypothesis-driven scientific exercise. PMM can have a role as a complement to, but not as a replacement for, a comprehensive pre-market safety assessment. Its use may be appropriate to confirm that product use is as predicted in the pre-market assessment; to provide reassurance that effects observed in the pre-market assessment occur with no greater frequency or intensity in the post-market phase than anticipated; and to investigate the significance of any adverse effects reported by consumers after market-launch. However PMM is insufficiently powerful to test the hypothesis that any effects seen in the pre-market assessment are absent in the post-market phase. Current methodologies place limitations on what PMM can achieve. PMM should only be used when triggered by or when the focus is on specific evidence-based questions. Publication Types: Review PMID: 17950974 [PubMed - indexed for MEDLINE] 337: Appetite. 2008 Mar-May;50(2-3):340-52. Epub 2007 Sep 18. Consumer responses to communication about food risk management. van Dijk H, Houghton J, van Kleef E, van der Lans I, Rowe G, Frewer L. Marketing and Consumer Behaviour Group, Department of Social Sciences, Wageningen University, Hollandseweg 1, 6706 KN Wageningen, The Netherlands. Heleen.vanDijk@wur.nl Recent emphasis within policy circles has been on transparent communication with consumers about food risk management decisions and practices. As a consequence, it is important to develop best practice regarding communication with the public about how food risks are managed. In the current study, the provision of information about regulatory enforcement, proactive risk management, scientific uncertainty and risk variability were manipulated in an experiment designed to examine their impact on consumer perceptions of food risk management quality. In order to compare consumer reactions across different cases, three food hazards were selected (mycotoxins on organically grown food, pesticide residues, and a genetically modified potato). Data were collected from representative samples of consumers in Germany, Greece, Norway and the UK. Scores on the "perceived food risk management quality" scale were subjected to a repeated-measures mixed linear model. Analysis points to a number of important findings, including the existence of cultural variation regarding the impact of risk communication strategies-something which has obvious implications for pan-European risk communication approaches. For example, while communication of uncertainty had a positive impact in Germany, it had a negative impact in the UK and Norway. Results also indicate that food risk managers should inform the public about enforcement of safety laws when communicating scientific uncertainty associated with risks. This has implications for the coordination of risk communication strategies between risk assessment and risk management organizations. Publication Types: Multicenter Study Research Support, Non-U.S. Gov't PMID: 17945386 [PubMed - indexed for MEDLINE] 338: Crit Rev Food Sci Nutr. 2007;47(7):675-99. A review on tomato authenticity: quality control methods in conjunction with multivariate analysis (chemometrics). Arvanitoyannis IS, Vaitsi OB. University of Thessaly School of Agricultural Sciences Department of Agriculture Animal Production & Aquatic Production, Volos, Hellas, Greece. parmenion@uth.gr Authenticity and traceability have been two of the most important issues in the food chain. Authenticity in particular, is closely related with both food quality and safety issues. Vegetables stand for a category of foods heavily affected by adulteration either in terms of geographic origin (national or international level) or production methods (organic or conventional production, fertilizers, pesticides, genetically modified vegetables). This review aims at addressing most of the currently applied methods for ensuring quality control of vegetables; a) instrumental: ion chromatography, high pressure liquid chromatography, atomic absorption spectrophotometry, electronic nose and mass spectroscopy and b) sensory analysis. The results of all the above mentioned methods were analyzed by means of multivariate analysis (principal component analysis, discriminant analysis, cluster analysis, canonical analysis, and factor analysis). All ensuing results and conclusions are summarized in eight comprehensive tables. Publication Types: Review PMID: 17943497 [PubMed - indexed for MEDLINE] 339: J Int Bioethique. 2006 Dec;17(4):109-14. GMOs and development. DaSilva EJ. Section of Life Sciences Division of Basic and Engineering Sciences, UNESCO, Paris, France. PMID: 17939274 [PubMed - indexed for MEDLINE] 340: Nat Biotechnol. 2007 Nov;25(11):1277-9. Epub 2007 Oct 14. Folate fortification of rice by metabolic engineering. Storozhenko S, De Brouwer V, Volckaert M, Navarrete O, Blancquaert D, Zhang GF, Lambert W, Van Der Straeten D. Unit Plant Hormone Signalling and Bio-imaging, Department of Molecular Genetics, Ghent University, K.L. Ledeganckstraat 35, B-9000 Ghent, Belgium. Rice, the world's major staple crop, is a poor source of essential micronutrients, including folates (vitamin B9). We report folate biofortification of rice seeds achieved by overexpressing two Arabidopsis thaliana genes of the pterin and para-aminobenzoate branches of the folate biosynthetic pathway from a single locus. We obtained a maximal enhancement as high as 100 times above wild type, with 100 g of polished raw grains containing up to four times the adult daily folate requirement. Publication Types: Research Support, Non-U.S. Gov't PMID: 17934451 [PubMed - indexed for MEDLINE] 341: Appl Environ Microbiol. 2007 Dec;73(24):8012-7. Epub 2007 Oct 12. Effect of feeding cows genetically modified maize on the bacterial community in the bovine rumen. Wiedemann S, Gürtler P, Albrecht C. Institute of Biochemistry and Molecular Medicine, University of Bern, Buehlstr. 28, CH-3012 Bern, Switzerland. christiane.albrecht@mci.unibe.ch Rumen-cannulated cows (n = 4) were fed successively silage made from either conventional or genetically modified (GM) maize. Results revealed no effects of GM maize on the dynamics of six ruminal bacterial strains (investigated by real-time PCR) compared to the conventional maize silage. Publication Types: Comparative Study Research Support, Non-U.S. Gov't PMID: 17933942 [PubMed - indexed for MEDLINE] 342: Electrophoresis. 2007 Nov;28(22):4247-54. Ultra-fast simultaneous analysis of genetically modified organisms in maize by microchip electrophoresis with LIF detector. Kumar KS, Kang SH. Department of Chemistry and Research Institute Basic Science, Chonbuk National University, Jeonju, South Korea. This study examined the potential of microchip electrophoresis (ME) with a LIF detector using a programmed field strength gradient (PFSG) in a conventional glass double-T microchip for the ultra-fast detection and simultaneous analysis of genetically modified (GM) maize. The separation efficiency and sensitivity at various sieving gels (poly(ethylene oxide) (PEO, M(r) 8,000,000) and 2-hydroxyethylcellulose (HEC) (M(r) 250,000)) and fluorescent dye concentrations were investigated. The PCR products of both the GM and non-GM maize were analyzed within 30 s under the PFSG (470.6 V/cm for 20 s, 117.6 V/cm for 12 s, and 470.6 V/cm for 30 s) with a 2.5% HEC sieving matrix in the running buffer, 1 x Tris-borate EDTA (TBE) (pH 8.30) and 0.5 ppm ethidium bromide. The five transgenic maize varieties (Event176, MON810, Bt11, GA21, and T25) examined in this study were also clearly differentiated by ME-PFSG within 30 s in a single run without any loss of resolution. The ME-PFSG technique is a powerful tool for the ultra-fast detection and simultaneous analysis of GMOs in a variety of foods including maize. Publication Types: Research Support, Non-U.S. Gov't PMID: 17932874 [PubMed - indexed for MEDLINE] 343: Transgenic Res. 2008 Aug;17(4):665-77. Epub 2007 Oct 12. A T7-driven silencing system in transgenic plants expressing T7 RNA polymerase is a nuclear process. Peretz Y, Levy M, Avisar E, Edelbaum O, Rabinowitch H, Sela I. Robert H. Smith Institute for Plant Sciences and Genetics in Agriculture, The Hebrew University of Jerusalem, Faculty of Agricultural, Food and Environmental Quality Sciences, Rehovot, Israel. We previously demonstrated a case of silencing in transgenic plants expressing T7 RNA polymerase in which expression of a reporter gene placed under the control of the T7 promoter was silenced. Here we demonstrate that endogenous genes can be silenced by the same system. The T7-driven silencing system does not conform to several aspects characteristic of post-transcriptional RNA silencing in plants, and this prompted an investigation into the mechanisms underlying this type of silencing. The present paper demonstrates that T7-driven silencing is a post-transcriptional process that is restricted to the nucleus. Nuclear run-on assays indicated the presence of silenced gene transcripts in both orientations. SiRNA corresponding to the silenced gene could not be traced in the cytoplasm but was found in nuclei. The silenced gene was hypermethylated. We present evidence that a tobacco RNA-dependent RNA polymerase (RdRP) is not involved in T7-mediated silencing, but indicate the involvement of a nuclear RdRP in this type of silencing. PMID: 17932780 [PubMed - indexed for MEDLINE] 344: Plant Foods Hum Nutr. 2007 Dec;62(4):185-91. Epub 2007 Oct 11. Towards generating caffeine-free tea by metabolic engineering. Yadav SK, Ahuja PS. Biotechnology Division, Institute of Himalayan Bioresource Technology, CSIR, Palampur, H.P. 176061, India. skyt@rediffmail.com Tea is a rich source of antioxidants which are contributing substantially to the promotion of health and the prevention of various chronic diseases. Despite the fact that tea has various important compounds, it also contains a purine alkaloid, caffeine. High intake of tea leads to an increase in level of caffeine in addition to its important antioxidant constituents. Increased level of caffeine causes several health related problems. Therefore, tea can become a most useful source of beneficial compounds, if only its caffeine level is either decreased or eliminated all together from the plant itself. This could be achieved through either of the techniques; overexpressing caffeine degradative pathway genes or silencing caffeine biosynthesis pathway gene. The identification and cloning of caffeine biosynthesis in tea and degradative genes in microorganisms opens up the possibility of using genetic engineering to produce naturally decaffeinated tea. Here we review these different strategies which can be employed to make caffeine-free tea, a human health beneficial drink. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 17929169 [PubMed - indexed for MEDLINE] 345: BMC Bioinformatics. 2007 Oct 9;8:375. Computational analysis of the relationship between allergenicity and digestibility of allergenic proteins in simulated gastric fluid. Jiang B, Qu H, Hu Y, Ni T, Lin Z. College of Life Sciences, National Lab of Protein Engineering and Genetic Engineering of Plants, Peking University, Beijing 100871, PR China. jiangbingjun@gmail.com BACKGROUND: Safety assessment of genetically modified (GM) food, with regard to allergenic potential of transgene-encoded xenoproteins, typically involves several different methods, evaluation by digestibility being one thereof. However, there are still debates about whether the allergenicity of food allergens is related to their resistance to digestion by the gastric fluid. The disagreements may in part stem from classification of allergens only by their sources, which we believe is inadequate, and the difficulties in achieving identical experimental conditions for studying digestion by simulated gastric fluid (SGF) so that results can be compared. Here, we reclassify allergenic food allergens into alimentary canal-sensitized (ACS) and non-alimentary canal-sensitized (NACS) allergens and use a computational model that simulates gastric fluid digestion to analyze the digestibilities of these two types. RESULTS: The model presented in this paper is as effective as SGF digestion experiments, but more stable and reproducible. On the basis of this model, food allergens are satisfactorily classified as ACS and NACS types by their pathways for sensitization; the former are relatively resistant to gastric fluid digestion while the later are relatively labile. CONCLUSION: The results suggest that it is better to classify allergens into ACS and NACS types when understanding the relationship between their digestibility and allergenicity and the digestibility of a target foreign protein is a parameter for evaluating its allergenicity during safety assessments of GM food. Publication Types: Research Support, Non-U.S. Gov't PMID: 17922925 [PubMed - indexed for MEDLINE] 346: FEBS J. 2007 Nov;274(21):5659-68. Epub 2007 Oct 8. The hydroxyproline motif of male sex peptide elicits the innate immune response in Drosophila females. Domanitskaya EV, Liu H, Chen S, Kubli E. Zoologisches Institut Universität Zürich-Irchel, Zürich, Switzerland. Seminal fluid elicits a variety of physiological and behavioral changes in insect females. In Drosophila melanogaster females, sex peptide (SP) is the major seminal agent eliciting oviposition and reduction of receptivity. But SP also has many other effects; for example, it stimulates food intake, egg production, ovulation, juvenile hormone production and antimicrobial peptide synthesis. Thus, SP very probably has several receptors. To identify putative targets and signaling cascades, we studied the genome-wide regulation of genes by microarray analysis of RNA isolated from females after mating with wild-type males or males lacking SP, respectively. In addition, we studied the effects of SP on the proteome of females. Sex peptide regulates gene activity differentially in the head and in the abdomen. Genes coding for unspecific antimicrobial peptides are specifically transcribed in the abdomen, e.g. the antimicrobial peptide drosocin in epithelial tissues of the female genital tract (oviduct and calyx). Hence, SP elicits a systemic [Peng J, Zipperlen P & Kubli E (2005) Curr Biol15, 1690-1694] and an epithelial immune response. Ectopic expression of SP in the fat body of transgenic virgin females (with subsequent secretion into the hemolymph) does not elicit drosocin synthesis in the genital tract. Thus, the receptors for the stimulation of the systemic and the epithelial responses by SP are compartmentalized. The hydroxyproline (P*) motif of SP, P*TKFP*IP*SP*NP*, is identified as a novel elicitor of the innate immune response. We suggest that SP acts by chemical mimicry of sugar components of the bacterial cell wall. Thus, SP may induce the immune system via pattern recognition receptors. Publication Types: Research Support, Non-U.S. Gov't PMID: 17922838 [PubMed - indexed for MEDLINE] 347: Vet Res Commun. 2007 Aug;31 Suppl 1:385-8. Detection of genetically modified organisms in food: comparison among three different DNA extraction methods. Vodret B, Milia M, Orani MG, Serratrice G, Mancuso MR. Zooprofilattic Institute of Sardinia, Feed Hygiene Unit, Sassari, Italy. bruna.vodret@izs-sardegna.it PMID: 17682920 [PubMed - indexed for MEDLINE] 348: Nat Biotechnol. 2007 Oct;25(10):1065-6. Erratum in: Nat Biotechnol. 2008 Feb;26(2):241. Europe's anti-GM stance to presage animal feed shortage? Mitchell P. Publication Types: News PMID: 17921975 [PubMed - indexed for MEDLINE] 349: Food Chem Toxicol. 2007 Nov;45(11):2073-85. Epub 2007 Aug 30. Report of an Expert Panel on the reanalysis by of a 90-day study conducted by Monsanto in support of the safety of a genetically modified corn variety (MON 863). Doull J, Gaylor D, Greim HA, Lovell DP, Lynch B, Munro IC. Pharmacology, Toxicology and Therapeutics, Division of Toxicology, Department of Pharmacology, The University of Kansas Medical Center, 1018A Briedenthal Building, 3901 Rainbow Boulevard, Kansas City, KS 66160-7417, USA. MON 863, a genetically engineered corn variety that contains the gene for modified Bacillus thuringiensis Cry3Bb1 protein to protect against corn rootworm, was tested in a 90-day toxicity study as part of the process to gain regulatory approval. This study was reanalyzed by Séralini et al. who contended that the study showed possible hepatorenal effects of MON 863. An Expert Panel was convened to assess the original study results as analyzed by the Monsanto Company and the reanalysis conducted by Séralini et al. The Expert Panel concludes that the Séralini et al. reanalysis provided no evidence to indicate that MON 863 was associated with adverse effects in the 90-day rat study. In each case, statistical findings reported by both Monsanto and Séralini et al. were considered to be unrelated to treatment or of no biological or clinical importance because they failed to demonstrate a dose-response relationship, reproducibility over time, association with other relevant changes (e.g., histopathology), occurrence in both sexes, difference outside the normal range of variation, or biological plausibility with respect to cause-and-effect. The Séralini et al. reanalysis does not advance any new scientific data to indicate that MON 863 caused adverse effects in the 90-day rat study. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 17900781 [PubMed - indexed for MEDLINE] 350: Int J Food Microbiol. 2007 Oct 20;119(1-2):116-25. Epub 2007 Aug 19. Climatic models to predict occurrence of Fusarium toxins in wheat and maize. Schaafsma AW, Hooker DC. University of Guelph, Ridgetown Campus, Ridgetown, Ontario, Canada N0P 2C0. aschaafs@ridgetownc.uoguelph.ca Although forecasting Fusarium infections have useful implications, it may be argued that forecasting Fusarium toxins is more useful to help reduce their entry into the food chain. Several disease incidence models have been commercialized for wheat, but only one toxin prediction model from Ontario, Canada, "DONcast", has been validated extensively and commercialized to date for wheat, and another has been proposed for maize. In the development of these predictive tools, the variation in toxin levels associated with year and agronomic effects was estimated from simple linear models using wheat and maize samples taken from farm fields. In wheat, environment effects accounted for 48% of the variation in deoxynivalenol (DON) across all fields, followed by variety (27%), and previous crop (14 to 28%). In maize, hybrid accounted for 25% of the variation of either DON or fumonisin, followed by environment (12%), and when combined 42% of the variability was accounted for. The robust site-specific, DON forecast model accounted for up to 80% of the variation in DON, and has been used commercially for 5 years in Canada. Forecasting DON and fumonisins in maize is more difficult, because of its greater exposure to infection, the role of wounding in infection, the more important role of hybrid susceptibility, and the vast array of uncharacterized hybrids available in the marketplace. Nevertheless, using data collected from controlled experiments conducted in Argentina and the Philippines, a model was developed to predict fumonisin concentration using insect damage and weather variables, accounting for 82% of the variability of fumonisins. Using mycotoxins as a measure of disease outcome, as opposed to disease symptoms, offers a more robust prediction of mycotoxin risk, and it accounts for mycotoxin accumulation that occurs frequently in the absence of any change in Fusarium symptoms. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 17900733 [PubMed - indexed for MEDLINE] 351: Adv Food Nutr Res. 2007;53:161-98. Designer milk. Sabikhi L. Dairy Technology Division, National Dairy Research Institute, Karnal 132001, Haryana, India. Dairy biotechnology is fast gaining ground in the area of altering milk composition for processing and/or animal and human health by employing nutritional and genetic approaches. Modification of the primary structure of casein, alteration in the lipid profile, increased protein recovery, milk containing nutraceuticals, and replacement for infant formula offer several advantages in the area of processing. Less fat in milk, altered fatty acid profiles to include more healthy fatty acids such as CLA and omega-fats, improved amino acid profiles, more protein, less lactose, and absence of beta-lactoglobulin (beta-LG) are some opportunities of "designing" milk for human health benefits. Transgenic technology has also produced farm animals that secrete in their milk, human lactoferrin, lysozyme, and lipase so as to simulate human milk in terms of quality and quantity of these elements that are protective to infants. Cow milk allergenicity in children could be reduced by eliminating the beta-LG gene from bovines. Animals that produce milk containing therapeutic agents such as insulin, plasma proteins, drugs, and vaccines for human health have been genetically engineered. In order to cater to animal health, transgenic animals that express in their mammary glands, various components that work against mastitis have been generated. The ultimate acceptability of the "designer" products will depend on ethical issues such as animal welfare and safety, besides better health benefits and increased profitability of products manufactured by the novel techniques. Publication Types: Review PMID: 17900499 [PubMed - indexed for MEDLINE] 352: Plant Mol Biol. 2007 Nov;65(5):693-705. Epub 2007 Sep 25. Comparative genome organization reveals a single copy of CBF in the freezing tolerant crucifer Thlaspi arvense. Zhou N, Robinson SJ, Huebert T, Bate NJ, Parkin IA. Saskatoon Research Centre, Agriculture and Agri-Food Canada, 107 Science Place, Saskatoon, SK, Canada S7N-0X2. The weedy crucifer species Thlaspi arvense has the ability to acclimate to lower temperatures than Arabidopsis thaliana and the related crop species, Brassica napus. As a step towards understanding the genetic basis for this enhanced low temperature response, we isolated and sequenced 8.7 kb of genomic DNA encompassing the T. arvense CBF locus. CBF is a transcription factor believed to play a pivotal role in the development of plant freezing tolerance. Sequence analysis revealed that T. arvense contains a single copy of CBF, whereas the co-linear, homologous region in A. thaliana contains three tandem copies. Genes that flank CBF in A. thaliana are also present in a co-linear arrangement in T. arvense. Comparative sequence alignment also revealed the presence of conserved sequence blocks between T. arvense and A. thaliana promoter regions. The expression of T. arvense CBF responds rapidly to low temperature but not demonstrably to ABA, dehydration or high salt, which is comparable to that of the A. thaliana CBF genes. Over-expression of Ta-CBF in transgenic A. thaliana resulted in the development of constitutive freezing tolerance, comparable to that of cold acclimated A. thaliana. Publication Types: Comparative Study Research Support, Non-U.S. Gov't PMID: 17899397 [PubMed - indexed for MEDLINE] 353: Plant Cell Rep. 2008 Jan;27(1):105-15. Epub 2007 Sep 27. Constitutive overexpression of a stress-inducible small GTP-binding protein PgRab7 from Pennisetum glaucum enhances abiotic stress tolerance in transgenic tobacco. Agarwal PK, Agarwal P, Jain P, Jha B, Reddy MK, Sopory SK. International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Road, New Delhi, 110 067, India. pagarwal@csmcri.org The Rab GTPases are important components of endocytic network in plant cells. Endocytosis participates in the cell's reaction to extracellular stimuli by desensitizing, down-regulating or recycling receptors and membrane proteins. Rab7 is a small GTP-binding protein involved in intracellular vesicle trafficking from late endosome to the vacuole. We have isolated Rab7 cDNA from Pennisetum glaucum, a relatively drought-stress tolerant food grain crop grown commonly in India, during cDNA-subtractive hybridization of dehydration-stress treated plants. The PgRab7 ORF, encoding 207 aminoacids, was over-expressed in E. coli. The recombinant PgRab7 protein showed GTP-binding and GTPase activity. Transcript expression of PgRab7 gene was differentially up-regulated by different environmental stimuli such as cold, dehydration and NaCl and also by a plant hormone IAA. Overexpression of PgRab7 gene enhanced tolerance to NaCl and mannitol in transgenic tobacco. Transgenic plants also had increased alkaline phosphatase (ALP) activity. These results show that PgRab7 is a potential candidate gene for developing both salinity and dehydration tolerance in planta. Publication Types: Research Support, Non-U.S. Gov't PMID: 17899098 [PubMed - indexed for MEDLINE] 354: Food Chem Toxicol. 2007 Dec;45(12):2372-80. Epub 2007 Aug 23. Proteomic analysis of ovomucoid hypersensitivity in mice by two-dimensional difference gel electrophoresis (2D-DIGE). Hobson DJ, Rupa P, Diaz GJ, Zhang H, Yang M, Mine Y, Turner PV, Kirby GM. Department of Pathobiology, University of Guelph, Guelph, Ontario, Canada N1G 2W1. There is a need to develop reliable methods to assess the safety of genetically modified and other novel foods. The aim of this study was to identify protein biomarkers of food allergy in mice exposed to ovomucoid (OVM), a major food allergen found in chicken egg white. BALB/c mice were repeatedly sensitized by gavage with OVM and cholera toxin (CT) and control mice were exposed to a mixture of amino acids with CT. At the endpoint, all mice were challenged intraperitoneally with OVM and alum. Type-1 hypersensitivity was confirmed in OVM-sensitized mice by observation of clinical signs of anaphylaxis and elevated levels of plasma histamine, OVM-specific IgE and OVM-specific IgG by ELISA. Differential protein expression was assessed in albumin-depleted plasma as well as in mesenteric lymph node, liver, spleen, and ileum by two-dimensional difference gel electrophoresis (2D-DIGE). Differentially expressed proteins were identified by liquid chromatography with tandem mass spectrometry. Plasma proteins overexpressed in OVM-sensitized mice included haptoglobin (41-fold), serum amyloid A (19-fold) and peroxiredoxin-2 (1.9-fold). Further validation of these plasma proteins in other animal models of food allergy with different food allergens is required to assess their potential as candidate biomarkers for use in evaluating the allergenicity of novel foods. Publication Types: Evaluation Studies Research Support, Non-U.S. Gov't PMID: 17897766 [PubMed - indexed for MEDLINE] 355: Health Res Policy Syst. 2007 Sep 26;5:10. Biofortification in China: policy and practice. Campos-Bowers MH, Wittenmyer BF. Department of Health Management and Policy, School of Public Health, University of North Texas Health Science Center, 3500 Camp Bowie Boulevard, Fort Worth, Texas 76107, USA. mcampos@hsc.unt.edu. ABSTRACT: Micronutrient deficiency undernutrition, due to insufficient levels of vitamins and minerals in the diet, remains one of the most prevalent and preventable nutritional problems in the world today. Micronutrient undernutrition is the most common form of malnutrition. Compared to the 180 million children with protein-energy malnutrition, 3.5-5 billion persons are iron-deficient, and 140-250 million persons are vitamin A-deficient. Micronutrient deficiencies diminish physical, cognitive, and reproductive development. Undernutrition is both a cause and a result of poor human health and achievement.Middle-income nations, such as China, also suffer from micronutrient undernutrition's effects. In China's poor western provinces, despite supplementation and fortification efforts, stunting and underweight (symptoms of micronutrient undernutrition) remain common. In recent decades, nutritional adequacy, in terms of available food energy, improved immensely, as the government made food security a top priority. A potential next step for China could be to address specifically micronutrient undernutrition. The paper aims to provide a discussion of policy issues relevant to biofortification, if China were to consider the implementation of this intervention in its rural provinces.Traditional nutritional interventions currently employ four main strategies: dietary modification, supplementation, commercial fortification, and biofortification. Biofortification, a relatively new technique, involves selectively breeding staple plant varieties to increase specific nutrient levels in plant tissues. Biofortification has the potential to provide benefits to humans, plants, and livestock; nourish nutrient-depleted soils; and help increase crop yields per acre. Biofortification methods include selective breeding, reducing levels of anti-nutrients, and increasing levels of substances that promote nutrient absorption.If China were to implement biofortification programs, with help from government agencies and international organizations, several policy questions would need to be addressed. The paper discusses several policy questions that pertain to the relationship between biofortified and genetically modified crops, human health and safety concerns, labeling of biofortified crops for consumers, consumer rights, potential environmental impacts, intellectual property rights, seed disbursement, government investment, private-sector research, and additional agricultural and commercial regulations. Biofortification has the potential to help alleviate the suffering, death, disability, and failure to achieve full human potential that results from micronutrient undernutrition-related diseases. Publication Types: Editorial PMID: 17897456 [PubMed - in process] 356: J Dairy Sci. 2007 Oct;90(10):4718-23. Effects of corn silage derived from a genetically modified variety containing two transgenes on feed intake, milk production, and composition, and the absence of detectable transgenic deoxyribonucleic acid in milk in Holstein dairy cows. Calsamiglia S, Hernandez B, Hartnell GF, Phipps R. Dpto. Ciència Animal i dels Aliments, Universitat Autónoma de Barcelona, 08193-Bellaterra, Spain. Sergio.Calsamiglia@uab.es The objectives were to compare the chemical composition, nutritive value, feed intake, milk production and composition, and presence in milk of transgenic DNA and the encoded protein Cry1Ab when corn silages containing 2 transgenes (2GM: herbicide tolerance: mepsps and insect resistance: cry1Ab) were fed as part of a standard total mixed ration (TMR) compared with a near isogenic corn silage (C) to 8 multiparous lactating Holstein dairy cows in a single reversal design study. Cows were fed a TMR ration ad libitum and milked twice daily. Diets contained [dry matter (DM) basis] 45% corn silage, 10% alfalfa hay, and 45% concentrate (1.66 Mcal of net energy for lactation/kg of DM, 15.8% crude protein, 35% neutral detergent fiber, and 4.1% fat). Each period was 28-d long. During the last 4 d of each period, feed intake and milk production data were recorded and milk samples taken for compositional analysis, including the presence of transgenic DNA and Cry1Ab protein. There was no significant difference in the chemical composition between C and 2GM silages, and both were within the expected range (37.6% DM, 1.51 Mcal of net energy for lactation/kg, 8.6% crude protein, 40% neutral detergent fiber, 19.6% acid detergent fiber, pH 3.76, and 62% in vitro DM digestibility). Cows fed the 2GM silage produced milk with slightly higher protein (3.09 vs. 3.00%), lactose (4.83 vs. 4.72%) and solids-not-fat (8.60 vs. 8.40%) compared with C. However, the yield (kg/d) of milk (36.5), 3.5% fat-corrected milk (34.4), fat (1.151), protein (1.106), lactose (1.738), and solids-not-fat (3.094), somatic cell count (log10: 2.11), change in body weight (+7.8 kg), and condition score (+0.09) were not affected by type of silage, indicating no overall production difference. All milk samples were negative for the presence of transgenic DNA from either trait or the Cry1Ab protein. Results indicate that the 2GM silage modified with 2 transgenes did not affect nutrient composition of the silages and had no effect on animal performance and milk composition. No transgenic DNA and Cry1Ab protein were detected in milk. Publication Types: Randomized Controlled Trial PMID: 17881694 [PubMed - indexed for MEDLINE] 357: Pest Manag Sci. 2007 Nov;63(11):1107-15. Altered pesticide use on transgenic crops and the associated general impact from an environmental perspective. Kleter GA, Bhula R, Bodnaruk K, Carazo E, Felsot AS, Harris CA, Katayama A, Kuiper HA, Racke KD, Rubin B, Shevah Y, Stephenson GR, Tanaka K, Unsworth J, Wauchope RD, Wong SS. RIKILT-Institute of Food Safety, Wageningen University and Research Centre, Wageningen, Holland. gijs.kleter@wur.nl The large-scale commercial cultivation of transgenic crops has undergone a steady increase since their introduction 10 years ago. Most of these crops bear introduced traits that are of agronomic importance, such as herbicide or insect resistance. These traits are likely to impact upon the use of pesticides on these crops, as well as the pesticide market as a whole. Organizations like USDA-ERS and NCFAP monitor the changes in crop pest management associated with the adoption of transgenic crops. As part of an IUPAC project on this topic, recent data are reviewed regarding the alterations in pesticide use that have been observed in practice. Most results indicate a decrease in the amounts of active ingredients applied to transgenic crops compared with conventional crops. In addition, a generic environmental indicator -- the environmental impact quotient (EIQ) -- has been applied by these authors and others to estimate the environmental consequences of the altered pesticide use on transgenic crops. The results show that the predicted environmental impact decreases in transgenic crops. With the advent of new types of agronomic trait and crops that have been genetically modified, it is useful to take also their potential environmental impacts into account. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 17880042 [PubMed - indexed for MEDLINE] 358: Sci China C Life Sci. 2007 Oct;50(5):573-9. Progress in the evaluation of transgenic fish for possible ecological risk and its containment strategies. Hu W, Wang Y, Zhu Z. State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China. Genetically improved transgenic fish possess many beneficial economic traits; however, the commercial aquaculture of transgenic fish has not been performed till date. One of the major reasons for this is the possible ecological risk associated with the escape or release of the transgenic fish. Using a growth hormone transgenic fish with rapid growth characteristics as a subject, this paper analyzes the following: the essence of the potential ecological risks posed by transgenic fish; ecological risk in the current situation due to transgenic fish via one-factor phenotypic and fitness analysis, and mathematical model deduction. Then, it expounds new ideas and the latest findings using an artificially simulated ecosystem for the evaluation of the ecological risks posed by transgenic fish. Further, the study comments on the strategies and principles of controlling these ecological risks by using a triploid approach. Based on these results, we propose that ecological risk evaluation and prevention strategies are indispensable important components and should be accompanied with breeding research in order to provide enlightments for transgenic fish breeding, evaluation of the ecological risks posed by transgenic fish, and development of containment strategies against the risks. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 17879053 [PubMed - indexed for MEDLINE] 359: Med Pregl. 2007 Mar-Apr;60(3-4):195-7. [Diseases caused by bacteria and rickettsia in biological warfare and bioterrorism] [Article in Serbian] Bojić I, Vukadinov J, Minić S. Specijalisticka ordinacija Dr Bojić, Beograd. drbojic@net.yu INTRODUCTION: Until recently, the use of biological weapons was considered more from an academic than practical point of view. The list of agents and/or toxins that can be used as biological weapons is long. Some of them are highly lethal, while others cause morbidity and disability. BIOLOGICAL WEAPONS: Bacteria, rickettsia, viruses, fungi, protozoa and toxins can all be used as biological weapons. The infection may be acquired by inhalation of aerosols, ingestion of contaminated water or food or direct contact with infectious agents. Early recognition, diagnosis and treatment of these patients is of utmost importance. Special attention must be given to the use of genetically modified microorganisms. Medical protection from biological weapons is very important as well as continuous education. CONCLUSION: This article describes the main clinical characteristics of anthrax, cholera, plague, Q fever, tularemia, brucellosis, and glanders, as biological weapons, their diagnostics, treatment and basic prevention measures. Publication Types: English Abstract Review PMID: 17853736 [PubMed - indexed for MEDLINE] 360: Fortune. 2007 Jul 9;156(1):74-8, 80. Attack of the mutant rice. Gunther M. Publication Types: News PMID: 17853593 [PubMed - indexed for MEDLINE] 361: Transgenic Res. 2008 Aug;17(4):573-85. Epub 2007 Sep 13. Enhancing the carotenoid content of Brassica napus seeds by downregulating lycopene epsilon cyclase. Yu B, Lydiate DJ, Young LW, Schäfer UA, Hannoufa A. Agriculture and Agri-Food Canada, Saskatoon Research Centre, 107 Science Place, Saskatoon, SK, Canada. The accumulation of carotenoids in higher plants is regulated by the environment, tissue type and developmental stage. In Brassica napus leaves, beta-carotene and lutein were the main carotenoids present while petals primarily accumulated lutein and violaxanthin. Carotenoid accumulation in seeds was developmentally regulated with the highest levels detected at 35-40 days post anthesis. The carotenoid biosynthesis pathway branches after the formation of lycopene. One branch forms carotenoids with two beta rings such as beta-carotene, zeaxanthin and violaxanthin, while the other introduces both beta- and epsilon-rings in lycopene to form alpha-carotene and lutein. By reducing the expression of lycopene epsilon-cyclase (epsilon-CYC) using RNAi, we investigated altering carotenoid accumulation in seeds of B. napus. Transgenic seeds expressing this construct had increased levels of beta-carotene, zeaxanthin, violaxanthin and, unexpectedly, lutein. The higher total carotenoid content resulting from reduction of epsilon-CYC expression in seeds suggests that this gene is a rate-limiting step in the carotenoid biosynthesis pathway. epsilon-CYC activity and carotenoid production may also be related to fatty acid biosynthesis in seeds as transgenic seeds showed an overall decrease in total fatty acid content and minor changes in the proportions of various fatty acids. Publication Types: Research Support, Non-U.S. Gov't PMID: 17851775 [PubMed - indexed for MEDLINE] 362: Nat Biotechnol. 2007 Sep;25(9):981-7. Comment in: Nat Biotechnol. 2007 Dec;25(12):1351-4; author reply 1359-60. Nat Biotechnol. 2007 Dec;25(12):1354-5; author reply 1359-60. Nat Biotechnol. 2007 Dec;25(12):1355-6; author reply 1359-60. Nat Biotechnol. 2007 Dec;25(12):1355; author reply 1359-60. Nat Biotechnol. 2007 Dec;25(12):1355; author reply 1359-60. Nat Biotechnol. 2007 Dec;25(12):1356-8. Nat Biotechnol. 2007 Dec;25(12):1356; author reply 1359-60. GM soybeans and health safety--a controversy reexamined. Marshall A. PMID: 17846624 [PubMed - indexed for MEDLINE] 363: Philos Trans R Soc Lond B Biol Sci. 2008 Feb 27;363(1492):761-76. Biological control and sustainable food production. Bale JS, van Lenteren JC, Bigler F. School of Biosciences, University of Birmingham, Edgbaston, Birmingham B15 2TT, UK. j.s.bale@bham.ac.uk The use of biological control for the management of pest insects pre-dates the modern pesticide era. The first major successes in biological control occurred with exotic pests controlled by natural enemy species collected from the country or area of origin of the pest (classical control). Augmentative control has been successfully applied against a range of open-field and greenhouse pests, and conservation biological control schemes have been developed with indigenous predators and parasitoids. The cost-benefit ratio for classical biological control is highly favourable (1:250) and for augmentative control is similar to that of insecticides (1:2-1:5), with much lower development costs. Over the past 120 years, more than 5000 introductions of approximately 2000 non-native control agents have been made against arthropod pests in 196 countries or islands with remarkably few environmental problems. Biological control is a key component of a 'systems approach' to integrated pest management, to counteract insecticide-resistant pests, withdrawal of chemicals and minimize the usage of pesticides. Current studies indicate that genetically modified insect-resistant Bt crops may have no adverse effects on the activity or function of predators or parasitoids used in biological control. The introduction of rational approaches for the environmental risk assessment of non-native control agents is an essential step in the wider application of biological control, but future success is strongly dependent on a greater level of investment in research and development by governments and related organizations that are committed to a reduced reliance on chemical control. Publication Types: Review PMID: 17827110 [PubMed - indexed for MEDLINE] 364: Nature. 2007 Sep 6;449(7158):9. Biotech crop rules get rewrite. Marris E. Publication Types: News PMID: 17805261 [PubMed - indexed for MEDLINE] 365: Endocr Rev. 2007 Oct;28(6):664-84. Epub 2007 Sep 4. Neuropeptide y receptor selective ligands in the treatment of obesity. Kamiji MM, Inui A. Department of Gastroenterology, Faculty of Medicine, University of Sao Paulo, Ribeirão Preto Campus 14048-900, Ribeirão Preto-SP, Brazil. Obesity is a serious public health problem throughout the world, affecting both developed societies and developing countries. The central nervous system has developed a meticulously interconnected circuitry in order to keep us fed and in an adequate nutritional state. One of these consequences is that an energy-dense environment favors the development of obesity. Neuropeptide Y (NPY) is one of the most abundant and widely distributed peptides in the central nervous system of both rodents and humans and has been implicated in a variety of physiological actions. Within the hypothalamus, NPY plays an essential role in the control of food intake and body weight. Centrally administered NPY causes robust increases in food intake and body weight and, with chronic administration, can eventually produce obesity. NPY activates a population of at least six G protein-coupled Y receptors. NPY analogs exhibit varying degrees of affinity and specificity for these Y receptors. There has been renewed speculation that ligands for Y receptors may be of benefit for the treatment of obesity. This review highlights the therapeutic potential of Y(1), Y(2), Y(4), and Y(5) receptor agonists and antagonists as additional intervention to treat human obesity. Publication Types: Review PMID: 17785427 [PubMed - indexed for MEDLINE] 366: Plant Biotechnol J. 2008 Feb;6(2):135-45. Epub 2007 Sep 3. Alleviating peanut allergy using genetic engineering: the silencing of the immunodominant allergen Ara h 2 leads to its significant reduction and a decrease in peanut allergenicity. Dodo HW, Konan KN, Chen FC, Egnin M, Viquez OM. Department of Food and Animal Sciences, Food Biotechnology Laboratory, Alabama A&M University, Normal, AL 35762, USA. hortense.dodo@email.aamu.eduhortense.dodo@email.aamu.edu Peanut allergy is one of the most life-threatening food allergies and one of the serious challenges facing the peanut and food industries. Current proposed solutions focus primarily on ways to alter the immune system of patients allergic to peanut. However, with the advent of genetic engineering novel strategies can be proposed to solve the problem of peanut allergy from the source. The objectives of this study were to eliminate the immunodominant Ara h 2 protein from transgenic peanut using RNA interference (RNAi), and to evaluate the allergenicity of resulting transgenic peanut seeds. A 265-bp-long PCR product was generated from the coding region of Ara h 2 genomic DNA, and cloned as inverted repeats in pHANNIBAL, an RNAi-inducing plant transformation vector. The Ara h 2-specific RNAi transformation cassette was subcloned into a binary pART27 vector to construct plasmid pDK28. Transgenic peanuts were produced by infecting peanut hypocotyl explants with Agrobacterium tumefaciens EHA 105 harbouring the pDK28 construct. A total of 59 kanamycin-resistant peanut plants were regenerated with phenotype and growth rates comparable to wild type. PCR and Southern analyses revealed that 44% of plants stably integrated the transgene. Sandwich ELISA performed using Ara h 2-mAbs revealed a significant (P < 0.05) reduction in Ara h 2 content in several transgenic seeds. Western immunobloting performed with Ara h 2-mAb corroborated the results obtained with ELISA and showed absence of the Ara h 2 protein from crude extracts of several transgenic seeds of the T(0) plants. The allergenicity of transgenic peanut seeds expressed as IgE binding capacity was evaluated by ELISA using sera of patients allergic to peanut. The data showed a significant decrease in the IgE binding capacity of selected transgenic seeds compared to wild type, hence, demonstrating the feasibility of alleviating peanut allergy using the RNAi technology. PMID: 17784907 [PubMed - indexed for MEDLINE] 367: Sci Am. 2007 Sep;297(3):104-11. Sowing a gene revolution. Raney T, Pingali P. Agricultural and Development Economics Division, United Nations Food and Agriculture Organization, Rome. PMID: 17784631 [PubMed - indexed for MEDLINE] 368: J AOAC Int. 2007 Jul-Aug;90(4):1098-106. Characterization of genetically modified maize in weakly contaminated seed batches and identification of the origin of the adventitious contamination. Petit L, Pagny G, Baraige F, Nignol AC, Zhang D, Fach P. Agence Française de Sécurité Sanitaire des Aliments, Laboratoire d'Etudes et de Recherches sur la Qualité des Aliments et sur les Procédés Agro-Alimentaires, Unité EBA, 23 Avenue du Général De Gaulle, 94700 Maisons-Alfort, France. So far, relatively few genetically modified plants (GMPs) have been planted in the European Union (EU). However, in France, seed batches weakly contaminated by unidentified GM materials have recently been detected among commercial maize seeds (14 seed batches positive out of 447 analyzed). We have developed a 3-step approach to precisely identify the genetic modifications detected in such maize seed batches. First, to isolate GMPs derived from the contaminated seed batches, 10 000 maize seeds of each batch were planted and screened by polymerase chain reaction (PCR) on 100-plant batches, then on 10-plant subbatches, and finally, plant by plant. In a second step, specific identification of the individual GMPs was performed. Finally, to determine the origin of the contamination, each individual GMP was analyzed by simple sequence repeat (SSR) markers. The results showed that all batches were contaminated by few GM seeds, having a GM content < 0.1%. Finally, 12 individual GMPs have been isolated from 17 plant pools that were tested positive either for P35-S and/or T-Nos. MON810 and T25 transformation events approved for cultivation in the EU were detected in 7 individual GMPs. The other seed batches were contaminated by genetically modified organisms (GMOs) that are not approved in the EU, including GA21 or the stacking MON810/T25. Presumable identification of T14 was also achieved following sequencing of 1 individual GMP. The data also showed that most of the seed batches were contaminated by several transformation events. Finally, analysis of SSR markers indicated that the contaminations were essentially due to cross-pollination in the seed production process. Publication Types: Research Support, Non-U.S. Gov't PMID: 17784498 [PubMed - indexed for MEDLINE] 369: Plant Biotechnol J. 2007 Nov;5(6):791-801. Epub 2007 Aug 31. Stable expression of AtGA2ox1 in a low-input turfgrass (Paspalum notatum Flugge) reduces bioactive gibberellin levels and improves turf quality under field conditions. Agharkar M, Lomba P, Altpeter F, Zhang H, Kenworthy K, Lange T. Agronomy Department, Plant Molecular Biology Program, Genetics Institute, Institute of Food and Agricultural Sciences, University of Florida, Gainesville, FL 32611, USA. Bahiagrass (Paspalum notatum Flugge) is a prime candidate for molecular improvement of turf quality. Its persistence and low input characteristics made it the dominant utility turfgrass along highways in the south-eastern USA. However, the comparatively poor turf quality due to reduced turf density and prolific production of unsightly inflorescences currently limits the widespread use of bahiagrass as residential turf. Alteration of endogenous gibberellin (GA) levels by application of growth regulators or transgenic strategies has modified plant architecture in several crops. GA catabolizing AtGA2ox1 was subcloned under the control of the constitutive maize ubiquitin promoter and Nos 3'UTR. A minimal AtGA2ox1 expression cassette lacking vector backbone sequences was stably introduced into apomictic bahiagrass by biolistic gene transfer as confirmed by Southern blot analysis. Expression of AtGA2ox1 in bahiagrass as indicated by reverse transcription-polymerase chain reaction and Northern blot analysis resulted in a significant reduction of endogenous bioactive GA(1) levels compared to wild type. Interestingly, transgenic plants displayed an increased number of vegetative tillers which correlated with the level of AtGA2ox1 expression and enhanced turf density under field conditions. This indicates that GAs contribute to signalling the outgrowth of axillary buds in this perennial grass. Transgenic plants also showed decreased stem length and delayed flowering under controlled environment and field conditions. Consequently, turf quality following weekly mowing was improved in transgenic bahiagrass. Transgene expression and phenotype were transmitted to seed progeny. Argentine bahiagrass produces seeds asexually by apomixis, which reduces the risk of unintended transgene dispersal by pollen and results in uniform progeny. Publication Types: Research Support, Non-U.S. Gov't PMID: 17764521 [PubMed - indexed for MEDLINE] 370: Philos Trans R Soc Lond B Biol Sci. 2008 Feb 27;363(1492):905-13. The role of biotechnology for agricultural sustainability in Africa. Thomson JA. Department of Molecular and Cell Biology, University of Cape Town, Cape Town 7701, Republic of South Africa. jennifer.thomson@uct.ac.za Sub-Saharan Africa could have a shortfall of nearly 90Mt of cereals by the year 2025 if current agricultural practices are maintained. Biotechnology is one of the ways to improve agricultural production. Insect-resistant varieties of maize and cotton suitable for the subcontinent have been identified as already having a significant impact. Virus-resistant crops are under development. These include maize resistant to the African endemic maize streak virus and cassava resistant to African cassava mosaic virus. Parasitic weeds such as Striga attack the roots of crops such as maize, millet, sorghum and upland rice. Field trials in Kenya using a variety of maize resistant to a herbicide have proven very successful. Drought-tolerant crops are also under development as are improved varieties of local African crops such as bananas, cassava, sorghum and sweet potatoes. Publication Types: Review PMID: 17761472 [PubMed - indexed for MEDLINE] 371: J Virol. 2007 Nov;81(22):12285-97. Epub 2007 Aug 29. Recovery of Nicotiana benthamiana plants from a necrotic response induced by a nepovirus is associated with RNA silencing but not with reduced virus titer. Jovel J, Walker M, Sanfaçon H. Pacific Agri-Food Research Centre, Agriculture and Agri-Food Canada, Summerland, BC, Canada. Recovery of plants from virus-induced symptoms is often described as a consequence of RNA silencing, an antiviral defense mechanism. For example, recovery of Nicotiana clevelandii from a nepovirus (tomato black ring virus) is associated with a decreased viral RNA concentration and sequence-specific resistance to further virus infection. In this study, we have characterized the interaction of another nepovirus, tomato ringspot virus (ToRSV), with host defense responses during symptom induction and subsequent recovery. Early in infection, ToRSV induced a necrotic phenotype in Nicotiana benthamiana that showed characteristics typical of a hypersensitive response. RNA silencing was also activated during ToRSV infection, as evidenced by the presence of ToRSV-derived small interfering RNAs (siRNAs) that could direct degradation of ToRSV sequences introduced into sensor constructs. Surprisingly, disappearance of symptoms was not accompanied by a commensurate reduction in viral RNA levels. The stability of ToRSV RNA after recovery was also observed in N. clevelandii and Cucumis sativus and in N. benthamiana plants carrying a functional RNA-dependent RNA polymerase 1 ortholog from Medicago truncatula. In experiments with a reporter transgene (green fluorescent protein), ToRSV did not suppress the initiation or maintenance of transgene silencing, although the movement of the silencing signal was partially hindered. Our results demonstrate that although RNA silencing is active during recovery, reduction of virus titer is not required for the initiation of this phenotype. This scenario adds an unforeseen layer of complexity to the interaction of nepoviruses with the host RNA silencing machinery. The possibility that viral proteins, viral RNAs, and/or virus-derived siRNAs inactivate host defense responses is discussed. Publication Types: Research Support, Non-U.S. Gov't PMID: 17728227 [PubMed - indexed for MEDLINE] 372: Int J Food Microbiol. 2007 Oct 20;119(1-2):147-51. Epub 2007 Jul 31. Opportunities for biotechnology and policy regarding mycotoxin issues in international trade. Kendra DF, Dyer RB. USDA-Agricultural Research Service, National Center for Agricultural Utilization Research, 1815 N. University Street, Peoria, Illinois 61604, USA. david.kendra@ars.usda.gov Despite being introduced more than a decade ago, agricultural biotechnology still remains framed in controversy impacting both the global economy and international regulations. Controversies surrounding agricultural biotechnology produced crops and foods commonly focus on human and environmental safety, intellectual property rights, consumer choice, ethics, food security, poverty reduction and environmental conservation. Originally, some consumers were reluctant to accept the first generation agricultural biotechnology products because they appeared to primarily benefit agricultural producers; however, it is clear from continued evaluations that these technologies also improved both the safety and wholesomeness of food and helped improve the environment. Plants engineered to resist insect pests and tolerate less toxic pesticides resulted in improved yields thereby enabling farmers to produce more food per acre while reducing the need for herbicides, pesticides, and water and tilling. An indirect benefit of reduced pest damage in transgenic corn expressing genes to control insect pests is lower levels of mycotoxins, most notably those caused by the genus Fusarium. Mycotoxins are an important regulatory issue globally because of their toxic and carcinogenic potential to humans and animals. Complicating this issue is the fact that toxicological databases for mycotoxins are relatively incomplete compared to other food contaminants. Current debates about agricultural biotechnology and mycotoxins reveal significant differences in perception of associated risks and benefits. When faced with uncertainty, regulators tend to set limits as low as possible. Additionally, some regulators invoke the "Precautionary Principle" when limited information is available or disputes over interpretation exist for possible contaminants, including mycotoxins. A major concern regarding use of the "Precautionary Principle" is the appearance that regulators can justify setting any limit on the basis of inconclusive or unknown potential hazards of a contaminant which may significantly impact global trade because mycotoxin residues vary widely between countries. This paper describes the current economic and heath impact of these regulations and their impact on international trade. Publication Types: Review PMID: 17727996 [PubMed - indexed for MEDLINE] 373: Ir Med J. 2007 May;100(5):475-6. Genetically modified food and health--a cause for concern? Cullen E. Publication Types: Letter PMID: 17727126 [PubMed - indexed for MEDLINE] 374: Regul Toxicol Pharmacol. 2007 Oct;49(1):53-62. Comparative safety testing of genetically modified foods in a 90-day rat feeding study design allowing the distinction between primary and secondary effects of the new genetic event. Knudsen I, Poulsen M. Department of Toxicology and Risk Assessment, National Food Institute, Technical University of Denmark, 19 Moerkhoej Bygade, DK-2860 Soeborg, Denmark. This article discusses the wider experiences regarding the usefulness of the 90-day rat feeding study for the testing of whole foods from genetically modified (GM) plant based on data from a recent EU-project [Poulsen, M., Schrøder, M., Wilcks, A., Kroghsbo, S., Lindecrona, R.H., Miller, A., Frenzel, T., Danier, J., Rychlik, M., Shu, Q., Emami, K., Taylor, M., Gatehouse, A., Engel, K.-H., Knudsen, I., 2007a. Safety testing of GM-rice expressing PHA-E lectin using a new animal test design. Food Chem. Toxicol. 45, 364-377; Poulsen, M., Kroghsbo, S., Schrøder, M., Wilcks, A., Jacobsen, H., Miller, A., Frenzel, T., Danier, J., Rychlik, M., Shu, Q., Emami, K., Sudhakar, D., Gatehouse, A., Engel, K.-H., Knudsen, I., 2007b. A 90-day safety in Wistar rats fed genetically modified rice expressing snowdrop lectin Galanthus nivalis (GNA). Food Chem. Toxicol. 45, 350-363; Schrøder, M., Poulsen, M., Wilcks, A., Kroghsbo, S., Miller, A., Frenzel, T., Danier, J., Rychlik, M., Emami, K., Gatehouse, A., Shu, Q., Engel, K.-H., Knudsen, I., 2007. A 90-day safety study of genetically modified rice expressing Cry1Ab protein (Bacillus thuringiensis toxin) in Wistar rats. Food Chem. Toxicol. 45, 339-349]. The overall objective of the project has been to develop and validate the scientific methodology necessary for assessing the safety of foods from genetically modified plants in accordance with the present EU regulation. The safety assessment in the project is combining the results of the 90-day rat feeding study on the GM food with and without spiking with the pure novel gene product, with the knowledge about the identity of the genetic change, the compositional data of the GM food, the results from in-vitro/ex-vivo studies as well as the results from the preceding 28-day toxicity study with the novel gene product, before the hazard characterisation is concluded. The results demonstrated the ability of the 90-day rat feeding study to detect the biological/toxicological effects of the new gene product in the GM food. The authors consider on this basis that the 90-day, rodent feeding study with one high dose level and a dietary design based upon compositional data on the GM food and toxicity data on the gene product is sensitive and specific enough to verify the presence/absence of the biological/nutritional/toxicological effects of the novel gene insert and further by the use of spiking able to separate potentially unintended effects of the novel gene product from other unintended effects at the level of intake defined in the test and within the remit of the test. Recommendations for further work necessary in the field are given. Publication Types: Comparative Study Research Support, Non-U.S. Gov't PMID: 17719159 [PubMed - indexed for MEDLINE] 375: Int J Food Microbiol. 2007 Oct 20;119(1-2):126-30. Epub 2007 Jul 31. Strategies for managing Fusarium head blight and deoxynivalenol accumulation in wheat. Yuen GY, Schoneweis SD. Department of Plant Pathology, University of Nebraska-Lincoln, Lincoln, NE 68583-0722, United States. gyuen1@unl.edu Many mycotoxigenic fungi infect plant hosts and cause disease in the field. Therefore, control of field infection by these fungi is a critical step in managing mycotoxin accumulation in the harvested product. Fusarium graminearum, also known as Gibberella zeae, is the causal agent of Fusarium head blight (FHB), or scab, in cereals and is also the primary agent responsible for contamination of grain with deoxynivalenol (DON). Research efforts worldwide are devoted to the development of strategies to control field infection of wheat and barley by this pathogen. Strategies include the use of fungicides and biological control agents to protect flowering heads from infection. There is extensive effort in breeding for host resistance to infection and spread of the pathogen within the heads. Scientists are also seeking exogenous traits to introduce into cereals to enhance resistance. Cultural practices are also being examined, primarily as measures to reduce pathogen survival and inoculum production in crop residues. The successes and limitations of these strategies in the management of Fusarium head blight and deoxynivalenol are discussed. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 17716767 [PubMed - indexed for MEDLINE] 376: Mol Cell Neurosci. 2007 Oct;36(2):211-21. Epub 2007 Jul 24. The Drosophila ARC homolog regulates behavioral responses to starvation. Mattaliano MD, Montana ES, Parisky KM, Littleton JT, Griffith LC. Department of Biology and National Center for Behavior Genomics, Brandeis University, Waltham, MA 02454-9110, USA. The gene encoding dARC1, one of three Drosophila homologs of mammalian activity-regulated cytoskeleton-associated protein (ARC), is upregulated in both seizure and muscular hypercontraction mutants. In this study we generate a null mutant for dArc1 and show that this gene is not involved in synaptic plasticity at the larval neuromuscular junction or in formation or decay of short-term memory of courtship conditioning, but rather is a modifier of stress-induced behavior. dARC1 is expressed in a number of neurosecretory cells and mutants are starvation-resistant, exhibiting an increased time of survival in the absence of food. Starvation resistance is likely due to the fact that dArc1 mutants lack the normal hyperlocomotor response to starvation, which is almost universal in the animal kingdom. dARC1 acts in insulin-producing neurons of the pars intercerebralis to control this behavior, but does not appear to be a general regulator of insulin signaling. This suggests that there are multiple modes of communication between the pars and the ring gland that control starvation-induced behavioral responses. Publication Types: Research Support, N.I.H., Extramural Research Support, U.S. Gov't, Non-P.H.S. PMID: 17707655 [PubMed - indexed for MEDLINE] 377: Sci Eng Ethics. 2007 Mar;13(1):69-82. A case for a duty to feed the hungry: GM plants and the third world. Carter L. The School of History, Philosophy, Religion and Classics, The University of Queensland, Brisbane, Australia. l.carter@uq.edu.au This article is concerned with a discussion of the plausibility of the claim that GM technology has the potential to provide the hungry with sufficient food for subsistence. Following a brief outline of the potential applications of GM in this context, a history of the green revolution and its impact will be discussed in relation to the current developing world agriculture situation. Following a contemporary analysis of malnutrition, the claim that GM technology has the potential to provide the hungry with sufficient nourishment will be discussed within the domain of moral philosophy to determine whether there exists a moral obligation to pursue this end if and only if the technology proves to be relatively safe and effective. By using Peter Singer's duty of moral rescue, I argue that we have a moral duty to assist the third world through the distribution of such GM plants. I conclude the paper by demonstrating that my argument can be supported by applying a version of the Precautionary Principle on the grounds that doing nothing might be worse for the current situation. PMID: 17703610 [PubMed - indexed for MEDLINE] 378: Biotechnol J. 2007 Sep;2(9):1086-7. The difficulty of structuring and focusing the co-existence debate in Europe. Custers R. VIB, Gent, Belgium. Rene.Custers@vib.be The co-existence debate in Europe is wide and difficult. In this paper some recommendations are given on how to make progress in the debate. Not with the goal of pushing GMOs, but with the goal of achieving genuine freedom of choice. PMID: 17703493 [PubMed - indexed for MEDLINE] 379: Biotechnol J. 2007 Sep;2(9):1141-6. Transparent communication strategy on GMOs: will it change public opinion? Sinemus K, Egelhofer M. Genius Gmbh, Darmstadt, Germany. Kristina.Sinemus@genius.de Innovations are central for the economic growth; however, the use of new technologies needs to be widely accepted in the general public and the society as a whole. Biotechnology in general, and the use of genetic engineering in food production in particular are seen critically by the European public and perceived as "risky", and a transatlantic divide between European and US citizens has been observed. This review investigates the reasons for those differing perceptions and proposes new strategies to communicate the benefits of biotechnology in agriculture to a broader public. When analyzing the dialogue process that has taken place between public, scientists, governmental organizations and industry, questions arise on what has been done differently in Europe, in order to propose new, more successful and efficient communication strategies for the future. Publication Types: Review PMID: 17703492 [PubMed - indexed for MEDLINE] 380: Biotechnol J. 2007 Sep;2(9):1088-92. Intellectual property, genetically modified crops and bioethics. Adcock M. Department of Law, Durham University, Durham, UK. mike.adcock@durham.ac.uk The implementation of a new technology is almost always surrounded by a debate on the moral and social implications that may arise. The debate with regard to genetically modified (GM) crops has been one of the longest and most controversial. However, one area of the debate that receives less attention is the role that intellectual property can play. The introduction of an effective and yet appropriate intellectual property system addressing society's particular needs can eliminate some of these issues. This paper looks at whether the situation in Europe is meeting our current needs and also addresses the role intellectual property can play in the debate over the introduction of GM crops in developing countries. Publication Types: Review PMID: 17703487 [PubMed - indexed for MEDLINE] 381: Transgenic Res. 2007 Dec;16(6):675-88. Epub 2007 Aug 14. Genetically modified crops for the bioeconomy: meeting public and regulatory expectations. Chapotin SM, Wolt JD. US Agency for International Development, Washington, DC 20523, USA. As the United States moves toward a plant-based bioeconomy, a large research and development effort is focused on creating new feedstocks to meet biomass demand for biofuels, bioenergy, and specialized bioproducts, such as industrial compounds and biomaterial precursors. Most bioeconomy projections assume the widespread deployment of novel feedstocks developed through the use of modern molecular breeding techniques, but rarely consider the challenges involved with the use of genetically modified crops, which can include hurdles due to regulatory approvals, market adoption, and public acceptance. In this paper we consider the implications of various transgenic crops and traits under development for the bioeconomy that highlight these challenges. We believe that an awareness of the issues in crop and trait selection will allow developers to design crops with maximum stakeholder appeal and with the greatest potential for widespread adoption, while avoiding applications unlikely to meet regulatory approval or gain market and public acceptance. Publication Types: Review PMID: 17701080 [PubMed - indexed for MEDLINE] 382: Nature. 2007 Aug 16;448(7155):736. Geneticist trades plants for politics. Nina Fedoroff interviewed by Emma Marris. Fedoroff N. Publication Types: Interview PMID: 17700665 [PubMed - indexed for MEDLINE] 383: J Dairy Sci. 2007 Sep;90(9):4084-91. Use of human lysozyme transgenic goat milk in cheese making: effects on lactic acid bacteria performance. Scharfen EC, Mills DA, Maga EA. Department of Animal Science, University of California, Davis 95616, USA. Genetically engineered goats expressing elevated levels of the antimicrobial enzyme lysozyme in their milk were developed to improve udder health, product shelf life, and consumer well-being. The purpose of this study was to evaluate the effect of lysozyme on the development of lactic acid bacteria (LAB) throughout the cheese-making process. Raw and pasteurized milk from 7 lysozyme transgenic goats and 7 breed-, age-, and parity-matched nontransgenic controls was transformed into cheeses by using industry methods, and their microbiological load was evaluated. The numbers of colony-forming units of LAB were determined for raw and pasteurized goat milk, whey, and curd at d 2 and at d 6 or 7 of production. Selective plating media were used to enumerate lactococcal species separately from total LAB. Although differences in the mean number of colony-forming units between transgenic and control samples in raw milk, whey, and cheese curd were non-significant for both total LAB and lactococcal species from d 2 of production, a significant decrease was observed in both types of LAB among d 6 transgenic raw milk cheese samples. In pasteurized milk trials, a significant decrease in LAB was observed only in the raw milk of transgenic animals. These results indicate that lysozyme transgenic goat milk is not detrimental to LAB growth during the cheese-making process. PMID: 17699025 [PubMed - indexed for MEDLINE] 384: J Dairy Sci. 2007 Sep;90(9):4005-21. Invited review: Advances in starter cultures and cultured foods. Cogan TM, Beresford TP, Steele J, Broadbent J, Shah NP, Ustunol Z. Moorepark Food Reseach Centre, Teagasc, Fermoy, Ireland. With 2005 retail sales close to $4.8 million, cultured dairy products are driving the growth of dairy foods consumption. Starter cultures are of great industrial significance in that they play a vital role in the manufacturing, flavor, and texture development of fermented dairy foods. Furthermore, additional interest in starter bacteria has been generated because of the data accumulating on the potential health benefits of these organisms. Today, starter cultures for fermented foods are developed mainly by design rather than by the traditional screening methods and trial and error. Advances in genetics and molecular biology have provided opportunities for genomic studies of these economically significant organisms and engineering of cultures that focuses on rational improvement of the industrially useful strain. Furthermore, much research has been published on the health benefits associated with ingesting cultured dairy foods and probiotics, particularly their role in modulating immune function. The aim of this review is to describe some of the major scientific advances made in starter and non-starter lactic acid bacteria during the past 10 yr, including genomic studies on dairy starter cultures, engineering of culture attributes, advances in phage control, developments in methods to enumerate lactic acid bacteria and probiotics in dairy foods, and the potential role of cultured dairy foods in modulation of immune function. Publication Types: Review PMID: 17699017 [PubMed - indexed for MEDLINE] 385: Neuron. 2007 Aug 16;55(4):662-76. Dopamine mediates context-dependent modulation of sensory plasticity in C. elegans. Kindt KS, Quast KB, Giles AC, De S, Hendrey D, Nicastro I, Rankin CH, Schafer WR. Biomedical Sciences Graduate Program, University of California, San Diego, La Jolla CA 92093, USA. Dopamine has been implicated in the modulation of diverse forms of behavioral plasticity, including appetitive learning and addiction. An important challenge is to understand how dopamine's effects at the cellular level alter the properties of neural circuits to modify behavior. In the nematode C. elegans, dopamine modulates habituation of an escape reflex triggered by body touch. In the absence of food, animals habituate more rapidly than in the presence of food; this contextual information about food availability is provided by dopaminergic mechanosensory neurons that sense the presence of bacteria. We find that dopamine alters habituation kinetics by selectively modulating the touch responses of the anterior-body mechanoreceptors; this modulation involves a D1-like dopamine receptor, a Gq/PLC-beta signaling pathway, and calcium release within the touch neurons. Interestingly, the body touch mechanoreceptors can themselves excite the dopamine neurons, forming a positive feedback loop capable of integrating context and experience to modulate mechanosensory attention. Publication Types: Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't PMID: 17698017 [PubMed - indexed for MEDLINE] 386: Planta. 2007 Dec;227(1):199-209. Epub 2007 Aug 11. Specific role of LeMAN2 in the control of seed germination exposed by overexpression of the LeMAN3 gene in tomato plants. Belotserkovsky H, Berger Y, Shahar R, Wolf S. Faculty of Agricultural, Food and Environmental Quality Sciences, The Institute of Plant Sciences and Genetics in Agriculture and the Otto Warburg Minerva Center for Agricultural Biotechnology, The Hebrew University of Jerusalem, Rehovot, Israel. Endo-beta-mannanase is one of the key enzymes involved in the hydrolysis of the mannan-rich cell walls of tomato (Solanum lycopersicon) seeds. Two isoforms of endo-beta-mannanase have been characterized in tomato seeds: LeMAN2 is active in the micropylar area prior to germination and LeMAN1 is active after germination in all endosperm cells surrounding the cotyledons. To explore whether general mannanase activity in the endosperm cap is sufficient to promote germination, the gene encoding LeMAN3 was inserted into transgenic tomato plants under the control of a CaMV-35S promoter. Expression of LeMAN3 was evident in the endosperm cap and in the lateral endosperm of the transgenic seeds 10 min after imbibition. An activity test indicated increased activity of endo-beta-mannanase in the transgenic lines relative to the control line in all seed parts, during the first 20 h of imbibition. However, overexpression of LeMAN3 in transgenic seeds inhibited seed germination at both optimal and suboptimal temperatures. Detailed RT-PCR analyses revealed the transcription patterns of the genes encoding the various mannanase isoforms, and indicated a delay in LeMAN2 transcription in the endosperm cap of the transgenic seeds. Interestingly, tissue-print assays indicated similar mannanase activity in the micropylar areas for both transgenic and control seeds. These results indicate that overexpression of active endo-beta-mannanase in the endosperm cap is not sufficient to enable hydrolysis of the cell walls or to promote germination of tomato seeds. Cell-wall hydrolysis in these endosperm cells is under tight control and requires the specific activity of LeMAN2. Publication Types: Research Support, Non-U.S. Gov't PMID: 17694319 [PubMed - indexed for MEDLINE] 387: Food Chem Toxicol. 2007 Dec;45(12):2513-25. Epub 2007 Jun 21. History of safe use as applied to the safety assessment of novel foods and foods derived from genetically modified organisms. Constable A, Jonas D, Cockburn A, Davi A, Edwards G, Hepburn P, Herouet-Guicheney C, Knowles M, Moseley B, Oberdörfer R, Samuels F. Nestlé Research Centre, Vers-Chez-les-blanc 1000, Lausanne 26, Switzerland. Very few traditional foods that are consumed have been subjected to systematic toxicological and nutritional assessment, yet because of their long history and customary preparation and use and absence of evidence of harm, they are generally regarded as safe to eat. This 'history of safe use' of traditional foods forms the benchmark for the comparative safety assessment of novel foods, and of foods derived from genetically modified organisms. However, the concept is hard to define, since it relates to an existing body of information which describes the safety profile of a food, rather than a precise checklist of criteria. The term should be regarded as a working concept used to assist the safety assessment of a food product. Important factors in establishing a history of safe use include: the period over which the traditional food has been consumed; the way in which it has been prepared and used and at what intake levels; its composition and the results of animal studies and observations from human exposure. This paper is aimed to assist food safety professionals in the safety evaluation and regulation of novel foods and foods derived from genetically modified organisms, by describing the practical application and use of the concept of 'history of safe use'. Publication Types: Research Support, Non-U.S. Gov't PMID: 17692450 [PubMed - indexed for MEDLINE] 388: In Silico Biol. 2007;7(1):77-86. Improved prediction of allergenicity by combination of multiple sequence motifs. Kong W, Tan TS, Tham L, Choo KW. Bioinformatics Group, Nanyang Polytechnic, Singapore. KONG_Wai_Ming@nyp.gov.sg The identification and validation of protein allergens have become more important nowadays as more and more transgenic proteins are introduced into our food chains. Current allergen prediction algorithms focus on the identification of single motif or single allergen peptide for allergen detection. However, an analysis of the 575 allergen dataset shows that most allergens contain multiple motifs. Here, we present a novel algorithm that detects allergen by making use of combinations of motifs. Sensitivity of 0.772 and specificity of 0.904 were achieved by the proposed algorithm to predict allergen. The specificity of the proposed approach is found to be significantly higher than traditional single motif approaches. The high specificity of the proposed algorithm is useful in filtering out false positives, especially when laboratory resources are limited. PMID: 17688432 [PubMed - indexed for MEDLINE] 389: Transgenic Res. 2008 Aug;17(4):515-27. Epub 2007 Aug 9. Transgenic rice as a novel production system for Melanocarpus and Pycnoporus laccases. de Wilde C, Uzan E, Zhou Z, Kruus K, Andberg M, Buchert J, Record E, Asther M, Lomascolo A. CropDesign NV, a BASF Plant Science Company, Technologiepark 3, Zwijnaarde-Gent, Belgium. Laccases have numerous biotechnological applications, among them food processing. The widespread use of laccases has increased the demand for an inexpensive and safe source of recombinant enzyme. We explored the use of a rice-based system for the production of two fungal laccases derived from the ascomycete Melanocarpus albomyces and the basidiomycete Pycnoporus cinnabarinus. High-expression levels of active recombinant laccases were achieved by targeting expression to the endosperm of rice seeds. The laccase cDNAs were fused to a plant-derived signal sequence for targeting to the secretory pathway, and placed under the control of a constitutive seed-specific promoter fused to an intron for enhanced expression. This construct enabled the recovery of on average 0.1-1% of soluble laccase in total soluble proteins (TSP). The highest yields of recombinant laccases obtained in rice seeds were 13 and 39 ppm for riceMaL and ricePycL, respectively. The rice-produced laccases were purified and characterized. The wild-type and the recombinant proteins showed similar biochemical features in terms of molecular mass, pI, temperature and optimal pH and the N-terminus was correctly processed. Although presenting lower kinetic parameters, the rice-produced laccases were also suitable for the oxidative cross-linking of a food model substrate [maize-bran feruloylated arabinoxylans (AX)]. Publication Types: Research Support, Non-U.S. Gov't PMID: 17687629 [PubMed - indexed for MEDLINE] 390: Forum Nutr. 2007;60:183-95. Prospects for improving the nutritional quality of dairy and meat products. Coffey SG. CSIRO Livestock Industries, St. Lucia, Australia. s.coffey@irl.cri.nz Knowledge of the function of human and animal genes and their interactions is rapidly increasing as a result of the completion of sequencing efforts for the human, bovine and other genomes. Through transcriptomics, proteomics and metabolomics, we have the capacity to study the health effects of food compounds at the molecular level. The same tools that can assist the understanding of nutrigenomics in humans can also be applied to producing animal-derived foods with desired capacities to alter gene expression in humans. This, essentially, represents food taking another major step in value through the personalisation of health and nutrition. In its own right, nutrigenomics offers the potential to improve animal production enterprises through major health and productivity gains. Publication Types: Review PMID: 17684415 [PubMed - indexed for MEDLINE] 391: BMC Neurosci. 2007 Aug 6;8:65. Exploratory behaviour in NO-dependent cyclase mutants of Drosophila shows defects in coincident neuronal signalling. Tinette S, Zhang L, Garnier A, Engler G, Tares S, Robichon A. Université de Bourgogne, Dijon, France. tinette@cesg.cnrs.fr BACKGROUND: Drosophila flies explore the environment very efficiently in order to colonize it. They explore collectively, not individually, so that when a few land on a food spot, they attract the others by signs. This behaviour leads to aggregation of individuals and optimizes the screening of mates and egg-laying on the most favourable food spots. RESULTS: Flies perform cycles of exploration/aggregation depending on the resources of the environment. This behavioural ecology constitutes an excellent model for analyzing simultaneous processing of neurosensory information. We reasoned that the decision of flies to land somewhere in order to achieve aggregation is based on simultaneous integration of signals (visual, olfactory, acoustic) during their flight. On the basis of what flies do in nature, we designed laboratory tests to analyze the phenomenon of neuronal coincidence. We screened many mutants of genes involved in neuronal metabolism and the synaptic machinery. CONCLUSION: Mutants of NO-dependent cyclase show a specifically-marked behaviour phenotype, but on the other hand they are associated with moderate biochemical defects. We show that these mutants present errors in integrative and/or coincident processing of signals, which are not reducible to the functions of the peripheral sensory cells. Publication Types: Research Support, Non-U.S. Gov't PMID: 17683617 [PubMed - indexed for MEDLINE] 392: Transgenic Res. 2008 Aug;17(4):489-501. Epub 2007 Aug 8. Metabolic engineering of novel ketocarotenoid production in carrot plants. Jayaraj J, Devlin R, Punja Z. Department of Biological Sciences, Simon Fraser University, 8888 University Drive, Burnaby, British Columbia, Canada. Carotenoids constitute a vast group of pigments that are ubiquitous throughout nature. Carrot (Daucus carota L.) roots provide an important source of dietary beta-carotene (provitamin A), alpha-carotene and lutein. Ketocarotenoids, such as canthaxanthin and astaxanthin, are produced by some algae and cyanobacteria but are rare in plants. Ketocarotenoids are strong antioxidants that are chemically synthesized and used as dietary supplements and pigments in the aquaculture and neutraceutical industries. We engineered the ketocarotenoid biosynthetic pathway in carrot tissues by introducing a beta-carotene ketolase gene isolated from the alga Haematococcus pluvialis. Gene constructs were made with three promoters (double CaMV 35S, Arabidopsis-ubiquitin, and RolD from Agrobacterium rhizogenes). The pea Rubisco small sub-unit transit peptide was used to target the enzyme to plastids in leaf and root tissues. The phosphinothricin acetyl transferase (bar) gene was used as a selectable marker. Following Agrobacterium-mediated transformation, 150 plants were regenerated and grown in a glasshouse. All three promoters provided strong root expression, while the double CaMV 35S and Ubiquitin promoters also had strong leaf expression. The recombinant ketolase protein was successfully targeted to the chloroplasts and chromoplasts. Endogenous expression of carrot beta-carotene hydroxylases was up-regulated in transgenic leaves and roots, and up to 70% of total carotenoids was converted to novel ketocarotenoids, with accumulation up to 2,400 microg/g root dry weight. Astaxanthin, adonirubin, and canthaxanthin were most prevalent, followed by echinenone, adonixanthin and beta-cryptoxanthin. Our results show that carrots are suitable for biopharming ketocarotenoid production for applications to the functional food, neutraceutical and aquaculture industries. Publication Types: Research Support, Non-U.S. Gov't PMID: 17682834 [PubMed - indexed for MEDLINE] 393: Gastroenterology. 2007 Aug;133(2):517-28. Epub 2007 May 3. Comment in: Gastroenterology. 2007 Aug;133(2):706-9. Induction of ovalbumin-specific tolerance by oral administration of Lactococcus lactis secreting ovalbumin. Huibregtse IL, Snoeck V, de Creus A, Braat H, De Jong EC, Van Deventer SJ, Rottiers P. Center for Experimental and Molecular Medicine, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands. BACKGROUND AND AIMS: Obtaining antigen-specific immune suppression is an important goal in developing treatments of autoimmune, inflammatory, and allergic gastrointestinal diseases. Oral tolerance is a powerful means for inducing tolerance to a particular antigen, but implementing this strategy in humans has been difficult. Active delivery of recombinant autoantigens or allergens at the intestinal mucosa by genetically modified Lactococcus lactis (L lactis) provides a novel therapeutic approach for inducing tolerance. METHODS: We engineered the food grade bacterium L lactis to secrete ovalbumin (OVA) and evaluated its ability to induce OVA-specific tolerance in OVA T-cell receptor (TCR) transgenic mice (DO11.10). Tolerance induction was assessed by analysis of delayed-type hypersensitivity responses, measurement of cytokines and OVA-specific proliferation, phenotypic analysis, and adoptive transfer experiments. RESULTS: Intragastric administration of OVA-secreting L lactis led to active delivery of OVA at the mucosa and suppression of local and systemic OVA-specific T-cell responses in DO11.10 mice. This suppression was mediated by induction of CD4(+)CD25(-) regulatory T cells that function through a transforming growth factor beta-dependent mechanism. Restimulation of splenocytes and gut-associated lymph node tissue from these mice resulted in a significant OVA-specific decrease in interferon gamma and a significant increase in interleukin-10 production. Furthermore, Foxp3 and CTLA-4 were significantly up-regulated in the CD4(+)CD25(-) population. CONCLUSIONS: Mucosal antigen delivery by oral administration of genetically engineered L lactis leads to antigen-specific tolerance. This approach can be used to develop effective therapeutics for systemic and intestinal immune-mediated inflammatory diseases. Publication Types: Evaluation Studies Research Support, Non-U.S. Gov't PMID: 17681173 [PubMed - indexed for MEDLINE] 394: Duke Law J. 2007 Apr;56(6):1581-6. Beyond food and evil. Chen J. University of Louisville, Louis D. Brandeis School of Law. PMID: 17679180 [PubMed - indexed for MEDLINE] 395: Trends Biotechnol. 2007 Sep;25(9):376-84. Epub 2007 Jul 30. Humanizing infant milk formula to decrease postnatal HIV transmission. Blais DR, Altosaar I. Department of Biochemistry, Microbiology and Immunology, Faculty of Medicine, University of Ottawa, Ottawa, Ontario, K1H 8M5, Canada. There are currently no safe methods for feeding babies born from the 16 million HIV-infected women living in resource-constrained countries. Breast milk can transmit HIV, and formula feeding can lead to gastrointestinal illnesses owing to unsanitary conditions and the composition of milk formulations. There is therefore a need to ensure that breast milk substitutes provide optimal health outcomes. Given that the immune properties of several breast milk proteins are known, transgenic food crops could facilitate inexpensive and safe reconstitution of the beneficial breast milk proteome in infant formulae, while keeping the HIV virus at bay. At least seven breast milk immune proteins have already been produced in food crops, and dozens more proteins could potentially be produced if fortified formula proves effective in nursing newborns born to HIV-infected mothers. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 17659799 [PubMed - indexed for MEDLINE] 396: Neuroscience. 2007 Aug 24;148(2):371-4. Epub 2007 Jul 19. Co-regulation of cold-resistant food acquisition by insulin- and neuropeptide Y-like systems in Drosophila melanogaster. Lingo PR, Zhao Z, Shen P. Department of Cellular Biology, and Biomedical and Health Sciences Institute, 724 Biological Sciences Building, University of Georgia, Athens, GA 30602, USA. To survive, food-deprived animals may be forced to forage under hostile conditions. We attempt to use genetically tractable Drosophila melanogaster as a model to elucidate molecular and neural mechanisms that drive a forager to engage in risk-prone food acquisition. Here we describe a paradigm for assessing hunger-driven food acquisition by fly larvae at a deleteriously cold temperature. Genetic analyses reveal that the neural activity of NPFR1, a receptor of neuropeptide F (NPF, the sole fly homolog of neuropeptide Y or NPY), was required for cold-resistant feeding behavior of fasted larvae. Conversely, NPFR1 overexpression in fed larvae was sufficient to trigger cold-resistant feeding activity normally associated with fasted larvae. Furthermore, the fly insulin-like system, implicated in the transduction of hunger signals to the CNS, regulated negatively larval cold-resistant food acquisition. The results from this and our previous studies suggest that the fly NPY-like system is a central mediator of hunger-elicited resistance to diverse stressors that can be of thermal, gustatory or mechanical form. Publication Types: Research Support, N.I.H., Extramural PMID: 17658221 [PubMed - indexed for MEDLINE] 397: Food Nutr Bull. 2007 Jun;28(2 Suppl):S271-9. From harvest to health: challenges for developing biofortified staple foods and determining their impact on micronutrient status. Hotz C, McClafferty B. HarvestPlus in Washington, DC 20006-1002, USA. BACKGROUND: The use of conventional breeding techniques and biotechnology to improve the micronutrient quality of staple crops is a new strategy to address micronutrient deficiencies in developing countries. This strategy, referred to as "biofortification," is being developed and implemented through the international alliance of HarvestPlus to improve iron, zinc, and vitamin A status in low-income populations. OBJECTIVE: The objective of this paper is to review the challenges faced by nutritionists to determine and demonstrate the ability of biofortified crops to have an impact on the nutritional and health status of target populations. METHODS: We reviewed available published and unpublished information that is needed to design and evaluate this strategy, including issues related to micronutrient retention in staple foods, micronutrient bioavailability from plant foods, and evidence for the efficacy of high-micronutrient-content staple foods to improve micronutrient status. RESULTS: Further information is needed on the retention of micronutrients in staple foods, in particular of provitamin A carotenoids, when stored and prepared under different conditions. The low bioavailability of iron from staple foods and the ability to demonstrate an impact on zinc status are specific challenges that need to be addressed. In target countries, infections and other micronutrient deficiencies may confound the ability to affect micronutrient status, and this must be taken into account in community-based studies. CONCLUSIONS: Information to date suggests that biofortification has the potential to contribute to increased micronutrient intakes and improved micronutrient status. The success of this strategy will require the collaboration between health and agriculture sectors. Publication Types: Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. PMID: 17658073 [PubMed - indexed for MEDLINE] 398: Food Nutr Bull. 2007 Jun;28(2 Suppl):S258-70. Ensuring the supply of and creating demand for a biofortified crop with a visible trait: lessons learned from the introduction of orange-fleshed sweet potato in drought-prone areas of Mozambique. Low JW, Arimond M, Osman N, Cunguara B, Zano F, Tschirley D. Jan W Low was affiliated with the Department of Agricultural Economics, Michigan State University, East Lansing, Michigan, USA. j.low@cgiar.org BACKGROUND: Orange-fleshed sweet potato (OFSP) is a promising biofortified crop for sub-Saharan Africa because it has high levels of provitamin A carotenoids, the formed vitamin A is bioavailable, and white-fleshed sweet potato is already widely grown. OBJECTIVES: To examine whether farmers will adopt varieties with a distinct visible trait, young children will eat OFSP in sufficient quantities to improve vitamin A intake, OFSP can serve as an entry point for promoting a more diversified diet, and lessons can be drawn to assure sustained adoption. METHODS: The 2-year quasi-experimental intervention study followed households and children (n = 741; mean age, 13 months at baseline) through two agricultural cycles in drought prone-areas of Mozambique. RESULTS: OFSP is acceptable to farmers when introduced by using an integrated approach. In the second year, intervention children (n = 498) were more likely than control children (n = 243) to have consumed OFSP (54% vs. 4%), dark-green leaves (60% vs. 46%), or ripe papaya (65% vs. 42%) on 3 or more days in the previous week (p < .001 for all comparisons). Their vitamin A intakes were nearly eight times higher than those of control children (median, 426 vs. 56 1g RAE [retinol activity equivalents], p < .001). Diet diversification was limited by difficult agroecological conditions and low purchasing power. However, dietary diversity was higher among intervention than control children (32% vs. 9% consuming food from more than four groups; p < .001). CONCLUSIONS: An integrated OFSP-based approach had a positive impact on the vitamin A intake of young children. A market development component and improved vine multiplication systems are recommended to assure sustained adoption. Publication Types: Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. PMID: 17658072 [PubMed - indexed for MEDLINE] 399: Shokuhin Eiseigaku Zasshi. 2007 Jun;48(3):41-50. [A 52-week feeding study of genetically modified soybeans in F344 rats] [Article in Japanese] Sakamoto Y, Tada Y, Fukumori N, Tayama K, Ando H, Takahashi H, Kubo Y, Nagasawa A, Yano N, Yuzawa K, Ogata A, Kamimura H. Department of Environmental Health and Toxicology, Tokyo Metropolitan Institute of Public Health: 3-24-1 Hyakunin-cho, Shinjuku-ku, Tokyo 169-0073, Japan. A chronic feeding study to evaluate the safety of the genetically modified glyphosate-tolerant soybeans (GM soybeans) was conducted using rats. F344 DuCrj rats were fed diet containing GM soybeans or Non-GM soybeans at the concentration of 30% in basal diet. Non-GM soybeans were closely related strain of GM soybeans. These two diets were adjusted to an identical nutrient level. In this study, the influence of GM soybeans on rats was compared with that of the Non-GM soybeans, and furthermore, to assess the effect of soybeans themselves, the groups of rats fed GM and Non-GM soybeans were compared with a group fed commercial diet (CE-2). General conditions were observed daily and body weight and food consumption were recorded. At the intermediate examination (26 weeks), and at the termination (52 weeks), animals were subjected to hematology, serum biochemistry, and pathological examination. There were several differences in animal growth, food intake, serum biochemical parameters and histological findings between the rats fed the GM and/or Non-GM soybeans and the rats fed CE-2. However, body weight and food intake were similar for the rats fed the GM and Non-GM soybeans. Gross necropsy findings, hematological and serum biochemical parameters, organ weights, and pathological findings showed no meaningful difference between rats fed the GM and Non-GM soybeans. These results indicate that long-term intake of GM soybeans at the level of 30% in diet has no apparent adverse effect in rats. Publication Types: English Abstract PMID: 17657996 [PubMed - indexed for MEDLINE] 400: Philos Trans R Soc Lond B Biol Sci. 2008 Feb 12;363(1491):591-609. Genetic contributions to agricultural sustainability. Dennis ES, Ellis J, Green A, Llewellyn D, Morell M, Tabe L, Peacock WJ. CSIRO Plant Industry, GPO Box 1600, Canberra, Australian Capital Territory 2601, Australia. The current tools of enquiry into the structure and operation of the plant genome have provided us with an understanding of plant development and function far beyond the state of knowledge that we had previously. We know about key genetic controls repressing or stimulating the cascades of gene expression that move a plant through stages in its life cycle, facilitating the morphogenesis of vegetative and reproductive tissues and organs. The new technologies are enabling the identification of key gene activity responses to the range of biotic and abiotic challenges experienced by plants. In the past, plant breeders produced new varieties with changes in the phases of development, modifications of plant architecture and improved levels of tolerance and resistance to environmental and biotic challenges by identifying the required phenotypes in a few plants among the large numbers of plants in a breeding population. Now our increased knowledge and powerful gene sequence-based diagnostics provide plant breeders with more precise selection objectives and assays to operate in rationally planned crop improvement programmes. We can expect yield potential to increase and harvested product quality portfolios to better fit an increasing diversity of market requirements. The new genetics will connect agriculture to sectors beyond the food, feed and fibre industries; agri-business will contribute to public health and will provide high-value products to the pharmaceutical industry as well as to industries previously based on petroleum feedstocks and chemical modification processes. Publication Types: Review PMID: 17656342 [PubMed - indexed for MEDLINE] 401: Mol Biotechnol. 2007 Mar;35(3):215-23. Expression of a synthetic neutralizing epitope of porcine epidemic diarrhea virus fused with synthetic B subunit of Escherichia coli heat labile enterotoxin in rice endosperm. Oszvald M, Kang TJ, Tomoskozi S, Tamas C, Tamas L, Kim TG, Yang MS. Department of Biochemistry and Food Technology, Budapest University of Technology and Economics, Budapest, Hungary. Epitopes often require co-delivery with adjuvant and targeting proteins to enable recognition by the immune system, and this approach may also increase the efficacy of the antigen. In this study, we assess and describe the ability of transgenic rice plants to express a fusion protein consisting of the B-subunit of the Escherichia coli heat-labile enterotoxin (LTB) and a synthetic core-neutralizing epitope (COE) of porcine epidemic diarrhea virus (PEDV), inducing an enteric disease that is seen most predominantly in piglets. Both components of the fusion proteins were detected with Western blot analysis. The fusion protein was determined to assemble into pentamers, as was evidenced by its ability to bind to GM1 gangliosides, and evidenced an average level of expression in a transgenic rice endosperm. This indicates that the expression system of the plant is capable of generating a sizable amount of antigen, possibly allowing for the successful development of an edible vaccine. Publication Types: Research Support, Non-U.S. Gov't PMID: 17652785 [PubMed - indexed for MEDLINE] 402: Tsitol Genet. 2007 May-Jun;41(3):3-8; discussion 8-9. [International symposium "Biosafety issues in implementation of genetically modified organisms: new research approaches, regulation and public perception" and its appeal for support of agricultural biology development. May 10-14, 2006. Ialta, Ukraine] [Article in Russian] Blium IaB. Publication Types: Congresses PMID: 17649618 [PubMed - indexed for MEDLINE] 403: Plant Biotechnol J. 2007 Sep;5(5):555-69. Epub 2007 Jul 21. Improving containment strategies in biopharming. Murphy DJ. Biotechnology Unit, Division of Biological Sciences, University of Glamorgan, Treforest, CF37 1DL, UK. dmurphy2@glam.ac.uk This review examines the challenges of segregating biopharmed crops expressing pharmaceutical or veterinary agents from mainstream crops, particularly those destined for food or feed use. The strategy of using major food crops as production vehicles for the expression of pharmaceutical or veterinary agents is critically analysed in the light of several recent episodes of contamination of the human food chain by non-approved crop varieties. Commercially viable strategies to limit or avoid biopharming intrusion into the human food chain require the more rigorous segregation of food and non-food varieties of the same crop species via a range of either physical or biological methods. Even more secure segregation is possible by the use of non-food crops, non-crop plants or in vitro plant cultures as production platforms for biopharming. Such platforms already under development range from outdoor-grown Nicotiana spp. to glasshouse-grown Arabidopsis, lotus and moss. Amongst the more effective methods for biocontainment are the plastid expression of transgenes, inducible and transient expression systems, and physical containment of plants or cell cultures. In the current atmosphere of heightened concerns over food safety and biosecurity, the future of biopharming may be largely determined by the extent to which the sector is able to maintain public confidence via a more considered approach to containment and security of its plant production systems. Publication Types: Review PMID: 17645439 [PubMed - indexed for MEDLINE] 404: Environ Biosafety Res. 2006 Oct-Dec;5(4):237-8. Epub 2007 Jul 20. The Japanese experience with the Blue Book and subsequent activities in environmental biosafety of GM crops. Hayashi K. Society for Techno-Innovation of Agriculture, Forestry and Fisheries, Sankaido Bldg. 7F, 1-9-13 Akasaka, Minato-ku, Tokyo 107-0052, Japan. hayashi@staff.or.jp The Blue Book made a big contribution to the development of the Japanese Ministry of Agriculture, Forestry and Fisheries (MAFF) guidelines established in 1991 for almost all of the basic issues. However, the MAFF guidelines could not sufficiently cover some important areas that the Blue Book addressed well, such as potential consequences. This gap has been recovered substantially by a new law established in 2003. Japan still faces several important issues, including assessment of stacked products, potential consequences, comparative assessment, assessment of imported GM commodities and movement of concerned groups. PMID: 17640516 [PubMed - indexed for MEDLINE] 405: Environ Biosafety Res. 2006 Oct-Dec;5(4):227-31. Epub 2007 Jul 20. Perspective on OECD activities from a non-member country. Alexandrova N, Atanassov A. Agrobioinstitute, 8, Dragan Tzankov, Sofia 1164, Bulgaria. The OECD Blue Book, "Recombinant DNA: Safety Considerations" was published in 1986. The developed principles and concepts on the stepwise and case-by-case approach for risk assessment in the Blue Book have been used as a foundation for building national biosafety frameworks and international instruments for the regulation of the products of modern biotechnology. Twenty years after the Blue Book was published, OECD continues its activities on unique identifier systems, information-sharing, consensus documents for the biology of crops, trees and microorganisms with respect to harmonization of regulatory oversight and those of novel food and feed safety. These activities benefit, without any doubt, the international community at large, including OECD non-member countries. In order to strengthen its position in the international arena and to better respond to the needs of the changing world, OECD would be encouraged to participate in a more active manner in the technology transfer process and co-existence debate, together with continuing the organization's efforts on information-sharing and harmonization in the field of biotechnology and biosafety. PMID: 17640514 [PubMed - indexed for MEDLINE] 406: Environ Biosafety Res. 2006 Oct-Dec;5(4):223-5. Epub 2007 Jul 20. Current harmonization activities related to risk/safety assessment by the OECD Working Group on Harmonization of Regulatory Oversight in Biotechnology. MacDonald P, Yarrow S. Canadian Food Inspection Agency, 59 Camelot Drive, Ottawa, Ontario K1A 0Y9, Canada. pmacdonald@inspection.gc.ca PMID: 17640513 [PubMed - indexed for MEDLINE] 407: Environ Biosafety Res. 2006 Oct-Dec;5(4):219-22. Epub 2007 Jul 20. The impact of the OECD on the development of national/international risk/safety assessment frameworks. Gaugitsch H. Environmental Impact Assessment and Biosafety, Federal Environment Agency, Umweltbundesamt, Spittelauer Laende 5, A-1090 Vienna, Austria. helmut.gaugitsch@umweltbundesamt.at The role of OECD in the development of national and international risk/safety assessment frameworks is presented and discussed. The most relevant OECD bodies in this context have contributed a lot to the development of international biosafety frameworks, inter alia by organizing international conferences in the areas of food/feed and environmental safety of GMOs, focusing on practical and current scientific issues. The OECD Consensus Documents as well as the OECD Product Database have provided a good basis for risk/safety assessment frameworks and their implementation. The relevance for the OECD work in the international area is discussed, in particular, with respect to the work undertaken under the Cartagena Protocol on Biosafety, the Codex Alimentarius Commission and its work in biotechnology, as well as under the International Plant Protection Convention. An outlook and suggestions for future directions of OECD work in this area are presented. PMID: 17640512 [PubMed - indexed for MEDLINE] 408: Environ Biosafety Res. 2006 Oct-Dec;5(4):201-3. Epub 2007 Jul 20. 9th International Symposium on the Biosafety of Genetically Modified Organisms. Session VII: Risk management and monitoring. Schiemann J. Institute for Plant Virology, Microbiology and Biosafety, Federal Biological Research Centre for Agriculture and Forestry (BBA), Messeweg 11-12, 38104 Braunschweig, Germany. j.schiemann@bba.de Biosafety regulatory frameworks are intended to serve as mechanisms for ensuring the safe use of biotechnology products without imposing unacceptable risk to human health or the environment, or unintended constraints to technology transfer. In several regulatory systems GMO risk assessment has been separated from GMO risk management. As a consequence, risk assessment can be performed on a purely scientific basis, whereas risk management can take additional aspects (e.g. socio-economic or ethical) into consideration. For instance, the European Food Safety Authority (EFSA), the keystone of European Union risk assessment regarding food and feed safety, provides independent scientific advice and clear communication on existing and emerging risks in close collaboration with national authorities and in open consultation with its stakeholders. Risk management measures are not within the remit of EFSA, and remain the responsibility of the European Commission and Member States. Publication Types: Congresses PMID: 17640508 [PubMed - indexed for MEDLINE] 409: Mol Nutr Food Res. 2007 Aug;51(8):946-55. Serum testing of genetically modified soybeans with special emphasis on potential allergenicity of the heterologous protein CP4 EPSPS. Hoff M, Son DY, Gubesch M, Ahn K, Lee SI, Vieths S, Goodman RE, Ballmer-Weber BK, Bannon GA. Department of Allergology, Paul Ehrlich Institute, Langen, Germany. Roundup Ready soy contains the CP4-enolpyruvylshikimate-3-phosphate synthase (CP4 EPSPS) protein. Serum IgE from two distinct populations of soy-allergic patients were recruited to determine their IgE-binding specificity. One population consisted of 10 adult patients from Europe, whose primary diagnosis was soy food allergy with some also having mite allergy. In addition, 6 primarily mite-allergic, 6 food-allergic (celery, carrot, milk, shrimp, walnut, and apple), and 5 non-allergic patients were tested. Another population consisted of 13 children from Korea, whose primary diagnosis was atopic dermatitis and secondarily soy and egg sensitization. In addition, 11 non-allergic patients were tested. Each patient population was extensively characterized with respect to clinical symptoms, specific IgE (CAP) scores, and total IgE. Immunoblots and ELISA assays were developed using serum IgE from these patients and soy extracts, CP4 EPSPS, rice extract, ovalbumin, rubisco, purified major peanut allergen Ara h 2, the putative soy allergen Gly m Bd 30k and mite allergen Der f 2 proteins as the intended targets. Immunoblot results indicated that soy-allergic patients bound soy extracts but did not specifically bind rubisco or CP4 EPSPS. ELISA results were in general agreement with the immunoblot results except that rubisco bound significant quantities of serum IgE from some patients. These results indicate that the CP4 EPSPS protein does not bind significant quantities of IgE from two geographically distinct sensitive populations and there is no evidence for an increased allergenic potential of this biotech protein. PMID: 17639514 [PubMed - indexed for MEDLINE] 410: Plant Cell Rep. 2007 Nov;26(11):1999-2008. Epub 2007 Jul 18. LeERF1 positively modulated ethylene triple response on etiolated seedling, plant development and fruit ripening and softening in tomato. Li Y, Zhu B, Xu W, Zhu H, Chen A, Xie Y, Shao Y, Luo Y. Laboratory of Fruit Physiology and Molecular Biology, College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, People's Republic of China. To study the function of LeERF1 in ethylene triple response on etiolated seedling, plant development and fruit ripening and softening, LeERF1 gene was introduced into tomato (Lycopersicon esculentum cv. No. 4 Zhongshu) through Agrobacterium-mediated transformation. The sense LeERF1 and anti-sense LeERF1 transgenic tomato were obtained. Overexpression of LeERF1 in tomato caused the typical ethylene triple response on etiolated seedling. In the adult stage, 35S::LeERF1 resulted in morphological changes in the leaves of the LeERF1-sn lines. Anti-sense LeERF1 fruits had longer shelf life compared with wild-type tomato. The results of this manuscript indicated that LeERF1 positively mediated the ethylene signals, while the function of LeERF1 was verified for the first time to be positively related with ethylene triple response on etiolated seedling, plant development and fruit ripening and softening using LeERF1-sn, wt and LeERF1-as tomato. Publication Types: Research Support, Non-U.S. Gov't PMID: 17639404 [PubMed - indexed for MEDLINE] 411: Transgenic Res. 2008 Jun;17(3):393-402. Epub 2007 Jul 19. Impact of genetic structures on haploid genome-based quantification of genetically modified DNA: theoretical considerations, experimental data in MON 810 maize kernels (Zea mays L.) and some practical applications. Zhang D, Corlet A, Fouilloux S. GEVES Domaine du Magneraud, Laboratoire BioGEVES, B.P. 52, Surgeres 17700, France. david.zhang@geves.fr Real-time Polymerase Chain Reaction (PCR) based assays are widely used to estimate the content of genetically modified (GM) materials in food, feed and seed. It has been known that the genetic structures of the analyte can significantly influence the GM content expressed by the haploid genome (HG) % estimated using real-time PCR assays; this kind of influence is also understood as the impact of biological factors. The influence was first simulated at theoretical level using maize as a model. We then experimentally assessed the impact of biological factors on quantitative results, analysing by quantitative real-time PCR six maize MON 810 hybrid kernels with different genetic structures: (1) hemizygous from transgenic male parent, (2) hemizygous from transgenic female parent and (3) homozygous at the transgenic locus. The results obtained in the present study showed clear influences of biological factors on GM DNA quantification: 1% of GM materials by weight (wt) for the three genetic structures contained 0.39, 0.55 and 1.0% of GM DNA by HG respectively, from quantitative real-time PCR analyses. The relationships between GM wt% and GM HG% can be empirically established as: (1) in the case of the presence of a single GM trait: GM HG% = GM wt% x (0.5 +/- 0.167Y), where Y is the endosperm DNA content (%) in the total DNA of a maize kernel, (2) in the case of the presence of multiple GM traits: GM HG% = N x GM wt% x (0.5 +/- 0.167Y), where N is the number of GM traits (stacked or not) present in an unknown sample. This finding can be used by stakeholders related to GMO for empirical prediction from one unit of expression to another in the monitoring of seed and grain production chains. Practical equations have also been suggested for haploid copy number calculations, using hemizygous GM materials for calibration curves. Publication Types: Research Support, Non-U.S. Gov't PMID: 17638110 [PubMed - indexed for MEDLINE] 412: J Neurosci. 2007 Jul 18;27(29):7640-7. D1 dopamine receptor dDA1 is required in the mushroom body neurons for aversive and appetitive learning in Drosophila. Kim YC, Lee HG, Han KA. Department of Biology and The Huck Institute Neuroscience Graduate Program, Pennsylvania State University, University Park, Pennsylvania 16802, USA. Drosophila has robust behavioral plasticity to avoid or prefer the odor that predicts punishment or food reward, respectively. Both types of plasticity are mediated by the mushroom body (MB) neurons in the brain, in which various signaling molecules play crucial roles. However, important yet unresolved molecules are the receptors that initiate aversive or appetitive learning cascades in the MB. We have shown previously that D1 dopamine receptor dDA1 is highly enriched in the MB neuropil. Here, we demonstrate that dDA1 is a key receptor that mediates both aversive and appetitive learning in pavlovian olfactory conditioning. We identified two mutants, dumb1 and dumb2, with abnormal dDA1 expression. When trained with the same conditioned stimuli, both dumb alleles showed negligible learning in electric shock-mediated conditioning while they exhibited moderately impaired learning in sugar-mediated conditioning. These phenotypes were not attributable to anomalous sensory modalities of dumb mutants because their olfactory acuity, shock reactivity, and sugar preference were comparable to those of control lines. Remarkably, the dumb mutant's impaired performance in both paradigms was fully rescued by reinstating dDA1 expression in the same subset of MB neurons, indicating the critical roles of the MB dDA1 in aversive as well as appetitive learning. Previous studies using dopamine receptor antagonists implicate the involvement of D1/D5 receptors in various pavlovian conditioning tasks in mammals; however, these have not been supported by the studies of D1- or D5-deficient animals. The findings described here unambiguously clarify the critical roles of D1 dopamine receptor in aversive and appetitive pavlovian conditioning. Publication Types: Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. PMID: 17634358 [PubMed - indexed for MEDLINE] 413: Transgenic Res. 2007 Aug;16(4):467-78. Epub 2006 Nov 25. High level accumulation of alpha-glucan in maize kernels by expressing the gtfD gene from Streptococcus mutans. Zhang S, Dong JG, Wang T, Guo S, Glassman K, Ranch J, Nichols SE. Pioneer Hi-Bred International, Inc., a DuPont company, Johnston, IA 50131, USA. shirong.zhang@pioneer.com Glucosyltransferases (GTFs, EC.2.4.1.5) are bacterial enzymes that catalyze the polymerization of glucose residues from sucrose, leading to the production of high molecular weight glucan with alpha-1,3 /alpha-1,6 linkages. Such glucans, with many potential food and industrial applications, do not normally exist in higher plants. We fused a mutant form of the gtfD gene from Sreptococcus mutans with the maize (Zea mays L.) chloroplastic Brittle 1 transit peptide for amyloplast targeting. This construct, driven by the ubiquitin promoter, was introduced into maize by Agrobacterium-mediated transformation. We developed a novel HPLC-based method that enabled us differentially to distinguish transgene glucan from other endogenous polysaccharides in maize kernels. Using this method, we screened over 100 transgenic plants for the presence of GTF-produced glucan whose content varied between 0.8 and 14% of dry weight in the mature transgenic seeds. The mature transgenic plants were indistinguishable from wildtype plants in growth rate and morphology. Furthermore, starch granule size in the transgenic maize kernel was unaffected by the accumulation of the foreign polysaccharide. Mutation in Sh2, which encodes a subunit of ADP-glucose pyrophosphorylase, had no effect on glucan accumulation caused by gtfD expression. Our results indicated that high levels of novel carbohydrate polymer can be accumulated in crop plants through transgene technology. PMID: 17624807 [PubMed - indexed for MEDLINE] 414: Theor Appl Genet. 2007 Aug;115(4):549-60. Epub 2007 Jul 11. Investigation of rice transgene flow in compass sectors by using male sterile line as a pollen detector. Yuan QH, Shi L, Wang F, Cao B, Qian Q, Lei XM, Liao YL, Liu WG, Cheng L, Jia SR. College of Life Science and Agriculture, MOE Key Lab of Tropic Biological Resources, Hainan University, Haikou, 570228, China. Rice is the most important staple food in the world. The rapid development of transgenic rice and its future commercialization have raised concerns regarding transgene flow and its potential environmental risk. It is known that rice is a self-pollinated crop; the outcrossing rate between common cultivars is generally less than 1%. In order to improve the detection sensitivity of rice transgene flow, a male sterile (ms) line BoA with a high outcrossing rate was used as a pollen detector in this study. A concentric circle design was adopted, in which the transgenic rice B2 containing bar gene as a pollen donor was planted in the center circle and the recipient BoA was planted in eight compass sectors. The frequency of transgene flow in compass sectors was analyzed by continuous sampling to generate cumulative data. The results of two years with sound reproducibility demonstrated that the rice gene flow was closely associated with the wind direction. According to the mean frequency of transgene flow, the eight sectors can be divided into two groups: a higher frequency group downstream of the prevailing wind (DPW) with a mean frequency ranging from 6.47 to 26.24%, and a lower frequency group lateral to or upstream of the prevailing wind (UPW) with a mean frequency of 0.39 to 3.03%. On the basis of the cumulative data, 90-96% of the cumulative gene flow events occurred in the four DPW sectors, while it was 4-10% in the four UPW sectors. By using these systematic data, simulation models and isograms of transgene flow in the eight compass sectors were calculated and drawn, respectively. Publication Types: Research Support, Non-U.S. Gov't PMID: 17622509 [PubMed - indexed for MEDLINE] 415: Nat Biotechnol. 2007 Jul;25(7):717-8. The status of GM rice R&D in China. Wang Y, Johnston S. Publication Types: Letter PMID: 17621287 [PubMed - indexed for MEDLINE] 416: Curr Opin Allergy Clin Immunol. 2007 Aug;7(4):355-9. Animal models of anaphylaxis. Nauta A, Knippels L, Garssen J, Redegeld F. Numico Research, Wageningen, The Netherlands. PURPOSE OF REVIEW: In this review we will focus on recent advances in the role of mast cells in the pathophysiology of insect allergy and the possible mechanisms of mast cell activation in anaphylaxis. RECENT FINDINGS: Anaphylactic reactions in the mouse can be induced by several independent pathways involving immunoglobulin E, immunoglobulin free light chains, or immunoglobulin G. There is considerable evidence that mast cells play a central role in anaphylactic reactions to insect stings. Mast cells can be directly activated by components of insect venom or after allergic sensitization. Of interest is the observation that mast cells are not only effector cells in insect allergy, but may also play a protective role in preventing the development of severe anaphylactic responses or by controlling inflammatory reactions by modulation of antigen-specific T-cell responses. SUMMARY: The contribution of mast cells in anaphylactic responses to insect venom may be heterogeneous. On the one hand, activation of mast cells contributes to the pathology by the release of bioactive and tissue-damaging mediators. However, mast cell activation may neutralize constituents in insect venom and defend against the adverse effects of these toxins or they may modulate inflammation through downregulation of antigen-specific immune responses. Publication Types: Review PMID: 17620830 [PubMed - indexed for MEDLINE] 417: ScientificWorldJournal. 2007 Jun 22;7:1047-62. Cloning, structural characterization, and phylogenetic analysis of flower MADS-box genes from crocus (Crocus sativus L.). Tsaftaris AS, Polidoros AN, Pasentsis K, Kalivas A. Institute of Agrobiotechnology, Center for Research and Technology Hellas, Thermi, Greece. tsaft@certh.gr Crocus (Crocus sativus L.) is a crop species cultivated for its flowers and, more specifically, for its red stigmas. The flower of crocus is bisexual and sterile, since crocus is a triploid species. Its perianth consists of six petaloid tepals: three tepals in whorl 1 (outer tepals) and three tepals in whorl 2 (inner tepals). The androecium consists of three distinct stamens and the gynoecium consists of a single compound pistil with three carpels, a single three-branched style, and an inferior ovary. The dry form of the stigmas constitutes the commercial saffron used as a food additive, in the coloring industry, and in medicine. In order to uncover and understand the molecular mechanisms controlling flower development in cultivated crocus and its relative wild progenitor species, and characterize a number of crocus flower mutants, we have cloned and characterized different, full-length, cDNA sequences encoding MADS-box transcription factor proteins involved in flower formation. Here we review the different methods followed or developed for obtaining these sequences involving conventional 5 inverted exclamation markä 3 inverted exclamation markä RACE, as well as newly developed methods from our group, named Rolling Circle Amplification C RACE (RCA-RACE) and its modification named familyRCA-RACE (famRCA-RACE). Furthermore, the characteristics of the protein structure and their common and specific domains for each type of MADS-box transcription factors in this lower nongrass monocot belonging to the Iridaceae family are described. Finally, a phylogenetic tree of all the MADS-box sequences available in our lab is presented and discussed in relation to other data from studies of species of the Iridaceae group and closely related families from an evolutionary perspective. The structural and phylogenetic analyses are based on both published and unpublished data. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 17619787 [PubMed - indexed for MEDLINE] 418: Transgenic Res. 2007 Oct;16(5):541-55. Epub 2007 Jul 6. A multidisciplinary approach directed towards the commercial release of transgenic herbicide-tolerant rice in Costa Rica. Espinoza-Esquivel AM, Arrieta-Espinoza G. Centro de Investigación en Biología Celular y Molecular (CIBCM), Ciudad de la Investigación, Universidad de Costa Rica, San Jose, Costa Rica. amespino@racsa.co.cr This review discusses a multidisciplinary and multicomponent approach leading to the development and commercial release of transgenic Costa Rican rice varieties tolerant to the herbicide gluphosinate ammonium. We describe the field evaluations of the transgenic lines and their potential environmental impact, focusing on gene flow, particularly in relation to native wild Oryza species and weedy rice, based on trials performed in compliance with the national regulatory requirements of the country. We also present a socio-economic analysis of rice production in Costa Rica and the economic benefits of genetically modified (GM) rice as well as an environmental risk-benefit analysis for the deployment of GM rice. Additionally, food safety evaluation, intellectual property management, requirements for deregulation, and options for the commercialization of the new varieties are discussed. We also present results from a national survey aimed at assessing the level of support for GM crops in Costa Rica as this forms an integral component of our approach. Taken together, our results demonstrate that the adoption of these genetically improved rice varieties will provide clear benefits to Costa Rican rice growers and consumers. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 17619158 [PubMed - indexed for MEDLINE] 419: Transgenic Res. 2007 Oct;16(5):581-5. Epub 2007 Jul 6. Metabolic engineering of carotenoid accumulation by creating a metabolic sink. Li L, Van Eck J. USDA-ARS, Plant, Soil and Nutrition Laboratory, Cornell University, Ithaca, NY 14853, USA. ll37@cornell.edu Carotenoids are highly beneficial for human nutrition and health because they provide essential nutrients and important antioxidants in our diets. However, many food crops, especially the major staple crops contain only trace to low amounts of carotenoids. Although significant progress has been made in developing food crops rich in carotenoids by altering the expression of carotenoid biosynthetic genes, in many cases it has proved to be difficult to reach the desired levels of carotenoid enrichment. The recent identification and characterization of a novel gene mutation in cauliflower reveals that creating a metabolic sink to sequester carotenoids is an important mechanism to control carotenoid accumulation in plants. The successful demonstration of increased carotenoid accumulation in association with the formation of sink structures in transgenic crops offers a new and alternative approach to increase carotenoid content. Manipulation of the formation of metabolic sink along with the catalytic activity of the pathway may represent a promising strategy for maximally improving the nutritional quality of food crops. Publication Types: Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Review PMID: 17619157 [PubMed - indexed for MEDLINE] 420: Toxicology. 2007 Aug 16;238(1):1-14. Epub 2007 May 24. Erratum in: Toxicology. 2008 Jan 14;243(1-2):246. Mitra, Kalyan [added]. Adverse effect of organophosphate compounds, dichlorvos and chlorpyrifos in the reproductive tissues of transgenic Drosophila melanogaster: 70kDa heat shock protein as a marker of cellular damage. Gupta SC, Siddique HR, Mathur N, Mishra RK, Mitra K, Saxena DK, Chowdhuri DK. Embryotoxicology Section, Industrial Toxicology Research Centre, Lucknow 226001, India. The study highlights the adverse effects of organophosphate compounds dichlorvos and chlorpyrifos on reproduction in Drosophila. Freshly eclosed first instar larvae of Drosophila melanogaster transgenic for hsp70 (hsp70-lacZ) Bg(9) were fed on 0.015-150.0ppb dichlorvos and chlorpyrifos mixed food. Virgin flies eclosing from the normal and contaminated food were pair-mated to examine the effect of the test chemicals on reproduction of the exposed organisms. Expression of hsp70, sex peptide (SP or Acp70A), accessory gland protein (Acp36DE) and tissue damage was examined in reproductive organs of adult fly. Exposed organisms exhibited a dose-dependent significantly reduced reproductive outcome and males were found to be more sensitive than females. Hsp70 expression was restricted only within the testis lobes of male fly while it was not induced in the ovary of the female. In concurrence with absence of hsp70 expression in the accessory glands of male fly, tissue damage was evident in them. Acp70A and Acp36DE expression were found to be significantly downregulated at the higher concentrations of the test chemicals. The study suggests that (i) dichlorvos is more deleterious to fly reproduction compared to chlorpyrifos with an adverse effect on Acp70A and Acp36DE expression required to facilitate normal reproduction; (ii) hsp70 may be used as a marker of cellular damage against dichlorvos and chlorpyrifos in Drosophila. Publication Types: Research Support, Non-U.S. Gov't PMID: 17618723 [PubMed - indexed for MEDLINE] 421: Gen Comp Endocrinol. 2007 Oct-Dec;154(1-3):128-36. Epub 2007 Jun 3. Circulating corticosterone levels in breeding blue tits Parus caeruleus differ between island and mainland populations and between habitats. Müller C, Jenni-Eiermann S, Blondel J, Perret P, Caro SP, Lambrechts MM, Jenni L. Swiss Ornithological Institute, Luzernerstrasse 6, CH-6204 Sempach, Switzerland. claudia.mueller@vogelwarte.ch Little is known about whether adaptations to an insular life also involve adaptations in basal corticosterone levels or in the adrenocortical stress response, thus being part of a genetically based island syndrome. However, differences in corticosterone between island and mainland may also be a direct phenotypic response to differences in environmental conditions or may depend on individual characteristics of the animal such as body condition or parental investment. In this paper, we investigated whether insular (Island of Corsica) and mainland (nearby Southern France) blue tits Parus caeruleus populations differed in baseline and handling-stress induced corticosterone levels during the breeding season as a response to biological changes of insular biota. We also examined whether corticosterone levels of both mainland and insular blue tits differed between birds living in two different habitats (summergreen and evergreen oak woods) that differ in food availability and whether individual characteristics affected corticosterone levels. We found (a) differences in baseline corticosterone plasma levels between Corsica and the mainland, independent of regional differences in fat scores, (b) a regional difference in the relationship between corticosterone levels and brood size, (c) a difference in the rapidity of onset of the stress response to handling between habitats, independent of region, and (d) a negative relationship between body fat stores and baseline corticosterone levels independent of region. Reduced baseline corticosterone levels on Corsica may be a component of the insular syndrome, allowing birds to be less aggressive and to enhance parental investment despite higher breeding densities. We suggest that baseline corticosterone levels are only elevated if food availability affects directly the parents. However, when conditions deteriorate unexpectedly (as mimicked by handling stress), food allocation between parents and offspring needs to be re-adjusted in favor of the parents, possibly by increased circulating corticosterone levels. The switch to self-maintenance seems to be modified by the amount of body energy stores. Publication Types: Comparative Study PMID: 17617413 [PubMed - indexed for MEDLINE] 422: Plant Mol Biol. 2007 Sep;65(1-2):93-106. Epub 2007 Jul 5. Leaf rust resistance gene Lr1, isolated from bread wheat (Triticum aestivum L.) is a member of the large psr567 gene family. Cloutier S, McCallum BD, Loutre C, Banks TW, Wicker T, Feuillet C, Keller B, Jordan MC. Cereal Research Centre, Agriculture and Agri-Food Canada, R3T 2M9, Winnipeg, MB, Canada. scloutier@agr.gc.ca In hexaploid wheat, leaf rust resistance gene Lr1 is located at the distal end of the long arm of chromosome 5D. To clone this gene, an F(1)-derived doubled haploid population and a recombinant inbred line population from a cross between the susceptible cultivar AC Karma and the resistant line 87E03-S2B1 were phenotyped for resistance to Puccinia triticina race 1-1 BBB that carries the avirulence gene Avr1. A high-resolution genetic map of the Lr1 locus was constructed using microsatellite, resistance gene analog (RGA), BAC end (BE), and low pass (LP) markers. A physical map of the locus was constructed by screening a hexaploid wheat BAC library from cultivar Glenlea that is known to have Lr1. The locus comprised three RGAs from a gene family related to RFLP marker Xpsr567. Markers specific to each paralog were developed. Lr1 segregated with RGA567-5 while recombinants were observed for the other two RGAs. Transformation of the susceptible cultivar Fielder with RGA567-5 demonstrated that it corresponds to the Lr1 resistance gene. In addition, the candidate gene was also confirmed by virus-induced gene silencing. Twenty T (1) lines from resistant transgenic line T (0)-938 segregated for resistance, partial resistance and susceptibility to Avr1 corresponding to a 1:2:1 ratio for a single hemizygous insertion. Transgene presence and expression correlated with the phenotype. The resistance phenotype expressed by Lr1 seemed therefore to be dependant on the zygosity status. T (3)-938 sister lines with and without the transgene were further tested with 16 virulent and avirulent rust isolates. Rust reactions were all as expected for Lr1 thereby providing additional evidence toward the Lr1 identity of RGA567-5. Sequence analysis of Lr1 indicated that it is not related to the previously isolated Lr10 and Lr21 genes and unlike these genes, it is part of a large gene family. Publication Types: Research Support, Non-U.S. Gov't PMID: 17611798 [PubMed - indexed for MEDLINE] 423: J Agric Food Chem. 2007 Jul 25;55(15):5942-7. Epub 2007 Jul 4. Indicated detection of two unapproved transgenic rice lines contaminating vermicelli products. Akiyama H, Sasaki N, Sakata K, Ohmori K, Toyota A, Kikuchi Y, Watanabe T, Furui S, Kitta K, Maitani T. National Institute of Health Sciences, 1-18-1 Kamiyoga, Setagaya-ku, Tokyo 158-8501, Japan, Tokyo University of Agriculture and Technology, 2-24-16, Naka-cho, Koganei, Tokyo 184-8588, Japan. akiyama@nihs.go.jp We analyzed the DNA fragments extracted from four rice vermicelli products. The Bacillus thuringiensis (Bt) rice line, which has a construct similar to the GM Shanyou 63 line, was detected in some vermicelli products by identification of the junction region sequence between rice Act1 promoter and the Cry1Ac gene, and that between Cry1Ac and nos. In addition, we also detected a different Bt rice line by means of the junction region sequence between the maize ubiquitin promoter and cry1Ab gene and that between the cauliflower mosaic virus 35S promoter and the hygromycin phosphotransferase in some vermicelli products. Accordingly, we for the first time have detected the two transgenic Bt rice lines contaminating rice vermicelli samples. Furthermore, we developed a duplex real-time polymerase chain reaction (PCR) method for the simultaneous detection of both Bt rice lines. Publication Types: Research Support, Non-U.S. Gov't PMID: 17608495 [PubMed - indexed for MEDLINE] 424: J Agric Food Chem. 2007 Jul 25;55(15):6160-8. Epub 2007 Jul 3. Chemical composition of glyphosate-tolerant soybean 40-3-2 grown in Europe remains equivalent with that of conventional soybean (Glycine max L.). Harrigan GG, Ridley WP, Riordan SG, Nemeth MA, Sorbet R, Trujillo WA, Breeze ML, Schneider RW. Product Safety Center and Regulatory Affairs, Monsanto Company, 800 North Lindbergh Boulevard, St. Louis, Missouri 63167, USA. george.g.harrigan@monsanto.com The composition of glyphosate-tolerant (Roundup Ready) soybean 40-3-2 was compared with that of conventional soybean grown in Romania in 2005 as part of a comparative safety assessment program. Samples were collected from replicated field trials, and compositional analyses were performed to measure proximates (moisture, fat, ash, protein, and carbohydrates by calculation), fiber, amino acids, fatty acids, isoflavones, raffinose, stachyose, phytic acid, trypsin inhibitor, and lectin in grain as well as proximates and fiber in forage. The mean values for all biochemical components assessed for Roundup Ready soybean 40-30-2 were similar to those of the conventional control and were within the published range observed for commercial soybean. The compositional profile of Roundup Ready soybean 40-3-2 was also compared to that of conventional soybean varieties grown in Romania by calculating a 99% tolerance interval to describe compositional variability in the population of traditional soybean varieties already on the marketplace. These comparisons, together with the history of the safe use of soybean as a common component of animal feed and human food, lead to the conclusion that Roundup Ready soybean 40-3-2 is compositionally equivalent to and as safe and nutritious as conventional soybean varieties grown commercially. Publication Types: Comparative Study PMID: 17608426 [PubMed - indexed for MEDLINE] 425: Public Health Nutr. 2008 Jan;11(1):8-16. Epub 2007 Jul 3. Predictors of Australian consumers' intentions to consume conventional and novel sources of long-chain omega-3 fatty acids. Cox DN, Evans G, Lease HJ. CSIRO Food Futures National Research Flagship and CSIRO Human Nutrition, Adelaide BC, South Australia 5000, Australia. david.cox@csiro.au OBJECTIVES: To elicit predictors of variation in likelihood to purchase foods rich in long-chain omega-3 fatty acids. DESIGN, SETTING AND SUBJECTS: Responses from a community sample (n = 220) were elicited using a computer-administered questionnaire based on an adaptation of Protection Motivation Theory including measures of perceived risk and vulnerability to coronary heart disease (CHD). Other measures included health status, body mass index (BMI), perceived risk/benefits of novel technologies and sociodemographics. Descriptions of model products were presented, including farmed fish fed fishmeal (FFFF); farmed fish fed genetically modified (GM) oilseed (FFFGM); bread, milk and supplements containing fish oil (SFO) or GM oilseed. It was hypothesised that perceived vulnerability to CHD would enhance acceptance of GM products (H1). Furthermore, information describing the benefits of LCO3FA, limitations to fish supply and potential alternatives was given to a treatment group (50%) and hypothesised to have a positive effect on the acceptance of GM products (H2). RESULTS: No evidence was found to support H1 or H2. FFFF was most likely to be purchased (P < 0.01), followed by SFO and FFFGM. Multivariate regression analysis identified significant (P < 0.05) predictors (standardised beta) for likelihood to purchase FFFF: self-efficacy 0.56; behaviour (product) efficacy 0.19; belief that fishmeal is unnatural -0.14 (R2 = 0.44) and for FFFGM: self-efficacy 0.65; perceived severity of CHD 0.15; BMI -0.13; significant other has/had arthritis 0.11; belief that GM oilseed is unnatural 0.11 (R2 = 0.49). CONCLUSIONS: Self-efficacy (confidence to consume) was the most important predictor of likelihood to purchase all products. Publication Types: Research Support, Non-U.S. Gov't PMID: 17605836 [PubMed - indexed for MEDLINE] 426: Transgenic Res. 2008 Jun;17(3):367-77. Epub 2007 Jun 29. Assessment of the diversity and dynamics of Plum pox virus and aphid populations in transgenic European plums under Mediterranean conditions. Capote N, Pérez-Panadés J, Monzó C, Carbonell E, Urbaneja A, Scorza R, Ravelonandro M, Cambra M. Instituto Valenciano de Investigaciones Agrarias (IVIA), Carretera Moncada-Náquera km 5, 46113 Moncada, Valencia, Spain. The molecular variability of Plum pox virus (PPV) populations was compared in transgenic European plums (Prunus domestica L.) carrying the coat protein (CP) gene of PPV and non-transgenic plums in an experimental orchard in Valencia, Spain. A major objective of this study was to detect recombination between PPV CP transgene transcripts and infecting PPV RNA. Additionally, we assessed the number and species of PPV aphid vectors that visited transgenic and non-transgenic plum trees. Test trees consisted of five different P. domestica transgenic lines, i.e. the PPV-resistant C5 'HoneySweet' line and the PPV-susceptible C4, C6, PT6 and PT23 lines, and non-transgenic P. domestica and P. salicina Lind trees. No significant difference in the genetic diversity of PPV populations infecting transgenic and conventional plums was detected, in particular no recombinant between transgene transcripts and incoming viral RNA was found at detectable levels. Also, no significant difference was detected in aphid populations, including viruliferous individuals, that visited transgenic and conventional plums. Our data indicate that PPV-CP transgenic European plums exposed to natural PPV infection over an 8 year period caused limited, if any, risk beyond the cultivation of conventional plums under Mediterranean conditions in terms of the emergence of recombinant PPV and diversity of PPV and aphid populations. Publication Types: Comparative Study Research Support, Non-U.S. Gov't PMID: 17605085 [PubMed - indexed for MEDLINE] 427: Nature. 2007 Jun 28;447(7148):1042. Uganda hosts banana trial. Dauwers A. Publication Types: News PMID: 17597729 [PubMed - indexed for MEDLINE] 428: Physiol Behav. 2007 Nov 23;92(4):691-701. Epub 2007 May 21. Lentivirus-mediated downregulation of hypothalamic insulin receptor expression. Grillo CA, Tamashiro KL, Piroli GG, Melhorn S, Gass JT, Newsom RJ, Reznikov LR, Smith A, Wilson SP, Sakai RR, Reagan LP. Department of Pharmacology, Physiology and Neuroscience, University of South Carolina, Columbia, SC, United States. Regulation of feeding behavior and energy balance are among the central effects of insulin. For example, intracerebroventricular administration of insulin decreases food intake and body weight, whereas antisense oligodeoxynucleotide downregulation of insulin receptors (IRs) produces hyperphagia. To further examine the role of IRs in the central actions of insulin, we designed an IR antisense lentiviral vector (LV-IRAS) and injected this vector into the third ventricle to selectively decrease IR expression in the rat hypothalamus. Three weeks after LV-IRAS administration, the expression of IRs in the hypothalamus was significantly decreased, whereas no changes were observed in hippocampal IR levels. LV-IRAS administration decreased insulin-stimulated phosphorylation of hypothalamic IRs and translocation of the insulin-sensitive glucose transporter GLUT4 in the hypothalamus; no changes in IR signaling were observed in the hippocampus of LV-IRAS-treated rats. Lentivirus-mediated downregulation of IR expression and signaling produced significant increases in body weight, as well as increases in fat mass that were selective for the subcutaneous compartment. Conversely, lean muscle mass and water mass were not affected in LV-IRAS-treated rats compared to rats treated with control virus. Changes in peripheral adiposity were associated with increases in basal hypothalamic leptin signaling in the absence of changes in leptin receptor expression in LV-IRAS rats. Collectively, these data illustrate the important functional relationships between hypothalamic insulin and leptin signaling in the regulation of body composition and provide insight into the mechanisms through which decreases in IR expression and signaling dysregulates leptin activity, thereby promoting increases in peripheral adiposity. Publication Types: Evaluation Studies Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't PMID: 17585961 [PubMed - indexed for MEDLINE] 429: Indian J Exp Biol. 2007 Jun;45(6):554-62. Effect of Bt-cotton on chrysopids, ladybird beetles and their prey: aphids and whiteflies. Mellet MA, Schoeman AS. Department of Zoology & Entomology, University of Pretoria, Pretoria, 0002, South Africa. magdel.mellet@bayercropscience.com The effect of Bt-cotton, i.e. genetically modified cotton that contain genes expressing delta-endotoxin, on aphid, whitefly, chrysopid and coccinellid populations was determined with a two-year field study at a cotton farm near Marble Hall, South Africa. Although Bt-cotton is lepidopteran specific, non-lepidopteran arthropod populations may be indirectly influenced by the endotoxin. Abundance of aphid, whitefly, chrysopid and coccinellid populations and predator-prey interactions were used as measures to determine possible effects on the populations under investigation. The cultivation of Bt-cotton had no effect on aphid, whitefly, chrysopid or coccinellid abundance. Positive density dependent interactions occurred between aphids and coccinellids which were not influenced by Bt-cotton. A significant relationship between whitefly and coccinellid abundance, i.e. predator-prey reaction, occurred in the control and sprayed non-Bt cotton fields but was absent from the Bt-cotton fields. PMID: 17585692 [PubMed - indexed for MEDLINE] 430: Theriogenology. 2007 Sep 1;68 Suppl 1:S3-8. Epub 2007 Jun 15. Regulation of animal biotechnology: research needs. Rexroad CE Jr, Green RD, Wall RJ. U.S. Department of Agriculture, Agricultural Research Service, 5601 Sunnyside Avenue, Rm. 4-2150, Beltsville, MD 20705-5134, USA. caird.rexroad@ars.usda.gov Livestock that result from biotechnology have been a part of agricultural science for over 30 years but have not entered the market place as food or fiber. Two biotechnologies are at the forefront as challenges to the world's systems for regulating the market place: animal clones and transgenic animals. Both technologies have come before the Food and Drug Administration in the United States and it appears that action is imminent for clones. The FDA has asserted principles for evaluation of clones and asserts that "... remaining hazard(s) from cloning are likely to be subtle in nature." The science-based principles recognize that in some areas related to developmental biology and gene expression in clones, additional scientific information would be useful. The role of science then is to use the genomic tools that we have available to answer questions about epigenetic regulation of development and reprogramming of genes to the state found in germ cells. Transgenics pose additional challenges to regulators. If the transgenics are produced using cloning from modified cells then the additional scientific information needed will be related to the effects of insertion and expression of the transgenes. Other approaches such as retrovirally vectored transgenesis will elicit additional questions. These questions will be challenging because the science will have to be related to the expression and function of each gene or class of genes. For the promises of animal biotechnology to be fulfilled, scientists will have to resolve many questions for regulators and the public but tools to answer those questions are rapidly becoming available. Publication Types: Review PMID: 17574657 [PubMed - indexed for MEDLINE] 431: Nutr Cancer. 2007;58(1):66-74. Transgenic alfalfa that accumulates piceid (trans-resveratrol-3-O-beta-D-glucopyranoside) requires the presence of beta-glucosidase to inhibit the formation of aberrant crypt foci in the colon of CF-1 mice. Kineman BD, Au A, Paiva NL, Kaiser MS, Brummer EC, Birt DF. Department of Food Science and Human Nutrition, Iowa State University, Ames, IA 50011, USA. Plants have been genetically enhanced to produce a number of products for agricultural, industrial and pharmaceutical purposes. This technology could potentially be applied to providing chemoprevention strategies to the general population. Resveratrol (3,5,4'-trihydroxystilbene) is a compound that has been shown to have protective activity against a number of cancers and could be an ideal candidate for such an application. Alfalfa that was genetically modified to express resveratrol-synthase was used as a model in applying biotechnological approaches to cancer prevention. The transgenic alfalfa, which accumulates resveratrol as a glucoside (piceid = trans-resveratrol-3-O-Beta-D-glucopyranoside) (152 +/- 17.5 microg piceid/g dry weight), was incorporated into a standard mouse diet at 20% of the diet by weight and fed for 5 wk to 6-wk-old, female CF-1 mice (N = 17-30) that were injected with a single dose of azoxymethane (5 mg/kg body weight). While the addition of resveratrol-aglycone (20 mg/kg diet) to the basal diet reduced the number of aberrant crypt foci/mouse, the transgenic alfalfa did not inhibit the number, size, or multiplicity of aberrant crypt foci in the colon of the CF-1 mice relative to control alfalfa which does not accumulate resveratrol-glucoside. However, diets containing transgenic alfalfa with an exogenous Beta-glucosidase (860 U/kg diet) did significantly inhibit the number of aberrant crypt foci in the distal 2 cm of the colon of the mice relative to mice fed diets containing the transgenic alfalfa without the enzyme (P < 0.05; Fisher's Combination of p-values). The Beta-glucosidase alone appeared to have no effect on the inhibition of aberrant crypt foci. These results suggest that piceid in transgenic piceid-accumulating alfalfa was not bioavailable. Publication Types: Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. PMID: 17571969 [PubMed - indexed for MEDLINE] 432: Transgenic Res. 2008 Jun;17(3):345-54. Epub 2007 Jun 15. Interactions of Bacillus thuringiensis Cry1Ac toxin in genetically engineered cotton with predatory heteropterans. Torres JB, Ruberson JR. Departamento de Agronomia-Entomologia, Universidade Federal Rural de Pernambuco, Av. Dom Manoel de Medeiros, s/n, Dois Irmãos, 52171-900 Recife, Pernambuco, Brasil. jtorres@depa.ufrpe.br A number of cotton varieties have been genetically transformed with genes from Bacillus thuringiensis (Bt) to continuously produce Bt endotoxins, offering whole plant and season-long protection against many lepidopteran larvae. Constant whole-plant toxin expression creates a significant opportunity for non-target herbivores to acquire and bio-accumulate the toxin for higher trophic levels. In the present study we investigated movement of Cry1Ac toxin from the transgenic cotton plant through specific predator-prey pairings, using omnivorous predators with common cotton pests as prey: (1) the beet armyworm, Spodoptera exigua (Lepidoptera: Noctuidae), with the predator Podisus maculiventris (Heteroptera: Pentatomidae); (2) the two-spotted spider mite, Tetranychus urticae (Acarina: Tetranychidae), with the predatory big-eyed bug Geocoris punctipes (Heteroptera: Geocoridae) and (3) with the predatory damsel bug Nabis roseipennis (Heteropera: Nabidae); and (4) the thrips Frankliniella occidentalis (Thysanoptera: Thripidae) with the predatory pirate bug Orius insidiosus (Heteroptera: Anthocoridae). We quantified Cry1Ac toxin in the cotton plants, and in the pests and predators, and the effects of continuous feeding on S. exigua larvae fed either Bt or non-Bt cotton on life history traits of P. maculiventris. All three herbivores were able to convey Cry1Ac toxin to their respective predators. Among the herbivores, T. urticae exhibited 16.8 times more toxin in their bodies than that expressed in Bt-cotton plant, followed by S. exigua (1.05 times), and F. occidentalis immatures and adults (0.63 and 0.73 times, respectively). Of the toxin in the respective herbivorous prey, 4, 40, 17 and 14% of that amount was measured in the predators G. punctipes, P. maculiventris, O. insidiosus, and N. roseipennis, respectively. The predator P. maculiventris exhibited similar life history characteristics (developmental time, survival, longevity, and fecundity) regardless of the prey's food source. Thus, Cry1Ac toxin is conveyed through non-target herbivores to natural enemies at different levels depending on the herbivore species, but continuous lifetime contact with the toxin by the predator P. maculiventris through its prey had no effect on the predator's life history. The results found here, supplemented with others already published, suggest that feeding on Cry1Ac contaminated non-target herbivores does not harm predatory heteropterans and, therefore, cultivation of Bt cotton may provide an opportunity for conservation of these predators in cotton ecosystems by reducing insecticide use. Publication Types: Research Support, Non-U.S. Gov't PMID: 17570072 [PubMed - indexed for MEDLINE] 433: J Biotechnol. 2007 Jun 30;130(3):236-46. Epub 2007 Apr 24. Regulation of alcohol oxidase of a recombinant Pichia pastoris Mut+ strain in transient continuous cultures. Jungo C, Marison I, von Stockar U. Ecole Polytechnique Fédérale de Lausanne, Laboratoire de Génie Chimique et Biologique, Station 6, CH-1015 Lausanne, Switzerland. In the methylotrophic yeast Pichia pastoris, alcohol oxidase (AOX) is a key enzyme involved in the dissimilation of methanol. Heterologous proteins are usually expressed under the control of the AOX1 promoter, which drives the expression of alcohol oxidase 1 in the wild-type strain. This study investigates the regulation of the alcohol oxidase enzyme of a recombinant P. pastoris Mut+ strain in cultures on glycerol and methanol as sole carbon sources and in mixed substrate cultures on both substrates. The aim was to have a better insight in the transition from growth on glycerol to growth on methanol, which is a key step in standard high cell density P. pastoris cultures for the production of foreign proteins. Nutrient shifts in chemostat cultures showed that after growth on glycerol use of mixed feeds of glycerol and methanol allowed faster induction of alcohol oxidase and faster adaptation of cellular metabolism than with a feed containing methanol as sole carbon source. The results of this study showed also how critical it is to avoid transient methanol accumulation during P. pastoris cultures operated at low residual methanol concentrations. Indeed, pulse experiments during chemostat cultures showed that sudden increase in methanol concentrations in cultures performed under methanol-limited or dual methanol and glycerol-limited growth conditions leads to wash-out of the culture because of too high consumption rate of methanol, which leads to excretion of toxic intermediates. High rate of methanol consumption was due to high specific AOX activities observed at low residual methanol concentrations. Publication Types: Research Support, Non-U.S. Gov't PMID: 17566583 [PubMed - indexed for MEDLINE] 434: Br J Soc Psychol. 2007 Jun;46(Pt 2):437-57. Predicting behaviour towards genetically modified food using implicit and explicit attitudes. Spence A, Townsend E. RASPH, School of Psychology, University of Nottingham, UK. spenceAl@cardiff.ac.uk The predictive validity of implicit and explicit attitudes is a central question in social psychological research with important theoretical and empirical ramifications. Three main patterns of combining implicit and explicit attitudes to predict behaviour have been postulated. They are, double dissociation (in which implicit and explicit attitudes predict spontaneous and deliberate behaviour respectively), additive (in which implicit and explicit attitudes both predict variance in behaviour) and interactive (in which implicit and explicit attitudes combine to predict behaviour). These models were tested in this study using a structural equation modelling approach utilising three different measures of behaviour (of varying spontaneity) towards genetically modified (GM) food. The additive pattern, in which implicit and explicit attitudes both predict variance in behaviour, was found to best fit the data. In addition, all behaviour measures indicated that the majority of participants were willing to try GM food in some situations. Publication Types: Research Support, Non-U.S. Gov't PMID: 17565791 [PubMed - indexed for MEDLINE] 435: Infect Genet Evol. 2008 Jul;8(4):520-5. Epub 2007 May 6. From population structure to genetically-engineered vectors: new ways to control vector-borne diseases? Sparagano OA, De Luna CJ. School of Agriculture, Food, and Rural Development, Agriculture Building, Newcastle University, Newcastle upon Tyne, UK. Olivier.Sparagano@ncl.ac.uk Epidemiological studies on vectors and the pathogens they can carry (such as Borrelia burgdorferi) are showing some correlations between infection rates and biodiversity highlighting the "dilution" effects on potential vectors. Meanwhile other studies comparing sympatric small rodent species demonstrated that rodent species transmitting more pathogens are parasitized by more ectoparasite species. Studies on population structure and size have also proven a difference on the intensity of the parasitic infection. Furthermore, preliminary results in genetic improvement in mosquitoes (genetic markers, sexing, and genetic sterilization) will also increase performance as it has already been shown in field applications in developing countries. Recent results have greatly improved the fitness of genetically-modified insects compared to wild type populations with new approaches such as the post-integration elimination of transposon sequences, stabilising any insertion in genetically-modified insects. Encouraging results using the Sterile Insect Technique highlighted some metabolism manipulation to avoid the viability of offspring from released parent insect in the wild. Recent studies on vector symbionts would also bring a new angle in vector control capabilities, while complete DNA sequencing of some arthropods could point out ways to block the deadly impact on animal and human populations. These new potential approaches will improve the levels of control or even in some cases would eradicate vector species and consequently the vector-borne diseases they can transmit. In this paper we review some of the population biology theories, biological control methods, and the genetic techniques that have been published in the last years that are recommended to control for vector-borne diseases. Publication Types: Review PMID: 17560836 [PubMed - indexed for MEDLINE] 436: J Biotechnol. 2007 Oct 31;132(2):180-6. Epub 2007 Apr 29. Process Analytical Technology (PAT): batch-to-batch reproducibility of fermentation processes by robust process operational design and control. Gnoth S, Jenzsch M, Simutis R, Lübbert A. Center for Bioprocess Engineering, Martin-Luther-University Halle-Wittenberg, Germany. The Process Analytical Technology (PAT) initiative of the FDA is a reaction on the increasing discrepancy between current possibilities in process supervision and control of pharmaceutical production processes and its current application in industrial manufacturing processes. With rigid approval practices based on standard operational procedures, adaptations of production reactors towards the state of the art were more or less inhibited for long years. Now PAT paves the way for continuous process and product improvements through improved process supervision based on knowledge-based data analysis, "Quality-by-Design"-concepts, and, finally, through feedback control. Examples of up-to-date implementations of this concept are presented. They are taken from one key group of processes in recombinant pharmaceutical protein manufacturing, the cultivations of genetically modified Escherichia coli bacteria. Publication Types: Research Support, Non-U.S. Gov't PMID: 17559961 [PubMed - indexed for MEDLINE] 437: Plant J. 2007 Jul;51(2):165-72. Epub 2007 Jun 8. Viral infection enables phloem loading of GFP and long-distance trafficking of the protein. Peleg G, Malter D, Wolf S. Institute of Plant Sciences and Genetics in Agriculture and Otto Warburg Minerva Center for Agricultural Biotechnology, The Hebrew University of Jerusalem, Faculty of Agricultural, Food and Environmental Quality Sciences, Rehovot, Israel. It is generally accepted that viral systemic infection follows the source-to-sink symplastic pathway of sugar translocation. In plants that are classified as apoplastic loaders, the boundary between the companion cell-sieve element (CC-SE) complex and neighboring cells is symplastically restricted, and the potential passage of macromolecules between the two domains has yet to be explored. Transgenic tobacco plants expressing green fluorescence protein (GFP) and cucumber mosaic virus (CMV)-encoded proteins fused to GFP under the control of the fructose-1,6-bisphosphatase (FBPase) promoter were produced in order to localize the encoded proteins in mesophyll and bundle sheath cells and to explore the influence of viral infection on the functioning of plasmodesmata interconnecting the two domains. GFP produced outside the vascular tissue could overcome the symplastic barrier between the CC-SE complex and the surrounding cells to enter the vasculature in CMV-infected plants. Grafting of control (non-transgenic) tobacco scions to CMV-infected FBPase-GFP-expressing root stocks confirmed that GFP could move long distances in the phloem. No movement of the gfp mRNA was noticeable in this set of experiments. The ability of GFP to enter the vasculature and move long distances was also evident upon infection of the grafting plants with other viruses. These results provide experimental evidence for alteration of the functioning of plasmodesmata interconnecting the CC-SE complex and neighboring cells by viral infection to enable non-selective trafficking of macromolecules from the mesophyll into the sieve tube. Publication Types: Research Support, Non-U.S. Gov't PMID: 17559510 [PubMed - indexed for MEDLINE] 438: Crit Rev Food Sci Nutr. 2007;47(5):441-98. Implementation of physicochemical and sensory analysis in conjunction with multivariate analysis towards assessing olive oil authentication/adulteration. Arvanitoyannis IS, Vlachos A. University of Thessaly, School of Agricultural Sciences, Department of Agriculture Animal Production and Aquatic Environment, Volos, Hellas, Greece. parmenion@uth.gr The authenticity of products labeled as olive oils, and in particular as virgin olive oils, stands for a very important issue both in terms of its health and commercial aspects. In view of the continuously increasing interest in virgin olive oil therapeutic properties, the traditional methods of characterization and physical and sensory analysis were further enriched with more advanced and sophisticated methods such as HPLC-MS, HPLC-GC/C/IRMS, RPLC-GC, DEPT, and CSIA among others. The results of both traditional and "novel" methods were treated both by means of classical multivariate analysis (cluster, principal component, correspondence, canonical, and discriminant) and artificial intelligence methods showing that nowadays the adulteration of virgin olive oil with seed oil is detectable at very low percentages, sometimes even at less than 1%. Furthermore, the detection of geographical origin of olive oil is equally feasible and much more accurate in countries like Italy and Spain where databases of physical/chemical properties exist. However, this geographical origin classification can also be accomplished in the absence of such databases provided that an adequate number of oil samples are used and the parameters studied have "discriminating power." Publication Types: Review PMID: 17558656 [PubMed - indexed for MEDLINE] 439: Nat Biotechnol. 2007 Jun;25(6):624-6. Comment on: Nat Biotechnol. 2006 May;24(5):498; author reply 499. Trends in GM crop, food and feed safety literature. Vain P. Publication Types: Comment Letter PMID: 17557092 [PubMed - indexed for MEDLINE] 440: Wei Sheng Yan Jiu. 2007 Mar;36(2):245-8. [Supervision of genetically modified foods by the international community] [Article in Chinese] Wang R, Yang X. Institute for Nutrition and Food Safety, Chinese Center for Disease Control and Prevention, Beijing 100050, China. The genetically modified foods (GMF) are latent with great commercial potentiality and related with public health. The international organizations and the governments have been attached important to it and constituted correlative statutes. The article is intended to introduce the development of GMF, review some correlative statutes about GMF supervision by the international organizations and the governments. It is significant to constitute and consummate the law system of GMF in our country. Publication Types: English Abstract PMID: 17555112 [PubMed - in process] 441: Plant Cell Rep. 2007 Oct;26(10):1821-31. Epub 2007 Jun 7. Event-specific qualitative and quantitative PCR detection of roundup ready event GT73 based on the 3'-integration junction. Yang R, Xu W, Luo Y, Guo F, Lu Y, Huang K. College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, China. With the development of genetically modified organisms, labeling regulations have been introduced, which require appropriate detection methods. Event-specific qualitative and quantitative polymerase chain reaction (PCR) detection methods have become the internationally agreed state-of-art. This paper describes an event-specific PCR method for qualitative and quantitative of Roundup Ready canola event GT73. The 3'-integration junction was characterized by two methods: inverse-PCR and thermal asymmetric interlaced-PCR. In the conventional qualitative PCR assay, the event-specific primers designed were confirmed to be specific and the limit of detection (LOD) was 0.05% (approximates to ten haploid genome copies). In the quantitative TaqMan real-time PCR assay, the LOD and the limit of quantification were five and ten haploid genome copies, respectively. In addition, for further quantitative detection, a reference molecule which contained the canola endogenous gene and event-specific sequence was constructed and standard curves were set up. The goodness of the linearity and high efficiency of the PCR reaction indicated the usability of the plasmid and the established PCR system. Moreover, mixed samples with different GT73 content (6, 3, 1 and 0.5%) were quantified using the established real-time PCR system to evaluate the trueness and precision of the system. The trueness expressed as bias varied from 2.00 to 18.00%. The precision expressed as variation coefficient were different from 6.40 to 32.95%. From above results, we believed that the established event-specific qualitative and quantitative PCR systems for GT73 in this study were acceptable and suitable for genetic modified canola detection. Publication Types: Research Support, Non-U.S. Gov't PMID: 17554542 [PubMed - indexed for MEDLINE] 442: Med Hypotheses. 2007;69(6):1257-60. Epub 2007 Jun 5. Are xenogeneic anti-tissue transglutaminase antibodies the holy grail for celiac patients? Ivanovski PI, Ivanovski IP, Sedlarevic R. University Children's Hospital, 10 Tirshova Str., Belgrade, Serbia. ivanovsk@eunet.yu Celiac disease is an immune mediated disorder, the only one with a well-established origin, resulting from a permanent gluten intolerance. Although a gluten-free diet is currently the "safe" and appropriate therapy for celiac disease, this is not always an easy and simple option as "harmful" gluten may contaminate food during the processing and preparation phases. There are also further social pressures, which might be more pressing for young celiac patients, in following a strict gluten-free diet. Therefore, a new therapeutic approaches are sought which would permit celiacs to "peacefully" coexist with gluten. Presently, the most promising looks search for genetically modified wheat lacking toxic gluten peptides and the use of oral endopeptidases in attempt to curb gluten toxicity. Recently discovered role of anti-tissue transglutaminase antibodies in celiac pathogenesis has brought a prospect for a new hypothetical therapeutic approach, an oral immunization of celiacs with xenogeneic anti-tissue transglutaminase antibodies. PMID: 17553630 [PubMed - indexed for MEDLINE] 443: Food Nutr Bull. 2006 Sep;27(3):265-6. Quality protein maize. Scrimshaw NS. PMID: 17542118 [PubMed - indexed for MEDLINE] 444: Rev Biol Trop. 2007 Jun;55(2):347-64. [Genetically modified crops: promises and good intentions are not enough (refutation to Espinoza et aL 2004, Rev. Biol. Trop. 52 (3): 727-732)] [Article in Spanish] García JE. Centro de Educación Ambiental de la Universidad Estatal a Distancia y Escuela de Biologia de la Universidad de Costa Rica, San José, Costa Rica. jaimeenrique56@yahoo.com The arguments presented by Espinoza et al. in their paper "Relationship of genetically modified crops with the environment and health of the Costa Rican human population" published in this journal (Rev. Biol. Trop. 52: 727-732, 2004) are questioned and refuted. The arguments are confronted with evidence offered by scientists and national and international independent organizations around the world (e.g. World Health Organization, Consumers International, Physicians and Scientists for Responsible Application of Science and Technology, International Union for Conservation of Nature and Natural Resources, the Council of the University of Costa Rica, and the Independent Science Panel) showing the current uncertainty and limitations of science in this area, as well as those of proposed and applied biosafety approaches. Environment, biodiversity and food security are so important and basic matters, that there is need of serious testing, particularly when promises seem to be based on environmentally dangerous ideas promoted half a century ago by the so called "green revolution". Debate should continue, based on a holistic analysis of facts and with ethical reasoning, avoiding emotional positions that can confuse virtual reality with reality. Publication Types: Comment English Abstract PMID: 19069750 [PubMed - in process] 445: Pest Manag Sci. 2007 Jul;63(7):658-76. Mites for the control of pests in protected cultivation. Gerson U, Weintraub PG. Department of Entomology, Faculty of Agricultural, Food and Environmental Quality Sciences, Rehovot 76100, Israel. The production of crops under protected conditions is increasing worldwide. Owing to growing consumer demands for healthy and green produce, and intensifying pesticide resistance, non-chemical solutions--foremost among which is biological control--are being sought. The authors review recent advances related to the application of predatory mites for the control of greenhouse pests, and discuss interactions among acarine biocontrol agents (ABAs) and the effects of crop plants and new technologies on ABAs, such as artificial lighting, elevated carbon dioxide levels and genetically modified organisms. This is followed by a discussion of the problems associated with the search for and use of new ABAs, including management, the benefits of modelling and avenues of future research. Copyright (c) 2007 Society of Chemical Industry. Publication Types: Review PMID: 17533640 [PubMed - indexed for MEDLINE] 446: Langmuir. 2007 Jun 19;23(13):7189-95. Epub 2007 May 26. Adsorption to metal oxides of the Pseudomonas aeruginosa siderophore pyoverdine and implications for bacterial biofilm formation on metals. Upritchard HG, Yang J, Bremer PJ, Lamont IL, McQuillan AJ. Departments of Biochemistry, Chemistry, and Food Science, University of Otago, Dunedin, New Zealand. The initiation of biofilm formation is poorly understood, and in particular, the contribution of chemical bond formation between bacterial cells and metal surfaces has received little attention. We have previously used in situ infrared spectroscopy to show, during the initial stages of Pseudomonas aeruginosa biofilm formation, the formation of coordinate covalent bonds between titanium dioxide particle films and pyoverdine, a mixed catecholate and hydroxamate siderophore. Here we show using infrared spectroscopy that pyoverdine can also form covalent bonds with particle films of Fe2O3, CrOOH, and AlOOH. Adsorption to the metal oxides through the catechol-like 2,3-diamino-6,7-dihydroxyquinoline part of pyoverdine was most evident in the infrared spectrum of the adsorbed pyoverdine molecule. Weaker infrared absorption bands that are consistent with the hydroxamic acids of pyoverdine binding covalently to TiO2, Fe2O3, and AlOOH surfaces were also observed. The adsorption of pyoverdine to TiO2 and Fe2O3 surfaces showed a pH dependence that is indicative of the dominance of the catechol-like ligand of pyoverdine. Infrared absorption bands were also evident for pyoverdine associated with the cells of P. aeruginosa on TiO2 and Fe2O3 surfaces and were notably absent for genetically modified cells unable to synthesize or bind pyoverdine at the cell surface. These studies confirm the generality of pyoverdine-metal bond formation and suggest a wider involvement of siderophores in bacterial biofilm initiation on metals. Publication Types: Research Support, Non-U.S. Gov't PMID: 17530790 [PubMed - indexed for MEDLINE] 447: Biotechnol J. 2007 Jul;2(7):826-32. Safety assessment of genetically modified organisms of plant origin in the Russian Federation. Tyshko NV, Aksyuk IN, Tutelyan VA. Institute of Nutrition, Russian Academy of Medical Sciences, Moscow, Russian Federation. The beginning of the 21st century is characterized by growing interest in the problems of biosafety, which are determined, on the one hand, by the wide use of novel biotechnologies and the necessity to develop the adequate precautionary measures, and, on the other hand, by the objective threat of bioterrorism. Therefore, improvement of the estimation system for genetically modified (GM) sources of food and strengthening the control of their circulation are the urgent problems of modern biology and medicine. Russia is one of the countries where the estimation system of food products obtained from the GM sources is rather efficient. The key features of this system are the complex toxicological and epidemiological examinations. One of the main parts of GM food safety assessment is based upon detection of their potentially toxic properties, which could provoke unintended effects of the genetic modification. Publication Types: Review PMID: 17526054 [PubMed - indexed for MEDLINE] 448: Bull Entomol Res. 2007 Jun;97(3):265-80. Statistical models to evaluate invertebrate-plant trophic interactions in arable systems. Bohan DA, Hawes C, Haughton AJ, Denholm I, Champion GT, Perry JN, Clark SJ. Rothamsted Research, Harpenden, Herts, AL5 2JQ, UK. David.Bohan@bbrsc.ac.uk Over the past 40 years there have been marked shifts in arable farmland management that are widely believed to have had a considerable impact on flowering plants and invertebrates and the small mammals and birds that rely upon them. It is not yet possible to predict the dynamics of plants and invertebrates either with past or future changes in farmland management. This study investigates whether a basic invertebrate classification, formed of broad trophic groups, can be used to describe interactions between invertebrates and their resource plants and evaluate management impacts for genetically modified, herbicide-tolerant (GMHT) and conventional herbicide management in both spring- and winter-sown oilseed rape. It is argued that the analyses validate trophic-based approaches for describing the dynamics of invertebrates in farmland and that linear models might be used to describe the changes in invertebrate trophic group abundance in farmland when driven by primary producer abundance or biomass and interactions between invertebrates themselves. The analyses indicate that invertebrate dynamics under GMHT management are not unique, but similar to conventional management occurring over different resource ranges, and that dynamics differed considerably between spring- and winter-sown oilseed rape. Thus, herbicide management was of much lower impact on trophic relationships than sowing date. Results indicate that invertebrate dynamics in oilseed rape are regulated by a combination of top-down and bottom-up trophic processes. Publication Types: Research Support, Non-U.S. Gov't Validation Studies PMID: 17524158 [PubMed - indexed for MEDLINE] 449: Adv Biochem Eng Biotechnol. 2007;107:133-51. Prospects for biopolymer production in plants. van Beilen JB, Poirier Y. Département de Biologie Moléculaire Végétale, Université de Lausanne, Bâtiment Biophore, 1015, Lausanne, Switzerland. It is likely that during this century polymers based on renewable materials will gradually replace industrial polymers based on petrochemicals. This chapter gives an overview of the current status of research on plant biopolymers that are used as a material in non-food applications. We cover technical and scientific bottlenecks in the production of novel or improved materials, and the potential of using transgenic or alternative crops in overcoming these bottlenecks. Four classes of biopolymers will be discussed: starch, proteins, natural rubber, and poly-beta-hydroxyalkanoates. Renewable polymers produced by chemical polymerization of monomers derived from sugars, vegetable oil, or proteins, are not considered here. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 17522824 [PubMed - indexed for MEDLINE] 450: Adv Biochem Eng Biotechnol. 2007;107:57-68. Genetically modified organisms in the United States: implementation, concerns, and public perception. Oeschger MP, Silva CE. Department of Microbiology, Immunology and Parasitology, Louisiana State University Health Sciences Center, New Orleans, LA 70112, USA. moesch@lsuhsc.edu We examine the state of biotechnology with respect to genetically modified (GM) organisms in agriculture. Our focus is on the USA, where there has been significant progress and implementation but where, to date, the matter has drawn little attention. GM organisms are the result of lateral gene transfers, the transfer of genes from one species to another, or sometimes, from one kingdom to another. The introduction of foreign genes makes some people very uncomfortable, and a small group of activists have grave concerns about the technology. Attempts by activists to build concern in the general public have garnered little attention; however, the producers of GM organisms have responded to their concerns and established extensive testing programs to be applied to each candidate organism that is produced. In the meantime, GM varieties of corn, cotton, soybean and rapeseed have been put into agricultural production and are now extensively planted. These crops, and the other, newer GM crops, have produced no problems and have pioneered a silent agricultural revolution in the USA. Publication Types: Review PMID: 17522820 [PubMed - indexed for MEDLINE] 451: Adv Biochem Eng Biotechnol. 2007;107:1-11. The gap between science and perception: the case of plant biotechnology in Europe. Einsele A. Internutrition, Postfach, 8035, Zurich, Switzerland. arthur.einsele@internutrition.ch Although the global area of biotech crops continues to climb for the tenth consecutive year at a sustainable double-digit growth rate, the acceptance of biotech products from agriculture in Europe is still low. There is a gap between science and perception. It is a strong belief that the public turning against science and against GM food has been encouraged by the negative activities of NGO groups. Scientists have to overcome the purely risk-based discussion, and the benefits of plant biotechnology have to be made literally visible. GM food should be available, the benefits should be tangible and the consumer should have fun with such novel food. The gap could be reduced if genetically modified plants and the products thereof were regulated in the same way as classical products. Publication Types: Review PMID: 17522817 [PubMed - indexed for MEDLINE] 452: Clin Exp Allergy. 2007 Jun;37(6):918-28. Genetically glycosylated ovomucoid third domain can modulate Immunoglobulin E antibody production and cytokine response in BALB/c mice. Rupa P, Nakamura S, Mine Y. Department of Food Science, University of Guelph, Guelph, ON, Canada N1G 2W1. BACKGROUND: Food allergies are on the rise and it is estimated that in North America, 8% of the children and 4% of the adults have food allergies. Food allergies tend to occur more often in children than in adults due to their immature digestive and immune systems. Hen's egg is among the most common cause of food-induced allergic reactions in North America. OBJECTIVE: The present study was undertaken to investigate the role of N-glycans of the third domain of ovomucoid in IgE binding and modulation of allergen-specific immune response in BALB/c mice. METHODS: The cDNA encoding the third domain of ovomucoid was inserted into the yeast genome and expressed in Pichia pastoris X-33 cells, under the control of the glyceraldehyde-3-phosphate (GAP) dehydrogenase promoter for constitutive expression to obtain a post-translationally modified and functionally active ovomucoid third domain. Upon expression, the protein was secreted into the extracellular medium and was purified by size exclusion chromatography. The recombinant protein was produced at 10 mg/L of the culture supernatant. BALB/c mice were sensitized with the recombinant and native forms of glycosylated ovomucoid third domain antigen. The allergic response of the native and the recombinant glycosylated forms of ovomucoid third domain antigens were compared using antibody and cytokine measurements. RESULTS: ELISA tests indicated a significant decrease in specific IgE antibodies to the recombinant N-linked glycosylated form (P-Gly), when compared with the native glycosylated form (DIII+) using mice sera. Immunization with P-Gly induced the production of IFN-gamma [T-helper type 1 (Th1) response] and lowered the production of IL-4 (Th2 response), and a skewed balance towards the Th1 cytokine demonstrated that P-Gly has a modulating ability on Th1/Th2 balance to down-regulate Th2 response. Furthermore, N-linked glycan (N28) in the third domain of ovomucoid was shown to be associated with suppression of the allergic response. CONCLUSION: Therefore, we can conclude that P-Gly facilitates and contributes to the discovery of new molecular target for the development of a safe and specific therapeutic vaccine for the treatment of egg allergy, and oligosaccharides do seem to play a major role in the suppression of IgE-binding activity. Publication Types: Comparative Study Research Support, Non-U.S. Gov't PMID: 17517106 [PubMed - indexed for MEDLINE] 453: J Agric Food Chem. 2007 Jun 13;55(12):4728-34. Epub 2007 May 18. Development of a certified reference material for genetically modified potato with altered starch composition. Broothaerts W, Corbisier P, Emons H, Emteborg H, Linsinger TP, Trapmann S. European Commission, Joint Research Centre, Institute for Reference Materials and Measurements (IRMM), Retieseweg 111, 2440 Geel, Belgium. Wim.Broothaerts@ec.europa.eu The presence of genetically modified organisms (GMOs) in food and feed products is subject to regulation in the European Union (EU) and elsewhere. As part of the EU authorization procedure for GMOs intended for food and feed use, reference materials must be produced for the quality control of measurements to quantify the GMOs. Certified reference materials (CRMs) are available for a range of herbicide- and insect-resistant genetically modified crops such as corn, soybean, and cotton. Here the development of the first CRM for a GMO that differs from its non-GMO counterpart in a major compositional constituent, that is, starch, is described. It is shown that the modification of the starch composition of potato (Solanum tuberosum L.) tubers, together with other characteristics of the delivered materials, have important consequences for the certification strategy. Moreover, the processing and characterization of the EH92-527-1 potato material required both new and modified procedures, different from those used routinely for CRMs produced from genetically modified seeds. PMID: 17508757 [PubMed - indexed for MEDLINE] 454: J Agric Food Chem. 2007 Jun 13;55(12):4645-50. Epub 2007 May 16. Quantification of chlorophyll content and classification of nontransgenic and transgenic tomato leaves using visible/near-infrared diffuse reflectance spectroscopy. Xie L, Ying Y, Ying T. College of Biosystems Engineering and Food Science, Zhejiang University, 268 Kaixuan Street, 310029 Hangzhou, People's Republic of China. Visible/near-infrared (vis/NIR) spectroscopy combined with multivariate analysis was used to quantify chlorophyll content in tomato leaves and classify tomato leaves with different genes. In this study, transgenic tomato leaves with antisense LeETR1 (n = 106) and their parent nontransgenic ones (n = 102) were measured in vis/NIR diffuse reflectance mode. Quantification of chlorophyll content was achieved by partial least-squares regression with a cross-validation prediction error equal to 2.87. Partial least-squares discriminant analysis was performed to classify leaves. The results show that differences between transgenic and nontransgenic tomato leaves do exist, and excellent classification can be obtained after optimizing spectral pretreatment. The classification accuracy can reach to 100% using the derivative of spectral data in the full and partial wavenumber range. These results demonstrate that vis/NIR spectroscopy together with chemometrics techniques could be used to quantify chlorophyll content and differentiate tomato leaves with different genes, which offers the benefit of avoiding time-consuming, costly, and laborious chemical and sensory analysis. Publication Types: Research Support, Non-U.S. Gov't PMID: 17503831 [PubMed - indexed for MEDLINE] 455: Int Arch Allergy Immunol. 2007;144(1):29-38. Epub 2007 May 11. A proteomic study to identify soya allergens--the human response to transgenic versus non-transgenic soya samples. Batista R, Martins I, Jeno P, Ricardo CP, Oliveira MM. Instituto Nacional de Saúde Dr. Ricardo Jorge, Lisboa, Portugal. rita.batista@insa.min-saude.pt BACKGROUND: In spite of being among the main foods responsible for allergic reactions worldwide, soybean (Glycine max)-derived products continue to be increasingly widespread in a variety of food products due to their well-documented health benefits. Soybean also continues to be one of the elected target crops for genetic modification. The aim of this study was to characterize the soya proteome and, specifically, IgE-reactive proteins as well as to compare the IgE response in soya-allergic individuals to genetically modified Roundup Ready soya versus its non-transgenic control. METHODS: We performed two-dimensional gel electrophoresis of protein extracts from a 5% genetically modified Roundup Ready flour sample and its non-transgenic control followed by Western blotting with plasma from 5 soya-sensitive individuals. We used peptide tandem mass spectrometry to identify soya proteins (55 protein matches), specifically IgE-binding ones, and to evaluate differences between transgenic and non-transgenic samples. RESULTS: We identified 2 new potential soybean allergens--one is maturation associated and seems to be part of the late embryogenesis abundant proteins group and the other is a cysteine proteinase inhibitor. None of the individuals tested reacted differentially to the transgenic versus non-transgenic samples under study. CONCLUSION: Soybean endogenous allergen expression does not seem to be altered after genetic modification. Proteomics should be considered a powerful tool for functional characterization of plants and for food safety assessment. Copyright (c) 2007 S. Karger AG, Basel. Publication Types: Comparative Study Research Support, Non-U.S. Gov't PMID: 17496424 [PubMed - indexed for MEDLINE] 456: ALTEX. 2006;23 Suppl:284-7. Safety Assessment of Genetically Modified (GM) Foods. O'mahony P, Reilly A. Food Safety Authority of Ireland, Dublin, Ireland. Modern biotechnology enables a transfer of genes between species that would not occur naturally. Safety assessment of genetically modified (GM) foods is primarily based on the concept of substantial equivalence, a comparison with conventional counterparts and similar varieties as a starting point. Nutritional and toxicological studies should be based on the outcome of the comparison, and additional safety tests should be carried out as required. The limitations of animal studies, particularly in assessing the safety of whole GM foods may be addressed by the new technologies that brought us GM foods in the first place. While animal studies may still have a role in the testing of GM food components, the advent of technology such as "genomics" offers a real and possibly superior alternative. PMID: 17492196 [PubMed - in process] 457: Xenotransplantation. 2007 May;14(3):217-21. Some ethical issues regarding xenotransfusion. Roux FA, Saï P, Deschamps JY. Department of Cellular and Molecular Immuno-Endocrinology, INRA, Nantes School of Veterinary Medicine, Atlanpole, La Chantrerie, Nantes, France. BACKGROUND: The use of porcine red blood cells has recently been proposed as a possible solution to the shortage of blood for human transfusion. OBJECTIVES: The purpose of this paper is to compare some ethical issues regarding xenotransfusion (XTF) with those relating to xenotransplantation (XT) of organs, tissues and cells. MATERIALS AND METHODS: Various ethical concerns and viewpoints relating to XTF are discussed. RESULTS: The main ethical obstacles to XT do not apply to XTF. It is much more ethically acceptable to raise pigs for regular blood collection as it doesn't damage the health of the animal. Porcine endogenous retrovirus infection, the major concern associated with XT, does not apply to XTF, since red blood cells have no DNA and have a very short lifespan. Clinical trials will be possible in humans once XTF has been demonstrated to be effective and harmless in non-human primates. Transgenesis is acceptable for pig blood donors because only a limited number of genes are involved, and these animals will never enter into the livestock gene pool or the food chain. CONCLUSION: Because the need for blood is less pressing than that for organs, tissues or cells, the use of animal blood for human transfusion is not an absolute necessity. However, it represents a real opportunity. The ability to gain access to an unlimited quantity of blood is a reasonable justification for XTF. Because its technical and ethical hurdles are less stringent, XTF could be the first large-scale clinical application of XT. PMID: 17489861 [PubMed - indexed for MEDLINE] 458: J Agric Food Chem. 2007 Jun 13;55(12):4670-7. Epub 2007 May 10. Recombinant porcine lactoferrin expressed in the milk of transgenic mice enhances offspring growth performance. Wu SC, Chen HL, Yen CC, Kuo MF, Yang TS, Wang SR, Weng CN, Chen CM, Cheng WT. Department of Animal Science and Technology, National Taiwan University, Taipei 106, Taiwan. The European Commission has proposed a permanent ban on the use of antibiotics as an ingredient in animal feed to promote growth. Lactoferrin is a globular multifunctional protein that has been shown to play a role in iron absorption and to have antimicrobial and anti-inflammatory activities. Therefore, lactoferrin may serve as a nontherapeutic alternative to antibiotics in livestock husbandry. As a pilot study toward this goal, transgenic mice have been generated harboring a porcine lactoferrin (pLF) gene driven by the mammary gland-specific promoter of the bovine alpha-lactalbumin (alphaLA) gene. The alphaLA-pLF hybrid gene was confirmed to have been successfully integrated and transmitted stably through the germ-line in 9 (5 females and 4 males) of 14 transgenic founders. In the female progenies of six lines analyzed, the transgene copy numbers ranged from 1 to 20 with 1-4 integration sites. Significant levels of pLF protein in milk ranging from 40 to 106 microg/mL with physical characteristics similar to those of native pLF in sow's milk were achieved in three of the transgenic lines obtained. Tissue- and stage-specific pLF expressions were restricted to the mammary gland of the transgenic female mice during lactation. It was further demonstrated that the growth performance of animal pups is enhanced by directly feeding the genetically engineered milk containing enriched pLF protein in transgenic mice. Furthermore, this enhanced growth performance in suckling mice was proportional to the concentration of pLF present in milk. Publication Types: Research Support, Non-U.S. Gov't PMID: 17489602 [PubMed - indexed for MEDLINE] 459: Nat Biotechnol. 2007 May;25(5):525-31. Functional foods from biotech--an unappetizing prospect? Powell K. kendall2@nasw.org In the early 1990s, functional foods promised to solve global malnutrition and put palatable options for treating ailments on grocery shelves. Since then, a meager number of products have ripened while the rest have turned sour. Publication Types: Review PMID: 17483833 [PubMed - indexed for MEDLINE] 460: Nat Biotechnol. 2007 May;25(5):507-8. Acceptance of GM food--an experiment in six countries. Knight JG, Mather DW, Holdsworth DK, Ermen DF. Publication Types: Letter Research Support, Non-U.S. Gov't PMID: 17483829 [PubMed - indexed for MEDLINE] 461: J Invertebr Pathol. 2007 Jul;95(3):175-80. Epub 2007 Mar 31. Microbial control and biotechnology research on Bacillus thuringiensis in China. Huang DF, Zhang J, Song FP, Lang ZH. Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, China. dfhuang@mail.caas.net.cn The current status of production and application of biopesticides for pest control in China is briefly reviewed, with a focus on research advances in microbial control with Bacillus thuringiensis (Bt). These have led to improvements in Bt production, exploitation of Bt gene resources, and development of engineered Bt insecticides and transgenic Bt crops that have expanded host ranges and increased efficacy against target pests. Both conventional and biotechnology approaches need to be employed to achieve further progress in discovery, production technology, formulation processing, development of quality standards and recommended use patterns. Publication Types: Research Support, Non-U.S. Gov't PMID: 17481651 [PubMed - indexed for MEDLINE] 462: J AOAC Int. 2007 Mar-Apr;90(2):582-6. Calculation of measurement uncertainty in quantitative analysis of genetically modified organisms using intermediate precision--a practical approach. Zel J, Gruden K, Cankar K, Stebih D, Blejec A. National Institute of Biology, Vecna pot 111, 1000 Ljubljana, Slovenia. jana.zel@nib.si Quantitative characterization of nucleic acids is becoming a frequently used method in routine analysis of biological samples, one use being the detection of genetically modified organisms (GMOs). Measurement uncertainty is an important factor to be considered in these analyses, especially where precise thresholds are set in regulations. Intermediate precision, defined as a measure between repeatability and reproducibility, is a parameter describing the real situation in laboratories dealing with quantitative aspects of molecular biology methods. In this paper, we describe the top-down approach to calculating measurement uncertainty, using intermediate precision, in routine GMO testing of food and feed samples. We illustrate its practicability in defining compliance of results with regulations. The method described is also applicable to other molecular methods for a variety of laboratory diagnostics where quantitative characterization of nucleic acids is needed. Publication Types: Research Support, Non-U.S. Gov't PMID: 17474528 [PubMed - indexed for MEDLINE] 463: J Anim Sci. 2007 Aug;85(8):1946-52. Epub 2007 Apr 27. Corn expressing an Escherichia coli-derived phytase gene: a proof-of-concept nutritional study in pigs. Nyannor EK, Williams P, Bedford MR, Adeola O. Department of Animal Science, Purdue University, West Lafayette, IN 47907-2054, USA. Two experiments were conducted to investigate the concept that the addition of corn expressing an Escherichia coli-derived gene (corn-based phytase; CBP) to a P-deficient diet would improve growth performance and P utilization in pigs. An E. coli-derived microbial phytase (expressed in Pichia pastoris) sprayed onto a wheat carrier (Quantum) was included for comparison. In Exp. 1, forty-eight 10-kg pigs were blocked by BW into 6 blocks and allotted to 8 dietary treatments such that the BW among dietary treatments was similar and given free access to feed for 28 d. The dietary treatments were a negative control (NC) with no inorganic P supplementation; NC + 2, 4, or 6 g of monosodium phosphate/kg; NC + 16,500, 33,000, or 49,500 phytase units (FTU) of CBP/kg; and NC + 16,500 FTU of Quantum/kg. In Exp. 2, twenty-four 13-kg barrows were assigned to the NC, NC + 16,500 or 33,000 FTU of CBP/kg, or NC + 16,500 FTU of Quantum/kg, in a nutrient- and energy-balance study consisting of 5 d of adjustment and 5-d collection periods. The total collection method was used to determine nutrient and energy balance. Addition of CBP to the low-P NC diet linearly increased (P < 0.01) ADG, G:F, and plasma P concentration of pigs during the 28-d study. There was no difference in ADG, G:F, or plasma P concentration between pigs fed the CBP or Quantum phytase at 16,500 FTU/kg. Weight gain, G:F, and plasma P concentration of pigs increased (P < 0.01) with monosodium phosphate supplementation, confirming P deficiency of the NC diet. Linear improvements (P < 0.05) in DM digestibility and energy retention were observed with CBP supplementation of the NC diet. Although there were linear (P < 0.01) and quadratic (P < 0.05) increases in N digestibility, N retention was unaffected by CBP supplementation of the NC diet in growing pigs. Phosphorus and Ca digestibilities and retentions improved linearly and quadratically (P < 0.01) with the addition of CBP to the NC diet. There was no difference in digestive utilization of P or Ca between pigs fed CBP and Quantum phytase at 16,500 FTU/kg. The data showed that the addition of a corn expressing an E. coli-derived gene to a P-deficient diet improved growth performance and indices of P utilization in pigs, and corn expressing phytase was as efficacious as Quantum phytase when supplemented in P-deficient diets for weanling pigs. Publication Types: Comparative Study PMID: 17468432 [PubMed - indexed for MEDLINE] 464: Asia Pac J Clin Nutr. 2007;16(2):375-80. Attitudes of agricultural scientists in Indonesia towards genetically modified foods. Februhartanty J, Widyastuti TN, Iswarawanti DN. SEAMEO-TROPMED, RCCN, University of Indonesia, Campus of UI Salemba, Salemba Raya no. 6, Jakarta 10430, Indonesia. jfebruhartanty@seameo-rccn.org Conflicting arguments and partial truths on genetically modified (GM) foods have left confusion. Although studies of consumer acceptance of GM foods are numerous, the study of scientists is limited. Therefore, the main objective of this study was to assess the attitudes of scientists towards GM foods. The study was a cross sectional study. A total of 400 scientists (involved in at least one of teaching, research and consultancy) in the Bogor Agricultural Institute, Indonesia were selected randomly from its faculties of agriculture, veterinary, fishery, animal husbandry, forestry, agricultural technology, mathematics and science, and the post graduate department. Data collection was done by face-to-face interview using a structured questionnaire and self-administered questionnaire. The result showed that the majority (72.8%) of the respondents were favorably disposed towards GM foods, 14.8% were neutral, and only 12.5% were against them. The majority (78.3%) stated that they would try GM food if offered. Most (71%) reported that they were aware of the term "GM foods". Only half of the respondents felt that they had a basic understanding about GM foods. However, based on a knowledge test, 69.8% had a good knowledge score. Nearly 50% indicated that they were more exposed to news which supported GM foods. Over 90% said that there should be some form of labeling to distinguish food containing GM ingredients from non-GM foods. Attitudes were significantly associated with willingness to try GM foods if offered, restrictions on GM foods, and exposure to media reports about the pros and cons of GM foods. PMID: 17468097 [PubMed - indexed for MEDLINE] 465: Crit Rev Food Sci Nutr. 2007;47(4):363-87. Application of Failure Mode and Effect Analysis (FMEA), cause and effect analysis, and Pareto diagram in conjunction with HACCP to a corn curl manufacturing plant. Varzakas TH, Arvanitoyannis IS. Technological Educational Institute of Kalamata, School of Agricultural Sciences, Department of Processing of Agricultural Products, Hellas, Greece. The Failure Mode and Effect Analysis (FMEA) model has been applied for the risk assessment of corn curl manufacturing. A tentative approach of FMEA application to the snacks industry was attempted in an effort to exclude the presence of GMOs in the final product. This is of crucial importance both from the ethics and the legislation (Regulations EC 1829/2003; EC 1830/2003; Directive EC 18/2001) point of view. The Preliminary Hazard Analysis and the Fault Tree Analysis were used to analyze and predict the occurring failure modes in a food chain system (corn curls processing plant), based on the functions, characteristics, and/or interactions of the ingredients or the processes, upon which the system depends. Critical Control points have been identified and implemented in the cause and effect diagram (also known as Ishikawa, tree diagram, and the fishbone diagram). Finally, Pareto diagrams were employed towards the optimization of GMOs detection potential of FMEA. Publication Types: Case Reports Review PMID: 17457722 [PubMed - indexed for MEDLINE] 466: Crit Rev Food Sci Nutr. 2007;47(4):335-61. The politics and science behind GMO acceptance. Varzakas TH, Arvanitoyannis IS, Baltas H. T. H. Varzakas Technological Educational Institute of Kalamata, School of Agricultural Sciences, Department of Processing of Agricultural Products, Hellas, Greece. The question of nutritional quality has arisen in the International Community over the last few years along with other important issues such as population aging, multipopulation societies, and political conflicts. The nutritional issue is questioned both quantitatively and qualitatively. It is well known that the planet faces enormous problems with food that is available. Nowadays 20% of the population consumes approximately 80% of the produced energy and natural resources. During the last 15 years, a series of food scares and crises (BSE, dioxin, foot and mouth disease, bird flu) have seriously undermined public confidence in food producers and operators and their capacity to produce safe food. As a result, food safety has become a top priority of the European legislative authorities. Genetically Modified Organisms (GMOs) is the new food safety concern which despite the intense reactions from Non Governmental Organizations and consumer organizations have entered our lives with inadequate legislative measures to protect consumers from their consumption. The GMO issue will be the issue for discussion in the long run not only for the European Community but also for the international community as far as scientific, economical, political, ideological, ethical, and human issues are concerned. These issues are discussed in this paper along with a case of study of GM fish. Publication Types: Review PMID: 17457721 [PubMed - indexed for MEDLINE] 467: Ig Sanita Pubbl. 2007 Jan-Feb;63(1):65-94. Erratum in: Ig Sanita Pubbl. 2008 Jan;64(1):26. [Aquaculture in Italy. An integrated model of product quality control] [Article in Italian] De Giusti M, Cocchieri RA, De Vito E, Grasso GM, Ortaggi G, Reali D, Ricciardi G, Romano-Spica V, Boccia A. Università degli Studi di Roma La Sapienza, Dipartimento de Medicina Sperimantale, Sezione de Medicina Clinica e Sanità Pubblica. Aquaculture is becoming increasingly diffuse even in Italy. The increased production introduces new problems such as product quality control and process safety. This article presents the results of a research project, funded by the Ministry of the Environment, whose aim was to evaluate and promote aquaculture product quality and safety in an environmentally responsible way. Four intensive land-based and offshore aquaculture sites were monitored to evaluate microbiological, biological and chemical (i.e. polychlorinated biphenyls and endocrine disruptors) quality of water, products and fish feed. In total 154 samples were analysed, of which 66 were water samples, 55 product samples and 33 feed samples. Salmonella and other enteric pathogens were absent in products and the aquatic environment, while other environmental pathogens of the Vibrio species were detected. Bacterial load and fecal indicators were found to be higher in off-shore products and in mussels from all aquaculture sites. PCBs were detected in all products in concentrations below 2 microg/g fresh product (Food and Drug Administration), but on average, higher concentrations were detected in off-shore products. No estrogen mimetic activity was detected in fish feed, in contrast it was detected in offshore products and water. Product quality was found to be strictly correlated with the quality of the environment. Genetically modified organisms were detected in fish feed but no integration of genetic material in products occurred. Publication Types: English Abstract PMID: 17450652 [PubMed - indexed for MEDLINE] 468: Environ Biosafety Res. 2006 Jul-Sep;5(3):151-68. Epub 2007 Mar 17. Meteorological input data requirements to predict cross-pollination of GMO maize with Lagrangian approaches. Lipsius K, Wilhelm R, Richter O, Schmalstieg KJ, Schiemann J. Institute for Geoecology, Environmental Systems Analysis Group, Technical University Braunschweig, Germany. K.Lipsius@tu-bs.de Modeling pollen dispersal to predict cross-pollination is of great importance for the ongoing discussion of adventitious presence of genetically modified material in food and feed. Two different modeling approaches for pollen dispersal were used to simulate two years of data for the rate of cross-pollination of non-GM maize (Zea mays (L.)) fields by pollen from a central 1 ha transgenic field. The models combine the processes of wind pollen dispersal (transport) and pollen competition. Both models used for the simulation of pollen dispersal were Lagrangian approaches: a stochastic particle Lagrange model and a Lagrangian transfer function model. Both modeling approaches proved to be appropriate for the simulation of the cross-pollination rates. However, model performance differed significantly between years. We considered different complexity in meteorological input data. Predictions compare well with experimental results for all simplification steps, except that systematic deviations occurred when only main wind direction was used. Concluding, it can be pointed out that both models might be adapted to other pollen dispersal experiments of different crops and plot sizes, when wind direction statistics are available. However, calibration of certain model parameters is necessary. Publication Types: Comparative Study Research Support, Non-U.S. Gov't PMID: 17445511 [PubMed - indexed for MEDLINE] 469: Environ Biosafety Res. 2006 Jul-Sep;5(3):127-49. Epub 2007 Mar 24. The interplay between societal concerns and the regulatory frame on GM crops in the European Union. Devos Y, Reheul D, De Waele D, Van Speybroeck L. Department of Plant Production, Faculty of Bioscience Engineering, Ghent University, Coupure Links 653, 9000 Ghent, Belgium. Yann.Devos@UGent.be Recapitulating how genetic modification technology and its agro-food products aroused strong societal opposition in the European Union, this paper demonstrates how this opposition contributed to shape the European regulatory frame on GM crops. More specifically, it describes how this opposition contributed to a de facto moratorium on the commercialization of new GM crop events in the end of the nineties. From this period onwards, the regulatory frame has been continuously revised in order to slow down further erosion of public and market confidence. Various scientific and technical reforms were made to meet societal concerns relating to the safety of GM crops. In this context, the precautionary principle, environmental post-market monitoring and traceability were adopted as ways to cope with scientific uncertainties. Labeling, traceability, co-existence and public information were installed in an attempt to meet the general public request for more information about GM agro-food products, and the specific demand to respect the consumers' and farmers' freedom of choice. Despite these efforts, today, the explicit role of public participation and/or ethical consultation during authorization procedures is at best minimal. Moreover, no legal room was created to progress to an integral sustainability evaluation during market procedures. It remains to be seen whether the recent policy shift towards greater transparency about value judgments, plural viewpoints and scientific uncertainties will be one step forward in integrating ethical concerns more explicitly in risk analysis. As such, the regulatory frame stands open for further interpretation, reflecting in various degrees a continued interplay with societal concerns relating to GM agro-food products. In this regard, both societal concerns and diversely interpreted regulatory criteria can be inferred as signaling a request - and even a quest - to render more explicit the broader-than-scientific dimension of the actual risk analysis. PMID: 17445510 [PubMed - indexed for MEDLINE] 470: J Sep Sci. 2007 Mar;30(4):579-85. A simple capillary gel electrophoresis approach for efficient and reproducible DNA separations. Analysis of genetically modified soy and maize. Sánchez L, González R, Crego AL, Cifuentes A. Institute of Industrial Fermentations (CSIC), Juan de la Cierva 3, Madrid, Spain. It is generally assumed that in order to achieve suitable separations of DNA fragments, capillary gel electrophoresis (CGE)-coated capillaries should be used. In this work, a new method is presented that allows to obtain reproducible CGE separations of DNA fragments using bare fused-silica capillaries without any previous coating step. The proposed method only requires: (i) a capillary washing with 0.1 M hydrochloric acid between injections and (ii) a running buffer composed of Tris-phosphate-ethylenediamine tetraacetic acid (EDTA) and 4.5% of 2-hydroxyethyl cellulose (HEC) as sieving polymer. The use of this new CGE procedure gives highly resolved and reproducible separations of DNA fragments ranging from 50 to 750 bp. The separation of these DNA fragments is accomplished in less than 30 min with efficiencies up to 1.7 x 10(6) plates/m. Reproducibility values of migration times (given as %RSD) for the analyzed DNA fragments are better than 1.0% (n = 4) for the same day, 2.2% (n = 16) for four different days, and 2.3% (n = 16) for four different capillaries. The usefulness of this separation method is demonstrated by detecting genetically modified maize and genetically modified soy after DNA amplification by PCR. This new CGE procedure together with LIF as detector provides sensitive analysis of 0.9% of Bt11 maize, Mon810 maize, and Roundup Ready soy in flours with S/ N up to 542. These results demonstrate the usefulness of this procedure to fulfill the European regulation on detection of genetically modified organisms in foods. Publication Types: Research Support, Non-U.S. Gov't PMID: 17444227 [PubMed - indexed for MEDLINE] 471: Dev Neurobiol. 2007 Feb 1;67(2):189-204. The serotonin receptor SER-1 (5HT2ce) contributes to the regulation of locomotion in Caenorhabditis elegans. Dernovici S, Starc T, Dent JA, Ribeiro P. Institute of Parasitology, McGill University, Macdonald Campus, Ste. Anne de Bellevue, Quebec, Canada H9X 3V9. Serotonin (5-hydroxytryptamine: 5HT) is an important neuroactive substance in the model roundworm, Caenorhabditis elegans. Aside from having effects in feeding and egg-laying, 5HT inhibits motility and also modulates several locomotory behaviors, notably food-induced slowing and foraging. Recent evidence showed that a serotonergic 5HT2-like receptor named SER-1 (also known as 5HT2ce) was responsible for the effect of 5HT on egg-laying. Here we confirm this observation and show that SER-1 also plays an important role in locomotion. A mutant lacking SER-1 was found to be highly resistant to exogenous 5HT in the absence of food and this resistant phenotype was rescued by reintroducing the SER-1 gene in a mutant background. Pharmacological studies showed that the same antagonists that blocked the activity of recombinant SER-1 in vitro also inhibited the effect of 5HT on motility, suggesting the same receptor was responsible for both effects. When tested for locomotory behaviors, the SER-1 mutant was found to be moderately defective in food-induced slowing. In addition, the mutant changed direction more frequently than the wildtype when searching for food, suggesting that SER-1 may play a role in navigational control during foraging. Both these effects required the presence of MOD-1, a 5HT gated chloride channel, and the results indicate that SER-1 and MOD-1 modulate these behaviors through a common pathway. On the basis of expression analysis of a ser-1::GFP translational fusion, SER-1 is prominently located in central, integrating neurons of the head ganglia (RIA and RIC) but not the body wall musculature. The evidence suggests that SER-1 controls locomotion through indirect modulation of neuromuscular circuits and has effects both on speed and direction of movement. (c) 2006 Wiley Periodicals, Inc. Publication Types: Research Support, Non-U.S. Gov't PMID: 17443782 [PubMed - indexed for MEDLINE] 472: Transgenic Res. 2007 Jun;16(3):261-80. Epub 2007 Apr 14. Biosafety and risk assessment framework for selectable marker genes in transgenic crop plants: a case of the science not supporting the politics. Ramessar K, Peremarti A, Gómez-Galera S, Naqvi S, Moralejo M, Muñoz P, Capell T, Christou P. Departament de Produccio Vegetal i Ciencia Forestal, Universitat de Lleida, Av. Alcalde Rovira Roure, 191, Lleida 25198, Spain. Selectable marker gene systems are vital for the development of transgenic crops. Since the creation of the first transgenic plants in the early 1980s and their subsequent commercialization worldwide over almost an entire decade, antibiotic and herbicide resistance selectable marker gene systems have been an integral feature of plant genetic modification. Without them, creating transgenic crops is not feasible on purely economic and practical terms. These systems allow the relatively straightforward identification and selection of plants that have stably incorporated not only the marker genes but also genes of interest, for example herbicide tolerance and pest resistance. Bacterial antibiotic resistance genes are also crucial in molecular biology manipulations in the laboratory. An unprecedented debate has accompanied the development and commercialization of transgenic crops. Divergent policies and their implementation in the European Union on one hand and the rest of the world on the other (industrialized and developing countries alike), have resulted in disputes with serious consequences on agricultural policy, world trade and food security. A lot of research effort has been directed towards the development of marker-free transformation or systems to remove selectable markers. Such research has been in a large part motivated by perceived problems with antibiotic resistance selectable markers; however, it is not justified from a safety point of view. The aim of this review is to discuss in some detail the currently available scientific evidence that overwhelmingly argues for the safety of these marker gene systems. Our conclusion, supported by numerous studies, most of which are commissioned by some of the very parties that have taken a position against the use of antibiotic selectable marker gene systems, is that there is no scientific basis to argue against the use and presence of selectable marker genes as a class in transgenic plants. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 17436060 [PubMed - indexed for MEDLINE] 473: Trends Biotechnol. 2007 Jun;25(6):239-41. Epub 2007 Apr 12. Reduced terpene levels in cottonseed add food to fiber. Townsend BJ, Llewellyn DJ. CSIRO Plant Industry, GPO Box 1600, Canberra, ACT 2601, Australia. belinda.townsend@bbsrc.ac.uk Using RNA interference (RNAi) technology, the levels of a toxic phytoprotectant have recently been reduced specifically in the seeds of cotton to generate a novel dual-purpose crop. By engineering an endogenous terpene pathway, there is now the exciting potential for an added-value, genetically modified crop with the cash value of the fiber supported by the improved nutritional value and expanded food and feed use for the cottonseed, which is normally a low-value by-product. Publication Types: Review PMID: 17433845 [PubMed - indexed for MEDLINE] 474: Autophagy. 2007 Jul-Aug;3(4):360-2. Epub 2007 Jul 19. Comment on: Plant Cell Physiol. 2006 Dec;47(12):1641-52. Constitutive autophagy in plant root cells. Yano K, Suzuki T, Moriyasu Y. Graduate School of Nutritional and Environmental Sciences, University of Shizuoka, Shizuoka, Japan. In previous studies, using a membrane-permeable protease inhibitor, E-64d, we showed that autophagy occurs constitutively in the root cells of barley and Arabidopsis. In the present study, a fusion protein composed of the autophagy-related protein AtAtg8 and green fluorescent protein (GFP) was expressed in Arabidopsis to visualize autophagosomes. We first confirmed the presence of autophagosomes with GFP fluorescence in the root cells of seedlings grown on a nutrient-sufficient medium. The number of autophagosomes changed as the root cells grew and differentiated. In cells near the apical meristem, autophagosomes were scarcely found. However, a small but significant number of autophagosomes existed in the elongation zone. More autophagosomes were found in the differentiation zone where cell growth ceases but the cells start to form root hair. In addition, we confirmed that autophagy is activated under starvation conditions in Arabidopsis root cells. When the root tips were cultured in a sucrose-free medium, the number of autophagosomes increased in the elongation and differentiation zones, and a significant number of autophagosomes appeared in cells near the apical meristem. The results suggest that autophagy in plant root cells is involved not only in nutrient recycling under nutrient-limiting conditions but also in cell growth and root hair formation. Publication Types: Comment PMID: 17426438 [PubMed - indexed for MEDLINE] 475: Wei Sheng Yan Jiu. 2007 Jan;36(1):45-8. [Expression and identification of Cry 1Ie in Bacillus coli] [Article in Chinese] Xu H, Zhang X, Li F, Wang G. Institute of Nutrition and Food Safety, Chinese Center for Disease Control and Prevention, Beijing. OBJECTIVE: In this study, Cry1Ie expressed protein in the transgenic Cry1Ie maize was used as a model protein to establish a technical platform for the safety assessment of other genetically modified plants. METHODS: Bacillus coli expression system was used to express Cry1Ie protein and then the expressed protein was purified by SDS-PAGE. The equivalence of Cry1Ie proteins expressed in Bacillus coli and in transgenic Cry1Ie maize was analyzed by their immunorecognition and bioactivity. RESULTS: Cry1Ie protein was highly expressed in Bacillus coli and could be segregated and purified by SDS-PAGE. Cry1Ie expressed protein in Bacillus coli is substantially equivalent to that of maize genetically modified with Cry1Ie in tests of immunorecognition western blot and bioactivity in target pest. CONCLUSION: The established technical platform of external Cry1Ie expressed protein can be applied to assess the food safety of genetically modified plants. Publication Types: English Abstract PMID: 17424848 [PubMed - in process] 476: Rev Med Univ Navarra. 2006 Oct-Dec;50(4):62-70. [New challenges in basic and applied nutrition] [Article in Spanish] Palou A. Laboratorio de Biología Molecular, Nutrición y Biotecnología (Nutrigenómica), Universitat de les Illes Balears, Palma de Mallorca. The quality standards for nutrition and food have undergone major changes during recent years in parallel with the increase in scientific knowledge in the area of food and health. Trends in consumer demands are changing accordingly. While during the last century our concerns were firstly focused on ensuring the availability of basic foods and later on ensuring its safety, nowadays our society is concentrating on improving wellbeing and, particularly, on tackling and preventing the major chronic diseases such as cardiovascular diseases, diabetes, obesity, diverse types of cancer, osteoporosis and autoimmune diseases. These problems account for almost 50% of the diseases and 60% of deaths over the world, and are directly related with the food we eat, irrespective of the importance of other factors (age, sex, physical exercise, genetic predisposition, nicotine poisoning or alcoholism). Europe has decided to go ahead with a major legislative change in the food sector (the Regulation on nutrition and health claims made on foods) which opens new perspectives to improve public health and offer clear economic growth for the health-related food sector. Putting into practice this new legislation and its consequences constitutes a major challenge. Nevertheless, new challenges are foreseen: our response to food depends on our individual genetic characteristics (nutrigenetics); it also depends on the individual history that is being imprinted (in a permanent or temporary form) on our chromosomes (epigenetics), as determined by the individual life style (food, different episodes and facts, including emotions) and, particularly, nutrition during the most active stages of our development. For the future, the additional contribution we will ask of food is to promote health and well-being in all facets. The requirement is that it should develop in a free framework, based on the best scientific available advice, with transparency as a fundamental guarantee. In this context, new disciplines (e.g. nutrigenomics) are emerging and new trends towards a more conscious, better informed, consumer are foreseen that will allow an appropriate reaction to these new challenges in the medium and long term. Publication Types: English Abstract Review PMID: 17424771 [PubMed - indexed for MEDLINE] 477: Anal Chim Acta. 2007 Apr 25;589(2):159-65. Epub 2007 Feb 25. Solubilisation and binding characteristics of a recombinant beta2-adrenergic receptor expressed in the membrane of Escherichia coli for the multianalyte detection of beta-agonists and antagonists residues in food-producing animals. Danyi S, Degand G, Duez C, Granier B, Maghuin-Rogister G, Scippo ML. Department of Food Sciences, Laboratory of Analysis, Faculty of Veterinary Medicine, University of Liège, Sart-Tilman, Liège, Belgium. sdanyi@ulg.ac.be The number of substances with beta-agonistic activity, illegally introduced in meat production or in sports doping as anabolic or beta-blocking agents is increasing. Analytical methods suited for their multianalyte detection are thus necessary. In this perspective, receptor assays were developed. The research activities undertaken in this study describe the solubilisation of a recombinant human beta(2)-adrenergic receptor produced in the inner membrane of genetically modified Escherichia coli, using the detergent n-dodecyl-beta-d-maltoside. Its potential to detect the presence of beta-agonists or beta-blockers in biological samples was evaluated. The solubilised beta(2)-adrenergic receptor retained its binding affinity in a radio-receptor assay based on the competition for the binding to receptors between a ligand (beta-agonist or antagonist) and the radioligand [(125)I]iodocyanopindolol. The IC(50) values ranged from 5+/-1 x 10(-8) M (clenbuterol) to 8+/-2 x 10(-6) M (isoxsuprine) for the beta-agonists tested and from 1.5+/-0.2 x 10(-10) M (carazolol) to 1.2+/-0.2 x 10(-5) M (metoprolol) for the beta-blockers tested. It was shown to have a lower limit of detection than a radio-receptor assay using the solubilised beta(2)-adrenoceptor expressed in a mammalian cell line. The solubilised recombinant human beta(2)-adrenoreceptor expressed in E. coli would be a useful tool to develop non radioactive multianalyte screening methods. Publication Types: Research Support, Non-U.S. Gov't PMID: 17418176 [PubMed - indexed for MEDLINE] 478: Transgenic Res. 2007 Dec;16(6):795-812. Epub 2007 Apr 6. Bitrophic and tritrophic effects of Bt Cry3A transgenic potato on beneficial, non-target, beetles. Ferry N, Mulligan EA, Majerus ME, Gatehouse AM. School of Biology, Institute for Research on Environment and Sustainability, University of Newcastle Upon Tyne, Devonshire Building, Newcastle NE1 7RU, UK. Insect-resistant transgenic plants have been suggested to have unpredictable effects on the biodiversity of the agro-ecosystem, including potential effects on insect natural enemies, beneficial in control of crop pests. Whilst carnivorous as adults, many of these predators may also consume plant tissues, in particular plant pollen and nectar. Coleoptera are important in terms of agro-ecological research not only because of the large number of species in this order, but also because of their role as biological control agents. Thus any detrimental impact on this group of insects would be highly undesirable. The effects of potato expressing the coleopteran-specific Bacillus thuringiensis delta-endotoxin Cry3A (Bt Cry3A) on the ladybird beetle Harmonia axyridis and the carabid beetle Nebria brevicollis were investigated via the bitrophic interaction of the adult ladybird with potato flowers and the tritrophic interaction of the carabid consuming a non-target potato pest. Immunoassays confirmed accumulation of the transgene product in potato leaves and floral tissues (at levels of up to 0.01% (pollen) and 0.0285% (anthers) of total soluble protein). Despite H. axyridis and N. brevicollis belonging to the targeted insect order, no significant effects upon survival or overall body mass change of either beetle were observed. Furthermore, Bt Cry3A had no detrimental effects on reproductive fitness of either beetle species, either in terms of fecundity or subsequent egg viability. Behavioural analysis revealed no significant impact of Bt Cry3A on beetle activity or locomoter behaviour. Ligand blots indicate that this is due to either the absence of Bt-binding sites in brush border membrane vesicles (BBMV) isolated from Nebria brevicollis, or in the case of Harmonia axyridis, the binding did not functionally lead to behavioural or physical effects. Publication Types: Comparative Study PMID: 17415673 [PubMed - indexed for MEDLINE] 479: Bull Entomol Res. 2007 Apr;97(2):211-5. Evaluation of Bt-toxin uptake by the non-target herbivore, Myzus persicae (Hemiptera: Aphididae), feeding on transgenic oilseed rape. Burgio G, Lanzoni A, Accinelli G, Dinelli G, Bonetti A, Marotti I, Ramilli F. Dipartimento di Scienze e Tecnologie Agroambientali, Alma Mater Studiorum-Università di Bologna, Viale Fanin 42, 40127 Bologna, Italy. gburgio@entom.agrsci.unibo.it As consequence of the concern about the biosafety of genetically modified plants, biological and ecological studies are considered crucial for environmental risk assessment. Laboratory experiments were carried out in order to evaluate the transfer of the Cry1Ac Bt-toxin from a transgenic Bt-oilseed rape to a non-target pest, Myzus persicae Sulzer. Cry1Ac protein levels in plants and aphids were determined using a double sandwich enzyme-linked immunosorbent assay. Phloem sap from (Bt+) and (Bt-) oilseed rape plants was collected from leaves using a standard method of extraction in an EDTA buffer. Bt-toxin was present in phloem sap, with a mean concentration of 2.7 +/- 1.46 ppb, corresponding to a 24-fold lower level than in oilseed rape leaves. Toxin was also detected in aphid samples, with a mean concentration in the positive samples of 2.0 +/- 0.8 ppb. The evidence that Bt-toxin remains in herbivores, in this case an aphid, could be useful to clarify functional aspects linked to possible consequences of Bt-crops on food chains involving herbivore-natural enemy trophic systems. Further studies are needed in order to improve the knowledge on the functional aspects linked to the transfer of the Cry1Ac Bt-toxin from GM-oilseed rape to aphids and their possible consequence. Publication Types: Research Support, Non-U.S. Gov't PMID: 17411484 [PubMed - indexed for MEDLINE] 480: Annu Rev Phytopathol. 2007;45:173-202. Safety of virus-resistant transgenic plants two decades after their introduction: lessons from realistic field risk assessment studies. Fuchs M, Gonsalves D. Department of Plant Pathology, Cornell University, New York State Agricultural Experiment Station, Geneva, NY 14456, USA. mf13@cornell.edu Potential safety issues have been raised with the development and release of virus-resistant transgenic plants. This review focuses on safety assessment with a special emphasis on crops that have been commercialized or extensively tested in the field such as squash, papaya, plum, grape, and sugar beet. We discuss topics commonly perceived to be of concern to the environment and to human health--heteroencapsidation, recombination, synergism, gene flow, impact on nontarget organisms, and food safety in terms of allergenicity. The wealth of field observations and experimental data is critically evaluated to draw inferences on the most relevant issues. We also express inside views on the safety and benefits of virus-resistant transgenic plants, and recommend realistic risk assessment approaches to assist their timely deregulation and release. Publication Types: Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Review PMID: 17408355 [PubMed - indexed for MEDLINE] 481: J Agric Food Chem. 2007 May 2;55(9):3268-74. Epub 2007 Apr 4. Toward metrological traceability for DNA fragment ratios in GM quantification. 3. Suitability of DNA calibrants studied with a MON 810 corn model. Charels D, Broeders S, Corbisier P, Trapmann S, Schimmel H, Emons H. European Commission, Joint Research Centre, Institute for Reference Materials and Measurements, IRMM, Retieseweg 111, 2440 Geel, Belgium. diana.charels@ec.europa.eu The quantification of GMOs by real-time PCR relies on an external calibrant. In this paper the suitability of two DNA calibrants, genomic DNA from plant leaves and plasmidic DNA, was investigated. The PCR efficiencies, the correlation coefficients of the calibration curves, and the ratios between PCR efficiencies of transgenic and endogenous sequences were compared for both calibrants using 59 data sets produced by 43 laboratories. There were no significant differences between plasmidic and genomic DNA except for the PCR efficiencies of the calibration curves for the transgene of the construct-specific real-time PCR method. In the GM system investigated, PCR efficiencies of plasmidic calibrants were slightly closer to the PCR efficiencies observed for the unknowns than those of the genomic DNA calibrant. Therefore, plasmidic DNA was the more suitable calibrant for the PCR measurements on genomic DNA extracted from MON 810 seeds. It is shown that plasmidic DNA is an appropriate choice for the calibration of measurements of MON 810 corn with respect to the DNA copy number ratio. PMID: 17407307 [PubMed - indexed for MEDLINE] 482: J Agric Food Chem. 2007 May 2;55(9):3258-67. Epub 2007 Apr 4. Toward metrological traceability for DNA fragment ratios in GM quantification. 2. Systematic study of parameters influencing the quantitative determination of MON 810 corn by real-time PCR. Charels D, Broeders S, Corbisier P, Trapmann S, Schimmel H, Linsinger T, Emons H. European Commission, Joint Research Centre, Institute for Reference Materials and Measurements (IRMM), Retieseweg 111, 2440 Geel, Belgium. diana.charels@ec.europa.eu This paper is part of a set of three papers investigating metrological traceability of the quantification of DNA fragments as, for instance, used for quantification of genetic modifications. This paper evaluates the possible impact of several factors on results of real-time Polymerase Chain Reaction (PCR) measurements. It was found that the particle size of the powder samples does not have an influence, whereas the nature of the calibrant (plasmidic or genomic DNA) has a significant effect. Moreover, two real-time PCR detection methods (construct-specific and event-specific) for MON 810 corn were compared. The results obtained in a specifically designed interlaboratory study revealed a significant influence of the DNA extraction method on measurement results when the MON 810 construct-specific real-time PCR detection method was applied. Statistical analyses confirmed the importance of validating DNA extraction methods in conjunction with real-time PCR methods. PMID: 17407306 [PubMed - indexed for MEDLINE] 483: J Agric Food Chem. 2007 May 2;55(9):3249-57. Epub 2007 Apr 4. Toward metrological traceability for DNA fragment ratios in GM quantification. 1. Effect of DNA extraction methods on the quantitative determination of Bt176 corn by real-time PCR. Corbisier P, Broothaerts W, Gioria S, Schimmel H, Burns M, Baoutina A, Emslie KR, Furui S, Kurosawa Y, Holden MJ, Kim HH, Lee YM, Kawaharasaki M, Sin D, Wang J. European Commission, Joint Research Centre, Institute for Reference Materials and Measurements (IRMM), Retieseweg 111, 2440 Geel, Belgium. philippe.corbisier@ec.europa.edu. An international CCQM-P60 pilot study involving eight national metrological institutes was organized to investigate if the quantification of genetically modified (GM) corn powder by real-time PCR was affected by the DNA extraction method applied. Four commonly used extraction methods were compared for the extraction of DNA from a GM Bt176 corn powder. The CTAB-based method yielded the highest DNA template quantity and quality. A difference in the 260 nm/230 nm absorbance ratio was observed among the different extraction methods. Real-time amplification of sequences specific for endogenous genes zein and hmg as well as transgenic sequences within the cryIA(b) gene and a fragment covering the junction between the transformed DNA and the plant genome were used to determine the GM percentage. The detection of the transgenic gene was affected by the quantity and quality of template used for the PCR reaction. The Bt176 percentages measured on diluted or purified templates were statistically different depending on the extraction method applied. PMID: 17407305 [PubMed - indexed for MEDLINE] 484: Nat Protoc. 2006;1(6):2828-30. Assessing hoarding in mice. Deacon RM. Department of Experimental Psychology, University of Oxford, South Parks Road, Oxford OX1 3UD, UK. robert.deacon@psy.ox.ac.uk Hoarding is a species-typical behavior shown by rodents, as well as other animals. By hoarding, the rodent secures a food supply for times of emergency (for example, when threatened by a predator) or for times of seasonal adversity such as winter. Scatter hoarding, as seen typically in squirrels and birds, involves placing small caches of food in hidden places, generally underground. Most rodents, however, hoard a supply of food in or near the home base--for example, in 'larders' near the sleeping quarters in a burrow. In the laboratory, measurement of hoarding involves simply weighing the food transported into the home cage from an external source, but the route to that source must be secure and animal-proof; for example, there should be no holes large enough to permit escape of a mouse, and no weak points that could be enlarged by gnawing. A suitable and easily constructed apparatus is described in the protocol. Hoarding has been shown to be sensitive to brain lesions and pharmacological agents, and is a suitable test for species-typical behavior in genetically modified mice. Publication Types: Research Support, Non-U.S. Gov't PMID: 17406541 [PubMed - indexed for MEDLINE] 485: Nat Protoc. 2006;1(1):122-4. Digging and marble burying in mice: simple methods for in vivo identification of biological impacts. Deacon RM. Department of Experimental Psychology, University of Oxford, South Parks Road, Oxford OX1 3UD, UK. robert.deacon@psy.ox.ac.uk Mice exhibit various species-typical behaviors such as digging and burrowing. They dig in the ground to find food, to hoard food, to create a refuge from predators or cold and to make a safe nursery area for the young. In the laboratory, mice dig vigorously in deep bedding such as wood chips. This behavior is sensitive to strain differences and drugs. For example, the effects of anxiolytics and 5-HT-active compounds, including those used clinically for obsessive-compulsive disorder (OCD), can be detected. Digging can be quantified by manual timing. Alternatively, the bedding can be covered with glass marbles and the number buried can be counted after a set time. These behaviors can be assessed using very little specialized equipment, and results can be obtained from ten animals in about an hour. Species-typical behaviors may be sensitive to a wide variety of treatments, and their simplicity and ability to yield robust quantitative data might be particularly useful in assessing genetically modified mice, even in laboratories not primarily oriented to behavioral work. Publication Types: Research Support, Non-U.S. Gov't PMID: 17406223 [PubMed - indexed for MEDLINE] 486: J Agric Food Chem. 2007 May 2;55(9):3351-7. Epub 2007 Apr 3. Detection methods for biotech cotton MON 15985 and MON 88913 by PCR. Lee SH, Kim JK, Yi BY. Gene Analysis Laboratory, Experiment Research Institute of National Agricultural Products Quality Management Service, Seoul 150-043, South Korea. starlee@naqs.go.kr Plants derived through agricultural biotechnology, or genetically modified organisms (GMOs), may affect human health and ecological environment. A living GMO is also called a living modified organism (LMO). Biotech cotton is a GMO in food or feed and also an LMO in the environment. Recently, two varieties of biotech cotton, MON 15985 and MON 88913, were developed by Monsanto Co. The detection method is an essential element for the GMO labeling system or LMO management of biotech plants. In this paper, two primer pairs and probes were designed for specific amplification of 116 and 120 bp PCR products from MON 15985 and MON 88913, respectively, with no amplification from any other biotech cotton. Limits of detection of the qualitative method were all 0.05% for MON 15985 and MON 88913. The quantitative method was developed using a TaqMan real-time PCR. A synthetic plasmid, as a reference molecule, was constructed from a taxon-specific DNA sequence of cotton and two construct-specific DNA sequences of MON 15985 and MON 88913. The quantitative method was validated using six samples that contained levels of biotech cotton mixed with conventional cotton ranging from 0.1 to 10.0%. As a result, the biases from the true value and the relative deviations were all within the range of +/-20%. Limits of quantitation of the quantitative method were all 0.1%. Consequently, it is reported that the proposed detection methods were applicable for qualitative and quantitative analyses for biotech cotton MON 15985 and MON 88913. PMID: 17402745 [PubMed - indexed for MEDLINE] 487: EMBO Rep. 2007 Apr;8(4):305-8. Comment in: EMBO Rep. 2007 Apr;8(4):309-15. EMBO Rep. 2007 Jul;8(7):612-3. The precautionary principle should not be used as a basis for decision-making. Talking point on the precautionary principle. Peterson M. Department of History and Philosophy of Science, University of Cambridge, UK. mbp24@cam.ac.uk PMID: 17401402 [PubMed - indexed for MEDLINE] 488: J Fish Dis. 2007 Apr;30(4):201-12. Evaluation of stress- and immune-response biomarkers in Atlantic salmon, Salmo salar L., fed different levels of genetically modified maize (Bt maize), compared with its near-isogenic parental line and a commercial suprex maize. Sagstad A, Sanden M, Haugland Ø, Hansen AC, Olsvik PA, Hemre GI. National Institute of Nutrition and Seafood Research, NIFES, Bergen, Norway. The present study was designed to evaluate if genetically modified (GM) maize (Bt maize, event MON810) compared with the near-isogenic non-modified (nGM) maize variety, added as a starch source at low or high inclusions, affected fish health of post-smolt Atlantic salmon, Salmo salar L. To evaluate the health impact, selected stress- and immune-response biomarkers were quantified at the gene transcript (mRNA) level, and some also at the protein level. The diets with low or high inclusions of GM maize, and its near-isogenic nGM parental line, were compared to a control diet containing GM-free suprex maize (reference diet) as the only starch source. Total superoxide dismutase (SOD) activity in liver and distal intestine was significantly higher in fish fed GM maize compared with fish fed nGM maize and with the reference diet group. Fish fed GM maize showed significantly lower catalase (CAT) activity in liver compared with fish fed nGM maize and to the reference diet group. In contrast, CAT activity in distal intestine was significantly higher for fish fed GM maize compared with fish fed reference diet. Protein level of heat shock protein 70 (HSP70) in liver was significantly higher in fish fed GM maize compared with fish fed the reference diet. No diet-related differences were found in normalized gene expression of SOD, CAT or HSP70 in liver or distal intestine. Normalized gene expression of interleukin-1 beta in spleen and head-kidney did not vary significantly between diet groups. Interestingly, fish fed high GM maize showed a significantly larger proportion of plasma granulocytes, a significantly larger sum of plasma granulocyte and monocyte proportions, but a significantly smaller proportion of plasma lymphocytes, compared with fish fed high nGM maize. In conclusion, Atlantic salmon fed GM maize showed some small changes in stress protein levels and activities, but none of these changes were comparable to the normalized gene expression levels analysed for these stress proteins. GM maize seemed to induce significant changes in white blood cell populations which are associated with an immune response. Publication Types: Comparative Study PMID: 17394522 [PubMed - indexed for MEDLINE] 489: Lipids. 2007 Apr;42(3):179-85. Epub 2007 Mar 14. Engineering oilseed plants for a sustainable, land-based source of long chain polyunsaturated fatty acids. Damude HG, Kinney AJ. Crop Genetics Research, DuPont Experimental Station, Wilmington, DE 19880-0353, USA. Numerous clinical studies have demonstrated the cardiovascular and mental health benefits of including very long chain omega-3 polyunsaturated fatty acids, namely eicospentaenoic acid (EPA) and docosohexaenoic acid (DHA) in the human diet. Certain fish oils can be a rich source of omega-3 long chain polyunsaturated fatty acids although processed marine oils are generally undesirable as food ingredients because of the associated objectionable flavors and contaminants that are difficult and cost-prohibitive to remove. Oilseed plants rich in omega-3 fatty acids, such as flax and walnut oils, contain only the 18-carbon omega-3 polyunsaturated fatty acid alpha-linolenic acid, which is poorly converted by the human body to EPA and DHA. It is now possible to engineer common omega-6 rich oilseeds such as soybean and canola to produce EPA and DHA and this has been the focus of a number of academic and industrial research groups. Recent advances and future prospects in the production of EPA and DHA in oilseed crops are discussed here. Publication Types: Review PMID: 17393224 [PubMed - indexed for MEDLINE] 490: Asia Pac J Clin Nutr. 2007;16 Suppl 1:122-6. Fruit quality of transgenic tomatoes with suppressed expression of LeETR1 and LeETR2 genes. Bao B, Ke L, Jiang J, Ying T. Department of Food Science and Nutrition, Zhejiang University, 268 Kaixuan Road, Hangzhou, Zhejiang, China 310029. Tomato fruit is renowned for its high concentration of phyto-nutrients such as lycopene and carotenoids, overall contribution to nutrition and human health. The effect of antisense suppression of ethylene receptor genes LeETR1 and LeETR2 over the quality of tomato fruit was investigated in this paper. During the different stages of ripening, the fruit of antisense transgenic tomatoes of ale1 and ale2, compared to their wild type B1, showed higher total soluble solids, acidity and electrolytes accumulations and color development; lower fruit firmness, fruit viscosity and fruit elasticity. However, no significant difference of Vc content, total sugar, fruit pH value and fruit pigments between transgenic lines and B1 were noticed. ale1 and ale2 showed shortened shelf life. The data suggest that fruit with suppressed LeETR1 and LeETR2 genes expression have stronger ethylene response, which accelerate fruit ripening and greatly altered tomato variety characteristics. Publication Types: Research Support, Non-U.S. Gov't PMID: 17392089 [PubMed - indexed for MEDLINE] 491: Am J Physiol Endocrinol Metab. 2007 Jul;293(1):E252-8. Epub 2007 Mar 27. Melanocortin activation of nucleus of the solitary tract avoids anorectic tachyphylaxis and induces prolonged weight loss. Li G, Zhang Y, Rodrigues E, Zheng D, Matheny M, Cheng KY, Scarpace PJ. Department of Pharmacology and Therapeutics, University of Florida College of Medicine, Gainesville, Florida 32610, USA. To examine the role of the brain stem melanocortin system in long-term energy regulation, we assessed the effects of overproduction of proopiomelanocortin (POMC) in the caudal brain stem of F344xBN rats with adult-onset obesity. Recombinant adeno-associated viral vector encoding POMC gene was delivered to the nucleus of solitary tract (NTS) in the hindbrain, and food intake, body weight, glucose and fat metabolism, brown adipose tissue thermogenesis, and mRNA levels of neuropeptides and melanocortin receptors were assessed. POMC delivery resulted in sustained reduction in food intake and body weight over 42 days and improved insulin sensitivity. At death, in recombinant adeno-associated viral vector-POMC-treated rats vs. control rats, alpha-melanocyte-stimulating hormone in NTS increased nearly 21-fold, whereas hypothalamic alpha-melanocyte-stimulating hormone remained unchanged. Visceral adiposity decreased by 37%; tissue triglyceride content diminished by 26% and 47% in liver and muscle, respectively; serum triglyceride and nonesterified fatty acids were reduced by 35% and 34%, respectively; phosphorylation of acetyl-CoA carboxylase was elevated by 63% in soleus muscle; brown adipose tissue uncoupling protein 1 increased by 30%; and melanocortin 3 receptor expression declined by 60%, whereas neuropeptide Y, agouti-related protein, and MC4 receptor mRNA levels were unchanged in the NTS. In conclusion, POMC overexpression in the NTS produces a characteristic unabated hypophagia that is uniquely different from the anorexic tachyphylaxis following POMC overexpression in the hypothalamus. The sustained anorectic response may result from absence of compensatory elements in the NTS, such as increased agouti-related protein expression, suggesting melanocortin activation of the brain stem may be a viable strategy to alleviate obesity. Publication Types: Evaluation Studies Research Support, N.I.H., Extramural Research Support, U.S. Gov't, Non-P.H.S. PMID: 17389713 [PubMed - indexed for MEDLINE] 492: Anal Chim Acta. 2007 Feb 19;584(2):379-84. Epub 2006 Dec 3. Discrimination of transgenic tomatoes based on visible/near-infrared spectra. Xie L, Ying Y, Ying T, Yu H, Fu X. College of Biosystems Engineering and Food Science, Zhejiang University, 268 Kaixuan St., 310029 Hangzhou, PR China. VIS-NIR spectroscopy combined with multivariate analysis after the appropriate spectral data pre-treatment has been proved to be a very powerful tool for judgment of the relative pattern of the objects that have very similar properties. In this study, seventy transgenic tomatoes with antisense LeETR2 and 94 of their parents, non-transgenic ones were measured in VIS-NIR diffuse reflectance mode. Principal component analysis (PCA), discriminant analysis (DA) and partial least-squares discriminant analysis (PLSDA) were applied to classify tomatoes with different genes into two groups. Calibrations were developed using PLS regression with the leave-one-out cross-validation technique. The results show that differences between transgenic and non-transgenic tomatoes do exist and excellent classification can be obtained after optimizing spectral pre-treatment. The correct classifications for transgenic and non-transgenic tomatoes were both 100% using PLSDA after derivative spectral pre-treatment. The raw spectra with PLSDA model after the second derivative pre-treatment had the best satisfactory calibration and prediction abilities, with r(c)=0.97964, root mean square error of calibration (RMSEC)=0.099, r(cv)=0.97963, root mean square error of cross-validation (RMSECV)=0.0993 and a factor. The results in the present study show VIS-NIR spectroscopy together with chemometrics techniques could be used to differentiate transgenic tomato, which offers the benefit of avoiding time-consuming, costly and laborious chemical and sensory analysis. Publication Types: Research Support, Non-U.S. Gov't PMID: 17386628 [PubMed - indexed for MEDLINE] 493: Appetite. 2007 Jul;49(1):1-17. Epub 2007 Feb 24. Consumer acceptance of technology-based food innovations: lessons for the future of nutrigenomics. Ronteltap A, van Trijp JC, Renes RJ, Frewer LJ. Marketing and Consumer Behaviour Group, Wageningen University and Research Centre, Hollandseweg 1, 6706 KN Wageningen, The Netherlands. amber.ronteltap@wur.nl Determinants of consumer adoption of innovations have been studied from different angles and from the perspectives of various disciplines. In the food area, the literature is dominated by a focus on consumer concern. This paper reviews previous research into acceptance of technology-based innovation from both inside and outside the food domain, extracts key learnings from this literature and integrates them into a new conceptual framework for consumer acceptance of technology-based food innovations. The framework distinguishes 'distal' and 'proximal' determinants of acceptance. Distal factors (characteristics of the innovation, the consumer and the social system) influence consumers' intention to accept an innovation through proximal factors (perceived cost/benefit considerations, perceptions of risk and uncertainty, social norm and perceived behavioural control). The framework's application as a tool to anticipate consumer reaction to future innovations is illustrated for an actual technology-based innovation in food science, nutrigenomics (the interaction between nutrition and human genetics). Publication Types: Research Support, Non-U.S. Gov't Review PMID: 17382433 [PubMed - indexed for MEDLINE] 494: Trends Biotechnol. 2007 May;25(5):201-3. Epub 2007 Mar 19. Food from cloned animals is part of our brave old world. Miller HI. The Hoover Institution, Stanford University, Stanford, CA 94305-6010, USA. miller@hoover.stanford.edu When confronted by pressure from activists and Congress, the US Food and Drug Administration (FDA) has not always adopted policies and made decisions about individual products that accord with the scientific evidence. An example was the unnecessarily and markedly prolonged review of the veterinary drug bovine somatotropin (bST), or bovine growth hormone, during the 1980s. The FDA now faces a similar situation surrounding the question of whether meat and milk from cloned animals and their offspring are safe for human consumption. Having made a preliminary decision in the affirmative - based on an exhaustive analysis of scientific articles, health records, blood samples and studies of the composition of meat and milk - the agency has been beleaguered by criticisms. It remains to be seen whether, ultimately, science will trump anti-technology, anti-consumer activism. Publication Types: Review PMID: 17374411 [PubMed - indexed for MEDLINE] 495: FEMS Microbiol Lett. 2007 May;270(1):1-11. Epub 2007 Mar 16. Antimicrobial peptides and plant disease control. Montesinos E. Institute of Food and Agricultural Technology-CeRTA-CIDSAV, University of Girona, Girona, Spain. emonte@intea.udg.es Several diseases caused by viruses, bacteria and fungi affect plant crops, resulting in losses and decreasing the quality and safety of agricultural products. Plant disease control relies mainly on chemical pesticides that are currently subject to strong restrictions and regulatory requirements. Antimicrobial peptides are interesting compounds in plant health because there is a need for new products in plant protection that fit into the new regulations. Living organisms secrete a wide range of antimicrobial peptides produced through ribosomal (defensins and small bacteriocins) or non-ribosomal synthesis (peptaibols, cyclopeptides and pseudopeptides). Several antimicrobial peptides are the basis for the design of new synthetic analogues, have been expressed in transgenic plants to confer disease protection or are secreted by microorganisms that are active ingredients of commercial biopesticides. Publication Types: Review PMID: 17371298 [PubMed - indexed for MEDLINE] 496: J Agric Food Chem. 2007 Apr 18;55(8):2918-22. Epub 2007 Mar 20. Qualitative and quantitative polymerase chain reaction assays for an alfalfa (Medicago sativa)-specific reference gene to use in monitoring transgenic cultivars. Alexander TW, Reuter T, McAllister TA. Agriculture and Agri-Food Canada Research Centre, P.O. Box 3000, Lethbridge, Alberta, Canada. Genetically modified (GM) alfalfa (Medicago sativa) was marketed for the first time in 2005. For countries with established thresholds for GM plants, methods to detect and quantify their adventitious presence are required. We selected acetyl CoA carboxylase as a reference gene for the detection and quantification of GM alfalfa. Two qualitative polymerase chain reaction (PCR) assays (Acc1 and Acc2) were designed to detect alfalfa. Both were specific to alfalfa, amplifying DNA from 12 separate cultivars and showing negative results for PCR of 15 nonalfalfa plants. The limits of detection for Acc1 and Acc2 were 0.2 and 0.01%, respectively. A quantitative real-time PCR assay was also designed, having high linearity (r > 0.99) over alfalfa standard concentrations ranging from 100 to 2.0 x 10(5) pg of alfalfa DNA per PCR. The real-time PCR assay was effective in quantifying alfalfa DNA from forage- and concentrate-based mixed diets containing different amounts of alfalfa meal. Publication Types: Research Support, Non-U.S. Gov't PMID: 17371040 [PubMed - indexed for MEDLINE] 497: DNA Seq. 2007 Apr;18(2):131-7. Molecular cloning and characterization of ETHYLENE OVERPRODUCER 1-LIKE 1 gene, LeEOL1, from tomato (Lycopersicon esculentum Mill.) fruit. Zhu HL, Zhu BZ, Shao Y, Lin XJ, Wang XG, Gao HY, Xie YH, Li YC, Luo YB. Laboratory of Fruit Biology, College of Food Science & Nutritional Engineering, China Agricultural University, No. 17 Qinghua Donglu, Beijing 100083, People's Republic of China. Recently, ETHYLENE OVERPRODUCER 1 (ETO1) had been cloned and identified as a negative post-transcriptional regulator in the ethylene biosynthesis in Arabidopsis. However, little was known about the role of ETO1 in other species, especially in tomato, which was an ideal model for studying the biosynthesis of ethylene during tomato fruit ripening. In this study, a tomato ETHYLENE OVERPRODUCER 1-LIKE 1 (LeEOL1) was cloned. The LeEOL1 cDNA was 3,515 bp long and carried an ORF that putatively encoded a polypeptide of 886 amino acids with a predicted molecular mass of 95 kDa. It shared 74% identity in amino acid sequence with Arabidopsis EOL1 and had one BTB (Broad-complex, Tramtrack, Bric-à-brac) domain and two TPR (tetratricopeptide repeat) domains, which were also conserved domains in AtEOL1. RT-PCR analysis of the temporal expression of LeEOL1 showed that its transcript decreased companied with increase of ethylene production in tomato ripening. The level of LeEOL1 transcripts in wild type tomato fruit at mature green stage did not distinctively change when treated with exogenous ethylene. Publication Types: Research Support, Non-U.S. Gov't PMID: 17364824 [PubMed - indexed for MEDLINE] 498: Vaccine. 2007 Jul 26;25(30):5504-11. Epub 2007 Feb 26. The immunology of bovine tuberculosis and progression toward improved disease control strategies. McNair J, Welsh MD, Pollock JM. Veterinary Sciences Division, Agri-Food and Biosciences Institute, Stoney Road, Stormont, Belfast BT4 3SD, Northern Ireland, United Kingdom. jim.mcnair@afbini.gov.uk Failure to remove cattle diseased with Mycobacterium bovis has immense financial implications for disease control, animal health and agricultural trade as well as the zoonotic risk to human health. Current disease control strategies based on DTH skin testing fail to detect all diseased cattle and additional measures are urgently needed to improve detection of disease and to prevent naïve animals becoming exposed to infection. Experimental models of bovine TB traditionally based on intra-nasal instillation, intra-tracheal inoculation or placed in-contact with infected cattle, have been further developed using aerosolised bacteria delivered to the respiratory tract, allowing field-like bovine TB to be recreated under controlled, experimental conditions. Experimental infection models have already been used to improve diagnostic tests. Specificity of DTH skin testing can be improved under experimental conditions, using recombinant ESAT-6, while laboratory assays such as IFN-gamma release have benefited from the use of defined proteins to improve assay specificity. In combination, antigen cocktails may also improve test sensitivity. There is a concerted international effort to evaluate vaccines for use in cattle populations and to define vaccination strategies which will eliminate disease from infected herds. DNA, protein and genetically modified vaccines inoculated in a single dose, given as prime-boost or injected concurrently, will elicit significant protection against challenge with M. bovis under controlled conditions. However, vaccines and vaccination strategies require evaluation under field conditions. Furthermore, complementary strategies are under development to differentiate immune responses that follow vaccination from those following disease. This paper describes those recent advances which may lead to the introduction of improved disease control strategies. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 17363118 [PubMed - indexed for MEDLINE] 499: Rev Biol Trop. 2004 Sep;52(3):727-32. [Relationship of genetically modified crops with the environment and health of the Costa Rican human population] [Article in Spanish] Espinoza AM, Arrieta-Espinoza G, Sittenfeld A. Centro de Investigación en Biología Celular y Molecular, Universidad de Costa Rica, Ciudad Universitaria Rodrigo Facio, San José, Costa Rica. amespino@racsa.co.cr Genetic engineering and the food derived from genetically modified crops (GMCs) have been the center of debate worldwide, as has occurred historically with the advent of new technologies. Questions are derived from the potential impact of GMCs to the environment and the safety of the products to the consumers. In relation to the first inquiry, practice has been oriented to a case-by-case-study, according to the own characteristics of the GMC, in order to minimize its impact in the environment. Scientific studies in diverse latitudes of the world have demonstrated that GMCs in the market showed no adverse effects related to this issue. In relation to food derived from the GMCs, rigorous evaluation protocols have been developed and approved by FAO and WHO to guarantee the innocuousness of these products. Up to the moment, no contraindications for human health have been pointed out for the products that are available today in the market. In the particular case of Costa Rica, the country has established since the 90s a regulatory biosafety framework for the management of the GMCs, safeguarding the biodiversity of the country and the health of consumers. At the same time the country has made significant public and private investments in the field that allowed the country to obtain a leading position in biosafety in the region and genetic engineering research at national research centers. Any attempt to restrict or prohibit these activities in the country, will put in risk the previously described investment, will affect the generation of new knowledge for decision making and the leadership in the field, preventing the benefits derived from this promising technology. Publication Types: English Abstract PMID: 17361565 [PubMed - indexed for MEDLINE] 500: Proc Natl Acad Sci U S A. 2007 Mar 6;104(10):4218-22. Epub 2007 Mar 5. Comment in: Proc Natl Acad Sci U S A. 2007 Mar 6;104(10):3675-6. Folate biofortification of tomato fruit. Díaz de la Garza RI, Gregory JF 3rd, Hanson AD. Department of Horticultural Sciences, University of Florida, Gainesville, FL 32611, USA. Folate deficiency leads to neural tube defects and other human diseases, and is a global health problem. Because plants are major folate sources for humans, we have sought to enhance plant folate levels (biofortification). Folates are synthesized from pteridine, p-aminobenzoate (PABA), and glutamate precursors. Previously, we increased pteridine production in tomato fruit up to 140-fold by overexpressing GTP cyclohydrolase I, the first enzyme of pteridine synthesis. This strategy increased folate levels 2-fold, but engineered fruit were PABA-depleted. We report here the engineering of fruit-specific overexpression of aminodeoxychorismate synthase, which catalyzes the first step of PABA synthesis. The resulting fruit contained an average of 19-fold more PABA than controls. When transgenic PABA- and pteridine-overproduction traits were combined by crossing, vine-ripened fruit accumulated up to 25-fold more folate than controls. Folate accumulation was almost as high (up to 15-fold) in fruit harvested green and ripened by ethylene-gassing, as occurs in commerce. The accumulated folates showed normal proportions of one-carbon forms, with 5-methyltetrahydrofolate the most abundant, but were less extensively polyglutamylated than controls. Folate concentrations in developing fruit did not change in controls, but increased continuously throughout ripening in transgenic fruit. Pteridine and PABA levels in transgenic fruit were >20-fold higher than in controls, but the pathway intermediates dihydropteroate and dihydrofolate did not accumulate, pointing to a flux constraint at the dihydropteroate synthesis step. The folate levels we achieved provide the complete adult daily requirement in less than one standard serving. Publication Types: Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. PMID: 17360503 [PubMed - indexed for MEDLINE] 501: Proc Natl Acad Sci U S A. 2007 Mar 6;104(10):3675-6. Epub 2007 Mar 5. Comment on: Proc Natl Acad Sci U S A. 2007 Mar 6;104(10):4218-22. Biofortification of plant-based food: enhancing folate levels by metabolic engineering. DellaPenna D. Department of Biochemistry and Molecular Biology, Michigan State University, East Lansing, MI 48824-1319, USA. dellapen@msu.edu Publication Types: Comment PMID: 17360411 [PubMed - indexed for MEDLINE] 502: Arch Environ Contam Toxicol. 2007 May;52(4):596-602. Epub 2007 Mar 13. New analysis of a rat feeding study with a genetically modified maize reveals signs of hepatorenal toxicity. Séralini GE, Cellier D, de Vendomois JS. Committee for Independent Information and Research on Genetic Engineering CRIIGEN, Paris, France. criigen@unicaen.fr Health risk assessment of genetically modified organisms (GMOs) cultivated for food or feed is under debate throughout the world, and very little data have been published on mid- or long-term toxicological studies with mammals. One of these studies performed under the responsibility of Monsanto Company with a transgenic corn MON863 has been subjected to questions from regulatory reviewers in Europe, where it was finally approved in 2005. This necessitated a new assessment of kidney pathological findings, and the results remained controversial. An Appeal Court action in Germany (Münster) allowed public access in June 2005 to all the crude data from this 90-day rat-feeding study. We independently re-analyzed these data. Appropriate statistics were added, such as a multivariate analysis of the growth curves, and for biochemical parameters comparisons between GMO-treated rats and the controls fed with an equivalent normal diet, and separately with six reference diets with different compositions. We observed that after the consumption of MON863, rats showed slight but dose-related significant variations in growth for both sexes, resulting in 3.3% decrease in weight for males and 3.7% increase for females. Chemistry measurements reveal signs of hepatorenal toxicity, marked also by differential sensitivities in males and females. Triglycerides increased by 24-40% in females (either at week 14, dose 11% or at week 5, dose 33%, respectively); urine phosphorus and sodium excretions diminished in males by 31-35% (week 14, dose 33%) for the most important results significantly linked to the treatment in comparison to seven diets tested. Longer experiments are essential in order to indicate the real nature and extent of the possible pathology; with the present data it cannot be concluded that GM corn MON863 is a safe product. Publication Types: Research Support, Non-U.S. Gov't PMID: 17356802 [PubMed - indexed for MEDLINE] 503: Sci Am. 2007 Mar;296(3):8. The beef with cloned meat. [No authors listed] PMID: 17348147 [PubMed - indexed for MEDLINE] 504: Int J Immunopathol Pharmacol. 2007 Jan-Mar;20(1):111-8. Longer resistance of some DNA traits from BT176 maize to gastric juice from gastrointestinal affected patients. Ferrini AM, Mannoni V, Pontieri E, Pourshaban M. Istituto Superiore di Sanità, National Centre for Food Quality and Risk Assessment, Rome, Italy. ferrini@iss.it The presence of antibiotic resistance marker genes in genetically engineered plants is one of the most controversial issues related to Genetically Modified Organism (GMO)-containing food, raising concern about the possibility that these markers could increase the pool of antibiotic resistance genes. This study investigates the in vitro survival of genes bla and cryIA(b) of maize Bt176 in human gastric juice samples. Five samples of gastric juice were collected from patients affected by gastro-esophageal reflux or celiac disease and three additional samples were obtained by pH modification with NaHCO3. DNA was extracted from maize Bt176 and incubated with samples of gastric juices at different times. The survival of the target traits (bla gene, whole 1914 bp gene cry1A(b), and its 211 bp fragment) was determined using PCR. The stability of the target genes was an inverse function of their lengths in all the samples. Survival in samples from untreated subjects was below the normal physiological time of gastric digestion. On the contrary, survival time in samples from patients under anti-acid drug treatment or in samples whose pH was modified, resulted strongly increased. Our data indicate the possibility that in particular cases the survival time could be so delayed that, as a consequence, some traits of DNA could reach the intestine. In general, this aspect must be considered for vulnerable consumers (people suffering from gastrointestinal diseases related to altered digestive functionality, physiological problems or drug side-effects) in the risk analysis usually referred to healthy subjects. Publication Types: Research Support, Non-U.S. Gov't PMID: 17346434 [PubMed - indexed for MEDLINE] 505: Nat Biotechnol. 2007 Mar;25(3):282-3. Comment on: Nat Biotechnol. 2007 Jan;25(1):1. Two views of the emperor's new clones. Schubert D. Publication Types: Comment Letter PMID: 17344873 [PubMed - indexed for MEDLINE] 506: Nat Biotechnol. 2007 Mar;25(3):281. Comment on: Nat Biotechnol. 2007 Jan;25(1):1. Two views of the emperor's new clones. Miller HI. Publication Types: Comment Letter PMID: 17344872 [PubMed - indexed for MEDLINE] 507: Am J Physiol Renal Physiol. 2007 Jun;292(6):F1858-66. Epub 2007 Mar 6. Renal vascular and tubulointerstitial inflammation and proliferation in Cyp1a1-Ren2 transgenic rats with inducible ANG II-dependent malignant hypertension. Graciano ML, Mouton CR, Patterson ME, Seth DM, Mullins JJ, Mitchell KD. Department of Physiology, Hypertension and Renal Center of Excellence, Tulane University Health Sciences Center, New Orleans, Louisiana 70112, USA. Transgenic rats with inducible ANG II-dependent malignant hypertension [TGR(Cyp1a1Ren2)] were generated by inserting the mouse Ren2 renin gene into the genome of the rat. The present study was performed to assess renal morphological changes occurring during the development of ANG II-dependent malignant hypertension in these rats. Male Cyp1a1-Ren2 rats (n = 10) were fed normal rat food containing indole-3-carbinol (I3C; 0.3%) for 10 days to induce malignant hypertension. Rats induced with I3C had higher mean arterial pressures (173 +/- 9 vs. 112 +/- 11 mmHg, P < 0.01) than noninduced normotensive rats (n = 9). Glomerular damage was evaluated by determination of the glomerulosclerosis index (GSI) in tissue sections stained with periodic acid-Schiff. Kidneys of hypertensive rats had a higher GSI than normotensive rats (21.3 +/- 5.6 vs. 3.5 +/- 1.31 units). Quantitative analysis of macrophage ED-1-positive cells and proliferating cell nuclear antigen using immunohistochemistry demonstrated increased macrophage numbers in the renal interstitium (106.4 +/- 11.4 vs. 58.7 +/- 5.0 cells/mm(2)) and increased proliferating cell number in cortical tubules (37.8 +/- 5.7 vs. 24.2 +/- 2.1 cells/mm(2)), renal cortical vessels (2.2 +/- 0.5 vs. 0.13 +/- 0.07 cells/vessel), and the cortical interstitium (33.6 +/- 5.7 vs. 4.2 +/- 1.4 cells/mm(2)) of hypertensive rat kidneys. These findings demonstrate that the renal pathological changes that occur during the development of malignant hypertension in Cyp1a1-Ren2 rats are characterized by inflammation and cellular proliferation in cortical vessels and tubulointerstitium. Publication Types: Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't PMID: 17344186 [PubMed - indexed for MEDLINE] 508: Toxicol Sci. 2007 May;97(1):27-31. Epub 2007 Mar 3. The utility of an international sera bank for use in evaluating the potential human allergenicity of novel proteins. Thomas K, Bannon G, Herouet-Guicheney C, Ladics G, Lee L, Lee SI, Privalle L, Ballmer-Weber B, Vieths S. International Life Sciences Institute Health and Environmental Sciences Institute, Washington, District of Columbia 20005, USA. kthomas@ilsi.org In the safety assessment of novel foods produced through biotechnology, careful consideration is given to determining the allergenic potential of newly introduced proteins. IgE serum screening is one tool for evaluating whether the protein in question has sequence identity to a known allergen or if the source of the gene encoding the protein is a known allergenic food. A "specific" serum screen involves testing a gene product with sera from patients with documented clinical allergy to a specific allergen to confirm that the gene product of interest is not the same protein to which the patient produces IgE antibodies. A "targeted" serum screen involves testing the gene product of interest with sera from patients sensitive to food or aeroallergens from the same broad group. The concept of a global sera bank with accessible, well-characterized sera for use in such assays is an appealing option. This paper summarizes the consensus elements from a workshop to evaluate the potential utility of an international sera bank for evaluating the allergenicity of novel proteins. Areas of agreement following the workshop included the following: (1) specific sera screens are appropriate for exploring potentially cross-reactive proteins that have been identified through bioinformatics analyses; however, additional validation is needed, particularly for targeted sera screens, (2) practical and ethical considerations may preclude the formation of a global sera bank, and therefore, (3) a regional network of clinicians who could serve as sources of patient sera or be approached to conduct sera studies would be the most practical alternative. Publication Types: Congresses Research Support, Non-U.S. Gov't PMID: 17337755 [PubMed - indexed for MEDLINE] 509: J Exp Biol. 2007 Mar;210(Pt 6):956-63. The role of larval fat cells in adult Drosophila melanogaster. Aguila JR, Suszko J, Gibbs AG, Hoshizaki DK. School of Life Sciences, University of Nevada Las Vegas, 4505 Maryland Parkway, Las Vegas, NV 89154, USA. In the life history of holometabolous insects, distinct developmental stages are tightly linked to feeding and non-feeding periods. The larval stage is characterized by extensive feeding, which supports the rapid growth of the animal and allows accumulation of energy stores, primarily in the larval fat body. In Drosophila melanogaster access to these stores during pupal development is possible because the larval fat body is preserved in the pupa as individual fat cells. These larval fat cells are refractive to autophagic cell death that removes most of the larval cells during metamorphosis. The larval fat cells are thought to persist into the adult stage and thus might also have a nutritional role in the young adult. We used cell markers to demonstrate that the fat cells in the young adult are in fact dissociated larval fat body cells, and we present evidence that these cells are eventually removed in the adult by a caspase cascade that leads to cell death. By genetically manipulating the lifespan of the larval fat cells, we demonstrate that these cells are nutritionally important during the early, non-feeding stage of adulthood. We experimentally blocked cell death of larval fat cells using the GAL4/UAS system and found that in newly eclosed adults starvation resistance increased from 58 h to 72 h. Starvation survival was highly correlated with the number of remaining larval fat cells. We discuss the implications of these results in terms of the overall nutritional status of the larva as an important factor in adult survival in environmental stresses such as starvation. Publication Types: Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. PMID: 17337708 [PubMed - indexed for MEDLINE] 510: Exp Appl Acarol. 2007;41(3):191-201. Epub 2007 Mar 3. Toxicological evaluation of genetically modified cotton (Bollgard) and Dipel WP on the non-target soil mite Scheloribates praeincisus (Acari: Oribatida). Oliveira AR, Castro TR, Capalbo DM, Delalibera I Jr. Department of Entomology, Plant Pathology and Agricultural Zoology, University of Sao Paulo, CP 9, 13418-900 Piracicaba, SP, Brazil. Insecticides derived from the bacterium Bacillus thuringiensis (Bt) and plants genetically modified (GM) to express B. thuringiensis toxins are important alternatives for insect pest control worldwide. Risk assessment of B. thuringiensis toxins to non-target organisms has been extensively studied but few toxicological tests have considered soil invertebrates. Oribatid mites are one of the most diverse and abundant arthropod groups in the upper layers of soil and litter in natural and agricultural systems. These mites are exposed to the toxic compounds of GM crops or pesticides mainly when they feed on vegetal products incorporated in the soil. Although some effects of B. thuringiensis products on Acari have been reported, effects on oribatid mites are still unknown. This study investigated the effects of the ingestion of Bt cotton Bollgard and of the B. thuringiensis commercial product Dipel WP on the pantropical species Scheloribates praeincisus (Scheloribatidae). Ingestion of Bollgard and Dipel did not affect adult and immature survivorship and food consumption (estimated by number of fecal pellets produced daily) or developmental time of immature stages of S. praeincisus. These results indicate the safety of Bollgard and Dipel to S. praeincisus under field conditions where exposition is lower and other food sources besides leaves of Bt plants are available. The method for toxicological tests described here can be adapted to other species of Oribatida, consisting on a new option to risk assessment studies. Publication Types: Research Support, Non-U.S. Gov't PMID: 17334814 [PubMed - indexed for MEDLINE] 511: Food Chem Toxicol. 2007 Jul;45(7):1277-92. Epub 2007 Jan 25. Subchronic feeding study of DAS-59122-7 maize grain in Sprague-Dawley rats. Malley LA, Everds NE, Reynolds J, Mann PC, Lamb I, Rood T, Schmidt J, Layton RJ, Prochaska LM, Hinds M, Locke M, Chui CF, Claussen F, Mattsson JL, Delaney B. DuPont Haskell Laboratory, Newark, DE, United States. 59122 is a transgenic maize line containing event DAS-59122-7 that expresses the corn rootworm (CRW) specific pesticidal Cry34Ab1 and Cry35Ab1 proteins from Bacillus thuringiensis (Bt) Berliner strain PS149B1 and the phosphinothricin-N-acetyltransferase (PAT) protein from Streptomyces viridochromogenes for tolerance to the herbicidal ingredient glufosinate-ammonium. For the current study, 59122 maize grain, non-transgenic near-isogenic maize grain (091), and a commercially available non-transgenic reference maize grain (33R77) were grown under conditions simulating commercial farming practices. Adult Sprague-Dawley rats (12/sex/group) were fed diets formulated with 35% maize grain from either 59122, 091, or 33R77, or one of two separate lots of commercially available rodent chow prepared with commercially available corn (35%) in accordance with the standards of Purina Mills Labdiet 5002 for approximately 90 days. All diets possessed similar nutritional and contaminant profiles. The transgenic proteins were detected only in diets prepared with 59122 maize grain and were stable over the course of the study. Compared to control groups, no adverse diet-related differences were observed in rats fed diets formulated with 59122 maize grain with respect to body weight/gain, food consumption/efficiency, clinical signs of toxicity, mortality, ophthalmology, neurobehavioral (FOB and motor activity) assessments, clinical pathology (hematology, clinical chemistry, coagulation, and urinalysis), and pathology (organ weights and gross and microscopic pathology). Results from this study indicate that 59122 maize grain is nutritionally equivalent to and as safe as conventional maize grain. Publication Types: Research Support, Non-U.S. Gov't PMID: 17329002 [PubMed - indexed for MEDLINE] 512: Environ Biosafety Res. 2006 Apr-Jun;5(2):57-65. Epub 2007 Mar 1. A tiered system for assessing the risk of genetically modified plants to non-target organisms. Garcia-Alonso M, Jacobs E, Raybould A, Nickson TE, Sowig P, Willekens H, Van der Kouwe P, Layton R, Amijee F, Fuentes AM, Tencalla F. Syngenta, Jealott's Hill International Research Centre, Bracknell, RG42 6EY, United Kingdom. Representatives of the developers of modern agricultural biotechnology are proposing a tiered approach for conducting non-target organism risk assessment for genetically modified (GM) plants in Europe. The approach was developed by the Technical Advisory Group of the EuropaBio Plant Biotechnology Unit (http://www.europabio.org/TAG.htm) and complements other international activities to harmonize risk assessment. In the European Union (EU), the principles and methods to be followed in an environmental risk assessment for the placing on the market of GM plants are laid out in Annex II of Directive 2001/18/EC on the deliberate release into the environment of GMOs, Commission Decision 2002/623/EC and Regulation (EC) No. 1829/2003. Additional information is provided in the European Food Safety Authority guidance document of 2004. However, risk assessment for effects to non-target organisms could benefit from further clarification and remains the subject of much discussion in Europe. The industry-wide approach developed by EuropaBio is based on the fundamental steps of risk evaluation, namely hazard and exposure assessment. It follows a structured scheme including assessment planning, product characterization and assessment of hazard/exposure (Tier 0), single high dose and dose response testing (Tier 1), refined hazard characterization and exposure assessment (Tier 2) and further refined risk assessment experiments (Tier 3). An additional tier (Tier 4) was included to reflect the fact that post-market activities such as monitoring are required under Directive 2001/18/EC. The approach is compatible with conditions of commercial release in the EU and around the world. PMID: 17328852 [PubMed - indexed for MEDLINE] 513: J Agric Food Chem. 2007 Apr 4;55(7):2509-16. Epub 2007 Feb 28. Identification of PCR-amplified genetically modified organisms (GMOs) DNA by peptide nucleic acid (PNA) probes in anion-exchange chromatographic analysis. Rossi S, Lesignoli F, Germini A, Faccini A, Sforza S, Corradini R, Marchelli R. Dipartimento di Chimica Organica e Industriale Università di Parma, Viale G. P. Usberti 17/A, I-43100 Parma, Italy. PCR products obtained by selective amplification of transgenic DNA derived from food samples containing Roundup Ready soybean or Bt-176 maize have been analyzed by anion-exchange HPLC. Peptide nucleic acids (PNAs), oligonucleotide analogues known to bind to complementary single-stranded DNA with high affinity and specificity, have been used as specific probes in order to assess the identity of the peaks observed. Two different protocols were adopted in order to obtain single-stranded DNA: amplification with an excess of one primer or digestion of one DNA strand. The single-stranded DNA was mixed with the PNA probe, and the presence of a specific sequence was revealed through detection of the corresponding PNA:DNA peak with significantly different retention time. Advantages and limits of this approach are discussed. The method was tested with reference materials and subsequently applied to commercial samples. Publication Types: Research Support, Non-U.S. Gov't PMID: 17326652 [PubMed - indexed for MEDLINE] 514: Food Chem Toxicol. 2007 Jul;45(7):1179-85. Epub 2007 Jan 11. Safety assessment of transgenic Bacillus thuringiensis with VIP insecticidal protein gene by feeding studies. Peng D, Chen S, Ruan L, Li L, Yu Z, Sun M. State Key Laboratory of Agricultural Microbiology, College of Life Science and Technology, Huazhong Agricultural University, Wuhan 430070, People's Republic of China. The aim of this study was to evaluate the toxicology safety of a genetically modified (GM) Bacillus thuringiensis with vegetative insecticidal protein (VIP) gene. Acute and subacute toxicity studies by using its powder preparation were conducted in Wistar rats. The result of the acute study showed the no-observable-adverse-effect level (NOAEL) of this GM B. thuringiensis powder preparation was greater than 5000 mg/kg body weight (BW). In the subacute study, the data analysis of body weight gain, food and water consumptions, clinical observations, haematology, serum biochemistry, organ weight ratios and histopathological findings did not show significant differences between control and treated groups. These results proved the NOAEL of this GM B. thuringiensis powder preparation in subacute test was greater than 5000 mg/kg BW. Since both the acute and subacute oral toxicity were not detected at the highest dose recommended by OECD guidelines, this GM B. thuringiensis could be generally regarded as safe for use in bio-pesticide industry. Publication Types: Research Support, Non-U.S. Gov't PMID: 17320261 [PubMed - indexed for MEDLINE] 515: Vopr Pitan. 2006;75(6):55-60. [The comparative characteristic of detection methods for genetically modified organisms phytogenous] [Article in Russian] Anisimova OV, Kashina NA, Chernysheva ON, Tutel'ian VA. In the article given description, comparative characteristic and used in practice detection methods for genetically modified organisms (GMOs) in food using chip and electrophoreses. By means of these methods fined research results 704 foods that had GM analogue on the food world market. The both methods showed to identical results of research. These methods were not discovered essential distinctions in the sensibility and reliability. Publication Types: Comparative Study English Abstract PMID: 17313048 [PubMed - indexed for MEDLINE] 516: Plant Biotechnol J. 2006 Sep;4(5):529-49. Constitutive expression of the pea ABA-responsive 17 (ABR17) cDNA confers multiple stress tolerance in Arabidopsis thaliana. Srivastava S, Rahman MH, Shah S, Kav NN. Department of Agricultural, Food and Nutritional Science, University of Alberta, Edmonton, AB, Canada, T6G 2P5. The constitutive expression of the pea ABR17 (abscisic acid-responsive 17) cDNA, which is a member of the group 10 family of pathogenesis-related proteins (PR 10), in Arabidopsis thaliana is reported. The presence of ABR17 transcripts and the protein in the three transgenic lines is demonstrated by reverse transcriptase-polymerase chain reaction (RT-PCR) and two-dimensional electrophoresis followed by tandem mass spectrometry, respectively. Three independently derived transgenic lines containing ABR17 germinated better in the presence of salt, cold temperature or both. Furthermore, the transgenic plants also exhibited enhanced tolerance to freezing temperature, suggesting the potential utility of the ABR17 gene to engineer multiple stress tolerance. In order to obtain insights into the mechanism underlying ABR17-mediated stress tolerance, we have compared the proteome of a transgenic line with that of its wild-type counterpart. Several proteins were observed to be significantly altered in the transgenic line, including some with a role(s) in photosynthesis, stress tolerance and the regulation of gene expression. Our findings are discussed within the context of available genes to engineer multiple stress tolerance as well as the biological activities of the ABR17 protein. Publication Types: Research Support, Non-U.S. Gov't PMID: 17309728 [PubMed - indexed for MEDLINE] 517: Vet Rec. 2007 Feb 17;160(7):215-8. Comment in: Vet Rec. 2007 Mar 10;160(10):347; author reply 347. Bovine spongiform encephalopathy and the safety of milk from Canadian dairy cattle. Tyshenko MG. McLaughlin Centre for Population Health Risk Assessment, Institute of Population Health, University of Ottawa, 1 Stewart Street, Ottawa, Ontario, Canada K1N 6N5. The detection of bovine spongiform encephalopathy (BSE) in beef cattle closed Canadian beef export markets to 30 countries, including the USA, with devastating financial losses. The detection and confirmation of the fifth and seventh BSE-infected animals but first infected dairy cows extended the problem of risk management to Canadian dairy farmers. As the public are concerned about the safety not only of beef but also of milk and milk products that may contain disease-causing prions, this review examines the evidence for the safety of milk from studies on prions in milk or colostrum and their vertical and lateral transmission in various animal systems. The evidence indicates that the risk of contracting new variant Creutzfeldt-Jakob disease through the consumption of milk is negligible. Publication Types: Review PMID: 17308017 [PubMed - indexed for MEDLINE] 518: J Am Vet Med Assoc. 2007 Feb 15;230(4):464-6, 468. Animal clones in the food supply. Burns K. Publication Types: News PMID: 17302542 [PubMed - indexed for MEDLINE] 519: Minerva Pediatr. 2007 Feb;59(1):35-41. Rice protein-based infant formula: current status and future development. Koo WW, Lasekan JB. The Carman and Ann Adams, Department of Pediatrics, Wayne State University, Hutzel Women's Hospital, Detroit, MI 48201, USA. wkoo@wayne.edu Rice is the world's leading staple cereal food and is the major source of protein for many parts of the world. Rice is among the first solid foods fed to infants in many cultures, in part because of its hypoallergenicity from lack of gluten. Nutritional quality of rice protein compares favorably with other cereal proteins including wheat, oat and barley. It is rich in methionine and cystine, although as is the case for other cereals, it is an incomplete protein source for human infants with lysine and threonine being the primary limiting amino acids. Fortification of rice proteins with these two limiting amino acids improves its protein quality. Rice protein-based infant formulas (RPF) were initially based on high protein rice flours, but more recently are based on rice protein concentrates, isolates or hydrolysates, fortified with lysine and threonine. Hypoallergenicity efficacy, particularly for hydrolyzed rice protein-based formulas, has been reported, and limited data indicated that rice protein based infant formula may provide potentially adequate alternative if standard milk- or soy protein-based formulas are not tolerated. Unlike the rice-protein based infant formula, rice beverage formulas made from rice flour are nutritionally inadequate for infants. Reports have indicated stunted growth in infants/children fed rice beverage formulas. Future development for the RPF include those based on genetically improved rice with high lysine and threonine content, supplementation with appropriate mineral and fat blend, and long-term clinical studies in infants to confirm its efficacy and safety. PMID: 17301723 [PubMed - indexed for MEDLINE] 520: J Agric Food Chem. 2007 Feb 21;55(4):1264-73. Development of a real-time PCR method based on duplo target plasmids for determining an unexpected genetically modified soybean intermix with feed components. Dalla Costa L, Martinelli L. IASMA Research Center, Genetics and Molecular Biology Department, Via E. Mach 1, 38010 San Michele all'Adige (TN), Italy. The occurrence of intermixing, especially that resulting from genetically modified (GM) species, is increasingly becoming a problem in the delicate chain of feed and food quality control. Thus, a strategy is needed for precisely quantifying the presence of intermixing. An analytical assay based on real-time PCR has been developed; it can ascertain the extent of unexpected intermixing of GM soybean with maize meal. Three soybean-maize mix levels, with soybean intermix percentages of, respectively, 0.1, 0.5, and 1%, were prepared to simulate samples containing traces of soybean. As calibrator standards, ad hoc multiple-target pGEM-T plasmids containing soybean and maize reference genes in a 1:1 ratio were constructed. Four different maize endogenous genes, alcohol dehydrogenase 1 (adh1), high-mobility group protein a (hmga), invertase 1 (ivr1), and zein (zein), were assessed, each combined with the soybean endogenous lectin 1 (lect1) gene. Plasmids containing adh1-lect1 and zein-lect1 genes were found to be the most reliable calibration systems for this analysis, providing precise and accurate quantification results. Measuring the percentage of GM soybean intermixing makes it possible to calculate the actual transgenic component of the total sample. Publication Types: Research Support, Non-U.S. Gov't PMID: 17300150 [PubMed - indexed for MEDLINE] 521: Riv Biol. 2006 Sep-Dec;99(3):381-94. The impact of GMOs on poor countries: a threat to the achievement of the Millennium Development Goals? Francescon S. United Nations Millennium Campaign. The first of the Millennium Development Goals - halve the proportion of people who suffer from hunger by 2015 - is essential for eradicating poverty, as most of the poor live in rural areas.The role of agriculture is, therefore, key to the fight against poverty.Nevertheless, over the last years rich countries diminished their official development assistance for agricultural development and some of them proposed and pushed for a new model of agriculture based on biotechnology. Such a new model of agriculture is presented by its supporters as a means to contribute to the elimination of poverty, as it intends to maximise the crop production.However, it does not take into consideration that policies fighting hunger: need a more comprehensive approach; must take into consideration socio-economic and environmental peculiarities, especially local needs and traditional knowledge and practices.Genetically modified technology goes against these basic requirements, as it is designed to suit multinational enterprises in the North.When drafting development policies, rich and poor countries must bear in mind that the framework of the Millennium Development Goals, to which 189 Nations committed, requires a coherent approach to empower the poor, especially women, and promote traditional knowledge of indigenous people and local communities, as well as ensuring environmental sustainability.The fight to poverty and hunger will not be won and people will still go hungry if the fundamental causes of hunger and food insecurity are not tackled, whereas genetically modified technology is not based on this assumption. Publication Types: Review PMID: 17299696 [PubMed - indexed for MEDLINE] 522: J Fish Dis. 2007 Feb;30(2):65-79. Histological, digestive, metabolic, hormonal and some immune factor responses in Atlantic salmon, Salmo salar L., fed genetically modified soybeans. Bakke-McKellep AM, Koppang EO, Gunnes G, Sanden M, Hemre GI, Landsverk T, Krogdahl A. Aquaculture Protein Centre, CoE, Norway. anne.mckellep@veths.no The paper reports the second and final part of an experiment aiming to study physiological and health-related effects of genetically modified (GM) soybean meal (SBM) type Roundup Ready soybean (RRS) in diets for post-smolt Atlantic salmon. For 3 months salmon were fed diets containing 172 g kg(-1) full-fat SBM from RRS (GM-soy) or an unmodified, non-isogenic line (nGM-soy), or a reference diet with fishmeal as the sole protein source (FM). Slight differences in anti-nutrient levels were observed between the GM and nGM-soy. Histological changes were observed only in the distal intestine of the soy-fed fish. The incidence of moderate inflammation was higher in the GM-soy group (9 of 10 sampled fish) compared with the nGM-soy group (7 of 10). However, no differences in the concomitant decreases in activities of digestive enzymes located in the brush border (leucine aminopeptidase and maltase) and apical cytoplasm (acid phosphatase) of enterocytes or in the number of major histocompatibility complex class II+ cells, lysozyme activity, or total IgM of the distal intestine were observed. GM compared with nGM-soy fed fish had higher head kidney lysozyme (11,856 vs. 10,456 units g(-1) tissue) and a tendency towards higher acid phosphatase (0.45 vs. 0.39 micromol h(-1) kg(-1) body mass in whole tissue) activities, respectively. Plasma insulin and thyroxin levels, and hepatic fructose-1,6-bisphosphatase and ethoxyresorufin-O-deethylase activities were not significantly affected. It is not possible, however, to conclude whether the differences in responses to GM-soy were due to the genetic modification or to differences in soy cultivars in the soy-containing diets. Results from studies using non-modified, parental line soybeans as the control group are necessary to evaluate whether genetic modification of soybeans in diets poses any risk to farmed Atlantic salmon. Publication Types: Comparative Study Research Support, Non-U.S. Gov't PMID: 17298562 [PubMed - indexed for MEDLINE] 523: Z Naturforsch [C]. 2006 Nov-Dec;61(11-12):833-9. Effect of drought stress at supraoptimal temperature on polyamine concentrations in transgenic soybean with increased proline levels. Simon-Sarkadi L, Kocsy G, Várhegyi A, Galiba G, de Ronde JA. Department of Biochemistry and Food Technology, Budapest University of Technology and Economics, H-1521 Budapest, P.O.B. 91, Hungary. sarkadi@mail.bme.hu The effect of drought stress at supraoptimal temperature on free proline and polyamine levels was compared in wild type and transgenic soybean (Glycine max cv. Ibis) plants having increased proline levels. Since glutamate and arginine are precursors of both proline and polyamines, it was assumed that the genetic manipulation of proline levels would also affect the polyamine levels. The proline and spermine concentrations increased, while the putrescine concentration generally decreased or did not change after the treatments in both genotypes. Following drought higher proline and lower spermine levels were detected in the transgenic plants compared to the wild type ones, which could be explained by the increased use of their common precursors for proline biosynthesis in the transgenic plants. Publication Types: Research Support, Non-U.S. Gov't PMID: 17294695 [PubMed - indexed for MEDLINE] 524: Appl Microbiol Biotechnol. 2007 Apr;74(6):1175-85. Epub 2007 Feb 10. Hairy root type plant in vitro systems as sources of bioactive substances. Georgiev MI, Pavlov AI, Bley T. Institute of Food Technology and Bioprocess Engineering, Dresden University of Technology, 01069 Dresden, Germany. Milen.Georgiev@mailbox.tu-dresden.de "Hairy root" systems, obtained by transforming plant tissues with the "natural genetic engineer" Agrobacterium rhizogenes, have been known for more than three decades. To date, hairy root cultures have been obtained from more than 100 plant species, including several endangered medicinal plants, affording opportunities to produce important phytochemicals and proteins in eco-friendly conditions. Diverse strategies can be applied to improve the yields of desired metabolites and to produce recombinant proteins. Furthermore, recent advances in bioreactor design and construction allow hairy root-based technologies to be scaled up while maintaining their biosynthetic potential. This review highlights recent progress in the field and outlines future prospects for exploiting the potential utility of hairy root cultures as "chemical factories" for producing bioactive substances. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 17294182 [PubMed - indexed for MEDLINE] 525: J Allergy Clin Immunol. 2007 Apr;119(4):1013-6. Epub 2007 Feb 9. Vaccination with genetically modified birch pollen allergens: immune and clinical effects on oral allergy syndrome. Niederberger V, Reisinger J, Valent P, Krauth MT, Pauli G, van Hage M, Cromwell O, Horak F, Valenta R. Publication Types: Comparative Study Letter Multicenter Study Randomized Controlled Trial Research Support, Non-U.S. Gov't PMID: 17292956 [PubMed - indexed for MEDLINE] 526: Nat Biotechnol. 2007 Feb;25(2):169-70. Comment on: Nat Biotechnol. 2006 Jan;24(1):23-5. Model for tuning GMO detection in seed and grain. Macarthur R, Murray AW, Allnutt TR, Deppe C, Hird HJ, Kerins GM, Blackburn J, Brown J, Stones R, Hugo S. Publication Types: Comment Letter Research Support, Non-U.S. Gov't PMID: 17287745 [PubMed - indexed for MEDLINE] 527: Nat Biotechnol. 2007 Feb;25(2):165; discussion 165-6. Comment on: Nat Biotechnol. 2006 Oct;24(10):1177. Blame factory farming, not organic food. Holdrege C. Publication Types: Comment Letter PMID: 17287741 [PubMed - indexed for MEDLINE] 528: Biotechnol Prog. 2007 Mar-Apr;23(2):297-301. Epub 2007 Feb 8. Comparative evaluation of different DNA extraction procedures from food samples. Di Bernardo G, Del Gaudio S, Galderisi U, Cascino A, Cipollaro M. Department of Experimental Medicine, Section of Biotechnologies and Molecular Biology, and CRISCEB, 2nd University of Naples, Via Costantinopoli 16, 80138 Naples, Italy. Five methodologies for extracting DNA from food samples are described. The food products analyzed are from either soybean or maize. They were selected on the basis of the mechanical, thermal, and chemical treatments that they had been subjected to during industrial processing. DNA preparations were evaluated for purity, yield, and average fragment size. Two endogenous genes, soybean lectin gene and alcohol dehydrogenase gene (adh1), were used to assess the degree of DNA degradation at different stages of the transformation chain. The goal of this study was to determine the role that extraction methods play in DNA amplification in order to select the best protocol for a food sample. This comparative evaluation can be specifically useful for detection of genetically modified ingredients in a variety of food matrices. Publication Types: Comparative Study Evaluation Studies Research Support, Non-U.S. Gov't Review PMID: 17286386 [PubMed - indexed for MEDLINE] 529: Biotechnol J. 2007 Apr;2(4):486-91. Bovine fetal microchimerism in normal and embryo transfer pregnancies and its implications for biotechnology applications in cattle. Turin L, Invernizzi P, Woodcock M, Grati FR, Riva F, Tribbioli G, Laible G. Dipartimento di Patologia Animale, Igiene e Sanita' Pubblica Veterinaria, Universita' degli Studi di Milano, Milano, Italy. lauretta.turin@unimi.it Fetal cells and DNA have been detected in the maternal circulation during and after pregnancy in a few mammalian species. The incidence of similar microchimerism in cattle could have repercussion for the application of modern biotechnologies such as the transfer of transgenic embryos. To determine if feto-maternal leakage can occur in pregnant cows, we have analyzed maternal blood samples for the presence of fetal DNA during gestation and post-partum periods. Y chromosome-specific DNA was detected in up to 73% of blood samples from naturally mated heifers carrying conventional bull calves and a transgene-specific sequence in up to 50% of recipient cows carrying transgenic fetuses. These findings document for the first time that transplacental leakage of fetal DNA into the maternal circulation can occur in cattle despite the epitheliochorial placenta of ruminants, with potential implications for the utilization of recipient cows in the food chain. Publication Types: Research Support, Non-U.S. Gov't PMID: 17285678 [PubMed - indexed for MEDLINE] 530: Immunol Allergy Clin North Am. 2007 Feb;27(1):105-27. New perspectives for use of native and engineered recombinant food proteins in treatment of food allergy. Nowak-Wegrzyn A. Jaffe Food Allergy Institute, Division of Allergy and Immunology, Department of Pediatrics, Mount Sinai School of Medicine, Box 1198, One G. Levy Place, NY 10029, USA. anna.nowak-wegrzyn@mssm.edu Food allergy has emerged as an important target for research on curative treatment and prevention, with most efforts focusing on peanut, cow's milk, and egg allergy. This article reviews the recent developments in the potential treatments for IgE-mediated food allergy using native and engineered recombinant food proteins. Publication Types: Research Support, N.I.H., Extramural Review PMID: 17276882 [PubMed - indexed for MEDLINE] 531: Food Chem Toxicol. 2007 Apr;45(4):530-42. Epub 2006 Aug 25. Approaches in the risk assessment of genetically modified foods by the Hellenic Food Safety Authority. Varzakas TH, Chryssochoidis G, Argyropoulos D. Hellenic Food Safety Authority (EFET), Directorate of Nutritional Policy and Research, Karystou 5, 115 23 Ampelokipoi, Greece. theovarzakas@yahoo.gr Risk analysis has become important to assess conditions and take decisions on control procedures. In this context it is considered a prerequisite in the evaluation of GM food. Many consumers worldwide worry that food derived from genetically modified organisms (GMOs) may be unhealthy and hence regulations on GMO authorisations and labelling have become more stringent. Nowadays there is a higher demand for non-GM products and these products could be differentiated from GM products using the identity preservation system (IP) that could apply throughout the grain processing system. IP is the creation of a transparent communication system that encompasses HACCP, traceability and related systems in the supply chain. This process guarantees that certain characteristics of the lots of food (non-GM origin) are maintained "from farm to fork". This article examines the steps taken by the Hellenic Food Safety Authority to examine the presence of GMOs in foods. The whole integrated European legislation framework currently in place still needs to be implemented in Greece. Penalties should be enforced to those who import, process GMOs without special licence and do not label those products. Similar penalties should be enforced to those companies that issue false certificates beyond the liabilities taken by the food enterprises for farmers' compensation. We argue that Greece has no serious reasons to choose the use of GMOs due to the fact that the structural and pedologic characteristics of the Greek agriculture favour the biological and integrated cultivation more. Greece is not in favour of the politics behind coexistence of conventional and GM plants and objects to the use of GMOs in the food and the environment because the processor has a big burden in terms of money, time and will suffer a great deal in order to prove that their products are GMO free or that any contamination is adventitious or technically unavoidable. Moreover, Greece owns a large variety of genetic material that should try to protect from patenting and commercialisation. Finally, we should be aware of the requirements of movement of GMOs within borders, i.e. GMOs grown or used in other countries but which are not intended to cross into Greece, since Greece is very close to countries that are non-EU. This is where the development of a new, integrated, trustworthy and transparent food quality control system will help to satisfy the societal demands for safe and quality products. On the other hand, Greece should not be isolated from any recent scientific technological development and should assess the possible advantages for some cultivation using a case by case approach. Finally, the safety assessment of GM foods and feed has been discussed according to the risk assessment methodology applied by EFSA. Publication Types: Review PMID: 17275157 [PubMed - indexed for MEDLINE] 532: Am J Physiol Endocrinol Metab. 2007 May;292(5):E1483-94. Epub 2007 Jan 30. Adiposity profile in the dwarf rat: an unusually lean model of profound growth hormone deficiency. Davies JS, Gevers EF, Stevenson AE, Coschigano KT, El-Kasti MM, Bull MJ, Elford C, Evans BA, Kopchick JJ, Wells T. School of Biosciences, Cardiff University, Cardiff, UK. This study describes the previously uncharacterized ontogeny and regulation of truncal adipose reserves in the profoundly GH-deficient dwarf (dw/dw) rat. We show that, despite normal proportionate food intake, dw/dw rats develop abdominal leanness and hypoleptinemia (circulating leptin halved in dw/dw males, P < 0.05) during puberty. This contrasts with the hyperleptinemia seen in moderately GH-deficient Tgr rats (circulating leptin doubled at 6 wk of age, P < 0.05) and in GH receptor-binding protein (GHR/BP)-null mice (circulating leptin doubled; P < 0.05). This lean/hypoleptinemic phenotype was not completely normalized by GH treatment, but dw/dw rats developed abdominal obesity in response to neonatal MSG treatment or maintenance on a high-fat diet. Unlike Tgr rats, dw/dw rats did not become obese with age; plasma leptin levels and fat pad weights became similar to those in wild-type rats. In contrast with truncal leanness, tibial marrow adiposity was normal in male and doubled in female dwarves (P < 0.01), this increase being attributable to increased adipocyte number (P < 0.01). Neonatal MSG treatment and high-fat feeding elevated marrow adiposity in dw/dw rats by inducing adipocyte enlargement (P < 0.05). These results demonstrate that, despite lipolytic influence of GH, severe GH deficiency in dw/dw rats is accompanied by a paradoxical leanness. This lean/hypoleptinemic phenotype is not solely attributable to reduced GH signaling and does not appear to result from a reduction in nutrient intake or the ability of dw/dw adipocytes to accumulate lipid. Disruption of preadipocyte differentiation or adipocyte proliferation in the dw/dw rat may lead to the development of this unusually lean/hypoleptinemic phenotype. Publication Types: Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't PMID: 17264226 [PubMed - indexed for MEDLINE] 533: J Agric Food Chem. 2007 Feb 21;55(4):1274-9. Epub 2007 Jan 23. Development and evaluation of event-specific qualitative PCR methods for genetically modified Bt10 maize. Watanabe T, Tokishita S, Spiegelhalter F, Furui S, Kitta K, Hino A, Matsuda R, Akiyama H, Maitani T. National Institute of Health Sciences, 1-18-1 Kamiyoga, Setagaya-ku, Tokyo 158-8501, Japan. tawata@nihs.go.jp In 2005 it was reported that the genetically modified (GM) maize strain or "event" called Bt10 had been distributed inadvertently in the United States over the previous 4 years. In order to ensure that grain for food and feed production did not contain trace amounts of Bt10 maize and complied with the applicable regulation, highly sensitive and specific detection of Bt10 maize was required. Accordingly, we developed a novel qualitative PCR system for specific detection of Bt10 maize. Moreover, we amply evaluated the performance characteristics of two PCR systems, our own and the one provided by the developer of Bt10, Syngenta Co. Ltd. It was confirmed that both of the qualitative PCR systems can specifically detect Bt10 maize, and the results of a single-laboratory examination suggested that the limit of detection was approximately less than 0.05% for both methods. To evaluate the reproducibility of the methods, we organized an interlaboratory study with the participation of 6 laboratories and analysis of 240 blind test samples. In this paper, we report, for the first time, the statistical analysis of the qualitative PCR data obtained from the interlaboratory study. The results of this analysis also revealed that there was no significant difference in the sensitivity between the two aforementioned methods and that the limit of detection of both the methods was less than 0.05%. Thus, we conclude that both of the methods are equally suitable for correct identification and sensitive detection of the unapproved GM maize Bt10 event in test samples. Publication Types: Comparative Study Research Support, Non-U.S. Gov't PMID: 17243705 [PubMed - indexed for MEDLINE] 534: Transgenic Res. 2007 Dec;16(6):739-49. Epub 2007 Jan 20. Enhanced conversion of plant biomass into glucose using transgenic rice-produced endoglucanase for cellulosic ethanol. Oraby H, Venkatesh B, Dale B, Ahmad R, Ransom C, Oehmke J, Sticklen M. Department of Crop and Soil Sciences, Michigan State University, East Lansing, MI 48824, USA. The catalytic domain of Acidothermus cellulolyticus thermostable endoglucanase gene (encoding for endo-1,4-beta-glucanase enzyme or E1) was constitutively expressed in rice. Molecular analyses of T1 plants confirmed presence and expression of the transgene. The amount of E1 enzyme accounted for up to 4.9% of the plant total soluble proteins, and its accumulation had no apparent deleterious effects on plant growth and development. Approximately 22 and 30% of the cellulose of the Ammonia Fiber Explosion (AFEX)-pretreated rice and maize biomass respectively was converted into glucose using rice E1 heterologous enzyme. As rice is the major food crop of the world with minimal use for its straw, our results suggest a successful strategy for producing biologically active hydrolysis enzymes in rice to help generate alcohol fuel, by substituting the wasteful and polluting practice of rice straw burning with an environmentally friendly technology. Publication Types: Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. PMID: 17237981 [PubMed - indexed for MEDLINE] 535: Lab Anim. 2007 Jan;41(1):30-45. Should laboratory mice be anaesthetized for tail biopsy? Arras M, Rettich A, Seifert B, Käsermann HP, Rülicke T. Institute of Laboratory Animal Science, University of Zurich, Sternwartstrasse 6, CH-8091 Zurich, Switzerland. marras@bzl.unizh.ch Tail biopsies are routinely taken to genotype genetically modified mice. However, the effect of this procedure on the wellbeing of the animals has rarely been investigated. Thus, it has not yet been clearly demonstrated to what extent the mice suffer from tail biopsy (TB) and for how long. The aim of our study was to assess the impact of a single TB on the physiological and behavioural parameters of adult mice and to investigate whether or not anaesthesia can be beneficial. Body weight (BW) curves, daily food/water consumption and telemetric measurements of heart rate, body core temperature, and locomotor activity were recorded for three days following TB, both with and without anaesthesia with methoxyflurane (MOF) or diethylether (ether). Additionally, the impact of anaesthesia alone was characterized. TB without anaesthesia induced an increase in heart rate and locomotor activity for 1 h. Body core temperature was elevated for 2 h. In contrast, heart rate was increased for up to 4 h after anaesthesia. Body core temperature remained altered for up to 20 h after exposure to ether and for 44 h after exposure to MOF. BW was slightly reduced after MOF. Cases of death occurred exclusively under ether at a rate of 7%. Our results indicate a short-lived impact of a TB, whereas anaesthesia with either MOF or ether induced remarkable alterations in the parameters analysed. In conclusion, these types of anaesthesia did not improve mouse wellbeing following tail biopsy. PMID: 17234048 [PubMed - indexed for MEDLINE] 536: J AOAC Int. 2006 Nov-Dec;89(6):1700-1. Committee on microbiology and extraneous materials. Agin JR, Ziemer WA, Newman MC, Guilfoyle DE, Vought K, Ledenbach L, Brodsky MH, Hill W, Rice D, Ferreira JL, Werner BG, Martin BM, Shively R, Marrow T, Phillips RW, Wehling P, Labudde RA; AOAC. Committee on microbiology and extraneous materials. Ohio Department of Agriculture, 8995 E. Main St, Reynoldsburg, OH 43068, USA. Publication Types: Guideline PMID: 17225618 [PubMed - indexed for MEDLINE] 537: J Comp Physiol [B]. 2007 May;177(4):413-22. Epub 2007 Jan 16. The glutathione antioxidant system is enhanced in growth hormone transgenic coho salmon (Oncorhynchus kisutch). Leggatt RA, Brauner CJ, Iwama GK, Devlin RH. Faculty of Land and Food Systems, University of British Columbia, 2357 Main Mall, Vancouver, BC, V6T 1Z4 Canada. Insertion of a growth hormone (GH) transgene in coho salmon results in accelerated growth, and increased feeding and metabolic rates. Whether other physiological systems within the fish are adjusted to this accelerated growth has not been well explored. We examined the effects of a GH transgene and feeding level on the antioxidant glutathione and its associated enzymes in various tissues of coho salmon. When transgenic and control salmon were fed to satiation, transgenic fish had increased tissue glutathione, increased hepatic glutathione reductase activity, decreased hepatic activity of the glutathione synthesis enzyme gamma-glutamylcysteine synthetase, and increased intestinal activity of the glutathione catabolic enzyme gamma-glutamyltranspeptidase. However, these differences were mostly abolished by ration restriction and fasting, indicating that upregulation of the glutathione antioxidant system was due to accelerated growth, and not to intrinsic effects of the transgene. Increased food intake and ability to digest potential dietary glutathione, and not increased activity of glutathione synthesis enzymes, likely contributed to the higher levels of glutathione in transgenic fish. Components of the glutathione antioxidant system are likely upregulated to combat potentially higher reactive oxygen species production from increased metabolic rates in GH transgenic salmon. Publication Types: Research Support, Non-U.S. Gov't PMID: 17225138 [PubMed - indexed for MEDLINE] 538: Nature. 2007 Jan 11;445(7124):132-3. Out of bounds. [No authors listed] Publication Types: News PMID: 17215811 [PubMed - indexed for MEDLINE] 539: J Mol Med. 2007 Apr;85(4):405-13. Epub 2007 Jan 9. Erratum in: J Mol Med. 2007 Apr;85(4):421. Copper and clioquinol treatment in young APP transgenic and wild-type mice: effects on life expectancy, body weight, and metal-ion levels. Schäfer S, Pajonk FG, Multhaup G, Bayer TA. Department of Psychiatry, Division of Neurobiology, Saarland University, Homburg, Germany. There is mounting evidence that the amyloid precursor protein (APP), the key protein in Alzheimer's disease (AD) is involved in the copper (Cu) homeostasis in the brain. Conflicting results about the potential use of dietary Cu and clioquinol (CQ), a known Cu chelator, have been reported using APP transgenic mice. Previously, in vitro studies have demonstrated that CQ can act as a Cu transporter. To analyze the potential function of CQ as a Cu transporter in vivo, the nutritional effect of Cu and CQ was analyzed in young APP transgenic mice and nontransgenics with food pellets containing either Cu, CQ, Cu plus CQ (Cu + CQ), or without addition of supplements (control). The offspring were fed with corresponding food pellets until the age of 14 weeks. We observed an increased lethality of APP transgenics upon CQ treatment, which could be rescued by a co-treatment with Cu. The exposure of Cu + CQ led to a modest but significant increase in cerebral Cu levels, most likely due to an enhanced transport of CQ-Cu complexes. In CQ or Cu + CQ treatment groups, the plasma levels of Cu, zinc, and iron were reduced in all animals; moreover, Cu treatment alone reduced only plasma iron levels. We conclude not only that CQ has certain toxicity but also that the chelating effect, perhaps, plays a secondary role with respect to its properties as an intracellular Cu transporter, thus, counteracting the supposed therapeutic effects of CQ as an agent for chelating therapy in AD. Publication Types: Research Support, Non-U.S. Gov't PMID: 17211610 [PubMed - indexed for MEDLINE] 540: Nat Biotechnol. 2007 Jan;25(1):77-83. Risk assessment of meat and milk from cloned animals. Yang X, Tian XC, Kubota C, Page R, Xu J, Cibelli J, Seidel G Jr. Center for Regenerative Biology and Department of Animal Science, University of Connecticut, Storrs, Connecticut 06269-4243, USA. xiangzhong.yang@uconn.edu Research on, and commercialization of, cloned cattle has been conducted for more than 20 years. Early techniques relied on the physical splitting of embryos or using embryo cells for nuclear transfer to generate cloned animals. Milk and meat from these animals entered into the human food market with no evidence of problems. With the advent of nuclear transfer, which enables the direct transference and preservation of high-value meat- and milk-producing genotypes to offspring, concerns have been raised about whether the products from somatic cell nuclear transfer-produced animals are safe for human consumption. Studies on the biochemical properties of food products from cloned and noncloned animals have thus far not detected any differences. All data to date indicate no significant differences in the measured parameters between animals created by nuclear transfer and normally bred animals. Public acceptance of cloned animal products depends upon forthcoming US Food and Drug Administration approval along with convincing safety data. Publication Types: Review PMID: 17211406 [PubMed - indexed for MEDLINE] 541: Nat Biotechnol. 2007 Jan;25(1):47-53. Dolly for dinner? Assessing commercial and regulatory trends in cloned livestock. Suk J, Bruce A, Gertz R, Warkup C, Whitelaw CB, Braun A, Oram C, Rodríguez-Cerezo E, Papatryfon I. ESRC Genomics Policy & Research Forum, University of Edinburgh, St. John's Land, Edinburgh, Scotland. As cloning technologies become more widely established, will products enter the food chain sooner than regulatory agencies and the public might be prepared for? Publication Types: Research Support, Non-U.S. Gov't PMID: 17211395 [PubMed - indexed for MEDLINE] 542: Nat Biotechnol. 2007 Jan;25(1):39-43. Animal cloning and the FDA--the risk assessment paradigm under public scrutiny. Rudenko L, Matheson JC, Sundlof SF. Center for Veterinary Medicine, US Food and Drug Administration, Department of Health and Human Services, 7500 Standish Place, Rockville, Maryland 20855, USA. larisa.rudenko@fda.hhs.gov The evidence gathered thus far--ultimately to be published in the Draft Risk Assessment on Animal Cloning--indicates that there are no unique risks associated with animal cloning. Publication Types: Review PMID: 17211392 [PubMed - indexed for MEDLINE] 543: Nat Biotechnol. 2007 Jan;25(1):35-6; author reply 36-7. Comment on: Nat Biotechnol. 2005 Dec;23(12):1475-6. Nat Biotechnol. 2006 Jan;24(1):63-71. Early-tier tests insufficient for GMO risk assessment. Lang A, Lauber E, Darvas B. Publication Types: Comment Letter PMID: 17211390 [PubMed - indexed for MEDLINE] 544: Nat Biotechnol. 2007 Jan;25(1):33-4. Comment on: Nat Biotechnol. 2006 Oct;24(10):1178. Parallel biopolitical universes. Morris SH. Publication Types: Comment Letter PMID: 17211389 [PubMed - indexed for MEDLINE] 545: Nat Biotechnol. 2007 Jan;25(1):7-8. Epub 2007 Jan 5. FDA's cloning report bypasses ethics, exposes European dilemma. Vermij P. Publication Types: News PMID: 17211377 [PubMed - indexed for MEDLINE] 546: Nat Biotechnol. 2007 Jan;25(1):1. Comment in: Nat Biotechnol. 2007 Mar;25(3):281. Nat Biotechnol. 2007 Mar;25(3):282-3. The emperor's new clones. [No authors listed] If regulators conclude that food from clones poses no more risk than food from other animals, the US and Europe could be on course for another biotech trade war. Publication Types: Editorial PMID: 17211372 [PubMed - indexed for MEDLINE] 547: Plant Biotechnol J. 2007 Jan;5(1):109-17. Doubled sugar content in sugarcane plants modified to produce a sucrose isomer. Wu L, Birch RG. Botany Department - SIB, The University of Queensland, Brisbane, Qld 4072, Australia. Sucrose is the feedstock for more than half of the world's fuel ethanol production and a major human food. It is harvested primarily from sugarcane and beet. Despite attempts through conventional and molecular breeding, the stored sugar concentration in elite sugarcane cultivars has not been increased for several decades. Recently, genes have been cloned for bacterial isomerase enzymes that convert sucrose into sugars which are not metabolized by plants, but which are digested by humans, with health benefits over sucrose. We hypothesized that an appropriate sucrose isomerase (SI) expression pattern might simultaneously provide a valuable source of beneficial sugars and overcome the sugar yield ceiling in plants. The introduction of an SI gene tailored for vacuolar compartmentation resulted in sugarcane lines with remarkable increases in total stored sugar levels. The high-value sugar isomaltulose was accumulated in storage tissues without any decrease in stored sucrose concentration, resulting in up to doubled total sugar concentrations in harvested juice. The lines with enhanced sugar accumulation also showed increased photosynthesis, sucrose transport and sink strength. This remarkable step above the former ceiling in stored sugar concentration provides a new perspective into plant source-sink relationships, and has substantial potential for enhanced food and biofuel production. Publication Types: Research Support, Non-U.S. Gov't PMID: 17207261 [PubMed - indexed for MEDLINE] 548: Plant Biotechnol J. 2007 Jan;5(1):50-9. Therapeutic effectiveness of orally administered transgenic low-alkaloid tobacco expressing human interleukin-10 in a mouse model of colitis. Menassa R, Du C, Yin ZQ, Ma S, Poussier P, Brandle J, Jevnikar AM. Southern Crop Protection and Food Research Centre, Agriculture and Agri-Food Canada, 1391 Sandford St., London, ON, Canada, N5V 4T3. Inflammatory bowel disease (IBD) represents a spectrum of diseases in which inflammation leads to acute and chronic gut injury. It is a growing health issue for which no cure exists. The pathogenesis is multifactorial with links to infectious and environmental events that trigger disease in genetically predisposed individuals. Treatment of the two major forms of IBD, Crohn's disease and ulcerative colitis, involves the reduction of inflammation with toxic immunosuppressive drugs or blocking of the pro-inflammatory effects of tumour necrosis factor-alpha (TNF-alpha) with antibodies. Here, we show that the oral administration of transgenic low-alkaloid tobacco expressing the contra-inflammatory cytokine human interleukin-10 (hIL-10) reduces the severity of colitis by down-regulating TNF-alpha expression directly at the sites of inflammation in IBD-susceptible IL-10(-/-) mice. hIL-10 expressed in plants is biologically active and displays resistance to gastrointestinal degradation. Dietary supplementation with plant tissue delivering up to 9 microg of hIL-10 daily for 4 weeks was well tolerated by treated mice. Gut histology was significantly improved relative to controls (P = 0.002), and was correlated with a decrease in small bowel TNF-alpha mRNA levels and an increase in IL-2 and IL-1beta mRNA levels. Transgenic plants expressing IL-10 to directly attenuate TNF-alpha expression at sites of inflammation in the gut may become a useful new approach in the luminal therapy of IBD. Publication Types: Research Support, Non-U.S. Gov't PMID: 17207256 [PubMed - indexed for MEDLINE] 549: Ig Sanita Pubbl. 2005 Sep-Oct;61(5):475-96. [Genetically modified organisms: European and Italian legislation to protect citizens' health] [Article in Italian] Sotgiu A, Tala M, Sardu G, Coroneo V, Dessi S, Contu P. Dipartimento di Sanità Pubblica, Università di Cagliari. The development of GM foods and organisms has concentrated everyone's attention on the importance of food safety and on protecting citizens' health, and inevitably influenced healthcare policies regarding food safety. Personal ethical beliefs regarding food and in particular, the consumption of foods derived from biotechnology should be taken into account when deciding healthcare policy. AIM: The aim of this study was to analyse whether European, Italian and Regional legislation meets basic human rights regarding health and the right to choose, based on the precautionary principle. METHODS: European and Italian laws regarding the production and marketing of GM foods were analysed and compared to food safety legislation, in order to evaluate how and to what degree existing legislation protects consumers' right to choose. Results show that existing legislation protects consumers from possible foodborne diseases, but the right to informed consent and to free choice is not warranted. Existing laws do not attach enough importance to consumers' right to information; arbitrary threshold levels set for labeling and clauses concerning technical causes allow food businesses to avoid labeling and do not give consumers the possibility of making an informed choice. Publication Types: English Abstract PMID: 17206218 [PubMed - indexed for MEDLINE] 550: EMBO J. 2007 Jan 24;26(2):448-58. Epub 2007 Jan 4. Nutrient starvation promotes condensin loading to maintain rDNA stability. Tsang CK, Li H, Zheng XS. Department of Pharmacology, Robert Wood Johnson Medical School, Piscataway, NJ 08854, USA. Nutrient starvation or rapamycin treatment, through inhibition of target of rapamycin, causes condensation of ribosomal DNA (rDNA) array and nucleolar contraction in budding yeast. Here we report that under such conditions, condensin is rapidly relocated into the nucleolus and loaded to rDNA tandem repeats, which is required for rDNA condensation. Rpd3-dependent histone deacetylation is necessary and sufficient for condensin's relocalization and loading to rDNA array, suggesting that histone modification plays a regulatory role for condensin targeting. Rapamycin independently, yet coordinately, inhibits rDNA transcription and promotes condensin loading to rDNA array. Unexpectedly, we found that inhibition of rDNA transcription in the absence of condensin loading leads to rDNA instability. Our data suggest that enrichment of condensin prevents rDNA instability during nutrient starvation. Together, these observations unravel a novel role for condensin in the maintenance of regional genomic stability. Publication Types: Research Support, N.I.H., Extramural PMID: 17203076 [PubMed - indexed for MEDLINE] 551: Rapid Commun Mass Spectrom. 2007;21(3):319-28. Mass spectrometric detection of CP4 EPSPS in genetically modified soya and maize. Ocaña MF, Fraser PD, Patel RK, Halket JM, Bramley PM. Centre for Chemical and Bioanalytical Sciences, Royal Holloway, University of London, Egham TW20 0EX, UK. The potential of protein fractionation hyphenated to mass spectrometry (MS) to detect and characterize the transgenic protein present in Roundup Ready soya and maize has been investigated. Genetically modified (GM) soya and maize contain the 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) gene from Agrobacterium tumefaciens CP4, which confers resistance to the herbicide glyphosate. The GM soya and maize proteomes were fractionated by gel filtration, anion-exchange chromatography and sodium dodecyl sulfate/polyacrylamide gel electrophoresis (SDS-PAGE) prior to MS. This facilitated detection of a tryptic peptide map of CP4 EPSPS by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) MS and nanoelectrospray ionization quadrupole time-of-flight (nanoESI-QTOF) MS. Subsequently, sequence information from the CP4 EPSPS tryptic peptides was obtained by nanoESI-QTOF MS/MS. The identification was accomplished in 0.9% GM soya seeds, which is the current EU threshold for food-labeling requirements. Copyright 2007 John Wiley & Sons, Ltd. Publication Types: Research Support, Non-U.S. Gov't PMID: 17200978 [PubMed - indexed for MEDLINE] 552: Recent Pat Biotechnol. 2007;1(1):75-97. Nutraceuticals, nutritional therapy, phytonutrients, and phytotherapy for improvement of human health: a perspective on plant biotechnology application. Zhao J. Children's Nutrition Research Center, Department of Pediatrics, Baylor College of Medicine, Houston, TX 77030, USA. jzhao1@bcm.tmc.edu Plants are one of the most important resources of human foods and medicines. Rapidly increasing knowledge on nutrition, medicine, and plant biotechnology has dramatically changed the concepts about food, health and agriculture, and brought in a revolution on them. Nutritional therapy and phytotherapy have emerged as new concepts and healing systems have quickly and widely spread in recent years. Strong recommendations for consumption of nutraceuticals, natural plant foods, and the use of nutritional therapy and phytotherapy have become progressively popular to improve health, and to prevent and treat diseases. With these trends, improving the dietary nutritional values of fruits, vegetables and other crops or even bioactive components in folk herbals has become targets of the blooming plant biotechnology industry. This review attempts to display and remark on these aspects. It summarizes the progress made on nutraceuticals, nutritional therapy, phytonutrients, phytotherapy, and their related epidemiological investigations and clinical studies. It also covers markets of these health-promoting products and disease-preventing or healing systems, as well as regulations behind them that direct the development of biotechnology study and application. Finally, related patents are listed and briefly analyzed, regarding of plant biotechnological research and progress on transgenic crops to improve nutritional value, phytotherapy efficiency, or to produce pharmaceutically important secondary metabolites or high-valued protein medicines such as vaccines and antibodies. Publication Types: Review PMID: 19075834 [PubMed - indexed for MEDLINE] 553: Biotechniques. 2006 Dec;41(6):708-10. Minimal T-DNA vectors suitable for agricultural deployment of transgenic plants. Barrell PJ, Conner AJ. National Centre for Advanced Bio-Protection Technologies and New Zealand Institute for Crop & Food Research Ltd, Canterbury, New Zealand. barrellp@crop.cri.nz PMID: 17191614 [PubMed - indexed for MEDLINE] 554: J Biosci Bioeng. 2006 Nov;102(5):375-89. Developments in biotechnological production of sweet proteins. Masuda T, Kitabatake N. Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Uji, Kyoto 611-0011, Japan. Most proteins are tasteless and flavorless, while some proteins elicit a sweet-taste response on the human palate. Six proteins, thaumatin, monellin, mabinlin, brazzein, egg lysozyme, and neoculin (previously considered as curculin) have been identified as sweet-tasting proteins. However, no common features among them have been observed. Herein, recent advances in the research of sweet-tasting proteins and the production of such proteins by biotechnological approaches are reviewed. Information on the structure-sweetness relationship for these proteins would help not only in the clarification of the mechanism of interaction of sweet-tasting proteins with their receptors, but also in the design of more effective low-calorie sweeteners. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 17189164 [PubMed - indexed for MEDLINE] 555: Risk Anal. 2006 Dec;26(6):1707-19. Exploring the structure of attitudes toward genetically modified food. Poortinga W, Pidgeon NF. Cardiff University, Welsh School of Architecture, Cardiff, Wales, UK. PoortingaW@Cardiff.ac.uk Although it is often thought that the British public is opposed to genetically modified (GM) food, recent qualitative work suggests that most people are ambivalent about GM food and crops. In this article we explore the structure of attitudes in order to examine whether attitudinal ambivalence can be captured by more quantitative methods. Based on the finding that the perceived risks and benefits of GM food can be treated as independent dimensions, we propose a four-way typology of attitudes, consisting of a positive, negative, indifferent, and ambivalent group. This study showed that the differences between the four groups could best be described by three main dimensions: (1) a general evaluative dimension, (2) an involvement dimension, and (3) an attitudinal certainty dimension. While these different attitudinal dimensions have generally been studied in isolation, we argue that they should be studied collectively. Publication Types: Research Support, Non-U.S. Gov't PMID: 17184407 [PubMed - indexed for MEDLINE] 556: Br J Nutr. 2006 Dec;96(6):997-1005. Conventional and real-time polymerase chain reaction assessment of the fate of transgenic DNA in sheep fed Roundup Ready rapeseed meal. Alexander TW, Reuter T, Okine E, Sharma R, McAllister TA. Agriculture and Agri-Food Canada Research Centre, Lethbridge, Alberta, Canada. Conventional and real-time PCR were used to detect transgenic DNA in digesta, faeces and blood collected from six ruminally and duodenally cannulated sheep fed forage-based (F) or concentrate-based (C) diets containing 15% Roundup Ready (RR) rapeseed meal (n 3). The sheep were adapted for 14 d to F or C diets containing non-GM rapeseed, then fed the RR diets for 11 d. On day 12, they were switched back to non-GM diets for a further 11 d. Ruminal and duodenal fluids (RF, DF) and faecal samples were collected at 3 or 4 h intervals over the 4 d immediately following the last feeding of GM diets. DNA was isolated from whole RF and DF, from the cell-free supernatant fraction, and from culture fermentation liquid. Blood was collected on days 1, 5 and 9 of feeding the RR rapeseed meal. The 1363 bp 5-enolpyruvylshikimate-3-phosphate synthase transgene (epsps) was quantifiable in whole RF and DF for up to 13 h, and a 108 bp epsps fragment for up to 29 h. Transgenic DNA was not detectable in faeces or blood, or in microbial DNA. Diet type (F v. C) did not affect (P>0.05) the quantity of transgenic DNA in digesta. More (P<0.05) transgenic DNA was detected in RF than in DF, but there was an interaction (P<0.05) between sample type and collection time. In supernatant fractions from RF and DF, three different fragments of transgenic DNA ranging in size from 62 to 420 bp were not amplifiable. PMID: 17181873 [PubMed - indexed for MEDLINE] 557: Plant Biotechnol J. 2006 Jul;4(4):433-44. Pathway engineering for healthy phytochemicals leading to the production of novel flavonoids in tomato fruit. Schijlen E, Ric de Vos CH, Jonker H, van den Broeck H, Molthoff J, van Tunen A, Martens S, Bovy A. Plant Research International, Business Unit Bioscience, PO Box 16, 6700 AA Wageningen, The Netherlands. elio.schijlen@wur.nl Flavonoids are a large family of plant polyphenolic secondary metabolites. Although they are widespread throughout the plant kingdom, some flavonoid classes are specific for only a few plant species. Due to their presumed health benefits there is growing interest in the development of food crops with tailor-made levels and composition of flavonoids, designed to exert an optimal biological effect. In order to explore the possibilities of flavonoid engineering in tomato fruits, we have targeted this pathway towards classes of potentially healthy flavonoids which are novel for tomato. Using structural flavonoid genes (encoding stilbene synthase, chalcone synthase, chalcone reductase, chalcone isomerase and flavone synthase) from different plant sources, we were able to produce transgenic tomatoes accumulating new phytochemicals. Biochemical analysis showed that the fruit peel contained high levels of stilbenes (resveratrol and piceid), deoxychalcones (butein and isoliquiritigenin), flavones (luteolin-7-glucoside and luteolin aglycon) and flavonols (quercetin glycosides and kaempferol glycosides). Using an online high-performance liquid chromatography (HPLC) antioxidant detection system, we demonstrated that, due to the presence of the novel flavonoids, the transgenic tomato fruits displayed altered antioxidant profiles. In addition, total antioxidant capacity of tomato fruit peel with high levels of flavones and flavonols increased more than threefold. These results on genetic engineering of flavonoids in tomato fruit demonstrate the possibilities to change the levels and composition of health-related polyphenols in a crop plant and provide more insight in the genetic and biochemical regulation of the flavonoid pathway within this worldwide important vegetable. Publication Types: Research Support, Non-U.S. Gov't PMID: 17177808 [PubMed - indexed for MEDLINE] 558: Plant Biotechnol J. 2006 Mar;4(2):263-73. Characterization of a higher plant herbicide-resistant phytoene desaturase and its use as a selectable marker. Arias RS, Dayan FE, Michel A, Howell J, Scheffler BE. USDA-ARS, Natural Products Utilization Research Unit, PO Box 8048, University, MS 38677, USA. Three natural somatic mutations at codon 304 of the phytoene desaturase gene (pds) of Hydrilla verticillata (L. f. Royle) have been reported to provide resistance to the herbicide fluridone. We substituted the arginine 304 present in the wild-type H. verticillata phytoene desaturase (PDS) with all 19 other natural amino acids and tested PDS against fluridone. In in vitro assays, the threonine (Thr), cysteine (Cys), alanine (Ala) and glutamine (Gln) mutations imparted the highest resistance to fluridone. Thr, the three natural mutations [Cys, serine (Ser), histidine (His)] and the wild-type PDS protein were tested in vitro against seven inhibitors of PDS representing several classes of herbicides. These mutations conferred cross-resistance to norflurazon and overall negative cross-resistance to beflubutamid, picolinafen and diflufenican. The T3 generation of transgenic Arabidopsis thaliana plants harbouring the four selected mutations and wild-type pds had similar patterns of cross-resistance to the herbicides as observed in the in vitro assays. The Thr304 Hydrilla pds mutant proved to be an excellent marker for the selection of transgenic plants. Seedlings harbouring Thr304 pds had a maximum resistance to sensitivity (R/S) ratio of 57 and 14 times higher than that of the wild-type for treatments with norflurazon and fluridone, respectively. These plants exhibited normal growth and development, even after long-term exposure to herbicide. As Thr304 pds is of plant origin, it could become more acceptable than other selectable markers for use in genetically modified food. PMID: 17177802 [PubMed - indexed for MEDLINE] 559: Plant Biotechnol J. 2006 Jan;4(1):123-34. Manipulation of starch granule size distribution in potato tubers by modulation of plastid division. de Pater S, Caspers M, Kottenhagen M, Meima H, ter Stege R, de Vetten N. TNO Nutrition and Food Research, Department of Applied Plant Sciences, Zernikedreef 9, 2333 CK Leiden, the Netherlands. pater@rulbim.leidenuniv.nl Starch granule size is an important parameter for starch applications in industry. Starch granules are formed in amyloplasts, which are, like chloroplasts, derived from proplastids. Division processes and associated machinery are likely to be similar for all plastids. Essential roles for FtsZ proteins in plastid division in land plants have been revealed. FtsZ forms the so-called Z ring which, together with inner and outer plastid division rings, brings about constriction of the plastid. It has been shown that modulation of the expression level of FtsZ may result in altered chloroplast size and number. To test whether FtsZ is also involved in amyloplast division and whether this, in turn, may affect the starch granule size in crop plants, FtsZ protein levels were either reduced or increased in potato. As shown previously in other plant species, decreased StFtsZ1 protein levels in leaves resulted in a decrease in the number of chloroplasts in guard cells. More interestingly, plants with increased StFtsZ1 protein levels in tubers resulted in less, but larger, starch granules. This suggests that the stoichiometry between StFtsZ1 and other components of the plastid division machinery is important for its function. Starch from these tubers also had altered pasting properties and phosphate content. The importance of our results for the starch industry is discussed. Publication Types: Research Support, Non-U.S. Gov't PMID: 17177791 [PubMed - indexed for MEDLINE] 560: J Agric Food Chem. 2006 Dec 27;54(26):9901-5. Biochemical safety evaluation of transgenic rice seeds expressing T cell epitopes of Japanese cedar pollen allergens. Takagi H, Hirose S, Yasuda H, Takaiwa F. Transgenic Crop Research and Development Center, National Institute of Agrobiological Sciences, 2-1-2 Kannondai, Tsukuba, Ibaraki 305-8602, Japan. Transgenic rice seeds, which express a hybrid peptide comprising seven predominant human T cell epitopes (7Crp) derived from Japanese cedar pollen allergens, have been shown to function as an effective edible vaccine for the control of pollen allergen-induced responses. In this study, we characterized biochemical properties of transgenic seeds expressing the 7Crp peptide. The levels of chemical compositions, such as carbohydrate, protein, lipid, amino acid, fatty acid, mineral, and vitamin, were substantially equivalent between transgenic 7Crp and its nontransgenic counterpart seeds. The contents of three major allergenic proteins in transgenic seeds were not enhanced by expression of the 7Crp peptide when compared with those of nontransgenic seeds. The 7Crp peptide expressed in seeds was susceptible to simulated gastric/intestinal fluids. N-Glycosylation was not observed in the 7Crp peptide sequence. These results indicate that transgenic 7Crp seeds are substantially equivalent to nontransgenic parental seeds except for the presence of the 7Crp peptide. Keywords: Food safety assessment; transgenic rice seed; edible vaccine; peptide-based immunotherapy; Japanese cedar pollinosis. Publication Types: Research Support, Non-U.S. Gov't PMID: 17177518 [PubMed - indexed for MEDLINE] 561: J Agric Food Chem. 2006 Dec 27;54(26):9882-7. Improving potato storage and processing characteristics through all-native DNA transformation. Rommens CM, Ye J, Richael C, Swords K. J. R. Simplot Company, Simplot Plant Sciences, Boise, Idaho 83706, USA. The dominant potato (Solanum tuberosum) variety for French fry production in the United States is the 131-year-old Russet Burbank. Market penetration of the higher yielding and more uniform Ranger Russet variety is limited to about one-fifth of that of the Russet Burbank because of two storage deficits: black spot bruise sensitivity and high levels of cold-induced sweetening. Here, these trait weaknesses are turned into strengths by simultaneously lowering the expression of Ranger Russet's tuber-expressed polyphenol oxidase (Ppo), starch-associated R1, and phosphorylase-L (PhL) genes. This genetic modification was accomplished without inserting any foreign DNA into the plant genome. French fries from the intragenic potatoes also contained reduced amounts of the antinutritional compound acrylamide while, unexpectedly, displaying enhanced sensory characteristics. PMID: 17177515 [PubMed - indexed for MEDLINE] 562: J Agric Food Chem. 2006 Dec 27;54(26):9658-63. Equal performance of TaqMan, MGB, molecular beacon, and SYBR green-based detection assays in detection and quantification of roundup ready soybean. Andersen CB, Holst-Jensen A, Berdal KG, Thorstensen T, Tengs T. Section of Food and Feed Microbiology, National Veterinary Institute, Ullevaalsveien 68, 0454 Oslo, Norway. We have tested and compared the performance of 12 different assays representing four different real-time polymerase chain reaction (PCR) chemistries in the context of genetically modified organism detection. Several different molecular beacon, SYBR Green, TaqMan, and MGB assays were designed for the event specific detection and quantification of the 3' integration junction of GTS 40-3-2 (Roundup Ready) soybean. Sensitivity as well as robustness in the presence of background DNA were tested. None of the PCR-based approaches appeared to be significantly better than any of the other, but the molecular beacon assays had the lowest efficiency and also seemed more sensitive to changes in experimental setup. Publication Types: Comparative Study Research Support, Non-U.S. Gov't PMID: 17177484 [PubMed - indexed for MEDLINE] 563: Adv Exp Med Biol. 2007;591:1-13. Nuclear remodeling and nuclear reprogramming for making transgenic pigs by nuclear transfer. Prather RS. Division of Animal Science, Food for the 21st Century, College of Food, Agriculture & Natural Resources, University of Missouri-Columbia, 920 East Campus Drive, E125 ASRC, Columbia, Missouri 65211-5300, USA. PratherR@missouri.edu A better understanding of the cellular and molecular events that occur when a nucleus is transferred to the cytoplasm of an oocyte will permit the development of improved procedures for performing nuclear transfer and cloning. In some cases it appears that the gene(s) are reprogrammed, while in other cases there appears to be little effect on gene expression. Not only does the pattern of gene expression need to be reprogrammed, but other structures within the nucleus also need to be remodeled. While nuclear transfer works and transgenic and knockout animals can be created, it still is an inefficient process. However, even with the current low efficiencies this technique has proved very valuable for the production of animals that might be useful for tissue or organ transplantation to humans. Publication Types: Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Review PMID: 17176551 [PubMed - indexed for MEDLINE] 564: Insect Biochem Mol Biol. 2007 Jan;37(1):10-8. Epub 2006 Oct 4. The cyanogenic glucoside composition of Zygaena filipendulae (Lepidoptera: Zygaenidae) as effected by feeding on wild-type and transgenic lotus populations with variable cyanogenic glucoside profiles. Zagrobelny M, Bak S, Ekstrøm CT, Olsen CE, Møller BL. Department of Plant Biology and Center for Molecular Plant Physiology (PlaCe), 40 Thorvaldsensvej, DK-1871 Frederiksberg C, Copenhagen, Denmark. miz@kvl.dk Zygaena larvae sequester the cyanogenic glucosides linamarin and lotaustralin from their food plants (Fabaceae) as well as carry out de novo biosynthesis of these compounds. In this study, Zygaena filipendulae were reared on wild-type Lotus corniculatus and wild-type and transgenic L. japonicus plants with differing content and ratios of the cyanogenic glucosides linamarin and lotaustralin and of the cyanoalkenyl glucosides rhodiocyanoside A and D. LC-MS analyses, free choice feeding experiments and developmental studies were used to examine the effect of varying content and ratios of these secondary metabolites on the feeding preferences, growth and development of Z. filipendulae. Larvae reared on cyanogenic L. corniculatus developed faster compared to larvae reared on L. japonicus although free choice feeding trials demonstrated that the latter plant source was the preferred food plant. Larvae reared on acyanogenic L. corniculatus showed decelerated development. Analysis of different life stages and tissues demonstrate that Z. filipendulae strive to maintain certain threshold content and ratios of cyanogenic glucosides regardless of the composition of the food plants. Despite this, the ratios of cyanogenic glucosides in Z. filipendulae remain partly affected by the ratio of the food plant due to the high proportion of sequestering that takes place. PMID: 17175442 [PubMed - indexed for MEDLINE] 565: Methods Mol Biol. 2007;354:183-95. Methods for engineering resistance to plant viruses. Sudarshana MR, Roy G, Falk BW. Western Institute for Food Safety and Security, University of California, Davis, USA. The development of genetically engineered resistance to plant viruses is a result of efforts to understand the plant-virus interactions involved in "crossprotection," a phenomenon observed with several plant virus diseases. Historically, expression of the coat protein gene of Tobacco mosaic virus in transgenic tobacco (Nicotiana tabacum) plants is the first example of transgene-mediated resistance to a plant virus. Subsequently, virus-derived sequences of several plant viruses were shown to confer virus resistance in experimental and/or natural hosts. For plant RNA viruses, virus complementary DNA sequences shown to confer resistance include wild-type genes, mutated genes that produced truncated protein products, and nontranslatable sense or antisense transcripts to various regions of the virus genome. Resistance also has been demonstrated for some viruses by mutant trans-dominant gene products, derived from the movement protein and replication-associated protein genes. In addition to virus-derived sequences, gene sequences of plant origin have also been used for transgenic resistance, and such resistance can be virus-specific, for instance, R genes isolated from resistant plant genotypes, or nonspecific, for example, ribosome inactivating proteins and proteinase inhibitors. Plantibodies and 2-5A synthetase, a class of proteins of mammalian origin, have also been useful in engineering plant virus resistance. In the case of transgenic resistance mediated by viral coat protein, the mechanism of resistance was suggested to operate during the early events of virus infection. However, transgene-mediated RNA silencing and generation of small interfering RNAs appears to be the primary mechanism that confers resistance to plant viruses. Despite the advantages of transgene-mediated resistance, current interest in the development and use of transgenic virus resistant plants is low in most parts of the world. However, because of its real potential, we believe that this technology will have more widespread and renewed interest in the near future. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 17172755 [PubMed - indexed for MEDLINE] 566: Plant Biotechnol J. 2004 Jan;2(1):27-35. NMR profiling of transgenic peas. Charlton A, Allnutt T, Holmes S, Chisholm J, Bean S, Ellis N, Mullineaux P, Oehlschlager S. Department for Environment, Food and Rural Affairs, Central Science Laboratory, Sand Hutton, York, YO41 1LZ, UK. adrian.charlton@csl.gov.uk A high throughput proton nuclear magnetic resonance spectroscopy method for the metabolite fingerprinting of plants was applied to genetically modified peas (Pisum sativum) to determine whether biochemical changes, so called 'unintended effects', beyond those intended by incorporation of a transgene, were detectable. Multivariate analysis of 1H NMR (nuclear magnetic resonance) spectra obtained from uniformly grown glasshouse plants revealed differences between the transgenic and control group that exceeded the natural variation of the plants. When a larger data set of six related transgenic lines was analysed, including a null segregant in addition to the wild-type control, multivariate analysis showed that the distribution of metabolites in the transgenics was different from that of the null segregant. However, the profile obtained from the wild-type material was diverse in comparison with both the transgenics and the null segregant, suggesting that the primary cause of the observed differences was that the transformation process selects for a subset of individuals able to undergo the transformation and selection procedures, and that their descendants have a restricted variation in metabolite profile, rather than that the presence of the transgene itself generates these differences. PMID: 17166140 [PubMed] 567: Plant Biotechnol J. 2003 Sep;1(5):371-80. Presence of potential allergy-related linear epitopes in novel proteins from conventional crops and the implication for the safety assessment of these crops with respect to the current testing of genetically modified crops. Kleter GA, Peijnenburg AA. RIKILT Institute of Food Safety, PO Box 230, NL 6700 AE Wageningen, The Netherlands. Mitochondria of cytoplasmic male sterile crop plants contain novel, chimeric open reading frames. In addition, a number of crops carry endogenous double-stranded ribonucleic acid (dsRNA). In this study, the novel proteins encoded by these genetic components were screened for the presence of potential binding sites (epitopes) of allergy-associated IgE antibodies, as was previously done with transgenic proteins from genetically modified crops. The procedure entails the identification of stretches of at least six contiguous amino acids that are shared by novel proteins and known allergenic proteins. These stretches are further checked for potential linear IgE-binding epitopes. Of the 16 novel protein sequences screened in this study, nine contained stretches of six or seven amino acids that were also present in allergenic proteins. Four cases of similarity are of special interest, given the predicted antigenicity of the identical stretch within the allergenic and novel protein, the IgE-binding by a peptide containing an identical stretch reported in literature, or the multiple incidence of identical stretches of the same allergen within a novel protein. These selected stretches are present in novel proteins derived from oilseed rape and radish (ORF138), rice (dsRNA), and fava bean (dsRNA), and warrant further clinical testing. The frequency of positive outcomes and the sizes of the identical stretches were comparable to those previously found for transgenic proteins in genetically modified crops. It is discussed whether novel proteins from conventional crops should be subject to an assessment of potential allergenicity, a procedure which is currently mandatory for transgenic proteins from genetically modified crops. PMID: 17166136 [PubMed] 568: Behav Brain Res. 2007 Feb 12;177(1):22-9. Epub 2006 Dec 8. Spatial learning in Long-Evans Hooded rats and C57BL/6J mice: different strategies for different performance. Cressant A, Besson M, Suarez S, Cormier A, Granon S. Laboratoire de Physiologie de la Perception et de l'Action, UMR CNRS 7124, Collège de France, Paris, France. Spatial learning abilities of rodents have been extensively used to explore the management of a wide range of cognitive and emotional processes such as learning, memory, attention and anxiety. Knowledge about the organization and processing of spatial learning has mainly been obtained in rats. Due to increasing generation of genetically modified mice, cognitive abilities of mice are now extensively tested. The present paper aimed at comparing spatial representation, learning and strategies in C57BL/6J mice and Long-Evans Hooded rats when subjected to the same spatial learning paradigm, i.e. learning a food location in a crossmaze. We also analyzed the influence of environmental richness on learning modalities in both species. Our results showed that rats and mice could exhibit similar spatial learning abilities in some circumstances. However, Long-Evans rats and C57BL/6J mice may set up different strategies depending on the availability of visual information within the environment. Rats' learning strategies mainly relied on distant visual cues and seemed more efficient than those used by mice as they needed less time than mice to solve the task. We emphasize that the strategies of mice are less robust and flexible than the ones set up by rats. Finally, the richness of the environment was shown to affect speed and quality of spatial learning in both species. Publication Types: Comparative Study Research Support, Non-U.S. Gov't PMID: 17157932 [PubMed - indexed for MEDLINE] 569: Metab Eng. 2007 Jan;9(1):95-111. Epub 2006 Oct 4. Novel transgenic rice overexpressing anthocyanidin synthase accumulates a mixture of flavonoids leading to an increased antioxidant potential. Reddy AM, Reddy VS, Scheffler BE, Wienand U, Reddy AR. Plant Molecular Genetics and Functional Genomics Laboratory, Department of Plant Sciences, School of Life Sciences, University of Hyderabad, AP 500 046, India. a_mmreddy@yahoo.co.in In addition to their plant-associated functions, flavonoids act as antioxidants against harmful free radicals in animals. Genetic engineering of food crops for a mix of antioxidant flavonoids is highly beneficial in promoting human health. Anthocyanidin synthase (ANS) is one of the four dioxygenases (DOX) of the flavonoid biosynthetic pathway that catalyzes the formation of anthocyanidins from leucoanthocyanidins. To investigate whether ANS mediates different DOX reactions of the pathway and produces a mix of flavonoids, the rice ANS cDNA was cloned and overexpressed in a rice mutant Nootripathu (NP). This mutant accumulates proanthocyanidins exclusively in pericarp and absolutely no anthocyanins in any tissue. In silico sequence analysis revealed that ANS contains a double-stranded beta helix and shows high sequence similarity with other DOXs of the pathway including flavonol synthase, flavonone 3beta-hydroxylase and flavone synthase I. Bacterially expressed ANS protein converted dihydroquercetin to quercetin and Pro(35S):ANS complemented the maize a2 mutant in producing anthocyanins in aleurone, suggesting that ANS functions as a DOX with different flavonoid substrates. Similarly, transgenic NP plants overexpressing Pro(MAS):ANS channeled the proanthocaynidin precursors to the production of anthocyanins in pericarp. Transgenics showed approximately ten and four-fold increase in the ANS transcripts and enzyme activity, respectively. As a result, these plants showed an increased accumulation of a mixture of flavonoids and anthocyanins, with a concomitant decrease in proanthocyanidins, suggesting that ANS may act directly on different flavonoid substrates of DOX reactions. Thus, overexpression of ANS in a rice mutant resulted in novel transgenic rice with a mixture of flavonoids and an enhanced antioxidant potential. Publication Types: Research Support, Non-U.S. Gov't PMID: 17157544 [PubMed - indexed for MEDLINE] 570: FEBS Lett. 2006 Dec 22;580(30):6891-7. Epub 2006 Nov 29. Transport of antimony salts by Arabidopsis thaliana protoplasts over-expressing the human multidrug resistance-associated protein 1 (MRP1/ABCC1). Gayet L, Picault N, Cazalé AC, Beyly A, Lucas P, Jacquet H, Suso HP, Vavasseur A, Peltier G, Forestier C. CEA Cadarache, DSV-DEVM--LEMS, UMR 6191 CNRS-CEA-Université Aix-Marseille II, 13108 St Paul lez Durance, France. ABC transporters from the multidrug resistance-associated protein (MRP) subfamily are glutathione S-conjugate pumps exhibiting a broad substrate specificity illustrated by numerous xenobiotics, such as anticancer drugs, herbicides, pesticides and heavy metals. The engineering of MRP transporters into plants might be interesting either to reduce the quantity of xenobiotics taken up by the plant in the context of "safe-food" strategies or, conversely, in the development of phytoremediation strategies in which xenobiotics are sequestered in the vacuolar compartment. In this report, we obtained Arabidopsis transgenic plants overexpressing human MRP1. In these plants, expression of MRP1 did not increase plant resistance to antimony salts (Sb(III)), a classical glutathione-conjugate substrate of MRP1. However, the transporter was fully translated in roots and shoots, and targeted to the plasma membrane. In order to investigate the functionality of MRP1 in Arabidopsis, mesophyll cell protoplasts (MCPs) were isolated from transgenic plants and transport activities were measured by using calcein or Sb(III) as substrates. Expression of MRP1 at the plasma membrane was correlated with an increase in the MCPs resistance to Sb(III) and a limitation of the metalloid content in the protoplasts due to an improvement in Sb(III) efflux. Moreover, Sb(III) transport was sensitive to classical inhibitors of the human MRP1, such as MK571 or glibenclamide. These results demonstrate that a human ABC transporter can be functionally introduced in Arabidopsis, which might be useful, with the help of stronger promoters, to reduce the accumulation of xenobiotics in plants, such as heavy metals from multi-contaminated soils. Publication Types: Research Support, Non-U.S. Gov't PMID: 17150215 [PubMed - indexed for MEDLINE] 571: Plant Biotechnol J. 2005 Nov;3(6):571-82. Engineering a root-specific, repressor-operator gene complex. Kim T, Balish RS, Heaton AC, McKinney EC, Dhankher OP, Meagher RB. Department of Genetics, University of Georgia, Athens, GA 30602, USA. Strong, tissue-specific and genetically regulated expression systems are essential tools in plant biotechnology. An expression system tool called a 'repressor-operator gene complex' (ROC) has diverse applications in plant biotechnology fields including phytoremediation, disease resistance, plant nutrition, food safety, and hybrid seed production. To test this concept, we assembled a root-specific ROC using a strategy that could be used to construct almost any gene expression pattern. When a modified E. coli lac repressor with a nuclear localization signal was expressed from a rubisco small subunit expression vector, S1pt::lacIn, LacIn protein was localized to the nuclei of leaf and stem cells, but not to root cells. A LacIn repressible Arabidopsis actin expression vector A2pot was assembled containing upstream bacterial lacO operator sequences, and it was tested for organ and tissue specificity using beta-glucuronidase (GUS) and mercuric ion reductase (merA) gene reporters. Strong GUS enzyme expression was restricted to root tissues of A2pot::GUS/S1pt::lacIn ROC plants, while GUS activity was high in all vegetative tissues of plants lacking the repressor. Repression of shoot GUS expression exceeded 99.9% with no evidence of root repression, among a large percentage of doubly transformed plants. Similarly, MerA was strongly expressed in the roots, but not the shoots of A2pot::merA/S1pt::lacIn plants, while MerA levels remained high in both shoots and roots of plants lacking repressor. Plants with MerA expression restricted to roots were approximately as tolerant to ionic mercury as plants constitutively expressing MerA in roots and shoots. The superiority of this ROC over the previously described root-specific tobacco RB7 promoter is demonstrated. PMID: 17147628 [PubMed] 572: An Acad Bras Cienc. 2006 Dec;78(4):667-86. GMOs: building the future on the basis of past experience. Reis LF, Van Sluys MA, Garratt RC, Pereira HM, Teixeira MM. Ludwig Institute for Cancer Research, São Paulo, SP, Brazil. lreis@ludwig.org.br Biosafety of genetically modified organisms (GMOs) and their derivatives is still a major topic in the agenda of government and societies worldwide. The aim of this review is to bring into light that data that supported the decision taken back in 1998 as an exercise to stimulate criticism from the scientific community for upcoming discussions and to avoid emotional and senseless arguments that could jeopardize future development in the field. It must be emphasized that Roundup Ready soybean is just one example of how biotechnology can bring in significant advances for society, not only through increased productivity, but also with beneficial environmental impact, thereby allowing more rational use of agricultural pesticides for improvement of the soil conditions. The adoption of agricultural practices with higher yield will also allow better distribution of income among small farmers. New species of genetically modified plants will soon be available and society should be capable of making decisions in an objective and well-informed manner, through collegiate bodies that are qualified in all aspects of biosafety and environmental impact. Publication Types: Review PMID: 17143405 [PubMed - indexed for MEDLINE] 573: Mycologia. 2006 Jul-Aug;98(4):593-7. An optimized method for mycelial compatibility testing in Sclerotinia sclerotiorum. Schafer MR, Kohn LM. Biology Department, University of Toronto at Mississauga, 3359 Mississauga Road North, Mississauga, ON, L5L 1C6, Canada. Classification of isolates into mycelial compatibility groups (MCGs) is used routinely in many laboratories as a quick marker for genotyping Sclerotinia sclerotiorum within populations. Scoring each new sample requires optimization of standardized conditions to support adequate growth of all paired isolates. Appropriate conditions for growth are especially important because diverse compatibility reactions are difficult to categorize and score (e.g., in samples from populations with high genetic diversity, such as those that receive immigration from genetically diverse sources or those that deviate from strict clonality). The current standard medium for MCG testing can be inhibitory to isolates from some samples, confounding scoring of compatibility. We identified two foci for optimization: (i) choice of medium, in this experiment, Patterson's medium amended with red food coloring (termed modified Patterson's medium, MPM, the current standard medium) versus potato dextrose agar (PDA) and (ii) amount of McCormick's red food coloring amended to the growth medium. The red food coloring often yields a red reaction line in incompatible interactions; alternative incompatible reactions are a line of thick or thin hyphae. Based on results to date, self-self pairings of S. sclerotiorum are compatible and are a reliable standard for scoring compatible self-nonself mycelial interactions. PDA amended with 75 microl/L of McCormick's red food coloring was identified as optimal for isolates inhibited by MPM from a highly diverse, recombining population sample. This precisely amended PDA was also suitable for isolates from highly clonal populations that were not inhibited by MPM or by higher concentrations of red food coloring. Under the optimized, standardized conditions all paired isolates grew together and produced interactions that could be scored in repeatedly identifiable categories, compatible or incompatible. Workers are advised to optimize conditions before screening a new population sample. Publication Types: Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. PMID: 17139852 [PubMed - indexed for MEDLINE] 574: Mar Biotechnol (NY). 2007 Jan-Feb;9(1):92-100. Epub 2006 Nov 30. Expression of masu salmon delta5-desaturase-like gene elevated EPA and DHA biosynthesis in zebrafish. Alimuddin, Yoshizaki G, Kiron V, Satoh S, Takeuchi T. Department of Marine Biosciences, Tokyo University of Marine Science and Technology, Minato, Tokyo 108-8477, Japan. Farmed fish could substitute for marine capture fish as a source of fatty acids such as eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) beneficial for human health; however, they require these compounds in their diets. In the present study on a model fish species, we modified the EPA/DHA biosynthesis pathway by overexpression of masu salmon Delta5-desaturase-like gene in zebrafish to increase its ability to synthesize EPA and DHA. Expression of this gene in transgenic fish fed a commercial diet and Artemia helped to improve their EPA content by 1.21-fold and DHA by 1.24-fold. In similar fish that were fed only Artemia the increments were 1.14-fold for EPA and 1.13-fold for DHA, compared with nontransgenic fish. In contrast, eicosatetraenoic acid content decreased, as it is a substrate of Delta5-desaturase, while the total lipid remained constant. The results demonstrated that masu salmon Delta5-desaturase is functional in zebrafish and can modify its fatty acid metabolic pathway. The technique could be applied to farmed fish to generate a nutritionally richer product for human consumption. Publication Types: Research Support, Non-U.S. Gov't PMID: 17136489 [PubMed - indexed for MEDLINE] 575: Ann N Y Acad Sci. 2006 Oct;1081:1-16. Biodiversity and emerging diseases. Maillard JC, Gonzalez JP. Cirad-Emvt/PRISE Hanoi, Vietnam. maillard@fpt.vn First we remind general considerations concerning biodiversity on earth and particularly the loss of genetic biodiversity that seems irreversible whether its origin is directly or indirectly linked to human activities. Urgent and considerable efforts must be made from now on to cataloge, understand, preserve, and enhance the value of biodiversity while ensuring food safety and human and animal health. Ambitious integrated and multifield research programs must be implemented in order to understand the causes and anticipate the consequences of loss of biodiversity. Such losses are a serious threat to sustainable development and to the quality of life of future generations. They have an influence on the natural balance of global biodiversity in particularly in reducing the capability of species to adapt rapidly by genetic mutations to survive in modified ecosystems. Usually, the natural immune systems of mammals (both human and animal), are highly polymorphic and able to adapt rapidly to new situations. We more specifically discuss the fact that if the genetic diversity of the affected populations is low the invading microorganisms, will suddenly expand and create epidemic outbreaks with risks of pandemic. So biodiversity appears to function as an important barrier (buffer), especially against disease-causing organisms, which can function in different ways. Finally, we discuss the importance of preserving biodiversity mainly in the wildlife ecosystems as an integrated and sustainable approach among others in order to prevent and control the emergence or reemergence of diseases in animals and humans (zoonosis). Although plants are also part of this paradigm, they fall outside our field of study. Publication Types: Review PMID: 17135490 [PubMed - indexed for MEDLINE] 576: Biotechnol J. 2006 Dec;1(12):1433-4. Consumer acceptance of ingenic foods. Lusk JL, Rozan A. Department of Agricultural Economics, Oklahoma State University, Stillwater, OK, USA. jayson.lusk@okstate.edu Recent advances in plant molecular biology offer a means of reaping the benefits of biotechnology, while potentially assuaging consumer concerns by re-inserting native DNA back into plants. Results are presented from nationwide surveys in the U.S. and France, indicating that more consumers would accept ingenic plants than transgenic plants, with twice as many U.S. than French consumers considering food produced through biotechnology eatable. PMID: 17124706 [PubMed - indexed for MEDLINE] 577: Science. 2006 Nov 24;314(5803):1298-301. Comment in: Science. 2006 Nov 24;314(5803):1252-3. A NAC Gene regulating senescence improves grain protein, zinc, and iron content in wheat. Uauy C, Distelfeld A, Fahima T, Blechl A, Dubcovsky J. Department of Plant Sciences, University of California, One Shields Avenue, Davis, CA 95616, USA. Enhancing the nutritional value of food crops is a means of improving human nutrition and health. We report here the positional cloning of Gpc-B1, a wheat quantitative trait locus associated with increased grain protein, zinc, and iron content. The ancestral wild wheat allele encodes a NAC transcription factor (NAM-B1) that accelerates senescence and increases nutrient remobilization from leaves to developing grains, whereas modern wheat varieties carry a nonfunctional NAM-B1 allele. Reduction in RNA levels of the multiple NAM homologs by RNA interference delayed senescence by more than 3 weeks and reduced wheat grain protein, zinc, and iron content by more than 30%. Publication Types: Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. PMID: 17124321 [PubMed - indexed for MEDLINE] 578: Planta. 2007 Apr;225(5):1165-78. Epub 2006 Nov 21. Metabolic diversion of the phenylpropanoid pathway causes cell wall and morphological changes in transgenic tobacco stems. Merali Z, Mayer MJ, Parker ML, Michael AJ, Smith AC, Waldron KW. Institute of Food Research, , Norwich Research Park, Colney, Norwich, NR4 7UA, UK. Studies involving transgenic plants with modifications in the lignin pathway reported to date, have received a relatively preliminary characterisation in relation to the impact on vascular integrity, biomechanical properties of tissues and carbon allocation to phenolic pools. Therefore, in this study transgenic tobacco plants (Nicotiana tabacum cv XHFD 8) expressing various levels of a bacterial 4-hydroxycinnamoyl-CoA hydratase/lyase (HCHL) gene have been characterised for cell wall and related morphological changes. The HCHL enzyme converts p-coumaroyl-CoA to 4-hydroxybenzaldehyde thereby rerouting the phenylpropanoid pathway. Plants expressing high levels of HCHL activity exhibited reduced lignin deposition, impaired monolignol biosynthesis and vascular integrity. The plants also exhibited reduction in stem toughness concomitant with a massive reduction in both the cell wall esterified and soluble phenolics. A notable result of redirecting the carbon flux was the wall-bound accretion of vanillin and vanillic acid, probably due to the shunt pathway. Intracellular accumulation of novel metabolites such as hydroxybenzoic and vanillic acid derivatives also occurred in the transgenic plants. A line with intermediate levels of HCHL expression conferred correspondingly reduced lignin deposition, toughness and phenolics. This line displayed a normal morphology but distorted vasculature. Coloration of the xylem has been previously attributed to incorporation of alternative phenolics, whereas results from this study indicate that the coloration is likely to be due to the association of low molecular weight phenolics. There was no evidence of increased growth or enhanced cellulose biosynthesis as a result of HCHL expression. Hence, rerouting the phenylpropanoid biosynthetic pathway quantitatively and qualitatively modifies cell wall-bound phenolics and vascular structure. Publication Types: Research Support, Non-U.S. Gov't PMID: 17120022 [PubMed - indexed for MEDLINE] 579: J Agric Food Chem. 2006 Nov 29;54(24):8984-94. Applications of metabolomics in agriculture. Dixon RA, Gang DR, Charlton AJ, Fiehn O, Kuiper HA, Reynolds TL, Tjeerdema RS, Jeffery EH, German JB, Ridley WP, Seiber JN. Plant Biology Division, Samuel Roberts Noble Foundation, 2510 Sam Noble Parkway, Ardmore, OK 73401, USA. Biological systems are exceedingly complex. The unraveling of the genome in plants and humans revealed fewer than the anticipated number of genes. Therefore, other processes such as the regulation of gene expression, the action of gene products, and the metabolic networks resulting from catalytic proteins must make fundamental contributions to the remarkable diversity inherent in living systems. Metabolomics is a relatively new approach aimed at improved understanding of these metabolic networks and the subsequent biochemical composition of plants and other biological organisms. Analytical tools within metabolomics including mass spectrometry (MS) and nuclear magnetic resonance (NMR) spectroscopy can profile the impact of time, stress, nutritional status, and environmental perturbation on hundreds of metabolites simultaneously resulting in massive, complex data sets. This information, in combination with transcriptomics and proteomics, has the potential to generate a more complete picture of the composition of food and feed products, to optimize crop trait development, and to enhance diet and health. Selected presentations from an American Chemical Society symposium held in March 2005 have been assembled to highlight the emerging application of metabolomics in agriculture. Publication Types: Review PMID: 17117782 [PubMed - indexed for MEDLINE] 580: Proc Natl Acad Sci U S A. 2006 Nov 28;103(48):18054-9. Epub 2006 Nov 16. Engineering cottonseed for use in human nutrition by tissue-specific reduction of toxic gossypol. Sunilkumar G, Campbell LM, Puckhaber L, Stipanovic RD, Rathore KS. Institute for Plant Genomics and Biotechnology and Department of Soil and Crop Sciences, Texas A&M University, College Station, TX 77843, USA. Global cottonseed production can potentially provide the protein requirements for half a billion people per year; however, it is woefully underutilized because of the presence of toxic gossypol within seed glands. Therefore, elimination of gossypol from cottonseed has been a long-standing goal of geneticists. Attempts were made to meet this objective by developing so-called "glandless cotton" in the 1950s by conventional breeding techniques; however, the glandless varieties were commercially unviable because of the increased susceptibility of the plant to insect pests due to the systemic absence of glands that contain gossypol and other protective terpenoids. Thus, the promise of cottonseed in contributing to the food requirements of the burgeoning world population remained unfulfilled. We have successfully used RNAi to disrupt gossypol biosynthesis in cottonseed tissue by interfering with the expression of the delta-cadinene synthase gene during seed development. We demonstrate that it is possible to significantly reduce cottonseed-gossypol levels in a stable and heritable manner. Results from enzyme activity and molecular analyses on developing transgenic embryos were consistent with the observed phenotype in the mature seeds. Most relevant, the levels of gossypol and related terpenoids in the foliage and floral parts were not diminished, and thus their potential function in plant defense against insects and diseases remained untouched. These results illustrate that a targeted genetic modification, applied to an underutilized agricultural byproduct, provides a mechanism to open up a new source of nutrition for hundreds of millions of people. Publication Types: Research Support, Non-U.S. Gov't PMID: 17110445 [PubMed - indexed for MEDLINE] 581: Transgenic Res. 2007 Apr;16(2):239-49. Epub 2006 Nov 15. Erratum in: Transgenic Res. 2007 Apr;16(2):251. Elastin-like polypeptide fusions enhance the accumulation of recombinant proteins in tobacco leaves. Patel J, Zhu H, Menassa R, Gyenis L, Richman A, Brandle J. Agriculture and AgriFood Canada, Southern Crop Protection and Food Research Center, 1391 Sandford Street, London, Ontario, Canada, N5V 4T3. The production of recombinant proteins in plants is an active area of research and many different high-value proteins have now been produced in plants. Tobacco leaves have many advantages for recombinant protein production particularly since they allow field production without seeds, flowers or pollen and therefore provide for contained production. Despite these biosafety advantages recombinant protein accumulation in leaves still needs to be improved. Elastin-like polypeptides are repeats of the amino acids "VPGXG" that undergo a temperature dependant phase transition and have utility in the purification of recombinant proteins but can also enhance the accumulation of recombinant proteins they are fused to. We have used a 11.3 kDa elastin-like polypeptide as a fusion partner for three different target proteins, human interleukin-10, murine interleukin-4 and the native major ampullate spidroin protein 2 gene from the spider Nephila clavipes. In both transient analyses and stable transformants the concentrations of the fusion proteins were at least an order of magnitude higher for all of the fusion proteins when compared to the target protein alone. Therefore, fusions with a small ELP tag can be used to significantly enhance the accumulation of a range of different recombinant proteins in plant leaves. PMID: 17106768 [PubMed - indexed for MEDLINE] 582: Transgenic Res. 2007 Feb;16(1):51-63. Epub 2006 Nov 15. Sources of uncertainty in the quantification of genetically modified oilseed rape contamination in seed lots. Begg GS, Cullen DW, Iannetta PP, Squire GR. Scottish Crop Research Institute, Invergowrie, Dundee, DD2 5DA, UK. gbegg@scri.ac.uk Testing of seed and grain lots is essential in the enforcement of GM labelling legislation and needs reliable procedures for which associated errors have been identified and minimised. In this paper we consider the testing of oilseed rape seed lots obtained from the harvest of a non-GM crop known to be contaminated by volunteer plants from a GM herbicide tolerant variety. The objective was to identify and quantify the error associated with the testing of these lots from the initial sampling to completion of the real-time PCR assay with which the level of GM contamination was quantified. The results showed that, under the controlled conditions of a single laboratory, the error associated with the real-time PCR assay to be negligible in comparison with sampling error, which was exacerbated by heterogeneity in the distribution of GM seeds, most notably at a small scale, i.e. 25 cm3. Sampling error was reduced by one to two thirds on the application of appropriate homogenisation procedures. Publication Types: Research Support, Non-U.S. Gov't PMID: 17106767 [PubMed - indexed for MEDLINE] 583: Transgenic Res. 2007 Aug;16(4):437-48. Epub 2006 Nov 14. A leaf-based regeneration and transformation system for maize (Zea mays L.). Ahmadabadi M, Ruf S, Bock R. Max-Planck-Institut für Molekulare Pflanzenphysiologie, Am Mühlenberg 1, D-14476, Potsdam-Golm, Germany. Efficient methods for in vitro propagation, regeneration, and transformation of plants are of pivotal importance to both basic and applied research. While being the world's major food crops, cereals are among the most difficult-to-handle plants in tissue culture which severely limits genetic engineering approaches. In maize, immature zygotic embryos provide the predominantly used material for establishing regeneration-competent cell or callus cultures for genetic transformation experiments. The procedures involved are demanding, laborious and time consuming and depend on greenhouse facilities. We have developed a novel tissue culture and plant regeneration system that uses maize leaf tissue and thus is independent of zygotic embryos and greenhouse facilities. We report here: (i) a protocol for the efficient induction of regeneration-competent callus from maize leaves in the dark, (ii) a protocol for inducing highly regenerable callus in the light, and (iii) the use of leaf-derived callus for the generation of stably transformed maize plants. Publication Types: Research Support, Non-U.S. Gov't PMID: 17103238 [PubMed - indexed for MEDLINE] 584: Transgenic Res. 2007 Feb;16(1):109-13. Epub 2006 Nov 11. Antioxidant value addition in human diets: genetic transformation of Brassica juncea with gamma-TMT gene for increased alpha-tocopherol content. Yusuf MA, Sarin NB. School of Life Sciences, Jawaharlal Nehru University, New Delhi, 110067, India. Alpha-tocopherol, the most biologically active form of vitamin E, is implicated in decreasing the risk of several types of cancers, coronary heart disease and a number of degenerative human conditions, when taken in excess of the recommended daily allowance. Natural alpha-tocopherol has twice the bioavailability of the synthetic isomer. This study describes a successful attempt at fortifying human diets with natural alpha-tocopherol by taking recourse to genetic engineering of an important oilseed crop, Brassica juncea. Gamma-tocopherol methyl transferase cDNA from Arabidopsis thaliana, coding for the enzyme catalysing the conversion of the large gamma-tocopherol pool to alpha-tocopherol, was overexpressed in B. juncea plants. The successful integration of the transgene was confirmed by PCR and Southern blot analysis, while the enhanced transcript level was evident in the northern blot analysis. HPLC analysis of the seeds of the T1 transgenic lines showed a shift in tocopherol profile with the highest over-expressors having alpha-tocopherol levels as high as sixfold over the non-transgenic controls. This study discusses the production of a transgenic oilseed crop with high alpha-tocopherol levels, which can provide a feasible, innocuous, and inexpensive way of taking the beneficial effects of high alpha-tocopherol intake to the masses. Publication Types: Research Support, Non-U.S. Gov't PMID: 17103027 [PubMed - indexed for MEDLINE] 585: Theriogenology. 2007 Jan 1;67(1):188-97. Epub 2006 Nov 13. Current status of regulating biotechnology-derived animals in Canada: animal health and food safety considerations. Kochhar HP, Evans BR. Canadian Food Inspection Agency, Ottawa, Ont, Canada. hkochhar@inspection.gc.ca Development of an effective regulatory system for genetically engineered animals and their products has been the subject of increasing discussion among researchers, industry and policy developers, as well as the public. Since transgenesis and cloning are relatively new scientific techniques, transgenic animals are 'novel' organisms for which there is limited information. The issues associated with the regulation of transgenic animals pertain to environmental impact, human food safety, animal health and welfare, trade and ethics. It is a challenge for the developers to prove the safety of the products of biotechnology-derived animals and also for regulators to regulate this increasingly powerful technology with limited background information. In principle, an effective regulatory sieve should permit safe products while forming a formidable barrier for those posing an unacceptable risk. Regulatory initiatives for biotechnology-derived animals and their products should be able to ensure high standards for human and animal health, a sound scientific basis for evaluation; transparency and public involvement, and maintenance of genetic diversity. This review proposes a regulatory regime that is based on scientific risk based assessment and approval of products or by-products of biotechnology-derived animals and its application in context to Canadian regulations. Publication Types: Review PMID: 17097725 [PubMed - indexed for MEDLINE] 586: Nat Biotechnol. 2006 Nov;24(11):1329; author reply 1331-3. Comment on: Nat Biotechnol. 2006 Jul;24(7):753. 'Cisgenic' as a product designation. Giddings LV. Publication Types: Comment Letter PMID: 17093471 [PubMed - indexed for MEDLINE] 587: Nat Biotechnol. 2006 Nov;24(11):1329-31; author reply 1331-3. Comment on: Nat Biotechnol. 2006 Jul;24(7):753. 'Cisgenic' as a product designation. de Cock Buning T, Lammerts van Bueren ET, Haring MA, de Vriend HC, Struik PC. Publication Types: Comment Letter PMID: 17093470 [PubMed - indexed for MEDLINE] 588: Nat Biotechnol. 2006 Nov;24(11):1327-9; author reply 1331-3. Comment on: Nat Biotechnol. 2006 Jul;24(7):753. 'Cisgenic' as a product designation. Schubert D, Williams D. Publication Types: Comment Letter PMID: 17093469 [PubMed - indexed for MEDLINE] 589: Nat Biotechnol. 2006 Nov;24(11):1301-2. Liberty Link rice raises specter of tightened regulations. Vermij P. Publication Types: News PMID: 17093456 [PubMed - indexed for MEDLINE] 590: Nature. 2006 Nov 9;444(7116):137. A breed apart. [No authors listed] PMID: 17093424 [PubMed - indexed for MEDLINE] 591: J Allergy Clin Immunol. 2006 Nov;118(5):1176-83. Epub 2006 Sep 8. Reduced allergenicity of tomato fruits harvested from Lyc e 1-silenced transgenic tomato plants. Le LQ, Mahler V, Lorenz Y, Scheurer S, Biemelt S, Vieths S, Sonnewald U. Department of Biochemistry, Friedrich-Alexander University Erlangen-Nuremberg, Erlangen, Germany. BACKGROUND: Profilin is a small actin-binding protein that contributes to the allergenic potency of many fruits and vegetables, including tomato. Two highly similar genes encoding tomato profilin have been isolated and designated as allergen Lyc e 1.01 and Lyc e 1.02. OBJECTIVE: The aim of the study was to generate profilin-reduced hypoallergenic tomato fruits by silencing of both genes in transgenic tomato plants by means of RNA interference (RNAi). METHODS: The efficiency of gene silencing was documented by means of Northern blotting, immunoblotting, and skin prick testing. RESULTS: Quantification of the remaining protein revealed that profilin accumulation in transgenic fruits was decreased 10-fold compared with that seen in untransformed controls. This decrease was sufficient to cause a reduced allergenic reactivity in patients with tomato allergy, as determined with skin prick tests. Because most patients with tomato allergy are not monosensitized to profilin, the IgE reactivity to the profilin-silenced tomato fruits in vivo varied widely between individuals tested. CONCLUSION: We could demonstrate the efficient silencing of both profilin genes in transgenic tomato plants using RNAi. This resulted in Lyc e 1-diminished tomato fruits, providing proof of concept and demonstrating that RNAi can be used to design allergen-reduced food. However, simultaneous silencing of multiple allergens will be required to design hypoallergenic tomatoes. CLINICAL IMPLICATIONS: Our findings demonstrate the feasibility of creating low-allergenic food by using RNAi. This concept constitutes a novel approach to allergen avoidance. Publication Types: Research Support, Non-U.S. Gov't PMID: 17088146 [PubMed - indexed for MEDLINE] 592: J Vet Med Sci. 2006 Oct;68(10):1113-5. Effects of feeding calves genetically modified corn bt11: a clinico-biochemical study. Shimada N, Murata H, Mikami O, Yoshioka M, Guruge KS, Yamanaka N, Nakajima Y, Miyazaki S. Safety Research Team, National Institute of Animal Health, Ibaraki, Japan. Genetically modified corn Bt11 is insect-resistant and expresses Cry1Ab toxin, an insecticidal protein, in kernels. Although Bt11 corn is considered safe based on animal performance, there are no reports available on the clinico-biochemical effects of feeding it to cattle. In this study, we evaluated the effects of feeding Bt11 to calves, using blood and ruminal clinico-biochemical parameters. Our three-month-long feeding experiment demonstrated that calves (n=6), fed with a ration containing 43.3% of Bt11 corn kernels as dry matter, did not develop any discernible clinical, hematological, biochemical, or ruminal abnormalities as compared with control calves (n=6) fed non-Bt11 corn. The results suggest that the transgenic Bt11 has no negative clinico-biochemical effects on calves. Publication Types: Randomized Controlled Trial Research Support, Non-U.S. Gov't PMID: 17085894 [PubMed - indexed for MEDLINE] 593: Biotechnol Lett. 2006 Dec;28(24):1983-91. Epub 2006 Nov 2. Review: Genetically modified plants for the promotion of human health. Yonekura-Sakakibara K, Saito K. RIKEN Plant Science Center, 1-7-22, Suehiro, Tsurumi-ku, Yokohama, 230-0045, Japan. Plants are attractive biological resources because of their ability to produce a huge variety of chemical compounds, and the familiarity of production in even the most rural settings. Genetic engineering gives plants additional characteristics and value for cultivation and post-harvest. Genetically modified (GM) plants of the "first generation" were conferred with traits beneficial to producers, whereas GM plants in subsequent "generations" are intended to provide beneficial traits for consumers. Golden Rice is a promising example of a GM plant in the second generation, and has overcome a number of obstacles for practical use. Furthermore, consumer-acceptable plants with health-promoting properties that are genetically modified using native genes are being developed. The emerging technology of metabolomics will also support the commercial realization of GM plants by providing comprehensive analyzes of plant biochemical components. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 17080241 [PubMed - indexed for MEDLINE] 594: Med Law. 2006 Sep;25(3):491-502. Biotechnology entrepreneurship and ethics: principles, paradigms, and products. Kuszler PC. University of Washington School of Law, William H Gates Hall, Seattle, USA. Biotechnology, whether in the context of new drugs derived from DNA and genetic technology, genetically modified food, or biologics making use of living cells, raises ethical concerns at a variety of different levels. At the research level, there is concern that the very nature of research is being subverted, rather than enhanced, by entrepreneurship. This area of ethical concern has intensified in the United States as a result of the conflicts of interests resulting from the growing alliance between University academia and private industry in the research enterprise. As we travel down the research path into development of a drug or technology, ethical questions arise with respect to protecting human subjects and society from danger and exploitation by researchers. As development gives way to marketing and dissemination of a new product, government regulators are pressed to get drugs and biologics through the regulatory pipeline into the market faster, walking an ethical tightrope between speed and safety. As new biotechnology products enter the market place, doctors and patients traverse yet another tightrope, that between unknown risk and the promise of benefit. And finally, patent protection is increasingly viewed as a unethical culprit in keeping prices high and depriving the global poor from lifesaving drugs and biologics. Bioethics has, to date, been largely a creation of Western research and medicine. As such it is wholly inadequate to respond to the cascade of ethical issues that flow from a vibrant biotechnology industry. And if biotechnology is in its infancy, as most believe, it is crucial that scientists, entrepreneurs and governments engage in dialogue about the ethical and societal questions raised on the road of scientific progress. PMID: 17078522 [PubMed - indexed for MEDLINE] 595: Gig Sanit. 2006 Jul-Aug;(4):7-11. [Hygienic characteristics of foodstuffs containing genetically modified components] [Article in Russian] Beliaev EN, Ivanov AA, Fokin MV. The paper analyzes the results of the investigations of raw foods, foodstuffs for genetically modified components, conducted by the state sanitary and epidemiological service of the Russian Federation during its current sanitary inspection. The presented materials cover 2003-2004. The findings suggest that there is a great deal of foods containing genetically modified sources on the market and show the priority groups of foodstuffs and the distribution of these foods on the territory of the Russian Federation. Publication Types: English Abstract PMID: 17078283 [PubMed - indexed for MEDLINE] 596: Theriogenology. 2007 Jan 1;67(1):1-206. Epub 2006 Oct 27. Proceedings of the IETS (International Embryo Transfer Society) Pre-Conference Symposia, Kyoto, Japan, 6 January 2007. [No authors listed] Publication Types: Congresses Overall PMID: 17069880 [PubMed - indexed for MEDLINE] 597: Annu Rev Plant Biol. 2007;58:1-19. From analysis of mutants to genetic engineering. von Wettstein D. Department of Crop and Soil Sciences, School of Molecular Biosciences and Center for Integrated Biotechnology, Washington State University, Pullman, WA 99164-6420, USA. diter@wsu.edu This chapter describes the research of developing transgenic barley for synthesis of recombinant proteins with practical significance and of metabolic engineering of proanthocyanidin-free barley. The results were obtained by graduate students, postdoctoral researchers, and visiting scientists at the Carlsberg Laboratory from 1972-1996 and during the past ten years at Washington State University. It is written in appreciation of their enthusiasm, skill, and perseverance. Publication Types: Review PMID: 17067283 [PubMed - indexed for MEDLINE] 598: J Biotechnol. 2007 Jan 30;128(1):194-203. Epub 2006 Sep 23. Effect of storage and processing on plasmid, yeast and plant genomic DNA stability in juice from genetically modified oranges. Weiss J, Ros-Chumillas M, Peña L, Egea-Cortines M. Agricultural Science and Technology Department, Genetics, Universidad Politécnica de Cartagena, 30203 Cartagena, Spain. julia.weiss@upct.es Recombinant DNA technology is an important tool in the development of plant varieties with new favourable features. There is strong opposition towards this technology due to the potential risk of horizontal gene transfer between genetically modified plant material and food-associated bacteria, especially if genes for antibiotic resistance are involved. Since horizontal transfer efficiency depends on size and length of homologous sequences, we investigated the effect of conditions required for orange juice processing on the stability of DNA from three different origins: plasmid DNA, yeast genomic DNA and endogenous genomic DNA from transgenic sweet orange (C. sinensis L. Osb.). Acidic orange juice matrix had a strong degrading effect on plasmid DNA which becomes apparent in a conformation change from supercoiled structure to nicked, linear structure within 5h of storage at 4 degrees C. Genomic yeast DNA was degraded during exposure to acidic orange juice matrix within 4 days, and also the genomic DNA of C. sinensis suffered degradation within 2 days of storage as indicated by amplification results from transgene markers. Standard pasteurization procedures affected DNA integrity depending on the method and time used. Our data show that the current standard industrial procedures to pasteurize orange juice as well as its acidic nature causes a strong degradation of both yeast and endogenous genomic DNA below sizes reported to be suitable for horizontal gene transfer. PMID: 17064805 [PubMed - indexed for MEDLINE] 599: J Agric Food Chem. 2006 Nov 1;54(22):8640-7. Safety assessment of cre recombinase. Hileman RE, Bonner HK, Kaempfe TA, Hammond BG, Glenn KC. Monsanto Company, St. Louis, Missouri 63167, USA. ronald.e.hileman@monsanto.com Cre recombinase, when used as a tool in agricultural biotechnology, can precisely excise DNA sequences that may be useful in the introduction of a new trait but are not needed in the commercial product. Although the cre genetic material would not be present in the final product, the present studies were performed to assess the safety of Cre recombinase to provide confirmatory evidence of the safe use of Cre-lox technology in agricultural biotechnology. Cre recombinase shares no relevant sequence similarity to known allergens or toxins. When Cre recombinase was exposed to a pH 1.2 solution of simulated gastric fluid lacking pepsin, CD spectroscopy showed that there was a loss of secondary structure and that the protein was no longer active in a functional assay. Cre recombinase was degraded rapidly when exposed to pepsin in a standardized gastric digestion model; therefore, Cre recombinase would not survive the harsh gastric environment. When orally administered to mice as an acute dosage of 53 mg/kg of body weight, no treatment-related adverse findings were observed. These data support the conclusion that human and animal dietary exposure to Cre recombinase pose no known safety concerns; consistent with the fact that bacteriophage P1, the source of the cre gene and expressed protein, is commonly encountered in the environment and in normal enteric bacteria without reports of adverse consequences. PMID: 17061845 [PubMed - indexed for MEDLINE] 600: J Agric Environ Ethics. 2006;19(3):253-67. Including public perspectives in industrial biotechnology and the biobased economy. Paula L, Birrer F. Institute of Biology, Biology and Society, Leiden University, PO Box 9516, 2300 RA Leiden, The Netherlands. lepaula@rulsfb.leidenuniv.nl Industrial ("white") biotechnology promises to contribute to a more sustainable future. Compared to current production processes, cases have been identified where industrial biotechnology can decrease the amount of energy and raw materials used to make products and also reduce the amount of emissions and waste produced during production. However, switching from products based on chemical production processes and fossil fuels towards "biobased" products is at present not necessarily economically viable. This is especially true for bulk products, for example ethanol production from biomass. Therefore, scientists are also turning to genetic modification as a means to develop organisms that can produce at lower costs. These include not only micro-organisms, but also organisms used in agriculture for food and feed. The use of genetic modification for "deliberate release" purposes, in particular, has met great opposition in Europe. Many industrial biotechnology applications may, due to their scale, entail deliberate releases of GM organisms. Thus, the biobased economy brings back a familiar question; is it ethically justifiable, and acceptable to citizens, to expose the environment and society to the risks associated with GM, in order to protect that same environment and to sustain our affluent way of life? For a successful innovation towards a biobased economy, its proponents, especially producers, need to take into account (take responsibility for) such issues when developing new products and processes. These issues, and how scientists can interact with citizens about them in a timely way, are further explored in projects at Delft University and Leiden University, also in collaboration with Utrecht University. PMID: 17061382 [PubMed - indexed for MEDLINE] 601: J Agric Environ Ethics. 2006;19(3):239-52. Journalism and science: how to erode the idea of knowledge. Meyer G. The International Center for Business and Politics, Copenhagen Business School, Monrads Alle 7, DK 2500, Valby, Denmark. sprogbrug@get2net.dk This paper discusses aspects of the relationship between the scientific community and the public at large. Inspired by the European public debate on genetically modified crops and food, ethical challenges to the scientific community are highlighted. This is done by a discussion of changes that are likely to occur to journalistic attitudes--mirroring changing attitudes in the wider society--towards science and scientific researchers. Two journalistic conventions--those of science transmission and of investigative journalism--are presented and discussed in relation to the present drive towards commercialization within the world of science: how are journalists from these different schools of thought likely to respond to the trend of commercialization? Likely journalistic reactions could, while maintaining the authority of the scientific method, be expected to undermine public trust in scientists. In the long term, this may lead to an erosion of the idea of knowledge as something that cannot simply be reduced to the outcome of negotiation between stakeholders. It is argued that science is likely to be depicted as a fallen angel. This may be countered, it is posited, by science turning human, by recognizing its membership of society, and by recognizing that such membership entails more than just commercial relations. To rethink its relationship with the public at large--and, in particular, to rethink the ideal of disinterested science--is an ethical challenge facing the scientific community. PMID: 17061381 [PubMed - indexed for MEDLINE] 602: J Agric Environ Ethics. 2006;19(3):225-38. The moral difference between intragenic and transgenic modification of plants. Myskja BK. Department of Philosophy, NTNU Trondheim, NO-7491 Trondheim, Norway. bjorn.myskja@hf.ntnu.no Public policy on the development and use of genetically modified organisms (GMOs) has mainly been concerned with defining proper strategies of risk management. However, surveys and focus group interviews show that although lay people are concerned with risks, they also emphasize that genetic modification is ethically questionable in itself. Many people feel that this technology "tampers with nature" in an unacceptable manner. This is often identified as an objection to the crossing of species borders in producing transgenic organisms. Most scientists reject these opinions as based on insufficient knowledge about biotechnology, the concept of species, and nature in general. Some recent projects of genetic modification aim to accommodate the above mentioned concerns by altering the expression of endogenous genes rather than introducing genes from other species. There can be good scientific reasons for this approach, in addition to strategic reasons related to greater public acceptability. But are there also moral reasons for choosing intragenic rather than transgenic modification? I suggest three interrelated moral reasons for giving priority to intragenic modification. First, we should respect the opinions of lay people even when their view is contrary to scientific consensus; they express an alternative world-view, not scientific ignorance. Second, staying within species borders by strengthening endogenous traits reduces the risks and scientific uncertainty. Third, we should show respect for nature as a complex system of laws and interconnections that we cannot fully control. The main moral reason for intragenic modification, in our view, is the need to respect the "otherness" of nature. PMID: 17061380 [PubMed - indexed for MEDLINE] 603: Dev Biol (Basel). 2006;126:79-86; discussion 324-5. In-house validation and quality control of real-time PCR methods for GMO detection: a practical approach. Ciabatti I, Froiio A, Gatto F, Amaddeo D, Marchesi U. Istituto Zooprofilattico Sperimentale Lazio e Toscana, National Reference Center for GMO Analysis, Rome, Italy. iciabatti@rm.izs.it GMO detection and quantification methods in the EU are mainly based on real-time PCR. The analytical methods in use must be validated, first on an intra-laboratory scale and through a collaborative trial thereafter. Since a consensual protocol for intra-laboratory validation of real-time PCR methods is lacking, we provide a practical approach for the in-house validation of quantitative real-time PCR methods, establishing acceptability criteria and quality controls for PCR runs. Parameters such as limit of detection, limit of quantification, precision, trueness, linear dynamic range, PCR efficiency, robustness and specificity are considered. The protocol is sufficiently detailed to be directly applicable, increases the reliability of results and their harmonization among different laboratories, and represents a necessary preliminary step before proceeding to a time-consuming and costly full validation study. PMID: 17058483 [PubMed - indexed for MEDLINE] 604: Ann N Y Acad Sci. 2006 Aug;1072:176-86. Therapeutic drug delivery by genetically modified Lactococcus lactis. Steidler L, Rottiers P. Alimentary Pharmabiotic Centre, Transgenic Bacteriology, University College Cork, Western Road, Cork, Ireland. l.steidler@ucc.ie Food-grade bacteria have been consumed throughout history without associated pathologies and are, therefore, absolutely safe to ingest. Unexpectedly, Lactococcus lactis (L. lactis), known from cheese production, can be genetically engineered to constantly secrete satisfactory amounts of bioactive cytokines. Both of these features enabled the development of a new kind of topical delivery system: topical and active delivery of therapeutic proteins by genetically modified micro-organisms. The host organism's record inspired the development of applications that target intestinal diseases. In a variety of mouse models, chronic colon inflammation can be successfully treated with (interleukin) IL-10-secreting L. lactis. Trefoil factor (TFF) producer strains have also been shown to be very effective in the treatment of acute colitis. Such novel therapeutic strains are textbook examples of genetically modified (GM) organisms. There are legitimate concerns with regard to the deliberate release of GM micro-organisms. On development of these applications, therefore, we have engineered these bacteria in such a way that biological containment is guaranteed. The essential gene thyA, encoding thymidylate synthase, has been exchanged for IL-10. This makes the GM strain critically dependent on thymidine. Lack of thymidine, for example, resulting from thymidine consumption by thyA-deficient strains-will irreversibly lead to induced "thymidine-less death." This accomplishment has created the possibility of using this strategy for application in human medicine. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 17057198 [PubMed - indexed for MEDLINE] 605: Theriogenology. 2007 Jan 1;67(1):185-7. Epub 2006 Oct 20. Regulatory considerations on transgenic livestock in Japan in relation to the Cartagena protocol. Yamanouchi K. Nippon Institute for Biological Science, 9-2221-1 Shin-Machi, Ome, Tokyo, Japan. yamanokazu@aol.com In Japan, the development and application of living modified organisms (LMOs) are regulated by law (conservation and sustainable use of biological diversity law). Procedures are classed as type 1 for the use of LMOs where no preventive measures against their dispersal into the environment are required and type 2 for the use of LMOs where preventive measures are stipulated. Development and research on transgenic livestock falls under the responsibility of the Ministry of Education, Culture, Science, Sports and Technology. Field use of transgenic livestock is controlled by the Ministry of Agriculture, Forestry and Fisheries. The author describes risk assessment and management of transgenic livestock by both ministries. Publication Types: Review PMID: 17055568 [PubMed - indexed for MEDLINE] 606: Food Chem Toxicol. 2007 Mar;45(3):364-77. Epub 2006 Sep 14. Safety testing of GM-rice expressing PHA-E lectin using a new animal test design. Poulsen M, Schrøder M, Wilcks A, Kroghsbo S, Lindecrona RH, Miller A, Frenzel T, Danier J, Rychlik M, Shu Q, Emami K, Taylor M, Gatehouse A, Engel KH, Knudsen I. Department of Toxicology and Risk Assessment, Danish Institute for Food and Veterinary Research, Mørkhøj Bygade 19, DK-2860 Søborg, Denmark. mop@dfvf.dk The 90-day animal study is the core study for the safety assessment of genetically modified foods in the SAFOTEST project. The model compound tested in the 90-day study was a rice variety expressing the kidney bean Phaseolus vulgaris lectin agglutinin E-form (PHA-E lectin). Female Wistar rats were given a nutritionally balanced purified diet with 60% parental rice, 60% PHA-E rice or 60% PHA-E rice spiked with 0.1% recombinant PHA-E lectin for 90 days. This corresponded to a mean daily PHA-E lectin intake of approximately 0, 30 and 100mg/kg body weight for each group, respectively. The spiking was used to increase the specificity and to demonstrate the sensitivity of the study. A range of biological, biochemical, microbiological and pathological parameters were examined and significant differences in weight of small intestine, stomach and pancreas and plasma biochemistry were seen between groups. Included in this paper are also data from the molecular characterisation and chemical analysis of the PHA-E rice, from the construction and production of the PHA-E lectin, and from the preceding 28-day in vivo study where the toxicity of the pure PHA-E lectin was determined. In conclusion, the combined use of information from the compositional analysis, the 28-day study and the characterisation of the PHA-E rice and the PHA-E lectin has improved the design of the 90-day study. The spiking procedure has facilitated the interpretation of the results of the study and transferred it into a valuable tool for the future safety testing of genetically modified foods. Publication Types: Evaluation Studies Research Support, Non-U.S. Gov't PMID: 17052831 [PubMed - indexed for MEDLINE] 607: Food Chem Toxicol. 2007 Mar;45(3):350-63. Epub 2006 Sep 14. A 90-day safety study in Wistar rats fed genetically modified rice expressing snowdrop lectin Galanthus nivalis (GNA). Poulsen M, Kroghsbo S, Schrøder M, Wilcks A, Jacobsen H, Miller A, Frenzel T, Danier J, Rychlik M, Shu Q, Emami K, Sudhakar D, Gatehouse A, Engel KH, Knudsen I. Department of Toxicology and Risk Assessment, Danish Institute for Food and Veterinary Research, Mørkhøj Bygade 19, DK-2860 Søborg, Denmark. mop@dfvf.dk Genetically modified plants expressing insecticidal traits offer a new strategy for crop protection, but at the same time present a challenge in terms of food safety assessment. The present 90-day feeding study was designed to assess the safety of a rice variety expressing the snowdrop Galanthus nivalis lectin (GNA lectin), and forms part of a EU-funded project where the objective has been to develop and validate sensitive and specific methods to assess the safety of genetically modified foods. Male and female Wistar rats were given a purified diet containing either 60% genetically modified or parental rice for 90 days. This corresponds to a mean daily GNA lectin intake of approximately 58 and 67mg/kg body weight for males and females, respectively. Prior to the animal study comprehensive analytical characterization of both rice materials was performed. The chemical analyses showed a number of statistically significant differences, with the majority being within the ranges reported in the literature. In the animal study a range of clinical, biological, immunological, microbiological and pathological parameters were examined. A number of significant differences were seen between groups fed the two diets, but none of them were considered to be adverse. In conclusion, the design of the present animal study did not enable us to conclude on the safety of the GM food. Additional group(s) where the expressed gene products have been spiked to the diet should be included in order to be able to distinguish whether the observed effects were due to the GNA lectin per se or to secondary changes in the GM rice. Publication Types: Evaluation Studies Research Support, Non-U.S. Gov't PMID: 17052828 [PubMed - indexed for MEDLINE] 608: Theriogenology. 2007 Jan 1;67(1):166-77. Epub 2006 Oct 18. Compositional analysis of dairy products derived from clones and cloned transgenic cattle. Laible G, Brophy B, Knighton D, Wells DN. AgResearch, Ruakura Research Centre, Hamilton, New Zealand. goetz.laible@agresearch.co.nz Cloning technology is an emerging biotechnological tool that could provide commercial opportunities for livestock agriculture. However, the process is very inefficient and the molecular events underlying the technology are poorly understood. The resulting uncertainties are causing concerns regarding the safety of food products derived from cloned livestock. There are similar concerns for livestock produced by biotechnologies which enable the purposeful introduction of genetic modifications. To increase the knowledge about food products from animals generated by these modern biotechnologies, we assessed compositional differences associated with milk and cheese derived from cloned and transgenic cows. Based on gross composition, fatty acid and amino acid profiles and mineral and vitamin contents, milk produced by clones and conventional cattle were essentially similar and consistent with reference values from dairy cows farmed in the same region under similar conditions. Whereas colostrum produced by transgenic cows with additional casein genes had similar IgG secretion levels and kinetics to control cows, milk from the transgenic cows had a distinct yellow appearance, in contrast to the white color of milk from control cows. Processing of milk into cheese resulted in differences in the gross composition and amino acid profiles; 'transgenic' cheese had lower fat and higher salt contents and small but characteristic differences in the amino acid profile compared to control cheese. Publication Types: Comparative Study Research Support, Non-U.S. Gov't PMID: 17052749 [PubMed - indexed for MEDLINE] 609: Food Chem Toxicol. 2007 Mar;45(3):339-49. Epub 2006 Sep 8. A 90-day safety study of genetically modified rice expressing Cry1Ab protein (Bacillus thuringiensis toxin) in Wistar rats. Schrøder M, Poulsen M, Wilcks A, Kroghsbo S, Miller A, Frenzel T, Danier J, Rychlik M, Emami K, Gatehouse A, Shu Q, Engel KH, Altosaar I, Knudsen I. Department of Toxicology and Risk Assessment, Danish Institute for Food and Veterinary Research, Mørkhøj Bygade 19, DK-2860 Søborg, Denmark. An animal model for safety assessment of genetically modified foods was tested as part of the SAFOTEST project. In a 90-day feeding study on Wistar rats, the transgenic KMD1 rice expressing Cry1Ab protein was compared to its non-transgenic parental wild type, Xiushui 11. The KMD1 rice contained 15mg Bt toxin/kg and based on the average feed consumption the daily intake was 0.54mg Bt toxin/kg body weight. No adverse effects on animal behaviour or weight gain were observed during the study. Blood samples collected one week prior to sacrifice were analyzed and compared for standard haematological and biochemical parameters. A few parameters were significantly different, but all within the normal reference intervals for rats of this breed and age and not in relation to any other findings, thus not considered treatment related. Upon sacrifice a large number of organs were weighed, macroscopic and histopathological examinations were performed with only minor changes to report. The aim of the study was to use a known animal model in performance of safety assessment of a GM crop, in this case KMD1 rice. The results show no adverse or toxic effects of KMD1 rice when tested in the design used in this 90-day study. Nevertheless the experiences from this study lead to the overall conclusion that safety assessment for unintended effects of a GM crop cannot be done without additional test group(s). Publication Types: Evaluation Studies Research Support, Non-U.S. Gov't PMID: 17050059 [PubMed - indexed for MEDLINE] 610: Neuroscience. 2007 Jan 5;144(1):17-25. Epub 2006 Oct 13. The contribution of endogenous opioids to food reward is dependent on sex and background strain. Hayward MD, Low MJ. Center for the Study of Weight Regulation, Oregon Health and Science University, 3181 Southwest Sam Jackson Park Road, Portland, OR 97239, USA. Michael.Hayward@Xenogen.com Complex behaviors such as those associated with reward to unconditioned positive reinforcers are polygenic processes. In studies using genetically modified mice specific for the endogenous opioid systems an observed phenotype in a complex behavior is likely to be dependent on interacting genes which, in inbred mouse lines, influence that phenotype. To address this issue we examined operant responding for palatable food reinforcers in mice lacking the expression of beta-endorphin, enkephalin or both peptides congenic to two different genetic backgrounds; C57BL/6J and DBA/2J. These two inbred strains were chosen because their endogenous opioid states differ and they respond differently to exogenous opioids in many behavioral assays. We found that wildtype and mutant C57BL/6J mice acquired operant responding for food reinforcers faster than DBA/2J mice, regardless of their opioid genotype. Although wildtype DBA/2J mice had a significant deficit in acquisition of bar-pressing behavior to reach a pre-established performance criterion, no subsequent deficit was observed under two different schedules of reinforcement. Additionally, we found that mice lacking enkephalin had decreased motivation to bar press for palatable food reinforcers under a progressive ratio regardless of sex or background strain. In contrast, the only subset of beta-endorphin-deficient mice that had decreased motivation to bar press under a progressive ratio was males on the C57BL/6J background. Of the two classical endogenous opioid peptides with preferential activation of the mu opioid receptor, the knockout models would suggest that enkephalins play a more consistent role than beta-endorphin in mediating the motivation for food reward when tested under a progressive ratio. Publication Types: Research Support, N.I.H., Extramural PMID: 17049174 [PubMed - indexed for MEDLINE] 611: Nutr Metab Cardiovasc Dis. 2007 Feb;17(2):74-81. Epub 2006 Oct 13. Assessment of usual dietary intake in population studies of gene-diet interaction. Tucker KL. Jean Mayer USDA Human Nutrition Research Center on Aging, Tufts University, 711 Washington Street, Boston, MA 02111, USA. katherine.tucker@tufts.edu AIMS: Dietary intake is a critical environmental exposure when considering the effect of many genetic factors on disease risk. However, dietary intake is a complex and changing measure that requires particular care in assessment. DATA SYNTHESIS: Although weighed diet records can theoretically provide the most accurate assessment of intake, they are usually not realistic in large population studies due to heavy respondent burden, likelihood of poor compliance, and the cost of data entry. Multiple 24-h dietary recalls can provide excellent detail, allowing for diverse dietary practices, but they are costly and require multiple contacts with participants. Food frequency questionnaires are the most cost-effective tool for assessing usual intake, particularly for micronutrients with high day-to-day variability. However, they have limitations for diverse populations and recent studies have questioned their ability to measure macronutrient intakes for assessing diet and disease relationships. CONCLUSION: At the present time, food frequencies remain the most cost-effective tool for large population studies. However, their limitations must be fully appreciated and demonstration of validity for nutrients of concern in the populations under study is essential. When macronutrients are of key interest, consideration should be given to the use of multiple recalls. Records may be used only in educated and compliant populations. Continued efforts to improve dietary assessment methodology must be investigated. Publication Types: Review PMID: 17046222 [PubMed - indexed for MEDLINE] 612: Nat Biotechnol. 2006 Oct;24(10):1178. Comment in: Nat Biotechnol. 2007 Jan;25(1):33-4. Parallel universes? [No authors listed] An EU Commissioner has a meeting of minds with an antibiotech agitator. Publication Types: Editorial PMID: 17033639 [PubMed - indexed for MEDLINE] 613: Nat Biotechnol. 2006 Oct;24(10):1177. Comment in: Nat Biotechnol. 2007 Feb;25(2):165; discussion 165-6. Why silence is not an option. [No authors listed] GM products will continue to be marginalized in Europe as long as industry remains silent. Publication Types: Editorial PMID: 17033637 [PubMed - indexed for MEDLINE] 614: Methods Mol Biol. 2006;344:135-41. Pine (Pinus radiata). Grant J, Dale T, Cooper P. New Zealand Institute for Crop and Food Research Ltd., Christchurch, New Zealand. This chapter describes the transformation of Pinus radiata using organogenic cotyledon explants rather than the more common somatic embryogenesis methods for conifers. The advantages of our method are the year round availability of seed and that over 80% of genotypes can be easily regenerated from the mature cotyledon explants. The transformation efficiency (i.e., the number of transformed shoots regenerated from excised cotyledons) is 1.7% and, as with other Agrobacterium tumefaciens transformation methods, the majority of transgene integrations are single copy. Critical factors for success are survival of the cotyledons, Agrobacterium strain, and selection pressure after cocultivation. PMID: 17033058 [PubMed - indexed for MEDLINE] 615: Methods Mol Biol. 2006;344:25-36. Potato (Solanum tuberosum L.). Millam S. Institute of Molecular Plant Science, University of Edinburgh, EH9 3JR, The United Kingdom. Potato (Solanum tuberosum L.) is a globally important crop plant producing high yields of nutritionally valuable food in the form of tubers. It has been the focus of substantial study because of its use both as a staple food crop and as a potentially significant source of compounds of interest. This has included the development and application of transgenic technology for introducing novel traits of fundamental and applied interest. This chapter describes a rapid, efficient, and cost-effective system for the routine transformation of this crop plant at rates above 40% efficiency, calculated as the mean number of Southern blot- confirmed independent transgenics per number of internodal explants originally plated. Internodal sections are co-cultivated with Agrobacterium tumefaciens and subjected to a two-stage callus induction/shoot outgrowth system under kanamycin selection. Shoot regeneration rates are high using the described method, and excised independent shoots rooting from the cut end of the stem after two further subcultures on kanamycin are 95% certain to be transformed. The transgenic status can be confirmed by molecular analysis and the plants grown on for tuber production enabling a wide spectrum of further studies. Publication Types: Research Support, Non-U.S. Gov't PMID: 17033048 [PubMed - indexed for MEDLINE] 616: Biotechnol Lett. 2006 Dec;28(23):1877-88. Epub 2006 Sep 22. IgE binding to proteins from sesame and assessment of allergenicity: implications for biotechnology? Orruño E, Morgan MR. Procter Department of Food Science, University of Leeds, Leeds, UK, prceo@leeds.ac.uk Successful prediction of the potential allergenicity of a protein may be a key factor in the development of novel, genetically modified foods. The use of the decision tree approach for the prediction of allergenicity is discussed. The methods currently used for identifying allergenic proteins (including use of IgE from patient sera for recognition of proteins) are reviewed. Finally, a specific review of the literature concerning identification of allergens from sesame leads to the conclusion that in the absence of validated animal models, identification of allergenicity (and, consequently, prediction of allergenicity) may be problematic. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 17028779 [PubMed - indexed for MEDLINE] 617: Regul Toxicol Pharmacol. 2007 Feb;47(1):37-47. Epub 2006 Oct 5. Compositional assessment of event DAS-59122-7 maize using substantial equivalence. Herman RA, Storer NP, Phillips AM, Prochaska LM, Windels P. Dow AgroSciences LLC, Indianapolis, IN 46268, USA. raherman@dow.com Event DAS-59122-7 (Herculex RW) maize (Zea mays L.) plants were transformed to express the Cry34Ab1 and Cry35Ab1 binary insecticidal crystal proteins originally isolated from Bacillus thuringiensis Berliner (Bt) strain PS149B1. These proteins protect maize roots from attack by corn rootworms, Diabrotica spp. DAS-59122-7 maize also contains the pat gene, originally isolated from Streptomyces viridochromogenes, which confers tolerance to glufosinate-ammonium herbicides (e.g. Liberty). We assessed the composition of these transgenic plants (with and without Liberty herbicide treatment), grown at a total of eight fields sites over 2 years, by applying the principle of substantial equivalence. Forage and grain samples were analyzed for proximates, fiber and minerals, and grain was further analyzed for amino acids, fatty acids, vitamins, secondary metabolites and anti-nutrients. Data plots were prepared that allow for efficient investigation of equivalency between event DAS-59122-7 maize and a non-transgenic near-isogenic maize line grown contemporaneously. Results demonstrated that DAS-59122-7 maize is equivalent to non-transgenic maize with respect to these important constituents. Publication Types: Comparative Study PMID: 17027131 [PubMed - indexed for MEDLINE] 618: Prikl Biokhim Mikrobiol. 2006 Jul-Aug;42(4):485-8. [Accuracy of a real-time polymerase-chain-reaction assay for a quantitative estimation of genetically modified sources in food products] [Article in Russian] Abramov DD, Trofimov DIu, Rebrikov DV. The accuracy of a real-time polymerase-chain-reaction assay for genetically modified sources in food products was determined using two official test systems (kits) of primers and samples. These kits were recommended by the Federal Center of State Sanitary and Epidemiological Surveillance (Russian Ministry of Health) and the European Commission. We used the following three models of thermocyclers: iCycler iQ (BioRad, United States), Rotor-Gene 3000 (Corbett Research, Australia), and DT-322 (DNA-Technology, Russia). Studies of samples that contained 1% genetically modified sources showed that the error of a quantitative assay for genetically modified sources in food products corresponds to 20-30% and does not depend on the kit type and the thermocycler model used. Publication Types: Comparative Study English Abstract PMID: 17022461 [PubMed - indexed for MEDLINE] 619: Environ Toxicol Chem. 2006 Oct;25(10):2653-61. Subacute effects of transgenic crylab bacillus thuringiensis corn litter on the isopods Trachelipus rathkii and armadillidium nasatum. Clark BW, Prihoda KR, Coats JR. Department of Entomology and Interdepartmental Toxicology Program, Iowa State University, 115 Insectary, Ames, Iowa 50011, USA. Laboratory studies were conducted to investigate the subacute effects of transgenic Cry1Ab corn leaf material containing Bacillus thuringiensis (Bt) protein on the terrestrial isopods Trachelipus rathkii and Armadillidium nasatum. Survival and growth were measured for eight weeks in isopods fed leaf material of two Bt11 corn varieties, two Monsanto 810 (Mon810) corn varieties, and the isolines of each. Total lipid and protein content of the organisms was measured to examine effects on energetic reserves. Armadillidium nasatum individuals in all treatments responded similarly. For T. rathkii, no statistically significant effect of Bt was observed, but statistical differences were observed in growth between hybrids. Protein and sugar content of the food were found to be correlated with the differences in growth for T. rathkii. Total protein content was higher in T. rathkii and A. nasatum fed material with higher protein and sugar content. A trend toward less growth in T. rathkii on Bt corn varieties versus their isolines triggered a concentration-response assay with purified Cry1Ab protein. No adverse effects of purified Bt protein were observed. These results indicate that little hazard to T. rathkii and A. nasatum from Bt corn leaf material from these hybrids exists. However, nutritional differences in corn hybrids contributed to differences in isopod growth. PMID: 17022406 [PubMed - indexed for MEDLINE] 620: J Vet Med Sci. 2006 Sep;68(9):959-65. Involvement of neuropeptide Y in hyperphagia in human growth hormone transgenic rats. Hozumi H, Yamanouchi K, Nishihara M. Department of Veterinary Physiology, Veterinary Medical Science, The University of Tokyo, Tokyo, Japan. We have previously produced human growth hormone (hGH) transgenic (TG) rats that show low circulating levels of both hGH and endogenous rat GH. Although body length of the TG rats is normal, they develop hyperphagia and severe obesity. The present study was undertaken to elucidate the causes of hyperphagia in the TG rats by focusing on temporal changes in plasma ghrelin levels and hypothalamic neuropeptide Y (NPY) contents. In both wild-type (WT) and TG rats, the highest value of plasma ghrelin levels was observed just before the dark phase, and thereafter plasma ghrelin levels were maintained higher in the TG than WT rats. Although NPY contents also showed the peak level just before the dark phase in both the arcuate (ARC) and paraventricular nuclei (PVN) of the hypothalamus, the values in the ARC, but not the PVN, of the TG rats was always lower than those of the WT rats, suggesting increased transport of NPY from the ARC to PVN in the TG rats. In addition, treatment with antagonists for Y1 and Y5 receptors for NPY reduced food intake much more effectively in the TG than WT rats. Intermittent treatment with recombinant hGH for a week significantly decreased food consumption, adipose tissue weight and plasma triglyceride concentrations in the TG rats. These results suggest that, in the TG rats, insufficiency in circulating GH stimulates the ghrelin-NPY system with a resultant increase in food intake. PMID: 17019066 [PubMed - indexed for MEDLINE] 621: Biotechnol Genet Eng Rev. 2004;21:325-67. Design of safe and biologically contained transgenic plants: tools and technologies for controlled transgene flow and expression. Gleba Y, Marillonnet S, Klimyuk V. Icon Genetics AG, Biozentrum Halle, Weinbergweg 22, D-06120 Halle/Saale, Germany. gleba@icongenetics.de Publication Types: Review PMID: 17017039 [PubMed - indexed for MEDLINE] 622: Biotechnol Genet Eng Rev. 2004;21:299-324. Safety testing and regulation of genetically engineered foods. Freese W, Schubert D. Friends of the Earth U.S., 1717 Massachusetts Ave., NW, Suite 600, Washington, DC 20036, USA. Publication Types: Review PMID: 17017038 [PubMed - indexed for MEDLINE] 623: Acta Trop. 2006 Oct;99(2-3):173-83. Epub 2006 Sep 26. Re-introducing bacteria in mosquitoes--a method for determination of mosquito feeding preferences based on coloured sugar solutions. Lindh JM, Terenius O, Eriksson-Gonzales K, Knols BG, Faye I. Department of Genetics, Microbiology and Toxicology, Stockholm University, 106 91 Stockholm, Sweden. In this study, sugar-feeding was investigated as a possible means of re-introducing bacteria into mosquito midguts with the aim of identifying bacteria that are suitable for creating paratransgenic mosquitoes. In a paratransgenic approach, bacteria are utilised to deliver effector molecules capable of inhibiting pathogen development in the midgut of the vector. To determine if mosquitoes discriminate between sterile sugar solutions and sugar solutions with bacteria, a method for screening mosquito feeding preferences was developed. This method was tested for Aedes aegypti, Anopheles arabiensis and An. gambiae s.s. mosquitoes and is based on a dual-choice test of solutions labelled with food dyes. Three different tests (dye/colour detection, sugar detection and sugar-concentration detection) were performed to evaluate the method, after which bacteria previously isolated from mosquitoes were used in the experiments. It was shown that mosquitoes do not discriminate between sugar solutions with or without these bacteria indicating that sugar-feeding is a possible means to introduce bacteria into mosquitoes. Furthermore, two different setups of the method were used, enabling us to differentiate between tactile/taste and olfactory responses. The method described in this paper is easy to use, cost-effective and allows broad screening of mosquito sugar-feeding preferences. Publication Types: Research Support, Non-U.S. Gov't PMID: 16999928 [PubMed - indexed for MEDLINE] 624: Methods Mol Biol. 2006;343:407-14. Tepary bean (Phaseolus acutifolius). Zambre M, Van Montagu M, Angenon G, Terryn N. Insitute Plant Biotechnology for Developing Countries, University of Ghent, Gent, Belgium. Phaseolus beans are among the major legumes for food consumption, especially in Latin America, Africa, and Asia. Tepary bean (Phaseolus acutifolius L. Gray) is one of the five cultivated species of the genus Phaseolus. This chapter describes an Agrobacterium-mediated transformation protocol for P. acutifolius based on cocultivation of callus, derived from cotyledonary nodes, with Agrobacterium. The selectable marker gene used is neomycin phosphotransferase II (nptII), and the selection agent is geneticin. Selection of transgenic callus material is achieved through four to five passages on geneticin-containing medium, after which shoots are induced on medium without selection agent. The protocol as described here has been applied to transform a cultivated variety of P. acutifolius, TB1, and also with some modifications to a wild genotype, NI576 and another cultivated variety, PI440795. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 16988363 [PubMed - indexed for MEDLINE] 625: Methods Mol Biol. 2006;343:359-67. Pigeonpea (Cajanus cajan L. Millsp.). Sharma KK, Sreelatha G, Dayal S. Genetic Transformation Laboratory, International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Andhra Pradesh, India. Pigeonpea [Cajanus cajan (L.) Millsp.], also known as redgram, is one of the major grain legume (pulses) crops grown in the semiarid tropics (SAT) extending between 30 degrees N and 30 degrees S; it is the second most important food legume of India. It is cultivated in about 50 countries of Asia, Africa, and the Americas for a variety of uses (food, fodder, fuel wood, rearing lac insects, hedges, wind breaks, soil conservation, green manure, roofing, and so on). The constraints of enhancing its productivity include the damage caused by various fungi, bacteria, viruses, and insect pests. Conventional plant breeding methods have not been successful for the improvement of pigeonpea because of genetic variation and incompatibility among the wild varieties. Genetic engineering technology can therefore be used as an additional tool for the introduction of agronomically useful traits into established varieties. The development of plant transformation techniques has been a major breakthrough in overcoming constraints to achieve precision in genetic manipulation. The development of efficient plant regeneration protocols is a prerequisite for recombinant technology to carry out genetic transformation. This chapter describes an Agrobacterium-mediated transformation protocol for pigeonpea, a simple, efficient, and reproducible method that is applicable across diverse genotypes of pigeonpea. Publication Types: Review PMID: 16988359 [PubMed - indexed for MEDLINE] 626: Methods Mol Biol. 2006;343:337-45. Peas (Pisum sativum L.). Grant J, Cooper P. New Zealand for Crop & Food Research, Ltd., Christchurch, New Zealand. In this chapter we describe a robust method for transformation of peas that has been successfully used in our laboratory since 1992. Using immature pea seed collected from field- or greenhouse-grown plants, we have produced transgenic lines for over 30 genotypes including named pea cultivars and advanced breeding lines. This method uses immature cotyledons as the explant, and the transformation efficiency is in the range 0.2 to 13.5% of cotyledons producing at least one independently transformed line. Agrobacterium tumefaciens strains AGL1 and KYRT1 are the most successful in our procedure, and kanamycin, phosphinothricin, and hygromycin are reliable selectable markers. Potentially useful genes have been introduced for pest and disease resistance, altering quality traits, and investigating metabolic pathways and are being studied in transgenic pea lines. Publication Types: Review PMID: 16988357 [PubMed - indexed for MEDLINE] 627: Methods Mol Biol. 2006;343:313-23. Chickpea (Cicer arietinum L.). Sharma KK, Bhatnagar-Mathur P, Jayanand B. Genetic Transformation Laboratory, International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Andhra Pradesh, India. Chickpea is one of the most important leguminous, cool-season, food crops, cultivated prevalently in the Asian Pacific region. In spite of its nutritional importance, its area of cultivation has been low, with virtually no increase. Conventional breeding has resulted in several important improvements in this crop, and recent advances in biotechnology such as plant tissue culture and genetic transformation can significantly contribute to better sustainability of this important food crop. Here, we describe an efficient Agrobacterium-mediated transformation protocol for chickpea using axillary meristem explants, which results in a high frequency of genetic transformation (70%) and recovery of valuable transgenic plants. The protocol is significant owing to its high reproducibility and recovery of the transgenics in a relatively short period (90-100 days). Publication Types: Review PMID: 16988355 [PubMed - indexed for MEDLINE] 628: Methods Mol Biol. 2006;343:201-12. Indica rice (Oryza sativa, BR29 and IR64). Datta K, Datta SK. International Rice Research Institute, Plant Breeding, Genetics, and Biotechnology Division, Tissue Culture and Genetic Engineering Laboratory, Metro Manila, Philippines. Rice is the world's most important food crop. Indica-type rice provides the staple food for more than half of the world population. To satisfy the growing demand of the ever-increasing population, more sustained production of indica-type rice is needed. In addition, because of the high per capita consumption of indica rice, improvement of any traits including its nutritive value may have a significant positive health outcome for the rice-consuming population. Rice yield productivity is greatly affected by different biotic stresses, like diseases and insect pests, and abiotic stresses like drought, cold, and salinity. Attempts to improve resistance in rice to these stresses by conventional breeding through introgression of traits have limited success owing to a lack of resistance germplasm in the wild relatives. Gene transfer technology with genes from other sources can be used to make rice plants resistant or tolerant to insect pests, diseases, and different environmental stresses. For improving the nutritional value of the edible endosperm part of the rice, genes for increasing iron, beta-carotene, or better quality protein can be introduced in rice plants by genetic engineering. Different crops have been transformed using various gene transfer methods, such as protoplast transformation, biolistic, and Agrobacterium-mediated transformation. This chapter describes the Agrobacterium-mediated transformation protocol for indica-type rice. The selectable marker genes used are hygromycin phosphotransferase (hpt), neomycin phosphotransferase (nptII), or phosphomannose isomerase (pmi), and, accordingly, the selection agents are hygromycin, kanamycin (G418), or mannose, respectively. Publication Types: Review PMID: 16988345 [PubMed - indexed for MEDLINE] 629: Wei Sheng Yan Jiu. 2006 Jul;35(4):431-4. [Stability of hpt marker gene in transgenic rice in different food matrices and under varying food-processing conditions] [Article in Chinese] Shen LM, Wu YN, Zhang JZ, Zhou PP. Institute of Nutrition and Food Safety, Chinese Center for Disease Control and Prevention, Beijing 100050, China. OBJECTIVE: To estimate the likelihood of horizontal gene transfer from transgenic rice to bacteria of the food chain and human gut, the stability of s86 transgenic rice hpt gene in different food matrices and under varying food-processing conditions was studied. METHODS: Degradation of DNA was monitored by fragment -multiplex polymerase chain reaction. Integrity of hpt gene in various food samples was tested. RESULTS: A PCR system for the hpt gene of genetically modified rice has been established to detect fragments ranging between 236bp and 910bp. Detection of hpt and rbcl gene fragments was carried out in various food-processed samples by this PCR system. The data showed that the fragments up to 500 bp were detected in rice and congee, while the fragment length more than 236bp was not detected in crispy rice and popcorn-like rice. CONCLUSION: These results suggested that there are significant differences in DNA degradation by different food-processing methods. The likelihood of the large hpt gene fragments transfer from transgenic rice processed food to bacteria is reduced by food process. Publication Types: English Abstract PMID: 16986517 [PubMed - in process] 630: Shokuhin Eiseigaku Zasshi. 2006 Aug;47(4):146-50. Detection method for genetically modified papaya using duplex PCR. Yamaguchi A, Shimizu K, Mishima T, Aoki N, Hattori H, Sato H, Ueda N, Watanabe T, Hino A, Akiyama H, Maitani T. Japan Food Research Laboratories, Chitose: 2-3, Bunkyo, Chitose-shi, Hokkaido 066-0052, Japan. A simple and rapid method for the identification of genetically modified (GM) papaya, derived from Line 55-1, was developed by modifying the Japanese official PCR method. Genomic DNA was directly extracted from the fresh fruit without the lyophilization step, using a commercial silica-based kit. To develop a duplex PCR method which simultaneously detects the GM papaya-specific gene and the intrinsic papain gene, the papain 2-5'/3' (amplicon size; 184 bp) primer pair for the detection of the papain gene was newly designed within the region of the products (211 bp) amplified using the papain 1-5'/-3' primer pair adopted in the Japanese official PCR method. To detect the GM papaya-specific gene, the primer pair Nos C-5'/CaM N-3' described in the Japanese official method was used. The DNA sequences of the GM papaya gene and the intrinsic papain gene were co-amplified using the PCR method in a single tube. The developed duplex PCR method allows the simultaneous detection of the products by means of agarose gel electrophoresis or microchip electrophoresis. The proposed method for GM papaya identification is simple and rapid. Publication Types: Research Support, Non-U.S. Gov't PMID: 16984033 [PubMed - indexed for MEDLINE] 631: Regul Toxicol Pharmacol. 2007 Feb;47(1):90-5. Epub 2006 Sep 18. ELISA method for monitoring human serum IgE specific for Cry1Ab introduced into genetically modified corn. Nakajima O, Teshima R, Takagi K, Okunuki H, Sawada J. National Institute of Health Sciences, Division of Biochemistry and Immunochemistry, 1-18-1 Kamiyoga, Setagaya, Tokyo, Japan. onakajim@nihs.go.jp Enzyme-linked immunosorbent assay (ELISA) is the most convenient method of monitoring the occurrence of IgE antibodies specific for novel proteins in genetically modified (GM) foods. The levels of IgE specific for a recombinant protein, Cry1Ab, were determined using an ELISA method. A soluble form of the Cry1Ab protein purified from pCold1 vector-transformed Escherichia coli pTf16/BL21 was used as the ELISA coating antigen, and 1M NaCl was used as the washing buffer to remove IgE non-specifically bound to the coated antigen. Sera from 44 patients allergic to major food allergens were obtained, diluted 20-fold, tested, and found no identifiable IgE above background levels. We also tested sera from patients with corn allergy against whole extracts of non-GM and GM-corn (MON 810) using immunoblotting. The staining patterns were similar for the two types of corn. These results indicate that significant levels of IgE antibodies specific to Cry1Ab were not found in the sera of Japanese patients with food allergies. Publication Types: Research Support, Non-U.S. Gov't PMID: 16982119 [PubMed - indexed for MEDLINE] 632: Nucleic Acids Res. 2006;34(13):3779-93. Epub 2006 Aug 23. Computational detection of allergenic proteins attains a new level of accuracy with in silico variable-length peptide extraction and machine learning. Soeria-Atmadja D, Lundell T, Gustafsson MG, Hammerling U. Division of Toxicology, National Food Administration, Uppsala, Sweden The placing of novel or new-in-the-context proteins on the market, appearing in genetically modified foods, certain bio-pharmaceuticals and some household products leads to human exposure to proteins that may elicit allergic responses. Accurate methods to detect allergens are therefore necessary to ensure consumer/patient safety. We demonstrate that it is possible to reach a new level of accuracy in computational detection of allergenic proteins by presenting a novel detector, Detection based on Filtered Length-adjusted Allergen Peptides (DFLAP). The DFLAP algorithm extracts variable length allergen sequence fragments and employs modern machine learning techniques in the form of a support vector machine. In particular, this new detector shows hitherto unmatched specificity when challenged to the Swiss-Prot repository without appreciable loss of sensitivity. DFLAP is also the first reported detector that successfully discriminates between allergens and non-allergens occurring in protein families known to hold both categories. Allergenicity assessment for specific protein sequences of interest using DFLAP is possible via ulfh@slv.se. Publication Types: Evaluation Studies Research Support, Non-U.S. Gov't PMID: 16977698 [PubMed - indexed for MEDLINE] 633: Electrophoresis. 2006 Oct;27(19):3879-88. A new PCR-CGE (size and color) method for simultaneous detection of genetically modified maize events. Nadal A, Coll A, La Paz JL, Esteve T, Pla M. Institut de Tecnologia Agroalimentària, Universitat de Girona, EPS, Girona, Spain. We present a novel multiplex PCR assay for simultaneous detection of multiple transgenic events in maize. Initially, five PCR primers pairs specific to events Bt11, GA21, MON810, and NK603, and Zea mays L. (alcohol dehydrogenase) were included. The event specificity was based on amplification of transgene/plant genome flanking regions, i.e., the same targets as for validated real-time PCR assays. These short and similarly sized amplicons were selected to achieve high and similar amplification efficiency for all targets; however, its unambiguous identification was a technical challenge. We achieved a clear distinction by a novel CGE approach that combined the identification by size and color (CGE-SC). In one single step, all five targets were amplified and specifically labeled with three different fluorescent dyes. The assay was specific and displayed an LOD of 0.1% of each genetically modified organism (GMO). Therefore, it was adequate to fulfill legal thresholds established, e.g., in the European Union. Our CGE-SC based strategy in combination with an adequate labeling design has the potential to simultaneously detect higher numbers of targets. As an example, we present the detection of up to eight targets in a single run. Multiplex PCR-CGE-SC only requires a conventional sequencer device and enables automation and high throughput. In addition, it proved to be transferable to a different laboratory. The number of authorized GMO events is rapidly growing; and the acreage of genetically modified (GM) varieties cultivated and commercialized worldwide is rapidly increasing. In this context, our multiplex PCR-CGE-SC can be suitable for screening GM contents in food. Publication Types: Research Support, Non-U.S. Gov't PMID: 16972302 [PubMed - indexed for MEDLINE] 634: J Neuroendocrinol. 2006 Oct;18(10):776-85. Exaggerated response of arginine vasopressin-enhanced green fluorescent protein fusion gene to salt loading without disturbance of body fluid homeostasis in rats. Fujio T, Fujihara H, Shibata M, Yamada S, Onaka T, Tanaka K, Morita H, Dayanithi G, Kawata M, Murphy D, Ueta Y. Department of Occupational Health, Matsushita Science Center of Industrial Hygiene, Kadoma, Japan. We examined the effects of chronic salt loading on the hypothalamic expressions of the enhanced green fluorescent protein (eGFP), arginine vasopressin (AVP) and oxytocin (OXT) genes in AVP-eGFP transgenic rats that expressed eGFP in the hypothalamic AVP-containing neurones. In these rats, salt loading for 5 days caused a marked increase of the eGFP fluorescence in the magnocellular divisions of the paraventricular nucleus (PVN), the supraoptic nucleus (SON) and the internal layer of the median eminence. Expression of the eGFP gene was increased seven- to eight-fold in the PVN and SON of salt-loaded rats in comparison with euhydrated rats. By contrast, none of these changes were observed in the suprachiasmatic nucleus. The expression of the AVP and OXT genes was increased 1.5- to two-fold in the PVN and SON of salt-loaded nontransgenic (control) and transgenic rats. There were no differences in the expression levels of the AVP and OXT genes in the PVN and SON between nontransgenic (control) and transgenic animals under normal conditions and after salt loading. In the posterior pituitary gland, the intensity of the eGFP fluorescence did not change after salt loading for 5 days, but increased after 10 days of salt loading. Upon salt loading, significant increases in the plasma AVP concentrations, plasma osmolality and plasma Na+ were observed. Furthermore, there were no significant differences in changes of water intake, food intake, urine volume, urine osmolality, urine Na+ concentrations, and the body weights in both models under normal or salt-loaded conditions. Our results show that the response of the AVP-eGFP fusion gene to chronic salt loading is exaggerated, and humoral responses such as AVP and OXT and the body fluid homeostasis are maintained in AVP-eGFP transgenic rats. The AVP-eGFP transgenic rat gives us a new opportunity to study the dynamics of the AVP system in vivo. Publication Types: Research Support, Non-U.S. Gov't PMID: 16965296 [PubMed - indexed for MEDLINE] 635: J Comp Physiol A Neuroethol Sens Neural Behav Physiol. 2006 Dec;192(12):1335-48. Epub 2006 Sep 9. Olfactory conditioning of proboscis activity in Drosophila melanogaster. Chabaud MA, Devaud JM, Pham-Delègue MH, Preat T, Kaiser L. Développement, Evolution et Plasticité du Système Nerveux, CNRS, Bât. 32/33, Avenue de la Terrasse, 91198, Gif-sur-Yvette cedex, France. Olfactory learning and memory processes in Drosophila have been well investigated with aversive conditioning, but appetitive conditioning has rarely been documented. Here, we report for the first time individual olfactory conditioning of proboscis activity in restrained Drosophila melanogaster. The protocol was adapted from those developed for proboscis extension conditioning in the honeybee Apis mellifera. After establishing a scale of small proboscis movements necessary to characterize responses to olfactory stimulation, we applied Pavlovian conditioning, with five trials consisting of paired presentation of a banana odour and a sucrose reward. Drosophila showed conditioned proboscis activity to the odour, with a twofold increase of percentage of responses after the first trial. No change occurred in flies experiencing unpaired presentations of the stimuli, confirming an associative basis for this form of olfactory learning. The adenylyl cyclase mutant rutabaga did not exhibit learning in this paradigm. This protocol generated at least a short-term memory of 15 min, but no significant associative memory was detected at 1 h. We also showed that learning performance was dependent on food motivation, by comparing flies subjected to different starvation regimes. Publication Types: Research Support, Non-U.S. Gov't PMID: 16964495 [PubMed - indexed for MEDLINE] 636: Nat Biotechnol. 2006 Sep;24(9):1075-7. Why spurning food biotech has become a liability. Miller HI, Conko G, Kershen DL. Hoover Institution, Stanford University, Stanford, California 94305-6010, USA. miller@hoover.stanford.edu PMID: 16964211 [PubMed - indexed for MEDLINE] 637: Methods Mol Biol. 2006;342:321-34. Transgene-like animal models using intronic microRNAs. Lin SL, Chang SJ, Ying SY. Department of Cell and Neurobiology, Keck School of Medicine, University of Southern California, Los Angeles, CA, USA. Transgenic animal models are valuable tools for testing gene functions and drug mechanisms in vivo. They are also the best similitude of a human body for etiological and pathological research of diseases. All pharmaceutically developed drugs must be proven safe and effective in animals before approval by the Food and Drug Administration to be used in clinical trials. To this end, the transgenic animal models of human diseases serve as a front line for drug evaluation. However, there is currently no transgenic animal model for microRNA (miRNA) research. miRNAs, small single-stranded regulatory RNAs capable of silencing intracellular gene transcripts that contain either complete or partial complementarity to the miRNAs, are useful for the design and development of new therapies against cancer polymorphism and viral mutation. Recently, varieties of natural miRNAs have been found to be derived from hairpin-like RNA precursors in almost all eukaryotes, including yeast (Schizosaccharomyces pombe), plant (Arabidopsis), nematode (Caenorhabditis elegans), fly (Drosophila melanogaster), fish, mouse, and human, involving intracellular defense against viral infections and regulation of certain gene expressions during development. To facilitate the miRNA research in vivo, we have developed a state-of-the-art transgenic strategy for silencing specific genes in zebrafish, chicken, and mouse, using intronic miRNAs. By insertion of a hairpin-like pre-miRNA structure into the intron region of a gene, we have found that mature miRNAs were successfully transcribed by RNA polymerase (Pol)-II, coexpressed with the encoding gene transcript, and excised out of the encoding gene transcript by natural RNA splicing and processing mechanisms. In conjunction with retroviral transfection systems, the hairpin-like pre-miRNA construct was further inserted into the intron of a cellular gene for tissue-specific expression regulated by the gene promoter. Because the retroviral vectors were randomly integrated into the genome of its host cell, the most effective transgenic animal can be selected and propagated to be a stable transgenic line for future research. Here, we have shown for the first time that transgene-like animal models were generated using the intronic miRNA-expressing system described previously, which has been proven to be useful for both miRNA research and in vivo evaluation of miRNA-associated target genes. Publication Types: Research Support, N.I.H., Extramural Review PMID: 16957386 [PubMed - indexed for MEDLINE] 638: Curr Biol. 2006 Aug 8;16(15):R563-4. GMOs still rankle in Europe. Williamson N. Publication Types: News PMID: 16953534 [PubMed - indexed for MEDLINE] 639: Carcinogenesis. 2007 Feb;28(2):471-8. Epub 2006 Aug 31. 4-monochlorobiphenyl (PCB3) induces mutations in the livers of transgenic Fisher 344 rats. Lehmann L, L Esch H, A Kirby P, W Robertson L, Ludewig G. Institute of Applied Biosciences, Section of Food Chemistry and Toxicology, University of Karlsruhe (TH) Kaiserstrasse 12, D-76131 Karlsruhe, Germany. 4-monochlorobiphenyl (PCB3) is found in small amounts in commercial PCB mixtures, indoor and outdoor air, and in food. In contrast to highly chlorinated congeners that are more resistant to metabolic attack, PCB3 is more readily converted by xenobiotic-metabolizing enzymes to monohydroxy-PCBs and further to dihydroxy-metabolites, which can be oxidized to quinones. Our recent studies demonstrated the initiating action of PCB3 in the livers of male rats. Therefore we hypothesized that PCB3 and/or its metabolite(s) are mutagenic in rat livers in vivo. To investigate the mutagenicity and the types of mutations generated by PCB3, male Fischer 344 BigBlue rats, transgenic for the lacI gene, were injected intraperitoneally with PCB3 (600 micromol/kg), 4-hydroxy-PCB3 (4-HO-PCB3, 400 micromol/kg), 3-methylcholanthrene (3-MC, 300 micromol/kg, positive control) and corn oil (negative control) once per week, for 4 weeks. Animals were killed 17 days after the last injection and the mutant frequency of the liver lacI gene determined. 3-MC induced a 4-fold increase of the mutant frequency of the lacI gene in the liver. The mutant frequency in PCB3-treated animals was also significantly elevated. In contrast, 4-HO-PCB3 induced a non-significant doubling of the mutant frequency. The mutation spectrum of solvent control mutants was characterized by transitions, whereas in 3-MC-animals, transversion and frameshift mutations predominated. The PCB3-induced mutation spectrum was similar to that of the 3-MC-induced mutants. In contrast, the mutation spectrum of the 4-HO-PCB3 group hardly differed from that of the control animals. This study demonstrates for the first time the mutagenicity of a PCB in vivo. Publication Types: Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. PMID: 16950798 [PubMed - indexed for MEDLINE] 640: J Allergy Clin Immunol. 2006 Sep;118(3):711-8. Epub 2006 Jul 12. Skin prick tests reveal stable and heritable reduction of allergenic potency of gene-silenced tomato fruits. Lorenz Y, Enrique E, Lequynh L, Fötisch K, Retzek M, Biemelt S, Sonnewald U, Vieths S, Scheurer S. Paul-Ehrlich-Institut, Langen, Germany. BACKGROUND: Today, for patients with food allergy, the only possibility to prevent allergic reactions is avoidance of the allergenic food. Genetic engineering of hypoallergenic plants by means of RNA interference (RNAi) could be an approach to improve the quality of life of subjects with food allergy. OBJECTIVES: We sought to achieve stable inhibition of expression of the allergenic nonspecific lipid transfer protein Lyc e 3 in tomato and to analyze the reduction of allergenicity in vitro by using histamine release assays and in vivo by using skin prick tests with transgenic tomato fruits. METHODS: Gene silencing was performed by means of RNAi and monitored by using Western blotting with nonspecific lipid transfer protein-specific antibodies and sera from patients with tomato allergy. Dose-dependent basophil histamine release assays, prick-to-prick skin testing, and determination of endogenous histamine content were performed with fruits harvested from plants of the first and second generation to assess the allergenic potency compared with that of wild-type fruits. RESULTS: We demonstrated that silencing of Lyc e 3 by means of RNAi contributes to reduced skin reactivity and is passed on to the next generation of fruits. A significant reduction of allergenic potency was determined in vitro and confirmed by using skin prick tests. CONCLUSION: Taken together, these results indicate that RNAi technology is an effective tool to generate foods with reduced allergenicity. CLINICAL IMPLICATIONS: Allergen-reduced plant foods might allow reduction of dietary restrictions for patients allergic to panallergen families. Publication Types: Research Support, Non-U.S. Gov't PMID: 16950292 [PubMed - indexed for MEDLINE] 641: BMC Genomics. 2006 Aug 31;7:222. Complete plastid genome sequence of Daucus carota: implications for biotechnology and phylogeny of angiosperms. Ruhlman T, Lee SB, Jansen RK, Hostetler JB, Tallon LJ, Town CD, Daniell H. Department of Molecular Biology & Microbiology, University of Central Florida, Biomolecular Science, Building #20, Room 336, Orlando, FL 32816-2364, USA. daniell@mail.ucf.edu BACKGROUND: Carrot (Daucus carota) is a major food crop in the US and worldwide. Its capacity for storage and its lifecycle as a biennial make it an attractive species for the introduction of foreign genes, especially for oral delivery of vaccines and other therapeutic proteins. Until recently efforts to express recombinant proteins in carrot have had limited success in terms of protein accumulation in the edible tap roots. Plastid genetic engineering offers the potential to overcome this limitation, as demonstrated by the accumulation of BADH in chromoplasts of carrot taproots to confer exceedingly high levels of salt resistance. The complete plastid genome of carrot provides essential information required for genetic engineering. Additionally, the sequence data add to the rapidly growing database of plastid genomes for assessing phylogenetic relationships among angiosperms. RESULTS: The complete carrot plastid genome is 155,911 bp in length, with 115 unique genes and 21 duplicated genes within the IR. There are four ribosomal RNAs, 30 distinct tRNA genes and 18 intron-containing genes. Repeat analysis reveals 12 direct and 2 inverted repeats > or = 30 bp with a sequence identity > or = 90%. Phylogenetic analysis of nucleotide sequences for 61 protein-coding genes using both maximum parsimony (MP) and maximum likelihood (ML) were performed for 29 angiosperms. Phylogenies from both methods provide strong support for the monophyly of several major angiosperm clades, including monocots, eudicots, rosids, asterids, eurosids II, euasterids I, and euasterids II. CONCLUSION: The carrot plastid genome contains a number of dispersed direct and inverted repeats scattered throughout coding and non-coding regions. This is the first sequenced plastid genome of the family Apiaceae and only the second published genome sequence of the species-rich euasterid II clade. Both MP and ML trees provide very strong support (100% bootstrap) for the sister relationship of Daucus with Panax in the euasterid II clade. These results provide the best taxon sampling of complete chloroplast genomes and the strongest support yet for the sister relationship of Caryophyllales to the asterids. The availability of the complete plastid genome sequence should facilitate improved transformation efficiency and foreign gene expression in carrot through utilization of endogenous flanking sequences and regulatory elements. Publication Types: Comparative Study Research Support, N.I.H., Extramural Research Support, U.S. Gov't, Non-P.H.S. PMID: 16945140 [PubMed - indexed for MEDLINE] 642: Yonsei Med J. 2006 Aug 31;47(4):505-12. Evaluating the allergic risk of genetically modified soybean. Kim SH, Kim HM, Ye YM, Kim SH, Nahm DH, Park HS, Ryu SR, Lee BO. Department of Allergy, Ajou University School of Medicnie, Suwon, Korea. hspark@ajou.ac.kr Genetically modified (GM) soybean (carrying the EPSPS transgene) is the most common GM food in Korea. In order to assess whether genetic modification increases the allergenic risk of soybeans, the allergenicity and IgE-reactive components of wild-type and GM soybean extracts were compared in allergic adults who had been sensitized to soybeans. We enrolled 1,716 adult allergy patients and 40 healthy, non-atopic controls. Skin prick tests and IgE enzyme linked immunosorbent assays (ELISAs) were performed using wild-type and GM soybean extracts, along with other common inhaled allergens. The specificities of serum IgE antibodies from allergic patients and the identities of the IgE-reactive components of the soybean extracts were compared using ELISA inhibition testing, 2-dimensional gel electrophoresis, and IgE immunoblotting. To evaluate the effects of digestive enzymes and heat treatment, the soybean extracts were heated or pre- incubated with or without simulated gastric and intestinal fluids. The IgE sensitization rates to wild-type and GM soybeans were identical (3.8% of allergic adults), and circulating IgE antibodies specific for the two extracts were comparable. The results of the ELISA inhibition test, SDS-PAGE, and IgE immunoblotting showed a similar composition of IgE-binding components within the wild-type and GM extracts, which was confirmed using two-dimensional gel electrophoresis, IgE immunoblotting, and amino acid sequencing. None of the subjects had a positive response to purified EPSPS protein in the skin prick test, ELISA, or IgE immunoblot analysis. These findings suggest that the IgE sensitization rate to GM soybean extracts is identical to that of wild-type soybean extracts in adult allergy patients. In addition, based on both in vivo and in vitro methods, the allergenicity of wild type and GM soybean extracts was identical. Publication Types: Research Support, Non-U.S. Gov't PMID: 16941740 [PubMed - indexed for MEDLINE] 643: J Econ Entomol. 2006 Aug;99(4):1085-95. Effect of corn hybrids expressing the coleopteran-specific cry3Bb1 protein for corn rootworm control on aboveground insect predators. Ahmad A, Wilde GE, Whitworth RJ, Zolnerowich G. Department of Entomology, Kansas State University, Manhattan, KS 66506-4004, USA. aahmad@ksu.edu Field and laboratory studies were conducted to determine the effect of transgenic Bacillus thuringiensis (Bt) corn, Zea mays L. (YieldGard Rootworm), expressing the Cry3Bb1 protein on aboveground nontarget insect predators (minute pirate bug, ladybird beetles, and carabids). Visual counts of adult and immature Orius insidiosus (Say), Coleomegilla maculata (DeGeer), Hippodamia convergens Gurin-Meneville, and Scymnus spp. occurring in Bt corn and its non-Bt isoline were made at Manhattan, KS, in 2002 and at Manhattan and Scandia, KS, in 2003. No significant differences were found between the Bt corn and non-Bt isoline plots in the abundance (number per plant) of O. insidiosus, C. maculata, H. convergens, and Scymnus spp. Field predation on Ostrinia nubilalis (Hübner) (Lepidoptera: Crambidae) egg masses was also observed during the silking stage of corn at Manhattan and Scandia in 2003. No significant differences were observed among treatments in predation rate for predators with chewing versus sucking mouthparts. Two laboratory studies determined the effect of Cry3Bb1 protein expressed in Bt corn pollen on C. maculata and carabids. The larvae of C. maculata were reared on Bt pollen, non-Bt pollen, or greenbugs, Schizaphis graminum (Rondani). The duration of larval and pupal stages, developmental time from egg hatch to adult emergence, percentage of survival, and elytra length were compared among treatments. There were no significant differences in developmental time of larvae fed pollen or greenbugs during their first two instars. However, significantly prolonged development of the third (1 d) and fourth instars (2 d) was observed for larvae fed greenbugs only. Total time for larval development was significantly longer for larvae that fed on greenbugs versus larvae fed on pollen. No significant differences were observed among treatments in the percentage of larvae that pupated or pupal stage duration. Larvae that fed on greenbugs had higher pupal and adult weights compared with pollen-fed larvae. However, pupal and adult weights did not vary between the Bt and non-Bt pollen treatments. No significant differences occurred in longevity and elytra length of beetles among all treatments. Two carabid species, Harpalus caliginosus F. and Harpalus pensylvanicus DeGeer, were reared on moistened dog food sprinkled with Bt or non-Bt corn pollen. No significant differences in mortality of H. caliginosus and H. pensylvanicus were detected among any of the treatments. There was no significant effect of Bt pollen on fecundity and egg viability of H. caliginosus. Our studies showed that YieldGard Rootworm had no effect on the selected coleopteran predators; therefore, this Bt corn hybrid could be used in an integrated pest management system. PMID: 16937659 [PubMed - indexed for MEDLINE] 644: Plasmid. 2007 Jan;57(1):18-28. Epub 2006 Aug 28. A role for the tet(O) plasmid in maintaining Campylobacter plasticity. Friis LM, Pin C, Taylor DE, Pearson BM, Wells JM. Institute of Food Research, Colney Lane, Norwich, Norfolk NR4 7UA, UK. lfriis@ualberta.ca Genomic sequencing projects are beginning to reveal regions of extensive DNA homology between bacterial genera. Public fears of the spread of genetically modified organisms into the food chain and the increasing prevalence of multi-drug resistant disease in humans highlight the implications of horizontal gene transfer. The striking DNA sequence similarity between the two uniquely identified tetracycline resistant (Tc(R)) Campylobacter plasmids, pCC31 and pTet, suggests their conserved acquisition and maintenance within Campylobacter [Batchelor, R.A., Pearson, B.M., Friis, L.M., Guerry, P., Wells, J.M. 2004. Nucleotide sequences and comparison of two large conjugative plasmids from different Campylobacter species. Microbiology 150, 3507-3517]. It is thus likely that these and other conjugative plasmids are highly prevalent and broadly distributed across several continents. Microarray technology is now enabling fast and extensive genomic comparisons to be made and allows us to investigate intra- and inter-genetic conservation and variability. This study details the development of a microarray specific for genes from Campylobacter plasmids pCC31, pTet and pVir and its application to the analysis of Campylobacter plasmid gene presence and preservation throughout environmental and clinical isolates. Application of the iterative algorithm GENCOM (freely available at ) is used as a rapid and effective way of comparing the content and conservation of plasmids in bacteria and provides details of the Campylobacter flexible gene pool and its contribution to genomic plasticity. Publication Types: Research Support, Non-U.S. Gov't PMID: 16934869 [PubMed - indexed for MEDLINE] 645: Plant Biol (Stuttg). 2006 Sep;8(5):662-72. Epub 2006 Aug 24. Evaluation of a non-targeted "omic" approach in the safety assessment of genetically modified plants. Metzdorff SB, Kok EJ, Knuthsen P, Pedersen J. Danish Institute for Food and Veterinary Research, 19 Mørkhøj Bygade, 2860 Søborg, Denmark. metz@dfvf.dk Genetically modified plants must be approved before release in the European Union, and the approval is generally based upon a comparison of various characteristics between the transgenic plant and a conventional counterpart. As a case study, focusing on safety assessment of genetically modified plants, we here report the development and characterisation of six independently transformed ARABIDOPSIS THALIANA lines modified in the flavonoid biosynthesis. Analyses of integration events and comparative analysis for characterisation of the intended effects were performed by PCR, quantitative Real-time PCR, and High Performance Liquid Chromatography. Analysis by cDNA microarray was used as a non-targeted approach for the identification of potential unintended effects caused by the transformation. The results revealed that, although the transgenic lines possessed different types of integration events, no unintended effects were identified. However, we found that the majority of genes showing differential expression were identified as stress-related genes and that environmental conditions had a large impact on the expression of several genes, proteins, and metabolites. We suggest that the microarray approach has the potential to become a useful tool for screening of unintended effects, but state that it is crucial to have substantial information on the natural variation in traditional crops in order to be able to interpret "omics" data correctly within the framework of food safety assessment strategies of novel plant varieties, including genetically modified plant varieties. Publication Types: Comparative Study Research Support, Non-U.S. Gov't PMID: 16933176 [PubMed - indexed for MEDLINE] 646: Mol Cell Proteomics. 2006 Dec;5(12):2228-43. Epub 2006 Aug 22. Proteomics fingerprinting of phagosome maturation and evidence for the role of a Galpha during uptake. Gotthardt D, Blancheteau V, Bosserhoff A, Ruppert T, Delorenzi M, Soldati T. Department of Molecular Cell Research, Max Planck Institute for Medical Research, University Hospital of Heidelberg, Germany. Phagocytosis, whether of food particles in protozoa or bacteria and cell remnants in the metazoan immune system, is a conserved process. The particles are taken up into phagosomes, which then undergo complex remodeling of their components, called maturation. By using two-dimensional gel electrophoresis and mass spectrometry combined with genomic data, we identified 179 phagosomal proteins in the amoeba Dictyostelium, including components of signal transduction, membrane traffic, and the cytoskeleton. By carrying out this proteomics analysis over the course of maturation, we obtained time profiles for 1,388 spots and thus generated a dynamic record of phagosomal protein composition. Clustering of the time profiles revealed five clusters and 24 functional groups that were mapped onto a flow chart of maturation. Two heterotrimeric G protein subunits, Galpha4 and Gbeta, appeared at the earliest times. We showed that mutations in the genes encoding these two proteins produce a phagocytic uptake defect in Dictyostelium. This analysis of phagosome protein dynamics provides a reference point for future genetic and functional investigations. Publication Types: Research Support, Non-U.S. Gov't Validation Studies PMID: 16926386 [PubMed - indexed for MEDLINE] 647: Sci Am. 2006 Sep;295(3):24. Old MacDonald's pharm. Choi CQ. Publication Types: News PMID: 16925026 [PubMed - indexed for MEDLINE] 648: Crit Rev Biotechnol. 2006 Jul-Sep;26(3):121-43. Biotechnological methods to accelerate cheddar cheese ripening. Azarnia S, Robert N, Lee B. Department of Food Science and Agricultural Chemistry, McGill University, Ste-Anne-de-Bellevue, QC, Canada. Cheese is one of the dairy products that can result from the enzymatic coagulation of milk. The basic steps of the transformation of milk into cheese are coagulation, draining, and ripening. Ripening is the complex process required for the development of a cheese's flavor, texture and aroma. Proteolysis, lipolysis and glycolysis are the three main biochemical reactions that are responsible for the basic changes during the maturation period. As ripening is a relatively expensive process for the cheese industry, reducing maturation time without destroying the quality of the ripened cheese has economic and technological benefits. Elevated ripening temperatures, addition of enzymes, addition of cheese slurry, attenuated starters, adjunct cultures, genetically engineered starters and recombinant enzymes and microencapsulation of ripening enzymes are traditional and modern methods used to accelerate cheese ripening. In this context, an up to date review of Cheddar cheese ripening is presented. Publication Types: Review PMID: 16923531 [PubMed - indexed for MEDLINE] 649: Public Health Nutr. 2006 Aug;9(5):662-3. Comment on: Public Health Nutr. 2005 Sep;8(6A):673-94. How far should nutrition reach? Kent G. Publication Types: Comment Letter PMID: 16923303 [PubMed - indexed for MEDLINE] 650: J Nutr. 2006 Sep;136(9):2331-7. Transgenic flavonoid tomato intake reduces C-reactive protein in human C-reactive protein transgenic mice more than wild-type tomato. Rein D, Schijlen E, Kooistra T, Herbers K, Verschuren L, Hall R, Sonnewald U, Bovy A, Kleemann R. Friedrich-Alexander-Universität Erlangen-Nürnberg, D-91058 Erlangen, Germany. The increased consumption of fruits and vegetables is associated with reduced cardiovascular disease. The molecular basis of this health effect is not fully understood, yet dietary flavonoids are thought to play an important role. Genetic engineering has enabled us to overexpress specific flavonoids (flavones and flavonols) in tomato fruit. Human C-reactive protein transgenic (CRPtg) mice express markers of cardiovascular risk that allow us to study of the putative health effects of wild-type tomato (wtTom) and flavonoid-enriched tomato (flTom). In this study, we analyzed whether consumption of wtTom, at a dose achievable with a human diet, has beneficial effects on cardiovascular risk markers and whether flTom may enhance such effects. CRPtg mice were fed a diet containing 4 g/kg wtTom, flTom peel, vehicle, or 1 g/kg fenofibrate, which reportedly reduces cardiovascular risk, for 7 wk. Markers of general health (bodyweight, food intake, and plasma alanine aminotransferase activities) and of cardiovascular risk (plasma CRP, fibrinogen, E-selectin, and cholesterol levels) were analyzed. All groups had comparable food intakes and body-weight gains. Plasma alanine aminotransferase activities increased significantly in vehicle and fenofibrate-treated mice. Compared with baseline, wtTom and flTom significantly reduced basal human CRP concentrations by 43 and 56%, respectively. The CRP-lowering effect of flTom significantly exceeded that of wtTom. The effects of flTom on CRP were reversed within a 2-wk washout period. WtTom and flTom did not affect fibrinogen, but comparably repressed E-selectin expression and upregulated HDL cholesterol. Tomato peel consumption improved cardiovascular risk factors in CRPtg mice, a beneficial effect that was further enhanced by enrichment of the flavonoid content. Publication Types: Comparative Study Research Support, Non-U.S. Gov't PMID: 16920850 [PubMed - indexed for MEDLINE] 651: Food Chem Toxicol. 2006 Nov;44(11):1909-15. Epub 2006 Jul 4. Investigation on possible allergenicity of 19 different commercial enzymes used in the food industry. Bindslev-Jensen C, Skov PS, Roggen EL, Hvass P, Brinch DS. Department of Dermatology and Allergy Center, Odense University Hospital, DK 5000 Odense C, Denmark. Carsten.Bindslev-Jensen@ouh.fyns-amt.dk The aim of the study was to investigate the safety to allergic patients of 19 commercially available and authority-approved enzymes used in the food industry. Enzymes produced by genetically modified organisms were included. Four hundred consecutive adult patients with a diagnosed allergy to inhalation allergens, food allergens, bee or wasp were included. All had at least one positive skin prick test to the above allergens. Skin prick testing with the 19 enzymes was performed on the forearm and if positive (in 13 patients), in vitro histamine release from blood basophils were performed. Patients with positive results in skin prick test were subsequently reinvestigated with further purified enzymes and finally challenged orally with the enzymes in a double-blind, placebo-controlled protocol. Only one reaction to a placebo challenge was seen. In some instances a positive skin prick test result or a positive histamine release was seen elicited by the enzymes, but since none of the patients were positive to any of the commercial enzymes in the subsequent oral challenges using exaggerated dosages of the enzymes compared to normal daily intake, the findings are without clinical relevance. A wide variety of enzyme classes and origins was included in the study. Because there were no allergenic findings of clinical relevance it is concluded that ingestion of food enzymes in general is not considered to be a concern with regard to food allergy. Publication Types: Clinical Trial Controlled Clinical Trial PMID: 16920243 [PubMed - indexed for MEDLINE] 652: J AOAC Int. 2006 Jul-Aug;89(4):913-28. Immunoassay as an analytical tool in agricultural biotechnology. Grothaus GD, Bandla M, Currier T, Giroux R, Jenkins GR, Lipp M, Shan G, Stave JW, Pantella V. EnviroLogix Inc, 500 Riverside Industrial Pkwy, Portland, ME 04103, USA. davidgrothaus@envirologix.com Immunoassays for biotechnology engineered proteins are used by AgBiotech companies at numerous points in product development and by feed and food suppliers for compliance and contractual purposes. Although AgBiotech companies use the technology during product development and seed production, other stakeholders from the food and feed supply chains, such as commodity, food, and feed companies, as well as third-party diagnostic testing companies, also rely on immunoassays for a number of purposes. The primary use of immunoassays is to verify the presence or absence of genetically modified (GM) material in a product or to quantify the amount of GM material present in a product. This article describes the fundamental elements of GM analysis using immunoassays and especially its application to the testing of grains. The 2 most commonly used formats are lateral flow devices (LFD) and plate-based enzyme-linked immunosorbent assays (ELISA). The main applications of both formats are discussed in general, and the benefits and drawbacks are discussed in detail. The document highlights the many areas to which attention must be paid in order to produce reliable test results. These include sample preparation, method validation, choice of appropriate reference materials, and biological and instrumental sources of error. The article also discusses issues related to the analysis of different matrixes and the effects they may have on the accuracy of the immunoassays. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 16915826 [PubMed - indexed for MEDLINE] 653: Plant Mol Biol. 2006 Sep;62(1-2):71-82. Epub 2006 Aug 16. Self-excision of the antibiotic resistance gene nptII using a heat inducible Cre-loxP system from transgenic potato. Cuellar W, Gaudin A, Solórzano D, Casas A, Nopo L, Chudalayandi P, Medrano G, Kreuze J, Ghislain M. Applied Biotechnology Laboratory, Germplasm enhancement and Crop Improvement Division, International Potato Center CIP, P.O. Box 1558, Lima 12, Peru. Resistance to antibiotics mediated by selectable marker genes remains a powerful selection tool for transgenic event production. However, regulatory agencies and consumer concerns favor these to be eliminated from food crops. Several excision systems exist but none have been optimized or shown to be functional for clonally propagated crops. The excision of the nptII gene conferring resistance to kanamycin has been achieved here using a gene construct based on a heat-inducible cre gene producing a recombinase that eliminates cre and nptII genes flanked by two loxP sites. First-generation regenerants with the Cre-loxP system were obtained by selection on kanamycin media. Following a heat treatment, second generation regenerants were screened for excision by PCR using nptII, cre, and T-DNA borders primers. Excision efficiency appeared to be at 4.7% depending on the heat treatment. The footprint of the excision was shown by sequencing between T-DNA borders to correspond to a perfect recombination event. Selectable marker-free sprouts were also obtained from tubers of transgenic events when submitted to similar heat treatment at 4% frequency. Spontaneous excision was not observed out of 196 regenerants from untreated transgenic explants. Biosafety concerns are minimized because the expression of cre gene driven by the hsp70 promoter of Drosophila melanogaster was remarkably low even under heat activation and no functional loxP site were found in published Solanum sequence database. A new plant transformation vector pCIP54/55 was developed including a multiple cloning site and the self-excision system which should be a useful tool not only for marker genes in potato but for any gene or sequence removal in any plant. Publication Types: Research Support, Non-U.S. Gov't PMID: 16912912 [PubMed - indexed for MEDLINE] 654: Eur Rev Med Pharmacol Sci. 2006 Jul-Aug;10(4):197-206. Benefits and concerns associated with biotechnology-derived foods: can additional research reduce children health risks? Cantani A. Allergy and Clinical Immunology Division, Pediatric Department, La Sapienza University, Rome, Italy. The development of techniques devised for the genetic manipulation of foods poses new risks for children with food allergy (FA). The introduction of foreign allergenic proteins from different foods into previously tolerated foods may trigger allergic reactions, often complicating with anaphylactic shock in a subset of allergic babies. Children with FA, even if subjected to preventative diets, always challenge the risk of developing allergic manifestations after unintentional intake of a non tolerated food in restaurant settings, with relatives or schoolmates, etc, where product labelling is necessarily lacking. The introduction of potentially allergenic proteins into foods generally considered safe for allergic children can be done deliberately, by either substantially altering the food ingredients, or by genetic manipulation which change the composition or transfer allergens, or unintentionally by quality-control failures, due to contaminations in the production process, or to genetic mismanipulation. There is a controversy between multinationals often favored by governments and consumer association resistance, thus an equidistant analysis poses some unprecedented impediments. The importance of FA and the potential of transgenic plants to bring food allergens into the food supply should not be disregarded. The expression in soybeans of a Brazil nut protein resulted in a food allergen expressed in widely used infant formulas, so paving the way to an often reported multinational debacle. Genetic engineering poses innovative ethical and social concerns, as well as serious challenges to the environment, human health, animal welfare, and the future of agriculture. In this paper will be emphasized practical concepts more crucial for pediatricians. Publication Types: Review PMID: 16910351 [PubMed - indexed for MEDLINE] 655: J Environ Health. 2006 Jul-Aug;69(1):33-4. The ethical dilemma of genetically modified food. Jefferson V. National Capital Area Environmental Health Association, Clinton, MD 20735, USA. Val.Jefferson@verizon.net PMID: 16910106 [PubMed - indexed for MEDLINE] 656: BMC Biotechnol. 2006 Aug 14;6:37. Critical points of DNA quantification by real-time PCR--effects of DNA extraction method and sample matrix on quantification of genetically modified organisms. Cankar K, Stebih D, Dreo T, Zel J, Gruden K. Department of Plant Physiology and Biotechnology, National Institute of Biology, Vecna pot 111, 1000 Ljubljana, Slovenia. katja.cankar@nib.si BACKGROUND: Real-time PCR is the technique of choice for nucleic acid quantification. In the field of detection of genetically modified organisms (GMOs) quantification of biotech products may be required to fulfil legislative requirements. However, successful quantification depends crucially on the quality of the sample DNA analyzed. Methods for GMO detection are generally validated on certified reference materials that are in the form of powdered grain material, while detection in routine laboratories must be performed on a wide variety of sample matrixes. Due to food processing, the DNA in sample matrixes can be present in low amounts and also degraded. In addition, molecules of plant origin or from other sources that affect PCR amplification of samples will influence the reliability of the quantification. Further, the wide variety of sample matrixes presents a challenge for detection laboratories. The extraction method must ensure high yield and quality of the DNA obtained and must be carefully selected, since even components of DNA extraction solutions can influence PCR reactions. GMO quantification is based on a standard curve, therefore similarity of PCR efficiency for the sample and standard reference material is a prerequisite for exact quantification. Little information on the performance of real-time PCR on samples of different matrixes is available. RESULTS: Five commonly used DNA extraction techniques were compared and their suitability for quantitative analysis was assessed. The effect of sample matrix on nucleic acid quantification was assessed by comparing 4 maize and 4 soybean matrixes. In addition 205 maize and soybean samples from routine analysis were analyzed for PCR efficiency to assess variability of PCR performance within each sample matrix. Together with the amount of DNA needed for reliable quantification, PCR efficiency is the crucial parameter determining the reliability of quantitative results, therefore it was chosen as the primary criterion by which to evaluate the quality and performance on different matrixes and extraction techniques. The effect of PCR efficiency on the resulting GMO content is demonstrated. CONCLUSION: The crucial influence of extraction technique and sample matrix properties on the results of GMO quantification is demonstrated. Appropriate extraction techniques for each matrix need to be determined to achieve accurate DNA quantification. Nevertheless, as it is shown that in the area of food and feed testing matrix with certain specificities is impossible to define strict quality controls need to be introduced to monitor PCR. The results of our study are also applicable to other fields of quantitative testing by real-time PCR. Publication Types: Evaluation Studies Research Support, Non-U.S. Gov't PMID: 16907967 [PubMed - indexed for MEDLINE] 657: Pest Manag Sci. 2006 Oct;62(10):999-1012. Comparing the impact of conventional pesticide and use of a transgenic pest-resistant crop on the beneficial carabid beetle Pterostichus melanarius. Mulligan EA, Ferry N, Jouanin L, Walters KF, Port GR, Gatehouse AM. Institute for Research on Environment and Sustainability, School of Biology, University of Newcastle Upon-Tyne, Newcastle NE1 7RU, UK. The potential impact of a chemical pesticide control method has been compared with that of transgenic plants expressing a protease inhibitor conferring insect resistance by utilising a tritrophic system comprising the crop plant Brassica napus (L.) (Oilseed rape), the pest mollusc Deroceras reticulatum (Müller) and the predatory carabid beetle Pterostichus melanarius (Illiger). Cypermethrin, as the most widely used pesticide in UK oilseed rape (OSR) cultivation, was selected as the conventional treatment. OSR expressing a cysteine protease inhibitor, oryzacystatin-1 (OC-1), was the transgenic comparator. In feeding trials, D. reticulatum showed no significant long-term effects on measured life history parameters (survival, weight gain, food consumption) as a result of exposure to either the cypermethrin or OC-1 treatment. However, D. reticulatum was able to respond to the presence of the dietary inhibitor by producing two novel proteases following exposure to OC-1-expressing OSR. Similarly, P. melanarius showed no detectable alterations in mortality, weight gain or food consumption when feeding on D. reticulatum previously fed either pesticide-contaminated or GM plant material. Furthermore, as with the slug, a novel form of protease, approximately M(r) 27 kDa, was induced in the carabid in response to feeding on slugs fed OC-1-expressing OSR. Publication Types: Comparative Study Research Support, Non-U.S. Gov't PMID: 16906504 [PubMed - indexed for MEDLINE] 658: Transgenic Res. 2006 Aug;15(4):515-9. Consumption of milk from transgenic goats expressing human lysozyme in the mammary gland results in the modulation of intestinal microflora. Maga EA, Walker RL, Anderson GB, Murray JD. Department of Animal Science, University of California, One Shields Avenue, Davis, CA 95616, USA. eamaga@ucdavis.edu Lysozyme is a key antimicrobial component of human milk that has several health-promoting functions including the development of a healthy intestinal tract. However, levels of lysozyme in the milk of dairy animals are negligible. We have generated transgenic dairy goats that express human lysozyme (HLZ) in their milk in an attempt to deliver the benefits of human milk in a continual fashion. To test the feasibility of this transgenic approach to achieve a biological impact at the level of the intestine, feeding trials were conducted in two animal models. Pasteurized milk from HLZ transgenic animals was fed to both kid goats (ruminant model) and young pigs (human model), and the numbers of total coliforms and Escherichia coli present in the small intestine were determined. Data from this proof-of-principle study demonstrate that milk from transgenic animals was capable of modulating the bacterial population of the gut in both animal models. Pigs that consumed pasteurized milk from HLZ transgenic goats had fewer numbers of coliforms and E. coli in their intestine than did those receiving milk from non-transgenic control animals. The opposite effect was seen in goats. Milk from these transgenic animals not only represent one of the first transgenic food products with the potential of benefiting human health, but are also a unique model to study the development and role of intestinal microflora on health, well-being and resistance to disease. PMID: 16906451 [PubMed - indexed for MEDLINE] 659: Transgenic Res. 2006 Aug;15(4):465-80. Erratum in: Transgenic Res. 2007 Apr;16(2):253-9. Characterization and multi-generational stability of the growth hormone transgene (EO-1alpha) responsible for enhanced growth rates in Atlantic Salmon. Yaskowiak ES, Shears MA, Agarwal-Mawal A, Fletcher GL. Aqua Bounty Canada, Inc., P.O. Box 13422, A1B 4B7, St. John's, Newfoundland, Canada. Transgenic technologies provide a promising means by which desirable traits can be introduced into cultured fish species within a single generation thus accelerating the production of genetically superior broodstock for aquaculture. However, before such fish are allowed to be marketed as food they must receive government regulatory approval. Two pivotal regulatory requirements are: (1) complete characterization of the genomically integrated transgene and, (2) demonstration that the transgene remains stable over multiple generations. We have generated a stable line of growth hormone (GH) transgenic Atlantic salmon (Salmo salar) using an "all fish" gene construct (opAFP-GHc2) containing a growth hormone cDNA from chinook salmon whose expression is regulated by the 5' promoter and 3' termination regions derived from an ocean pout antifreeze protein (AFP) gene. In this study we show that a reorganized form of the opAFP-GHc2 construct (termed EO-1alpha) integrated as a single functional copy into a 35 bp repeat region of the genomic DNA. PCR based mapping revealed that the linear sequence of the EO-1alpha integrant was organized as follows: base pairs 1580-2193 of the ocean pout promoter region followed by the intact chinook salmon GH cDNA, the complete ocean pout antifreeze 3' region, and the first 1678 bp of the ocean pout antifreeze 5' region. Sequence analysis of the EO-1alpha integrant and genomic flanking regions in F2 and F4 generation salmon revealed that they were identical. In addition, apart from the disruption at the integration sites, the consensus sequences of the integrant in these two generations of salmon were identical to the sequence of the opAFP-GHc2 construct. These results indicate that the EO-1alpha transgene codes for the chinook salmon GH, and that the transgene and the integration site have remained stable over multiple generations. Publication Types: Research Support, Non-U.S. Gov't PMID: 16906447 [PubMed - indexed for MEDLINE] 660: Transgenic Res. 2006 Aug;15(4):409-25. Assessing the potential for unintended effects in genetically modified potatoes perturbed in metabolic and developmental processes. Targeted analysis of key nutrients and anti-nutrients. Shepherd LV, McNicol JW, Razzo R, Taylor MA, Davies HV. Quality, Health and Nutrition Programme, Scottish Crop Research Institute, Invergowrie, DD2 5DA, Dundee, Scotland. louise.shepherd@scri.ac.uk Targeted compositional analysis was carried out on transgenic potato tubers of either cultivar (cv.) Record or cv. Desirée to assess the potential for unintended effects caused by the genetic modification process. The range of transgenic lines analysed included those modified in primary carbohydrate metabolism, polyamine biosynthesis and glycoprotein processing. Controls included wildtype tubers, tubers produced from plants regenerated through tissue culture (including a callus phase) and tubers derived from transformation with the 'empty vector' i.e. no specific target gene included (with the exception of the kanamycin resistance gene as a selectable marker). Metabolite analysis included soluble carbohydrates, glycoalkaloids, vitamin C, total nitrogen and fatty acids. Trypsin inhibitor activity was also assayed. These cover the major compounds recommended by the OECD in their Consensus Document on Compositional Considerations for New Varieties of Potatoes: Key Food and Feed Nutrients, Anti-Nutrients and Toxicants (2002). Data was statistically analysed using analysis of variance (ANOVA) for individual compounds and, where applicable, principal component analysis (PCA). In general, targeted compositional analysis revealed no consistent differences between GM lines and respective controls. No construct specifically induced unintended effects. Statistically significant differences between wildtype controls and specific GM lines did occur but appeared to be random and not associated with any specific construct. Indeed such significant differences were also found between wildtypes and both tissue culture derived tubers and tubers derived from transformation with the empty vector. This raises the possibility that somaclonal variation (known to occur significantly in potato, depending on genotype) may be responsible for an unknown proportion of any differences observed between specific GM lines and the wildtype. The most obvious differences seen in GC-MS profiles were between the two potato varieties used in the study. Publication Types: Research Support, Non-U.S. Gov't PMID: 16906442 [PubMed - indexed for MEDLINE] 661: Transgenic Res. 2006 Aug;15(4):405-7. Cloned transgenic heart-healthy pork? Prather RS. Division of Animal Sciences, University of Missouri-Columbia, 920 East Campus Drive, E125 Animal Science Research centre, Columbia, MO 65211, USA. pratherr@missouri.edu Here I comment on the production and uses of swine that express a humanized fat-1 gene. The gene product is a fatty acid desaturase that converts omega-6 fatty acids to omega-3 fatty acids. Omega-3 fatty acids have been implicated as being important for reproductive success, maintaining a healthy cardiovascular system, sustaining a functional immune system, and even preventing depression and cancer. The descendants of these hfat-1 transgenic swine will be very useful as models of the human condition, and if they are permitted to enter the food chain, they may improve human health. Publication Types: Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't Review PMID: 16906441 [PubMed - indexed for MEDLINE] 662: Trends Genet. 2006 Oct;22(10):525-8. Epub 2006 Aug 9. Shall I compare thee to a GM potato? Colquhoun IJ, Le Gall G, Elliott KA, Mellon FA, Michael AJ. Institute of Food Research, Norwich Research Park, Colney, Norwich NR4 7UA, UK. A fundamental issue in the safety assessment of genetically modified crops is the question of whether unintentional changes have occurred in the crop plant as a consequence of the genetic modification. This question was addressed recently by using a powerful metabolite fingerprinting and metabolite profiling method to assess whether genetically modified potatoes are substantially similar to their corresponding conventional cultivars. Publication Types: Comparative Study PMID: 16904227 [PubMed - indexed for MEDLINE] 663: Food Drug Law J. 2006;61(2):197-235. Health and food safety: the benefits of Bt-corn. Kershen DL. University of Oklahoma College of Law, Norman, Okla., USA. PMID: 16903029 [PubMed - indexed for MEDLINE] 664: Food Drug Law J. 2006;61(2):167-96. The international regulation of genetically modified organisms: importing caution into the U.S. food supply. Strauss DM. Fairfield University, Charles F. Dolan School of Business, Fairfield, Conn., USA. PMID: 16903028 [PubMed - indexed for MEDLINE] 665: Nestle Nutr Workshop Ser Pediatr Program. 2006;58:207-15; discussion 215-7. Recombinant human milk proteins. Lönnerdal B. Department of Nutrition, University of California, Davis, CA, USA. Human milk provides proteins that benefit newborn infants. They not only provide amino acids, but also facilitate the absorption of nutrients, stimulate growth and development of the intestine, modulate immune function, and aid in the digestion of other nutrients. Breastfed infants have a lower prevalence of infections than formula-fed infants. Since many women in industrialized countries choose not to breastfeed, and an increasing proportion of women in developing countries are advised not to breastfeed because of the risk of HIV transmission, incorporation of recombinant human milk proteins into infant foods is likely to be beneficial. We are expressing human milk proteins known to have anti-infective activity in rice. Since rice is a normal constituent of the diet of infants and children, limited purification of the proteins is required. Lactoferrin has antimicrobial and iron-binding activities. Lysozyme is an enzyme that is bactericidal and also acts synergistically with lactoferrin. These recombinant proteins have biological activities identical to their native counterparts. They are equally resistant to heat processing, which is necessary for food applications, and to acid and proteolytic enzymes which are needed to maintain their biological activity in the gastrointestinal tract of infants. These recombinant human milk proteins may be incorporated into infant formulas, baby foods and complementary foods, and used with the goal to reduce infectious diseases. Publication Types: Review PMID: 16902336 [PubMed - indexed for MEDLINE] 666: Food Addit Contam. 2006 Sep;23(9):876-82. Development of an innovative immunoassay for CP4EPSPS and Cry1AB genetically modified protein detection and quantification. Ermolli M, Prospero A, Balla B, Querci M, Mazzeo A, Van Den Eede G. Biotechnology & GMOs Unit, Institute for Health and Consumer Protection (IHCP), European Commission, DG-Joint Research Centre (JRC), Via. E. Fermi 1, 201020 Ispra (VA), Italy. monica.ermolli@jrc.it An innovative immunoassay, called enzyme-linked immunoabsorbant assay (ELISA) Reverse, based on a new conformation of the solid phase, was developed. The solid support was expressly designed to be immersed directly in liquid samples to detect the presence of protein targets. Its application is proposed in those cases where a large number of samples have to be screened simultaneously or when the simultaneous detection of different proteins is required. As a first application, a quantitative immunoassay for Cry1AB protein in genetically modified maize was optimized. The method was tested using genetically modified organism concentrations from 0.1 to 2.0%. The limit of detection and limit of quantitation of the method were determined as 0.0056 and 0.0168 (expressed as the percentage of genetically modified organisms content), respectively. A qualitative multiplex assay to assess the presence of two genetically modified proteins simultaneously was also established for the case of the Cry1AB and the CP4EPSPS (5-enolpyruvylshikimate-3-phosphate synthase) present in genetically modified maize and soy, respectively. PMID: 16901856 [PubMed - indexed for MEDLINE] 667: Plant Foods Hum Nutr. 2006 Sep;61(3):145-50. Effect of cultivar, year grown, and cropping system on the content of tocopherols and tocotrienols in grains of hulled and hulless barley. Ehrenbergerová J, Belcrediová N, Prýma J, Vaculová K, Newman CW. Mendel University of Agriculture and Forestry, Brno, Czech Republic. ehren@mendelu.cz In a three-year period (2000-2002) total tocols (tocopherols and tocotrienols), content of vitamin E and its isomers (alpha-, beta+gamma-, delta-tocopherols and tocotrienols) were assessed in grain of 13 barley genotypes. The highest content of tocols (60.3-67.6 mg kg(-1)) and content of vitamin E (Vitamin E equivalent-18.0-20.1 mg kg(-1)) were determined in the waxy varieties Wanubet, Wabet, and Washonubet. Standard varieties, i.e. of a malting type (Krona and Kompakt), had statistically significantly lower content of tocols (49.9 and 53.6 mg kg(-1)) and vitamin E (15.7-16.1 mg kg(-1)) compared to the waxy varieties. The hulless waxy variety Washonubet had statistically significantly higher total content of tocols (67.6 mg kg(-1)) and alpha- tocotrienols isomer (42.1 mg kg(-1)) versus all other genotypes in the set. Chemical treatment and fertilization statistically significantly increased the content of tocols (by 4.7 mg kg(-1)), vitamin E (by 1.9 mg kg(-1)), isomer alpha-tocopherol (by 0.9 mg kg(-1)) and isomer alpha- tocotrienols (by 3.3 mg kg(-1)). The average values of alpha-tocopherols and alpha-tocotrienols in the set were 6.7 mg kg(-1) and 29.7 mg kg(-1), respectively. Some of the reciprocal lines created by us from the malting and waxy varieties are suitable for food use for high contents of all tocopherols and alpha-tocotrienols. Publication Types: Comparative Study Research Support, Non-U.S. Gov't PMID: 16900405 [PubMed - indexed for MEDLINE] 668: J Environ Qual. 2006 Aug 9;35(5):1633-58. Print 2006 Sep-Oct. The current status and environmental impacts of glyphosate-resistant crops: a review. Cerdeira AL, Duke SO. Brazilian Department of Agriculture, Agricultural Research Service, Embrapa/Environment, C.P. 69, Jaguariuna-SP-13820-000, Brazil. Glyphosate [N-(phosphonomethyl) glycine]-resistant crops (GRCs), canola (Brassica napus L.), cotton (Gossypium hirsutum L.), maize (Zea mays L.), and soybean [Glycine max (L.) Merr.] have been commercialized and grown extensively in the Western Hemisphere and, to a lesser extent, elsewhere. Glyphosate-resistant cotton and soybean have become dominant in those countries where their planting is permitted. Effects of glyphosate on contamination of soil, water, and air are minimal, compared to some of the herbicides that they replace. No risks have been found with food or feed safety or nutritional value in products from currently available GRCs. Glyphosate-resistant crops have promoted the adoption of reduced- or no-tillage agriculture in the USA and Argentina, providing a substantial environmental benefit. Weed species in GRC fields have shifted to those that can more successfully withstand glyphosate and to those that avoid the time of its application. Three weed species have evolved resistance to glyphosate in GRCs. Glyphosate-resistant crops have greater potential to become problems as volunteer crops than do conventional crops. Glyphosate resistance transgenes have been found in fields of canola that are supposed to be non-transgenic. Under some circumstances, the largest risk of GRCs may be transgene flow (introgression) from GRCs to related species that might become problems in natural ecosystems. Glyphosate resistance transgenes themselves are highly unlikely to be a risk in wild plant populations, but when linked to transgenes that may impart fitness benefits outside of agriculture (e.g., insect resistance), natural ecosystems could be affected. The development and use of failsafe introgression barriers in crops with such linked genes is needed. Publication Types: Review PMID: 16899736 [PubMed - indexed for MEDLINE] 669: Wiad Parazytol. 2003;49(1):11-20. [Transgenic bioinsecticides inimical to parasites, but imical to environment] [Article in Polish] Kucińska J, Lonc E, Rydzanicz K. Zakład Parazytologii Ogólnej, Instytut Mikrobiologii, Uniwersytet Wrocławski. Identification of Bacillus thuringiensis (Bt) parasporal crystalline inclusions composed of Cry proteins (=delta-endotoxins) resulted in introduction of microbial pesticides for biological control of some parasites. Delta-endotoxins are encoded by cry genes and are active against pest and nuisance insects (mostly mosquitoes and black flies--vectors of still important infectious diseases). The recent significant progress in DNA recombination technique may overcome limitations (a short residual persistence and a narrow spectrum of activity) associated with application of Bt conventional products. An introduction of cry genes from mosquitocidal subspecies B. th. israelensis (Bti) to the aquatic microorganisms inhabiting the same water bodies as mosquito and fly larvae (Diptera), has considerably improved the toxin delivery system to target insects. However, in the first experiments, in which Bti genes were cloned in cyanobacteria (Agmenellum quadruplicatum, Synechocystis PCC6803), a low gene expression was observed. Thus, it was necessary to integrate cry genes with strong promoters or to increase the number of vector-introduced copies. To overcome the obstacles of low gene expression and regulatory restriction for recombinant organisms, Bti spore/crystal formulations were encapsulated in the aquatic protozoan, Tetrahymena pyriformis. Large numbers of crystals (180 to 240/cell) were accumulated in its food vacuoles. This system resulted also in an increase in toxin persistence from 24 to 71 h. Cloning Bti genes in B. sphaericus (which also produces mosquitocidal proteins) was another way of an increasing Bt crystal residual activity. In this case, the crystals were additionally protected by B. sphaericus exosporium. These transgenic bacteria produced large amounts of delta-endotoxins that remained under water surface longer than the wild B. sphaericus strains. Moreover, they had a broader spectrum of insecticidal activity, because B. sphaericus is toxic mostly to Culex and Anopheles, and Bti--mostly to Culex, Aedes and some Simmulidae. Gram-negative bacteria (Asticcacaulis excentricus, Caulobacter crescentus and Ancylobacter aquaticus) turned out also to be effective delta-endotoxin producers. They grow on simple media and do not contain proteases which could degrade Cry proteins. In some cases, 100% mosquito larvae mortality was observed as a result of an exposure to transgenic microorganisms containing Bti genes. However, transgenic techniques are still not very popular in the world, despite their efficacy in biological control of insects. The transgenic organism construction is expensive and time-consuming. Genetic engineering is still raising a lot of anxieties and doubts concerning inappropriate use of modified organisms. On the other hand, this technology could solve many problems associated with vectors of important diseases, which are still unapproachable to contemporary medicine. Publication Types: English Abstract Review PMID: 16889013 [PubMed - indexed for MEDLINE] 670: Med Princ Pract. 2006;15(5):325-37. Human obesity: its hormonal basis and the role of gastric inhibitory polypeptide. Marks V. Department of Clinical Biochemistry, Post-Graduate Medical School, University of Surrey Guildford, Surrey, UK. vincentmarks@bigfoot.com Obesity is an abnormal expansion of the adipose organ and is a pathophysiological response to an imbalance between energy intake and energy expenditure. It is the result of a large number of diverse factors involving heritable and environmental characteristics. A simple definition of obesity is difficult and unsatisfactory and its age dependency has largely been ignored. Differentiation between healthy, age-related plumpness and obesity is often blurred and responsible for overdiagnosis of obesity in the developed world. In the past, epidemiological studies have often ignored the different prognostic significance of the two major phenotypes of human obesity making their conclusions of limited value. The role of heritable factors in determining both the propensity to develop obesity under favourable environmental conditions, including inactivity and unlimited access to fat-rich foods, and the phenotype it assumes received an enormous fillip from experiments involving genetically modified animals. The most important of these have demonstrated the key role played by a number of newly discovered or recently resurrected polypeptide hormones that are released from the intestine in response to food. Molecular manipulation of these hormones, especially of glucose-dependent insulin-stimulatory polypeptide offers a new therapeutic approach. Publication Types: Review PMID: 16888389 [PubMed - indexed for MEDLINE] 671: Environ Health Perspect. 2006 Aug;114(8):1154-7. Digestion assays in allergenicity assessment of transgenic proteins. Herman RA, Storer NP, Gao Y. Dow AgroSciences LLC, Indianapolis, Indiana 46268, USA. raherman@dow.com The food-allergy risk assessment for transgenic proteins expressed in crops is currently based on a weight-of-evidence approach that holistically considers multiple lines of evidence. This approach recognizes that no single test or property is known to distinguish allergens from nonallergens. The stability of a protein to digestion, as predicted by an in vitro simulated gastric fluid assay, currently is used as one element in the risk assessment process. A review of the literature on the use of the simulated gastric fluid assay to predict the allergenic status of proteins suggests that more extensive kinetic studies with well-characterized reference proteins are required before the predictive value of this assay can be adequately judged. Publication Types: Review PMID: 16882518 [PubMed - indexed for MEDLINE] 672: EMBO Rep. 2006 Aug;7(8):750-3. Cisgenic plants are similar to traditionally bred plants: international regulations for genetically modified organisms should be altered to exempt cisgenesis. Schouten HJ, Krens FA, Jacobsen E. Plant Research International, Wageningen University and Research Centre in the Netherlands. henk.schouten@wur.nl Publication Types: Research Support, Non-U.S. Gov't PMID: 16880817 [PubMed - indexed for MEDLINE] 673: Reprod Domest Anim. 2006 Aug;41(4):260-7. Erratum in: Reprod Domest Anim. 2006 Oct;41(5):477. Biosecurity and the various types of embryos transferred. Thibier M. Ministère de l'Agriculture et de la Pêche, Paris, France. michel.thibier@agriculture.gouv.fr The aim of the present paper was to review some features related to the risk analysis of three types of embryos to be transferred, namely the in vivo derived, the in vitro produced and the cloned ones. For in vivo-collected embryos, a considerable number of experiments and scientific investigations have been performed and hundreds of thousands of embryos are transferred annually with no contamination of associated diseases. Provided that the code of practice such as that published by the International Embryo Transfer Society is strictly followed by the embryo transfer practitioners, the statement made some 17 years ago saying that the in vivo-derived embryo transfer was the safest way of exchanging genes remains entirely true, thanks to the professionalism of the embryo transfer industry. For the in vitro-produced embryos, some particular rules have to be followed because of specific risks for some pathogens to strongly adhere to the zona pellucida of such embryos. There are some means to monitor and control those effects, and the transfer of in vitro-produced embryos can also be a very safe way to exchange genes around the world. The third type of embryos, the cloned ones, is a quite different category and the risk analysis to be soundly made still needs a lot of investigations so as to characterize the potential risks if there are, in terms not only of disease transmission but also in terms of public health, zoonotic risks as well as those related to quality and safety of food. The problem in this regard, is more directly addressed for offspring of clones than to the cloned embryos themselves. Published data on this issue are increasing in numbers so that progress in that area is expected in the few years to come. Publication Types: Review PMID: 16869879 [PubMed - indexed for MEDLINE] 674: Postepy Biochem. 2006;52(1):7-9. [Biochemistry as a background of modern biotechnology] [Article in Polish] Twardowski T. Instytut Chemii Bioorganicznej PAN i Politechnika Lódzka, ul. Noskowskiego 12/14, 61-704, Poznań. twardows@ibch.poznan.pl PMID: 16869296 [PubMed - indexed for MEDLINE] 675: Soc Reprod Fertil Suppl. 2006;62:303-15. Factors influencing the commercialisation of cloning in the pork industry. Pratt SL, Sherrer ES, Reeves DE, Stice SL. Department of Animal and Dairy Science, 425 River Road, Edgar Rhodes Animal and Dairy Science Complex, Athens, GA 30602, USA. scottprattc@AOL.com Production of cloned pigs using somatic cell nuclear transfer (SCNT) is a repeatable and predictable procedure and multiple labs around the world have generated cloned pigs and genetically modified cloned pigs. Due to the integrated nature of the pork production industry, pork producers are the most likely to benefit and are in the best position to introduce cloning in to production systems. Cloning can be used to amplify superior genetics or be used in conjunction with genetic modifications to produce animals with superior economic traits. Though unproven, cloning could add value by reducing pig-to-pig variability in economically significant traits such as growth rate, feed efficiency, and carcass characteristics. However, cloning efficiencies using SCNT are low, but predictable. The inefficiencies are due to the intrusive nature of the procedure, the quality of oocytes and/or the somatic cells used in the procedure, the quality of the nuclear transfer embryos transferred into recipients, pregnancy rates of the recipients, and neonatal survival of the clones. Furthermore, in commercial animal agriculture, clones produced must be able to grow and thrive under normal management conditions, which include attainment of puberty and subsequent capability to reproduce. To integrate SCNT into the pork industry, inefficiencies at each step of the procedure must be overcome. In addition, it is likely that non-surgical embryo transfer will be required to deliver cloned embryos, and/or additional methods to generate high health clones will need to be developed. This review will focus on the state-of-the-art for SCNT in pigs and the steps required for practical implementation of pig cloning in animal agriculture. Publication Types: Review PMID: 16866326 [PubMed - indexed for MEDLINE] 676: Shokuhin Eiseigaku Zasshi. 2006 Jun;47(3):111-4. A detection method of CryIAc protein for identifying genetically modified rice using the lateral flow strip assay. Akiyama H, Watanabe T, Kikuchi H, Sakata K, Tokishita S, Hayashi Y, Hino A, Teshima R, Sawada J, Maitani T. National Institute of Health Sciences: 1-18-1, Kamiyoga, Setagaya-ku, Tokyo 158-8501, Japan. We examined the lateral flow strip assay for identifying unauthorized genetically modified (GM) rice. The GM rice expresses the Bacillus thuringiensis (Bt) toxin, CryIAc protein, which confers tolerance to insects. The recombinant CryIAc protein was prepared from the inclusion bodies of an E. coli. strain into which the CryIAc gene had been inserted, using gel filtration chromatography. The lateral flow strip assay for the identification of GM cotton which also expresses CryIAc protein, was applied to unpolished rice and polished rice spiked with recombinant CryIAc protein. The spiked recombinant CryIAc protein was clearly detected at the level of 0.012 microg/g in both the unpolished and polished rice. After loading of the extract on the strip, a 60 -minute stand time is necessary to clearly detect CryIAc protein. The detection limit was approximately 12 ng CryIAc protein per gram of rice. These results suggest that the lateral flow strip assay for GM cotton can be used to detect CryIAc protein expressed in GM rice. Publication Types: Research Support, Non-U.S. Gov't PMID: 16862988 [PubMed - indexed for MEDLINE] 677: Obesity (Silver Spring). 2006 Jun;14(6):1003-9. Increased leptin expression in the dorsal vagal complex suppresses adiposity without affecting energy intake and metabolic hormones. Boghossian S, Lecklin A, Dube MG, Kalra PS, Kalra SP. Department of Neuroscience, University of Florida McKnight Brain Institute, Gainesville, 32610-0244, USA. OBJECTIVE: Increased leptin transgene expression locally in hypothalamic sites suppresses weight and energy intake, enhances thermogenic energy expenditure, and differentially modulates metabolic hormones for an extended period. We evaluated whether a similar localized expression of leptin transgene in the dorsal vagal complex (DVC) in the caudal brain stem that also displays the biologically relevant leptin receptor would reproduce these varied responses and thus demonstrate functional connectivity between the hypothalamus and DVC. RESEARCH METHODS AND PROCEDURES: Adult female rats were microinjected with a recombinant adeno-associated virus encoding either rat leptin or green fluorescent protein gene (control) in the DVC. Food intake and body weight were monitored weekly, and metabolic variables were analyzed at the end of 10 weeks. RESULTS AND DISCUSSION: Increased leptin transgene expression in the DVC suppressed the time-related increase in body weight accompanied by a transient decrease in food intake at week 1 post-injection and little effect on thermogenic energy expenditure. That suppression of weight was due to decreased adiposity is shown by the markedly suppressed white adipose tissue-derived hormones, leptin and adiponectin. Circulating concentrations of pancreatic insulin, gastric ghrelin, and glucose levels were unchanged. This segregation of the varied effects of leptin expression in hypothalamic sites vs. DVC endorses the view that among the various endocrine organs under sympathetic nervous system control, only those leptin-activated neural circuits in the hypothalamus that suppress weight and adiposity on a long-term basis transverse through DVC en route to white adipose tissue. Publication Types: Research Support, N.I.H., Extramural PMID: 16861605 [PubMed - indexed for MEDLINE] 678: J Biotechnol. 2006 Dec 15;127(1):161-6. Epub 2006 Jun 12. Random amplified polymorphic DNA analysis of genetically modified organisms. Yoke-Kqueen C, Radu S. Department of Biomedical Science, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia. ykcheah@medic.upm.edu.my Randomly amplified polymorphic DNA (RAPD) was used to analyzed 78 samples comprises of certified reference materials (soya and maize powder), raw seeds (soybean and maize), processed food and animal feed. Combination assay of two arbitrary primers in the RAPD analysis enable to distinguish genetically modified organism (GMO) reference materials from the samples tested. Dendrogram analysis revealed 13 clusters at 45% similarity from the RAPD. RAPD analysis showed that the maize and soybean samples were clustered differently besides the GMO and non-GMO products. PMID: 16860900 [PubMed - indexed for MEDLINE] 679: Genetics. 2006 Sep;174(1):309-16. Epub 2006 Jul 18. Effects of sex and insulin/insulin-like growth factor-1 signaling on performance in an associative learning paradigm in Caenorhabditis elegans. Vellai T, McCulloch D, Gems D, Kovács AL. Department of Genetics, Eötvös Loránd University, Budapest, H-1117, Hungary. vellai@falco.elte.hu Learning is an adaptive change in behavior in response to environmental stimuli. In mammals, there is a distinct female bias to learn skills that is still unprecedented in other animal taxa. Here we have investigated the biological determinants of performance in an associative learning paradigm in the nematode Caenorhabditis elegans. Using an assay of chemotactic reactions associated with food deprivation, wild-type male worms show inferior learning ability relative to hermaphrodites. Sex-based learning difference is therefore an ancient evolutionary feature appearing even in relatively simple animals. C. elegans mutants with reduced insulin/IGF-1 signaling also exhibit a greatly reduced learning ability in this assay. In addition, hyperactivation of insulin/IGF-1 signaling through loss-of-function mutations in the PTEN phosphatase daf-18, a negative regulator of insulin/IGF-1 signaling, enhances learning ability beyond that of wild type. According to our epistasis analysis, the effect of DAF-2 on learning acts via phosphatidylinositol 3,4,5-trisphosphate (PIP(3)) production, but not the DAF-16 FOXO transcription factor. This implies that the signaling pathway from DAF-2 affecting this learning paradigm branches between PIP(3) production and DAF-16. However, learning capacity of nematodes is lowered by loss-of-function mutations in daf-16, suggesting involvement of noninsulin/IGF-1 signaling-dependent DAF-16 activation in learning. Potentially, sex and insulin/IGF-1 signaling affect performance in this learning assay via effects on the neurobiology of learning. Publication Types: Comparative Study Research Support, Non-U.S. Gov't PMID: 16849598 [PubMed - indexed for MEDLINE] 680: Annu Rev Nutr. 2006;26:75-103. Innovative dietary sources of n-3 fatty acids. Whelan J, Rust C. Department of Nutrition, The University of Tennessee, Knoxville, Tennessee 37996-1920, USA. jwhelan@utk.edu It is now established that dietary n-3 polyunsaturated fatty acids (PUFAs) are involved in health promotion and disease prevention, particularly those traditionally derived from marine sources (e.g., eicosapentaenoic acid and docosahexaenoic acid). A number of organizations have made specific recommendations for the general population to increase their intakes of these nutrients. In response to and along with these recommendations, n-3 PUFAs are being incorporated into nontraditional food sources because of advances in the technology to safely enrich/fortify our food supply. Fatty acid compositions of traditional oils (e.g., canola and soybean) are being genetically modified to deliver more highly concentrated sources of n-3 PUFA. The advent of algal sources of docosahexaenoic acid provides one of the few terrestrial sources of this fatty acid in a concentrated form. All of this is possible because of newer technologies (microencapsulation) and improved processing techniques that ensure stability and preserve the integrity of these unstable fatty acids. Publication Types: Review PMID: 16848701 [PubMed - indexed for MEDLINE] 681: Planta. 2006 Dec;225(1):89-102. Epub 2006 Jul 15. CHRD, a plant member of the evolutionarily conserved YjgF family, influences photosynthesis and chromoplastogenesis. Leitner-Dagan Y, Ovadis M, Zuker A, Shklarman E, Ohad I, Tzfira T, Vainstein A. The Robert H. Smith Institute of Plant Sciences and Genetics in Agriculture, Faculty of Agricultural, Food and Environmental Quality Sciences, The Hebrew University of Jerusalem, P.O. Box 12, Rehovot 76100, Israel. Studies on the carotenoid-overaccumulating structures in chromoplasts have led to the characterization of proteins termed plastid lipid-associated proteins (PAPs), involved in the sequestration of hydrophobic compounds. Here we characterize the PAP CHRD, which, based on sequence homology, belongs to a highly conserved group of proteins, YER057c/YjgF/UK114, involved in the regulation of basic and vital cellular processes in bacteria, yeast and animals. Two nuclear genes were characterized in tomato plants: one (LeChrDc) is constitutively expressed in various tissues and the other (LeChrDi) is induced by stress in leaves and is upregulated by developmental cues in floral tissues. Using RNAi and antisense approaches, we show their involvement in biologically significant processes such as photosynthesis. The quantum yield of photosynthetic electron flow in transgenic tomato leaves with suppressed LeChrDi/c expression was 30-50% of their control, non-transgenic counterparts and was ascribed to lower PSI activity. Transgenic flowers with suppressed LeChrDi/c also accumulated up to 30% less carotenoids per unit protein as compared to control plants, indicating an interrelationship between PAPs and floral-specific carotenoid accumulation in chromoplasts. We suggest that CHRD's role in the angiosperm reproductive unit may be a rather recent evolutionary development; its original function may have been to protect the plant under stress conditions by preserving plastid functionality. Publication Types: Research Support, Non-U.S. Gov't PMID: 16845531 [PubMed - indexed for MEDLINE] 682: Planta. 2007 Jan;225(2):403-11. Epub 2006 Jul 15. Enhanced Cd2+ -selective root-tonoplast-transport in tobaccos expressing Arabidopsis cation exchangers. Koren'kov V, Park S, Cheng NH, Sreevidya C, Lachmansingh J, Morris J, Hirschi K, Wagner GJ. Department of Plant and Soil Sciences, Plant Biology Program, University of Kentucky, Lexington, KY 40546, USA. Several Arabidopsis CAtion eXchangers (CAXs) encode tonoplast-localized transporters that appear to be major contributors to vacuolar accumulation/sequestration of cadmium (Cd(2+)), an undesirable pollutant ion that occurs in man largely as a result of dietary consumption of aerial tissues of food plants. But, ion-selectivity of individual CAX transporter types remains largely unknown. Here, we transformed Nicotiana tabacum with several CAX genes driven by the Cauliflower Mosaic Virus (CaMV) 35S promoter and monitored divalent cation transport in root-tonoplast vesicles from these plants in order to select particular CAX genes directing high Cd(2+) antiporter activity in root tonoplast. Comparison of seven different CAX genes indicated that all transported Cd(2+), Ca(2+), Zn(2+), and Mn(2+) to varying degrees, but that CAX4 and CAX2 had high Cd(2+) transport and selectivity in tonoplast vesicles. CAX4 driven by the CaMV 35S and FS3 [figwort mosaic virus (FMV)] promoters increased the magnitude and initial rate of Cd(2+)/H(+) exchange in root-tonoplast vesicles. Ion selectivity of transport in root-tonoplast vesicles isolated from FS3::CAX4-expressing plant lines having a range of gene expression was Cd(2+)>Zn(2+)>>Ca(2+)>>Mn(2+) and the ratios of maximal Cd(2+) (and Zn(2+)) versus maximal Ca(2+) and Mn(2+) transport were correlated with the levels of CAX4 expression. Root Cd accumulation in high CAX4 and CAX2 expressing lines was increased in seedlings grown with 0.02 muM Cd. These observations are consistent with a model in which expression of an Arabidopsis-gene-encoded, Cd(2+)-efficient antiporter in host plant roots results in greater root vacuole Cd(2+) transport activity, increased root Cd accumulation, and a shift in overall root tonoplast ion transport selectivity towards higher Cd(2+) selectivity. Results support a model in which certain CAX antiporters are somewhat more selective for particular divalent cations. Publication Types: Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't PMID: 16845524 [PubMed - indexed for MEDLINE] 683: S Afr Med J. 2006 Jun;96(6):509-10. Genetically modified crops--playing a positive role in sustainable development in Africa. Thomson JA. Department of Molecular and Cell Biology, University of Cape Town, South Africa. jat@science.uct.ac.za PMID: 16841131 [PubMed - indexed for MEDLINE] 684: Nat Biotechnol. 2006 Jul;24(7):753. Comment in: Nat Biotechnol. 2006 Nov;24(11):1327-9; author reply 1331-3. Nat Biotechnol. 2006 Nov;24(11):1329-31; author reply 1331-3. Nat Biotechnol. 2006 Nov;24(11):1329; author reply 1331-3. Do cisgenic plants warrant less stringent oversight? Schouten HJ, Krens FA, Jacobsen E. Publication Types: Letter PMID: 16841052 [PubMed - indexed for MEDLINE] 685: Nat Biotechnol. 2006 Jul;24(7):748-9. Regulatory slowdown on GM crop decisions. Jaffe G. Publication Types: Letter PMID: 16841049 [PubMed - indexed for MEDLINE] 686: Nat Biotechnol. 2006 Jul;24(7):735. Elliot Entis. Powell K. Publication Types: News PMID: 16841044 [PubMed - indexed for MEDLINE] 687: Risk Anal. 2006 Jun;26(3):657-70. Examining consumer behavior toward genetically modified (GM) food in Britain. Spence A, Townsend E. RASPH, School of Psychology, University of Nottingham, Nottingham, UK. lpxas@psychology.nottingham.ac.uk This study examined behavior toward genetically modified (GM) food in a British community-based sample. We used an equivalent gain task in which participants actually received the options they chose to encourage truthful responding. In conjunction with this, theory of planned behavior (TPB) components were evaluated so as to examine the relative importance of behavioral influences in this domain. Here, the TPB was extended to include additional components to measure self-identity, moral norms, and emotional involvement. Results indicated that the monetary amounts participants accepted in preference to GM food were significantly lower than those accepted in preference to non-GM food. However, the vast majority of participants were indifferent between GM and non-GM food options. All TPB components significantly predicted behavioral intentions to try GM food, with attitudes toward GM being the strongest predictor. Self-identity and emotional involvement were also found to be significant predictors of behavioral intentions but moral norms were not. In addition, behavioral intentions significantly predicted behavior; however, PBC did not. An additional measure of participants' propensity to respond in a socially desirable manner indicated that our results were not influenced by self-presentation issues, giving confidence to our findings. Overall, it appears that the majority of participants (74.5%) would purchase GM food at some price. Publication Types: Research Support, Non-U.S. Gov't PMID: 16834625 [PubMed - indexed for MEDLINE] 688: Sci Technol Human Values. 2006 Jan;31(1):8-28. Genetic technologies meet the public: the discourses of concern. Lassen J, Jamison A. Department of Human Nutrition, Royal Veterinary and Agricultural University, Copenhagen, Denmark. jlas@kvl.dk To clarify concerns that the public has with genetic technologies, the article presents the results of focus group interviews conducted in Denmark in 2000. The concerns of the public are divided into three ideal-typical categories: social (dealing with environmental and health risks), economic (dealing with both the threats and opportunities of the new technologies), and cultural (taking up ethical and moral concerns). Following a general discussion of why it is important to take these discourses of concern seriously, each discursive category is discussed with examples taken from the focus group interviews. PMID: 16832965 [PubMed - indexed for MEDLINE] 689: Public Aff Q. 2006 Apr;20(2):135-61. Gene patents and Lockean constraints. Shrader-Frechette K. University of Notre Dame, USA. PMID: 16832963 [PubMed - indexed for MEDLINE] 690: Proc Biol Sci. 2006 Aug 7;273(1596):1921-8. Weed seed resources for birds in fields with contrasting conventional and genetically modified herbicide-tolerant crops. Gibbons DW, Bohan DA, Rothery P, Stuart RC, Haughton AJ, Scott RJ, Wilson JD, Perry JN, Clark SJ, Dawson RJ, Firbank LG. RSPB, UK Headquarters, The Lodge, Sandy, Bedfordshire SG19 2DL, UK. david.gibbons@rspb.org.uk The UK Farm Scale Evaluations (FSEs) have shown that the use of broad spectrum herbicides on genetically modified herbicide-tolerant (GMHT) crops can have dramatic effects on weed seed production compared to management of conventional varieties. Here, we use FSE data and information on bird diets to determine how GMHT cropping might change the food resources available to farmland birds. More than 60 fields of each of four crops, spring- and winter-sown oilseed rape, beet and maize, were split, one half being sown with a conventional variety, the other with a GMHT variety. Seed rain from weeds known to be important in the diets of 17 granivorous farmland bird species was measured under the two treatments. In beet and spring oilseed rape, rain of weed seeds important in the diets of 16 bird species was significantly reduced in GMHT compared to conventional halves; for no species did it increase. In winter oilseed rape, rain of weed seeds important in the diets of 10 species was significantly reduced in GMHT halves; for only one species did it increase significantly. By contrast, in maize, rain of weed seeds important in the diets of seven species was significantly greater in GMHT halves; for no species was it reduced. Treatment effects for the total weed seed energy available to each bird species were very similar to those for seed rain alone. Measuring the effects on individual bird species was outside the scope of this study. Despite this, these results suggest that should beet, spring and winter rape crops in the UK be largely replaced by GMHT varieties and managed as in the FSEs, this would markedly reduce important food resources for farmland birds, many of which declined during the last quarter of the twentieth century. By contrast, GMHT maize would be beneficial to farmland birds. Publication Types: Comparative Study Research Support, Non-U.S. Gov't PMID: 16822753 [PubMed - indexed for MEDLINE] 691: Clin Mol Allergy. 2006 Jul 4;4:10. Evaluation of the sensitization rates and identification of IgE-binding components in wild and genetically modified potatoes in patients with allergic disorders. Lee SK, Ye YM, Yoon SH, Lee BO, Kim SH, Park HS. Department of Internal Medicine, College of Medicine, Dong-A University, Busan, Korea. skleeai@daunet.donga.ac.kr BACKGROUND: The potato is one of the most common types of genetically modified (GM) food. However, there are no published data evaluating the impact of genetic manipulations on the allergenicity of GM potatoes. To compare the allergenicity of GM potatoes with that of wild-type potatoes using in vivo and in vitro methods in adult allergy patients sensitized to potatoes. METHODS: A total of 1886 patients with various allergic diseases and 38 healthy controls participated in the study. Skin-prick testing and IgE-ELISA were carried out with extracts prepared from wild-type and GM potatoes. An ELISA inhibition test was used to confirm the binding specificity. IgE-binding components in extracts from the two types of potato were identified by SDS-PAGE and IgE-immunoblotting. The effects of digestive enzymes and heat on the allergenicity of the extracts was evaluated by preincubating the potatoes with or without simulated gastric and intestinal fluids in the absence or presence of heat. RESULTS: Positive responses (ratio of the wheal size induced by the allergen to that induced by histamine (A/H) > or = 2+) to wild-type or GM potato extracts, as demonstrated by the skin-prick test, were observed in 108 patients (5.7%). Serum-specific IgE was detected in 0-88% of subjects who tested positively. ELISA inhibition tests indicated significant inhibition when extract from each type of potato was added. IgE-immunoblot analysis demonstrated the presence of 14 IgE-binding components within the wild-type potato and 9 within the GM potato. Furthermore, a common 45-kDa binding component that yielded similar IgE-binding patterns was noted in more than 80% of the reactions using sera from patients sensitized to wild-type or GM potato. Exposure to simulated gastric fluid and heat treatment similarly inhibited IgE binding by extracts from wild-type and GM potatoes, whereas minimal changes were obtained following exposure of the extracts to simulated intestinal fluid. CONCLUSION: Our results strongly suggest that genetic manipulation of potatoes does not increase their allergenic risk. The sensitization rate of adult allergy patients to both types of extract was 5.7%, and a common major allergen (45 kDa) was identified. PMID: 16817976 [PubMed] 692: Plant Cell Rep. 2006 Dec;25(12):1362-8. Epub 2006 Jun 30. Sequence stability of the T-DNA - plant junctions in tissue culture in Arabidopsis transgenic lines. Papazova N, Windels P, Depicker A, Taverniers I, Roldan-Ruiz I, Milcamps A, Van Bockstaele E, Van Den Eede G, De Loose M. Unit Technology and Food, Institute for Agricultural and Fisheries Research, 9820, Merelbeke, Belgium. nina.papazova@ilvo.vlaanderen.be The stability of the inserted transgenes and particularly the junction regions of transgenic events is a concern of food labeling, traceability and post release monitoring, as these regions are used for development of event-specific DNA-based detection methods. During the standard agricultural breeding practices, the transgenic lines can be exposed to completely different conditions than those in the laboratory environment. Some of these conditions have the potential to affect the stability of the transgenic locus and the surrounding DNA. As tissue culture is recognized as a stressful and mutagenic factor, we have analyzed the effect of this process on the stability of the junction regions at nucleotide level in five Arabidopsis thaliana transgenic lines in comparison with the respective integration loci in ColO and C24 ecotypes. No indication of any kind of alteration at nucleotide level of the junctions was found. The relevance of the stability of the plant-T-DNA junction regions for application of the DNA-based methods in commercial transgenic plants is discussed. Publication Types: Research Support, Non-U.S. Gov't PMID: 16810524 [PubMed - indexed for MEDLINE] 693: BMC Plant Biol. 2006 Jun 26;6:13. Metabolic engineering of potato tuber carotenoids through tuber-specific silencing of lycopene epsilon cyclase. Diretto G, Tavazza R, Welsch R, Pizzichini D, Mourgues F, Papacchioli V, Beyer P, Giuliano G. ENEA, Casaccia Research Center, PO Box 2400, Roma 00100AD, Italy. gianfranco.diretto@casaccia.enea.it BACKGROUND: Potato is a major staple food, and modification of its provitamin content is a possible means for alleviating nutritional deficiencies. beta-carotene is the main dietary precursor of vitamin A. Potato tubers contain low levels of carotenoids, composed mainly of the xanthophylls lutein, antheraxanthin, violaxanthin, and of xanthophyll esters. None of these carotenoids have provitamin A activity. RESULTS: We silenced the first dedicated step in the beta-epsilon- branch of carotenoid biosynthesis, lycopene epsilon cyclase (LCY-e), by introducing, via Agrobacterium-mediated transformation, an antisense fragment of this gene under the control of the patatin promoter. Real Time measurements confirmed the tuber-specific silencing of Lcy-e. Antisense tubers showed significant increases in beta-beta-carotenoid levels, with beta-carotene showing the maximum increase (up to 14-fold). Total carotenoids increased up to 2.5-fold. These changes were not accompanied by a decrease in lutein, suggesting that LCY-e is not rate-limiting for lutein accumulation. Tuber-specific changes in expression of several genes in the pathway were observed. CONCLUSION: The data suggest that epsilon-cyclization of lycopene is a key regulatory step in potato tuber carotenogenesis. Upon tuber-specific silencing of the corresponding gene, beta-beta-carotenoid and total carotenoid levels are increased, and expression of several other genes in the pathway is modified. Publication Types: Research Support, Non-U.S. Gov't PMID: 16800876 [PubMed - indexed for MEDLINE] 694: J Exp Bot. 2006;57(10):2445-53. Epub 2006 Jun 23. Up- and down-regulation of Fragaria x ananassa O-methyltransferase: impacts on furanone and phenylpropanoid metabolism. Lunkenbein S, Salentijn EM, Coiner HA, Boone MJ, Krens FA, Schwab W. Technical University Muenchen, Biomolecular Food Technology, Lise-Meitner-Str. 34, D-85354 Freising, Germany. A complex mixture of hundreds of substances determines strawberry (Fragaria x ananassa) aroma, but only approximately 15 volatiles are considered as key flavour compounds. Of these, 4-hydroxy-2,5-dimethyl-3(2H)-furanone (HDMF) is regarded as the most important, but it is methylated further by FaOMT (Fragaria x ananassa O-methyltransferase) to 2,5-dimethyl-4-methoxy-3(2H)-furanone (DMMF) during the ripening process. It is shown here that transformation of strawberry with the FaOMT sequence in sense and antisense orientation, under the control of the cauliflower mosaic virus 35S promoter, resulted in a near total loss of DMMF, whereas the levels of the other volatiles remained unchanged. FaOMT repression also affected the ratio of feruloyl 1-O-beta-D-glucose and caffeoyl 1-O-beta-D-glucose, indicating a dual function of the enzyme in planta. Thus, FaOMT is involved in at least two different biochemical pathways in ripe strawberry fruit. Publication Types: Research Support, Non-U.S. Gov't PMID: 16798852 [PubMed - indexed for MEDLINE] 695: Microb Cell Fact. 2006 Jun 23;5:23. Live bacterial vaccines--a review and identification of potential hazards. Detmer A, Glenting J. Danish Toxicology Centre, Hørsholm, Denmark. ad@dhigroup.com The use of live bacteria to induce an immune response to itself or to a carried vaccine component is an attractive vaccine strategy. Advantages of live bacterial vaccines include their mimicry of a natural infection, intrinsic adjuvant properties and their possibility to be administered orally. Derivatives of pathogenic and non-pathogenic food related bacteria are currently being evaluated as live vaccines. However, pathogenic bacteria demands for attenuation to weaken its virulence. The use of bacteria as vaccine delivery vehicles implies construction of recombinant strains that contain the gene cassette encoding the antigen. With the increased knowledge of mucosal immunity and the availability of genetic tools for heterologous gene expression the concept of live vaccine vehicles gains renewed interest. However, administration of live bacterial vaccines poses some risks. In addition, vaccination using recombinant bacteria results in the release of live recombinant organisms into nature. This places these vaccines in the debate on application of genetically modified organisms. In this review we give an overview of live bacterial vaccines on the market and describe the development of new live vaccines with a focus on attenuated bacteria and food-related lactic acid bacteria. Furthermore, we outline the safety concerns and identify the hazards associated with live bacterial vaccines and try to give some suggestions of what to consider during their development. PMID: 16796731 [PubMed] 696: Biosci Biotechnol Biochem. 2006 Jun;70(6):1524-7. Recombinant, rice-produced yeast phytase shows the ability to hydrolyze phytate derived from seed-based feed, and extreme stability during ensilage treatment. Hamada A, Yamaguchi K, Harada M, Horiguchi K, Takahashi T, Honda H. Functional Chemicals Laboratory, Mitsui Chemicals, Inc., Togo, Mobara. When fresh rice leaves producing yeast Schwanniomyces occidentalis phytase were grounded and mixed with the whole extract of seed-based feed for pigs, the release of orthophosphate increased significantly. More specifically, phytate, a major source of phosphorus in the seeds, was hydrolyzed by heterologous phytase. Moreover, when transgenic rice plants were ensiled for up to 12 weeks, no decrease in the phytase activity of the heterologous enzyme was observed. This result strongly suggests that transgenic rice plants producing yeast phytase can be stored as silage without any loss of enzyme activity until usage as a feed additive. Publication Types: Research Support, Non-U.S. Gov't PMID: 16794341 [PubMed - indexed for MEDLINE] 697: J AOAC Int. 2006 May-Jun;89(3):893-7. Surface plasmon resonance for detection of genetically modified organisms in the food supply. Gambari R, Feriotto G. Ferrara University, Biotechnology Center, 44100, Ferrara, Italy. gam@dns.unife.it A review is presented demonstrating that biospecific interaction analysis, using surface plasmon resonance (SPR) and biosensor technologies is a simple, rapid, and automatable approach to detect genetically modified organisms (GMOs). Using SPR, we were able to monitor in real-time the hybridization between oligonucleotide or polymerase chain reaction (PCR)-generated probes and target single-stranded PCR products obtained by using as substrates DNA isolated from normal or transgenic soybean and maize. This procedure allows a one-step, nonradioactive detection of GMOs. PCR-generated probes are far more efficient in detecting GMOs than are oligodeoxyribonucleotide probes. This is expected to be a very important parameter, because information on low percentage of GMOs is of great value. Determination of the ability of SPR-based analysis to quantify GMOs should be considered a major research field for future studies, especially for the analyses of food supplies. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 16792091 [PubMed - indexed for MEDLINE] 698: J Agric Food Chem. 2006 Jun 28;54(13):4624-32. Heat-stable phytases in transgenic wheat (Triticum aestivum L.): deposition pattern, thermostability, and phytate hydrolysis. Brinch-Pedersen H, Hatzack F, Stöger E, Arcalis E, Pontopidan K, Holm PB. Research Centre Flakkebjerg, Department of Genetics and Biotechnology, Danish Institute of Agricultural Sciences, DK-4200 Slagelse, Denmark. Henrik.brinchpedersen@agrsci.dk The present paper addresses the question of thermotolerance of in planta synthesized heterologous enzymes using phytase as a model. Two individual transgenic wheat materials expressing an Aspergillus fumigatus phytase with a low denaturation temperature (62.5 degrees C) but a high refolding capacity, and a rationally designed consensus phytase engineered to a high denaturation temperature (89.3 degrees C), were evaluated. High levels of endosperm specific expression were ensured by the wheat high molecular weight glutenin 1DX5 promoter. Immunodetection at the light and electron microscopical level shows unequivocally that the heterologous phytase is deposited in the vacuole, albeit that the transformation constructs were designed for secretion to the apoplast. Evaluation of heat stability properties and kinetic properties unraveled that, under these deposition conditions, heat stability based on high unfolding temperature is superior to high refolding capacity and represents a realistic strategy for improving phosphate and mineral bioavailability in cereal-based feed and food. PMID: 16787007 [PubMed - indexed for MEDLINE] 699: Food Nutr Bull. 2006 Jun;27(2):167-79. Agricultural biodiversity, nutrition, and health: making a difference to hunger and nutrition in the developing world. Frison EA, Smith IF, Johns T, Cherfas J, Eyzaguirre PB. International Plant Genetic Resources Institute, Rome, Italy. e.frison@cgiar.org BACKGROUND: In spite of the strides made globally in reducing hunger, the problems of micronutrient deficiencies and coexisting obesity and related cardiovascular and degenerative diseases constitute a formidable challenge for the future. Attempts to reverse this trend with single-nutrient intervention strategies have met with limited success, resulting in renewed calls for food-based approaches. The deployment of agricultural biodiversity is an approach that entails greater use of local biodiversity to ensure dietary diversity. OBJECTIVE: To outline a new strategy proposed by the International Plant Genetic Resources Institute (IPGRI) that employs agricultural biodiversity as the primary resource for food security and health. METHODS: The authors carried out a meta-analysis to review and assemble existing information on the nutritional and healthful properties of traditional foods based on a diverse set of case studies and food composition and nutritional analysis studies. The methods highlight particular examples of foods where analysis of nutrient and non-nutrient composition reveals important traits to address the growing problems of malnutrition associated with the rise of chronic diseases. Finally, the authors analyze social, economic, and cultural changes that undermine the healthful components of traditional diets. RESULTS: Based on this multidisciplinary and comparative approach, the authors suggest a holistic food-based approach that combines research to assess and document nutritional and healthful properties of traditional foods, investigating options in which nutritionally valuable traditional foods can contribute to better livelihoods, and ways that awareness and promotional campaigns can identify healthful components of traditional diets that fit the needs of urban and market-oriented consumers. CONCLUSIONS: There is an urgent need for agricultural research centers, national agricultural research systems, universities, and community-based organizations to work together under a shared policy framework with the aim of developing a strong evidence base linking biodiversity, nutrition, and health. Although these initiatives are still ongoing, the gains realized in small-scale and local pilot efforts have encouraged IPGRI to work with local partners toward the implementation of scale-up efforts in various regions. Publication Types: Meta-Analysis Review PMID: 16786983 [PubMed - indexed for MEDLINE] 700: Plant Mol Biol. 2006 May;61(1-2):123-39. A microarray-based detection system for genetically modified (GM) food ingredients. Leimanis S, Hernández M, Fernández S, Boyer F, Burns M, Bruderer S, Glouden T, Harris N, Kaeppeli O, Philipp P, Pla M, Puigdomènech P, Vaitilingom M, Bertheau Y, Remacle J. Unité de Recherche en Biologie Cellulaire (URBC), Faculté Universitaire Notre Dame de la Paix, Namur, Belgium. A multiplex DNA microarray chip was developed for simultaneous identification of nine genetically modified organisms (GMOs), five plant species and three GMO screening elements, i.e. the 35S promoter, the nos terminator and the nptII gene. The chips also include several controls, such as that for the possible presence of CaMV. The on-chip detection was performed directly with PCR amplified products. Particular emphasis was placed on the reduction of the number of PCR reactions required and on the number of primers present per amplification tube. The targets were biotin labelled and the arrays were detected using a colorimetric methodology. Specificity was provided by specific capture probes designed for each GMO and for the common screening elements. The sensitivity of the assay was tested by experiments carried out in five different laboratories. The limit of detection was lower than 0.3% GMO for all tests and in general around 0.1% for most GMOs. The chip detection system complies with the requirements of current EU regulations and other countries where thresholds are established for the labelling of GMO. Publication Types: Evaluation Studies Research Support, Non-U.S. Gov't PMID: 16786296 [PubMed - indexed for MEDLINE] 701: Plant Mol Biol. 2006 May;61(1-2):47-62. Spatial organisation of four enzymes from Stevia rebaudiana that are involved in steviol glycoside synthesis. Humphrey TV, Richman AS, Menassa R, Brandle JE. Agriculture and Agri-Food Canada, Southern Crop Protection and Food Research Centre, London, Ont. The sweet steviol glycosides found in the leaves of Stevia rebaudiana Bert. are derived from the diterpene steviol which is produced from a branch of the gibberellic acid (GA) biosynthetic pathway. An understanding of the spatial organisation of the two pathways including subcellular compartmentation provides important insight for the metabolic engineering of steviol glycosides as well as other secondary metabolites in plants. The final step of GA biosynthesis, before the branch point for steviol production, is the formation of (-)-kaurenoic acid from (-)-kaurene, catalysed by kaurene oxidase (KO). Downstream of this, the first committed step in steviol glycoside synthesis is the hydroxylation of kaurenoic acid to form steviol which is then sequentially glucosylated by a series of UDP-glucosyltransferases (UGTs) to produce the variety of steviol glycosides. The subcellular location of KO and three of the UGTs involved in steviol glycoside biosynthesis was investigated by expression of GFP fusions and cell fractionation which revealed KO to be associated with the endoplasmic reticulum and the UGTs in the cytoplasm. It has also been shown by expressing the Stevia UGTs in Arabidopsis that the pathway can be partially reconstituted by recruitment of a native Arabidopsis glucosyltransferase. PMID: 16786291 [PubMed - indexed for MEDLINE] 702: Transgenic Res. 2006 Jun;15(3):277-89. Mycotoxin reduction in Bt corn: potential economic, health, and regulatory impacts. Wu F. Environmental, Occupational Health, Graduate School of Public Health, University of Pittsburgh, 130 DeSoto St., Pittsburgh, PA 15261, USA. fwu@eoh.pitt.edu Genetically modified (GM) Bt corn, through the pest protection that it confers, has lower levels of mycotoxins: toxic and carcinogenic chemicals produced as secondary metabolites of fungi that colonize crops. In some cases, the reduction of mycotoxins afforded by Bt corn is significant enough to have an economic impact, both in terms of domestic markets and international trade. In less developed countries where certain mycotoxins are significant contaminants of food, Bt corn adoption, by virtue of its mycotoxin reduction, may even improve human and animal health. This paper describes an integrated assessment model that analyzes the economic and health impacts of two mycotoxins in corn: fumonisin and aflatoxin. It was found that excessively strict standards of these two mycotoxins could result in global trade losses in the hundreds of millions US dollars annually, with the US, China, and Argentina suffering the greatest losses. The paper then discusses the evidence for Bt corn's lower levels of contamination of fumonisin and aflatoxin, and estimates economic impacts in the United States. A total benefit of Bt corn's reduction of fumonisin and aflatoxin in the US was estimated at 23 million dollars annually. Finally, the paper examines the potential policy impacts of Bt corn's mycotoxin reduction, on nations that are making a decision on whether to allow commercialization of this genetically modified crop. PMID: 16779644 [PubMed - indexed for MEDLINE] 703: J Zhejiang Univ Sci B. 2006 Jul;7(7):591-5. Erratum in: J Zhejiang Univ Sci B. 2007 Feb;8(2):115. Characteristics of transgenic tomatoes antisensed for the ethylene receptor genes LeETR1 [corrected] and LeETR2 [corrected] Wang ZF, Ying TJ, Zhang Y, Bao BL, Huang XD. School of Biosystem Engineering and Food Science, Zhejiang University, Hangzhou 310029, China. Two stable transformed lines containing antisense LeETR1 [corrected] or LeETR2 [corrected] sequences and their hybridized line were investigated to determine the effect of LeETR1 [corrected] and LeETR2 [corrected] specificity in the ethylene receptor family in tomato (Lycopersicon esculentum Mill.) on ethylene signaling. The transgenic line ale1 containing antisense LeETR1 [corrected] displayed shorter length of seedling grown in the dark and adult plant in the light, severe epinastic petiole, and accelerated abscission of petiole explant and senescence of flower explant, compared with its wild type B1. The transgenic line ale2 containing antisense LeETR2 [corrected] also exhibited shorter hypocotyls and slightly accelerated abscission. The phenotypes of cross line dale of LeETR1 [corrected] and LeETR2 [corrected] were close to ale1 in many aspects. These results suggested that LeETR1 [corrected] probably plays a relatively important role in ethylene signaling of tomato growth and development. Publication Types: Research Support, Non-U.S. Gov't PMID: 16773735 [PubMed - indexed for MEDLINE] 704: Plant Physiol. 2006 Aug;141(4):1306-15. Epub 2006 Jun 9. Glyphosate-induced anther indehiscence in cotton is partially temperature dependent and involves cytoskeleton and secondary wall modifications and auxin accumulation. Yasuor H, Abu-Abied M, Belausov E, Madmony A, Sadot E, Riov J, Rubin B. R.H. Smith Institute of Plant Sciences and Genetics in Agriculture, Faculty of Agricultural, Food, and Environmental Sciences, The Hebrew University of Jerusalem, Rehovot 76100, Israel. yasuor@agri.huji.ac.il Yield reduction caused by late application of glyphosate to glyphosate-resistant cotton (Gossypium hirsutum; GRC) expressing CP4 5-enol-pyruvylshikmate-3-P synthase under the cauliflower mosaic virus-35S promoter has been attributed to male sterility. This study was aimed to elucidate the factors and mechanisms involved in this phenomenon. Western and tissue-print blots demonstrated a reduced expression of the transgene in anthers of GRC compared to ovules of the same plants. Glyphosate application to GRC grown at a high temperature regime after the initiation of flower buds caused a complete loss of pollen viability and inhibition of anther dehiscence, while at a moderate temperature regime only 50% of the pollen grains were disrupted and anther dehiscence was normal. Glyphosate-damaged anthers exhibited a change in the deposition of the secondary cell wall thickenings (SWT) in the endothecium cells, from the normal longitudinal orientation to a transverse orientation, and hindered septum disintegration. These changes occurred only at the high temperature regime. The reorientation of SWT in GRC was accompanied by a similar change in microtubule orientation. A similar reorientation of microtubules was also observed in Arabidopsis (Arabidopsis thaliana) seedlings expressing green fluorescent protein tubulin (tubulin alpha 6) following glyphosate treatment. Glyphosate treatment induced the accumulation of high levels of indole-3-acetic acid in GRC anthers. Cotton plants treated with 2,4-dichlorophenoxyacetic acid had male sterile flowers, with SWT abnormalities in the endothecium layer similar to those observed in glyphosate-treated plants. Our data demonstrate that glyphosate inhibits anther dehiscence by inducing changes in the microtubule and cell wall organization in the endothecium cells, which are mediated by auxin. Publication Types: Research Support, Non-U.S. Gov't PMID: 16766672 [PubMed - indexed for MEDLINE] 705: Toxicol Lett. 2006 Sep 10;165(3):250-6. Epub 2006 May 7. Mutations induced by carcinogenic doses of aristolochic acid in kidney of Big Blue transgenic rats. Chen L, Mei N, Yao L, Chen T. Division of Genetic and Reproductive Toxicology, National Center for Toxicological Research, Food and Drug Administration, HFT-130, 3900 NCTR Road, Jefferson, AR 72079, USA. Aristolochic acid (AA) is present in at least 65 different kinds of plants, many of which are used as herbal folk remedies. AA is considered one of the most potent plant carcinogens in humans and animals. It has been associated with the development of urothelial cancers in humans, and kidney and forestomach tumors in rats. In the present study, we used the Big Blue transgenic rat model to evaluate the mutagenicity of AA in kidney of rats and to define the mechanism of action for the tumor induction by AA. Groups of six male Big Blue transgenic rats were gavaged with 0, 0.1, 1.0 and 10.0mgAA/kg body weight 5 times a week for 12 weeks, a treatment protocol that resulted in tumors in kidneys and other tissues. The animals were sacrificed 1 day after the final treatment and the kidneys were isolated for assays to determine the mutant frequencies (MFs) and types of mutations induced by AA in the transgenic cII gene. AA treatment resulted in a strong linear relationship between MF inductions and treatment dose (R(2)=0.998). The cII MFs were 29+/-6x10(-6), 78+/-21x10(-6), 242+/-104x10(-6) and 1319+/-360x10(-6) in the control, low, medium and high dose treatment groups, respectively (p<0.001 for all pair wise comparisons among the four treatment groups). These MFs correlated strongly with tumor incidences induced by the different doses of AA (Mengs et al., 1982). Sequence analysis of the cII mutants revealed that there was a statistically significant difference between the mutational spectra in the AA-treated and control rats (p<0.05). A:T-->T:A transversion was the predominant type of mutation in the AA-treated rats whereas G:C-->A:T transition was the main type of mutations in the control rats. These results suggest that AA induces kidney tumors in rats though a mutagenic mechanism of action. Publication Types: Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. PMID: 16764999 [PubMed - indexed for MEDLINE] 706: Mol Nutr Food Res. 2006 Jul;50(7):645-54. Allergen-specific IgE testing in the diagnosis of food allergy and the event of a positive match in the bioinformatics search. van Ree R, Vieths S, Poulsen LK. Academic Medical Center, Amsterdam, The Netherlands. Current documents on risk assessment of genetically modified foods recommend including IgE-binding tests on sera from allergic patients. However, there is no generally accepted recommendation on technical aspects of the testing procedures or on the interpretation of the results, despite that fact that both false positive and false-negative results may be caused by variability of the test procedures. The present article discusses the state-of-the-art of serological test procedures for qualitative and quantitative determination of specific IgE and interpretation of test results. It is emphasized that the use of sera from clinically well-characterized subjects is of high importance. In the case of a positive test result, the biological activity of the detected IgE antibodies, i. e., the potential to trigger mediator release from basophils or mast cells in an allergen-specific manner, should be taken into account. However, present data also indicate that validation of such mediator release tests is required, both in terms of experimental protocols and with respect to correlation of the test results with the clinical situation. Further studies are also required to prove the usefulness of targeted serum screening, i. e., the testing of gene products from organisms not known to be allergenic with sera from subjects allergic to related species. Publication Types: Review PMID: 16764014 [PubMed - indexed for MEDLINE] 707: Foodborne Pathog Dis. 2006 Summer;3(2):157-62. Food safety--who is responsible? Rollin BE. Department of Philosophy, Colorado State University, Fort Collins, Colorado 80523-1781, USA. Bernard.Rollin@colostate.edu Though scientists believe that issues of risk can be handled without appeal to values in general or ethics in particular, this is demonstrably false. The very notion of risk is enmeshed in a complex of social ethics. This is clearly true with regard to food safety. With this in mind, it is plausible to affirm that responsibility for food safety at a given point in the chain from producer to consumer rests with the person or entity under whose control the management of that risk most plausibly lies. This principle is illustrated with various examples and with clear cases of industry shouldering and avoiding responsibility. An additional ethical concern relevant to food safety arises from genetically modified foods. Given that the situation here is uncertain and risk unknown, it is hard to see who is responsible for managing such risks. It is arguable that this situation militates in favor of labeling, since consumers are in effect research subjects. The reasonable moral approach to risk we have outlined is jeopardized by the societal tendency towards "victimology" and abrogation of personal responsibility. In such a world, it is incumbent on industry to educate the public with regard to consumer minimization of food safety risks, the impossibility of zero-risk situations, and the economic costs to freedom of protectionism. PMID: 16761941 [PubMed - indexed for MEDLINE] 708: J Environ Sci Health B. 2006;41(4):437-49. Gene transfer and cauliflower mosaic virus promoter 35S activity in mammalian cells. Paparini A, Romano-Spica V. IUSM, University Institute for Movement Science, Department of Health Sciences, Rome, Italy. The cauliflower mosaic virus 35S promoter (CaMV35s) is extensively used in genetically modified crops for human and animal consumption. Horizontal gene transfer is attracting particular attention, in light of experimental reports, showing the presence of dietary DNA into animal tissues. Health implications may derive from possible activities of the heterologous promoter in mammalian cells after integration in the host genome. To evaluate this hypothesis, in vivo and in vitro experiments were performed using GFP as reporter gene. Recombinant plasmid DNA was fed to Balb/c mice and searched in several tissues by PCR amplification. The activity of the plant virus promoter was assessed by RT-PCR and fluorescence microscopy after liposome-mediated transfection of murine gonadic cells. Obtained data did not highlight evidences of dietary DNA transfer in mice. No CaMV35s transcriptional activity was detected in this experimental model. These findings emphasize the need for further studies and standardized methods. Publication Types: Research Support, Non-U.S. Gov't PMID: 16753962 [PubMed - indexed for MEDLINE] 709: Nat Biotechnol. 2006 Jun;24(6):713-7. Epub 2006 Jun 4. Herbicide-resistance conferred by expression of a catalytic antibody in Arabidopsis thaliana. Weiss Y, Shulman A, Ben Shir I, Keinan E, Wolf S. The Institute of Plant Sciences and Genetics in Agriculture and the Otto Warburg Minerva Center for Agricultural Biotechnology, Faculty of Agricultural, Food and Environmental Quality Sciences, The Hebrew University, Rehovot, Israel. Engineering herbicide resistance in crops facilitates control of weed species, particularly those that are closely related to the crop, and may be useful in selecting lines that have undergone multiple transformation events. Here we show that herbicide-resistant plants can be engineered by designing an herbicide and expressing a catalytic antibody that destroys the herbicide in planta. First, we developed a carbamate herbicide that can be catalytically destroyed by the aldolase antibody 38C2. This compound has herbicidal activity on all three plant species tested. Second, the light chain and half of the heavy chain (Fab) of the catalytic antibody were targeted to the endoplasmic reticulum in two classes of Arabidopsis thaliana transformants. Third, the two transgenic plants were crossed to produce an herbicide-resistant F1 hybrid. The in vitro catalytic activity of the protein from F1 hybrids corroborates that catalytic antibodies can be constitutively expressed in transgenic plants, and that they can confer a unique trait. Publication Types: Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. PMID: 16751769 [PubMed - indexed for MEDLINE] 710: Mol Nutr Food Res. 2006 Jul;50(7):604-9. Review of the development of methodology for evaluating the human allergenic potential of novel proteins. Taylor SL. University of Nebraska, Food Allergy Research & Resource Program, Lincoln, Nebraska 68583, USA. staylor2@unl.edu Safety assessment of novel proteins in genetic-engineered foods is a key component of the overall safety evaluation for these products. Since allergens are typically proteins, assessment of the potential allergenicity of the novel proteins in genetically engineered foods is critical. This article reviews methods available to assess the potential allergenicity of novel proteins, as well as problems and deficiencies in the existing methods. The role of bioinformatics and knowledge of allergenic epitopes in developing new approaches to this problem is discussed. Publication Types: Comparative Study Review PMID: 16736463 [PubMed - indexed for MEDLINE] 711: Plant Foods Hum Nutr. 2006 Mar;61(1):23-8. Mineral profile and variability in vegetable amaranth (Amaranthus tricolor). Shukla S, Bhargava A, Chatterjee A, Srivastava J, Singh N, Singh SP. Division of Genetics and Plant Breeding, National Botanical Research Institute, Rana Pratap Marg, Lucknow, 226001, India. s_shukla31@rediffmail.com Populations in North India depend on a number of vegetable crops of which Amaranthus spp. is the most important since it is the only crop available in the hot summer months when no other foliage crop grows in the field. However, reports on mineral composition of leaves are rare with absolutely no information on the qualitative improvement of foliage yield with special reference to minerals. Studies on correlation among the minerals as well as with yield and leaf attributes are also lacking. Hence, we report the proximate mineral composition in 30 strains of A. tricolor along with some suggestions for qualitative improvement of the foliage yield with reference to minerals. Our study showed that vegetable amaranth is a rich source of minerals like calcium (1.7 +/- 0.04 g/100 g), iron (1233.8 +/- 50.02 mg/kg), and zinc (791.7 +/- 28.98 mg/kg). The heritability estimates were high for most of the traits, with potassium and calcium showing high values, while comparatively lower values were recorded for magnesium and nickel. Nickel was the only mineral that showed positive correlation with all the minerals, as well as with leaf size and foliage yield. Zinc showed strong positive relationship with iron (0.66**) and manganese (0.74**), and was the only mineral exhibiting significant positive association with foliage yield. This study would be of use in enhancement of selected minerals in different regions according to local preferences and nutrient deficiency prevalent among the populations. PMID: 16736385 [PubMed - indexed for MEDLINE] 712: Shokuhin Eiseigaku Zasshi. 2006 Apr;47(2):J185-8. [Codex ad hoc Intergovernmental Task Force on Foods Derived from Biotechnology] [Article in Japanese] Umeda T. Department of Food Safety, Ministry of Health, Labour and Welfare, Tokyo, Japan. PMID: 16729673 [PubMed - indexed for MEDLINE] 713: J Biosci Bioeng. 2006 Mar;101(3):203-11. Immunogenic and allergenic potentials of natural and recombinant innocuous proteins. Matsuda T, Matsubara T, Hino S. Department of Applied Molecular Biosciences, Graduate School of Bioagricultural Sciences, Nagoya University, Chikusa, Nagoya 464-8601, Japan. tmatsuda@agr.nagoya-u.ac.jp A new aspect of protein immunogenic and allergenic properties has become important recently, when there is a higher chance that our immune system will be exposed to novel protein antigens and/or familiar protein antigens with an unprecedented high frequency and large amount. These proteins are innocuous, nontoxic, and noninvasive by themselves, and include various natural proteins from the environment and recombinant proteins from industry. The technical term allergenic has been used for such proteins and their abilities to induce specific IgE production and to cross-link IgE/Fc epsilonRI on the surface of mast cells and basophiles have been recognized. As for the environmental proteins, some physicochemical properties (solubility, stability, and permeability across a mucosal epithelium) of the proteins indirectly play important roles in their allergenic potential because they do not originate from invasive pathogens as vehicles. Indeed, several lines of experimental evidences have been accumulated indicating that all proteins are absorbed across mucosal epithelia by transcellular transport and/or through interstitial spaces among the epithelial cells but not at equal levels. Some animal models have been established for natural sensitization to some allergenic proteins by feeding or intragastric administration without an adjuvant and, in a few cases, some symptoms resembling human allergy and even anaphylaxis have been induced by oral challenge with the proteins. Sometimes, even to self-proteins, the immunogenic or allergenic potential is given by post-translational modifications and possibly by unknown structural/conformational alterations, when they are exogenous self-proteins, such as recombinant human proteins for drug use. Despite the accumulation of knowledge and the progress in analytical technology on protein allergenicity, it is still crucial to predict the allergenic potential of novel and unused proteins. However, some animal models are applicable for assessing the relative allergenic potential of processed proteins in comparison with that of native proteins in preclinical studies. Publication Types: Review PMID: 16716919 [PubMed - indexed for MEDLINE] 714: Trends Plant Sci. 2006 Jun;11(6):261-3. Epub 2006 May 11. Wild sex in the grasses. Able JA, Langridge P. Molecular Plant Breeding Cooperative Research Centre, School of Agriculture, Food & Wine, The University of Adelaide, Glen Osmond, SA 5064, Australia. jason.able@adelaide.edu.au To date, alien introgression of agronomically important traits into bread wheat (Triticum aestivum) from wild relatives has not been readily achievable through traditional breeding practices. However, this door might now be unlocked. The insightful research published recently by Graham Moore and his team delivers a likely candidate in the form of a cdc2-kinase-related gene family for the Ph1 locus--a chromatin region located on chromosome 5B that is responsible for homologous chromosome pairing integrity in bread wheat. PMID: 16697246 [PubMed - indexed for MEDLINE] 715: Virology. 2006 Aug 1;351(2):455-65. Epub 2006 May 11. Functional analysis of the 5' untranslated region of potexvirus RNA reveals a role in viral replication and cell-to-cell movement. Lough TJ, Lee RH, Emerson SJ, Forster RL, Lucas WJ. Horticulture and Food Research Institute of New Zealand, Plant Health and Development Group, Private Bag 11030, Palmerston North, New Zealand. t.lough@genesis.co.nz Cell-to-cell movement of potexviruses requires cognate recognition between the viral RNA, the triple gene block proteins (TGBp1-3) and the coat protein (CP). cis-acting motifs required for recognition and translocation of viral RNA were identified using an artificial potexvirus defective RNA encoding a green fluorescent protein (GFP) reporter transcriptionally fused to the terminal viral sequences. Analysis of GFP fluorescence produced in vivo from these defective RNA constructs, referred to as chimeric RNA reporters, was used to identify viral cis-acting motifs required for RNA trafficking. Mapping experiments localized the cis-acting element to nucleotides 1-107 of the Potato virus X (PVX) genome. This sequence forms an RNA secondary structural element that has also been implicated in viral plus-strand accumulation [Miller, E.D., Plante, C.A., Kim, K.-H., Brown, J.W. and Hemenway, C. (1998) J. Mol. Biol. 284, 591-608]. While replication and movement functions associated with this region have not been separated, these results are consistent with sequence-specific recognition of RNA by the viral movement protein(s). This situation is unusual among viral movement proteins that typically function to translocate RNA between cells in a non-sequence-specific manner. These data support the concept of cis-acting elements specifying intercellular potexvirus RNA movement and thus provide a basis for dissection of RNA-mediated intercellular communication in plants. Publication Types: Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. PMID: 16697024 [PubMed - indexed for MEDLINE] 716: Physiol Behav. 2006 Nov 30;89(4):465-71. Epub 2006 May 11. Amylinergic control of food intake. Lutz TA. Institute of Veterinary Physiology, Vetsuisse Faculty University of Zurich and Center of Integrative Human Physiology, Winterthurerstrasse 260, 8057 Zurich, Switzerland. tomlutz@vetphys.unizh.ch Amylin is a pancreatic B-cell hormone that plays an important role in the regulation of nutrient fluxes. As such, amylin reduces food intake in laboratory animals and man, slows gastric emptying and it reduces postprandial glucagon secretion. Amylin deficiency which occurs concomitantly to insulin deficiency in diabetes mellitus, may therefore contribute to some of the major derangements associated with this disorder (hyperphagia, excessive glucagon secretion, accelerated rate of gastric emptying). The described actions of amylin all seem to depend on a direct effect of amylin on the area postrema (AP). As to amylin's satiating effect, the physiological relevance of this action is underlined by studies involving specific amylin antagonists and amylin-deficient mice. In the AP, amylin seems to modulate the anorectic signal elicited by CCK. Subsequent to AP activation, the amylin signal is conveyed to the forebrain via distinct relay stations. Within the lateral hypothalamic area, amylin diminishes the expression of orexigenic neuropeptides such as orexin and MCH. Whether these effects contribute to amylin's short term satiating action remains to be determined. Recent studies suggest that amylin may also play a role as a long-term, lipostatic signal, especially when other feedback systems to the brain are deficient. Obese, leptin-resistant Zucker rats which are hyperinsulinemic and hyperamylinemic, were chronically infused with the amylin antagonist AC 187. AC 187 significantly elevated food intake in obese Zucker rats while having no effect in lean controls. This indicates that at least under certain conditions, chronic blockade of endogenous amylin action may lead to an increase in food intake and/or body weight. As mentioned, the site and mechanism of action for peripheral amylin to reduce food intake seems to be well established. It is less clear how centrally administered amylin reduces food intake although it is well known that 3rd ventricular administration of amylin produces a very strong and long-lasting anorectic action. Amylin receptors have been described in various hypothalamic nuclei but the endogenous ligand of these receptors remains to be investigated. The same holds true as to the physiological relevance of the anorectic effect seen after central amylin administration. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 16697020 [PubMed - indexed for MEDLINE] 717: Nature. 2006 May 11;441(7090):149. Challengers in the field. Macilwain C. PMID: 16688145 [PubMed - indexed for MEDLINE] 718: Trends Biotechnol. 2006 Jul;24(7):305-11. Epub 2006 May 6. Genetic engineering of wheat--current challenges and opportunities. Bhalla PL. Plant Molecular Biology and Biotechnology Laboratory, Australian Research Council Centre of Excellence for Integrative Legume Research, The University of Melbourne, Parkville, Victoria 3010, Australia. premlb@unimelb.edu.au Wheat is one of the major staple food crops grown worldwide; however, productivity in cereal crops has not kept pace with the world population growth. A significant increase in wheat production (>40% by 2020) is needed simply to keep up with the growing demand. This increase is unlikely to be achieved by conventional plant breeding methods because of the limited gene pool available. The application of recombinant techniques to improve wheat quality and yield is not only desirable but also has potential to open up new opportunities. Although there has been significant progress in developing gene-transformation technologies for improving these traits, this remains an important challenge for plant biotechnology. Obstacles to translate the full potential of the genomic era to wheat breeding include the need to develop elite wheat varieties without selectable markers, introducing minimal or nil intergenic DNA and social and market issues concerning genetically engineered food products. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 16682090 [PubMed - indexed for MEDLINE] 719: Hist Cienc Saude Manguinhos. 2000 Jul-Oct;7(2):493-8. [Transgenics without Manichaeism] [Article in Portuguese] Valle S. Fundação Oswaldo Cruz. We live in an era characterized by the hegemony of science and technology, an era fraught with questions awaiting answers which would enable a safe and sustainable future for humankind. The development of agro-industrial processes - food products in particular - through recombinant DNA technology has enhanced the profit prospects of the few big biotechnology companies and of large-scale farmers who have access to the latest technological developments. We thus oppose a moratorium on recombinant DNA technology. Moreover, hasty statements about risk-free transgenics may be misleading in the absence of extensive safety tests. There is a pressing need for the establishment of biosafety policy in this country involving the organized civil society and every government agency responsible for monitoring such matters. There is also the need to put in place a bio-surveillance and a code of ethics regarding genetic manipulation. Publication Types: English Abstract Historical Article PMID: 16680900 [PubMed - indexed for MEDLINE] 720: Hist Cienc Saude Manguinhos. 2000 Jul-Oct;7(2):481-91. [Implications of transgenics for environmental and agricultural sustainability] [Article in Portuguese] Nodari RO, Guerra MP. Departamento de Fitotecnia, Universidade Federal de Santa Catarina, Florianópolis, Brasil. The potential risks of GMOs, their impact on human and animal health, and on the environment, as well as their socioeconomic effects, have generated a worldwide discussion which is far from drawing to a close for lack of sufficient information. Part of this information supports risk-hypotheses previously put forward. Thus the presence of transgenic plant genes in other plants and in other organisms has been confirmed in several occasions. Therefore, gene dissemination to plants of the same species as well as to widely different species is already regarded as an actual risk. The principle of substantial equivalence has opened the way for the liberation of transgenic plants for commercial crops, despite short-term tests, which are quantitatively and qualitatively insufficient to certify that the foods deriving from those plants are healthy and safe. Thus, the adoption of the so-called precautionary principle (PP) has turned out to be the most adequate safety measure to date, or else until scientific data should be able to demonstrate the actual impact of transgenic plants on human and animal health, and on the environment. Publication Types: English Abstract Historical Article PMID: 16680899 [PubMed - indexed for MEDLINE] 721: Hist Cienc Saude Manguinhos. 2000 Jul-Oct;7(2):465-79. [A technology with multiple applications] [Article in Portuguese] Pinheiro MM, Gerhardt L, Margis R. Universidade Estadual do Rio de Janeiro. Plant breeding has been a human practice for some thousands of years. However, this process of domestication has made plants more vulnerable to pests and diseases. Classic plant breeding has allowed the genetic manipulation of plants through crossings with a resulting increase in crop productivity. Recently, the recombinant DNA technology has increased the possibilities of integration of exogenous genes to the plant genome, resulting in the production of transgenic plants. Despite the great debate on this issue, such plants represent to date a promising avenue for plant breeding. There are many examples of gene transference strategies which have been successful in promoting resistance to herbicides, viruses, fungi, bacteria and insects, or in producing an increase in food quality. In addition to biotechnological applications, transgenic plants have made a significant contribution to the study of gene functioning, such as the analysis of genic expression regulation and the study of protein functions codified by distinct plant genes. Publication Types: English Abstract Historical Article PMID: 16680898 [PubMed - indexed for MEDLINE] 722: Hist Cienc Saude Manguinhos. 2000 Jul-Oct;7(2):437-9. [GMOs on the menu: crack them before eating!] [Article in Portuguese] Massarani L. Publication Types: Historical Article PMID: 16680894 [PubMed - indexed for MEDLINE] 723: Nat Biotechnol. 2006 May;24(5):498; author reply 499. Comment in: Nat Biotechnol. 2007 Jun;25(6):624-6. Comment on: Nat Biotechnol. 2005 Nov;23(11):1348-9. Transgenic plant science priorities. Pelletier D. Publication Types: Comment Letter PMID: 16680123 [PubMed - indexed for MEDLINE] 724: Nat Biotechnol. 2006 May;24(5):481. US-Indian agbiotech deal under scrutiny. Jayaraman KS. Publication Types: News PMID: 16680115 [PubMed - indexed for MEDLINE] 725: Biomed Environ Sci. 2006 Feb;19(1):42-6. Comparison of ileal digested production of parental rice and rice genetically modified with cowpeas trypsin inhibitor. Han JH, Yang YX, Men JH, Bian LH, Guo J. Institute of Nutrition and Food Safety, Chinese Center for Disease Control and Prevention, 27 Nanwei Road, Beijing 100050, China. OBJECTIVE: To compare the ileal digestibility of protein and amino acids in parental rice and rice genetically modified with sck gene. METHODS: Six experimental swines were surgically fixed with a simple T-cannula at the terminal ileum and fed with parental rice and rice genetically modified with sck gene alternately. The ileum digesta were collected and analyzed for determination of apparent and true digestibility of protein and amino acids. RESULTS: The apparent and true digestibility of protein was similar in these two types of rice. Except for the apparent digestibility of lysine, there was no difference in the apparent and true digestibility of the other 17 amino acids. CONCLUSION: The digestibility of protein and amino acids is not changed by the insertion of foreign gene, so it can meet the request of "substantial equivalence" in digestibility of protein and amino acids. Publication Types: Comparative Study PMID: 16673817 [PubMed - indexed for MEDLINE] 726: Proc Nutr Soc. 2006 May;65(2):198-203. Plant foods for human health: research challenges. Mathers JC. Human Nutrition Research Centre, School of Clinical Medical Sciences, University of Newcastle, Newcastle upon Tyne NE2 4HH, UK. john.mathers@ncl.ac.uk Plants provide the major part of human food intake. Whilst advances in agronomic characteristics (improved yield and better pest and disease resistance) continue to be a very high priority, there is increasing opportunity to enhance the nutritional value of plant based diets by improving the nutritional quality of staple foods. We now have proof of principle that genetic engineering can be used to produce plant-derived human vaccines. In relation to plant foods for human health, the research challenges include understanding: (1) why certain foods cause adverse reactions in some individuals but not in others; (2) the mechanisms of action of apparently 'protective' foods such as fruits and vegetables. There is also a need to develop much more informative and robust methods for measuring dietary exposure to specific plant foods or food constituents, including both recent exposure, for which a metabolomics approach may be particularly helpful, and long-term exposure. Publication Types: Lectures PMID: 16672081 [PubMed - indexed for MEDLINE] 727: Proc Nutr Soc. 2006 May;65(2):169-81. Biofortification of UK food crops with selenium. Broadley MR, White PJ, Bryson RJ, Meacham MC, Bowen HC, Johnson SE, Hawkesford MJ, McGrath SP, Zhao FJ, Breward N, Harriman M, Tucker M. Plant Sciences Division, Univerisity of Nottingham, Sutton Bonington, Loughborough LE12 5RD, UK. martin.broadley@nottingham.ac.uk Se is an essential element for animals. In man low dietary Se intakes are associated with health disorders including oxidative stress-related conditions, reduced fertility and immune functions and an increased risk of cancers. Although the reference nutrient intakes for adult females and males in the UK are 60 and 75 microg Se/d respectively, dietary Se intakes in the UK have declined from >60 microg Se/d in the 1970s to 35 microg Se/d in the 1990s, with a concomitant decline in human Se status. This decline in Se intake and status has been attributed primarily to the replacement of milling wheat having high levels of grain Se and grown on high-Se soils in North America with UK-sourced wheat having low levels of grain Se and grown on low-Se soils. An immediate solution to low dietary Se intake and status is to enrich UK-grown food crops using Se fertilisers (agronomic biofortification). Such a strategy has been adopted with success in Finland. It may also be possible to enrich food crops in the longer term by selecting or breeding crop varieties with enhanced Se-accumulation characteristics (genetic biofortification). The present paper will review the potential for biofortification of UK food crops with Se. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 16672078 [PubMed - indexed for MEDLINE] 728: Proc Nutr Soc. 2006 May;65(2):153-9. Biofortification of essential nutritional compounds and trace elements in rice and cassava. Sautter C, Poletti S, Zhang P, Gruissem W. Institute of Plant Science, Swiss Federal Institute of Technology Zurich, Universitaetsstr. 2, CH-8092 Zurich, Switzerland. Christof.Sautter@ipw.biol.ethz.ch Plant biotechnology can make important contributions to food security and nutritional improvement. For example, the development of 'Golden Rice' by Professor Ingo Potrykus was a milestone in the application of gene technology to deliver both increased nutritional qualities and health improvement to wide sections of the human population. Mineral nutrient and protein deficiency as well as food security remain the most important challenges for developing countries. Current projects are addressing these issues in two major staple crops, cassava (Manihot esculenta Crantz) and rice. The tropical root crop cassava is a major source of food for approximately 600 million of the population worldwide. In sub-Saharan Africa >200 million of the population rely on cassava as their major source of dietary energy. The nutritional quality of the cassava root is not sufficient to meet all dietary needs. Rice is the staple food for half the world population, providing approximately 20% of the per capita energy and 13% of the protein for human consumption worldwide. In many developing countries the dietary contributions of rice are substantially greater (29.3% dietary energy and 29.1% dietary protein). The current six most popular 'mega' rice varieties (in terms of popularity and acreage), including Chinese hybrid rice, have an incomplete amino acid profile and contain limited amounts of essential micronutrients. Rice lines with improved Fe contents have been developed using genes that have functions in Fe absorption, translocation and accumulation in the plant, as well as improved Fe bioavailability in the human intestine. Current developments in biotechnology-assisted plant improvement are reviewed and the potential of the technology in addressing human nutrition and health are discussed. Publication Types: Review PMID: 16672076 [PubMed - indexed for MEDLINE] 729: Plant Mol Biol. 2006 Mar;60(5):679-98. "HAIRY CANOLA"--Arabidopsis GL3 induces a dense covering of trichomes on Brassica napus seedlings. Gruber MY, Wang S, Ethier S, Holowachuk J, Bonham-Smith PC, Soroka J, Lloyd A. Agriculture and Agri-Food Canada, Saskatoon Research Centre, 107 Science Place, Saskatoon, SK S7N 0X2, Canada. gruberm@agr.gc.ca Transformation with the Arabidopsis bHLH gene 35S:GLABRA3 (GL3) produced novel B. napus plants with an extremely dense coverage of trichomes on seedling tissues (stems and young leaves). In contrast, trichomes were strongly induced in seedling stems and moderately induced in leaves of a hairy, purple phenotype transformed with a 2.2 kb allele of the maize anthocyanin regulator LEAF COLOUR (Lc), but only weakly induced by BOOSTER (B-Peru), the maize Lc 2.4 kb allele, or the Arabidopsis trichome MYB gene GLABRA1 (GL1). B. napus plants containing only the GL3 transgene had a greater proportion of trichomes on the adaxial leaf surface, whereas all other plant types had a greater proportion on the abaxial surface. Progeny of crosses between GL3+ and GL1+ plants resulted in trichome densities intermediate between a single-insertion GL3+ plant and a double-insertion GL3+ plant. None of the transformations stimulated trichomes on Brassica cotyledons or on non-seedling tissues. A small portion of bHLH gene-induced trichomes had a swollen terminal structure. The results suggest that trichome development in B. napus may be regulated differently from Arabidopsis. They also imply that insertion of GL3 into Brassica species under a tissue-specific promoter has strong potential for developing insect-resistant crop plants. Publication Types: Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. PMID: 16649106 [PubMed - indexed for MEDLINE] 730: J Am Diet Assoc. 2006 May;106(5):719-27. Use of a risk communication model to evaluate dietetics professionals' viewpoints on genetically engineered foods and crops. Roberts KS, Struble MB, McCullum-Gomez C, Wilkins JL. Hunterdon Medical Center, Flemington, NJ, USA. KRoberts@cse.edu The complex issues surrounding the application of genetic engineering to food and agriculture have generated a contentious debate among diverse interest groups. One pervasive dimension in the resultant discourse is the varying perceptions of the risks and benefits of genetically engineered foods and crops. In the risk communication model, technical information is evaluated within the context of an individual's values and perceptions. The purpose of this study was to explore how dietetics professionals respond to a complex set of interrelated issues associated with genetically engineered foods and crops and to identify what varying viewpoints may exist. Participants were asked to sort a total of 48 statements distributed across eight issue areas according to level of agreement and disagreement. Using Q methodology, a total of 256 sortings were analyzed using the centroid method and varimax rotation in factor analysis. Three distinct viewpoints emerged: Precautionary (R(2)=43%), Discerning Supporter (R(2)=11%), and Promoting (R(2)=5%). Across all viewpoints, respondents agreed that dietetics professionals should employ critical thinking skills to communicate the social, economic, environmental, ethical, and technical aspects of genetically engineered foods and crops. The findings have implications for how dietetics professionals can foster an open interchange of information among diverse groups. Publication Types: Research Support, Non-U.S. Gov't PMID: 16647331 [PubMed - indexed for MEDLINE] 731: J Biotechnol. 2006 Oct 1;125(4):447-61. Epub 2006 Apr 27. The effect of polysaccharide-degrading wine yeast transformants on the efficiency of wine processing and wine flavour. Louw C, La Grange D, Pretorius IS, van Rensburg P. Institute for Wine Biotechnology, Department of Viticulture and Oenology, Stellenbosch University, Matieland (Stellenbosch), ZA 7602, South Africa. Commercial polysaccharase preparations are applied to winemaking to improve wine processing and quality. Expression of polysaccharase-encoding genes in Saccharomyces cerevisiae allows for the recombinant strains to degrade polysaccharides that traditional commercial yeast strains cannot. In this study, we constructed recombinant wine yeast strains that were able to degrade the problem-causing grape polysaccharides, glucan and xylan, by separately integrating the Trichoderma reesei XYN2 xylanase gene construct and the Butyrivibrio fibrisolvens END1 glucanase gene cassette into the genome of the commercial wine yeast strain S. cerevisiae VIN13. These genes were also combined in S. cerevisiae VIN13 under the control of different promoters. The strains that were constructed were compared under winemaking conditions with each other and with a recombinant wine yeast strain expressing the endo-beta-1,4-glucanase gene cassette (END1) from B. fibrisolvens and the endo-beta-1,4-xylanase gene cassette (XYN4) from Aspergillus niger, a recombinant strain expressing the pectate lyase gene cassette (PEL5) from Erwinia chrysanthemi and the polygalacturonase-encoding gene cassette (PEH1) from Erwinia carotovora. Wine was made with the recombinant strains using different grape cultivars. Fermentations with the recombinant VIN13 strains resulted in significant increases in free-flow wine when Ruby Cabernet must was fermented. After 6 months of bottle ageing significant differences in colour intensity and colour stability could be detected in Pinot Noir and Ruby Cabernet wines fermented with different recombinant strains. After this period the volatile composition of Muscat d'Alexandria, Ruby Cabernet and Pinot Noir wines fermented with different recombinant strains also showed significant differences. The Pinot Noir wines were also sensorial evaluated and the tasting panel preferred the wines fermented with the recombinant strains. Publication Types: Evaluation Studies PMID: 16644051 [PubMed - indexed for MEDLINE] 732: Appl Microbiol Biotechnol. 2006 Aug;71(5):598-607. Epub 2006 Apr 26. Genetically modified crops: success, safety assessment, and public concern. Singh OV, Ghai S, Paul D, Jain RK. Department of Pediatrics, The John Hopkins School of Medicine, Baltimore, MD 21287, USA. osingh1@jhmi.edu With the emergence of transgenic technologies, new ways to improve the agronomic performance of crops for food, feed, and processing applications have been devised. In addition, ability to express foreign genes using transgenic technologies has opened up options for producing large quantities of commercially important industrial or pharmaceutical products in plants. Despite this high adoption rate and future promises, there is a multitude of concerns about the impact of genetically modified (GM) crops on the environment. Potential contamination of the environment and food chains has prompted detailed consideration of how such crops and the molecules that they produce can be effectively isolated and contained. One of the reasonable steps after creating a transgenic plant is to evaluate its potential benefits and risks to the environment and these should be compared to those generated by traditional agricultural practices. The precautionary approach in risk management of GM plants may make it necessary to monitor significant wild and weed populations that might be affected by transgene escape. Effective risk assessment and monitoring mechanisms are the basic prerequisites of any legal framework to adequately address the risks and watch out for new risks. Several agencies in different countries monitor the release of GM organisms or frame guidelines for the appropriate application of recombinant organisms in agro-industries so as to assure the safe use of recombinant organisms and to achieve sound overall development. We feel that it is important to establish an internationally harmonized framework for the safe handling of recombinant DNA organisms within a few years. Publication Types: Review PMID: 16639559 [PubMed - indexed for MEDLINE] 733: J Agric Food Chem. 2006 May 3;54(9):3173-80. Need for an "integrated safety assessment" of GMOs, linking food safety and environmental considerations. Haslberger AG. Vienna Ecology Center, Department for Nutritional Sciences, University of Vienna, Althanstrasse 2, A-1090 Vienna, Austria. alexander.haslberger@univie.ac.at Evidence for substantial environmental influences on health and food safety comes from work with environmental health indicators which show that agroenvironmental practices have direct and indirect effects on human health, concluding that "the quality of the environment influences the quality and safety of foods" [Fennema, O. Environ. Health Perspect. 1990, 86, 229-232). In the field of genetically modified organisms (GMOs), Codex principles have been established for the assessment of GM food safety and the Cartagena Protocol on Biosafety outlines international principles for an environmental assessment of living modified organisms. Both concepts also contain starting points for an assessment of health/food safety effects of GMOs in cases when the environment is involved in the chain of events that could lead to hazards. The environment can act as a route of unintentional entry of GMOs into the food supply, such as in the case of gene flow via pollen or seeds from GM crops, but the environment can also be involved in changes of GMO-induced agricultural practices with relevance for health/food safety. Examples for this include potential regional changes of pesticide uses and reduction in pesticide poisonings resulting from the use of Bt crops or influences on immune responses via cross-reactivity. Clearly, modern methods of biotechnology in breeding are involved in the reasons behind the rapid reduction of local varieties in agrodiversity, which constitute an identified hazard for food safety and food security. The health/food safety assessment of GM foods in cases when the environment is involved needs to be informed by data from environmental assessment. Such data might be especially important for hazard identification and exposure assessment. International organizations working in these areas will very likely be needed to initiate and enable cooperation between those institutions responsible for the different assessments, as well as for exchange and analysis of information. An integrated assessment might help to focus and save capacities in highly technical areas such as molecular characterization or profiling, which are often necessary for both assessments. In the area of establishing international standards for traded foods, such as for the newly created Standards in Trade and Development Facility (STDF), an integrated assessment might help in the consideration of important environmental aspects involved in health and food safety. Furthermore, an established integrated view on GMOs may create greater consumer confidence in the technology. Publication Types: Research Support, Non-U.S. Gov't PMID: 16637668 [PubMed - indexed for MEDLINE] 734: Biosci Biotechnol Biochem. 2006 Apr;70(4):821-7. Quantification of genetically modified soybeans using a combination of a capillary-type real-time PCR system and a plasmid reference standard. Toyota A, Akiyama H, Sugimura M, Watanabe T, Kikuchi H, Kanamori H, Hino A, Esaka M, Maitani T. Hiroshima Prefectural Institute of Public Health and Environment, Minami-ku, Hiroshima, Japan. Because the labeling of grains and feed- and foodstuffs is mandatory if the genetically modified organism (GMO) content exceeds a certain level of approved genetically modified varieties in many countries, there is a need for a rapid and useful method of GMO quantification in food samples. In this study, a rapid detection system was developed for Roundup Ready Soybean (RRS) quantification using a combination of a capillary-type real-time PCR system, a LightCycler real-time PCR system, and plasmid DNA as the reference standard. In addition, we showed for the first time that the plasmid and genomic DNA should be similar in the established detection system because the PCR efficiencies of using plasmid DNA and using genomic DNA were not significantly different. The conversion factor (Cf) to calculate RRS content (%) was further determined from the average value analyzed in three laboratories. The accuracy and reproducibility of this system for RRS quantification at a level of 5.0% were within a range from 4.46 to 5.07% for RRS content and within a range from 2.0% to 7.0% for the relative standard deviation (RSD) value, respectively. This system rapidly monitored the labeling system and had allowable levels of accuracy and precision. Publication Types: Research Support, Non-U.S. Gov't PMID: 16636447 [PubMed - indexed for MEDLINE] 735: Environ Biosafety Res. 2005 Jul-Sep;4(3):179-88. Potential adoption and management of insect-resistant potato in Peru, and implications for genetically engineered potato. Buijs J, Martinet M, de Mendiburu F, Ghislain M. International Potato Center, Apartado 1558, Lima 12, Peru. jasper_buijs25@yahoo.com This paper analyzes some important issues surrounding possible deployment of genetically engineered (GE) insect-resistant potato in Peru, based on a large farmer survey held in Peru in 2003. We found that the formal seed system plays a limited role compared with the informal seed system, especially for smallholder farmers. Although 97% of smallholder farmers would buy seed of an insect-resistant variety, a majority would buy it only once every 2 to 4 years. Survey data show that farmers would be willing to pay a premium of 50% on seed cost for insect resistant varieties. Paying price premiums of 25% to 50%, farmers would still increase their net income, assuming insect resistance is high and pesticide use will be strongly reduced. Of all farmers, 55% indicated preference for insect-resistant potato in varieties other than their current varieties. The survey indicates that smallholder farmers are interested to experiment with new varieties and have a positive perception of improved varieties. Based on these findings, and considering the difficulties implementing existing biosafety regulatory systems such as those in place in the U.S. and E.U., we propose to develop a variety-based segregation system to separate GE from conventionally bred potatoes. In such a system, which would embrace the spread of GE potatoes through informal seed systems, only a limited number of sterile varieties would be introduced that are easily distinguishable from conventional varieties. Publication Types: Comparative Study Research Support, Non-U.S. Gov't PMID: 16634223 [PubMed - indexed for MEDLINE] 736: Environ Biosafety Res. 2005 Jul-Sep;4(3):141-66. Understanding the potential impact of transgenic crops in traditional agriculture: maize farmers' perspectives in Cuba, Guatemala and Mexico. Soleri D, Cleveland DA, Aragón F, Fuentes MR, Ríos H, Sweeney SH. Environmental Studies Program and Geography Department, University of California, Santa Barbara, 2302 Girvetz Hall, Santa Barbara, CA 93106, USA. soleri@es.ucsb.edu Genetically engineered transgenic crop varieties (TGVs) have spread rapidly in the last 10 years, increasingly to traditionally-based agricultural systems (TBAS) of the Third World both as seed and food. Proponents claim they are key to reducing hunger and negative environmental impacts of agriculture. Opponents claim they will have the opposite effect. The risk management process (RMP) is the primary way in which TGVs are regulated in the US (and many other industrial countries), and proponents claim that the findings of that process in the US and its regulatory consequences should be extended to TBAS. However, TBAS differ in important ways from industrial agriculture, so TGVs could have different effects in TBAS, and farmers there may evaluate risks and benefits differently. To evaluate some potential impacts of TGVs in TBAS we used the RMP as a framework for the case of Bt maize in Mesoamerica and Cuba. We interviewed 334 farmers in Cuba, Guatemala and Mexico about farming practices, evaluations of potential harm via hypothetical scenarios, and ranking of maize types. Results suggest high potential for transgene flow via seed, grain and pollen; differences in effects of this exposure in TBAS compared with industrial agriculture; farmers see some potential consequences as harmful. Perceptions of harm differ among farmers in ways determined by their farming systems, and are different from those commonly assumed in industrial systems. An RMP including participation of farmers and characteristics of TBAS critical for their functioning is necessary to ensure that investments in agricultural technologies will improve, not compromise these agricultural systems. Publication Types: Comparative Study Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. PMID: 16634221 [PubMed - indexed for MEDLINE] 737: Law Hum Genome Rev. 2005 Jul-Dec;(23):217-21. Report on genetically modified organisms in agriculture and food. [Article in English, Spanish] Ethics Advisory Committe on Scientific and Technological Research. Publication Types: Practice Guideline PMID: 16628882 [PubMed - indexed for MEDLINE] 738: Pest Manag Sci. 2006 Jun;62(6):558-64. Metabolism of [14C]-2,4-dichlorophenol in edible plants. Laurent F, Debrauwer L, Pascal-Lorber S. INRA, UMR Xénobiotiques, 180 Ch. de Tournefeuille, BP3, F-31931 Toulouse Cedex 9, France. flaurent@toulouse.inra.fr Several 2,4-dichlorophenoxyacetic acid (2,4-D)-sensitive plants have been modified by genetic engineering with tfdA gene to acquire 2,4-D tolerance. The expression product of this gene degrades 2,4-D to 2,4-dichlorophenol (DCP), which is less phytotoxic but could cause a problem of food safety. After a comparison of 2,4-D and DCP metabolism in transgenic 2,4-D-tolerant and wild cotton (Gossypium hirsutum L.), a direct study of DCP metabolism in edible plants was performed. After petiolar uptake of a [U-phenyl-(14)C]-DCP solution followed by a 48 h water chase, aqueous extracts were analysed by high-performance liquid chromatography. Metabolites were thereafter isolated and their structural identities were determined by enzymatic and chemical hydrolyses and mass spectrometry analyses. The metabolic fate of DCP was equivalent to 2,4-D metabolism in transgenic 2,4-D-tolerant cotton. In addition, DCP metabolism was similar in transgenic and wild cotton. The major terminal metabolites were DCP-saccharide conjugates in all species, essentially DCP-(6-O-malonyl)-glucoside or its precursor DCP-glucose. The significance of this metabolic pathway with regard to food safety is discussed. Copyright (c) 2006 Society of Chemical Industry Publication Types: Comparative Study PMID: 16628540 [PubMed - indexed for MEDLINE] 739: Sci Cult (Lond). 2005 Dec;14(4):393-410. Asilomar's legacy in Aotearoa New Zealand. Rogers-Hayden T. Centre for Environmental Risk, Environmental Sciences, University of East Anglis, Norwich NR4 7TJ, UK. T.Rogers-Hayden@uea.ac.uk Publication Types: Historical Article Research Support, Non-U.S. Gov't PMID: 16622954 [PubMed - indexed for MEDLINE] 740: Sci Cult (Lond). 2005 Dec;14(4):373-92. Genetic engineering regulation in Australia: an 'archaeology' of expertise and power. Hindmarsh R. Australian School of Environmental Sciences, Griffith University, Nathan, Brisbane, 4111, Australia. r.hindmarsh@griffith.edu.au Publication Types: Research Support, Non-U.S. Gov't PMID: 16622953 [PubMed - indexed for MEDLINE] 741: Shokuhin Eiseigaku Zasshi. 2006 Feb;47(1):15-27. [Laboratory-performance study of quantitative PCR methods to analyze an approved genetically modified maize (Mon810 Line)] [Article in Japanese] Watanabe T, Kasama K, Kikuchi H, Suzuki T, Tokishita S, Sakata K, Matsuki A, Hino A, Akiyama H, Maitani T. National Institute of Health Sciences: 1-18-1, Kamiyoga, Setagaya-ku, Tokyo 158-8501, Japan. A laboratory-performance study was carried out to investigate factors affecting the reliability of the quantitative PCR method to analyze an approved genetically modified (GM) maize (Mon810 line). Test maize powdered samples were prepared as blind samples containing a high (assigned value; 5.45%) or low (assigned value; 0.35%) concentration of the Mon810 line. After confirmation of their homogeneity, they were provided to 27 laboratories participating in the collaborative study. The data were collected from all laboratories and statistically analyzed. Two laboratories, which used a Roche LightCycler (LC), reported significantly high test values. A further examination showed that the LC method is greatly affected by the equipment itself or PCR reagents, resulting in poor repeatability. On the other hand, some laboratories, which used ABI quantitative PCR equipment, reported erroneous test values. In these laboratories, the errors appeared to have been due to inadequate quality and/or yield of DNA. To identify factors affecting the test values, analysis of the measured values for the taxon-specific gene will be useful. Furthermore, the modified silica-gel membrane DNA extraction method made it possible to extract the required amounts of DNA more easily and in a shorter time than before. Publication Types: English Abstract Research Support, Non-U.S. Gov't PMID: 16619852 [PubMed - indexed for MEDLINE] 742: Shokuhin Eiseigaku Zasshi. 2006 Feb;47(1):9-14. Investigation of false-positive reactions for CBH351 maize in screening PCR analysis. Monma K, Moriuchi R, Sagi N, Ichikawa H, Satoh K, Tobe T, Kamata K. Tokyo Metropolitan Institute of Public Health, 3-24-1, Hyakunin-cho, Shinjuku-ku, Tokyo 169-0073, Japan. Examination for CBH351 maize was conducted by the qualitative polymerase chain reaction (PCR) method in maize grain and maize processed foods obtained in the Tokyo area. The numbers of samples possibly positive in the screening test were 7 of 22 (31.8%) for maize grain samples, 4 of 14 (28.6%) for semi-processed foods, 11 of 30 (36.7%) for canned products, 3 of 30 (10.0%) for maize snacks, 3 of 4 (75%) for tacos and 1 of 3 (33.3%) for tortillas. However, CBH351 maize was not detected in the confirmation test. Therefore, the results of the screening test were false-positive. Since the reaction might have been caused by the base sequences of the 3'-end of primers CaM03-5' and CBH02-3' used in the screening test, a new primer pair was designed. The PCR products obtained with the new primer pair TMC2-5'--TMS2-3' were specific for CBH351 and were not obtained with barley, wheat, rice, RRS, Bt11, or Event176. Thus, the new primer pair shows high specificity. CBH351 maize was detected from samples containing at least 0.05% CBH 351 maize DNA by using this primer pair. PMID: 16619851 [PubMed - indexed for MEDLINE] 743: Food Nutr Bull. 2002 Dec;23(4):378-81. Biotechnology-derived nutritious foods for developing countries: needs, opportunities, and barriers: discussion summary. [No authors listed] Improvements in diet diversification and quality can be facilitated by greater cooperation between the agricultural and the nutrition communities, according to an expert panel that met in early 2002. Encouraged to think innovatively, the panelists agreed that modern technology offers the potential to increase the amount and nutritional content of the food supply in developing countries, especially if the enhancements are made to the highest-yielding indigenous staple crops and if a total food-systems approach is taken. All types of interventions should be evaluated for their cost-effectiveness in preventing nutritional deficiencies in the developing world and for their sustainability. Publication Types: Congresses PMID: 16619745 [PubMed - indexed for MEDLINE] 744: Food Nutr Bull. 2002 Dec;23(4):364-6. The potential for biotechnology to improve the nutritional value of cassava. Fauquet CM, Taylor N. International Laboratory for Tropical Agricultural Biotechnology, Donald Danforth Plant Science Center, in St. Louis, Missouri, USA. Cassava, a starch-rich plant that has poor protein content and usually poor vitamin content, feeds about 600 million people each day. When cereals can no longer be grown because of soil fertility problems, it is often still possible to grow cassava. It is the third most important source of dietary calories in the tropics, and reliance on the crop is especially high in West and Central Africa. The International Laboratory for Tropical Agricultural Biotechnology is promoting research to improve cassava productivity and is a leader in developing genetic engineering to improve the quantitative and qualitative traits of this essential food crop. PMID: 16619742 [PubMed - indexed for MEDLINE] 745: Food Nutr Bull. 2002 Dec;23(4):360-3. Food biotechnology and nutrition in Africa: a case for Kenya. Ngichabe CK. Kenya Agricultural Research Institute in Nairobi. Household food consumption surveys indicate that the diet in Kenya is ill balanced and that many families cannot afford nutrient-rich foods such as meat and fruits. In this regard, rural populations-the majority of the Kenyan population-are much worse off than urban populations. Agriculture, the most important sector in the Kenyan economy, contributes 27% of the gross domestic product and generates 65% of the country's export earnings. Food-enhancing biotechnologies thus could increase national food yields and fill nutrition gaps by contributing to household and national food security and poverty reduction in Kenya. To overcome barriers to adopting biotechnology to improve food crops in Kenya and elsewhere in Africa, policy makers must create a receptive environment for, increase public understanding of, and stimulate investment in the new technology. PMID: 16619741 [PubMed - indexed for MEDLINE] 746: Food Nutr Bull. 2002 Dec;23(4):358-9. Can biotechnology help meet the nutrition challenge in sub-Saharan Africa? Tagwireyi J. Ministry of Finance and Economic Development in Harare, Zimbabwe. The successful efforts in the 1980s to redress nutrition problems in sub-Saharan Africa are being eroded. Countries in eastern and southern Africa are now facing serious food shortages because of recurrent droughts, floods, civil wars, and the concomitant growing poverty. The potential for biotechnology to alleviate hunger holds promise if the new technology can be adapted to the prevailing sociocultural context in Africa. Agronomists and biotechnologists need to work together to ensure that the biotechnology agenda for Africa is responsive to the food and nutrition needs of its people. PMID: 16619740 [PubMed - indexed for MEDLINE] 747: Food Nutr Bull. 2002 Dec;23(4):354-7. The promise of biotechnology in addressing current nutritional problems in developing countries. Khush GS. The author is affiliated with the International Rice Research Institute, Los Baños, Laguna, Philippines. To meet the nutritional needs of a rapidly growing world population, which is likely to reach 8 billion by 2030, 50% more food grains with higher and more stable yields must be produced. Biofortification is considered the most effective way to increase micronutrient intakes. It is low cost and sustainable and does not require a change in eating habits or impose recurring costs. A research project to improve the iron and zinc content of rice was initiated at the International Rice Research Institute in 1992. Several experimental lines of rice with increased iron and zinc content have been produced. In another experiment rices with beta-carotene have been produced. Other experimental efforts aim at raising the micronutrient content in wheat, maize, cassava, sweet potatoes, and beans. Maize with improved amino acid balance is being grown in several African countries. PMID: 16619739 [PubMed - indexed for MEDLINE] 748: Food Nutr Bull. 2002 Dec;23(4):351-3. Three criteria for establishing the usefulness of biotechnology for reducing micronutrient malnutrition. Bouis HE. International Food Policy Research Institute, 2033 K St NW, Washington, DC 20006, USA. The fundamental reason that plant breeding using either conventional breeding or biotechnology is so cost-effective is that the benefits of a one-time investment at a central research location can be multiplied over time across nations all over the world. Supplementation and fortification incur the same recurrent costs year after year in country after country. However, each intervention has its own comparative advantages, such that a combination of several interventions is required to substantially reduce micronutrient malnutrition. Improving the density of trace minerals in plants also reduces input requirements and raises crop yields. A simulation model for India and Bangladesh demonstrated that $42 million invested in conventional breeding in developing and planting iron- and zinc-dense varieties of rice and wheat on only 10% of the acreage used for these crops would return $4.9 billion in improved nutrition (including a total of 44 million prevented cases of anemia over 10 years) and higher agricultural productivity. PMID: 16619738 [PubMed - indexed for MEDLINE] 749: Sci Cult (Lond). 2005 Dec;14(4):355-72. Genetically modified survival: red and green biotechnology in Israel. Prainsack B, Firestine O. Department of Political Science, University of Vienna, Universitaetsstrasse 7, A-1010 Wein, Austria. barbara.prainsack@univie.ac.at PMID: 16619470 [PubMed - indexed for MEDLINE] 750: Sci Cult (Lond). 2005 Dec;14(4):339-53. The long and winding road from Asilomar to Brussels: science, politics and the public in biotechnology and regulation. Abels G. Institute for Science and Technology Studies, Bielefeld University, P.O. Box 10 01 31, D-33501 Bielefeld, Germany. abels@iwt.uni-bielefeld.de Publication Types: Historical Article PMID: 16619469 [PubMed - indexed for MEDLINE] 751: Sci Cult (Lond). 2005 Dec;14(4):309-23. From Asilomar to industrial biotechnology: risks, reductionism and regulation. Krimsky S. Department of Urban & Environmental Policy & Planning, Tufts University, Medford, MA 02155, USA. sheldon.krimsky@tufts.edu Publication Types: Historical Article PMID: 16619467 [PubMed - indexed for MEDLINE] 752: Food Nutr Bull. 2005 Dec;26(4):403-52. Proceedings of the Symposium and Workshop on Biotechnology Derived Nutritious Foods: Challenges and Opportunities in Asia. February 29-March 1, 2004, Bali, Indonesia. [No authors listed] Publication Types: Congresses Overall PMID: 16619434 [PubMed - indexed for MEDLINE] 753: Biotechnol Lett. 2006 Mar;28(5):321-5. The occurrence of antibiotic resistance genes in Taq polymerases and a decontamination method applied to the detection of genetically modified crops. Perron A, Raymond P, Simard R. St-Hyacinthe Laboratory, Canadian Food Inspection Agency, Casavant Blvd West, J2S 8E3 3400, St-Hyacinthe, Quebec, Canada. Different antibiotic resistance (AR) genes, such as Bla, Tet and NPTII, contaminate commercially available Taq polymerases. The specificity of the AR gene PCR can be increased when using a restriction enzyme-based decontamination of polymerase. The elimination of Taq polymerase contamination allows the use of PCR tests to screen seeds (corn) and processed food for the presence of genetically modified organisms (GMO) based on the detection of AR genes. Without a decontamination procedure for AR genes, PCR screening tests should be interpreted with caution. Publication Types: Research Support, Non-U.S. Gov't PMID: 16614919 [PubMed - indexed for MEDLINE] 754: J Agric Food Chem. 2006 Apr 19;54(8):2799-809. Coherence between legal requirements and approaches for detection of genetically modified organisms (GMOs) and their derived products. Holst-Jensen A, De Loose M, Van den Eede G. National Veterinary Institute, Ullevaalsveien 68, P.O. Box 8156 Dep., 0033 Oslo, Norway. arne.holst-jensen@vetinst.no Analytical methods for the qualitative and quantitative detection of genetically modified (GM) products may serve multiple purposes. Legal requirements differ among jurisdictions, ranging from no requirements to mandatory use of event-specific quantitation and implementation of production chain traceability. Although efforts have been taken to harmonize the analytical methodology at national, regional, and international levels, no normative international standards have yet been established. Lack of coherence between analytical methodologies and their applicabilities, on the one hand, and legislation, on the other hand, is a major problem. Here, key points where coherence is lacking are discussed. These include the definition of units of measurements, expression of GM material quantities, terminology, and inconsistent legal status of products derived from related but slightly different transformation routes. Finally, recommendations to improve the coherence are brought forward, including guidance to stakeholders for prediction of product-specific GM material quantities from gene ratios in the originating seed. Publication Types: Research Support, Non-U.S. Gov't PMID: 16608192 [PubMed - indexed for MEDLINE] 755: Transgenic Res. 2006 Apr;15(2):131-7. Metabolic engineering of plants to produce very long-chain polyunsaturated fatty acids. Truksa M, Wu G, Vrinten P, Qiu X. Bioriginal Food & Science Corporation, Saskatoon, Saskatchewan, Canada. Very long-chain polyunsaturated fatty acids (VLCPUFAs) are essential for human health and well-being. However, the current sources of these valuable compounds are limited and may not be sustainable in the long term. Recently, considerable progress has been made in identifying genes involved in the biosynthesis of VLCPUFAs. The co-expression of these genes in model systems such as plant embryos or yeast provided many valuable insights into the mechanisms of VLCPUFA synthesis. The recent successful reconstitution of pathways leading to the synthesis of arachidonic acid, eicosapentaenoic acid and finally docosahexaenoic acid in oil-seed plants indicates the feasibility of using transgenic crops as alternative sources of VLCPUFAs. The various approaches used to attain these results and the specific constraints associated with each approach are discussed. Publication Types: Review PMID: 16604455 [PubMed - indexed for MEDLINE] 756: Ann Allergy Asthma Immunol. 2006 Mar;96(3 Suppl 2):S1-68. Food allergy: a practice parameter. American College of Allergy, Asthma, & Immunology. Publication Types: Practice Guideline PMID: 16597066 [PubMed - indexed for MEDLINE] 757: Exp Appl Acarol. 2006;38(2-3):125-39. Assessing the effects of Bt Maize on the predatory mite Neoseiulus cucumeris. Obrist LB, Klein H, Dutton A, Bigler F. Agroscope FAL Reckenholz, Swiss Federal Research Station for Agroecology and Agriculture, Reckenholzstr. 191, 8046, Zurich, Switzerland. The investigation of Neoseiulus cucumeris in the context of the ecological risk assessment of insect resistant transgenic plants is of particular interest as this omnivorous predatory mite species is commercially available and considered important for biological control. In a multitrophic feeding experiment we assessed the impact of Bt maize on the performance of N. cucumeris when offered spider mites (Tetranychus urticae) reared on Bt (Bt11, Syngenta) or non-Bt maize (near isogenic line) and Bt or non-Bt maize pollen as a food source. Various parameters including mortality, development time, oviposition rate were measured. Spider mites were used as a prey for N. cucumeris, since these herbivores are known to contain similar levels of Cry1Ab toxin, when reared on Bt maize, as those found in the transgenic leaf material. In contrast, toxin levels in pollen of this transgenic cultivar are very low. No differences in any of the parameters were found when N. cucumeris was fed with spider mites reared on Bt and non-Bt maize. Pollen was shown to be a less suitable food source for this predator as compared to spider mites. Moreover, subtle effects on female N. cucumeris (9% longer development time and 17% reduced fecundity) were measured when fed with pollen originating from Bt maize as compared to non-Bt maize pollen. Our findings indicate that the predatory mite N. cucumeris is not sensitive to the Cry1Ab toxin as no effects could be detected when offered Bt-containing spider mites, and that the effects found when fed with Bt maize pollen can be assigned to differences in nutritional quality of Bt and non-Bt maize pollen. The significance of these findings is discussed with regard to the ecological relevance for risk assessment of transgenic plants. Publication Types: Research Support, Non-U.S. Gov't PMID: 16596347 [PubMed - indexed for MEDLINE] 758: J Exp Bot. 2006;57(9):1899-908. Epub 2006 Apr 4. Oxalate accumulation and regulation is independent of glycolate oxidase in rice leaves. Xu HW, Ji XM, He ZH, Shi WP, Zhu GH, Niu JK, Li BS, Peng XX. Laboratory of Molecular Plant Physiology, College of Life Sciences, South China Agricultural University, Guangzhou, PR China. xpeng@scau.edu.cn Cellular oxalate, widely distributed in many plants, is implicated to play important roles in various functions and is also known to affect food qualities adversely in fruits and vegetables. How oxalate is regulated in plants is currently not well understood. Glycolate oxidase (GLO) has long been considered as an important player in oxalate accumulation in plants. To gain further insight into the biochemical and molecular mechanisms, the possible roles of GLO in the process were studied. Drastically different levels of oxalate could be achieved by treating rice with various nitrogen forms (nitrate versus ammonium). While nitrate stimulated oxalate accumulation, ammonium reduced its level. Such treatments resulted in similar pattern changes for some other related organic acids, such as glycolate, oxaloacetate, and malate. By feeding plants with exogenous glycolate it was possible almost completely to restore the ammonium-decreased oxalate level. Under the two treatments few differences were observed for GLO mRNA levels, protein levels, and in vitro activities. Both K(m) for glycolate/glyoxylate and K(i) for oxalate remained almost the same for GLO purified from either nitrate- or ammonium-fed leaves. A further in vivo study, with transgenic plants carrying an estradiol-inducible GLO antisense gene, showed that, while the estradiol-induced antisense expression remarkably reduced both GLO protein levels and activities, oxalate levels were not significantly altered in the estradiol-treated transgenic plants. Taken together, it is suggested that oxalate accumulation and regulation is independent of GLO in rice leaves. Publication Types: Research Support, Non-U.S. Gov't PMID: 16595582 [PubMed - indexed for MEDLINE] 759: Appl Microbiol Biotechnol. 2006 May;70(6):642-50. Epub 2006 Apr 4. Molecular plant breeding: achievements in green biotechnology and future perspectives. Wenzel G. Plant Breeding, Center for Life and Food Sciences, Technische Universität München, Freising, Germany. gwenzel@wzw.tum.de Since one decade ago, transgenic crop plants are globally grown; in 2004, it was estimated to cover a total of 81 Mio ha in 17 countries. At present, four plant species (soybean, maize, cotton and rapeseed) dominate with two traits (herbicide tolerance and insect resistance). The traits on which research concentrates and the constructs which might come next onto the market are outlined. The procedure on how to clone such genes of interest, e.g. via map-based cloning, and some other helpful approaches of green biotechnology, like high throughput techniques and functional markers, are summarised, and a rough calculation about the market value of transgenic crops in US dollars is quoted. Publication Types: Review PMID: 16586104 [PubMed - indexed for MEDLINE] 760: Science. 2006 Mar 31;311(5769):1940-2. Comment in: Science. 2006 Mar 31;311(5769):1872-3. Cellulose synthase-like CslF genes mediate the synthesis of cell wall (1,3;1,4)-beta-D-glucans. Burton RA, Wilson SM, Hrmova M, Harvey AJ, Shirley NJ, Medhurst A, Stone BA, Newbigin EJ, Bacic A, Fincher GB. Australian Centre for Plant Functional Genomics; School of Agriculture, Food, and Wine; University of Adelaide, Waite Campus, Glen Osmond, SA 5064, Australia. A characteristic feature of grasses and commercially important cereals is the presence of (1,3;1,4)-beta-d-glucans in their cell walls. We have used comparative genomics to link a major quantitative trait locus for (1,3;1,4)-beta-d-glucan content in barley grain to a cluster of cellulose synthase-like CslF genes in rice. After insertion of rice CslF genes into Arabidopsis, we detected (1,3;1,4)-beta-d-glucan in walls of transgenic plants using specific monoclonal antibodies and enzymatic analysis. Because wild-type Arabidopsis does not contain CslF genes or have (1,3;1,4)-beta-d-glucans in its walls, these experiments provide direct, gain-of-function evidence for the participation of rice CslF genes in (1,3;1,4)-beta-d-glucan biosynthesis. Publication Types: Research Support, Non-U.S. Gov't PMID: 16574868 [PubMed - indexed for MEDLINE] 761: Risk Anal. 2006 Apr;26(2):455-70. Loss of agro-biodiversity, uncertainty, and perceived control: a comparative risk perception study in Austria and China. Schmidt MR, Wei W. University of Vienna, Institute of Risk Research, Vienna, Austria. ms@irf.univie.ac.at The biogeographical centers of origin of important food crops-called Vavilov centers-are considered to be crucial sources of genetic diversity for present and future crop-breeding programs and thus for human food safety worldwide. Global environmental change and more intensified modes of crop production may cause genetic erosion (loss of traditional crop varieties and loss of crop wild relatives), especially in Vavilov centers. The present study focused on how the risk of genetic erosion (or loss of agro-biodiversity) is perceived in comparison to 16 other risk topics by experts and lay people in Austria and China. The most striking result was that genetic erosion was perceived to be an exceptionally unknown and uncertain risk topic, given that only genetically modified organisms (GMOs) were perceived as being even more uncertain. As a consequence of the high uncertainty, the idea of applying the precautionary principle to further prevent genetic erosion is discussed. An unprecedented finding-one that differs from Austrian participants-is that the Chinese have a higher perceived control over all risk topics. The increased perception of controllability in China is discussed in light of the theory of reflexive modernization. This theory strives to explain the increased critical attitude in Western countries such as Austria toward scientific innovations and toward the idea that everything can be calculated and mastered at will. By revealing different notions of risk perception, this research also provides additional scientific input to risk communication efforts for public education. Publication Types: Comparative Study Research Support, Non-U.S. Gov't PMID: 16573633 [PubMed - indexed for MEDLINE] 762: Trends Biotechnol. 2006 May;24(5):206-11. Epub 2006 Mar 29. Global trends in plant transgenic science and technology (1973-2003). Vain P. John Innes Centre, Crop Genetics Department, Norwich Research Park, UK, NR4 7UH. philippe.vain@bbsrc.ac.uk Transgenic science and technology are fundamental to state-of-the-art plant molecular genetics and GM crop improvement. Monitoring the scale and growth of this area of science is important to scientists, national and international research organizations, funding bodies, policy makers and, because of the GM debate, to society as a whole. Literature statistics covering the past 30 years reveal a dramatic increase in plant transgenic science in Asia during the past decade, a sustained expansion in North America and, recently, a slow down in the rest of the world. With the exception of the output of China and India, publications focusing on the development of transgenic technology have been slowing down, worldwide, since the early mid-1990s, a trend that contrasts with the increase in GM crop-related studies. PMID: 16569453 [PubMed - indexed for MEDLINE] 763: Curr Allergy Asthma Rep. 2006 Mar;6(2):153-9. Novel foods to treat food allergy and gastrointestinal infection. Perr HA. Evolving Foods and Children's Health, Division of Pediatric Gastroenterology, Hepatology, and Nutrition, California Pacific Medical Center, Box 7999, San Francisco, 94120, USA. hperr@itsa.ucsf.edu The gastrointestinal tract communicates directly with the external environment. Necessary nutrients must be absorbed and commensal bacteria tolerated, and foreign proteins, antigens, and pathogens must be simultaneously excluded or destroyed. Immaturity or disruption of the mucosal immune defenses increases vulnerability to food allergy, intolerance, and infectious disease. Diseases resulting from ingested foreign proteins and organisms are increasing and cause morbidity and mortality worldwide. There is no specific treatment for food allergy other than avoidance. Vaccination for infectious disease is limited by the cost and logistics of distribution and administration, particularly in developing countries. Novel strategies are being explored to modulate the gut mucosal immune system by altering protein expression in food. Crops are being developed to remove deleterious allergens to prevent immunogenic exposure while preserving nutritional quality. Local food plants that express protein fragments of pathogens might provide an effective means to stimulate gut mucosal immunity while increasing vaccine accessibility. Publication Types: Review PMID: 16566866 [PubMed - indexed for MEDLINE] 764: Microb Cell Fact. 2006 Mar 23;5:14. Heterologous expression of Brucella abortus GroEL heat-shock protein in Lactococcus lactis. Miyoshi A, Bermúdez-Humarán LG, Ribeiro LA, Le Loir Y, Oliveira SC, Langella P, Azevedo V. Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte - MG, Brasil. miyoshi@icb.ufmg.br BACKGROUND: Brucella abortus is a facultative intracellular pathogen that mainly infects cattle and humans. Current vaccines rely on live attenuated strains of B. abortus, which can revert to their pathogenic status and thus are not totally safe for use in humans. Therefore, the development of mucosal live vaccines using the food-grade lactic acid bacterium, Lactococcus lactis, as an antigen delivery vector, is an attractive alternative and a safer vaccination strategy against B. abortus. Here, we report the construction of L. lactis strains genetically modified to produce B. abortus GroEL heat-shock protein, a candidate antigen, in two cellular locations, intracellular or secreted. RESULTS: Only the secreted form of GroEL was stably produced in L. lactis, suggesting a detrimental effect of GroEL protein when intracellularly produced in this bacterium. Only trace amounts of mature GroEL were detected in the supernatant fraction of induced lactococcal cultures, and the GroEL precursor remained stacked in the cell fraction. Attempts to raise the secretion yields were made, but even when GroEL was fused to a synthetic propeptide, secretion of this antigen was not improved. CONCLUSION: We found that L. lactis is able to produce, and to secrete, a stable form of GroEL into the extracellular medium. Despite the low secretion efficiency of GroEL, which suggest that this antigen interacts with the cell envelope of L. lactis, secretion seems to be the best way to achieve both production and protein yields, regardless of cellular location. The L. lactis strain secreting GroEL has potential for in vivo immunization. PMID: 16556312 [PubMed] 765: Planta. 2006 May;223(6):1115-22. Epub 2006 Mar 23. Contribution of the arbuscular mycorrhizal symbiosis to heavy metal phytoremediation. Göhre V, Paszkowski U. Department of Molecular Biology and Plant Biology, University of Geneva, 30 Quai Ernest Ansermet, CH-1211, Geneva 4, Switzerland. High concentrations of heavy metals (HM) in the soil have detrimental effects on ecosystems and are a risk to human health as they can enter the food chain via agricultural products or contaminated drinking water. Phytoremediation, a sustainable and inexpensive technology based on the removal of pollutants from the environment by plants, is becoming an increasingly important objective in plant research. However, as phytoremediation is a slow process, improvement of efficiency and thus increased stabilization or removal of HMs from soils is an important goal. Arbuscular mycorrhizal (AM) fungi provide an attractive system to advance plant-based environmental clean-up. During symbiotic interaction the hyphal network functionally extends the root system of their hosts. Thus, plants in symbiosis with AM fungi have the potential to take up HM from an enlarged soil volume. In this review, we summarize current knowledge about the contribution of the AM symbiosis to phytoremediation of heavy metals. Publication Types: Review PMID: 16555102 [PubMed - indexed for MEDLINE] 766: New Genet Soc. 2005;24(2):139-55. In the democracies of DNA: ontological uncertainty and political order in three states. Jasanoff S. Harvard University, John F. Kennedy School of Government, 79 JFK Street, Cambridge, MA 02138, USA. sheila_jasanoff@harvard.edu This paper compares the regulation of biotechnology in Britain, Germany and the United States and shows that systematic differences have developed around four issues: abortion, assisted reproduction, stem cells, and genetically modified crops and foods. Policy choices with respect to these issues reflect the capacity of each nation's regulatory institutions to deal with the scientific, social and ethical uncertainties around biotechnology. National regulatory frameworks constitute an apparatus of collective sense-making through which governments and publics interpret biotechnology's risks and promises. Specifically, regulatory choices position the novel ontologies created by biotechnology either on the side of the familiar and manageable or on the side of the unknown and insupportably risky. The comparison shows that public responses to biotechnology are embedded within robust and coherent political cultures and are not ad hoc expressions of concern that very unpredictably from issue to issue. Publication Types: Comparative Study PMID: 16552932 [PubMed - indexed for MEDLINE] 767: New Genet Soc. 2005 Apr;24(1):31-56. Attitudes to biotechnology: estimating the opinions of a better-informed public. Sturgis P, Cooper H, Fife-Schaw C. Department of Sociology, School of Human Sciences, University of Surrey, Guildford, GU2 7XH, UK. p.sturgis@surrey.ac.uk Public familiarity with basic scientific concepts and principles has been proposed as essential for effective democratic decision-making (Miller, 1998). Empirical research, however, finds that public 'scientific literacy' is generally low, falling well short of what normative criteria would consider 'acceptable.' This has prompted calls to better engage, educate and inform the public on scientific matters, with the additional, usually implicit assumption that a knowledgeable citizenry should express more supportive and favourable attitudes toward science. Research investigating the notion that 'to know science is to love it' has provided only weak empirical support and has itself been criticised for representing science and technology as a unified and homogenous entity. In practice, it is argued, how knowledge impacts on the favourability of attitudes will depend on a multiplicity of actors, not the least of which is the particular area of science in question and the technologies to which it gives rise (Evans & Durant, 1992). This article uses a new method for examining the knowledge-attitude nexus on a prominent area of 21st century science--biotechnology. The idea that greater scientific knowledge can engender change in the favourability of attitudes toward specific areas of science is investigated using data from the 2000 British Social Attitudes Survey and the 1999 Wellcome Consultative Panel on Gene Therapy. Together the surveys measure public opinion on particular applications of genetic technologies, including gene therapy and the use of genetic data, as well as more general attitudes towards genetic research. We focus our analysis on how two different measures of knowledge impact on these attitudes; one a more general measure of scientific knowledge, the other relating specifically to knowledge of modern genetic science. We investigate what impact these knowledge domains have on attitudes toward biotechnology using a regression-based modelling technique (Bartels, 1996; Althaus, 1998; Sturgis, 2003). Controlling for a range of socio-demographic characteristics, we provide estimates of what collective and individual opinion would look like if everyone were as knowledgeable as the currently best-informed members of the general public on the knowledge domains in question. Our findings demonstrate that scientific knowledge does appear to have an important role in determining individual and group attitudes to genetic science. However, we find no support for a simple 'deficit model' of public understanding, as the nature of the relationship itself depends on the application of biotechnology in question and the social location of the individual. Publication Types: Research Support, Non-U.S. Gov't PMID: 16552916 [PubMed - indexed for MEDLINE] 768: J Agric Environ Ethics. 2005;18(5):495-508. Measuring biotechnology employees' ethical attitudes towards a controversial transgenic cattle project: the ethical valance matrix. Small BH, Fisher MW. Social Research Unit, AgResearch Ltd, Ruakura Research Centre, Hamilton, New Zealand. bruce.small@agresearch.co.nz What is the relationship between biotechnology employees' beliefs about the moral outcomes of a controversial transgenic research project and their attitudes of acceptance towards the project? To answer this question, employees (n=466) of a New Zealand company, AgResearch Ltd., were surveyed regarding a project to create transgenic cattle containing a synthetic copy of the human myelin basic protein gene (hMBP). Although diversity existed amongst employees' attitudes of acceptance, they were generally: in favor of the project, believed that it should be allowed to proceed to completion, and that it is acceptable to use transgenic cattle to produce medicines for humans. These three items were aggregated to form a project acceptance score. Scales were developed to measure respondents' beliefs about the moral outcomes of the project for identified stakeholders in terms of the four principles of common morality (benefit, non-harm, justice, and autonomy). These data were statistically aggregated into an Ethical Valence Matrix fo the project. The respondents' project Ethical Valence Scores correlated significantly with their project acceptance scores (r=0.64, p<0.001), accounting for 41% of the variance in respondents' acceptance attitudes. Of the four principles, non-harm had the strongest correlation with attitude to the project (r=0.59), followed by benefit and justice (both r=0.54), then autonomy (r=0.44). These results indicate that beliefs about the moral outcomes of a research project, in terms of the four principles approach, are strongly related to, and may be significant determinants of, attitudes to the research project. This suggests that, for employees of a biotechnology organization, ethical reasoning could be a central mechanism for the evaluation of the acceptability of a project. We propose that the Ethical Valence Matrix may be used as a tool to measure ethical attitudes towards controversial issues, providing a metric for comparison of perceived ethical consequences for multiple stakeholder groups and for the evaluation and comparison of the ethical consequences of competing alternative issues or projects. The tool could be used to measure both public and special interest groups' ethical attitudes and results used for the development of socially responsible policy or by science organizations as a democratizing decision aid to selection amongst projects competing for scarce research funds. PMID: 16552906 [PubMed - indexed for MEDLINE] 769: Appetite. 2006 May;46(3):324-31. Epub 2006 Mar 20. Attitudes towards genetically modified and organic foods. Saher M, Lindeman M, Hursti UK. Department of Psychology, University of Helsinki, P.O. Box 9, 00014 Helsinki, Finland. marieke.saher@helsinke.fi Finnish students (N=3261) filled out a questionnaire on attitudes towards genetically modified and organic food, plus the rational-experiential inventory, the magical thinking about food and health scale, Schwartz's value survey and the behavioural inhibition scale. In addition, they reported their eating of meat. Structural equation modelling of these measures had greater explanatory power for attitudes towards genetically modified (GM) foods than for attitudes towards organic foods (OF). GM attitudes were best predicted by natural science education and magical food and health beliefs, which mediated the influence of thinking styles. Positive attitudes towards organic food, on the other hand, were more directly related to such individual differences as thinking styles and set of values. The results of the study indicate that OF attitudes are rooted in more fundamental personal attributes than GM attitudes, which are embedded in a more complex but also in a more modifiable network of characteristics. Publication Types: Research Support, Non-U.S. Gov't PMID: 16546293 [PubMed - indexed for MEDLINE] 770: Vaccine. 2006 May 1;24(18):3900-8. Epub 2006 Mar 2. Immunogenicity of a malaria parasite antigen displayed by Lactococcus lactis in oral immunisations. Ramasamy R, Yasawardena S, Zomer A, Venema G, Kok J, Leenhouts K. BioMaDe Technology, Nijenborgh 4, 9747 AG Groningen, The Netherlands. A putative protective protein from Plasmodium falciparum merozoites, MSA2, was expressed in two different ways on the cell surface of the Gram-positive food-grade bacterium, Lactococcus lactis. The first display format exploits an LPXTG-type anchoring motif of the lactococcal proteinase PrtP to covalently anchor MSA2 to the genetically modified producer cells. In a second display format, MSA2 was fused to the peptidoglycan-binding domain (Protein Anchor) of the lactococcal cell wall hydrolase AcmA and was non-covalently rebound to the surface of non-genetically modified, non-living high-binder L. lactis cells, termed Gram-positive enhancer matrix (GEM) particles. The L. lactis recombinants carrying covalently bound MSA2 were used to immunise rabbits through nasal and oral routes. The highest levels of IgG antibodies reacting with near-native MSA2 on merozoites was elicited by oral administration. Intestinal antibodies to MSA2 were produced only after oral immunisation. MSA2-specific T(h)-cell activation could be demonstrated. Based on these results, the immunogenicity in oral immunisations of MSA2, bound non-covalently to non-genetically modified L. lactis GEM particles, was compared with MSA2 that was bound covalently to genetically modified L. lactis. These two forms elicited similar titres of serum antibodies. The results illustrate the potential of using non-genetically modified L. lactis as a safe vaccine delivery vehicle to elicit systemic antibodies, thereby avoiding the dissemination of recombinant DNA into the environment. PMID: 16545511 [PubMed - indexed for MEDLINE] 771: Kokuritsu Iyakuhin Shokuhin Eisei Kenkyusho Hokoku. 2005;(123):1-11. [Establishment of standards and specifications for chemical substances in foods and evaluation of exposure to maintain food safety] [Article in Japanese] Maitani T. Division of Foods, National Institute of Health Sciences. maitani@nihs.go.jp Currently, consumers are very anxious about many chemical substances contained in foods. To maintain food safety, the Ministry of Health, Labour and Welfare of Japan establishes standards and specifications on toxic chemical substances in foods, establishes analytical methods for surveillance, and investigates the daily dietary intake of food contaminants every year. This paper describes what sorts of standards and specifications for toxic chemical substances in foods have been established and what kinds of research on daily dietary intake have been performed. As the subjects for description, pesticide residues, toxic metals, dioxins, acrylamide, food additives, genetically modified food products, so-called health foods, and food allergens are included. Publication Types: English Abstract PMID: 16541744 [PubMed - indexed for MEDLINE] 772: Curr Opin Biotechnol. 2006 Apr;17(2):179-82. Epub 2006 Mar 15. Molecular advances and novel directions in food biotechnology innovation. Kleerebezem M. Publication Types: Editorial Review PMID: 16540309 [PubMed - indexed for MEDLINE] 773: J Dairy Sci. 2006 Apr;89(4):1254-66. Major advances associated with reproduction in dairy cattle. Moore K, Thatcher WW. Department of Animal Sciences, University of Florida, Gainesville, 32611-0910, USA. The purpose of this overview is to review some of the major advances in reproductive technologies, and how they may be applied to meet the challenge of enhancing reproductive efficiency in the high-producing dairy cow of the 21st century. The current population of high-producing dairy cows is considered to be subfertile, as characterized by low pregnancy rates and high rates of embryonic mortality. Coordinated systems of reproductive management have been developed based upon a thorough understanding of the endocrine, cellular, and molecular factors controlling ovarian and uterine function. These systems will partially restore herd reproductive performance. Advances in other reproductive technologies offer possibilities for wider use of superior germplasm. Technologies such as sexed semen, cloning, transgenesis, and preimplantation genetic diagnosis offer the potential to enhance the influence of superior animals on production of food for human consumption. However, at this time, additional research is needed to counteract the higher rates of embryonic and fetal mortality associated with some of these technologies. Furthermore, use of genomics, proteomics, and bioinformatics in the study of reproduction will undoubtedly provide investigators with a greater understanding of the limitations to efficient reproductive processes in the subfertile lactating dairy cow. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 16537958 [PubMed - indexed for MEDLINE] 774: Proc Natl Acad Sci U S A. 2006 Mar 7;103(10):3546-51. Epub 2006 Feb 28. High-amylose wheat generated by RNA interference improves indices of large-bowel health in rats. Regina A, Bird A, Topping D, Bowden S, Freeman J, Barsby T, Kosar-Hashemi B, Li Z, Rahman S, Morell M. Commonwealth Scientific and Industrial Research Organization, Food Futures National Research Flagship, P.O. Box 93, North Ryde 1670, NSW, Australia. Foods high in resistant starch have the potential to improve human health and lower the risk of serious noninfectious diseases. RNA interference was used to down-regulate the two different isoforms of starch-branching enzyme (SBE) II (SBEIIa and SBEIIb) in wheat endosperm to raise its amylose content. Suppression of SBEIIb expression alone had no effect on amylose content; however, suppression of both SBEIIa and SBEIIb expression resulted in starch containing >70% amylose. When the >70% amylose wheat grain was fed to rats in a diet as a wholemeal, several indices of large-bowel function, including short-chain fatty acids, were improved relative to standard wholemeal wheat. These results indicate that this high-amylose wheat has a significant potential to improve human health through its resistant starch content. Publication Types: Research Support, Non-U.S. Gov't PMID: 16537443 [PubMed - indexed for MEDLINE] 775: J Agric Food Chem. 2006 Mar 22;54(6):2169-77. Application of two-dimensional gel electrophoresis to interrogate alterations in the proteome of gentically modified crops. 3. Assessing unintended effects. Ruebelt MC, Lipp M, Reynolds TL, Schmuke JJ, Astwood JD, DellaPenna D, Engel KH, Jany KD. Product Safety Center, Monsanto Company, 800 North Lindbergh Boulevard, St. Louis, Missouri 63167, USA. m.ruebelt@gmail.com The current procedures to assess the safety of food and feed derived from modern biotechnology include the investigation of possible unintended effects. To improve the probability of detecting unintended effects, profiling techniques such as proteomics are currently tested as complementary analytical tools to the existing safety assessment. An optimized two-dimensional gel electrophoresis (2DE) method was used as a proteomics approach to investigate insertional and pleiotropic effects on the proteome due to genetic engineering. Twelve transgenic Arabidopsis thaliana lines were analyzed by 2DE, and their seed proteomes were compared to that of their parental line as well as to 12 Arabidopsis ecotype lines. The genetic modification of the Arabidopsis lines, using three different genes and three different promoters, did not cause unintended changes to the analyzed seed proteome. Differences in spot quantity between transgenic and nontransgenic lines fell in the range of values found in the 12 Arabidopsis ecotype lines or were related to the introduced gene. Publication Types: Comparative Study PMID: 16536592 [PubMed - indexed for MEDLINE] 776: J Agric Food Chem. 2006 Mar 22;54(6):2154-61. Application of two-dimensional gel electrophoresis to interrogate alterations in the proteome of genetically modified crops. 1. Assessing analytical validation. Ruebelt MC, Leimgruber NK, Lipp M, Reynolds TL, Nemeth MA, Astwood JD, Engel KH, Jany KD. Product Safety Center, Monsanto Company, 800 North Lindbergh Boulevard, St. Louis, Missouri 63167, USA. m.ruebelt@gmail.com Current tools used to assess the safety of food and feed derived from modern biotechnology emphasize the investigation of possible unintended effects caused directly by the expression of transgenes or indirectly by pleiotropy. These tools include extensive multisite and multiyear agronomic evaluations, compositional analyses, animal nutrition, and classical toxicology evaluations. Because analytical technologies are rapidly developing, proteome analysis based on two-dimensional gel electrophoresis (2DE) was investigated as a complementary tool to the existing technologies. A 2DE method was established for the qualitative and quantitative analysis of the seed proteome of Arabidopsis thaliana with the following validation parameters examined: (1) source and scope of variation; (2) repeatability; (3) sensitivity; and (4) linearity of the method. The 2DE method resolves proteins with isoelectric points between 4 and 9 and molecular masses (MM) of 6-120 kDa and is sensitive enough to detect protein levels in the low nanogram range. The separation of the proteins was demonstrated to be very reliable with relative position variations of 1.7 and 1.1% for the pI and MM directions, respectively. The mean coefficient of variation of 254 matched spot qualities was found to be 24.8% for the gel-to-gel and 26% for the overall variability. A linear relationship (R2 > 0.9) between protein amount and spot volume was demonstrated over a 100-fold range for the majority of selected proteins. Therefore, this method could be used to interrogate proteome alterations such as a novel protein, fusion protein, or any other change that affects molecular mass, isoelectric point, and/or quantity of a protein. PMID: 16536590 [PubMed - indexed for MEDLINE] 777: J Agric Food Chem. 2006 Mar 22;54(6):2145-53. Molecular characterization of a stable antisense chalcone synthase phenotype in strawberry (Fragaria x ananassa). Lunkenbein S, Coiner H, de Vos CH, Schaart JG, Boone MJ, Krens FA, Schwab W, Salentijn EM. Biomolecular Food Technology, Technical University München, Lise-Meitner-Strasse 34, 85354 Freising, Germany. An octaploid (Fragaria x ananassa cv. Calypso) genotype of strawberry was transformed with an antisense chalcone synthase (CHS) gene construct using a ripening related CHS cDNA from Fragaria x ananassa cv. Elsanta under the control of the constitutive CaMV 35S promoter via Agrobacterium tumefaciens. Out of 25 transgenic lines, nine lines showed a reduction in CHS mRNA accumulation of more than 50% as compared to the untransformed cv. Calypso control. The antisense CHS construct was found to be integrated into the genome, with a copy number ranging from one to four. The pigmentation of the fruit was only affected when less than 5% of the control CHS expression level was detected. A stable antisense phenotype over a period of 4 years was obtained in the primary transgenic lines at a rate of 1:20. As a consequence of the reduced activity of CHS, the levels of anthocyanins, flavonols, and proanthocyanidins were downregulated and precursors of the flavonoid pathway were shunted to the phenylpropanoid pathway leading to highly increased levels of cinnamoyl glucose (520% of control), caffeoyl glucose (816% of control), and feruloyl glucose (1092% of control) as well as p-coumaryl alcohol (363% of control) and p-coumaryl-1-acetate (1079% of control), which occur only as trace components in untransformed control fruits. These results demonstrate that the introduction of an antisense CHS construct in strawberry results in an unpredictable biochemical phenotype, thereby confirming that CHS function is an important regulatory point of substrate flow between the flavonoid and the phenylpropanoid pathways. PMID: 16536589 [PubMed - indexed for MEDLINE] 778: Wei Sheng Yan Jiu. 2005 Nov;34(6):732-4. [Detection of genetically modified organisms in food and animal feed by polymerase chain reaction] [Article in Chinese] Zhou JC, Yang MJ, Yang XF, Huang JM. Toxicological Laboratory, Guangdong Center for Disease Control and Prevention, Guangzhou 510300, China. OBJECTIVE: To investigate the presence of genetically modified organisms (GMO) in the foods and animal feed samples in Guangzhou market. METHODS: The presence of GMO were investigated by PCR detection of camv 35S promoter and nos terminator, and the presence of RoundUp Ready Soybean (RRS), Bt176 Maximaizer or Mon810 YieldGard in GMO-positive samples were further determined by PCR detecting their specific DNA fragments respectively. RESULTS: One corn soup sample, two soybean samples, one potato fries sample as well as two animal feed samples were revealed to be GMO-positive in twenty-two food samples and three animal feed samples, and the presence of RRS in the GMO-positive soybean samples and the two positive animal feed samples were verified by PCR detection of a 129 bp RRS-specific DNA fragment, however, no Bt176 Maximaizer or Mon810 YieldGard specific PCR products were obtained with the GMO-positive corn soup and animal feed DNA samples used as PCR templates. CONCLUSION: Genetically modified organism presented in foods and animal feeds even though they were not been labelled. Publication Types: English Abstract Research Support, Non-U.S. Gov't PMID: 16535848 [PubMed - in process] 779: J Zhejiang Univ Sci B. 2006 Apr;7(4):257-66. Chinese public understanding of the use of agricultural biotechnology--a case study from Zhejiang Province of China. Lü L. Department of Social Sciences, School of Humanities, Hangzhou Dianzi University, Hangzhou 310018, China. lulandk27@yahoo.com This study explores the Chinese public's perceptions of, and attitudes to, agriculture and food applications of biotechnology; and investigates the effect of socio-demographic factors on attitudes. A questionnaire survey and interviews were used in an attempt to combine quantitative analysis with qualitative review. The main finding of this study is that the Chinese population has a superficial, optimistic attitude to agricultural biotechnology; and that, in accordance with public attitudes, a cautious policy, with obligatory labelling, should be adopted. The study reveals that education is the factor among socio-demographic variables with the strongest impact on public attitudes. Higher education leads to a more positive evaluation of GM (genetically modified) foods and applications of biotechnology with respect to usefulness, moral acceptability, and suitability for encouragement. In addition, public attitudinal differences depend significantly on area of residence. Compared with their more urban compatriots, members of the public in less developed areas of China have more optimistic attitudes, perceive more benefits, and are more risk tolerant in relation to GM foods and agricultural biotechnology. Finally we obtained a very high rate of "don't know" answers to our survey questions. This suggests that many people do not have settled attitudes, and correspondingly, that the overall public attitude to agricultural biotechnology and GM foods in China is at present somewhat unstable. Publication Types: Research Support, Non-U.S. Gov't PMID: 16532526 [PubMed - indexed for MEDLINE] 780: Environ Geochem Health. 2006 Feb-Apr;28(1-2):103-10. Transgenic Spartina alterniflora for phytoremediation. Czakó M, Feng X, He Y, Liang D, Márton L. Department of Biological Sciences, University of South Carolina, 700 Sumter Street, Columbia, SC 29208, USA. czako@mail.biol.sc.edu Perennial monoculture forming grasses are very important natural remediators of pollutants. Their genetic improvement is an important task because introduction of key transgenes can dramatically improve their remediation potential. Transfer of key genes for mercury phytoremediation into the salt marsh cordgrass (Spartina alterniflora) is reported here. S. alterniflora plays an important role in the salt marsh by cycling of elements, both nutrients and pollutants, protects the coastline from erosion, is a keystone species in the salt marsh supporting a large food web, which in turn supports a significant segment of economy, including tourism, has an impact on cloud formation and consequently on global weather, and is thus an ecologically important species relevant for our life-support systems. Embryogenic callus of S. alterniflora was co-inoculated with a pair of Agrobacterium strains LBA4404 carrying the organomercurial lyase (merB) and mercuric reductase (merA) genes, respectively, in order to co-introduce both the merA and the merB genes. Seven stable geneticin resistant lines were recovered. The presence of merA and merB genes was verified by PCR and Southern blotting. All but one transgenic lines contained both the merA and the merB sequences proving that co-introduction into Spartina of two genes from separate Agrobacterium strains is feasible and frequent, although the overall frequency of transformation is low. Northern blotting showed differences in relative expression of the two transgenes among individual transformants. The steady-state RNA levels appeared to correlate with the phenotype. Line #7 showed the highest resistance to HgCl(2) (up to 500 microM), whereas line #3 was the most resistant to phenylmercuric acetate (PMA). Wild-type (WT) callus is sensitive to PMA at 50 microM and to HgCl(2) at 225 microM. Publication Types: Research Support, Non-U.S. Gov't PMID: 16528587 [PubMed - indexed for MEDLINE] 781: Plant Mol Biol. 2006 Mar;60(4):555-63. Generation of phenylpropanoid pathway-derived volatiles in transgenic plants: rose alcohol acetyltransferase produces phenylethyl acetate and benzyl acetate in petunia flowers. Guterman I, Masci T, Chen X, Negre F, Pichersky E, Dudareva N, Weiss D, Vainstein A. The Robert H. Smith Institute of Plant Sciences and Genetics in Agriculture, Faculty of Agricultural, Food and Environmental Quality Sciences, The Hebrew University of Jerusalem, Rehovot 76100, Israel. Esters are important contributors to the aroma of numerous flowers and fruits. Acetate esters such as geranyl acetate, phenylethyl acetate and benzyl acetate are generated as a result of the action of alcohol acetyltransferases (AATs). Numerous homologous AATs from various plants have been characterized using in-vitro assays. To study the function of rose alcohol acetyltransferase (RhAAT) in planta, we generated transgenic petunia plants expressing the rose gene under the control of a CaMV-35S promoter. Although the preferred substrate of RhAAT in vitro is geraniol, in transgenic petunia flowers, it used phenylethyl alcohol and benzyl alcohol to produce the corresponding acetate esters, not generated by control flowers. The level of benzyl alcohol emitted by the flowers of different transgenic lines was ca. three times higher than that of phenylethyl alcohol, which corresponded to the ratio between the respective products, i.e. ca. three times more benzyl acetate than phenylethyl acetate. Feeding of transgenic petunia tissues with geraniol or octanol led to the production of their respective acetates, suggesting the dependence of volatile production on substrate availability. Publication Types: Research Support, Non-U.S. Gov't PMID: 16525891 [PubMed - indexed for MEDLINE] 782: J Chem Ecol. 2006 Jan;32(1):1-13. Epub 2006 Feb 26. Potato expressing beetle-specific Bacillus thuringiensis Cry3Aa toxin reduces performance of a moth. Hussein HM, Habustová O, Turanli F, Sehnal F. Institute of Entomology, Academy of Sciences, Branisovská 31, 370 05, Ceské Budejovice, Czech Republic. Expression of the Bacillus thuringiensis beetle-specific toxin Cry3Aa, which renders a genetically modified potato cultivar resistant to the Colorado potato beetle Leptinotarsa decemlineata, exerts a deleterious effect on the polyphagous moth Spodoptera littoralis. The caterpillars of S. littoralis feed less and produce smaller pupae on the genetically modified cultivar (NewLeaf Superior) than on the parental nontransgenic cultivar (Superior). The conversion efficiencies of total dry matter, combustion heat, carbon, and nitrogen from leaves to insect biomass are similar on both cultivars. In spite of similar food utilization and a relatively small difference in the body mass at pupation, female adults that developed from caterpillars fed on NewLeaf Superior lay a mean of 309 eggs compared to a mean of 713 eggs deposited by females that developed from caterpillars fed on Superior. Because of this difference and a simultaneous reduction in fertility (egg hatchability) from 78 to 48%, a pair of adults that fed as larvae on NewLeaf Superior produces only 148 larvae, whereas a pair of adults that fed as larvae on Superior produces 556 larvae. We suggest that small amounts of Cry3Aa that accumulate in insect tissue and persist until the adult stage are responsible for the decline in reproduction. Publication Types: Research Support, Non-U.S. Gov't PMID: 16525866 [PubMed - indexed for MEDLINE] 783: J Sep Sci. 2006 Feb;29(2):197-210. High-performance liquid chromatography and capillary electrophoresis for the analysis of maize proteins. Rodriguez-Nogales JM, Garcia MC, Marina ML. Departamento de Química Analítica, Universidad de Alcalá, Madrid, Spain. Methods for the analysis of maize proteins using HPLC and CE are reviewed. Most of the references cited in this review concern HPLC methods. Size-exclusion HPLC and especially RP-HPLC methods have been developed for characterization of normal and genetically modified maize, cultivar differentiation, and prediction of quality. Few CE methods for the analysis of maize proteins were found in the existing literature. Most of these methods focus on optimization of the separation of maize proteins using CZE and SDS-capillary gel electrophoresis. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 16524093 [PubMed - indexed for MEDLINE] 784: McGeorge Law Rev. 2000;32(1):89-110. Biotechnology and the creation of ethics. Coletta RR. University of the Pacific, McGeorge School of Law, USA. PMID: 16523579 [PubMed - indexed for MEDLINE] 785: Curr Opin Biotechnol. 2006 Apr;17(2):174-8. Epub 2006 Mar 7. Economic impact of transgenic crops in developing countries. Raney T. Food and Agriculture Organization of the United Nations, Viale delle Terme di Caracalla, 00100 Rome, Italy. terri.raney@fao.org Transgenic crops are being adopted rapidly at the global level, but only a few developing countries are growing them in significant quantities. Why are these crops so successful in some countries but not in others? Farm level profitability ultimately determines whether farmers adopt and retain a new technology, but this depends on much more than technical performance. Recent economic studies in developing countries find positive, but highly variable, economic returns to adopting transgenic crops. These studies confirm that institutional factors such as national agricultural research capacity, environmental and food safety regulations, intellectual property rights and agricultural input markets matter at least as much as the technology itself in determining the level and distribution of economic benefits. Publication Types: Review PMID: 16522366 [PubMed - indexed for MEDLINE] 786: Allergy. 2006 Apr;61(4):491-7. Allergenicity assessment of transgenic mustard (Brassica juncea) expressing bacterial codA gene. Singh AK, Mehta AK, Sridhara S, Gaur SN, Singh BP, Sarma PU, Arora N. Institute of Genomics and Integrative Biology, Delhi, India. BACKGROUND: Assessing the allergenicity and toxicity of genetically modified (GM) crops is essential before they become a regular part of our food supply. The present study aimed to assess the allergenicity of Brassica juncea (mustard) expressing choline oxidase (codA) gene from Arthrobacter globiformis that provides resistance against abiotic stresses. METHODS: SDAP, Farrp, and Swiss-Prot databases were used to study allergenicity of choline oxidase. Digestibility of choline oxidase was assessed in simulated gastric fluid (SGF). Specific immunoglobulin E (IgE) reactivity of native and GM mustard was compared by using enzyme-linked immunosorbent assay (ELISA) and skin tests in respiratory-allergic patients. Allergenicity of GM and native mustard proteins was compared in Balb/c mice. RESULTS: Choline oxidase showed no significant homology with allergenic proteins in SDAP and Farrp databases. Cross-reactive epitope search showed a stretch similar to Hev b 6 having some antigenic properties. Purified choline oxidase showed complete degradation with SGF. Skin prick test of native and GM mustard extract on respiratory allergic patients showed significant correlation (P < 0.05). ELISA with 96 patients' sera showed comparable IgE reactivity. Balb/c mice immunized with native and GM mustard proteins showed low IgE response. Presensitized mice on intravenous challenge with Brassica extract showed no anaphylactic symptoms unlike ovalbumin (OVA) sensitization that showed anaphylactic reaction in mice. Lung histology of OVA-sensitized mice showed narrowing of airway and large eosinophilic infiltration, whereas native and GM Brassica extract showed normal airway. CONCLUSION: Genetically modified mustard with the codA gene possessed allergenicity similar to that of native mustard and no enhancement of IgE binding was observed due to genetic manipulation. Publication Types: Research Support, Non-U.S. Gov't PMID: 16512812 [PubMed - indexed for MEDLINE] 787: J AOAC Int. 2006 Jan-Feb;89(1):232-9. Scoring in genetically modified organism proficiency tests based on log-transformed results. Thompson M, Ellison SL, Owen L, Mathieson K, Powell J, Key P, Wood R, Damant AP. University of London, Birkbeck College, School of Biological and Chemical Sciences, Malet St, London, United Kingdom. The study considers data from 2 UK-based proficiency schemes and includes data from a total of 29 rounds and 43 test materials over a period of 3 years. The results from the 2 schemes are similar and reinforce each other. The amplification process used in quantitative polymerase chain reaction determinations predicts a mixture of normal, binomial, and lognormal distributions dominated by the latter 2. As predicted, the study results consistently follow a positively skewed distribution. Log-transformation prior to calculating z-scores is effective in establishing near-symmetric distributions that are sufficiently close to normal to justify interpretation on the basis of the normal distribution. Publication Types: Research Support, Non-U.S. Gov't PMID: 16512253 [PubMed - indexed for MEDLINE] 788: Curr Opin Biotechnol. 2006 Apr;17(2):130-8. Epub 2006 Feb 28. Metabolic engineering in plants for human health and nutrition. Kinney AJ. Crop Genetics Research, DuPont Experimental Station, Wilmington, DE 19880-0353, USA. anthony.kinney@cgr.dupont.com In many cases, multiple pathway enzymes need to be upregulated to produce a significant yield of a desired product. Technical advances in simultaneously manipulating multiple steps in plant metabolic pathways include the use of transcription factors, such as MYB12. By upregulating the genes of an entire pathway, these factors can greatly simplify multienzyme engineering. Furthermore, synthetic zinc-finger protein transcription factors can now be designed to target specific pathway enzymes, such as tocopherol methyltransferases. When multiple steps in a pathway are upregulated, previously unsuspected facets of the pathway might be revealed, such as the newly uncovered bifunctional substrate preference of the key regulatory enzyme in tocopherol (vitamin E) biosynthesis, homogentisate phytyltransferase. The engineering of desired traits, such as long-chain omega-3 polyunsaturated fatty acids, can require entirely new pathways to be introduced into a plant. Recent advances in genomics and gene expression technology have made this type of complex metabolic engineering highly feasible. Publication Types: Review PMID: 16510274 [PubMed - indexed for MEDLINE] 789: Mol Cell Biol. 2006 Mar;26(6):2286-96. A family knockout of all four Drosophila metallothioneins reveals a central role in copper homeostasis and detoxification. Egli D, Yepiskoposyan H, Selvaraj A, Balamurugan K, Rajaram R, Simons A, Multhaup G, Mettler S, Vardanyan A, Georgiev O, Schaffner W. IMB Universität Zürich, Winterthurerstrasse 190, CH-8057 Zürich, Switzerland. Metallothioneins are ubiquitous, small, cysteine-rich proteins with the ability to bind heavy metals. In spite of their biochemical characterization, their in vivo function remains elusive. Here, we report the generation of a metallothionein gene family knockout in Drosophila melanogaster by targeted disruption of all four genes (MtnA to -D). These flies are viable if raised in standard laboratory food. During development, however, they are highly sensitive to copper, cadmium, and (to a lesser extent) zinc load. Metallothionein expression is particularly important for male viability; while copper load during development affects males and females equally, adult males lacking metallothioneins display a severely reduced life span, possibly due to copper-mediated oxidative stress. Using various reporter gene constructs, we find that different metallothioneins are expressed with virtually the same tissue specificity in larvae, notably in the intestinal tract at sites of metal accumulation, including the midgut's "copper cells." The same expression pattern is observed with a synthetic minipromoter consisting only of four tandem metal response elements. From these and other experiments, we conclude that tissue specificity of metallothionein expression is a consequence, rather than a cause, of metal distribution in the organism. The bright orange luminescence of copper accumulated in copper cells of the midgut is severely reduced in the metallothionein gene family knockout, as well as in mutants of metal-responsive transcription factor 1 (MTF-1), the main regulator of metallothionein expression. This indicates that an in vivo metallothionein-copper complex forms the basis of this luminescence. Strikingly, metallothionein mutants show an increased, MTF-1-dependent induction of metallothionein promoters in response to copper, cadmium, silver, zinc, and mercury. We conclude that free metal, but not metallothionein-bound metal, triggers the activation of MTF-1 and that metallothioneins regulate their own expression by a negative feedback loop. Publication Types: Research Support, Non-U.S. Gov't PMID: 16508004 [PubMed - indexed for MEDLINE] 790: Environ Health Perspect. 2006 Mar;114(3):A146-7. Comment on: Environ Health Perspect. 2005 Aug;113(8):A526-33. The hazards of genetically engineered foods. Margulis C. Publication Types: Comment Letter PMID: 16507441 [PubMed - indexed for MEDLINE] 791: J Agric Food Chem. 2006 Mar 8;54(5):1699-709. Detection of transgenic and endogenous plant DNA in digesta and tissues of sheep and pigs fed Roundup Ready canola meal. Sharma R, Damgaard D, Alexander TW, Dugan ME, Aalhus JL, Stanford K, McAllister TA. Agriculture and Agri-Food Canada Research Centres, Lethbridge, Alberta, Canada. The persistence of plant-derived recombinant DNA in sheep and pigs fed genetically modified (Roundup Ready) canola was assessed by PCR and Southern hybridization analysis of DNA extracted from digesta, gastrointestinal (GI) tract tissues, and visceral organs. Sheep (n = 11) and pigs (n = 36) were fed to slaughter on diets containing 6.5 or 15% Roundup Ready canola. Native plant DNA (high- and low-copy-number gene fragments) and the cp4 epsps transgene that encodes 5-enolpyruvyl shikimate-3-phosphate synthase were tracked in ruminal, abomasal, and large intestinal digesta and in tissue from the esophagus, rumen, abomasum, small and large intestine, liver, and kidney of sheep and in cecal content and tissue from the duodenum, cecum, liver, spleen, and kidney of pigs. High-copy chloroplast-specific DNA (a 520-bp fragment) was detected in all digesta samples, the majority (89-100%) of intestinal tissues, and at least one of each visceral organ sample (frequencies of 3-27%) from sheep and swine. Low-copy rubisco fragments (186- and 540-bp sequences from the small subunit) were present at slightly lower, variable frequencies in digesta (18-82%) and intestinal tissues (9-27% of ovine and 17-25% of porcine samples) and infrequently in visceral organs (1 of 88 ovine samples; 3 of 216 porcine samples). Each of the five cp4 epsps transgene fragments (179-527 bp) surveyed was present in at least 27% of ovine large intestinal content samples (maximum = 64%) and at least 33% of porcine cecal content samples (maximum = 75%). In sheep, transgene fragments were more common in intestinal digesta than in ruminal or abomasal content. Transgene fragments were detected in 0 (esophagus) to 3 (large intestine) GI tract tissues from the 11 sheep and in 0-10 of the duodenal and cecal tissues collected from 36 pigs. The feed-ingested recombinant DNA was not detected in visceral tissues (liver, kidney) of lambs or in the spleen from pigs. Of note, however, one liver and one kidney sample from the pigs (different animals) were positive for a 278-bp fragment of the transgenic cp4 epsps (denoted F3). Examination of genomic libraries from these tissues yielded no conclusive information regarding integration of the fragment into porcine DNA. This study confirms that feed-ingested DNA fragments (endogenous and transgenic) do survive to the terminal GI tract and that uptake into gut epithelial tissues does occur. A very low frequency of transmittance to visceral tissue was confirmed in pigs, but not in sheep. It is recognized that the low copy number of transgenes in GM feeds is a challenge to their detection in tissues, but there was no evidence to suggest that recombinant DNA would be processed in the gut in any manner different from endogenous feed-ingested genetic material. Publication Types: Research Support, Non-U.S. Gov't PMID: 16506822 [PubMed - indexed for MEDLINE] 792: Food Chem Toxicol. 2006 Jul;44(7):1092-9. Epub 2006 Feb 17. Results of a 90-day safety assurance study with rats fed grain from corn borer-protected corn. Hammond BG, Dudek R, Lemen JK, Nemeth MA. Monsanto Company, Product Safety Center, 800 North Lindbergh Blvd., Bldg. O3F, St. Louis, MO 63167, USA. bruce.g.hammond@monsanto.com The results of a 90-day rat feeding study with grain from MON 810 corn (YieldGard Cornborer -- YieldGard Cornborer is a registered trademark of Monsanto Technology, LLC) that is protected against feeding damage from corn and stalk boring lepidopteran insects are presented. Corn borer protection was accomplished through the introduction of cry1Ab coding sequences into the corn genome for in planta production of a bioactive form of Cry1Ab protein. Grain from MON 810 and its near-isogenic control was separately formulated into rodent diets at levels of 11% and 33% (w/w) by Purina Mills, Inc. (PMI). All diets were nutritionally balanced and conformed to PMI specifications for Certified LabDiet (PMI Certified LabDiet 5002 is a registered trademark of Purina Mills, Inc.) 5002. There were a total of 400 rats in the study divided into 10 groups of 20 rats/sex/group. The responses of rats fed diets containing MON 810 were compared to those of rats fed grain from conventional corn varieties. Overall health, body weight, food consumption, clinical pathology parameters (hematology, blood chemistry, urinalysis), organ weights, and gross and microscopic appearance of tissues were comparable between groups fed diets containing MON 810 and conventional corn varieties. This study complements extensive agronomic, compositional and farm animal feeding studies with MON 810 grain, confirming that it is as safe and nutritious as grain from existing commercial corn varieties. PMID: 16487643 [PubMed - indexed for MEDLINE] 793: Curr Opin Biotechnol. 2006 Apr;17(2):198-203. Epub 2006 Feb 14. Food products and allergy development, prevention and treatment. Zuercher AW, Fritsché R, Corthésy B, Mercenier A. Allergy Group, Nutrition and Health Department, Nestlé Research Center, Vers-chez-les-Blanc, CH-1000 Lausanne 26, Switzerland. In westernized countries allergic diseases have reached epidemic proportions. Food is frequently a perpetrator of allergy but, in turn, modified food and selected food ingredients can become valuable intervention tools in the fight against allergy. There are two basic approaches towards mitigation of food allergy through nutrition: to reduce the allergenicity of raw food materials by physical, chemical or genetic methods or to influence host immunity towards a non-allergic state using various food ingredients. Dietary intervention for the prevention and therapy of allergy is an emerging field where initial findings from animal studies are now being validated in human trials. Nevertheless, to consolidate the utility of such interventions, more pre-clinical and clinical studies remain necessary. Publication Types: Review PMID: 16481157 [PubMed - indexed for MEDLINE] 794: J Anim Sci. 2006 Mar;84(3):597-607. The digestive fate of Escherichia coli glutamate dehydrogenase deoxyribonucleic acid from transgenic corn in diets fed to weanling pigs. Beagle JM, Apgar GA, Jones KL, Griswold KE, Radcliffe JS, Qiu X, Lightfoot DA, Iqbal MJ. Department of Animal Science, Food and Nutrition, Southern Illinois University, Carbondale, IL 62901, USA. Corn containing genetically engineered plasmid DNA encoding an Escherichia coli glutamate dehydrogenase (gdhA) was fed to 19-d-old weanling swine to trace the digestive fate of the transgenic DNA. Eight pens of 8 pigs were fed a commercial (nongdhA) starter for 2 wk. One pig was randomly selected from each pen for 0-h control samples. The remaining 56 pigs were transitioned onto a corn-soybean meal diet and fed a diet containing 58% gdhA corn for approximately 1 wk; immediately thereafter, liver, 10th rib muscle, white blood cells, and plasma from the hepatic portal vein and ingesta from the stomach, distal ileum, and large intestine were collected. The DNA was extracted and the concentration determined via spectrophotometry. Polymerase chain reaction and gel electrophoresis were performed with primers designed to amplify 490 bp that included the plasmid's ligation site between the maize ubiquitin and the gdhA genes. The gdhA corn-derived DNA and diet served as positive assay controls, and conventional corn DNA and distilled water acted as negative assay controls. Detection limits were 0.99 fg of target DNA confounded with 500 ng of conventional corn DNA per each 20 &L reaction. Transgenic DNA was detected in 71.43% of the stomach and 1.79% of the ileal ingesta samples from treatment animals but was not detected in the large intestine, white blood cells, plasma, liver, or muscle samples. Transgenic DNA was not detected in any sample from 0-h control animals. Stomach and ileal ingesta samples were further analyzed using real-time PCR. With an estimated limit of detection of 1.049 ag/microL, 89.29% of the stomach ingesta samples were positive (average 1.56 fg target DNA). The proportion of transgenic DNA to total DNA differed between diet and stomach ingesta samples (P < 0.001). Despite the greater sensitivity of real-time PCR, target DNA was detected in only 1.79% of ileal ingesta. These data suggest that the gdhA transgene began degradation in the stomach and was nondetectable in the large intestine. Publication Types: Comparative Study PMID: 16478951 [PubMed - indexed for MEDLINE] 795: J Agric Food Chem. 2006 Feb 22;54(4):1158-65. Real-time polymerase chain reaction (PCR) quantitative detection of Brassica napus using a locked nucleic acid TaqMan probe. Schmidt AM, Rott ME. Sidney Laboratory, Canadian Food Inspection Agency, 8801 East Saanich Road, Sidney, British Columbia V8L 1H3, Canada. Several countries have introduced mandatory labeling requirements on foods derived from genetically modified organisms. Real-time quantitative Polymerase Chain Reaction (PCR) has quickly become the method of choice in support of these regulations and requires the development of separate PCR assays targeting the transgenic sequence as well as a specific endogenous gene sequence. To develop a Brassica napus-specific PCR assay, partial sequences of the acetyl-CoA carboxylase BnACCg8 gene from B. napus and the closely related Brassica rapa were determined and compared, and a region of unique nucleotide sequence was identified. Universal amplification primers were designed to either side of this region, and a locked nucleic acid TaqMan probe was designed to the B. napus-specific sequence. Evaluation of this primer/probe combination indicated a high level of specificity to B. napus: no amplification signal was observed with any other species tested, including five closely related Brassica species. The method was assayed with 14 different B. napus cultivars, and comparable amplification curves were consistently obtained for all. The assay was highly sensitive, with a limit of detection between 1 and 10 haploid copies. Practically, the method was demonstrated to be effective for the detection of processed food samples and for the quantification of Roundup Ready canola content in mixed samples. Publication Types: Comparative Study PMID: 16478231 [PubMed - indexed for MEDLINE] 796: Appetite. 2006 Mar;46(2):144-51. Epub 2006 Feb 15. Awareness, acceptance of and willingness to buy genetically modified foods in Urban China. Huang J, Qiu H, Bai J, Pray C. Institute of Geographical Sciences and Natural Resource Research (IGSNRR), Chinese Academy of Sciences (CAS), Center for Chinese Agricultural Policy, Jia 11, Datun Road, Beijing 100101, China. jkhuang.ccap@igsnrr.ac.cn There is concern about the extent to which consumers will accept genetically modified (GM) foods if they are commercialized in China. The evidence from the existing literature is mixed and sometimes confusing. The objective of this study is to conduct a large in-depth face-to-face in-house survey that examines the consumers' awareness, acceptance of and willingness to buy GM foods in China. To achieve this objective, a well-designed consumer survey was conducted in 11 cities of five provinces in Eastern China in 2002 and 2003. The results indicate that despite much less information on GM foods available publicly in China, more than two thirds of consumers in urban areas have heard of GM foods. But their knowledge on biotechnology was limited. Chinese consumers' acceptance of and willingness to buy GM foods was much higher than in other countries. Chinese consumers also demonstrated great variance in their acceptance of different GM foods. Information and prices of GM foods were two important factors affecting consumers' attitudes toward GM foods. Based on the findings of this study and given that our sample is in the more developed eastern Urban China, we conclude that the commercialization of GM foods is not likely to receive great resistance from the consumers in China. Publication Types: Research Support, Non-U.S. Gov't PMID: 16469414 [PubMed - indexed for MEDLINE] 797: Toxicology. 2006 Apr 3;221(1):128-33. Epub 2006 Feb 8. cDNA microarray screening in food safety. Roy S, Sen CK. Laboratory of Molecular Medicine and DNA Microarray & Genetics Facility, Dorothy M. Davis Heart and Lung Research Institute, Department of Surgery, The Ohio State University Medical Center, Columbus, OH 43210, USA. The cDNA microarray technology and related bioinformatics tools presents a wide range of novel application opportunities. The technology may be productively applied to address food safety. In this mini-review article, we present an update highlighting the late breaking discoveries that demonstrate the vitality of cDNA microarray technology as a tool to analyze food safety with reference to microbial pathogens and genetically modified foods. In order to bring the microarray technology to mainstream food safety, it is important to develop robust user-friendly tools that may be applied in a field setting. In addition, there needs to be a standardized process for regulatory agencies to interpret and act upon microarray-based data. The cDNA microarray approach is an emergent technology in diagnostics. Its values lie in being able to provide complimentary molecular insight when employed in addition to traditional tests for food safety, as part of a more comprehensive battery of tests. Publication Types: Research Support, N.I.H., Extramural Review PMID: 16466843 [PubMed - indexed for MEDLINE] 798: Food Nutr Bull. 2005 Dec;26(4):443-4. Food biotechnology and consumer perceptions in Asia. Cairns G. Asian Food Information Network, Bangkok, Thailand. gcairns@afic.org Publication Types: Review PMID: 16465994 [PubMed - indexed for MEDLINE] 799: Food Nutr Bull. 2005 Dec;26(4):436-42. Nutritional and safety assessments of foods and feeds nutritionally improved through biotechnology. Chassy B, Hlywka JJ, Kleter GA, Kok EJ, Kuiper HA, McGloughlin M, Munro IC, Phipps RH, Reid JE, Stein J, Zabik J; Task Force for the International Life Sciences Institutue International Food Biotechnology Committee. University of Illinois, Urbana, USA. Publication Types: Review PMID: 16465993 [PubMed - indexed for MEDLINE] 800: Food Nutr Bull. 2005 Dec;26(4):432-5. Sharing Malaysian experience with the development of biotechnology-derived food crops. Abu Bakar UK, Pillai V, Hashim M, Daud HM. Biotechnology Research Centre, Malaysian Agricultural Research and Development Institute, Kuala Lumpur, Malaysia. uab@mardi.my Biotechnology-derived food crops are currently being developed in Malaysia mainly for disease resistance and improved post harvest quality. The modern biotechnology approach is adopted because of its potential to overcome constraints faced by conventional breeding techniques. Research on the development of biotechnology-derived papaya, pineapple, chili, passion fruit, and citrus is currently under way. Biotechnology-derived papaya developed for resistance to papaya ringspot virus (PRSV) and improved postharvest qualities is at the field evaluation stage. Pineapple developed for resistance to fruit black heart disorder is also being evaluated for proof-of-concept. Other biotechnology-derived food crops are at early stages of gene cloning and transformation. Activities and products involving biotechnology-derived crops will be fully regulated in the near future under the Malaysian Biosafety Law. At present they are governed only by guidelines formulated by the Genetic Modification Advisory Committee (GMAC), Malaysia. Commercialization of biotechnology-derived crops involves steps that require GMAC approval for all field evaluations and food-safety assessments before the products are placed on the market. Public acceptance of the biotechnology product is another important factor for successful commercialization. Understanding of biotechnology is generally low among Malaysians, which may lead to low acceptance of biotechnology-derived products. Initiatives are being taken by local organizations to improve public awareness and acceptance of biotechnology. Future research on plant biotechnology will focus on the development of nutritionally enhanced biotechnology-derived food crops that can provide more benefits to consumers. Publication Types: Review PMID: 16465992 [PubMed - indexed for MEDLINE] 801: Food Nutr Bull. 2005 Dec;26(4):416-8. Improving rice nutrition: challenges and practical approaches for iron fortification. Yoshihara T, Takaiwa F, Goto F. Bio-Science Department, Central Research Institute Electric Power Industry, Abiko, Japan. yoshiha@criepi.denken.or.jp Publication Types: Research Support, Non-U.S. Gov't Review PMID: 16465988 [PubMed - indexed for MEDLINE] 802: Trends Biotechnol. 2006 Mar;24(3):102-4. Epub 2006 Feb 7. Unintended effects in genetically modified crops: revealed by metabolomics? Rischer H, Oksman-Caldentey KM. VTT Technical Research Centre of Finland, VTT Biotechnology, Tietotie 2, Espoo, FIN-02044 VTT, Finland. In Europe the commercialization of food derived from genetically modified plants has been slow because of the complex regulatory process and the concerns of consumers. Risk assessment is focused on potential adverse effects on humans and the environment, which could result from unintended effects of genetic modifications: unintended effects are connected to changes in metabolite levels in the plants. One of the major challenges is how to analyze the overall metabolite composition of GM plants in comparison to conventional cultivars, and one possible solution is offered by metabolomics. The ultimate aim of metabolomics is the identification and quantification of all small molecules in an organism; however, a single method enabling complete metabolome analysis does not exist. Given a comprehensive extraction method, a hierarchical strategy--starting with global fingerprinting and followed by complementary profiling attempts--is the most logical and economic approach to detect unintended effects in GM crops. Publication Types: Review PMID: 16460820 [PubMed - indexed for MEDLINE] 803: J Agric Food Chem. 2006 Feb 8;54(3):682-7. Novel reference gene, PKABA1, used in a duplex real-time polymerase chain reaction for detection and quantitation of wheat- and barley-derived DNA. Rønning SB, Berdal KG, Andersen CB, Holst-Jensen A. Section for Feed and Food Microbiology, National Veterinary Institute, Ullevålsveien 68, Post Office Box 8156 Dep., 0033 Oslo, Norway. We report the development of a duplex real-time Polymerase Chain Reaction (PCR) for the simultaneous detection and quantification of wheat- and barley-derived DNA. We used a single primer pair to amplify the single-copy gene PKABA1 from wheat and barley, using minor-groove-binding probes to distinguish between the two cereals. The assay was fully specific, and different wheat and barley cultivars exhibited similar Ct values, indicating stability across cultivars with respect to allelic and copy number composition. The limits of detection were 5 and 10 PCR-forming units for wheat and barley, respectively, making the duplex assay as sensitive as other singleplex reference gene systems published. We were able to detect both wheat and barley simultaneously in real food samples, and the duplex assay is considered to be suitable as an endogenous reference gene system for the detection and quantification of wheat and barley in genetically modified organisms (GMO) and other food and feed analyses. Publication Types: Research Support, Non-U.S. Gov't PMID: 16448168 [PubMed - indexed for MEDLINE] 804: J Agric Food Chem. 2006 Feb 8;54(3):678-81. Validated method for quantification of genetically modified organisms in samples of maize flour. Kunert R, Gach JS, Vorauer-Uhl K, Engel E, Katinger H. Institute of Applied Microbiology, University of Natural Resources and Applied Life Sciences, Muthgasse 18, A-1190 Vienna, Austria. Renate.Kunert@boku.ac.at Sensitive and accurate testing for trace amounts of biotechnology-derived DNA from plant material is the prerequisite for detection of 1% or 0.5% genetically modified ingredients in food products or raw materials thereof. Compared to ELISA detection of expressed proteins, real-time PCR (RT-PCR) amplification has easier sample preparation and detection limits are lower. Of the different methods of DNA preparation CTAB method with high flexibility in starting material and generation of sufficient DNA with relevant quality was chosen. Previous RT-PCR data generated with the SYBR green detection method showed that the method is highly sensitive to sample matrices and genomic DNA content influencing the interpretation of results. Therefore, this paper describes a real-time DNA quantification based on the TaqMan probe method, indicating high accuracy and sensitivity with detection limits of lower than 18 copies per sample applicable and comparable to highly purified plasmid standards as well as complex matrices of genomic DNA samples. The results were evaluated with ValiData for homology of variance, linearity, accuracy of the standard curve, and standard deviation. Publication Types: Research Support, Non-U.S. Gov't PMID: 16448167 [PubMed - indexed for MEDLINE] 805: Plant Physiol. 2006 Mar;140(3):1047-58. Epub 2006 Jan 27. Cinnamate metabolism in ripening fruit. Characterization of a UDP-glucose:cinnamate glucosyltransferase from strawberry. Lunkenbein S, Bellido M, Aharoni A, Salentijn EM, Kaldenhoff R, Coiner HA, Muñoz-Blanco J, Schwab W. Biomolecular Food Technology, Technical University Munich, Freising, Germany. Strawberry (Fragaria x ananassa) fruit accumulate (hydroxy)cinnamoyl glucose (Glc) esters, which may serve as the biogenetic precursors of diverse secondary metabolites, such as the flavor constituents methyl cinnamate and ethyl cinnamate. Here, we report on the isolation of a cDNA encoding a UDP-Glc:cinnamate glucosyltransferase (Fragaria x ananassa glucosyltransferase 2 [FaGT2]) from ripe strawberry cv Elsanta that catalyzes the formation of 1-O-acyl-Glc esters of cinnamic acid, benzoic acid, and their derivatives in vitro. Quantitative real-time PCR analysis indicated that FaGT2 transcripts accumulate to high levels during strawberry fruit ripening and to lower levels in flowers. The levels in fruits positively correlated with the in planta concentration of cinnamoyl, p-coumaroyl, and caffeoyl Glc. In the leaf, high amounts of Glc esters were detected, but FaGT2 mRNA was not observed. The expression of FaGT2 is negatively regulated by auxin, induced by oxidative stress, and by hydroxycinnamic acids. Although FaGT2 glucosylates a number of aromatic acids in vitro, quantitative analysis in transgenic lines containing an antisense construct of FaGT2 under the control of the constitutive 35S cauliflower mosaic virus promoter demonstrated that the enzyme is only involved in the formation of cinnamoyl Glc and p-coumaroyl Glc during ripening. Publication Types: Research Support, Non-U.S. Gov't PMID: 16443693 [PubMed - indexed for MEDLINE] 806: FEBS Lett. 2006 Feb 6;580(3):955-9. Epub 2006 Jan 19. Metabolic engineering of coenzyme Q by modification of isoprenoid side chain in plant. Takahashi S, Ogiyama Y, Kusano H, Shimada H, Kawamukai M, Kadowaki K. Department of Genetic Diversity, National Institute of Agrobiological Sciences, 2-1-2 Kannondai, Tsukuba, Ibaraki 305-8602, Japan. Coenzyme Q (CoQ), an electron transfer molecule in the respiratory chain and a lipid-soluble antioxidant, is present in almost all organisms. Most cereal crops produce CoQ9, which has nine isoprene units. CoQ10, with 10 isoprene units, is a very popular food supplement. Here, we report the genetic engineering of rice to produce CoQ10 using the gene for decaprenyl diphosphate synthase (DdsA). The production of CoQ9 was almost completely replaced with that of CoQ10, despite the presence of endogenous CoQ9 synthesis. DdsA designed to express at the mitochondria increased accumulation of total CoQ amount in seeds. Publication Types: Research Support, Non-U.S. Gov't PMID: 16442107 [PubMed - indexed for MEDLINE] 807: Bull Entomol Res. 2006 Feb;96(1):43-52. Effects of Galanthus nivalis agglutinin (GNA) expressed in tomato leaves on larvae of the tomato moth Lacanobia oleracea (Lepidoptera: Noctuidae) and the effect of GNA on the development of the endoparasitoid Meteorus gyrator (Hymenoptera: Braconidae). Wakefield ME, Bell HA, Fitches EC, Edwards JP, Gatehouse AM. Central Science Laboratory, Sand Hutton, York, YO41 1LZ, UK. m.wakefield@csl.gov.uk The effect of ingestion of transgenic tomato leaves expressing the plant lectin Galanthus nivalis agglutinin (GNA) on development of larvae of Lacanobia oleracea (Linnaeus) was studied under laboratory conditions. When L. oleracea larvae were fed on tomato line 14.1H, expressing approximately 2.0% GNA, significant increases in the mean larval weight and in the amount of food consumed were found. This resulted in an overall reduction in the mean development time to the pupal stage of approximately 7 days. A significant increase in the percentage survival to the adult moth was also recorded when newly hatched larvae were reared on transgenic tomato leaves (72%) compared to larvae reared on untransformed leaves (40%). The effects of ingestion of GNA by L. oleracea larvae, via artificial diet or the leaves of transgenic tomato or potato plants, on the subsequent development of its solitary endoparasitoid Meteorus gyrator (Thunberg) was also studied. No significant effects on the life cycle parameters of M. gyrator developing in L. oleracea fed on GNA-containing diets were observed. Experiments with transgenic potato plants indicated that the stadium of the host larvae at parasitism had a greater influence on M. gyrator development than the presence of GNA. Potential GNA-binding glycoproteins were detected in the gut and body tissues of larval M. gyrator. Despite detection in host tissues, GNA could not be detected in adult M. gyrator and therefore it is likely that at the time of pupation M. gyrator are able to void the GNA in the meconial pellet. PMID: 16441904 [PubMed - indexed for MEDLINE] 808: Shokuhin Eiseigaku Zasshi. 2005 Dec;46(6):270-6. [Laboratory performance study of the quantitative detection method for genetically modified soybeans (roundup ready soybeans 40-3-2)] [Article in Japanese] Kasama K, Watanabe T, Kikuchi H, Suzuki T, Tokishita S, Sakata K, Matsuki A, Hino A, Akiyama H, Maitani T. Hatano Research Institute, Food and Drug Safety Center, 729-5, Ochiai, Hadano, Kanagawa 257-8523, Japan. To investigate important factors affecting the analytical results, a laboratory-performance study was attempted for the Japanese official methods to detect genetically modified (GM) soybeans (40-3-2). Test samples containing 0, 1 and 5% GM soya powder in non-GM soya powder was prepared. A set of 3 test samples was sent to the participating laboratories along with the protocol. The data were collected from all laboratories and statistically analyzed. In the real-time PCR detection method, the average values of the GM 1% and 5% samples were both much lower than the spiked value because the laboratories using a silica-membrane DNA extraction method underestimated the GM value. On the other hand, the laboratories using other extraction methods, such as the CTAB method obtained values close to the spiked value. These results suggest that use of the silica-membrane DNA extraction method may result in underestimation of the GM content in the real-time PCR method. In the ELISA method, the average value of 5% spiked samples appears to be slightly higher than the fortified value. But, overall, it was considered that reported values were close to the spiked level. Publication Types: English Abstract Research Support, Non-U.S. Gov't PMID: 16440788 [PubMed - indexed for MEDLINE] 809: Clin Exp Allergy. 2006 Feb;36(2):238-48. Evaluation of allergenicity of genetically modified soybean protein extract in a murine model of oral allergen-specific sensitization. Gizzarelli F, Corinti S, Barletta B, Iacovacci P, Brunetto B, Butteroni C, Afferni C, Onori R, Miraglia M, Panzini G, Di Felice G, Tinghino R. Department of Infectious, Parasitic and Immune-mediated Diseases, Istituto Superiore di Sanità, Rome, Italy. BACKGROUND: With the development of genetically modified crop plants there has been a growing interest in the approaches available to assess the potential allergenicity of novel gene products. For additional assessment of the potential allergenicity of expressed proteins, informative data can be generated using animal models. Soybean is one of the major source of protein in human and animal nutrition, and has also been well characterized as a major allergenic source. Advances in biotechnology have resulted in an increasing number of genetically engineered foods, and among these soybean is one of the most widespread. OBJECTIVE: To develop and characterize a murine model of IgE-mediated soybean sensitization induced by intragastric immunization, in the presence of Cholera Toxin, with wild-type soybean extract (wt-SE) or with genetically modified soybean extract (gm-SE). METHODS: Balb/c mice born in our animal facilities, from females fed on soy-free food, were fed with the same soy-free food and used in all the experiments. Mice were sensitized by gavages with soybean extracts, and allergen-specific IgE and IgG responses were studied by direct ELISA and ELISA inhibition. Antigen-specific cell proliferation and cytokine production were evaluated in spleen cell cultures. Results Sensitization with both soybean extracts induced high levels of antigen-specific IgE and IgG1 and low levels of specific IgG2a. Both wt-SE and gm-SE were able to inhibit the binding of specific IgE from mice immunized with gm-SE to the same antigen used for the ELISA coating. A comparable proliferative response was obtained with the homologous as well as with the heterologous extracts. CONCLUSION: In sensitized mice, we observed a predominantly T-helper type 2 (Th2)-type immune response, with increased soybean-specific IgE and IgG1 antibodies and a concomitant increase of IL-4 and IL-5 production. RESULTS: obtained by specific IgE ELISA inhibition and by antigen-specific T cell proliferation demonstrated that wt-SE and gm-SE shared B and T epitopes. The present murine model of soybean sensitization established by the oral route should provide valuable information about risk assessment for food allergy from new proteins of genetically modified foods. PMID: 16433863 [PubMed - indexed for MEDLINE] 810: Therapie. 2005 Sep-Oct;60(5):469-76. [Mechanism of action of antidepressant drugs: importance of genetically modified mice in the pharmacological in vivo approach] [Article in French] Gardier A. Laboratoire de Neuropharmacologie EA3544, Faculté de Pharmacie, Université Paris-Sud, Chatenay-Malabry, France. alain.gardier@cep.u-psud.fr The main hypothesis regarding the mechanism of action of antidepressant drugs is monoaminergic and mainly involves two neurotransmitters, serotonin and noradrenaline. Despite the well-recognized therapeutic efficacy of selective serotonin reuptake inhibitors (SSRIs), some disadvantages still occur. For example, they often require 4-6 weeks to achieve clinical benefits in depressed patients. In the past, some molecules that could shorten this long delay of action have been identified. The role of presynaptic autoreceptors - the activation of which leads to an inhibitory feedback control on neurotransmitter synthesis and release - has been extensively studied for antidepressant effects. In our laboratory, we studied the combined effects of an SSRI and a serotonin autoreceptor antagonist of the 5-HT1B subtype using intracerebral in vivo microdialysis in awake, freely moving mice. Important information on SSRIs has been obtained by applying this technique to genetically modified animals, such as constitutive knockout (KO) mice lacking 5-HT1B receptors (5-HT1B KO) generated by homologous recombination: we compared the effects of a combined treatment on extracellular/intrasynaptic levels of serotonin in various nerve terminals area in wild-type control and KO mice. Thus, we found that indirect activation of 5-HT1B autoreceptors limits the effects of SSRIs on dialysate 5-HT levels at serotonergic nerve terminals such as the ventral hippocampus. The study of substance P (neurokinin 1 receptor [R-NK1]) offers another example of the use of KO mice in the development of a new class of antidepressant drugs. NK1 receptor antagonists may display anxiolytic/antidepressant-like properties. The lack of selective compounds for each tachykinin receptor subtype (R-NK 1, R-NK2 or R-NK3) and differences in their affinity between animal species have made R-NK1 KO mice a very useful experimental tool. In collaborative work we found that genetic (R-NK1 KO mice) or pharmacological (GR205171) blockade of R-NK1 is associated with several changes: the increase in cortical 5-HT outflow caused by systemic injection of paroxetine was 4- to 6-fold higher in freely moving R-NK1 KO mice than in wild-type controls. The constitutive lack of NK1 receptors is associated with a functional desensitization of somatodendritic 5-HT1A autoreceptors, resembling that induced by chronic treatment with SSRI antidepressants. These results highlight the link between a neurotransmitter (serotonin) and a neuropeptide (substance P). This genetic strategy allowed us to point out that multiple targets participate to the effects of classical antidepressant drugs within the brain. We hope that, soon, some mice lines (constitutive or tissue specific, conditional rescue mice having alterations of sleep/wakefulness and/or food intake, altered central serotonin and/or noradrenaline neurotransmission, deficit in neurotrophic factors, but increases in intrasynaptic concentrations of substance P) could be a relevant model of the physiopathology of depressive disorders, and could help us understand the appearance of some symptoms. These recent findings suggest that instead of being rejected, the monoaminergic hypothesis of depression should be improved, corrected and completed by studying the role of other neurotransmitter, neuromodulatory compounds (substance P, BDNF [brain-derived neurotrophic factor]). By doing so, it thus could be possible to improve antidepressant drug treatment, i.e. shorten their long delay of action and/or to decrease treatment resistance or improve its tolerance. Publication Types: English Abstract PMID: 16433012 [PubMed - indexed for MEDLINE] 811: J Dairy Sci. 2006 Feb;89(2):518-24. Production and processing of milk from transgenic goats expressing human lysozyme in the mammary gland. Maga EA, Shoemaker CF, Rowe JD, Bondurant RH, Anderson GB, Murray JD. Department of Animal Science, University of California, Davis 95616, USA. eamaga@ucdavis.edu The potential for applying biotechnology to benefit animal agriculture and food production has long been speculated. The addition of human milk components with intrinsic antimicrobial activity and positive charge to livestock milk by genetic engineering has the potential to benefit animal health, as well as food safety and production. We generated one line of transgenic goats as a model for the dairy cow designed to express human lysozyme in the mammary gland. Here we report the characterization of the milk from 5 transgenic females of this line expressing human lysozyme in their milk at 270 microg/mL or 68% of the level found in human milk. Milk from transgenic animals had a lower somatic cell count, but the overall component composition of the milk and milk production were not different from controls. Milk from transgenic animals had a shorter rennet clotting time and increased curd strength. Milk of such nature may be of benefit to the producer by influencing udder health and milk processing. PMID: 16428620 [PubMed - indexed for MEDLINE] 812: Food Chem Toxicol. 2006 Jul;44(7):964-73. Epub 2006 Jan 19. Sub-chronic (13-week) oral toxicity study in rats with recombinant human lactoferrin produced in the milk of transgenic cows. Appel MJ, van Veen HA, Vietsch H, Salaheddine M, Nuijens JH, Ziere B, de Loos F. TNO Quality of Life, Business Unit Toxicology and Applied Pharmacology, P.O. Box 360, 3700 AJ Zeist, The Netherlands. appel@voeding.tno.nl The oral toxicity of recombinant human lactoferrin (rhLF) produced in the milk of transgenic cows was investigated in Wistar rats by daily administration via oral gavage for 13 consecutive weeks, 7 days per week. The study used four groups of 20 rats/sex/dose. The control group received physiological saline and the three test groups received daily doses of 200, 600 and 2000 mg of rhLF per kg body weight. Clinical observations, growth, food consumption, food conversion efficiency, water consumption, neurobehavioural testing, ophthalmoscopy, haematology, clinical chemistry, renal concentration test, urinalysis, organ weights and gross examination at necropsy and microscopic examination of various organs and tissues were used as criteria for detecting the effects of treatment. Overall, no treatment-related, toxicologically significant changes were observed. The few findings that may be related to the treatment (lower cholesterol in high-dose females, lower urinary pH in high-dose males and females and very slightly higher kidney weight in high-dose females) were considered of no toxicological significance. Based on the absence of treatment-related, toxicologically relevant changes, the no-observed-adverse-effect level (NOAEL) was considered to be at least 2000 mg/kg body weight/day. PMID: 16426723 [PubMed - indexed for MEDLINE] 813: J Plant Physiol. 2005 Dec;162(12):1355-66. Analysis and use of the tobacco eIF4A-10 promoter elements for transgene expression. Tian L, Wu K, Hannam C, Latoszek-Green M, Sibbald S, Hu M, Brown DC, Miki B. Southern Crop Protection and Food Research Centre, Agriculture and Agri-Food Canada, London, Ont, Canada. tianl@agr.gc.ca The eIF4A gene family codes for proteins which unwind secondary structures of mRNA during translational initiation. The tobacco eIF4A-10 promoter is one of a few of constitutive promoters found in plants. Research was conducted to identify the proximal promoter elements and to evaluate the potential application of the promoter for regulating transgene expression in a range of crop plants. A large intron (892 bp) in the leader sequence was found to be dispensable for constitutive promoter activity and did not contribute to the overall performance of the promoter. Deletion analysis showed that the upstream region between -151 bp and -73bp relative to the transcriptional start site was essential for the high level of expression and the constitutive activity. The data indicated that the elements in this region may coordinate and compensate each other for the high levels of promoter expression. The downstream leader sequence also contained a strong quantitative enhancer element that was essential for the full activity of the eIF4A-10 promoter. The eIF-4A10 promoter was found to be active in a wide range of plant species and tissues indicating that it will be useful for the constitutive expression of transgenes in plants. Publication Types: Evaluation Studies PMID: 16425454 [PubMed - indexed for MEDLINE] 814: Neuron. 2006 Jan 19;49(2):285-95. Imaging taste responses in the fly brain reveals a functional map of taste category and behavior. Marella S, Fischler W, Kong P, Asgarian S, Rueckert E, Scott K. Department of Molecular and Cell Biology, Helen Wills Neuroscience Institute, 291 Life Sciences Addition, University of California, Berkeley, Berkeley, California 94720, USA. The sense of taste allows animals to distinguish nutritious and toxic substances and elicits food acceptance or avoidance behaviors. In Drosophila, taste cells that contain the Gr5a receptor are necessary for acceptance behavior, and cells with the Gr66a receptor are necessary for avoidance. To determine the cellular substrates of taste behaviors, we monitored taste cell activity in vivo with the genetically encoded calcium indicator G-CaMP. These studies reveal that Gr5a cells selectively respond to sugars and Gr66a cells to bitter compounds. Flies are attracted to sugars and avoid bitter substances, suggesting that Gr5a cell activity is sufficient to mediate acceptance behavior and that Gr66a cell activation mediates avoidance. As a direct test of this hypothesis, we inducibly activated different taste neurons by expression of an exogenous ligand-gated ion channel and found that cellular activity is sufficient to drive taste behaviors. These studies demonstrate that taste cells are tuned by taste category and are hardwired to taste behaviors. Publication Types: Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't PMID: 16423701 [PubMed - indexed for MEDLINE] 815: Methods. 2006 Feb;38(2):144-9. Mucosal vaccine delivery of antigens tightly bound to an adjuvant particle made from food-grade bacteria. van Roosmalen ML, Kanninga R, El Khattabi M, Neef J, Audouy S, Bosma T, Kuipers A, Post E, Steen A, Kok J, Buist G, Kuipers OP, Robillard G, Leenhouts K. BiOMaDe Technology Foundation, Nijenborgh 4, 9747 AG Groningen, The Netherlands. Mucosal immunization with subunit vaccines requires new types of antigen delivery vehicles and adjuvants for optimal immune responses. We have developed a non-living and non-genetically modified gram-positive bacterial delivery particle (GEM) that has built-in adjuvant activity and a high loading capacity for externally added heterologous antigens that are fused to a high affinity binding domain. This binding domain, the protein anchor (PA), is derived from the Lactococcus lactis AcmA cell-wall hydrolase, and contains three repeats of a LysM-type cell-wall binding motif. Antigens are produced as antigen-PA fusions by recombinant expression systems that secrete the hybrid proteins into the culture growth medium. GEM particles are then used as affinity beads to isolate the antigen-PA fusions from the complex growth media in a one step procedure after removal of the recombinant producer cells. This procedure is also highly suitable for making multivalent vaccines. The resulting vaccines are stable at room temperature, lack recombinant DNA, and mimic pathogens by their bacterial size, surface display of antigens and adjuvant activity of the bacterial components in the GEM particles. The GEM-based vaccines do not require additional adjuvant for eliciting high levels of specific antibodies in mucosal and systemic compartments. PMID: 16414272 [PubMed - indexed for MEDLINE] 816: Regul Toxicol Pharmacol. 2006 Mar;44(2):136-43. Epub 2006 Jan 9. Lack of cross-reactivity between the Bacillus thuringiensis derived protein Cry1F in maize grain and dust mite Der p7 protein with human sera positive for Der p7-IgE. Ladics GS, Bardina L, Cressman RF, Mattsson JL, Sampson HA. DuPont/Pioneer Crop Genetics Regulatory Science and Registration, Wilmington, DE, USA. gregory.s.ladics@usa.dupont.com Cry1F protein, derived from Bacillus thuringiensis, is effective at controlling lepidopteran pests and a synthetic Cry1F transgene was transferred into maize. For the safety assessment of genetically modified food crops, the allergenic potential of the introduced novel trait(s) is evaluated. Because no single parameter is currently predictive of allergic potential, a 'weight of evidence' approach has been proposed. As part of this assessment, the amino acid (aa) sequence of the Cry1F protein was compared to a database of known allergens using recommended criteria. The Cry1F protein did not show significant similarity or a match of eight contiguous identical aa with any allergen. However, a single six contiguous aa match was identified between Cry1F and the Der p7 protein of the dust mite, Dermatophagoides pteronyssinus. To investigate whether Cry1F was cross-reactive with Der p7, sera from 10 dust mite allergic patients containing Der p 7-specific IgE antibody were used to compare IgE-specific binding. No evidence of cross-reactivity was observed between Cry1F and Der p7. This study provides in vitro IgE sera screening data, that when considered in the context of other bioinformatic data [Hileman R.E., Silvanovich, A., Goodman R.E., Rice E.A., Holleschak G., Astwood J.D., Hefle S.L., 2002. Bioinformatic methods for allergenicity assessment using a comprehensive allergen database. Int. Arch. Allergy Immunol. 128, 280-291; Stadler, M.B., Stadler, B.M., 2003. Allergenicity prediction by protein sequence. FASEB J. 17, 1141-1143.], adds further evidence arguing against the use of a six contiguous identical amino acid search to identify potential cross-reactive allergens. Cry1F is heat labile, rapidly hydrolyzed in an in vitro pepsin resistance assay, not glycosylated and not from an allergenic source. Taken together, these data indicate a lack of allergenic concern for Cry1F. PMID: 16406630 [PubMed - indexed for MEDLINE] 817: Fitoterapia. 2006 Feb;77(2):67-82. Epub 2006 Jan 6. Grain legume proteins and nutraceutical properties. Duranti M. Department of AgriFood Molecular Sciences, Università degli Studi di Milano, Italy. marcello.duranti@unimi.it Grain legumes are a valuable source of food proteins. Their exploitation is expected to grow in relation of a growing world's food needs. Moreover, it is currently taking place a reappraisal of the beneficial effects of legume seed dietary intake, which are the basis for various health claims. Proteins and peptides concur to the observed biological activities of legume seeds, but their effect(s) has(ve) not completely been disclosed. Aims of this review are: to report the most relevant putative positive effects of grain legumes on human health and to give an account of the current knowledge on the demonstrated legume seed protein biological activities. Specific effects on the prevention and treatment of various diseases, mostly of which are typical of the affluent countries, are reported. Examples of studies at molecular level aimed at elucidating of the underlying mechanism(s) are given. The prospects on targeted legume protein exploitation in the nutraceutical area, including the biotechnological approaches, are also considered. Publication Types: Review PMID: 16406359 [PubMed - indexed for MEDLINE] 818: Plant Cell Rep. 2006 May;25(5):425-31. Epub 2006 Jan 11. Transformation of Actinidia eriantha: a potential species for functional genomics studies in Actinidia. Wang T, Ran Y, Atkinson RG, Gleave AP, Cohen D. The Horticulture and Food Research Institute of New Zealand Ltd., Private Bag 92169, Auckland, New Zealand. tianchi.wang@hortresearch.co.nz Protocols were developed for regeneration and Agrobacterium-mediated transformation of Actinidia eriantha Benth. A. eriantha has a number of features that make it a useful tool for functional genomics in Actinidia: the vines are relatively small and non-vigorous in nature, flowers form all over the vine including on lower axillary branches and the species flowers prolifically in greenhouse conditions. Flowering and fruiting of transgenic A. eriantha plants was obtained within 2 years of transformation in a containment greenhouse. GUS (beta-glucuronidase) activity indicating stable expression of the uidA gene was observed in leaf, stem, root, petal and fruit tissues. Molecular evidence for incorporation of transgenes into the A. eriantha genome was obtained by PCR and DNA gel blot analysis. Inheritance of transgenic phenotypes was demonstrated in seedling progeny. Functional genomic studies in kiwifruit have been initiated using transgenic A. eriantha plants. Publication Types: Research Support, Non-U.S. Gov't PMID: 16404600 [PubMed - indexed for MEDLINE] 819: Nat Biotechnol. 2006 Jan;24(1):23-5. Erratum in: Nat Biotechnol. 2006 Apr;24(4):467. Comment in: Nat Biotechnol. 2007 Feb;25(2):169-70. European GMO labeling thresholds impractical and unscientific. Weighardt F. Publication Types: Letter PMID: 16404384 [PubMed - indexed for MEDLINE] 820: Nat Biotechnol. 2006 Jan;24(1):2. Genetically modified mush. [No authors listed] Publication Types: Editorial PMID: 16404374 [PubMed - indexed for MEDLINE] 821: Dev Biol. 2006 Feb 15;290(2):482-94. Epub 2006 Jan 3. Transcriptional activation by extradenticle in the Drosophila visceral mesoderm. Stultz BG, Jackson DG, Mortin MA, Yang X, Beachy PA, Hursh DA. Cellular and Tissue Therapy Branch, Center for Biologics Evaluation and Research, Food and Drug Administration, HFM-730, Bldg. 29B, Rm. 1E16, 8800 Rockville Pike, Bethesda, MD 20892, USA. decapentaplegic (dpp) is a direct target of Ultrabithorax (Ubx) in parasegment 7 (PS7) of the embryonic visceral mesoderm. We demonstrate that extradenticle (exd) and homothorax (hth) are also required for dpp expression in this location, as well as in PS3, at the site of the developing gastric caecae. A 420 bp element from dpp contains EXD binding sites necessary for expressing a reporter gene in both these locations. Using a specificity swap, we demonstrate that EXD directly activates this element in vivo. Activation does not require Ubx, demonstrating that EXD can activate transcription independently of homeotic proteins. Restoration is restricted to the domains of endogenous dpp expression, despite ubiquitous expression of altered specificity EXD. We demonstrate that nuclear EXD is more extensively phosphorylated than the cytoplasmic form, suggesting that EXD is a target of signal transduction by protein kinases. Publication Types: Research Support, N.I.H., Extramural Research Support, U.S. Gov't, P.H.S. PMID: 16403493 [PubMed - indexed for MEDLINE] 822: Am J Clin Nutr. 2006 Jan;83(1):59-64. Zinc absorption in Guatemalan schoolchildren fed normal or low-phytate maize. Mazariegos M, Hambidge KM, Krebs NF, Westcott JE, Lei S, Grunwald GK, Campos R, Barahona B, Raboy V, Solomons NW. Center for Studies of Sensory Impairments, Aging, and Metabolism, Guatemala City, Guatemala. manolomazariegos@yahoo.es BACKGROUND: Poor bioavailability of zinc from high-phytate diets is an important contributory factor to zinc deficiency in low-income populations. OBJECTIVE: The objective of this study was to determine the effect of low-phytate maize consumption on zinc absorption. DESIGN: The participants were apparently healthy children from the Central Highlands of Guatemala. Sixty children (20 per group) were randomly assigned to be fed only the low-phytate maize or 1 of 2 control maizes, the isohybrid wild-type maize or a local maize, for a 10-wk period. During the final week, the fractional absorption of zinc for all meals was measured during 1 d with the use of zinc stable isotopes and a dual isotope ratio technique based on urine enrichment data. RESULTS: Mean (+/-SD) phytate intakes for the low-phytate, wild-type, and local maize groups were 1536 +/- 563, 2056 +/- 517, and 2253 +/- 687 mg/d, respectively. Corresponding zinc intakes were 8.6 +/- 2.5, 8.1 +/- 2.0, and 9.7 +/- 2.6 mg/d, and the dietary phytate:zinc molar ratios were 18 +/- 5, 26 +/- 6, and 23 +/- 5. Corresponding fractional absorptions of zinc were 0.32 +/- 0.07, 0.28 +/- 0.07, and 0.29 +/- 0.06. The respective values for total absorbed zinc were 2.72 +/- 0.88, 2.30 +/- 0.96, and 2.78 +/- 1.04 mg/d. No significant differences in either the fractional absorption of zinc or total absorbed zinc were seen between the maize groups. CONCLUSION: Under the conditions of the present study, zinc absorption was not increased by the long-term use of low-phytate maize in children whose major dietary staple is maize. Publication Types: Randomized Controlled Trial Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't PMID: 16400050 [PubMed - indexed for MEDLINE] 823: Clin Exp Allergy. 2005 Dec;35(12):1638-44. A mutant of the major apple allergen, Mal d 1, demonstrating hypo-allergenicity in the target organ by double-blind placebo-controlled food challenge. Bolhaar ST, Zuidmeer L, Ma Y, Ferreira F, Bruijnzeel-Koomen CA, Hoffmann-Sommergruber K, van Ree R, Knulst AC. Department of Dermatology/Allergology, University Medical Center Utrecht, Utrecht, The Netherlands. s.bolhaar@azu.nl BACKGROUND: Allergen-specific immunotherapy for food allergy has been hindered by severe side-effects in the past. Well-characterized hypo-allergenic recombinant food allergens potentially offer a safe solution. OBJECTIVE: To demonstrate hypo-allergenicity of a mutated major food allergen from apple, Mal d 1, in vitro and in vivo. METHODS: A mutant of the major apple allergen, Mal d 1, was obtained by site-directed mutagenesis exchanging five amino acid residues. Fourteen patients with combined birch pollen-related apple allergy were included in the study. Hypo-allergenicity of the mutant rMal d 1 (rMal d 1mut) compared with rMal d 1 was assessed by in vitro methods, i.e. RAST (inhibition), immunoblotting and basophil histamine release (BHR) and in vivo by skin prick test and double-blind placebo-controlled food challenge (DBPCFC). RESULTS: RAST analysis (n = 14) revealed that IgE reactivity to rMal d 1mut was twofold lower than that of the wild-type molecule (95% confidence interval (CI): 1.7-2.4). RAST inhibition (n = 6) showed a 7.8-fold decrease in IgE-binding potency (95% CI: 3.0-12.6). In contrast to this moderate decrease in IgE-binding potency, the biological activity of rMal d 1mut assessed by SPT and BHR decreased 10-200-fold. Hypo-allergenicity was confirmed by DBPCFC (n = 2) with both recombinant molecules. CONCLUSION: A moderate decrease in IgE-binding potency translates into a potent inhibition of biological activity. This is the first study that confirms by DBPCFC that a mutated recombinant major food allergen is clinically hypo-allergenic. This paves the way towards safer immunotherapy for the treatment of food-allergic patients. Publication Types: Randomized Controlled Trial Research Support, Non-U.S. Gov't PMID: 16393331 [PubMed - indexed for MEDLINE] 824: Science. 2005 Dec 23;310(5756):1908-9. Erratum in: Science. 2006 Mar 3;311(5765):1242. Communication. Social values and the governance of science. Gaskell G, Einsiedel E, Hallman W, Priest SH, Jackson J, Olsthoorn J. Methodology Institute, London School of Economics and Political Science, London WC2A 2AE, UK. g.gaskell@lse.ac.uk Publication Types: Research Support, Non-U.S. Gov't PMID: 16373561 [PubMed - indexed for MEDLINE] 825: Int J Hyg Environ Health. 2006 Jan;209(1):81-8. Epub 2005 Sep 29. Detection of genetically modified DNA sequences in milk from the Italian market. Agodi A, Barchitta M, Grillo A, Sciacca S. Department of Biomedical Sciences, University of Catania, Via S. Sofia n. 87, 95123 Catania, Italy. agodia@unict.it The possible transfer and accumulation of novel DNA and/or proteins in food for human consumption derived from animals receiving genetically modified (GM) feed is at present the object of scientific dispute. A number of studies failed to identify GM DNA in milk, meat, or eggs derived from livestock receiving GM feed ingredients. The present study was performed in order to: (i) develop a valid protocol by PCR and multicomponent analysis for the detection of specific DNA sequences in milk, focused on GM maize and GM soybean; (ii) assess the stability of transgenic DNA after pasteurization treatment and (iii) determine the presence of GM DNA sequences in milk samples collected from the Italian market. Results from the screening of 60 samples of 12 different milk brands demonstrated the presence of GM maize sequences in 15 (25%) and of GM soybean sequences in 7 samples (11.7%). Our screening methodology shows a very high sensitivity and the use of an automatic identification of the amplified products increases its specificity and reliability. Moreover, we demonstrated that the pasteurization process is not able to degrade the DNA sequences in spiked milk samples. The detection of GM DNA in milk can be interpreted as an indicator of fecal or airborne contamination, respectively, with feed DNA or feed particles, although an alternative source of contamination, possibly recognizable in the natural environment can be suggested. Further studies, performed on a larger number of milk samples, are needed to understand the likely source of contamination of milk collected from the Italian market. PMID: 16373205 [PubMed - indexed for MEDLINE] 826: Mar Biotechnol (NY). 2006 Mar-Apr;8(2):103-9. Epub 2006 Jan 1. Production of eicosapentaenoic and docosahexaenoic acid-containing oils in transgenic land plants for human and aquaculture nutrition. Robert SS. Food Futures National Research Flagship Division of Marine and Atmospheric Research, Commonwealth Scientific and Industrial Research Organisation (CSIRO), Hobart, Tasmania 7001, Australia. stan.robert@csiro.au A large body of evidence suggests that there is a significant underconsumption of omega-3, long-chain, polyunsaturated fatty acids (LC-PUFAs) and that this is the cause of multiple chronic diseases and developmental aberrations. The scope for increasing omega-3 LC-PUFA consumption from seafood is limited because global wild fisheries are unable to increase their harvests, and aquaculture fisheries currently rely on wild fisheries as a source of LC-PUFAs. Agricultural production of oils is highly efficient and has the potential to be sustainable. The transfer of genes from marine microalgae and other microorganisms into oilseed crops has shown that the production of terrestrial omega-3 LC-PUFA oils is indeed possible. The specifications of these oils or whole seeds for use in human and Atlantic salmon (Salmo salar) aquaculture nutrition are discussed. Publication Types: Review PMID: 16372159 [PubMed - indexed for MEDLINE] 827: Crit Rev Food Sci Nutr. 2005;45(7-8):607-21. Nutraceutical applications of garlic and the intervention of biotechnology. Bhagyalakshmi N, Thimmaraju R, Venkatachalam L, Murthy KN, Sreedhar RV. Plant Cell Biotechnology Department, Central Food Technological Research Institute, Mysore 570 006, India. pcbt@cscftri.ren.nic.in Garlic (Allium sativum L.) is an important and widely cultivated plant with both culinary and medicinal uses stemming from its biological activities, which include antibiotic, anticancer, anti-thrombotic, and lipid-lowering cardiovascular effects. Though such medicinal use of garlic existed for centuries, there was little scientific support for its therapeutic and pharmacological properties. However, there has been a recent upsurge of research on garlic aiming to understand its exact mechanism of action in each case so that garlic and its products may have more judicious future applications. Since garlic is vegetatively propagated, its improvement for desired traits through conventional means is difficult. The intervention of biotechnological methods such as tissue culture and gene transfer protocols developed recently hold great promise for improving this crop. Due to new innovations in instrumentation and processing technologies coupled with more judicious experimental models, better products are foreseen in the market. The objective of this article was to review the recent developments made towards understanding the mechanism by which garlic imparts different therapeutic effects as well as to review what biotechnology can offer to improve this crop and its products. Publication Types: Review PMID: 16371330 [PubMed - indexed for MEDLINE] 828: BMC Biotechnol. 2005 Dec 21;5:32. Open field trial of genetically modified parthenocarpic tomato: seedlessness and fruit quality. Rotino GL, Acciarri N, Sabatini E, Mennella G, Lo Scalzo R, Maestrelli A, Molesini B, Pandolfini T, Scalzo J, Mezzetti B, Spena A. CRA-Istituto Sperimentale per L'Orticoltura Sezione di Montanaso Lombardo, Italy. pinuzzu.rotino@virgilio.it BACKGROUND: Parthenocarpic tomato lines transgenic for the DefH9-RI-iaaM gene have been cultivated under open field conditions to address some aspects of the equivalence of genetically modified (GM) fruit in comparison to controls (non-GM). RESULTS: Under open field cultivation conditions, two tomato lines (UC 82) transgenic for the DefH9-RI-iaaM gene produced parthenocarpic fruits. DefH9-RI-iaaM fruits were either seedless or contained very few seeds. GM fruit quality, with the exception of a higher beta-carotene level, did not show any difference, neither technological (colour, firmness, dry matter, degrees Brix, pH) nor chemical (titratable acidity, organic acids, lycopene, tomatine, total polyphenols and antioxidant capacity - TEAC), when compared to that of fruits from control line. Highly significant differences in quality traits exist between the tomato F1 commercial hybrid Allflesh and the three UC 82 genotypes tested, regardless of whether or not they are GM. Total yield per plant did not differ between GM and parental line UC 82. Fruit number was increased in GM lines, and GM fruit weight was decreased. CONCLUSION: The use in the diet of fruits from a new line or variety introduces much greater changes than the consumption of GM fruits in comparison to its genetic background. Parthenocarpic fruits, produced under open field conditions, contained 10-fold less seeds than control fruits. Thus parthenocarpy caused by DefH9-RI-iaaM gene represents also a tool for mitigating GM seeds dispersal in the environment. PMID: 16371162 [PubMed - indexed for MEDLINE] 829: Biomed Environ Sci. 2005 Oct;18(5):321-5. Preparation of monoclonal antibody against HPT and its application to detecting marker protein in genetically modified rice. Yang LC, Zhang SX, Pi GH, Li YH, Zhu Z, Yang XG. National Institute of Nutrition and Food Safety, Chinese Center for Disease Control and Prevention, Beijing 100050, China. OBJECTIVE: To produce the monoclonal antibodies (mAbs) against hygromycin B phosphotransferase (HPT) and to develop immunoassay based on mAbs for biosafety assessment of HPT in genetically modified rice (GM rice). METHODS: BALB/c mice were immunized with purified recombinant 6His. HPT protein, and the conventional hybridoma technology was used to generate the monoclonal hybridoma cells. ELISA and Western blot were used to analyze the specificity of mAbs recognizing HPT and the cross reaction with other proteins. A double-Ab sandwich ELISA method was established to detect HPT expression level in the sck gene-modified rice plants. RESULTS: Four hybridomas, named F1, D4-2, D4-4, and D4-5, producing the mAbs against HPT were successfully obtained with the titer of ascetic mAbs ranging from 1x10(-4) to 1x10(-5). Identification of subclass showed that all the produced mAbs belonged to IgG1. Western blot showed specific binding reaction between the mAbs to the HPT proteins expressed in the GM rice. A double sandwich ELISA coated with anti-HPT polyclonal antibody was established with mAbs as sandwich antibody, which showed a sensitivity of 30ng/mL and did not crossreact with other proteins. The expression level of HPT in the leaves of sck-transformed lines was detected (80-150ng/mL). But HPT protein in the grain and seed of GM rice could not be detected using this ELISA assay. CONCLUSION: Anti-HPT mAbs prepared herein have a high specificity and can be used for rapid assay of HPT antigen. The expression level of HPT in the GM rice grain and seed is lower than our ELISA detection limit. Publication Types: Research Support, Non-U.S. Gov't PMID: 16370315 [PubMed - indexed for MEDLINE] 830: J Agric Food Chem. 2005 Dec 28;53(26):10239-43. Degradation of endogenous and exogenous genes of roundup-ready soybean during food processing. Chen Y, Wang Y, Ge Y, Xu B. Institute of Food Safety, Chinese Academy of Inspection and Quarantine, Beijing 100025, People's Republic of China. Roundup-Ready soybeans have been genetically modified to resist the effects of the herbicidal glyphosate and have become the most prevalent transgenic crop in the world. In this work, Roundup-Ready soybeans were used as raw material to study the effects of critical processing procedures such as grinding, cooking, blending, homogenization, sterilization, and spray-drying on the length of DNA fragments of an endogenous gene (lectin) and an exogenous gene (epsps) examined in material from three soybean foods of bean curd, soy milk, and soy powder and from samples taken during their processing. The results showed that various processing procedures caused degradations of both the endogenous and exogenous genes to different degrees. In the grinding procedure, endogenous gene DNA was degraded from 1883 to approximately 836 bp, and exogenous gene DNA was degraded from 1512 to approximately 408 bp. In the blending and squeeze-molding procedures, exogenous gene DNA was also degraded from about 408 to 190 bp, but there was no obvious action on the endogenous gene. After the endogenous and exogenous genes had been degraded to some degree, such as 836 and 408 bp, respectively, they were not evidently affected by cooking procedure at 100 degrees C for 15 min. However, the endogenous gene was further considerably degraded from around 836 to 162 bp in the sterilization procedure at 121 degrees C for 30 s. The effect of the homogenization step on endogenous and exogenous genes was similar to that of the cooking procedure. The coagulation procedure, principally a biochemical reaction, did not greatly affect the exogenous gene but did affect endogenous gene, reducing DNA size from about 836 to 407 bp. Furthermore, the spray-drying procedure, a process of physical shearing, high temperature, and sudden high pressure, distinctly caused degradation of both the lectin and epsps genes, rapidly decreasing the sizes from about 836 to 162 bp for the endogenous gene and from about 408 to 190 bp for the exogenous gene. Publication Types: Research Support, Non-U.S. Gov't PMID: 16366721 [PubMed - indexed for MEDLINE] 831: J Agric Food Chem. 2005 Dec 28;53(26):9971-9. Distortion of genetically modified organism quantification in processed foods: influence of particle size compositions and heat-induced DNA degradation. Moreano F, Busch U, Engel KH. Center of Food and Life Sciences, Technical University of Munich, Am Forum 2, 85350 Freising-Weihenstephan, Germany. Milling fractions from conventional and transgenic corn were prepared at laboratory scale and used to study the influence of sample composition and heat-induced DNA degradation on the relative quantification of genetically modified organisms (GMO) in food products. Particle size distributions of the obtained fractions (coarse grits, regular grits, meal, and flour) were characterized using a laser diffraction system. The application of two DNA isolation protocols revealed a strong correlation between the degree of comminution of the milling fractions and the DNA yield in the extracts. Mixtures of milling fractions from conventional and transgenic material (1%) were prepared and analyzed via real-time polymerase chain reaction. Accurate quantification of the adjusted GMO content was only possible in mixtures containing conventional and transgenic material in the form of analogous milling fractions, whereas mixtures of fractions exhibiting different particle size distributions delivered significantly over- and underestimated GMO contents depending on their compositions. The process of heat-induced nucleic acid degradation was followed by applying two established quantitative assays showing differences between the lengths of the recombinant and reference target sequences (A, deltal(A) = -25 bp; B, deltal(B) = +16 bp; values related to the amplicon length of the reference gene). Data obtained by the application of method A resulted in underestimated recoveries of GMO contents in the samples of heat-treated products, reflecting the favored degradation of the longer target sequence used for the detection of the transgene. In contrast, data yielded by the application of method B resulted in increasingly overestimated recoveries of GMO contents. The results show how commonly used food technological processes may lead to distortions in the results of quantitative GMO analyses. PMID: 16366682 [PubMed - indexed for MEDLINE] 832: Regul Toxicol Pharmacol. 2006 Mar;44(2):182-8. Epub 2005 Dec 20. Improved ELISA method for screening human antigen-specific IgE and its application for monitoring specific IgE for novel proteins in genetically modified foods. Takagi K, Teshima R, Nakajima O, Okunuki H, Sawada J. Division of Biochemistry and Immunochemistry, National Institute of Health Sciences, 1-18-1 Kamiyoga, Setagaya-ku, Tokyo 158-8501, Japan. For monitoring the occurrence of IgE antibody specific for novel proteins in genetically modified (GM) foods, ELISA is the most convenient method. The levels of IgE specific for recombinant proteins, phosphinothricin-N-acetyltransferase (PAT), CP4-EPSPS, and Cry9C were determined by ELISA using the sera from patients allergic to known allergens. Ovalbumin (OVA) and OVA-positive patient sera were used as positive control. In the ELISA, 20-fold-diluted sera tested were mostly negative for the specific IgE. However, the PAT-specific, but not CP4-EPSPS- or Cry9C-specific IgE in some patients was apparently higher than that of the healthy volunteers. To clarify the binding specificity of the antibody, we pre-incubated the sera with soluble PAT, but the inhibition was marginal, suggesting that the binding was non-specific. Therefore, we used 1M NaCl as a washing buffer to remove IgE non-specifically bound to the coated PAT. This washing step efficiently decreased non-specific binding. In contrast, OVA-specific IgE binding to OVA-coated plate was not affected by the washing. Finally, in this pilot study significant levels of IgE antibodies specific for the three proteins were not detected in the sera of Japanese food-allergy patients. Publication Types: Research Support, Non-U.S. Gov't PMID: 16364525 [PubMed - indexed for MEDLINE] 833: Pharmacol Ther. 2006 Aug;111(2):374-83. Epub 2005 Dec 20. Genetically modified plants and food hypersensitivity diseases: usage and implications of experimental models for risk assessment. Prescott VE, Hogan SP. Division of Molecular Bioscience, The John Curtin School of Medical Research, Australian National University, Canberra, ACT, Australia. The recent advances in biotechnology in the plant industry have led to increasing crop production and yield that in turn has increased the usage of genetically modified (GM) food in the human food chain. The usage of GM foods for human consumption has raised a number of fundamental questions including the ability of GM foods to elicit potentially harmful immunological responses, including allergic hypersensitivity. To assess the safety of foods derived from GM plants including allergenic potential, the US FDA, Food and Agriculture Organization of the United Nations (FAO)/World Health Organization (WHO), and the EU have developed approaches for evaluation assessment. One assessment approach that has been a very active area of research and debate is the development and usage of animal models to assess the potential allergenicity of GM foods. A number of specific animal models employing rodents, pigs, and dogs have been developed for allergenicity assessment. However, validation of these models is needed and consideration of the criteria for an appropriate animal model for the assessment of allergenicity in GM plants is required. We have recently employed a BALB/c mouse model to assess the potential allergenicity of GM plants. We have been able to demonstrate that this model is able to detect differences in antigenicity and identify aspects of protein post-translational modifications that can alter antigenicity. Furthermore, this model has also enabled us to examine the usage of GM plants as a therapeutic approach for the treatment of allergic diseases. This review discusses the current approaches to assess the allergenic potential of GM food and particularly focusing on the usage of animal models to determine the potential allergenicity of GM foods and gives an overview of our recent findings and implications of these studies. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 16364445 [PubMed - indexed for MEDLINE] 834: Plant Physiol. 2006 Jan;140(1):150-8. Epub 2005 Dec 16. Grapes on steroids. Brassinosteroids are involved in grape berry ripening. Symons GM, Davies C, Shavrukov Y, Dry IB, Reid JB, Thomas MR. School of Plant Science, University of Tasmania, Hobart, Tasmania 7005, Australia. Fruit ripening is a unique plant developmental process with direct implications for our food supply, nutrition, and health. In contrast to climacteric fruit, where ethylene is pivotal, the hormonal control of ripening in nonclimacteric fruit, such as grape (Vitis vinifera), is poorly understood. Brassinosteroids (BRs) are steroidal hormones, essential for normal plant growth and development but not previously implicated in the ripening of nonclimacteric fruit. Here we show that increases in endogenous BR levels, but not indole-3-acetic acid (IAA) or GA levels, are associated with ripening in grapes. Putative grape homologs of genes encoding BR biosynthesis enzymes (BRASSINOSTEROID-6-OXIDASE and DWARF1) and the BR receptor (BRASSINOSTEROID INSENSITIVE 1) were isolated, and the function of the grape BRASSINOSTEROID-6-OXIDASE gene was confirmed by transgenic complementation of the tomato (Lycopersicon esculentum) extreme dwarf (dx/dx) mutant. Expression analysis of these genes during berry development revealed transcript accumulation patterns that were consistent with a dramatic increase in endogenous BR levels observed at the onset of fruit ripening. Furthermore, we show that application of BRs to grape berries significantly promoted ripening, while brassinazole, an inhibitor of BR biosynthesis, significantly delayed fruit ripening. These results provide evidence that changes in endogenous BR levels influence this key developmental process. This may provide a significant insight into the mechanism controlling ripening in grapes, which has direct implications for the logistics of grape production and down-stream processing. Publication Types: Research Support, Non-U.S. Gov't PMID: 16361521 [PubMed - indexed for MEDLINE] 835: J Chromatogr A. 2006 Apr 21;1112(1-2):353-60. Epub 2005 Dec 15. Quinolizidine alkaloids and phomopsins in lupin seeds and lupin containing food. Reinhard H, Rupp H, Sager F, Streule M, Zoller O. Swiss Federal Office of Public Health, Division of Food Science, CH-3003 Bern, Switzerland. hans.reinhard@bag.admin.ch In recent years there has been growing interest in replacing (genetically modified) soya by lupin. Lupin seeds, flours and lupin containing food have been analyzed in order to assess the relevance of a potential health hazard given by mycotoxins and/or naturally occurring alkaloids. Since not all important alkaloids used for quantitation were commercially available, isolation of lupanine, 13alpha-hydroxylupanine and angustifoline from lupin flours of high alkaloid contents was performed. Alkaloids were analyzed by GC-MS/GC-FID in parallel, while the phomopsin mycotoxins were analyzed by ELISA, since chromatographic methods were not sensitive enough and required time-consuming sample cleanup. The analyzed lupin containing foods were free of phomopsins. In foods where lupin was only a minor constituent the alkaloid content was of no concern. However, roasted lupin beans intended as coffee surrogate had alkaloid contents close to the Australian intervention limit of 200 microg/g. PMID: 16359686 [PubMed - indexed for MEDLINE] 836: Peptides. 2006 Jun;27(6):1179-86. Epub 2005 Dec 13. Design of genetically modified soybean proglycinin A1aB1b with multiple copies of bioactive peptide sequences. Prak K, Maruyama Y, Maruyama N, Utsumi S. Laboratory of Food Quality Design and Development, Graduate School of Agriculture, Kyoto University, Uji, Kyoto 611-0011, Japan. The peptide IIAEK derived from beta-lactoglobulin has a hypocholesterolemic activity greater than that of beta-sitosterol. To create food proteins with multiple copies of this valuable peptide sequence, we introduced tandem multimers of the nucleotide sequence encoding the peptide into DNA regions corresponding to the five variable regions of soybean glycinin A1aB1b subunit, and expressed the mutants in Escherichia coli. The expression level and solubility of the five mutants, each containing four IIAEK sequences in each of the variable regions, were compared. Overall, the expression level and solubility of the mutants with four IIAEK sequences in the variable regions IV and V were the best followed by II > III > I. Further, introduction of the fifth IIAEK sequence to the variable region IV did not decrease expression level and solubility. Increasing the number of IIAEK to 7 and 10 slightly decreased expression level, while their solubility decreased to as low as 40 and 1%, respectively. Various mutations were combined to get a mutant containing as many IIAEK sequences as possible. Some of the resulting mutants were expressed in the soluble form. The mutant containing eight IIAEK from the combination of variable regions IV and V (IV-4 + V-4) showed the best balance of the expression level and solubility, followed by the combination of variable regions II and III (II-4 + III-4). The soluble fractions of these mutants were purified by hydrophobic, gel filtration and ion-exchange column chromatography. Yields of IIAEK peptide released by in vitro digestion with trypsin from both mutants were around 80%. This is the first report that a large amount of a physiologically active peptide could be introduced into food protein. Publication Types: Research Support, Non-U.S. Gov't PMID: 16356590 [PubMed - indexed for MEDLINE] 837: Toxicol Sci. 2006 Mar;90(1):252-8. Epub 2005 Dec 7. The value of short amino acid sequence matches for prediction of protein allergenicity. Silvanovich A, Nemeth MA, Song P, Herman R, Tagliani L, Bannon GA. Monsanto Company, Product Characterization Center, Global Regulatory Affairs, St. Louis, Missouri 63167, USA. andre.silvanovich@monsanto.com Typically, genetically engineered crops contain traits encoded by one or a few newly expressed proteins. The allergenicity assessment of newly expressed proteins is an important component in the safety evaluation of genetically engineered plants. One aspect of this assessment involves sequence searches that compare the amino acid sequence of the protein to all known allergens. Analyses are performed to determine the potential for immunologically based cross-reactivity where IgE directed against a known allergen could bind to the protein and elicit a clinical reaction in sensitized individuals. Bioinformatic searches are designed to detect global sequence similarity and short contiguous amino acid sequence identity. It has been suggested that potential allergen cross-reactivity may be predicted by identifying matches as short as six to eight contiguous amino acids between the protein of interest and a known allergen. A series of analyses were performed, and match probabilities were calculated for different size peptides to determine if there was a scientifically justified search window size that identified allergen sequence characteristics. Four probability modeling methods were tested: (1) a mock protein and a mock allergen database, (2) a mock protein and genuine allergen database, (3) a genuine allergen and genuine protein database, and (4) a genuine allergen and genuine protein database combined with a correction for repeating peptides. These analyses indicated that searches for short amino acid sequence matches of eight amino acids or fewer to identify proteins as potential cross-reactive allergens is a product of chance and adds little value to allergy assessments for newly expressed proteins. PMID: 16338955 [PubMed - indexed for MEDLINE] 838: Nat Biotechnol. 2005 Dec;23(12):1475-6. Comment in: Nat Biotechnol. 2007 Jan;25(1):35-6; author reply 36-7. Reassessing the environmental risks of GM crops. Firbank L, Lonsdale M, Poppy G. Publication Types: Letter PMID: 16333281 [PubMed - indexed for MEDLINE] 839: J Agric Food Chem. 2005 Dec 14;53(25):9708-12. Metabolic engineering of plant cells for biotransformation of hesperedin into neohesperidin, a substrate for production of the low-calorie sweetener and flavor enhancer NHDC. Frydman A, Weisshaus O, Huhman DV, Sumner LW, Bar-Peled M, Lewinsohn E, Fluhr R, Gressel J, Eyal Y. Institute of Horticulture, The Volcani Center, ARO, P.O. Box 6, Bet-Dagan 50250, Israel. Neohesperidin dihydrochalcone (NHDC) is a seminatural, safe, low-calorie sweetener, bitterness blocker, and flavor enhancer with unique properties and applications for the food, beverage, pharmaceutical, and animal feed industries. Current production is limited by the availability of the substrate neohesperidin, a flavonoid that accumulates to significant levels only in the inedible bitter citrus species. We propose a process to convert hesperidin, a tasteless flavonoid extracted from orange peels that are abundant byproducts of the vast orange juice industry, into neohesperidin using metabolic engineering and biotransformation via three steps: (i) extraction of hesperidin from orange peels, (ii) hydrolysis of sugar moieties, and (iii) biotransformation of hesperidin hydrolysis products into neohesperidin. We overcame the current technological bottleneck in biotransformation of hesperidin hydrolysis products into neohesperidin using metabolically engineered plant cell cultures expressing a recombinant flavanone-7-O-glucoside-2-O-rhamnosyltransferase. A small-scale production experiment established the feasibility of the proposed process. Publication Types: Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. PMID: 16332119 [PubMed - indexed for MEDLINE] 840: Plant Cell Rep. 2006 Mar;25(3):214-22. Epub 2005 Dec 6. Stable transformation and direct regeneration in Coffea canephora P ex. Fr. by Agrobacterium rhizogenes mediated transformation without hairy-root phenotype. Kumar V, Satyanarayana KV, Sarala Itty S, Indu EP, Giridhar P, Chandrashekar A, Ravishankar GA. Plant Cell Biotechnology Department, Central Food Technological Research Institute, Mysore 570 020, Karnataka, India. A system for genetic transformation of Coffea canephora by co-cultivation with Agrobacterium rhizogenes harbouring a binary vector has been developed. The objective of the present study was the genetic transformation and direct regeneration of transformants through secondary embryos bypassing an intervening hairy root stage. Transformants were obtained with a transformation efficiency up to 3% depending on the medium adjuvant used. A. rhizogenes strain A4 harbouring plasmid pCAMBIA 1301 with an intron uidA reporter and hygromycin phosphotransferase (hptII) marker gene was used for sonication-assisted transformation of Coffea canephora. The use of hygromycin in the secondary embryo induction medium allowed the selection of transgenic secondary embryos having Ri T-DNA along with the T-DNA from the pCAMBIA 1301 binary vector. In addition transgenic secondary embryos devoid of Ri-T-DNA but with stable integration of the T-DNA from the binary vector were obtained. The putative transformants were positive for the expression of the uidA gene. PCR and Southern blot analysis confirmed the independent, transgenic nature of the analysed plants and indicated single and multiple locus integrations. The study clearly demonstrates that A. rhizogenes can be used for delivering transgenes into tree species like Coffea using binary vectors with Agrobacterium tumefaciens T-DNA borders. Publication Types: Research Support, Non-U.S. Gov't PMID: 16331458 [PubMed - indexed for MEDLINE] 841: Cell Metab. 2005 Dec;2(6):411-20. Leptin regulates insulin sensitivity via phosphatidylinositol-3-OH kinase signaling in mediobasal hypothalamic neurons. Morton GJ, Gelling RW, Niswender KD, Morrison CD, Rhodes CJ, Schwartz MW. Department of Medicine, Harborview Medical Center and University of Washington, Seattle, WA 98104, USA. To investigate whether phosphatidylinositol-3 kinase (PI3K) signaling mediates the metabolic effects of hypothalamic leptin action, adenoviral gene therapy was used to direct expression of leptin receptors to the area of the hypothalamic arcuate nucleus (ARC). This intervention markedly improved insulin sensitivity in genetically obese, leptin-receptor-deficient Koletsky (fa(k)/fa(k)) rats via a mechanism that was not dependent on reduced food intake but was attenuated by approximately 44% by third-ventricular infusion of the PI3K inhibitor LY294002. Conversely, ARC-directed expression of a constitutively active mutant of protein kinase B (PKB/Akt, an enzyme activated by PI3K) mimicked the insulin-sensitizing effect of restored hypothalamic leptin signaling in these animals, despite having no effect on food intake or body weight. These findings suggest that hypothalamic leptin signaling is an important determinant of glucose metabolism and that the underlying neuronal mechanism involves PI3K. Publication Types: Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't PMID: 16330326 [PubMed - indexed for MEDLINE] 842: William Mary Law Rev. 2004 Apr;45(5):2167-259. Gaps, inexperience, inconsistencies, and overlaps: crisis in the regulation of genetically modified plants and animals. Mandal GN. Albany Law School, USA. The regulation of genetically modified products pursuant to statutes enacted decades prior to the advent of biotechnology has created a regulatory system that is passive rather than proactive about risks, has difficulty adapting to biotechnology advances, and is highly fractured and inefficient--transgenic plants and animals are governed by at least twelve different statutes and five different agencies or services. The deficiencies resulting from this piecemeal approach to regulation unnecessarily expose society and the environment to adverse risks of biotechnology and introduce numerous inefficiencies into the regulatory system. These risks and inefficiencies include gaps in regulation, duplicative and inconsistent regulation, unnecessary increases in the cost of and delay in the development and commercialization of new biotechnology products. These deficiencies also increase the risk of further unnecessary biotechnology scares, which may cause public overreaction against biotechnology products, preventing the maximization of social welfare. With science and society poised to soar from first-generation biotechnology (focused on crops modified for agricultural benefit), to next-generation developments (including transgenic fish, insects, and livestock, and pharmaceutical-producing and industrial compound-producing plants and animals), it is necessary to establish a comprehensive, efficient, and scientifically rigorous regulatory system. This Article details how to achieve such a result through fixing the deficiencies in, and risks created by, the current regulatory structure. Ignoring many details, the solutions can be summarized in two categories. First, statutory and regulatory gaps that are identified must be closed with new legislation and regulation. Second, regulation of genetically modified products must be shifted from a haphazard model based on statutes not intended to cover biotechnology to a system based upon agency expertise in handling particular types of risks. PMID: 16329219 [PubMed - indexed for MEDLINE] 843: J Toxicol Environ Health A. 2005 Dec 10;68(23-24):2263-76. Multigeneration reproductive and developmental toxicity study of bar gene inserted into genetically modified potato on rats. Rhee GS, Cho DH, Won YH, Seok JH, Kim SS, Kwack SJ, Lee RD, Chae SY, Kim JW, Lee BM, Park KL, Choi KS. Department of Toxicology, National Institute of Toxicological Research, Korea Food and Drug Administration, Seoul. Each specific protein has an individual gene encoding it, and a foreign gene introduced to a plant can be used to synthesize a new protein. The identification of potential reproductive and developmental toxicity from novel proteins produced by genetically modified (GM) crops is a difficult task. A science-based risk assessment is needed in order to use GM crops as a conventional foodstuff. In this study, the specific characteristics of GM food and low-level chronic exposure were examined using a five-generation animal study. In each generation, rats were fed a solid pellet containing 5% GM potato and non-GM potato for 10 wk prior to mating in order to assess the potential reproductive and developmental toxic effects. In the multigeneration animal study, there were no GM potato-related changes in body weight, food consumption, reproductive performance, and organ weight. Polymerase chain reaction (PCR) was carried out using extracted genomic DNA to examine the possibility of gene persistence in the organ tissues after a long-term exposure to low levels of GM feed. In each generation, the gene responsible for bar was not found in any of the reproductive organs of the GM potato-treated male and female rats, and the litter-related indexes did not show any genetically modified organism (GMO)-related changes. The results suggest that genetically modified crops have no adverse effects on the multigeneration reproductive-developmental ability. PMID: 16326439 [PubMed - indexed for MEDLINE] 844: Altern Ther Health Med. 2005 Nov-Dec;11(6):14-7. The world according to Codex. Levy AR. PMID: 16320855 [PubMed - indexed for MEDLINE] 845: Vestn Ross Akad Med Nauk. 2005;(10):30-7. [Principles of production of genetically modified food sources] [Article in Russian] Kirpichnikov MP, Tyshko NV. Methods of genetic engineering have given a powerful impulse to the development of fundamental and applied biology and biotechnology of plants. Methods of genetic plant transformation, such as agrobacterium-mediated and microprojectile bombardment-mediated transformation have been used for a long time. These methods allow production of transgenic plants which express the genes of interest. Dozens of transgenic plants have been obtained by now, and their number is steadily increasing. Publication Types: English Abstract PMID: 16320704 [PubMed - indexed for MEDLINE] 846: J Health Commun. 2005 Dec;10(8):751-67. Gene cuisine or Frankenfood? The theory of reasoned action as an audience segmentation strategy for messages about genetically modified foods. Silk KJ, Weiner J, Parrott RL. Department of Communication, Michigan State University, East Lansing, Michigan 48824, USA. Genetically modified (GM) foods are currently a controversial topic about which the lay public in the United States knows little. Formative research has demonstrated that the lay public is uncertain and concerned about GM foods. This study (N = 858) extends focus group research by using the Theory of Reasoned Action (TRA) to examine attitudes and subjective norms related to GM foods as a theoretical strategy for audience segmentation. A hierarchical cluster analysis revealed four unique audiences based on their attitude and subjective norm toward GM foods (ambivalent-biotech, antibiotech, biotech-normer, and biotech individual). Results are discussed in terms of the theoretical and practical significance for audience segmentation. Publication Types: Research Support, U.S. Gov't, P.H.S. PMID: 16316937 [PubMed - indexed for MEDLINE] 847: Proc Nutr Soc. 2005 Nov;64(4):487-90. Allergenic potential of novel foods. Meredith C. BIBRA International Ltd, Carshalton, Surrey, UK. clivemeredith@btinternet.com Concerns have been expressed that the introduction of novel foods into the diet might lead to the development of new food allergies in consumers. Novel foods can be conveniently divided into GM and non-GM categories. Decision-tree approaches (e.g. International Life Sciences Institute-International Food Biotechnology Council and WHO/FAO) to assess the allergenic potential of GM foods were developed following the discovery, during product development, of the allergenic potential of GM soyabean expressing a gene encoding a storage protein from Brazil nut (Bertolletia excelsa). Within these decision trees considerations include: the source of the transgene; amino acid homology with known allergens; cross-reactivity with IgE from food-allergic individuals; resistance to proteolysis; prediction using animal models of food allergy. Such decision trees are under constant review as new knowledge and improved models emerge, but they provide a useful framework for the assessment of the allergenic potential of GM foods. For novel non-GM foods the assessment of allergenic potential is more subjective; some foods or food ingredients will need no assessment other than a robust protein assay to demonstrate the absence of protein. Where protein is present in the novel non-GM food, hazard and risk assessments need to be made in terms of the quantity of protein that might be consumed, the identity of individual protein components and their relationships to known food allergens. Where necessary, this assessment would extend to serum screening for potential cross-reactivities, skin-prick tests in previously-sensitised individuals and double-blind placebo-controlled food challenges. Publication Types: Review PMID: 16313692 [PubMed - indexed for MEDLINE] 848: Proc Nutr Soc. 2005 Nov;64(4):481-6. GM organisms and the EU regulatory environment: allergenicity as a risk component. Davies HV. Scottish Crop Research Institute, Invergowrie, Dundee, UK. hdavie@scri.ac.uk The European Food Safety Authority, following a request from the European Commission, has published a guidance document for the risk assessment of GM plants and derived food and feed to assist in the implementation of provisions of Regulation (EC) 1829/2003 of the European Parliament and Council on GM food and feed. This regulation has applied since 18 April 2004. In principle, hazard identification and characterisation of GM crops is conducted in four steps: characterisation of the parent crop and any hazards associated with it; characterisation of the transformation process and of the inserted recombinant DNA, including an assessment of the possible production of new fusion proteins or allergens; assessment of the introduced proteins (toxicity, allergenicity) and metabolites; identification of any other targetted and unexpected alterations in the GM crop, including changes in the plant metabolism resulting in compositional changes and assessment of their toxicological, allergenic or nutritional impact. In relation to allergenicity specifically, it is clear that this property of a given protein is not intrinsic and fully predictable but is a biological activity requiring an interaction with individuals with a predisposed genetic background. Allergenicity, therefore, depends on the genetic diversity and variability in atopic human subjects. Given this lack of complete predictability it is necessary to obtain, from several steps in the risk-assessment process, a cumulative body of evidence that minimises any uncertainty about the protein(s) in question. Publication Types: Review PMID: 16313691 [PubMed - indexed for MEDLINE] 849: Proc Nutr Soc. 2005 Nov;64(4):470-4. Industrial dimensions of food allergy. Crevel R. SEAC Toxicology Laboratory, Unilever Research, Sharnbrook, Bedford, UK. rene.crevel@unilever.com Serious attempts to estimate the impact of allergic reactions to foods on public health did not begin until the 1980s. Until about 15 years ago food allergy was considered a minor aspect of food safety. Two developments probably prompted a radical re-appraisal of that situation. The first was the apparently inexorable rise in the prevalence of atopic diseases, of which food allergy forms a part, with its possible consequences highlighted by some well-publicised severe reactions. The second was the growth of genetic modification technology, manifested by the commercialisation of transgenic crops. Each of these developments impacted on the food industry in distinct ways. On the one hand, consumers with food allergies had to be enabled to avoid specific allergens in products formulated with existing ingredients. Food manufacturers therefore had to identify those specific allergens down to trace amounts in all the ingredients forming the product and label or remove them. On the other hand, the introduction of products using ingredients from novel sources required an assessment of the allergenicity of these ingredients as an integral part of safety assurance. The approaches used by the food industry to protect existing consumers who have food allergies and those at potential risk of sensitisation from novel proteins will be illustrated, emphasising how they need to be built into every stage of the life cycle of a product. Publication Types: Review PMID: 16313689 [PubMed - indexed for MEDLINE] 850: Proc Nutr Soc. 2005 Nov;64(4):458-64. The canine model of dietary hypersensitivity. Day MJ. Division of Veterinary Pathology, Infection and Immunity, School of Clinical Veterinary Science, University of Bristol, Langford, UK. m.j.day@bristol.ac.uk IgE-mediated dietary hypersensitivity affects approximately 1% of the canine population. There are no breed associations and < or =50% of the patients are aged <1 year at presentation. The most common causative allergens are beef, chicken, milk, eggs, maize, wheat and soyabean. Affected dogs generally display cutaneous disease and 10-15% of the patients may have concurrent alimentary involvement. Diagnosis is currently based on dietary restriction followed by provocation. Procedures for the detection of serum allergen-specific IgE and IgG antibodies are widely available, but these tests correlate poorly with clinical presentation and dietary testing. Recent studies have demonstrated the allergen specificity of IgE antibodies by immunoblotting and have described blood lymphocyte proliferative responses to food allergens. In addition to investigations of spontaneously-arising dietary hypersensitivity, it has also proved possible to study this disorder experimentally. Small colonies of dogs sensitive to particular dietary proteins have been used to study clinical and serological responses to allergen challenge. Hypersensitivity has been experimentally induced in dogs of an atopic phenotype by repeated subcutaneous injection of alum-adjuvanted dietary allergen during neonatal life. These models have been used to trial a range of modified protein or hydrolysate diets. The dog provides a unique large-animal model for investigation of the immunopathogenesis of human dietary hypersensitivity. The dog is closely related genetically to man and shares environmental disease triggers with man. Spontaneously arising canine dietary hypersensitivity is a good clinical mimic of the human disease, and ability to therapeutically manipulate this adverse response in the dog might lead to benefits for human patients. Publication Types: Review PMID: 16313687 [PubMed - indexed for MEDLINE] 851: J Chromatogr A. 2006 Jan 20;1103(1):118-24. Epub 2005 Nov 23. Determination of flavonoids in cultivated sugarcane leaves, bagasse, juice and in transgenic sugarcane by liquid chromatography-UV detection. Colombo R, Lanças FM, Yariwake JH. Instituto de Química de São Carlos, Universidade de São Paulo, Caixa Postal 780, 13560-970 São Carlos, SP, Brazil. A high-performance liquid chromatography (HPLC) method with photo-diode array (DAD) detection was developed to separate and quantify flavonoids in sugarcane leaves and bagasse (= the crushed sugarcane refuse from juice extraction), and in sugarcane juice. Sugarcane flavonoids consist of a complex mixture of aglycones and glycosides (including flavonolignan glycosides), and the HPLC-UV method herein proposed is suitable for their quantification as total flavonoids. This method was applied to analyze samples of cultivated sugarcane, commercial juice and transgenic sugarcane leaves. Sugarcane leaves proved a promising source of flavonoids: an average of 1.10 mg of total flavonoids/g plant material was found in fresh leaves. Moreover, the flavonoid content of sugarcane juice (0.6 mg/mL) is comparable to other food sources of flavonoids previously reported. Transgenic sugarcane leaves ("Bowman-Birk" and "Kunitz") were compared with non-modified ("control") plant samples using the proposed HPLC-UV method, which indicated that the content of total flavonoids in transgenic plants is different from that in non-modified sugarcane. Publication Types: Research Support, Non-U.S. Gov't Validation Studies PMID: 16310199 [PubMed - indexed for MEDLINE] 852: Shokuhin Eiseigaku Zasshi. 2005 Oct;46(5):J286-8. [Safety assessment of genetically modified foods] [Article in Japanese] Hayakawa T. Pharmaceuticals and Medical Devices Agency, Chiyoda-ku, Tokyo, Japan. PMID: 16305183 [PubMed - indexed for MEDLINE] 853: J Food Prot. 2005 Nov;68(11):2389-94. Temperature and treatment time influence high hydrostatic pressure inactivation of feline calicivirus, a norovirus surrogate. Chen H, Hoover DG, Kingsley DH. Department of Animal & Food Sciences, University of Delaware, Newark, Delaware 19716-2150, USA. Interest in high hydrostatic pressure processing as a nonthermal pasteurization process for foods continues to increase. Feline calicivirus (FCV), a propagable virus that is genetically related to the nonpropagable human noroviruses, was used for detailed evaluation of the high pressure processing parameters necessary for virus inactivation. Pressure inactivation curves of FCV strain KCD in Dulbecco's modified Eagle medium with 10% fetal bovine serum were obtained at 200 and 250 MPa as a function of time at room temperature. Pressure inactivation curves at 200 and 250 MPa also were determined as a function of temperature ranging from --10 to 50 degrees C at treatment times of 4 and 2 min, respectively. Tailing was observed for inactivation as a function of treatment time, indicating that the linear model was not adequate for describing these curves. The two nonlinear models, the log logistic and Weibull functions, consistently produced better fit to inactivation curves than did the linear model. The mean square errors were 0.381 for the log logistic model, 0.425 for the Weibull model, and 1.546 for the linear model. For inactivation as a function of temperature, FCV was most resistant to pressure at 20 degrees C. Temperatures above and below 20 degrees C significantly increased pressure inactivation of FCV. A 4-min treatment of 200 MPa at --10 and 50 degrees C reduced the titer of FCV by 5.0 and 4.0 log units, respectively; whereas at 20 degrees C the same treatment only reduced the titer by 0.3 log units. These novel results point to the potential for using temperatures above and particularly below room temperature to lower the pressure needed to cause the desired level of virus inactivation. Publication Types: Research Support, U.S. Gov't, Non-P.H.S. PMID: 16300078 [PubMed - indexed for MEDLINE] 854: Biomed Pharmacother. 2005 Dec;59(10):531-40. Epub 2005 Oct 21. Biological and biomedical aspects of genetically modified food. Celec P, Kukucková M, Renczésová V, Natarajan S, Pálffy R, Gardlík R, Hodosy J, Behuliak M, Vlková B, Minárik G, Szemes T, Stuchlík S, Turna J. Biomed Research and Publishing Group, Bratislava, Slovakia. petercelec@gmail.com Genetically modified (GM) foods are the product of one of the most progressive fields of science-biotechnology. There are major concerns about GM foods in the public; some of them are reasonable, some of them are not. Biomedical risks of GM foods include problems regarding the potential allergenicity, horizontal gene transfer, but environmental side effects on biodiversity must also be recognized. Numerous methods have been developed to assess the potential risk of every GM food type. Benefits of the first generation of GM foods were oriented towards the production process and companies, the second generation of GM foods offers, on contrary, various advantages and added value for the consumer. This includes improved nutritional composition or even therapeutic effects. Recombinant probiotics and the principle of alternative gene therapy represent the latest approach of using GM organisms for biomedical applications. This article tries to summarize and to explain the problematic topic of GM food. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 16298508 [PubMed - indexed for MEDLINE] 855: Regul Toxicol Pharmacol. 2006 Mar;44(2):125-35. Epub 2005 Nov 18. Statistical analysis used in the nutritional assessment of novel food using the proof of safety. Hothorn LA, Oberdoerfer R. University of Hannover, LG Biostatistics, Herrenhaeuser Str. 2, D-30419 Hannover, Germany. The safety assessment of Novel Food, including GM biotechnology-derived crops, starts with the comparison of the Novel Food with a traditional counterpart that is generally accepted as safe based on a history of human food use. Substantial equivalence is established if no meaningful difference from the conventional counterpart was found, leading to the conclusion that the Novel Food is as safe and nutritious as its traditional counterpart. In general, the non-significance of p value is used for the proof of safety. From a statistical perspective, the problems connected with such an approach are demonstrated, namely that quite different component-specific false negative error rates result. As an alternative, the proof of safety is discussed with the inherently related definition of safety thresholds. Moreover, parametric and non-parametric confidence intervals for the difference and the ratio to control (conventional line) are described in detail. Finally, the treatment of multiple components for a global proof of safety is explained. PMID: 16298467 [PubMed - indexed for MEDLINE] 856: Appetite. 2006 Jan;46(1):67-74. Epub 2005 Nov 17. Implicit attitudes towards genetically modified (GM) foods: a comparison of context-free and context-dependent evaluations. Spence A, Townsend E. RASPH group, School of Psychology, University of Nottingham, University Park, Nottingham NG7 2RD, UK. lpxas@psychology.nottingham.ac.uk Past research on attitudes towards GM food has focused on measuring explicit attitudes. Here we compared implicit attitudes towards GM foods with explicit attitudes towards GM foods. We used the Go No-Go task to investigate context-free implicit evaluations of GM foods and compared these with evaluations made in the context of ordinary and organic foods. Semantic differential scales were used to evaluate explicit attitudes towards GM foods. As expected, explicit attitudes towards GM foods were found to be neutral. However, contrary to our hypotheses, participants were found to hold positive, rather than neutral, implicit attitudes towards GM foods when these were assessed in a context free manner. In addition, neutral implicit attitudes were found when attitudes were assessed in the context of ordinary or organic foods, again contrasting with our hypotheses. These results imply that implicit attitudes towards GM food are more positive than anticipated and may lead to approach behaviour towards such products. Thus, given the choice, consumers are likely to accept GM food although other incentives may be needed if alternative foods are available. Publication Types: Randomized Controlled Trial Research Support, Non-U.S. Gov't PMID: 16298018 [PubMed - indexed for MEDLINE] 857: Risk Anal. 2005 Oct;25(5):1241-52. Who does the public trust? The case of genetically modified food in the United States. Lang JT, Hallman WK. Department of Sociology and the Food Policy Institute, Rutgers University, New Brunswick, NJ 08901, USA. John@Coolclass.com Trust is important for the perception of many types of risk, including those relating to genetically modified (GM) food. Who the public trusts in any given circumstance, however, is not well understood. In this study of public trust regarding GM food, an exploratory factor analysis with Promax rotation reveals public classification of three common institutional types-evaluators, watchdogs, and merchants. The structure of relationships among these stakeholders can act to enable or constrain public support for this new technology. Evaluators-scientists, universities, and medical professionals-are the most trusted. Watchdogs-consumer advocacy organizations, environmental organizations, and media sources-are moderately trusted. Merchants-grocers and grocery stores, industry, and farmers-are least trusted. While the federal government is seen as closest to being an evaluator, it is not highly correlated with any of the factors. The lack of trust in the organizations with the greatest resources and responsibilities for ensuring the safety of GM food should be seen as an important obstacle to the adoption of the technology. Publication Types: Research Support, U.S. Gov't, Non-P.H.S. PMID: 16297228 [PubMed - indexed for MEDLINE] 858: Anal Chem. 2005 Nov 15;77(22):7421-8. Quantitative detection system for maize sample containing combined-trait genetically modified maize. Akiyama H, Watanabe T, Wakabayashi K, Nakade S, Yasui S, Sakata K, Chiba R, Spiegelhalter F, Hino A, Maitani T. National Institute of Health Sciences, Kamiyoga, Setagaya-ku, Tokyo, Japan. akiyama@nihs.go.jp Various countries have established regulations that stipulate the labeling of agricultural commodities, feed, and food products that contain or are made from genetically modified (GM) material or that contain adventitious GM material in amounts that exceed certain threshold levels. While regulations in some countries refer to GM material on a weight per weight (w/w) percentage, the currently applied detection methods do not directly measure the w/w percentage of the GM material. Depending on the particular method and the sample matrix it is applied to, the conversion of analytical results to a w/w percentage is challenging or not possible. The first rapid PCR system for GM maize detection on a single kernel basis has been developed. The equipment for the grinding of individual kernels and a silica membrane-based 96-well DNA extraction kit were both significantly revised and optimized for this particular purpose, respectively. We developed a multiplex real-time PCR method for the rapid quantification of GM DNA sequences in the obtained DNA solutions. In addition, a multiplex qualitative PCR detection method allows for the simultaneous detection of different GM maize traits in each kernel and thereby for identification of individual kernels that contain a combination of two or more GM traits. Especially for grain samples that potentially contain combined-trait GM maize kernels, the proposed methods can deliver informative results in a rapid, precise, and reliable manner. Publication Types: Research Support, Non-U.S. Gov't PMID: 16285695 [PubMed - indexed for MEDLINE] 859: Genewatch. 2005 May-Jun;18(3):3-5, 16. Without a trace: how a lack of U.S. domestic LMO regulations is undermining international control. Bereano P. University of Washington, USA PMID: 16285121 [PubMed - indexed for MEDLINE] 860: Eubios J Asian Int Bioeth. 2000 Jul;10(4):106-13. Attitudes of the public and scientists to biotechnology in Japan at the start of 2000. Ng MA, Takeda C, Watanabe T, Macer D. Institute of Biological Sciences, University of Tsukuba, Tsukuba Science City 305-8572, Japan. This survey on biotechnology and bioethics was carried out on national random samples of the public and scientists in November 2000-January 2000 [sic]throughout Japan, and attendees at the Novartis Life Science Forum held on 29 September 1999 in Tokyo. The sample size was 297, 370, and 74 respectively. While there is a better awareness of GMOs in 2000 compared to 1991; the trend shows an increase in the perceived risks of GMOs followed by growing resistance in Japan. While a majority of persons believed genetic engineering would make life better over the next twenty years (57%), the proportion of respondents who thought genetic engineering would make life worse over the next twenty years doubled from 1997 to 2000 (from 12% to 25%). Respondents were asked whether they had heard about applications in several areas and the order of familiarity (high-low) was: pest-resistant crops, human genes in bacteria, mouse to develop cancer, food and drinks, pigs with human hearts and pre-implantation diagnosis. A divide of opinion can be seen when the results on benefit, risk and moral acceptability of applications of biotechnology by the public are compared to the forum and scientist samples. A significant change in the acceptance of the public occurred in 2000 where only 22% agreed on the moral acceptability of GM food compared to 41% in 1997. In 2000 fewer people said they are willing (20%) to buy genetically modified fruits that taste better compared to 1997 (36%). The results show less public support for use of gene therapy than 1993 and twice as many scientists rejected gene therapy than they did in 1991. When asked who is best placed to regulate modern biotechnology, the respondents were overwhelmingly in favor of international regulatory bodies, such as the United Nations and the World Health Organization (72%), rather than national bodies. The comparison between scientists and public is interesting, however the more enthusiastic sample were participants from the Novaritis Life Science Forum with its mixed occupations. PMID: 16273712 [PubMed - indexed for MEDLINE] 861: Science. 2005 Nov 4;310(5749):847-50. A direct role for dual oxidase in Drosophila gut immunity. Ha EM, Oh CT, Bae YS, Lee WJ. Division of Molecular Life Science and Center for Cell Signaling Research, Ewha Woman's University, Seoul 120-750, South Korea. Because the mucosal epithelia are in constant contact with large numbers of microorganisms, these surfaces must be armed with efficient microbial control systems. Here, we show that the Drosophila nicotinamide adenine dinucleotide phosphate (NADPH) oxidase enzyme, dual oxidase (dDuox), is indispensable for gut antimicrobial activities. Adult flies in which dDuox expression is silenced showed a marked increase in mortality rate even after a minor infection through ingestion of microbe-contaminated food. This could be restored by the specific reintroduction of dDuox, demonstrating that this oxidase generates a unique epithelial oxidative burst that limits microbial proliferation in the gut. Thus, oxidant-mediated antimicrobial responses are not restricted to the phagocytes, but rather are used more broadly, including in mucosal barrier epithelia. Publication Types: In Vitro Research Support, Non-U.S. Gov't PMID: 16272120 [PubMed - indexed for MEDLINE] 862: Appl Environ Microbiol. 2005 Nov;71(11):7075-82. Survival of genetically modified and self-cloned strains of commercial baker's yeast in simulated natural environments: environmental risk assessment. Ando A, Suzuki C, Shima J. National Food Research Institute, 2-1-12 Kannondai, Tsukuba, Ibaraki 305-8642, Japan. Although genetic engineering techniques for baker's yeast might improve the yeast's fermentation characteristics, the lack of scientific data on the survival of such strains in natural environments as well as the effects on human health prevent their commercial use. Disruption of acid trehalase gene (ATH1) improves freeze tolerance, which is a crucial characteristic in frozen-dough baking. In this study, ATH1 disruptants constructed by genetic modification (GM) and self-cloning (SC) techniques were used as models to study such effects because these strains have higher freeze tolerance and are expected to be used commercially. Behavior of the strains in simulated natural environments, namely, in soil and water, was studied by measuring the change in the number of viable cells and in the concentration of DNA that contains ATH1 loci. Measurements were made using a real-time PCR method during 40 days of cultivation. Results showed that the number of viable cells of GM and SC strains decreased in a time-dependent manner and that the decrease rate was nearly equal to or higher than that for wild-type (WT) yeast. For all three strains (SC, GM, and WT) in the two simulated natural environments (water and soil), the DNA remained longer than did viable cells but the decrease patterns of either the DNA or the viable cells of SC and GM strains had tendencies similar to those of the WT strain. In conclusion, disruption of ATH1 by genetic engineering apparently does not promote the survival of viable cells and DNA in natural environments. Publication Types: Research Support, Non-U.S. Gov't PMID: 16269743 [PubMed - indexed for MEDLINE] 863: Plant J. 2005 Nov;44(4):557-68. Analysis of the histone H3 gene family in Arabidopsis and identification of the male-gamete-specific variant AtMGH3. Okada T, Endo M, Singh MB, Bhalla PL. Plant Molecular Biology and Biotechnology laboratory, Australian Research Council Centre of Excellence for Integrative Legume Research, Institute of Land and Food Resources, The University of Melbourne, Parkville, Victoria 3010, Australia. Histones are major components of chromatin, the protein-DNA complex involved in DNA packaging and transcriptional regulation. Histone genes have been extensively investigated at the genome level in animal systems and have been classified as replication dependent, replication independent or tissue specific. However, no such study is available in a plant system. In this paper we report that there are 15 histone H3 genes in the Arabidopsis genome, including five H3.1 genes, three H3.3 genes and five H3.3-like genes. A gene structure analysis revealed that gene duplication causes redundancy of the histone H3 genes. The expression of one of the H3 genes, termed AtMGH3/At1g19890, is cell-specific, being restricted to the generative and sperm cells of Arabidopsis pollen as shown by in situ hybridisation and reporter gene analysis. Thus, we conclude that in Arabidopsis, AtMGH3 is a male-gamete-specific histone H3 gene. A T-DNA insertion line for AtMGH3 revealed decreased expression and ectopic RNA splicing. The T-DNA insertion lines for AtMGH3/At1g19890 and other H3 genes revealed a normal growth phenotype and reproductive fertility. These findings suggest that other H3 genes are likely to compensate for the T-DNA-insertion-induced loss of a single H3 gene because of the high redundancy of these genes in the Arabidopsis genome. These T-DNA mutant lines should be useful for accumulating different H3 gene mutations in a single plant and for studying replication-dependent and replication-independent H3 genes and the specific role of AtMGH3 in chromatin remodelling and transcriptional regulation during development of male gametes. Publication Types: Research Support, Non-U.S. Gov't PMID: 16262706 [PubMed - indexed for MEDLINE] 864: J Biol Chem. 2006 Jan 6;281(1):334-40. Epub 2005 Oct 31. Familial Parkinson mutant alpha-synuclein causes dopamine neuron dysfunction in transgenic Caenorhabditis elegans. Kuwahara T, Koyama A, Gengyo-Ando K, Masuda M, Kowa H, Tsunoda M, Mitani S, Iwatsubo T. Department of Neuropathology and Neuroscience, Graduate School of Pharmaceutical Sciences, University of Tokyo, Hongo Bunkyoku Tokyo, 113-0033 Japan. Mutations in alpha-synuclein gene cause familial form of Parkinson disease, and deposition of wild-type alpha-synuclein as Lewy bodies occurs as a hallmark lesion of sporadic Parkinson disease and dementia with Lewy bodies, implicating alpha-synuclein in the pathogenesis of Parkinson disease and related neurodegenerative diseases. Dopamine neurons in substantia nigra are the major site of neurodegeneration associated with alpha-synuclein deposition in Parkinson disease. Here we establish transgenic Caenorhabditis elegans (TG worms) that overexpresses wild-type or familial Parkinson mutant human alpha-synuclein in dopamine neurons. The TG worms exhibit accumulation of alpha-synuclein in the cell bodies and neurites of dopamine neurons, and EGFP labeling of dendrites is often diminished in TG worms expressing familial Parkinson disease-linked A30P or A53T mutant alpha-synuclein, without overt loss of neuronal cell bodies. Notably, TG worms expressing A30P or A53T mutant alpha-synuclein show failure in modulation of locomotory rate in response to food, which has been attributed to the function of dopamine neurons. This behavioral abnormality was accompanied by a reduction in neuronal dopamine content and was treatable by administration of dopamine. These phenotypes were not seen upon expression of beta-synuclein. The present TG worms exhibit dopamine neuron-specific dysfunction caused by accumulation of alpha-synuclein, which would be relevant to the genetic and compound screenings aiming at the elucidation of pathological cascade and therapeutic strategies for Parkinson disease. Publication Types: Research Support, Non-U.S. Gov't PMID: 16260788 [PubMed - indexed for MEDLINE] 865: Biotechnol Adv. 2006 Mar-Apr;24(2):143-60. Epub 2005 Oct 27. Baculoviruses-- re-emerging biopesticides. Szewczyk B, Hoyos-Carvajal L, Paluszek M, Skrzecz I, Lobo de Souza M. Department of Molecular Virology, Intercollegiate Faculty of Biotechnology of the University of Gdansk and Medical University of Gdansk, Kladki 24, 80-822 GDANSK, Poland. szewczyk@biotech.univ.gda.pl Biological control of agricultural pests has gained importance in recent years due to increased pressure to reduce the use of agrochemicals and their residues in the environment and food. Viruses of a few families are known to infect insects but only those belonging to the highly specialized family Baculoviridae have been used as biopesticides. They are safe to people and wildlife, their specificity is very narrow. Their application as bioinsecticides was limited until recently because of their slow killing action and technical difficulties for in vitro commercial production. Two approaches for the wider application of baculoviruses as biopesticides will be implemented in future. In countries where use of genetically modified organisms is restricted, the improvements will be mainly at the level of diagnostics, in vitro production and changes in biopesticide formulations. In the second approach, the killing activity of baculoviruses may be augmented by genetic modifications of the baculovirus genome with genes of another natural pathogen. It is expected that the baculoviruses improved by genetic modifications will be gradually introduced in countries which have fewer concerns towards genetically modified organisms. Publication Types: Review PMID: 16257169 [PubMed - indexed for MEDLINE] 866: J Comp Neurol. 2005 Dec 5;493(1):63-71. Identifying hypothalamic pathways controlling food intake, body weight, and glucose homeostasis. Elmquist JK, Coppari R, Balthasar N, Ichinose M, Lowell BB. Department of Medicine and Division of Endocrinology, Diabetes, and Metabolism, Beth Israel Deaconess Medical Center, and Harvard Medical School, Boston, Massachusetts 02115, USA. jelmquis@bidmc.harvard.edu The past decade has greatly increased our understanding and appreciation of the ability of the central nervous system (CNS) to regulate food intake and body weight. This was spearheaded by the discovery of key molecules regulating body weight homeostasis. It is now also apparent that the CNS, especially the hypothalamus, plays a primary role in directly regulating glucose homeostasis, independently of effects on body weight. These discoveries are important given the increasing incidences of obesity and type II diabetes in Western societies. In this article, we will highlight recent data from genetically modified mice. These data and other models have helped to dissect the CNS pathways regulating body weight and glucose homeostasis. Finally, although these studies have been illustrative, they also underscore our relative lack of knowledge and highlight the need for more definitive approaches to unravel the functional significance of these pathways. (c) 2005 Wiley-Liss, Inc. Publication Types: Review PMID: 16254991 [PubMed - indexed for MEDLINE] 867: Transgenic Res. 2005 Oct;14(5):775-84. Assessing the transfer of genetically modified DNA from feed to animal tissues. Mazza R, Soave M, Morlacchini M, Piva G, Marocco A. Instituto di Agronomia generale e Coltivazioni erbacee, Università Cattolica S. Cuore, Via E. Parmense, 84, Piacenza 29100, Italy. mazza.raffaele@virgilio.it In Europe, public and scientific concerns about the environmental and food safety of GM (Genetically Modified) crops overshadow the potential benefits offered by crop biotechnology to improve food quality. One of the concerns regarding the use of GM food in human and animal nutrition is the effect that newly introduced sequences may have on the organism. In this paper, we assess the potential transfer of diet-derived DNA to animal tissues after consumption of GM plants. Blood, spleen, liver, kidney and muscle tissues from piglets fed for 35 days with diets containing either GM (MON810) or a conventional maize were investigated for the presence of plant DNA. Only fragments of specific maize genes (Zein, Sh-2) could be detected with different frequencies in all the examined tissues except muscle. A small fragment of the Cry1A(b) transgene was detected in blood, liver, spleen and kidney of the animals raised with the transgenic feed. The intact Cry1A(b) gene or its minimal functional unit were never detected. Statistical analysis of the results showed no difference in recovery of positives for the presence of plant DNA between animals raised with the transgenic feed and animals raised with the conventional feed, indicating that DNA transfer may occur independently from the source and the type of the gene. From the data obtained, we consider it unlikely that the occurrence of genetic transfer associated with GM plants is higher than that from conventional plants. Publication Types: Research Support, Non-U.S. Gov't PMID: 16245168 [PubMed - indexed for MEDLINE] 868: Transgenic Res. 2005 Oct;14(5):761-73. Mammary specific transgenic over-expression of insulin-like growth factor-I (IGF-I) increases pig milk IGF-I and IGF binding proteins, with no effect on milk composition or yield. Monaco MH, Gronlund DE, Bleck GT, Hurley WL, Wheeler MB, Donovan SM. Department of Food Science and Human Nutrition, University of Illinois, Urbana, IL 61801, USA. IGF-I regulates lactation by stimulating mammary mitogenesis, inhibiting apoptosis, and partially mediating the effects of growth hormone on lactogenesis. Herein, lactation performance during first and second parity was assessed in transgenic swine (TG) that over-expressed human IGF-I in milk under the control of the bovine alpha-lactalbumin promoter, regulatory regions and signal peptide coding sequence. Milk samples were collected throughout lactation (farrowing to d24) from TG sows and non-transgenic littermates (CON) and IGF-I, IGF-II, and IGFBP determined. Colostral (<24 h postpartum) IGF-I content was 26-fold greater (p<0.001) in TG sows (949+/- 107 microg/L; range 228-1,600 microg/L) than CON (36+/-17.8 microg/L) and was 50- to 90-fold greater (p< 0.001) in mature milk (d2-24 postpartum). There was no effect of parity on milk IGF-I content. Milk IGF-II concentration was unaffected by IGF-I over-expression. Low molecular weight IGFBP (IGFBP-2 and -5) in the milk of TG sows were higher (p=0.02) than CON in the early postpartum period, but did not differ in mature milk. Milk yield, determined by weigh-suckle-weigh, was similar in TG and CON as was litter weight gain. Milk nutrient composition was not significantly affected by IGF over-expression. Thus, mammary specific transgenic over-expression of IGF-I significantly increased milk IGF-I and IGFBP content, but did not impact lactation performance in swine. Publication Types: Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't PMID: 16245167 [PubMed - indexed for MEDLINE] 869: Transgenic Res. 2005 Oct;14(5):749-59. Crop-to-crop gene flow using farm scale sites of oilseed rape (Brassica napus) in the UK. Weekes R, Deppe C, Allnutt T, Boffey C, Morgan D, Morgan S, Bilton M, Daniels R, Henry C. Central Science Laboratory, YO41 1LZ Sand Hutton, York, UK. From 2000-2003 a range of Farm Scale Evaluation (FSE) trials were established in the UK to assess the effect of the release and management of herbicide tolerant (HT) crops on the abundance and diversity of farmland wildlife compared with their conventionally managed non-GM-equivalents. The objective of this research project was to investigate gene flow within the winter (WOSR) and spring oilseed rape (SOSR) FSE trials and to develop a statistical model for the prediction of cross-pollination frequency that can be used to evaluate current separation distance guidelines. Seed samples were collected from the non-GM half of the trial sites and were tested for evidence of cross-pollination from the GM HT halves using a quantitative PCR assay specific to the HT (bar) gene. Rates of cross-pollination were found to decrease rapidly with increasing distance from the GM source. The quantitative data were subjected to statistical analysis and a two-step model was found to provide the best fit for the data. Significant differences were found between the results for WOSR, SOSR and varietal association (VA) crops. The model predicted that the %GM content (including upper 95% confidence limits) of a sample taken at a distance of 50 m away from the GM source would be 0.04% (0.84%) for WOSR, 0.02% (0.39%) for SOSR, 0.77% (21.72%) for WOSR VA and 0.37% (5.18%) for SOSR VA. The data and models presented here are discussed in the context of necessary separation distances to meet various possible thresholds for adventitious presence of GM in OSR. Publication Types: Research Support, Non-U.S. Gov't PMID: 16245166 [PubMed - indexed for MEDLINE] 870: Vet Res Commun. 2005 Aug;29 Suppl 2:31-4. Role of the "National Reference Centre for Genetically Modified Organisms (GMO) detection" in the official control of food and feed. Ciabatti I, Marchesi U, Froiio A, Paternò A, Ruggeri M, Amaddeo D. Department of Virology and Biotechnology, National Reference Centre for Genetically Modified Organisms (GMO) detection, Istituto Zooprofilattico Sperimentale Lazio e Toscana, via Appia Nuvoa 1411, 00178, Rome, Italy. iciabatti@rm.izs.it The National Reference Centre for Genetically Modified Organisms (GMO) detection was established in 2002 within the Istituto Zooprofilattico Sperimentale Lazio e Toscana, with the aim of providing scientific and technical support to the National Health System and to the Ministry of Health within the scope of the regulation of GMO use in food and feed.The recently adopted EU legislation on GMOs (Regulation CE no. 1829/2003 and no. 1830/2003) introduced more rigorous procedures for the authorisation, labelling and analytical control of food and feed consisting, containing or derived from GMOs. The National Reference Centre, besides its institutional tasks as one of the laboratories of the Italian National Health System, collects and analyses data and results of the national official control of GMOs; carries out scientific research aimed at developing, improving, validating and harmonising detection and quantification methods, in cooperation with other scientific institutions, the Community Reference Laboratory and within the European Network of GMOs laboratories (ENGL); collaborates with the Ministry of Health in the definition of control programmes and promotes educational and training initiatives. Objectives defined for 2004-2006, activities in progress and goals already achieved are presented. Publication Types: Review PMID: 16244921 [PubMed - indexed for MEDLINE] 871: Vet Res Commun. 2005 Aug;29 Suppl 2:19-26. Aspects connected with the enforcement of the EU provisions on genetically modified organisms. Marabelli R. General Directorate for Veterinary Health and Food, Ministry of Health, Rome, Italy. alimentivet@sanita.it Publication Types: Review PMID: 16244919 [PubMed - indexed for MEDLINE] 872: Vet Res Commun. 2005 Aug;29 Suppl 2:7-11. GMO: human health risk assessment. D'Agnolo G. Department of Cell Biology and Neuroscienzes, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161, Rome, Italy. dagnolo@iss.it Publication Types: Review PMID: 16244917 [PubMed - indexed for MEDLINE] 873: Vet Clin North Am Food Anim Pract. 2005 Nov;21(3):637-53, vi. Neonatal care of high-risk cloned and transgenic calves. Fecteau ME, Palmer JE, Wilkins PA. Food Animal Medicine and Surgery, Widener Veterinary Hospital, New Bolton Center, University of Pennsylvania, 382 West Street Road, Kennett Square, PA 19348, USA. mfecteau@vet.upenn.edu Publication Types: Review PMID: 16243628 [PubMed - indexed for MEDLINE] 874: J Appl Microbiol. 2005;99(5):1082-9. Growth of Lactobacillus plantarum in media containing hydrolysates of fish viscera. Horn SJ, Aspmo SI, Eijsink VG. Department of Chemistry, Biotechnology and Food Science, Norwegian University of Life Sciences, As, Norway. svein.horn@umb.no AIMS: To compare growth of Lactobacillus plantarum on media containing hydrolysates (peptones) from cod viscera with growth on commercial media. METHODS AND RESULTS: Growth of Lact. plantarum on various fish peptones and commercial peptones/extracts was evaluated using both a Bioscreen apparatus (microtiter plates, no pH control) and fermentors (with pH control). Generally, the performance of the fish peptones was good and only beaten by the performance of yeast extract. Replacement of the 22 g l(-1) complex nitrogen source in standard MRS medium with only 5 g l(-1) fish peptone reduced the biomass yield with only 10%, whereas replacement with a mixture of 2.5 g l(-1) fish peptone and 2.5 g l(-1) yeast extract increased the biomass yield by 10%. CONCLUSIONS: Peptones derived from cod viscera support excellent growth of Lact. plantarum. SIGNIFICANCE AND IMPACT OF THE STUDY: We show that peptones derived from cod viscera are promising constituents of growth media for fastidious food bacteria such as lactobacilli. Media containing these peptones show excellent performance while problems associated with the use of meat-derived peptones (BSE, kosher status) or plant-derived peptones (genetically modified organisms) are avoided. Publication Types: Research Support, Non-U.S. Gov't PMID: 16238738 [PubMed - indexed for MEDLINE] 875: J Biosci Bioeng. 2002;94(6):536-44. Genetically modified industrial yeast ready for application. Akada R. Department of Applied Chemistry and Chemical Engineering, Faculty of Engineering, Yamaguchi University, Tokiwadai, Ube 755-8611, Japan. rinji@yamaguchi-u.ac.jp Tremendous progress in the genetic engineering of yeast had been achieved at the end of 20th century, including the complete genome sequence, genome-wide gene expression profiling, and whole gene disruption strains. Nevertheless, genetically modified (GM) baking, brewing, wine, and sake yeasts have not, as yet, been used commercially, although numerous industrial recombinant yeasts have been constructed. The recent progress of genetic engineering for the construction of GM yeast is reviewed and possible requirements for their application are discussed. 'Self-cloning' yeast will be the most likely candidate for the first commercial application of GM microorganisms in food and beverage industries. PMID: 16233347 [PubMed] 876: Science. 2005 Oct 14;310(5746):231-3; author reply 231-3. Comment on: Science. 2005 Apr 29;308(5722):688-90. Debate over a GM rice trial in China. Cleveland DA, Soleri D. Publication Types: Comment Letter PMID: 16231407 [PubMed - indexed for MEDLINE] 877: Science. 2005 Oct 14;310(5746):231-3; author reply 231-3. Comment on: Science. 2005 Apr 29;308(5722):688-90. Debate over a GM rice trial in China. Sze PC, Cotter J. Publication Types: Comment Letter PMID: 16231406 [PubMed - indexed for MEDLINE] 878: Int J Cancer. 2006 Apr 1;118(7):1623-7. Daily timed meals dissociate circadian rhythms in hepatoma and healthy host liver. Davidson AJ, Straume M, Block GD, Menaker M. Department of Biology, University of Virginia, Charlottesville, VA 22904, USA. Ad2h@virginia.edu Dividing cells, including human cancers, organize processes necessary for their duplication according to circadian time. Recent evidence has shown that disruption of central regulation of circadian rhythms can increase the rate at which a variety of cancers develop in rodents. To study circadian rhythms in liver tumors, we have chemically induced hepatocellular carcinoma in transgenic rats bearing a luciferase reporter gene attached to the promoter of a core circadian clock gene (Period 1). We explanted normal liver cells and hepatomas, placed them into short-term culture, and precisely measured their molecular clock function by recording light output. Results show that isolated hepatocellular carcinoma is capable of generating circadian rhythms in vitro. Temporally restricting food availability to either day or night altered the phase of the rhythms in both healthy and malignant tissue. However, the hepatomas were much less sensitive to this signal resulting in markedly different phase relationships between host and tumor tissue as a function of mealtime. These data support the conclusion that hepatoma is differentially sensitive to circadian timing signals, although it maintains the circadian organization of the nonmalignant cells from which it arose. Because circadian clocks are known to modulate the sensitivity of many therapeutic cytotoxic targets, controlling meal-timing might be used to increase the efficacy of treatment. Specifically, meal and treatment schedules could be designed that take advantage of coincident times of greatest tumor sensitivity and lowest sensitivity of host tissue to damage. Publication Types: Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't PMID: 16231323 [PubMed - indexed for MEDLINE] 879: Food Addit Contam. 2005 Oct;22(10):1061-71. Risk perception and communication: lessons for the food and food packaging industry. Renn O. Interdisciplinary Research Unit on Risk, Sustainable Technology Development and Governance, University of Stuttgart, Seidenstrasse 36, D-70174 Stuttgart, Germany. Ortwin.renn@soz.uni-stuttgart.de Health risks are front-page news. Be it bovine spongiform encephalitis (BSE), surface ozone, or radiation from transmitter stations or mobile phones, the popular press puts out a constant stream of risk warnings and sensational reports about potential health threats. This paper examines how the general public perceives and assesses such information when it comes to food and food packaging risks. In the first part, the basic components of food risks are discussed and then compared with the perceptions of these risks. The main emphasis is on the risks from food packaging. The term 'perception' as used in cognitive psychology applies to the mental processes through which a person takes in, deals with and assesses information from the environment (physical and communicative) via the senses. The last part of the paper deals with the consequences of risk assessment and risk perception for risk management and risk communication. PMID: 16227190 [PubMed - indexed for MEDLINE] 880: Science. 2005 Oct 14;310(5746):231-3; author reply 231-3. Comment on: Science. 2005 Apr 29;308(5722):688-90. Debate over a GM rice trial in China. Heong KL, Chen YH, Johnson DE, Jahn GC, Hossain M, Hamilton RS. Publication Types: Comment Letter PMID: 16224002 [PubMed - indexed for MEDLINE] 881: Plant Mol Biol. 2005 Sep;59(1):205-19. Modification of endogenous natural genes by gene targeting in rice and other higher plants. Iida S, Terada R. Division of Molecular Genetics, National Institutes of Natural Sciences, National Institute for Basic Biology, Myodaiji, Okazaki 444-8585, Japan. shigiida@nibb.ac.jp The capability to modify a genomic sequence into a designed sequence is a powerful tool for biologists and breeders to elucidate the function of an individual gene and its cis-acting elements of multigene families in the genome. Gene targeting refers to the alteration of a specific DNA sequence in an endogenous gene at its original locus in the genome. In higher plants, however, the overwhelming occurrence of the random integration of transgenes by non-homologous end-joining is the main obstacle to develop efficient gene targeting. Two approaches have been undertaken to modify a genomic sequence in higher plants- chimeric RNA/DNA oligonucleotide-directed gene targeting to generate a site-specific base conversion, and homologous recombination-dependent gene targeting to produce either a base change or a gene replacement in a sequence-specific manner. The successful and reproducible targeting of an endogenous gene by homologous recombination, independently of gene-specific selection by employing a strong positive-negative selection, has been demonstrated for the first time in rice, an important staple food and a model plant for other cereal species. This review addresses the current status of targeting of an endogenous natural gene in rice and other higher plants and discusses possible models for Agrobacterium- mediated gene targeting by homologous recombination using a strong positive-negative selection. Publication Types: Research Support, Non-U.S. Gov't PMID: 16217613 [PubMed - indexed for MEDLINE] 882: Eur J Histochem. 2005 Jul-Sep;49(3):237-42. Reversibility of hepatocyte nuclear modifications in mice fed on genetically modified soybean. Malatesta M, Tiberi C, Baldelli B, Battistelli S, Manuali E, Biggiogera M. Istituto di Istologia e Analisi di Laboratorio, University of Urbino Carlo Bo, via Zeppi s.n., 61029 Urbino, Italy. malatesta@uniurb.it In the literature, the reports on the effects of a genetically modified (GM) diet are scanty and heterogeneous; in particular, no direct evidence has so far been reported that GM food may affect human or animal health. Hepatocytes represent a suitable model for monitoring the effects of a GM diet, the liver potentially being a primary target. In a previous study, we demonstrated that some modifications occur in hepatocyte nuclei of mice fed on GM soybean. In order to elucidate whether such modifications can be reversed, in the present study, 3 months old mice fed on GM soybean since their weaning were submitted to a diet containing wild type soybean, for one month. In parallel, to investigate the influence of GM soybean on adult individuals, mice fed on wild type soybean were changed to a GM diet, for the same time. Using immunoelectron microscopy, we demonstrated that a one-month diet reversion can influence some nuclear features in adult mice, restoring typical characteristics of controls in GM-fed animals, and inducing in control mice modifications similar to those observed in animals fed on GM soybean from weaning. This suggests that the modifications related to GM soybean are potentially reversible, but also that some modifications are inducible in adult organisms in a short time. Publication Types: Comparative Study Research Support, Non-U.S. Gov't PMID: 16216809 [PubMed - indexed for MEDLINE] 883: Biotechnol Annu Rev. 2005;11:335-54. Detection of metazoan species as a public health issue: simple methods for the validation of food safety and quality. Vassioukovitch O, Orsini M, Paparini A, Gianfranceschi G, Cattarini O, Di Michele P, Montuori E, Vanini GC, Romano Spica V. University of Movement Sciences (IUSM), Section of Hygiene, Department, Human Movement and Sport Sciences, P.zza L. De Bosis 6, 00194 Foro Italico-Rome, Italy. Species identification represents a critical issue in food chain safety and quality control. Several procedures are available to detect animal proteins in cattle feed or to trace transgenic foods. The most effective approach is based on the use of DNA as a marker. Amplification of DNA provides rapid, sensitive and specific protocols. Several target genes can be used, but new insights come from the mitochondrial genome, which is naturally amplified in each cell and shows a remarkable resistance to degradation. These are key points when analysing complex matrices such as foods, animal feedstuff or environmental samples. Traceability is important to prevent BSE or to monitor novel foods, such as genetically modified organisms. Amplification is commonly performed, but it requires expertise and a molecular biology laboratory to perform restriction analysis, electrophoresis or gel staining for the visualisation of results. Hereby, we consider a strategy based on multiple nested amplification and reverse hybridisation assay that virtually requires only a thermocycler and a water bath. The protocol is rapid and simple and can simultaneously detect different species in a DNA sample. This promising approach allows microarray developments, opening up to further perspectives. An international application has been published under the patent cooperation treaty. Presently, a ban on feeding ruminants on cattle-derived proteins is in force in Europe and USA. The identification of metazoan traces in a sample is not only a mere preventive measure for BSE, but represents a possible screening system for monitoring biotechnology products and procedures, as well as a quality control strategy to assure consumer's rights. Publication Types: Review PMID: 16216782 [PubMed - indexed for MEDLINE] 884: Appl Microbiol Biotechnol. 2005 Nov;69(2):126-32. Epub 2005 Nov 12. Microbial fibrinolytic enzymes: an overview of source, production, properties, and thrombolytic activity in vivo. Peng Y, Yang X, Zhang Y. College of Life Sciences, Sichuan University, Chengdu, PR China. Accumulation of fibrin in the blood vessels usually results in thrombosis, leading to myocardial infarction and other cardiovascular diseases. For thrombolytic therapy, microbial fibrinolytic enzymes have now attracted much more attention than typical thrombolytic agents because of the expensive prices and the undesirable side effects of the latter. The fibrinolytic enzymes were successively discovered from different microorganisms, the most important among which is the genus Bacillus from traditional fermented foods. The physiochemical properties of these enzymes have been characterized, and their effectiveness in thrombolysis in vivo has been further identified. Therefore, microbial fibrinolytic enzymes, especially those from food-grade microorganisms, have the potential to be developed as functional food additives and drugs to prevent or cure thrombosis and other related diseases. Publication Types: Review PMID: 16211381 [PubMed - indexed for MEDLINE] 885: Environ Biosafety Res. 2005 Jan-Mar;4(1):13-27. A conceptual framework for the design of environmental post-market monitoring of genetically modified plants. Sanvido O, Widmer F, Winzeler M, Bigler F. Agroscope FAL Reckenholz, Swiss Federal Research Station for Agroecology and Agriculture, CH-8046 Zurich, Switzerland. olivier.sanvido@fal.admin.ch Genetically modified plants (GMPs) may soon be cultivated commercially in several member countries of the European Union (EU). According to EU Directive 2001/18/EC, post-market monitoring (PMM) for commercial GMP cultivation must be implemented, in order to detect and prevent adverse effects on human health and the environment. However, no general PMM strategies for GMP cultivation have been established so far. We present a conceptual framework for the design of environmental PMM for GMP cultivation based on current EU legislation and common risk analysis procedures. We have established a comprehensive structure of the GMP approval process, consisting of pre-market risk assessment (PMRA) as well as PMM. Both programs can be distinguished conceptually due to principles inherent to risk analysis procedures. The design of PMM programs should take into account the knowledge gained during approval for commercialization of a specific GMP and the decisions made in the environmental risk assessments (ERAs). PMM is composed of case-specific monitoring (CSM) and general surveillance. CSM focuses on anticipated effects of a specific GMP. Selection of case-specific indicators for detection of ecological exposure and effects, as well as definition of effect sizes, are important for CSM. General surveillance is designed to detect unanticipated effects on general safeguard subjects, such as natural resources, which must not be adversely affected by human activities like GMP cultivation. We have identified clear conceptual differences between CSM and general surveillance, and propose to adopt separate frameworks when developing either of the two programs. Common to both programs is the need to put a value on possible ecological effects of GMP cultivation. The structure of PMM presented here will be of assistance to industry, researchers, and regulators, when assessing GMPs during commercialization. Publication Types: Research Support, Non-U.S. Gov't PMID: 16209133 [PubMed - indexed for MEDLINE] 886: Transgenic Res. 2005 Aug;14(4):449-62. Risk analysis for plant-made vaccines. Kirk DD, McIntosh K, Walmsley AM, Peterson RK. The Biodesign Institute and School of Life Sciences, Arizona State University, Tempe, AZ 85287, USA. dwayne.kirk@asu.edu The production of vaccines in transgenic plants was first proposed in 1990 however no product has yet reached commercialization. There are several risks during the production and delivery stages of this technology, with potential impact on the environment and on human health. Risks to the environment include gene transfer and exposure to antigens or selectable marker proteins. Risks to human health include oral tolerance, allergenicity, inconsistent dosage, worker exposure and unintended exposure to antigens or selectable marker proteins in the food chain. These risks are controllable through appropriate regulatory measures at all stages of production and distribution of a potential plant-made vaccine. Successful use of this technology is highly dependant on stewardship and active risk management by the developers of this technology, and through quality standards for production, which will be set by regulatory agencies. Regulatory agencies can also negatively affect the future viability of this technology by requiring that all risks must be controlled, or by applying conventional regulations which are overly cumbersome for a plant production and oral delivery system. The value of new or replacement vaccines produced in plant cells and delivered orally must be considered alongside the probability and severity of potential risks in their production and use, and the cost of not deploying this technology--the risk of continuing with the status quo alternative. Publication Types: Research Support, Non-U.S. Gov't PMID: 16201411 [PubMed - indexed for MEDLINE] 887: Regul Toxicol Pharmacol. 2006 Feb;44(1):43-8. Epub 2005 Sep 26. Science, politics, and the GM debate in Europe. Tencalla F. Monsanto Europe S.A., Avenue de Tervuren 270-272, B-1150 Brussels, Belgium. francesca.tencalla@monsanto.com Europe today stands at a crossroad, facing challenges but also opportunities. In its intent to make Europe a leading technology-based economy by 2010, the European Commission has identified biotechnology and genomics as fields for future growth, crucial for supporting the agricultural and food processing industry. Since first commercialization in 1996, GM crop areas have grown at double-digit rates, making this one of the most rapidly adopted technologies in agriculture. However, in contrast to other world areas and despite European Commission support, Europe has found itself 'bogged-down' in a polemic between opponents and supporters of plant biotechnology. As a result, planted areas have remained small. This stalemate is due to a lack of political leadership, especially at the Member State level, all the more surprising in light of European early development and competitive advantage with crop biotechnology. This situation proves once again that, for cutting-edge innovations, a solid science base alone is not sufficient. Acceptance or rejection of new technologies depends on interlinked political, economic, and societal factors that create a favorable or unfavorable situation at a given time. This article will look at GM crops in Europe and the role science and politics have played in the introduction of crop biotechnology. PMID: 16188360 [PubMed - indexed for MEDLINE] 888: J Biosci. 2005 Sep;30(4):515-48. Will transgenic plants adversely affect the environment? Velkov VV, Medvinsky AB, Sokolov MS, Marchenko AI. Institute of Biochemistry and Physiology of Microorganisms, Russian Academy of Sciences,Pushchino, Moscow Region, 142290, Russian Federation. vvvelkov@rambler.ru Transgenic insecticidal plants based on Bacillus thuringiensis (Bt) endotoxins, on proteinase inhibitors and on lectins, and transgenic herbicide tolerant plants are widely used in modern agriculture. The results of the studies on likelihood and non-likelihood of adverse effects of transgenic plants on the environment including: (i) effects on nontarget species; (ii) invasiveness; (iii) potential for transgenes to 'escape' into the environment by horizontal gene transfer; and (iv) adverse effects on soil biota are reviewed. In general, it seems that large-scale implementation of transgenic insecticidal and herbicide tolerant plants do not display considerable negative effects on the environments and, moreover, at least some transgenic plants can improve the corresponding environments and human health because their production considerably reduces the load of chemical insecticides and herbicides. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 16184014 [PubMed - indexed for MEDLINE] 889: Shokuhin Eiseigaku Zasshi. 2005 Aug;46(4):J223-7. [Polymerase chain reaction technology for detection of unapproved genetically modified maize line (Bt10 line)] [Article in Japanese] Watanabe T. National Institute of Health Sciences, 1-18-1, Kamiyoga, Setagaya-ku, Tokyo 158-8501, Japan. Publication Types: Review PMID: 16180689 [PubMed - indexed for MEDLINE] 890: New Sci. 2005 Jun 11-17;186(2503):21. Making the world better? Bruce D. Science, Religion and Technology Project, Church of Scotland, UK. PMID: 16178099 [PubMed - indexed for MEDLINE] 891: Acta Biochim Pol. 2005;52(3):725-9. Nutritional properties of tubers of conventionally bred and transgenic lines of potato resistant to necrotic strain of Potato virus Y (PVYN). Juśkiewicz J, Zduńczyk Z, Fornal J. Institute of Animal Reproduction and Food Research of the Polish Academy of Sciences, Olsztyn, Poland. glebczo@pan.olsztyn.pl The potential effect of genetic modification on nutritional properties of potatoes transformed to improve resistance to a necrotic strain of Potato virus Y was determined in a rat experiment. Autoclaved tubers from four transgenic lines were included to a diet in the amount of 40% and compared with the conventional cv. Irga. The experiment lasted 3 weeks and special attention was paid to nutritional properties of diets, caecal metabolism and serum indices. Genetic modification of potato had no negative effect on the chemical composition and nutritional properties of tubers, ecosystem of the caecum, activity of serum enzymes and non-specific defence mechanism of the rats. Obtained results indicate that transgenic potato with improved resistance to PVY(N): line R1F (truncated gene coding for PVY(N) polymerase in sense orientation), R2P (truncated gene coding for PVY(N) polymerase in antisense orientation), and NTR1.16 (non-translated regions of PVY(N) genome in sense orientation) are substantial and nutritional equivalence to the non-transgenic cultivar. Tubers of transgenic line NTR2.27 (non-translated regions of PVY(N) genome in antisense orientation) increased the bulk of caecal digesta and the production of SCFA as compared to tubers of the conventional cultivar and the other transgenic clones. Taking into account some deviations, it seems reasonable to undertake a long-term feeding study to confirm the nutritional properties of tubers of transgenic lines. PMID: 16175247 [PubMed - indexed for MEDLINE] 892: Acta Biochim Pol. 2005;52(3):673-8. Biotechnology of temperate fruit trees and grapevines. Laimer M, Mendonça D, Maghuly F, Marzban G, Leopold S, Khan M, Balla I, Katinger H. Plant Biotechnology Unit, IAM, Deptartment of Biotechnology, BOKU, Vienna, Austria. m.laimer@iam.boku.ac.at Challenges concerning fruit trees and grapevines as long lived woody perennial crops require adapted biotechnological approaches, if solutions are to be found within a reasonable time frame. These challenges are represented by the need for correct identification of genetic resources, with the foreseen use either in conservation or in breeding programmes. Molecular markers provide most accurate information and will be the major solution for questions about plant breeders rights. Providing healthy planting material and rapid detection of newly introduced pathogens by reliable methods involving serological and molecular biological tools will be a future challenge of increases importance, given the fact that plant material travels freely in the entire European Union. But also new breeding goals and transgenic solutions are part of the biotechnological benefits, e.g. resistance against biotic and abiotic stress factors, modified growth habits, modified nutritional properties and altered processing and storage qualities. The successful characterization of transgenic grapevines and stone fruit trees carrying genes of viral origin in different vectors constructed under ecological consideration, will be presented. Beyond technical feasibility, efficiency of resistance, environmental safety and Intellectual Property Rights, also public acceptance needs consideration and has been addressed in a specific project. The molecular determination of internal quality parameters of food can also be addressed by the use of biotechnological tools. Patient independent detection tools for apple allergens have been developed and should allow to compare fruits from different production systems, sites, and genotypes for their content of health threatening compounds. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 16175241 [PubMed - indexed for MEDLINE] 893: Can J Microbiol. 2005 Jul;51(7):591-8. Biological control of apple blue mold with Pseudomonas fluorescens. Etebarian HR, Sholberg PL, Eastwell KC, Sayler RJ. Pacific Agri-Food Research Centre, Agriculture and Agri-Food Canada, Canada. Pseudomonas fluorescens isolate 1100-6 was evaluated as a potential biological control agent for apple blue mold caused by Penicillium expansum or Penicillium solitum. Both the wild-type isolate 1100-6 and a genetically modified derivative labeled with the gene encoding the green fluorescent protein (GFP) were compared. The P. fluorescens isolates with or without GFP equally reduced the growth of Penicillium spp. and produced large zones of inhibition in dual culture plate assays. Cell-free metabolites produced by the bacterial antagonists reduced the colony area of Penicillium isolates by 17.3% to 78.5%. The effect of iron chelate on the antagonistic potential of P. fluorescens was also studied. The use of iron chelate did not have a major effect on the antagonistic activity of P. fluorescens. With or without GFP, P. fluorescens significantly reduced the severity and incidence of apple decay by 2 P. expansum isolates after 11 d at 20 degrees C and by P. expansum and P. solitum after 25 d at 5 degrees C when the biocontrol agents were applied in wounds 24 or 48 h before challenging with Penicillium spp. Populations of P. fluorescens labeled with the GFP were determined 1, 9, 14, and 20 d after inoculation at 5 degrees C. The log CFU/mL per wound increased from 6.95 at the time of inoculation to 9.12 CFU/mL (P < 0.05) 25 d after inoculation at 5 degrees C. The GFP strain did not appear to penetrate deeply into wounds based on digital photographs taken with an inverted fluorescence microscope. These results indicate that P. fluorescens isolate 1100-6 could be an important new biological control for apple blue mold. Publication Types: Evaluation Studies Research Support, Non-U.S. Gov't PMID: 16175208 [PubMed - indexed for MEDLINE] 894: Nat Neurosci. 2005 Oct;8(10):1350-5. Epub 2005 Sep 18. Regulation of aversion to noxious food by Drosophila neuropeptide Y- and insulin-like systems. Wu Q, Zhao Z, Shen P. Department of Cellular Biology and Biomedical and Health Sciences Institute, University of Georgia, 724 Biological Sciences Building, Athens, Georgia 30602, USA. Omnivores, including humans, have an inborn tendency to avoid noxious or unfamiliar foods. Such defensive foraging behaviors are modifiable, however, in response to physiological needs. Here we describe a method for assessing risk-sensitive food acquisition in Drosophila melanogaster. Food-deprived fly larvae become more likely to feed on noxious foods (adulterated with quinine) as the duration of deprivation increases. The neuropeptide F receptor NPFR1, a mammalian neuropeptide Y (NPY) receptor homolog, centrally regulates the response to noxious food in D. melanogaster. Overexpression of NPFR1 was sufficient to cause nondeprived larvae to more readily take in noxious food, whereas loss of NPFR1 signaling led to the opposite phenotype. Moreover, NPFR1 neuronal activity may be directly regulated by the insulin-like signaling pathway. Upregulation of insulin-like receptor signaling in NPFR1 cells suppressed the feeding response to noxious food. Our results suggest that the coordinated activities of the conserved NPY- and insulin-like receptor signaling systems are essential for the dynamic regulation of noxious food intake according to the animal's energy state. Publication Types: Comparative Study Research Support, N.I.H., Extramural Research Support, U.S. Gov't, P.H.S. PMID: 16172603 [PubMed - indexed for MEDLINE] 895: Res Microbiol. 2005 Sep;156(8):837-42. Epub 2005 Jun 22. Genomic fingerprinting of bacteriocin-producer strains of Staphylococcus aureus. Nascimento Jdos S, Giambiagi-deMarval M, de Oliveira SS, Ceotto H, dos Santos KR, Bastos Mdo C. Departamento de Microbiologia Geral, Instituto de Microbiologia Prof. Paulo de Góes, Universidade Federal do Rio de Janeiro, Brazil. Among 363 strains of Staphylococcus aureus, 21 were shown to produce bacteriocins (Bac), antimicrobial peptides with potential biotechnological applications. This collection includes strains which are either isolated from food, patients and healthy cattle, or are involved in subclinical bovine mastitis. From these 21 strains, 17 were shown to carry closely-related 8.0-kb Bac plasmids encoding bacteriocins either identical to or similar to aureocin A70, a bacteriocin able to inhibit strains of Listeria monocytogenes, a food-borne pathogen. Such findings prompted us to investigate the genetic relationships among these Bac+ strains. To obtain more discriminatory results, a combined analysis of AP-PCR, rep-PCR, and a modified PCR technique that we designated SD-PCR was employed. The 17 Bac+ strains harboring 8.0-kb Bac plasmids exhibited seven fingerprint patterns. One such genotype was composed of 8 out of the 11 strains associated with bovine mastitis, which suggests the prevalence of a clone of Bac+ strains involved in this animal infection carrying 8.0-kb Bac plasmids. Our data support the assumption that Bac+ strains of S. aureus carrying genetically related 8.0-kb Bac plasmids do not belong to a single clone. It seems, therefore, that 8.0-kb Bac plasmids have spread horizontally among different S. aureus strains. There also seems to be genetic diversity among the remaining Bac+ strains analyzed. Publication Types: Research Support, Non-U.S. Gov't PMID: 16171981 [PubMed - indexed for MEDLINE] 896: Anal Biochem. 2005 Nov 1;346(1):90-100. Epub 2005 Aug 25. Multiplex polymerase chain reaction and ligation detection reaction/universal array technology for the traceability of genetically modified organisms in foods. Peano C, Bordoni R, Gulli M, Mezzelani A, Samson MC, Bellis GD, Marmiroli N. Institute of Biomedical Technologies, National Research Council, Via Fratelli Cervi 93, Segrate, Milano 20090, Italy. clelia.peano@itb.cnr.it A multiplex polymerase chain reaction (PCR) system was developed for the simultaneous detection of target sequences in genetically modified soybean (Roundup Ready) and maize (MON810, Bt176, Bt11, and GA21). Primer pairs were designed to amplify the junction regions of the transgenic constructs analyzed and the endogenous genes of soybean (lectin) and maize (zein) were included as internal control targets to assess the efficiency of all reactions. This multiplex PCR has constituted the basis for an efficient platform for genetically modified organism traceability based on microarray technology. In particular, the ligation detection reaction combined to a universal array approach, using the multiplex PCR as target, was applied. High specificity and sensitivity were obtained. Publication Types: Research Support, Non-U.S. Gov't PMID: 16169511 [PubMed - indexed for MEDLINE] 897: Plant J. 2005 Oct;44(1):62-75. Metabolic engineering of proanthocyanidins by ectopic expression of transcription factors in Arabidopsis thaliana. Sharma SB, Dixon RA. Plant Biology Division, Samuel Roberts Noble Foundation, 2510 Sam Noble Parkway, Ardmore, OK 73401, USA. Genetic transformation of Arabidopsis thaliana with the Arabidopsis TT2 MYB transcription factor resulted in ectopic expression of the BANYULS gene, encoding anthocyanidin reductase, AHA10 encoding a P-type proton-pump and TT12 encoding a transporter involved in proanthocyanidin biosynthesis. When coupled with constitutive expression of PAP1, a positive regulator of anthocyanin biosynthesis, TT2 expression in Arabidopsis led to accumulation of proanthocyanidins, but only in a subset of cells in which the BANYULS promoter is naturally expressed. Ectopic expression of the maize Lc MYC transcription factor weakly induced AHA10 but did not induce BANYULS, TT12 or accumulation of proanthocyanidins. However, high-level combined expression of TT2, PAP1 and Lc resulted in proanthocyanidin synthesis throughout young leaves and cotyledons, followed by death of the plants 1 to 2 weeks after germination. We discuss these results in relation to engineering proanthocyanidins to improve the quality of food and forage plants. Publication Types: Research Support, Non-U.S. Gov't PMID: 16167896 [PubMed - indexed for MEDLINE] 898: J Anim Sci. 2005 Oct;83(10):2404-13. Effect of a short-term fast on intestinal disaccharidase activity and villus morphology of piglets suckling insulin-like growth factor-I transgenic sows. Hartke JL, Monaco MH, Wheeler MB, Donovan SM. Division of Nutritional Sciences, Beckman Institute for Advanced Science and Technology, University of Illinois, Urbana, 61801, USA. The objectives of this study were to use transgenic sows that overexpress IGF-I in milk to investigate the effect of a short-term fast on piglet intestinal morphology and disaccharidase activity and to determine how milk-borne IGF-I influences the response to fasting. After farrowing, litters were normalized to 10 piglets. On d 6, piglets (n = 30) suckling IGF-I transgenic (TG) sows and piglets (n = 30) suckling nontransgenic sows (control) were assigned randomly to three treatments: fed piglets (0 h), which remained with the sow until euthanized on d 7, or fasted piglets, which were removed from the sow at either 6 or 12 h before euthanasia on d 7. Serum IGF-I and IGFBP, intestinal weight and length, jejunal protein and DNA content, disaccharidase activity, and villus morphology were measured. Fasting for 12 h resulted in a negative weight change between d 6 and 7 (quadratic response to fasting; P < 0.001). Piglets suckling TG sows tended to have greater intestinal length (P = 0.068), but no effect of IGF-I overexpression was noted for intestinal weight. Fasting, however, resulted in linear (P < 0.001) and quadratic (P = 0.002) decreases in intestinal weight. Serum IGF-I did not differ between control and TG sows, but decreased linearly (P = 0.003) with fasting. Serum IGFBP-4 decreased (linear and quadratic; P < or = 0.02) with fasting, whereas IGFBP-1 increased quadratically (P < 0.001) with fasting. Jejunal villus height, width, and crypt depth were all increased with fasting (linear and quadratic; P < 0.04). Disaccharidase activity was not affected by fed state; however, piglets suckling TG sows had greater jejunal lactase-phlorhizin hydrolase (P < 0.01) and sucrase-isomaltase (P = 0.02) activities than control piglets. In summary, intestinal weight, villus morphology, serum IGF-I, serum IGFBP-1 and -4, and piglet BW change were altered (P < or = 0.02) in response to fasting. Thus, the duration of food deprivation before euthanization should be considered when designing experiments to assess intestinal development or the IGF axis, as the magnitude of differences between the fed and fasted state may exceed those expected as a result of experimental treatment. Publication Types: Comparative Study Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. PMID: 16160053 [PubMed - indexed for MEDLINE] 899: J Agric Food Chem. 2005 Sep 21;53(19):7512-7. Genetic manipulation of proline accumulation influences the concentrations of other amino acids in soybean subjected to simultaneous drought and heat stress. Simon-Sarkadi L, Kocsy G, Várhegyi A, Galiba G, de Ronde JA. Department of Biochemistry and Food Technology, Budapest University of Technology and Economics, Hungary. sarkadi@mail.bme.hu The effect of simultaneous drought and heat stress on free amino acid levels was compared in wild type and transgenic soybean (Glycine max (L.) Merr cv Ibis) plants transformed with the cDNA coding for the last enzyme of Pro biosynthesis, l-Delta(1)-pyrroline-5-carboxylate reductase (EC 1.5.1.2), in sense and antisense directions. The most rapid increase in Pro content was found in the sense transformants that exhibited the least water loss, while the slowest elevation of Pro levels was detected in the antisense transformants that exhibited the greatest water loss during stress. Correspondingly, the level of the Pro precursors Glu and Arg was higher in sense transformants and lower in antisense ones compared to the wild type plants during the initial part of the stress. Interestingly, genetic manipulation of Pro levels also affected the stress-induced changes in the concentration of several other amino acids, which indicates the coordinated regulation of their metabolic pathways. Publication Types: Research Support, Non-U.S. Gov't PMID: 16159180 [PubMed - indexed for MEDLINE] 900: Appetite. 2005 Dec;45(3):242-9. Epub 2005 Sep 12. Unpacking atitudes towards genetically modified food. de Liver Y, van der Pligt J, Wigboldus D. Social Psychology Program, University of Amsterdam, Roetersstraat 15, 1018 WB Amsterdam, The Netherlands. j.n.deliver@uva.nl The present study investigates the structure of attitudes towards genetically modified (GM) food. A total of 431 respondents completed a questionnaire measuring their overall attitude, cognition and affect towards GM food. A model with distinct positive and negative, affective and cognitive components and a separate factor for perceived risk and worry best accounted for the data. Negative--but not positive--components directly affected behavioural intentions. Implications of these findings for our understanding of attitudes towards GM food and their impact on behaviour are discussed. Publication Types: Research Support, Non-U.S. Gov't PMID: 16154663 [PubMed - indexed for MEDLINE] 901: Evolution. 2005 Jul;59(7):1560-9. Selection on increased intrinsic growth rates in coho salmon, Oncorhynchus kisutch. Sundström LF, Lõhmus M, Devlin RH. Fisheries and Oceans Canada, 4160 Marine Drive, West Vancouver, British Columbia V7V 1N6, Canada. sundstromf@pac.dfo-mpo.gc.ca Substantial evidence from the animal kingdom shows that there is a trade-off between benefits and costs associated with rapid somatic growth. One would therefore expect growth rates under natural conditions to be close to an evolutionary optimum. Nevertheless, natural selection in many salmonid species appears to be toward larger size and earlier emergence from spawning redds, indicating a potential for increased growth rate to evolve. We tested how selection for genetic variants (growth hormone transgenic coho salmon, Oncorhynchus kisutch, with more than doubled daily growth rate potential relative to wild genotypes) depended on predator timing and food abundance during the early period of life (fry stage). In artificial redds, fry of the fast-growing genotypes showed a highly significant developmental shift, emerging from gravel nests approximately two weeks sooner, but with an 18.6% reduced survival, relative to wild-genotype fry. In seminatural streams, fry of the fast-growing genotypes suffered higher predation than those of wild genotypes when predators were present at the time of fry emergence, but this difference was less pronounced when food was scarce. In streams where predators were introduced after emergence, fry survived equally well regardless of food availability. Surviving fry grew faster in habitats provided with more food, and fast-growing genotypes also grew faster than wild genotypes when predators arrived late and food was abundant. Fewer fish migrated downstream past a waterfall when food availability was high and in the presence of predators, and wild-genotype fry were more likely to migrate than fry of the fast-growing genotypes. After being returned to the experimental streams after migration, fast-growing genotypes survived equally well as those of the same genotypes that did not migrate, whereas migrating wild genotypes experienced higher mortality relative to those of the same genotypes that did not migrate. Comparisons of growth rates between siblings retained under hatchery conditions and those from habitats with the fastest growth in the experimental stream revealed that growth rates were similar for wild genotypes in both environments, whereas the fast-growing genotypes in the streams only realized 90% of their growth potential. The present study has shown that a major shift in developmental timing can alter critical early stages affecting survival and can have a significant effect on fitness. Furthermore, ecological conditions such as food abundance and predation pressure can strongly influence the potential for fast-growing variants to survive under natural conditions. The large-scale removal of many predatory species around the world may augment the evolution of increased intrinsic growth rates in some taxa. Publication Types: Comparative Study Research Support, Non-U.S. Gov't PMID: 16153041 [PubMed - indexed for MEDLINE] 902: Pest Manag Sci. 2005 Dec;61(12):1186-92. Beetle-specific Bacillus thuringiensis Cry3Aa toxin reduces larval growth and curbs reproduction in Spodoptera littoralis (Boisd.). Hussein HM, Habustová O, Sehnal F. Institute of Entomology, Academy of Sciences, Branisovská 31, 37005 Ceské Budejovice, Czech Republic. Application of Bacillus thuringiensis tenebrionis (Bt) and expression of the Bt protein Cry3Aa in genetically modified crops are used for targeted control of the Colorado potato beetle Leptinotarsa decemlineata (Say). The Cry3A proteins are selectively toxic for the beetles but the present study describes effects of Cry3Aa on the Egyptian armyworm, Spodoptera littoralis (Boisduval). Cry3Aa expressed in potatoes or added to an agar-base wheat-germ diet reduced the growth of S. littoralis caterpillars and the fertility of adults. The effect of 1.4 mg kg(-1) Cry3Aa in potato leaves was comparable with that of 3.3 mg kg(-1) in the diet. This difference in activity was correlated with better digestibility and higher conversion efficiency of the diet that also supported higher reproduction rate: S. littoralis grown on the potatoes reached a similar size to those on the diet but laid only 702 instead of 1077 eggs per female. Cry3Aa consumption reduced body growth as a consequence of lower food intake without significantly affecting food digestibility and the conversion efficiency of nutrients. The 11% and 5% body weight reductions caused by 1.4 mg kg(-1) Cry3Aa in potatoes and 3.3 mg kg(-1) in the diet, respectively, were associated with 74% and 65% reduction in the number of progeny; S littoralis grown on a diet with 9.1 mg kg(-1) Cry3Aa were 10% smaller and produced no viable progeny. These data suggest that the curtailment of reproduction was not caused by a general shortage of nutrient reserves but by a more direct Cry3Aa effect on the reproduction process. Publication Types: Research Support, Non-U.S. Gov't PMID: 16152673 [PubMed - indexed for MEDLINE] 903: World J Gastroenterol. 2005 Sep 14;11(34):5381-4. Studies on BN rats model to determine the potential allergenicity of proteins from genetically modified foods. Jia XD, Li N, Wu YN, Yang XG. National Institute for Nutrition and Food Safety, Chinese Center for Disease Control and Prevention, 29 Nanwei Road, Beijing 100050, China. AIM: To develop a Brown Norway (BN) rat model to determine the potential allergenicity of novel proteins in genetically modified food. METHODS: The allergenicity of different proteins were compared, including ovalbumin (OVA), a potent respiratory and food allergen, bovine serum albumin (BSA), a protein that is considered to have a lesser allergenic potential, and potato acid phosphatase (PAP), a non-allergenic protein when administered to BN rats via different routes of exposure (intraperitoneally or by gavage). IgG and IgE antibody responses were determined by ELISA and PCA, respectively. An immunoassay kit was used to determine the plasma histamine level. In addition, possible systemic effect of allergens was investigated by monitoring blood pressure. RESULTS: OVA provoked very vigorous protein-specific IgG and IgE responses, low grade protein-specific IgG and IgE responses were elicited by BSA, while by neither route did PAP elicit anything. In either routes of exposure, plasma histamine level in BN rats sensitized with OVA was higher than that of BSA or PAP. In addition, an oral challenge with BSA and PAP did not induce any effect on blood pressure, while a temporary drop in systolic blood pressure in few animals of each routes of exposure was found by an oral challenge with OVA. CONCLUSION: BN rat model might be a useful and predictive animal model to study the potential allergenicity of novel food proteins. Publication Types: Research Support, Non-U.S. Gov't PMID: 16149151 [PubMed - indexed for MEDLINE] 904: Transgenic Res. 2005 Jun;14(3):261-72. Two different Bacillus thuringiensis toxin genes confer resistance to beet armyworm (Spodoptera exigua Hübner) in transgenic Bt-shallots (Allium cepa L.). Zheng SJ, Henken B, de Maagd RA, Purwito A, Krens FA, Kik C. Plant Research International, Wageningen University and Research Center, P O Box 16, 6700 AA Wageningen, The Netherlands. Agrobacterium-mediated genetic transformation was applied to produce beet armyworm (Spodoptera exigua Hübner) resistant tropical shallots (Allium cepa L. group Aggregatum). A cry1Ca or a H04 hybrid gene from Bacillus thuringiensis, driven by the chrysanthemum ribulose-1,5-bisphosphate carboxylase/oxygenase small subunit (Rubisco SSU) promoter, along with the hygromycin phosphotransferase gene (hpt) driven by the CaMV 35S promoter, was employed for genetic transformation. An average transformation frequency of 3.68% was obtained from two shallot cultivars, Tropix and Kuning. After transfer of the in vitro plants to the greenhouse 69% of the cry1Ca and 39% of the H04 transgenic shallots survived the first half year. After one year of cultivation in the greenhouse the remaining cry1Ca and H04 transgenic plants grew vigorously and had a normal bulb formation, although the cry1Ca transgenic plants (and controls) had darker green leaves compared to their H04 counterparts. Standard PCR, adaptor ligation PCR and Southern analyses confirmed the integration of T-DNA into the shallot genome. Northern blot and ELISA analyses revealed expression of the cry1Ca or H04 gene in the transgenic plants. The amount of Cry1Ca expressed in transgenic plants was higher than the expression levels of H04 (0.39 vs. 0.16% of the total soluble leaf proteins, respectively). There was a good correlation between protein expression and beet armyworm resistance. Cry1Ca or H04 gene expression of at least 0.22 or 0.08% of the total soluble protein in shallot leaves was sufficient to give a complete resistance against beet armyworm. This confirms earlier observations that the H04 toxin is more toxic to S. exigua than the Cry1Ca toxin. The results from this study suggest that the cry1Ca and H04 transgenic shallots developed could be used for introducing resistance to beet armyworm in (sub) tropical shallot. Publication Types: Research Support, Non-U.S. Gov't PMID: 16145834 [PubMed - indexed for MEDLINE] 905: Transgenic Res. 2005 Jun;14(3):237-49. Process development and economic evaluation of recombinant human lactoferrin expressed in rice grain. Nandi S, Yalda D, Lu S, Nikolov Z, Misaki R, Fujiyama K, Huang N. Ventria Bioscience, 4110 North Freeway, Sacramento, CA 95834, USA. In this paper, we show that recombinant human lactoferrin (rhLF) has been stably expressed at 0.5% brown rice flour weight for nine generations. Process development indicates that rhLF can be efficiently extracted from rice flour in 20 mM phosphate buffer (pH 7.0) containing up to 0.5 M NaCl and at a ratio of 1 kg flour to 10 L buffer. After solid/liquid separation, the extract can then be loaded directly onto an ion-exchange column and rhLF can be eluted using 0.8 M NaCl. The resulting rhLF is about 95% pure. A range of biochemical and biophysical analyses were carried out and results indicated that the purified rhLF was identical to its native human counterpart other than its glycosylation. Economic analysis shows that at 600 kg/year scale, the cash cost to produce 1 g of rhLF of pharmaceutical grade is US$ 5.90. Analysis also indicates that the expression level has profound impact on costs related to planting, milling, extraction and purification, thus high level expression of recombinant protein in plants is one of the key parameters for the success of plant made pharmaceuticals. PMID: 16145832 [PubMed - indexed for MEDLINE] 906: Science. 2005 Sep 2;309(5740):1471. European politics. Germany poised to elect first scientist-chancellor. Vogel G. Publication Types: News PMID: 16141035 [PubMed - indexed for MEDLINE] 907: Ann Chim. 2005 Jun;95(6):405-14. A proteomic approach to study protein variation in GM durum wheat in relation to technological properties of semolina. Di Luccia A, Lamacchia C, Fares C, Padalino L, Mamone G, La Gatta B, Gambacorta G, Faccia M, Di Fonzo N, La Notte E. Dipartimento di Produzioni Animali, Università di Bari, via Amendola 156/A, Bari, Italy. Genetic manipulation of durum wheats by tobacco rab-1 genes influence the trafficking of gluten proteins through the secretory system by up- or down-regulating the transport step from the ER to the Golgi apparatus which may in turn modify functional performance of the grain. Gluten proteins were extracted from two genetically manipulated lines - Svevo B730 1-1 and Ofanto B688 1-2 - and their control lines and were analyzed by two dimensional gel electrophoresis. When the two-dimensional maps were compared by image analysis no significant differences between the GM line with an up-regulated trafficking containing the wild type tobacco rab1 (Svevo B730 1-1) and its control (Svevo control). By contrast, significant differences were found between the GM line with a down-regulated trafficking due to the tobacco rab1 mutant form (Ofanto B688 1-2) and its control (Ofanto control). Of the new protein spots detected in the down-regulated Ofanto B688 1-2 map, only a beta-amylase was identified. The remaining spots were susceptible to chymotripsin action but not to trypsin one, as in the case of the gluten protein. Rheological measurements showed that gluten quality was enhanced in the down-regulated Ofanto B688 1-2 without an increase in the amount of gluten. Proteomics is a useful and powerful tool for investigating protein changes in GMOs and in understanding events in food science and technology. PMID: 16136835 [PubMed - indexed for MEDLINE] 908: Physiol Genomics. 2005 Nov 17;23(3):311-7. Epub 2005 Aug 30. Growth, metabolism, and blood pressure disturbances during aging in transgenic rats with altered brain renin-angiotensin systems. Kasper SO, Carter CS, Ferrario CM, Ganten D, Ferder LF, Sonntag WE, Gallagher PE, Diz DI. Hypertension and Vascular Disease Center, Physiology and Pharmacology Department, Wake Forest University School of Medicine, Winston-Salem, North Carolina 27157-1032, USA. Transgenic rats with targeted decreased glial expression of angiotensinogen (ASrAogen rats) did not show an increase in systolic pressure compared with Sprague-Dawley (SD) rats during aging (15-69 wk of age). ASrAogen animals had lower body weights throughout the study, similar to reports for animals with systemic knockout of angiotensinogen or treated long term with renin-angiotensin system (RAS) blockers. Further characterization of indexes of growth and metabolism in ASrAogen rats compared with (mRen2)27 and SD rats, which express elevated versus normal brain and tissue angiotensin II levels, respectively, revealed that serum leptin was 100-200% higher in SD and (mRen2)27 rats at 46 wk and 69 wk of age. Consistent with low serum leptin, ASrAogen rats had higher food intake (73%) compared with SD or (mRen2)27 rats. (mRen2)27 rats had higher resting insulin levels than ASrAogen rats at all ages. Insulin levels were constant during aging in ASrAogen rats, whereas an increase occurred in SD rats, leading to higher insulin levels at 46 and 69 wk of age compared with ASrAogen rats. IGF-1 was comparable among strains at all ages, but (mRen2)27 rats had longer and ASrAogen rats had shorter tail lengths versus SD rats at 15 wk of age. In conclusion, reduced expression of glial angiotensinogen blunts the age-dependent rise in insulin levels and weight gain, findings that mimic the effects of long-term systemic blockade of the RAS or systemic knockout of angiotensinogen. These data implicate glial angiotensinogen in the regulation of body metabolism as well as hormonal mechanisms regulating blood pressure. Publication Types: Comparative Study Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't PMID: 16131528 [PubMed - indexed for MEDLINE] 909: J Agric Food Chem. 2005 Sep 7;53(18):7003-9. Erratum in: J Agric Food Chem. 2006 May 31;54(11):4076. Real-time polymerase chain reaction based assays for quantitative detection of barley, rice, sunflower, and wheat. Hernández M, Esteve T, Pla M. Molecular Genetics Department, Institut de Biologia Molecular de Barcelona (IBMB), Consejo Superior de Investigaciones Científicas, Jordi Girona Salgado 18-26, 08034 Barcelona, Spain. Quality assurance is a major issue in the food industry. The authenticity of food ingredients and their traceability are required by consumers and authorities. Plant species such as barley (Hordeum vulgare), rice (Oryza sativa), sunflower (Helianthus annuus), and wheat (Triticum aestivum) are very common among the ingredients of many processed food products; therefore the development of specific assays for their specific detection and quantification are needed. Furthermore, the production and trade of genetically modified lines from an increasing number of plant species brings about the need for control within research, environmental risk assessment, labeling/legal, and consumers' information purposes. We report here the development of four independent real-time polymerase chain reaction (PCR) assays suitable for identification and quantification of four plant species (barley, rice, sunflower, and wheat). These assays target gamma-hordein, gos9, helianthinin, and acetyl-CoA carboxylase sequences, respectively, and were able to specifically detect and quantify DNA from the target plant species. In addition, the simultaneous amplification of RALyase allowed bread from durum wheat to be distinguished. Limits of detection were 1 genome copy for barley, sunflower, and wheat and 3.3 copies for rice real-time PCR systems, whereas limits of quantification were 10 genome copies for barley, sunflower, or wheat and approximately 100 haploid genomes for rice real-time PCR systems. Real-time PCR cycling conditions of the four assays were stated as standard to facilitate their use in routine laboratory analyses. The assays were finally adapted to conventional PCR for detection purposes, with the exception of the wheat assay, which detects rye simultaneously with similar sensitivity in an agarose gel. Publication Types: Research Support, Non-U.S. Gov't PMID: 16131102 [PubMed - indexed for MEDLINE] 910: PLoS Biol. 2005 Sep;3(9):e305. Epub 2005 Aug 30. Candidate gustatory interneurons modulating feeding behavior in the Drosophila brain. Melcher C, Pankratz MJ. Institut für Genetik, Forschungszentrum Karlsruhe, Karlsruhe, Germany. Feeding is a fundamental activity of all animals that can be regulated by internal energy status or external sensory signals. We have characterized a zinc finger transcription factor, klumpfuss (klu), which is required for food intake in Drosophila larvae. Microarray analysis indicates that expression of the neuropeptide gene hugin (hug) in the brain is altered in klu mutants and that hug itself is regulated by food signals. Neuroanatomical analysis demonstrates that hug-expressing neurons project axons to the pharyngeal muscles, to the central neuroendocrine organ, and to the higher brain centers, whereas hug dendrites are innervated by external gustatory receptor-expressing neurons, as well as by internal pharyngeal chemosensory organs. The use of tetanus toxin to block synaptic transmission of hug neurons results in alteration of food intake initiation, which is dependent on previous nutrient condition. Our results provide evidence that hug neurons function within a neural circuit that modulates taste-mediated feeding behavior. Publication Types: Research Support, Non-U.S. Gov't PMID: 16122349 [PubMed - indexed for MEDLINE] 911: Allergy Asthma Proc. 2005 May-Jun;26(3):210-6. Genetically modified and wild soybeans: an immunologic comparison. Yum HY, Lee SY, Lee KE, Sohn MH, Kim KE. Department of Pediatrics, College of Medicine, Pochon CHA University, Pochon, Korea. Most traits introduced into genetically engineered crops result from the expression of new proteins. As the first step toward assessing the allergenic potential of genetically modified organism (GMO) food, immunologic and physicochemical characterizations are needed. We prepared crude extract from GMO soybeans, wild soybeans, curd, and soy milk and then performed sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). After acidification with HCl, the samples were separated to globulin and whey. To evaluate changes in protein composition, either the samples were heated or pepsin was added. Polymerase chain reaction with primer encoding the 35S-promotor and the 3-enol-pyruvyl-shikimat-5-phosphat-synthase gene were performed, respectively, to detect the GMO component. SDS-PAGE results showed definite protein bands at 80 kDa in GMO soybean, 50 kDa in wild soybean, and a similar distribution of protein bands was noticed below 40 kDa. It was difficult to observe protein distribution because of modifications that occurred during processing in soybean-processed products. After heating, proteins of GMO and wild soybeans showed similar distributions and no distinct bands were detected at 50 and 80 kDa. Although SDS-PAGE analyses of raw GMO and wild soybeans differed, the same protein bands of 68, 37, and 20 kDa were observed in the globulin fraction after acidification. After adding pepsin, 20- and 68-kDa bands were found preserved in GMO and wild soybeans. The polymerase chain reaction procedures with primers specific to GMO soybeans showed that GMO soybeans and some curd samples included a GMO component. The skin test results of 49 patients showed 13 positive results to wild soybeans and 8 positive results to GMO soybeans. One patient had a positive skin test result to GMO soybeans only. Sera from nine patients with positive skin tests to the crude extract and a positive capsulated allergen product test to the soybean antigen were used for the immunoblotting of GMO and wild soybeans. GMO soybeans revealed a unique strong immunoglobulin E binding band at 25 kDa in some patients and wild soybeans showed a strong immunoglobulin E binding band at 30-36 kDa. To assess the allergenicity of GMO food, more research, including a selection of controlled sample materials and immunoassays of qualified sera, is needed. Publication Types: Comparative Study Research Support, Non-U.S. Gov't PMID: 16119037 [PubMed - indexed for MEDLINE] 912: Expert Rev Vaccines. 2005 Aug;4(4):591-601. Regulatory issues for plant-made pharmaceuticals and vaccines. Streatfield SJ. Applied Biotechnology Institute, 101 Gateway Boulevard, College Station, TX 77845, USA. sstreatfield@appliedbiotech.org Recombinant plant systems potentially offer economic alternatives to produce large amounts of pharmaceutical proteins, including those used in subunit vaccines. Plant systems also provide a convenient oral delivery option, overcoming the cost and inconvenience of purification and injections. The production of pharmaceutical proteins in transgenic plants is tightly regulated, with the US Department of Agriculture focusing on containment of recombinant material and the US Food and Drug Administration focusing on the production system as it relates to manufacture of the drug or vaccine. Current regulations for the production of plant-made pharmaceuticals are to prevent recombinant proteins from entering the food chain or from persisting in the environment, and to guard against recombinant nucleic acid sequences entering genomes of food or feed crops, or wild species. Several alternative plant production systems are being developed. Each system has its strengths and weaknesses with regard to the economics of production, options for alternative routes of administration, authenticity of products and ease with which the production system can be contained. Risk assessments can be used as a means to quantify risks of inadvertent human or environmental exposure to plant-made pharmaceuticals. Several technologies are being tested that reduce the probability of plant-made pharmaceuticals, or genes encoding them, escaping production sites. Publication Types: Review PMID: 16117714 [PubMed - indexed for MEDLINE] 913: Nat Immunol. 2005 Sep;6(9):857-60. Genetically modified crops and allergenicity. Metcalfe DD. Laboratory of Allergic Diseases, National Institute of Allergy and Infectious Disease, National Institutes of Health, Bethesda, Maryland 20892-1881, USA. dmetcalfe@niaid.nih.gov Publication Types: Review PMID: 16116460 [PubMed - indexed for MEDLINE] 914: Rev Sci Tech. 2005 Apr;24(1):309-22. Current scientific understanding of the environmental biosafety of transgenic fish and shellfish. Kapuscinski AR. Department of Fisheries, Wildlife and Conservation Biology and Institute for Social, Economic and Ecological Sustainability, 200 Hodson Hall, 1980 Folwell Avenue, University of Minnesota, St Paul, MN 55108, USA. A fluorescent zebrafish was the first genetically engineered animal to be marketed, and biotechnologists are developing many transgenic fish and shellfish. Biosafety science is not sufficiently advanced to be able to draw scientifically reliable and broadly trusted conclusions about the environmental effects of these animals. The science is best developed for identifying hazards posed by environmental spread of a transgenic fish or shellfish and least developed for assessing potential ecological harms of spread. Environmental spread of certain transgenic fish or shellfish could be an indirect route of entry into the human food supply. The management of predicted environmental risks is in its infancy and has thus far focused on the first step of the risk management process, i.e. risk reduction, via a few confinement methods. There is a critical need to improve scientific methods of environmental safety assessment and management and to gather empirical data needed to substantiate biosafety conclusions and to effectively manage transgenic fish and shellfish. Scientists and potentially affected parties should participate in prioritising the knowledge gaps to be addressed. Publication Types: Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Review PMID: 16110898 [PubMed - indexed for MEDLINE] 915: Rev Sci Tech. 2005 Apr;24(1):265-74. [Confinement and consumption of cloned and transgenic animals] [Article in French] Houdebine LM, Renard JP. UMR Biologie du Développement et Reproduction, Institut National de la Recherche Agronomique, 78352 Jouyen-Josas, France. Reproduction by cloning can eliminate some of the problems inherent to sexual reproduction, but it creates others. The genetic heritage of nucleus donor cells and the genetic status of clones are not precisely known. Furthermore, reprogramming of the genome of nucleus donor cells by the ovocyte cytoplasm is often incomplete. Animals obtained through cloning are thus essentially genetically identical to their genitors, but they are often epigenetically modified, with unpredictable effects. Transgenesis results in most cases from the addition to a genome of one or more known genes. The direct and indirect effects of transgenesis cannot all be predicted. Specific confinement measures make it possible to raise animals in high-security conditions, preventing their dissemination in the human food chain, in animal feed or in the environment. The toxicity, allergenicity and infectiousness of cloned ortransgenic animals can be evaluated by means of tests. Publication Types: English Abstract Review PMID: 16110894 [PubMed - indexed for MEDLINE] 916: Rev Sci Tech. 2005 Apr;24(1):251-64. Animal cloning: problems and prospects. Wells DN. Reproductive Technologies Group, AgResearch Ruakura, East Street, PB 3123, Hamilton, New Zealand. An efficient animal cloning technology would provide many new opportunities for livestock agriculture, human medicine, and animal conservation. Nuclear cloning involves the production of animals that are genetically identical to the donor cells used in a technique known as nuclear transfer (NT). However, at present it is an inefficient process: in cattle, only around 6% of the embryos transferred to the reproductive tracts of recipient cows result in healthy, longterm surviving clones. Of concern are the high losses throughout gestation, during birth and in the post-natal period through to adulthood. Many of the pregnancy losses relate to failure of the placenta to develop and function correctly. Placental dysfunction may also have an adverse influence on postnatal health. These anomalies are probably due to incorrect epigenetic reprogramming of the donor genome following NT, leading to inappropriate patterns of gene expression during the development of clones. Whilst some physiological tests on surviving clones suggest normality, other reports indicate a variety of post-natal clone-associated abnormalities. This variability in outcome may reflect species-specific and/or cloning methodological differences. Importantly, to date it appears that these clone-associated phenotypes are not transmitted to offspring following sexual reproduction. This indicates that they represent epigenetic errors, rather than genetic errors, which are corrected during gametogenesis. Whilst this needs confirmation at the molecular level, it provides initial confidence in the first application of NT in agriculture, namely, the production of small numbers of cloned sires from genetically elite bulls, for natural mating, to effectively disseminate genetic gain. In addition to the animal welfare concerns with the technology, the underlying health of the animals and the consequential effect on food safety are critical aspects that require investigation to gain regulatory and consumer acceptance. Future improvements in animal cloning will largely arise from a greater understanding of the molecular mechanisms of reprogramming. Publication Types: Review PMID: 16110893 [PubMed - indexed for MEDLINE] 917: Rev Sci Tech. 2005 Apr;24(1):231-42. Traceability of biotech-derived animals: application of DNA technology. Loftus R. IdentiGEN Ltd, Unit 9, Trinity Enterprise Centre, Pearse Street, Dublin 02, Ireland. Traceability is increasingly becoming standard across the agri-food industry, largely driven by recent food crises and the consequent demands for transparency within the food chain. This is leading to the development of a range of traceability concepts and technologies adapted to different industry needs. Experience with genetically modified plants has shown that traceability can play a role in increasing public confidence in biotechnology, and might similarly help allay concerns relating to the development of animal biotechnology. Traceability also forms an essential component of any risk management strategy and is a key requirement for post-marketing surveillance. Given the diversity of traceability concepts and technologies available, consideration needs to be given to the scope and precision of traceability systems for animal biotechnology. Experience to date has shown that conventional tagging and labelling systems can incorporate levels of error and may not have sufficient precision for biotech-derived animals. Deoxyribonucleic acid (DNA) technology can overcome these difficulties by tracing animals and animal by-products through their DNA code rather than an associated label. This offers the possibility of tracing some by-products of animal biotechnology through the supply chain back to source animals, offering unprecedented levels of traceability. Developments in both DNA sampling and analysis technology are making large-scale applications of DNA traceability increasingly cost effective and feasible, and are likely to lead to a broader uptake of DNA traceability concepts. Publication Types: Review PMID: 16110891 [PubMed - indexed for MEDLINE] 918: Rev Sci Tech. 2005 Apr;24(1):201-13. DNA vaccines for aquacultured fish. Lorenzen N, LaPatra SE. Danish Institute for Food and Veterinary Research, Hangovej 2, DK-8200 Aarhus N, Denmark. Deoxyribonucleic acid (DNA) vaccination is based on the administration of the gene encoding the vaccine antigen, rather than the antigen itself. Subsequent expression of the antigen by cells in the vaccinated hosts triggers the host immune system. Among the many experimental DNA vaccines tested in various animal species as well as in humans, the vaccines against rhabdovirus diseases in fish have given some of the most promising results. A single intramuscular (IM) injection of microgram amounts of DNA induces rapid and long-lasting protection in farmed salmonids against economically important viruses such as infectious haematopoietic necrosis virus (IHNV) and viral haemorrhagic septicaemia virus (VHSV). DNA vaccines against other types of fish pathogens, however, have so far had limited success. The most efficient delivery route at present is IM injection, and suitable delivery strategies for mass vaccination of small fish have yet to be developed. In terms of safety, no adverse effects in the vaccinated fish have been observed to date. As DNA vaccination is a relatively new technology, various theoretical and long-term safety issues related to the environment and the consumer remain to be fully addressed, although inherently the risks should not be any greater than with the commercial fish vaccines that are currently used. Present classification systems lack clarity in distinguishing DNA-vaccinated animals from genetically modified organisms (GMOs), which could raise issues in terms of licensing and public acceptance of the technology. The potential benefits of DNA vaccines for farmed fish include improved animal welfare, reduced environmental impacts of aquaculture activities, increased food quality and quantity, and more sustainable production. Testing under commercial production conditions has recently been initiated in Canada and Denmark. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 16110889 [PubMed - indexed for MEDLINE] 919: Rev Sci Tech. 2005 Apr;24(1):149-57. Public perceptions of transgenic animals. Einsiedel EF. Communication Studies Program, University of Calgary, Calgary, Alberta T2N 1N4, Canada. The field of animal biotechnology has been rapidly expanding and the development of transgenic animals has been part of this research expansion. How the public perceives such developments is an important component of policy considerations. In general, biotechnology applications have been judged with evident hierarchies of acceptability. There appearto be hierarchies in terms of the type of organism being modified, the purpose of the application, the means to attain particular ends, and the nature of the benefits obtained. While general awareness of biotechnology and its specific applications remains low to moderate, this article presents data regarding public acceptance of a variety of applications. These range from the use of animals as disease models and as sources for tissues and organs, to the use of transgenic animals for disease control, for food, and for the production of pharmaceutical and industrial products. Case-by-case judgments are evident, but at the same time, the application of criteria such as the nature of the organism being modified, the animal welfare aspects and the ethical-moral concerns are additional criteria for public judgments. These findings are discussed in the context of their implications for public policy. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 16110885 [PubMed - indexed for MEDLINE] 920: Rev Sci Tech. 2005 Apr;24(1):141-8. Risk communication related to animal products derived from biotechnology. McCrea D. Consultant on food and consumer affairs, The Food Consultancy, 127, Havannah Street, Cardiff Bay, Cardiff CF10 5SF, United Kingdom. Previous chapters of this review have dealt with the key considerations related to the application of biotechnology in veterinary science and animal production. This article explores the theory and practice of risk communication and sets out the basic principles for good risk communication when dealing with new technologies, uncertainty, and cautious and sceptical consumers. After failure to communicate with consumers and stakeholders about the risk to human health from bovine spongiform encephalopathy (BSE) in the 1990s, Government Agencies in the United Kingdom have made significant improvements in risk communication. The official inquiry that followed the BSE crisis concluded that a policy of openness was the correct approach, and this article emphasises the importance of consultation, consistency and transparency. There are, however, many different factors that affect public perception of risk (religious, political, social, cultural, etc.) and developing effective risk communication strategies must take all of these complex issues into consideration. Publication Types: Review PMID: 16110884 [PubMed - indexed for MEDLINE] 921: Rev Sci Tech. 2005 Apr;24(1):117-25. Regulatory considerations for biotechnology-derived animals in Canada. Kochhar HP, Adlakha-Hutcheon G, Evans BR. Animal Biotechnology Unit, Animal Products Directorate, Canadian Food Inspection Agency, 59 Camelot Drive, Ottawa, Ontario, K1A 0Y9, Canada Regulatory initiatives relating to biotechnology-derived livestock have focused on animal health, environmental impact, and the general concept of the safety of the food and by-products derived from such animals. Existing regulatory frameworks have been stretched to accommodate these emerging concerns. Public concerns and the expectations of society mean that the regulatory infrastructure is subject to a high level of scrutiny and that regulations are expected to maintain a clear level of confidence, transparency and effectiveness. A sound regulatory regime should be 'neutral', neither 'facilitating' nor 'restricting' the approval of products or by-products derived from biotechnology-derived animals. Publication Types: Review PMID: 16110882 [PubMed - indexed for MEDLINE] 922: Rev Sci Tech. 2005 Apr;24(1):61-74. The safety assessment of foods from transgenic and cloned animals using the comparative approach. Kelly L. Food Standards Australia New Zealand, P.O. Box 7186, Canberra BC, ACT 2610, Australia. The comparative approach to food safety assessment is based on the idea that the safety of a new food can largely be determined by its comparison to the benchmark of commonly consumed foods already in the food supply (also called the 'conventional counterpart'). Any differences between the new food and its conventional counterpart are evaluated to determine their relevance to human health and safety. In this way it is possible to conclude whether a new food is 'as safe as' conventional food already in the food supply. This approach, first developed primarily for use in the safety assessment of food from transgenic plants, is now generally accepted for food from both transgenic and cloned animals as well. This article outlines the basic principles behind the comparative approach, discusses some of the potential food safety concerns associated with transgenic and cloned animals, and describes important elements of the comparative approach and how these might be applied to assessing the safety of food from animals. Publication Types: Comparative Study Review PMID: 16110877 [PubMed - indexed for MEDLINE] 923: Rev Sci Tech. 2005 Apr;24(1):51-60. A framework for the animal health risk analysis of biotechnology-derived animals: a Canadian perspective. Moreau PI, Jordan LT. Animal Health Risk Assessment, Science Branch, Canadian Food Inspection Agency, P.O. Box 11300, Station H, 3851 Fallowfield Road, Ottawa, Ontario K2H 8P9, Canada. This paper describes the framework used by the Canadian Food Inspection Agency to assess the risks to animal health associated with biotechnology-derived animals and their products. In Canada the risks to animal health associated with biotechnology-derived animals are one consideration among several other regulatory concerns (e.g. human health, the environment). The risk analysis process begins with hazard identification, includes a risk assessment for each hazard, and concludes with risk management and risk communication. Publication Types: Review PMID: 16110876 [PubMed - indexed for MEDLINE] 924: J Agric Food Chem. 2005 Aug 24;53(17):6691-6. Identification of genetically modified potato (Solanum tuberosum) cultivars using event specific polymerase chain reaction. Côté MJ, Meldrum AJ, Raymond P, Dollard C. Center for Plant Quarantine Pests, Ottawa Laboratory (Fallowfield), Canadian Food Inspection Agency, 3851 Fallowfield Road, Ottawa, Ontario K2H 8P9, Canada. cotemj@inspection.gc.ca Several genetically modified (GM) cultivars are registered in Canada although they are not currently in commercial production. The GM cultivars can be distinguished from the non-GM and other GM cultivars by analyzing the DNA nucleotide sequence at the insertion site of the transgene corresponding to a single transformation event in the plant genome. Techniques based on modified polymerase chain reaction (PCR) strategies were used to generate sequence information from the plant genome flanking the insertion site of transgenic DNA for specific GM potato events. The plant genome sequence adjacent to the transgenic insertion was used to design PCR primers, which could be used in combination with a primer annealing to one of the nearby inserted genetic elements to amplify an event specific DNA fragment. The event specific PCR fragments generated were sequenced to confirm the specificity of the method. PMID: 16104786 [PubMed - indexed for MEDLINE] 925: Psychol Sci. 2005 Aug;16(8):652-8. The meaning of "natural": process more important than content. Rozin P. Department of Psychology, University of Pennsylvania, Philadephia, 19104, USA. rozin@psych.upenn.edu The meaning of the desirable attribute "natural" was explored in two samples, American college students and adults in the Philadelphia jury pool. Participants rated the naturalness of a variety of "natural" entities, before and after they were transformed by operations such as freezing, adding or removing components, mixing with other natural or unnatural entities, domestication, and genetic engineering. Results support four hypotheses. First, the principle of contagion accounts for many aspects of the reduction of naturalness by contact with unnatural entities. Second, chemical transformations reduce naturalness much more than physical transformations do. Third, the history of an entity's processing is more important in determining its naturalness than is the nature of the entity's contents. Fourth, mixing like natural entities (e.g., water from different sources) does not markedly reduce naturalness. The insertion of a gene from another species, the process used in producing genetically modified organisms, produces the biggest drop in naturalness; domestication, a human-accomplished activity that changes genotype and phenotype in major ways, is considered much less damaging to naturalness. PMID: 16102069 [PubMed - indexed for MEDLINE] 926: Proc Natl Acad Sci U S A. 2005 Aug 30;102(35):12338-43. Epub 2005 Aug 10. Erratum in: Proc Natl Acad Sci U S A. 2005 Dec 13;102(50):18242. Comment in: Proc Natl Acad Sci U S A. 2005 Sep 13;102(37):13003-4. Absence of detectable transgenes in local landraces of maize in Oaxaca, Mexico (2003-2004). Ortiz-García S, Ezcurra E, Schoel B, Acevedo F, Soberón J, Snow AA. Instituto Nacional de Ecología, Secretaría del Medio Ambiente y Recursos Naturales, Colonia Insurgentes Cuicuilco, Delegación Coyoacán, 04530 México D.F., Mexico. In 2000, transgenes were detected in local maize varieties (landraces) in the mountains of Oaxaca, Mexico [Quist, D. & Chapela, I. H. (2001) Nature 414, 541-543]. This region is part of the Mesoamerican center of origin for maize (Zea mays L.), and the genetic diversity that is maintained in open-pollinated landraces is recognized as an important genetic resource of great cultural value. The presence of transgenes in landraces was significant because transgenic maize has never been approved for cultivation in Mexico. Here we provide a systematic survey of the frequency of transgenes in currently grown landraces. We sampled maize seeds from 870 plants in 125 fields and 18 localities in the state of Oaxaca during 2003 and 2004. We then screened 153,746 sampled seeds for the presence of two transgene elements from the 35S promoter of the cauliflower mosaic virus and the nopaline synthase gene (nopaline synthase terminator) from Agrobacterium tumefaciens. One or both of these transgene elements are present in all transgenic commercial varieties of maize. No transgenic sequences were detected with highly sensitive PCR-based markers, appropriate positive and negative controls, and duplicate samples for DNA extraction. We conclude that transgenic maize seeds were absent or extremely rare in the sampled fields. This study provides a much-needed preliminary baseline for understanding the biological, socioeconomic, and ethical implications of the inadvertent dispersal of transgenes from the United States and elsewhere to local landraces of maize in Mexico. Publication Types: Research Support, Non-U.S. Gov't PMID: 16093316 [PubMed - indexed for MEDLINE] 927: Ambio. 2005 Jun;34(4-5):366-70. Consumer preferences for food product quality attributes from Swedish agriculture. Carlsson F, Frykblom P, Lagerkvist CJ. Department of Economics, School of Economics and Commercial Law, Gothenburg University, Sweden. Fredrik.Carlsson@economics.gu.se This paper employs a choice experiment to obtain consumer preferences and willingness to pay for food product quality attributes currently not available in Sweden. Data were obtained from a large mail survey and estimated with a random parameter logit model. We found evidence for intraproduct differences in consumer preferences for identical attributes, as well as interproduct discrepancies in ranking of attributes. Furthermore, we found evidence of a market failure relating to the potential use of genetically modified animal fodder. Finally, we found support for the idea that a cheap-talk script can alleviate problems of external validity of choice experiments. Our results are useful in forming product differentiation strategies within the food industry, as well as for the formation of food policy. Publication Types: Review PMID: 16092270 [PubMed - indexed for MEDLINE] 928: Anal Bioanal Chem. 2005 Sep;383(2):282-90. Epub 2005 Oct 12. Quantitative determination of Roundup Ready soybean (Glycine max) extracted from highly processed flour. Corbisier P, Trapmann S, Gancberg D, Hannes L, Van Iwaarden P, Berben G, Schimmel H, Emons H. Institute for Reference Materials and Measurements, European Commission, Joint Research Centre, Retieseweg 111, 2440, Geel, Belgium. philippe.corbisier@cec.eu.int Roundup Ready soybean powder has been subjected to different amounts of DNA fragmentation to assess the accuracy of real-time PCR on processed food. Certified reference material (CRM) containing 10 g kg(-1) of Roundup Ready soybean (ERM-BF410d) prepared by a dry-mixing processing method was exposed to water at two temperatures, using three different mixing devices, or to baking temperature (250 degrees C) for 30 min. The amount of DNA extracted from the different samples was quantified by fluorimetry. The amount of fragmentation of the extracted DNA was characterised by gel and capillary electrophoresis and the percentage of genetically modified (GM) soybean was determined by a double quantitative real-time PCR method. Measurement of the event GTS 40-3-2 (RUR) was possible in all the treated materials, because small amplicons were amplified. Correct RUR percentages could be measured for intact powders with little or no DNA fragmentation. For samples with a high level of DNA degradation, however, the accuracy of the measurement was found to depend on the method used for DNA extraction. Genomic DNA isolated by use of silica resin resulted in statistically significant overestimation of the amount of GM. PMID: 16091947 [PubMed - indexed for MEDLINE] 929: Wien Klin Wochenschr. 2005 Jul;117(13-14):437-9. [In vitro tests for the determination of allergenic potency of "novel foods' and genetically modified organisms: relevance in vivo?] [Article in German] Jensen-Jarolim E, Untersmayr E. Institut für Physiologie und Pathophysiologie, Medizinische Universität Wien, Wien, Osterreich. erika. jensen-jarolim@meduniwien.ac.at Publication Types: Research Support, Non-U.S. Gov't PMID: 16091867 [PubMed - indexed for MEDLINE] 930: Perspect Biol Med. 2005 Summer;48(3):328-43. Ethical issues in animal cloning. Fiester A. University of Pennsylvania Center for Bioethics, Philadelphia, PA 19104, USA. fiester@mail.med.upenn.edu The issue of human reproductive cloning has recently received a great deal attention in public discourse. Bioethicists, policy makers, and the media have been quick to identify the key ethical issues involved in human reproductive cloning and to argue, almost unanimously, for an international ban on such attempts. Meanwhile, scientists have proceeded with extensive research agendas in the cloning of animals. Despite this research, there has been little public discussion of the ethical issues raised by animal cloning projects. Polling data show that the public is decidedly against the cloning of animals. To understand the public's reaction and fill the void of reasoned debate about the issue, we need to review the possible objections to animal cloning and assess the merits of the anti-animal cloning stance. Some objections to animal cloning (e.g., the impact of cloning on the population of unwanted animals) can be easily addressed, while others (e.g., the health of cloned animals) require more serious attention by the public and policy makers. Publication Types: Review PMID: 16085991 [PubMed - indexed for MEDLINE] 931: Food Chem Toxicol. 2006 Feb;44(2):147-60. Epub 2005 Aug 9. Results of a 90-day safety assurance study with rats fed grain from corn rootworm-protected corn. Hammond B, Lemen J, Dudek R, Ward D, Jiang C, Nemeth M, Burns J. Monsanto Company, 800 North Lindbergh Blvd., St Louis, MO 63167, USA. bruce.g.hammond@monsanto.com The results of a 90-day rat feeding study with YieldGard (YieldGard Rootworm Corn is a registered trademark of Monsanto Technology, LLC.) Rootworm corn (MON 863) grain that is protected against feeding damage caused by corn rootworm larvae are presented. Corn rootworm-protection was accomplished through the introduction of a cry3Bb1 coding sequence into the corn genome for in planta production of a modified Cry3Bb1 protein from Bacillus thuringiensis. Grain from MON 863 and its near isogenic control were separately formulated into rodent diets at levels of 11% and 33% (w/w) by Purina Mills, Inc. Additionally, six groups of rats were fed diets containing grain from different conventional (non-biotechnology-derived) reference varieties. The responses of rats fed diets containing MON 863 were compared to those of rats fed grain from conventional corn varieties. All diets were nutritionally balanced and conformed to Purina Mills, Inc. specifications for Certified LabDiet 5002. There were a total of 400 rats in the study divided into 10 groups of 20 rats/sex/group. Overall health, body weight gain, food consumption, clinical pathology parameters (hematology, blood chemistry, urinalysis), organ weights, gross and microscopic appearance of tissues were comparable between groups fed diets containing MON 863 and conventional corn varieties. This study complements extensive agronomic, compositional and farm animal feeding studies with MON 863 grain, confirming that it is as safe and nutritious as existing conventional corn varieties. PMID: 16084637 [PubMed - indexed for MEDLINE] 932: J Allergy Clin Immunol. 2005 Aug;116(2):403-10. Lack of detectable allergenicity of transgenic maize and soya samples. Batista R, Nunes B, Carmo M, Cardoso C, José HS, de Almeida AB, Manique A, Bento L, Ricardo CP, Oliveira MM. Instituto Nacional de Saúde Dr Ricardo Jorge, Lisboa, Portugal. rbatista@itqb.unl.pt BACKGROUND: The safety issues regarding foods derived from genetically modified (GM) plants are central to their acceptance into the food supply. The potential allergenicity of proteins newly introduced in GM foods is a major safety concern. OBJECTIVE: We sought to monitor, in potentially sensitive human populations, the allergenicity effects of 5 GM materials obtained from sources with no allergenic potential and already under commercialization in the European Union. METHODS: We have performed skin prick tests with protein extracts prepared from transgenic maize (MON810, Bt11, T25, Bt176) and soya (Roundup Ready) samples and from nontransgenic control samples in 2 sensitive groups: children with food and inhalant allergy and individuals with asthma-rhinitis. We have also tested IgE immunoblot reactivity of sera from patients with food allergy to soya (Roundup Ready) and maize (MON810, Bt11, Bt176) samples, as well as to the pure transgenic proteins (CryIA[b] and CP4 5-enolpyruvylshikimate-3-phosphate synthase). RESULTS: None of the individuals undergoing tests reacted differentially to the transgenic and nontransgenic samples under study. None of the volunteers tested presented detectable IgE antibodies against pure transgenic proteins. CONCLUSION: The transgenic products under testing seem to be safe in terms of allergenic potential. We propose postmarket testing as an important screening strategy for putative allergic sensitization to proteins introduced in transgenic plants. Publication Types: Research Support, Non-U.S. Gov't PMID: 16083797 [PubMed - indexed for MEDLINE] 933: Curr Opin Clin Nutr Metab Care. 2005 Sep;8(5):516-22. A perspective on DNA microarray technology in food and nutritional science. Kato H, Saito K, Kimura T. Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, University of Tokyo, Tokyo, Japan. akatoq@mail.ecc.u-tokyo.ac.jp PURPOSE OF REVIEW: The functions of nutrients and other foods have been revealed at the level of gene regulation. The advent of DNA microarray technology has enabled us to analyze the body's response to these factors in a much more holistic manner than before. This review is intended to overview the present status of this DNA microarray technology, hoping to provide food and nutrition scientists, especially those who are planning to introduce this technology, with hints and suggestions. RECENT FINDINGS: The number of papers examining transcriptomics analysis in food and nutrition science has expanded over the last few years. The effects of some dietary conditions and administration of specific nutrients or food factors are studied in various animal models and cultured cells. The target food components range from macronutrients and micronutrients to other functional food factors. Such studies have already yielded fruitful results, which include discovery of novel functions of a food, uncovering hitherto unknown mechanisms of action, and analyses of food safety. SUMMARY: The potency of DNA microarray technology in food and nutrition science is broadly recognized. This technique will surely continue to provide researchers and the public with valuable information on the beneficial and adverse effects of food factors. It should also be acknowledged, however, that there remain problems such as standardization of the data and sharing of the results among researchers in this field. Publication Types: Review PMID: 16079622 [PubMed - indexed for MEDLINE] 934: Environ Health Perspect. 2005 Aug;113(8):A526-33. Comment in: Environ Health Perspect. 2006 Mar;114(3):A146-7. Genetically modified foods: breeding uncertainty. Schmidt CW. Publication Types: News PMID: 16079054 [PubMed - indexed for MEDLINE] 935: Am J Psychiatry. 2005 Aug;162(8):1441-51. Pharmacotherapy and pharmacogenetics of nicotine dependence. Berrettini WH, Lerman CE. Center for Neurobiology and Behavior, Department of Psychiatry, University of Pennsylvania School of Medicine, Clinical Research Bldg., Room 111, 415 Curie Blvd., Philadelphia, PA 19104, USA. wadeb@mail.med.upenn.edu The authors review recent advances in the pharmacotherapy and pharmacogenetics of nicotine dependence. Despite the negative health consequences of smoking, approximately 23% of adults in the United States are daily tobacco smokers and approximately 13% are nicotine dependent. Data for development of new medications for nicotine dependence are likely to come from animal models of the reinforcing value of nicotine, studies to identify proteins in transgenic rodents, and pharmacological studies of nicotine withdrawal. The initial pharmacogenetic studies of pharmacotherapies approved by the United States Food and Drug Administration for treatment of nicotine dependence-nicotine replacement (nicotine gum, nicotine nasal spray, and transdermal nicotine) and bupropion-have identified candidate alleles at the dopamine D2 receptor gene and mu opioid receptor gene that may predict therapeutic response. Because no one medication is likely to be safe and efficacious for a majority of persons with nicotine dependence, it will be useful to develop genetics-based methods and other tools to predict therapeutic response in subgroups of nicotine-dependent persons. Publication Types: Research Support, N.I.H., Extramural Research Support, U.S. Gov't, P.H.S. Review PMID: 16055765 [PubMed - indexed for MEDLINE] 936: Anal Biochem. 2005 Sep 15;344(2):174-82. Development of a peptide nucleic acid polymerase chain reaction clamping assay for semiquantitative evaluation of genetically modified organism content in food. Peano C, Lesignoli F, Gulli M, Corradini R, Samson MC, Marchelli R, Marmiroli N. Department of Environmental Sciences, Division of Genetics and Environmental Biotechnology, University of Parma, 43100, Parma, Italy. In the present study a peptide nucleic acid (PNA)-mediated polymerase chain reaction (PCR) clamping method was developed and applied to the detection of genetically modified organisms (GMO), to test PCR products for band identity and to obtain a semiquantitative evaluation of GMO content. The minimal concentration of PNA necessary to block the PCR was determined by comparing PCRs containing a constant amount of DNA in the presence of increasing concentration of target-specific PNA. The lowest PNA concentration at which specific inhibition took place, by the inhibition of primer extension and/or steric hindrance, was the most efficient condition. Optimization of PCR clamping by PNA was observed by testing five different PNAs with a minimum of 13 bp to a maximum of 15 bp, designed on the target sequence of Roundup Ready soybean. The results obtained on the DNA extracted from Roundup Ready soybean standard flour were verified also on DNA extracted from standard flours of maize GA21, Bt176, Bt11, and MON810. A correlation between the PNA concentration necessary for inducing PCR clamping and the percentage of the GMO target sequence in the sample was found. Publication Types: Evaluation Studies Research Support, Non-U.S. Gov't PMID: 16055074 [PubMed - indexed for MEDLINE] 937: Anal Chem. 2005 Aug 1;77(15):4785-91. High-throughput double quantitative competitive polymerase chain reaction for determination of genetically modified organisms. Mavropoulou AK, Koraki T, Ioannou PC, Christopoulos TK. Laboratory of Analytical Chemistry, Department of Chemistry, University of Athens, Greece 15771. Quantitative competitive polymerase chain reaction (PCR), especially the double competitive PCR methods (DC-PCR), have evolved as reliable approaches to quantification of genetically modified organisms (GMO) in food. However, DC-PCR is a low-throughput method because it requires titration of each sample with various amounts of a competitive internal standard, a protocol that involves several PCRs per sample followed by electrophoresis and densitometry. To address this drawback, we have developed a new method for GMO quantification, namely, a high-throughput double quantitative competitive PCR (HT-DCPCR). In HT-DCPCR, electrophoresis and densitometry are replaced by a rapid, microtiter well-based bioluminometric hybridization assay and there is no need for titration of each sample. The determination of GM soya was chosen as a model. We have constructed internal standards (DNA competitors) both for the 35S promoter sequence and for a plant-specific reference gene (lectin). The competitors have identical size and share the same primer binding sites with the target sequences but differ in a 24-bp internal segment. Each target sequence (35S and lectin) is coamplified with a constant amount (1000 copies) of the respective competitor. The four amplified fragments are hybridized with specific probes and captured on a universal solid phase to achieve simplicity and high throughput. The hybrids are determined by using streptavidin conjugated to the photoprotein aequorin. The ratio of the luminescence values obtained for the target and the competitor is linearly related to the starting amount of target DNA. The limit of quantification for the 35S promoter is 24 copies. The proposed method was evaluated by determining the GMO content of soybean powder certified reference materials. Also HT-DCPCR was compared to real-time PCR in a variety of real samples. Publication Types: Research Support, Non-U.S. Gov't PMID: 16053289 [PubMed - indexed for MEDLINE] 938: Environ Sci Technol. 2005 Jul 1;39(13):280A. GM crop study produces major database. Burke M. PMID: 16053061 [PubMed - indexed for MEDLINE] 939: Science. 2005 Jul 29;309(5735):787-90. An interneuronal chemoreceptor required for olfactory imprinting in C. elegans. Remy JJ, Hobert O. Laboratoire NMDA CNRS UMR 6156, Institut de Biologie du Développement (IBDM), 13288 Marseille Cedex 9, France. remy@ibdm.univ-mrs.fr. Animals alter their behavioral patterns in an experience-dependent manner. Olfactory imprinting is a process in which the exposure of animals to olfactory cues during specific and restricted time windows leaves a permanent memory ("olfactory imprint") that shapes the animal's behavior upon encountering the olfactory cues at later times. We found that Caenorhabditis elegans displays olfactory imprinting behavior that is mediated by a single pair of interneurons. To function in olfactory imprinting, this interneuron pair must express a G protein-coupled chemoreceptor family member encoded by the sra-11 gene. Our study provides insights into the cellular and molecular basis of olfactory imprinting and reveals a function for a chemosensory receptor family member in interneurons. Publication Types: Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. PMID: 16051801 [PubMed - indexed for MEDLINE] 940: Proc Nutr Soc. 2005 Aug;64(3):387-93. The production of very-long-chain PUFA biosynthesis in transgenic plants: towards a sustainable source of fish oils. Napier JA, Sayanova O. Crop Performance and Improvement Division, Rothamsted Research, Harpenden, Herts AL5 2JQ, UK. jonathan.napier@bbsrc.ac.uk There is now considerable evidence of the importance of n-3 long-chain PUFA in human health and development. At the same time, the marine fish stocks that serve as the primary sources of these fatty acids are threatened by continued over-exploitation. Thus, there is an urgent need to provide a sustainable alternative source of the n-3 long-chain PUFA normally found in fish oils. The possibility of using transgenic plants genetically engineered to synthesise these important fatty acids has recently been demonstrated. The approaches taken to realise this outcome will be discussed, as will their prospects for providing a sustainable resource for the future. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 16048673 [PubMed - indexed for MEDLINE] 941: J Environ Sci Health B. 2005;40(4):633-44. Evaluation of detection methods for genetically modified traits in genotypes resistant to European corn borer and herbicides. Ma BL, Subedi K, Evenson L, Stewart G. Eastern Cereal and Oilseed Research Centre, Agriculture and Agri-Food Canada, 960 Carling Ave., Ottawa, ON, Canada K1A 0C6. mab@agr.gc.ca Detection of genetically modified (GM) traits in corn (Zea mays L.) is urgently needed for preservation of genetic identity and marketing GM products. A laboratory study was conducted to evaluate the efficiency, accuracy, and reliability of different analytical methods to detect GM traits in corn. Samples with known fractions of GM concentrations (Bacillus thuringiensis [Bt], Liberty Link [LL] and stacked [Bt/LL] genes) from commercial seed companies and those derived from yellow kernels in a white corn ear, outcrossed by pollen of neighboring Bt hybrid, were tested by lateral flow quick test kits and by enzyme-linked immunosorbent assay (ELISA)-based test strips purchased from different companies. Liberty Link hybrids are resistant to glufosinate (Liberty or Basta) herbicides, whereas Bt corn is developed for the control of European corn borer (Ostrinia nubilalis). Selected samples of GM concentrations were also tested in a commercial laboratory using DNA-based polymerase chain reaction (PCR) test. The results demonstrated that qualitative lateral flow quick tests could detect samples containing 1% or higher Bt and 2% or higher LL concentrations within the recommended time limit of the test. Faint test lines for samples containing 0.5 to 1% Bt or 1 to 2% LL concentrations appeared if samples remained in the test cup overnight. ELISA test strips detected the Bt content semiquantitatively in the range of 0.5 to 2.0%. Grain samples derived from non-Bt corn outcrossed by neighboring Bt pollen had usually lower GM concentrations than commercial GM seed samples. Both ELISA- and DNA-based PCR tests distinguished samples with GM concentrations between 0.1 to 0.5%, but the precision of quantification at this range was very low and results were highly inconsistent. Publication Types: Evaluation Studies Research Support, Non-U.S. Gov't PMID: 16047885 [PubMed - indexed for MEDLINE] 942: J Biotechnol. 2005 Sep 22;119(1):20-5. Isolation of promoter for N-methyltransferase gene associated with caffeine biosynthesis in Coffea canephora. Satyanarayana KV, Kumar V, Chandrashekar A, Ravishankar GA. Plant Cell Biotechnology Department, Central Food Technological Research Institute (CFTRI), Mysore 570020, Karnataka, India. N-Methyltransferases (NMTs) catalyze the three SAM dependent sequential methylation of xanthosine, producing caffeine in Coffea species. In the present work, a PCR based genome walking method was adopted to isolate and clone the promoter for the NMT gene. Inspection of the promoter sequence revealed the presence of several motifs important for the regulation of the gene expression. The whole fragment was fused to the beta-glucuronidase (gus) reporter gene and used in Agrobacterium tumefaciens mediated transformation of Nicotiana tabacum. GUS assays proved that the isolated promoter was able to direct the expression of the reporter gene in transgenic tobacco. Based on the promoter sequence, primer was designed and the genomic fragment comprising the promoter and its corresponding gene was amplified and cloned. Sequencing of one of the genomic clones revealed the presence of four exons and three introns in NMT gene. The differences in the restriction pattern among the genomic clones were studied using PCR-RFLP. This is the first report of cloning of the promoter for a gene involved in caffeine biosynthetic pathway and it opens up the possibility of studying the molecular mechanisms that regulate the production of caffeine. Publication Types: Research Support, Non-U.S. Gov't PMID: 16043251 [PubMed - indexed for MEDLINE] 943: Shokuhin Eiseigaku Zasshi. 2005 Jun;46(3):J203-7. [Consideration of detection method and identification for genetically modified foods] [Article in Japanese] Akiyama H, Matsuda R. National Institute of Health Sciences: 1-18-1, Kamiyoga, Setagaya-ku, Tokyo 158-8501, Japan. Publication Types: Review PMID: 16042304 [PubMed - indexed for MEDLINE] 944: Shokuhin Eiseigaku Zasshi. 2005 Jun;46(3):J193-7. [Standardization of the detection methods for genetically modified organisms in ISO] [Article in Japanese] Futo S. FASMAC Co., Ltd.: 5-1-3, Midorigaoka, Atsugi, Kanagawa 243-0041, Japan. PMID: 16042302 [PubMed - indexed for MEDLINE] 945: Shokuhin Eiseigaku Zasshi. 2005 Jun;46(3):79-85. Detection of genetically modified organisms in foreign-made processed foods containing corn and potato. Monma K, Araki R, Sagi N, Satoh M, Ichikawa H, Satoh K, Tobe T, Kamata K, Hino A, Saito K. Tokyo Metropolitan Institute of Public Health: 3-24-1, Hyakunin-cho, Shinjuku-ku, Tokyo, Japan. Investigations of the validity of labeling regarding genetically modified (GM) products were conducted using polymerase chain reaction (PCR) methods for foreign-made processed foods made from corn and potato purchased in the Tokyo area and in the USA. Several kinds of GM crops were detected in 12 of 32 samples of processed corn samples. More than two GM events for which safety reviews have been completed in Japan were simultaneously detected in 10 samples. GM events MON810 and Bt11 were most frequently detected in the samples by qualitative PCR methods. MON810 was detected in 11 of the 12 samples, and Bt11 was detected in 6 of the 12 samples. In addition, Roundup Ready soy was detected in one of the 12 samples. On the other hand, CBH351, for which the safety assessment was withdrawn in Japan, was not detected in any of the 12 samples. A trial quantitative analysis was performed on six of the GM maize qualitatively positive samples. The estimated amounts of GM maize in these samples ranged from 0.2 to 2.8%, except for one sample, which contained 24.1%. For this sample, the total amount found by event-specific quantitative analysis was 23.8%. Additionally, Roundup Ready soy was detected in one sample of 21 potato-processed foods, although GM potatoes were not detected in any sample. PMID: 16042293 [PubMed - indexed for MEDLINE] 946: Appl Microbiol Biotechnol. 2005 Sep;68(5):588-97. Epub 2005 Oct 26. Biotechnological production and applications of phytases. Haefner S, Knietsch A, Scholten E, Braun J, Lohscheidt M, Zelder O. BASF Aktiengesellschaft, Ludwigshafen, Germany. Phytases decompose phytate, which is the primary storage form of phosphate in plants. More than 10 years ago, the first commercial phytase product became available on the market. It offered to help farmers reduce phosphorus excretion of monogastric animals by replacing inorganic phosphates by microbial phytase in the animal diet. Phytase application can reduce phosphorus excretion by up to 50%, a feat that would contribute significantly toward environmental protection. Furthermore, phytase supplementation leads to improved availability of minerals and trace elements. In addition to its major application in animal nutrition, phytase is also used for processing of human food. Research in this field focuses on better mineral absorption and technical improvement of food processing. All commercial phytase preparations contain microbial enzymes produced by fermentation. A wide variety of phytases were discovered and characterized in the last 10 years. Initial steps to produce phytase in transgenic plants were also undertaken. A crucial role for its commercial success relates to the formulation of the enzyme solution delivered from fermentation. For liquid enzyme products, a long shelf life is achieved by the addition of stabilizing agents. More comfortable for many customers is the use of dry enzyme preparations. Different formulation technologies are used to produce enzyme powders that retain enzyme activity, are stable in application, resistant against high temperatures, dust-free, and easy to handle. Publication Types: Review PMID: 16041577 [PubMed - indexed for MEDLINE] 947: J Insect Physiol. 2005 Oct;51(10):1117-26. Epub 2005 Jul 21. Distribution and residual activity of two insecticidal proteins, avidin and aprotinin, expressed in transgenic tobacco plants, in the bodies and frass of Spodoptera litura larvae following feeding. Christeller JT, Malone LA, Todd JH, Marshall RM, Burgess EP, Philip BA. The Horticulture and Food Research Institute of New Zealand Limited, Palmerston North Research Centre, Private Bag 11030, Palmerston North, New Zealand. jchristeller@hortresearch.co.nz To understand how a major cosmopolitan pest responds to two very different insecticidal proteins and to determine whether herbivorous insects and their frass could be environmental sources of recombinant proteins from transgenic plants, Spodoptera litura (Fab.) (Lepidoptera, Noctuidae) larvae were fed on tobacco leaves expressing either the biotin-binding protein, avidin, or the protease inhibitor, aprotinin. Control larvae received non-transgenic tobacco. Samples of larvae were taken after 5, 6 or 7 days' feeding and frass was collected after two 24-h periods at 6 and 7 days. Insects in all treatments grew significantly during the experiment, but the avidin-fed larvae were significantly smaller than the others on Day 7. Avidin was found in all samples of avidin-fed larvae (7.0+/-0.86 ng mg(-1), n=45), at a lower level than in their frass (31.9+/-5.08 ng mg(-1), n=30), and these frass levels were lower than those of the the leaves fed to the larvae (69.0+/-6.71 ng mg(-1), n=45). All of the avidin detected in these samples was capable of binding biotin. On average, between 10 and 28% of avidin was recovered with the methods used, whereas almost full recovery of aprotinin was effected. Aprotinin levels in larvae (8.2+/-0.53 ng mg(-1), n=45) were also lower than aprotinin levels in frass (77.4+/-6.9 ng mg(-1), n=30), which were somewhat lower than those in the leaves fed to the larvae (88.6+/-2.51 ng mg(-1), n=45). Approximately half the trypsin-binding ability of aprotinin was lost in larvae, and in frass, aprotinin had lost about 90% of its ability to bind trypsin. Publication Types: Research Support, Non-U.S. Gov't PMID: 16039663 [PubMed - indexed for MEDLINE] 948: IUBMB Life. 2005 Apr-May;57(4-5):311-4. Are genetically modified plants useful and safe? Weil JH. Institut de Biologie Moleculaire des Plantes, Strasbourg, France. Jacques-Henry.Weil@ibmp-ulp.u-strasbg.fr So far, plants have been genetically modified essentially to achieve resistance to herbicides, or to pathogens (mainly insects, or viruses), but resistance to abiotic stresses (such as cold, heat, drought, or salt) is also being studied. Genetically modified (GM) plants with improved nutritional qualities have more recently been developed, such as plants containing higher proportions of unsaturated fatty acids (omega-3 and omega-6) in their oil (to prevent cardio-vascular diseases), or containing beta-carotene as in the golden rice (to prevent vitamin A deficiency). Possible risks for human health (such as the production of allergenic proteins), or for the environment (such as the appearance of superweeds as a result from gene flow), should be carefully studied, and a science-based assessment of benefits vs. risks should be made on a case by case basis, both for GM plants and for plants obtained by conventional breeding methods. Publication Types: Review PMID: 16036615 [PubMed - indexed for MEDLINE] 949: Nature. 2005 Jul 21;436(7049):328. Comment on: Nature. 2005 Jun 2;435(7042):561. Unlike climate science, GM is full of uncertainties. Parr D. Publication Types: Comment Letter PMID: 16034396 [PubMed - indexed for MEDLINE] 950: Nature. 2005 Jul 21;436(7049):328. Comment on: Nature. 2005 Jun 2;435(7042):561. Leave GM analysis to the relevant scientists. Couvet D. Publication Types: Comment Letter PMID: 16034395 [PubMed - indexed for MEDLINE] 951: Mol Ecol. 2005 Aug;14(9):2815-23. Uptake of Bt endotoxins by nontarget herbivores and higher order arthropod predators: molecular evidence from a transgenic corn agroecosystem. Harwood JD, Wallin WG, Obrycki JJ. Department of Entomology, University of Kentucky, S-225 Agricultural Science Center North, Lexington, KY 40546-0091, USA. james.harwood@uky.edu The planting of transgenic crops expressing Bacillus thuringiensis endotoxins is widespread throughout the world; the prolific increase in their application exposes nontarget organisms to toxins designed to control pests. To date, studies have focused upon the effects of Bt endotoxins on specific herbivores and detritivores, without consideration of their persistence within arthropod food webs. Here, we report the first quantitative field evaluation of levels of Bt endotoxin within nontarget herbivores and the uptake by higher order arthropods. Antibody-based assays indicated significant quantities of detectable Cry1Ab endotoxin within nontarget herbivores which feed on transgenic corn (including the corn flea beetle, Chaetocnema pulicaria, Japanese beetle, Popillia japonica and southern corn rootworm, Diabrotica undecimpunctata howardi). Furthermore, arthropod predators (Coccinellidae, Araneae, and Nabidae) collected from these agroecosystems also contained significant quantities of Cry1Ab endotoxin indicating its movement into higher trophic levels. This uptake by predators is likely to have occurred by direct feeding on plant material (in predators which are facultatively phytophagous) or the consumption of arthropod prey which contained these proteins. These data indicate that long-term exposure to insecticidal toxins occurs in the field. These levels of exposure should therefore be considered during future risk assessments of transgenic crops to nontarget herbivores and arthropod predators. Publication Types: Comparative Study Research Support, Non-U.S. Gov't PMID: 16029480 [PubMed - indexed for MEDLINE] 952: Sci Prog. 2004;87(Pt 4):227-47. The determinants of lifespan in the nematode Caenorhabditis elegans: a short primer. Geanacopoulos M. Mark Geanacopoulos is based at the National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, 2 Democracy Plaza, Rm 642B, 6707 Democracy Boulevard, MSC 5458, Bethesda, MD 20892-5458, USA. GeanacoM@intra.niddk.nih.gov Transparent, easily-maintained, amenable to genetic manipulation, and living for only a few weeks, the nematode Caenorhabditis elegans is a leading animal model for the study of the determinants of lifespan. The original genetic screen for increased longevity identified a mutant, age-1, with a defect in one component of a signal transduction pathway. This pathway functioned as a genetic switch and governed the decision whether to enter a specialized larval form, dauer, that enables the worm to withstand the scarcity of food or other stressful conditions. These age-1 worms had an increased tendency to become dauers, but if they did not adopt the dauer developmental pathway, they lived longer than wild type worms. age-1 and other longevity mutants with dauer phenotypes are vigorous, indicating that they do not suffer from a significant energy deficit, and stress resistant. Mutation of genes encoding mitochondrial components was found to be another means of extending the lifespan of the worm, although the associated phenotypes suggest a deficiency of available energy. While there are now many documented genetic manipulations which can extend the worm's lifespan, it has been difficult to come to definite conclusions as to the mechanism(s) by which lifespan is extended. The most carefully studied mutant strains have complex changes in gene expression and metabolism making it difficult to ascertain what changes are critical. The free radical theory of aging is the dominant biochemical theory of aging, and the phenotypes of the well-characterized longevity mutants worm can be accommodated to it. However discrete interventions to lower reactive oxygen species, or mitigate their effects, have not produced consistent easily-interpretable results in terms of lifespan extension. It has become clear that the insulin-dependent signalling mechanism that regulates lifespan in the worm functions in the context of a complex endocrine system and the hormonal control of aging is an emerging focus of research in worms and higher organisms. Publication Types: Review PMID: 16028834 [PubMed - indexed for MEDLINE] 953: Environ Biosafety Res. 2004 Oct-Dec;3(4):225-31. Spider web survey or whole plant visual sampling? Impact assessment of Bt corn on non-target predatory insects with two concurrent methods. Tóth F, Arpás K, Szekeres D, Kádár F, Szentkirályi F, Szénási A, Kiss J. Szent István University, Faculty of Agricultural and Environmental Sciences, Department of Plant Protection, H-2103 Gödöllö, Páter K. u. 1, Hungary. toth.ferenc@mkk.szie.hu Impact assessment studies rely on relevant sampling methods. Correct evaluation of methods can be done by their concurrent use in the same sampling site and period. Collecting webs of Theridion impressum L. Koch (Araneae: Theridiidae) may serve as an arthropod sampling method: empty cuticles of prey items remain attached to the back side of the leaves with adhesive silk. Our study aimed to compare the applicability of the two methods concurrently (spider web survey and whole plant visual sampling) in a risk assessment study. The spider web survey recorded more predatory insect families than the whole plant visual sampling. Both methods were able to detect significant differences in the quantity of predatory insects in Bt vs. isogenic plots, but not in the same taxa (Nabidae: spider-web, 2001, Bt > Iso; Coccinellidae: plant sampling, 2001, Iso > Bt; Welsh-test, P < 0.05); thus, they could not confirm each other. The lack of confirmation can be explained by differences in the sensitivity and selectivity of the two methods. A web survey of T. impressum has the practical advantage that although we concentrate only on the one species during field sampling, we gain additional information on a wide range of foliage-dwelling arthropods. Due to several biological uncertainties, interpretation and explanation of the results remain problematic. Thus, additional research--based on in-situ observation and recording of T. impressum-prey interactions--is necessary before we could propose web survey method as a complementary tool in ecological impact assessment. Publication Types: Research Support, Non-U.S. Gov't PMID: 16028799 [PubMed - indexed for MEDLINE] 954: Environ Biosafety Res. 2004 Oct-Dec;3(4):215-23. Effect of food components and processing parameters on DNA degradation in food. Bauer T, Hammes WP, Haase NU, Hertel C. Institute of Food Technology, University of Hohenheim, 70593 Stuttgart, Germany. The effect of food components on degradation of DNA by DNase I (EC 3.1.21.1) was monitored by electrotransformation of Escherichia coil, making it possible to determine the number of plasmid molecules capable of giving rise to transformed cells. The transformation frequency increased linearly with the plasmid number within the range of 2 x 10(6) to 2 x 10(10). DNA degradation was reduced by one order of magnitude in the presence of 0.05% (w.v(-1)) maltol or 1 mM putrescine. Complete inhibition of degradation was observed with > or = 0.2% (w.v(-1)) maltol, > or = 0.01% (w.v(-1)) octyl gallate or > or = 0.5 mM of spermine. To monitor degradation of plant DNA during food processing, a real-time PCR system was established. The ratio of copy numbers of a potato gbss DNA fragment of 325 bp and a nested 96 bp fragment was determined. The latter served as internal reference for normalization. The system made it possible to exclude process-dependent changes of DNA concentration in the food matrix. Processing of genetically modified potatoes to dried potato sticks, crisps or flakes was studied and drying steps were shown to exert the strongest effect on DNA degradation, resulting in a drop of the ratio from 0.73 to 0.16. Publication Types: Research Support, Non-U.S. Gov't PMID: 16028798 [PubMed - indexed for MEDLINE] 955: Nutr Rev. 2005 Jun;63(6 Pt 1):210-23. Science, law, and politics in FDA's genetically engineered foods policy: scientific concerns and uncertainties. Pelletier DL. Division of Nutritional Sciences, Cornell University, 378 MVR Hall, Ithaca, NY 14853, USA. dlp5@cornell.edu The Food and Drug Administration's (FDA's) 1992 policy statement granted genetically engineered foods presumptive GRAS (generally recognized as safe) status. Since then, divergent views have been expressed concerning the scientific support for this policy. This paper examines four sources to better understand the basis for these claims: 1) internal FDA correspondence; 2) reports from the National Academy of Sciences; 3) research funded by US Department of Agriculture from 1981 to 2002; and 4) FDA's proposed rules issued in 2001. These sources reveal that little research has been conducted on unintended compositional changes from genetic engineering. Profiling techniques now make this feasible, but the new debate centers on the functional meaning of compositional changes. Publication Types: Review PMID: 16028565 [PubMed - indexed for MEDLINE] 956: J Trace Elem Med Biol. 2005;18(4):333-8. Iron content and bioavailability in rice. Meng F, Wei Y, Yang X. Ministry of Education Key Laboratory of Environmental Remediation and Ecosystem Health, Zhejiang University, Huajiachi Campus, Hangzhou 310029, China. Iron deficiency is probably the most widespread micronutrient deficiency in humans. Since rice is the main staple food for more than half of the global population, improving the iron content and bioavailability in rice is a perspective and an effective way to alleviate or even solve this problem. The present paper evaluates the iron content in different cereal foods (black rice, rice, red rice, sticky rice and millet) and different rice seeds as well as in the milling products, and the iron bioavailability of different forms. The data show that the iron content in black rice is higher than in the other rice types, and in rice chaff and husk the content is still fairly high. However, the iron content in rice and fine rice, which are the people's main staple food, is fairly low. As to the bioavailability of iron, it is fairly low in vegetable foods, almost at the level of 10%. Several methods have been applied to improve iron content and bioavailability in rice seed. Apart from breeding and genetic engineering, biochemical and physical approaches have frequently been used as prospective methods to regulate iron content and bioavailability in rice grains. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 16028495 [PubMed - indexed for MEDLINE] 957: J Trace Elem Med Biol. 2005;18(4):299-307. Agriculture: the real nexus for enhancing bioavailable micronutrients in food crops. Welch RM, Graham RD. USDA-ARS, US Plant, Soil and Nutrition Laboratory, Tower Road, Cornell University, Ithaca, NY 14853-2901, USA. rmw1@cornell.edu Human existence requires that agriculture provide at least 50 nutrients (e.g., vitamins, minerals, trace elements, amino acids, essential fatty acids) in amounts needed to meet metabolic demands during all seasons. If national food systems do not meet these demands, mortality and morbidity rates increase, worker productivity declines, livelihoods are diminished and societies suffer. Today, many food systems within the developing world cannot meet the nutritional needs of the societies they support mostly due to farming systems that cannot produce enough micronutrients to meet human needs throughout the year. Nutrition transitions are also occurring in many rapidly developing countries that are causing chronic disease (e.g., cancer, heart disease, stroke, diabetes, and osteoporosis) rates to increase substantially. These global developments point to the need to explicitly link agricultural technologies to human health. This paper reviews some ways in which agriculture can contribute significantly to reducing micronutrient malnutrition globally. It concludes that it is imperative that close linkages be forged between the agriculture, nutrition and health arenas in order to find sustainable solutions to micronutrient malnutrition with agriculture becoming the primary intervention tool to use in this fight. Publication Types: Review PMID: 16028491 [PubMed - indexed for MEDLINE] 958: Genet Med. 2005 Jul-Aug;7(6):454-5. Genetic testing, biotechnology, and GMOs: A snapshot of public opinion, 2003 through 2004. Alford RL, Morris KE, Rives CM, Scherer SE, Weinstock G, Gibbs RA, Ghonima K, Belcher M, Valdes H, Sumners C, Law C, Reiff P. Publication Types: Letter Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. PMID: 16024979 [PubMed - indexed for MEDLINE] 959: Proc Biol Sci. 2005 May 22;272(1567):1031-8. The cost of resistance to Bacillus thuringiensis varies with the host plant of Trichoplusia ni. Janmaat AF, Myers JH. Department of Biological Sciences, 8888 University Drive, Simon Fraser University, Burnaby, BC, V5A 1S6, Canada. alida_janmaat@sfu.ca Selection for resistance to insecticides, diseases and parasitoids is assumed to be costly and often requires tradeoffs with reproductive fitness. The costs of resistance, however, are often difficult to measure. Cabbage looper, Trichoplusia ni, a generalist Lepidopteran herbivore, has become highly resistant following the extensive use of the microbial insecticide, Bacillus thuringiensis kurstaki (Bt) in vegetable greenhouses. We compared the growth rate, pupal size and survival of resistant, susceptible and hybrid T. ni larvae fed on tomato, bell pepper and cucumber. Performance was best on cucumber and worst on pepper, and the magnitude of fitness costs associated with Bt resistance increased with declining host plant suitability. This supports the hypothesis that in this system, resistance costs are condition dependent and are greatest in the most stressful environment. Management strategies that rely on the presence of fitness costs to reduce the frequency of resistance genes must consider this variation and should be more successful on crops that are less suitable food plants. In general, condition dependence should be considered in studies designed to measure the costs of resistance. Publication Types: Comparative Study Research Support, Non-U.S. Gov't PMID: 16024361 [PubMed - indexed for MEDLINE] 960: Lancet. 2005 Jul 16-22;366(9481):202-3. Comment on: Lancet. 2005 May 21-27;365(9473):1746. Ending hunger in China. Goklany IM. Publication Types: Comment Letter PMID: 16023506 [PubMed - indexed for MEDLINE] 961: Transgenic Res. 2005 Apr;14(2):167-78. Production of biologically active GM-CSF in sugarcane: a secure biofactory. Wang ML, Goldstein C, Su W, Moore PH, Albert HH. Hawaii Agriculture Research Center, Aiea, HI 96701, USA. Over 300 transgenic sugarcane plants representing approx. 200 independent lines producing the human cytokine granulocyte macrophage colony stimulating factor (GM-CSF) were analyzed for recombinant protein accumulation and activity levels. Expression constructs differed in use of the maize polyubiquitin 1, Mubi-1, or the sugarcane polyubiquitin 9, SCubi9, promoters; presence or absence of a C-terminal HDEL tag for ER retention; and presence or absence of a 6X Histidine tag for metal ion affinity purification. Accumulation of GM-CSF protein ranged from undetectable to 0.02% of total soluble protein. No significant difference was observed between the two promoters; however, the ER retention tag was required for higher accumulation levels. Human bone marrow cells (TF-1), which require GM-CSF for cell division, proliferated when growth media was supplemented with transgenic sugarcane extracts. Comparison to purified commercially produced GM-CSF indicated the sugarcane-produced protein had essentially identical activity levels. In a 14-month field trial, accumulation levels remained stable. This is the first report of field production of GM-CSF. During the field trial, no flowering of the trial plants occurred; no pollen or seed was produced. Drying, burning, and burial of the test plants effectively blocked possible routes for the transgenic sugarcane to enter the environment or food supply. Sugarcane may provide a highly secure system for biofactory production of pharmaceutical proteins. Publication Types: Comparative Study Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. PMID: 16022388 [PubMed - indexed for MEDLINE] 962: Transgenic Res. 2005 Apr;14(2):159-65. Enhancement of EPA and DHA biosynthesis by over-expression of masu salmon delta6-desaturase-like gene in zebrafish. Alimuddin, Yoshizaki G, Kiron V, Satoh S, Takeuchi T. Department of Marine Biosciences, Tokyo University of Marine Science and Technology, Minato, Tokyo 108-8477, Japan. goro@s.kaiyodai.ac.jp The n - 3 polyunsaturated fatty acids, especially eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) have important nutritional benefits in humans. Farmed fish could serve as promising sources of EPA/DHA, but they need these fatty acids or their precursors in their diets. Here we transferred masu salmon delta6-desaturase-like gene in zebrafish to increase its ability for synthesizing EPA and DHA. Expression of this gene in transgenic fish elevated their EPA content by 1.4-fold and DHA by 2.1-fold. On the other hand, the alpha-linolenic acid (ALA) content decreased, it being a substrate of delta6-desaturase, while the total lipid remained constant. This achievement demonstrates that fatty acid metabolic pathway in fish can be modified by the transgenic technique, and perhaps this could be applied to tailor farmed fish as even better sources of valuable human food. Publication Types: Research Support, Non-U.S. Gov't PMID: 16022387 [PubMed - indexed for MEDLINE] 963: Transgenic Res. 2005 Apr;14(2):133-44. Effects of Bt maize on the herbivore Spodoptera littoralis (Lepidoptera: Noctuidae) and the parasitoid Cotesia marginiventris (Hymenoptera: Braconidae). Vojtech E, Meissle M, Poppy GM. School of Biological Sciences, University of Southampton, Basset Crescent East, Southampton SO16 7PX, UK. Recent studies have shown that transgenic insect resistant plants can have negative effects on non-target herbivores as well as on beneficial insects. The study of tritrophic interactions gives insight into the complex mechanisms of food webs in the field and can easily be incorporated into a tiered risk assessment framework. We investigated the effects of transgenic maize (Zea mays) expressing insecticidal proteins derived from Bacillus thuringiensis (Bt maize) on Spodoptera littoralis, a non-target herbivore, and on the hymenopteran parasitoid Cotesia marginiventris. In a laboratory study, S. littoralis larvae were reared for their whole lifespan on a mixture of leaves and stems from 2-4-week old Bt maize plants. S. littoralis survival, developmental times and larval weights were significantly affected by Bt maize diet. However, adult moths, which survived development on Bt maize, were the same size as the adults from the control group. C. marginiventris survival, developmental times and cocoon weights were significantly negatively affected if their S. littoralis host larva had been fed Bt maize. ELISA tests confirmed that S. littoralis larvae ingest high amounts of CrylA(b) toxin while feeding on Bt maize. In S. littoralis pupae and in C. marginiventris cocoon silk, only traces of the toxin could be detected. No toxin was found in S. littoralis and C. marginiventris adults. Thus the toxin is not accumulating in the trophic levels and in fact appears to be excreted. Our results suggest that the effects on C. marginiventris when developing in susceptible S. littoralis larvae are indirect (host mediated). The biological relevance of those results and the significance of this study in risk assessment are discussed. Publication Types: Research Support, Non-U.S. Gov't PMID: 16022385 [PubMed - indexed for MEDLINE] 964: Appetite. 2005 Aug;45(1):40-6. Food and the relation between values and attitude characteristics. Dreezens E, Martijn C, Tenbült P, Kok G, de Vries NK. Department of Experimental Psychology, Maastricht University, P.O. Box 616, 6200 MD Maastricht, The Netherlands. e.dreezens@psychology.unimaas.nl This survey showed that the values power (dominance over nature and resources) and universalism (respect for people and for nature) are related to attitudes toward genetically modified food (GMF) and organically grown food (OGF). Furthermore, these values have an influence on the centrality, commitment and ambivalence of these attitudes. Values that are positively related to an attitude influence how central this attitude is to a person. However, values that are negatively related to an attitude have a larger effect on the commitment of this attitude. No such pattern of effects was found for the relationship between ambivalence and values. These data suggest that centrality, commitment, and ambivalence are structurally different constructs that have a distinct relationship with specific values. Publication Types: Research Support, Non-U.S. Gov't PMID: 16018906 [PubMed - indexed for MEDLINE] 965: Shokuhin Eiseigaku Zasshi. 2005 Apr;46(2):55-7. [Study on positive control for GM papaya (55-1) detection method by GUS (beta-glucuronidase) assay] [Article in Japanese] Takahashi K, Horie M. Saitama Institute of Public Health: 639-1 Kamiokubo, Sakura-ku, Saitama-shi, Saitama 338-0824, Japan. A suitable positive control was investigated for histochemical assay (GUS-examining method) to detect genetically modified (GM) papaya (55-1), currently undergoing a safety assessment in Japan. Six different kinds of test papers were soaked with beta-glucuronidase solution and examined for GUS activity. The test papers made of nylon and glass fiber turned blue, and were stable for fifteen months at -20 degrees C. They are concluded to be useful as positive controls in the GUS-examining method for inspection of GM papaya (55-1). Publication Types: English Abstract PMID: 16018592 [PubMed - indexed for MEDLINE] 966: America (NY). 2005 May 2;192(14):8-10. Genetic engineering is not the answer. McDonagh S. PMID: 16018095 [PubMed - indexed for MEDLINE] 967: Rev Med Chir Soc Med Nat Iasi. 2004 Oct-Dec;108(4):838-42. [Genetically modified foods. Advantages and human health risks] [Article in Romanian] Filip L, Miere D, Indrei LL. Universitatea de Medicină şi Farmacie Iuliu Haţieganu Cluj-Napoca Facultatea de Farmacie, Disciplina de Chimie sanitară. One of the most important issue with which the mankind is confronting now is related to the quantitatively as well as qualitatively assurance of the food supply necessary for human species existence. In this context, by means of genetic engineering, modified genetic organisms were obtained. In the first stage, plant crops with high productivity and resistant against diseases and pests were obtained. After that, food products having modified organoleptic properties and high nutrition values were produced. The main problem concerning the long-term consumption of these products is their toxicity, which until now was not confirmed or denied. For this reason, tests are necessary to be made in order to stipulate and prevent these effects. Publication Types: English Abstract Review PMID: 16004228 [PubMed - indexed for MEDLINE] 968: Nat Biotechnol. 2005 Jul;23(7):785; author reply 787-9. Comment on: Nat Biotechnol. 2005 Apr;23(4):439-44. Regulatory regimes for transgenic crops. Wilson A, Latham J, Steinbrecher R. Publication Types: Comment Letter PMID: 16003352 [PubMed - indexed for MEDLINE] 969: Nat Biotechnol. 2005 Jul;23(7):785-7; author reply 787-9. Comment on: Nat Biotechnol. 2005 Apr;23(4):439-44. Regulatory regimes for transgenic crops. Schubert D. Publication Types: Comment Letter PMID: 16003351 [PubMed - indexed for MEDLINE] 970: J AOAC Int. 2005 May-Jun;88(3):877-87. Chromatographic determination of amino acids in foods. Peace RW, Gilani GS. Health Canada, Nutrition Research Division, Food Directorate, Health Products and Food Branch, Ottawa, ON, K1A 0L2, Canada. bob_peace@hc-sc.gc.ca Amino acids in foods exist in a free form or bound in peptides, proteins, or nonpeptide bonded polymers. Naturally occurring L-amino acids are required for protein synthesis and are precursors for essential molecules, such as co-enzymes and nucleic acids. Nonprotein amino acids may also occur in animal tissues as metabolic intermediates or have other important functions. The development of bacterially derived food proteins, genetically modified foods, and new methods of food processing; the production of amino acids for food fortification; and the introduction of new plant food sources have meant that protein amino acids and amino acid enantiomers in foods can have both nutritional and safety implications for humans. There is, therefore, a need for the rapid and accurate determination of amino acids in foods. Determination of the total amino acid content of foods requires protein hydrolysis by various means that must take into account variations in stability of individual amino acids and resistance of different peptide bonds to the hydrolysis procedures. Modern methods for separation and quantitation of free amino acids either before or after protein hydrolysis include ion exchange chromatography, high performance liquid chromatography (LC), gas chromatography, and capillary electrophoresis. Chemical derivatization of amino acids may be required to change them into forms amenable to separation by the various chromatographic methods or to create derivatives with properties, such as fluorescence, that improve their detection. Official methods for hydrolysis and analysis of amino acids in foods for nutritional purposes have been established. LC is currently the most widely used analytical technique, although there is a need for collaborative testing of methods available. Newer developments in chromatographic methodology and detector technology have reduced sample and reagent requirements and improved identification, resolution, and sensitivity of amino acid analyses of food samples. Publication Types: Review PMID: 16001866 [PubMed - indexed for MEDLINE] 971: J AOAC Int. 2005 May-Jun;88(3):814-22. Real-time polymerase chain reaction detection of cauliflower mosaic virus to complement the 35S screening assay for genetically modified organisms. Cankar K, Ravnikar M, Zel J, Gruden K, Toplak N. National Institute of Biology, Department of Plant Physiology and Biotechnology, Vecna pot 111, 1000 Ljubljana, Slovenia. katja.cankar@nib.si Labeling of genetically modified organisms (GMOs) is now in place in many countries, including the European Union, in order to guarantee the consumer's choice between GM and non-GM products. Screening of samples is performed by polymerase chain reaction (PCR) amplification of regulatory sequences frequently introduced into genetically modified plants. Primers for the 35S promoter from Cauliflower mosaic virus (CaMV) are those most frequently used. In virus-infected plants or in samples contaminated with plant material carrying the virus, false-positive results can consequently occur. A system for real-time PCR using a TaqMan minor groove binder probe was designed that allows recognition of virus coat protein in the sample, thus allowing differentiation between transgenic and virus-infected samples. We measured the efficiency of PCR amplification, limits of detection and quantification, range of linearity, and repeatability of the assay in order to assess the applicability of the assay for routine analysis. The specificity of the detection system was tested on various virus isolates and plant species. All 8 CaMV isolates were successfully amplified using the designed system. No cross-reactivity was detected with DNA from 3 isolates of the closely related Carnation etched ring virus. Primers do not amplify plant DNA from available genetically modified maize and soybean lines or from different species of Brassicaceae or Solanaceae that are natural hosts for CaMV. We evaluated the assay for different food matrixes by spiking CaMV DNA into DNA from food samples and have successfully amplified CaMV from all samples. The assay was tested on rapeseed samples from routine GMO testing that were positive in the 35S screening assay, and the presence of the virus was confirmed. Publication Types: Research Support, Non-U.S. Gov't PMID: 16001857 [PubMed - indexed for MEDLINE] 972: Plant J. 2005 Jul;43(2):299-308. Virus-induced gene silencing in tomato fruit. Fu DQ, Zhu BZ, Zhu HL, Jiang WB, Luo YB. Laboratory of Fruit Biology, College of Food Science & Nutritional Engineering, China Agricultural University, No. 17 Qinghua East Road, Beijing 100083, China. Virus-induced gene silencing (VIGS) is a powerful tool for the study of gene function in plants. Here we report that either by syringe-infiltrating the tobacco rattle virus (TRV)-vector into the surface, stem or carpopodium of a tomato fruit attached to the plant or by vacuum-infiltrating into a tomato fruit detached from the plant, TRV can efficiently spread and replicate in the tomato fruit. Although VIGS can be performed in tomato fruit by all of the means mentioned above, the most effective method is to inject the TRV-vector into the carpopodium of young fruit attached to the plant about 10 days after pollination. Several reporter genes related to ethylene responses and fruit ripening, including LeCTR1 and LeEILs genes, were also successfully silenced by this method during fruit development. In addition, we found that the silencing of the LeEIN2 gene results in the suppression of tomato fruit ripening. The results of our study indicate that the application of VIGS techniques by the described methods can be successfully applied to tomato fruit and is a valuable tool for studying functions of the relevant genes during fruit developing. Publication Types: Research Support, Non-U.S. Gov't PMID: 15998315 [PubMed - indexed for MEDLINE] 973: J Ind Microbiol Biotechnol. 2005 Dec;32(11-12):502-13. Epub 2005 Jul 2. Strategies for the engineered phytoremediation of toxic element pollution: mercury and arsenic. Meagher RB, Heaton AC. Department of Genetics, University of Georgia, Athens, GA 30602, USA. meagher@uga.edu Plants have many natural properties that make them ideally suited to clean up polluted soil, water, and air, in a process called phytoremediation. We are in the early stages of testing genetic engineering-based phytoremediation strategies for elemental pollutants like mercury and arsenic using the model plant Arabidopsis. The long-term goal is to develop and test vigorous, field-adapted plant species that can prevent elemental pollutants from entering the food-chain by extracting them to aboveground tissues, where they can be managed. To achieve this goal for arsenic and mercury, and pave the way for the remediation of other challenging elemental pollutants like lead or radionucleides, research and development on native hyperaccumulators and engineered model plants needs to proceed in at least eight focus areas: (1) Plant tolerance to toxic elementals is essential if plant roots are to penetrate and extract pollutants efficiently from heterogeneous contaminated soils. Only the roots of mercury- and arsenic-tolerant plants efficiently contact substrates heavily contaminated with these elements. (2) Plants alter their rhizosphere by secreting various enzymes and small molecules, and by adjusting pH in order to enhance extraction of both essential nutrients and toxic elements. Acidification favors greater mobility and uptake of mercury and arsenic. (3) Short distance transport systems for nutrients in roots and root hairs requires numerous endogenous transporters. It is likely that root plasma membrane transporters for iron, copper, zinc, and phosphate take up ionic mercuric ions and arsenate. (4) The electrochemical state and chemical speciation of elemental pollutants can enhance their mobility from roots up to shoots. Initial data suggest that elemental and ionic mercury and the oxyanion arsenate will be the most mobile species of these two toxic elements. (5) The long-distance transport of nutrients requires efficient xylem loading in roots, movement through the xylem up to leaves, and efficient xylem unloading aboveground. These systems can be enhanced for the movement of arsenic and mercury. (6) Aboveground control over the electrochemical state and chemical speciation of elemental pollutants will maximize their storage in leaves, stems, and vascular tissues. Our research suggests ionic Hg(II) and arsenite will be the best chemical species to trap aboveground. (7) Chemical sinks can increase the storage capacity for essential nutrients like iron, zinc, copper, sulfate, and phosphate. Organic acids and thiol-rich chelators are among the important chemical sinks that could trap maximal levels of mercury and arsenic aboveground. (8) Physical sinks such as subcellular vacuoles, epidermal trichome cells, and dead vascular elements have shown the evolutionary capacity to store large quantities of a few toxic pollutants aboveground in various native hyperaccumulators. Specific plant transporters may already recognize gluthione conjugates of Hg(II) or arsenite and pump them into vacuole. Publication Types: Research Support, U.S. Gov't, Non-P.H.S. Review PMID: 15995854 [PubMed - indexed for MEDLINE] 974: BMJ. 2005 Jul 2;331(7507):11. Developing countries may not get benefits of GM food. Brettingham M. Publication Types: News PMID: 15994680 [PubMed - indexed for MEDLINE] 975: J Environ Health. 2005 Jun;67(10):44-9. Health professionals hold positive attitudes toward biotechnology and genetically engineered foods. Schmidt J, Vickery CE, Cotugna NA, Snider OS. Maryland Agricultural Education Foundation, Sudlersville, MD 21668, USA. jenhans@dmv.com Few biotechnology processes have elicited the degree of controversy that genetic manipulation of food through recombinant DNA technology has. Research has shown that consumers turn to health professionals for answers to questions regarding health and nutrition. This study sought to assess the knowledge, attitudes, and beliefs of physicians (MDs/DOs), nurse practitioners (NPs), and registered dietitians (RDs) toward food biotechnology and genetic engineering (GE). Six hundred three-part, self-administered surveys were sent to health professionals holding active professional licenses. Statistical analysis included analysis of variance with Tukey's HSD and Scheffe's post hoc tests. Attitudes toward GE were positive. MDs held more positive attitudes than NPs or RDs (p = .000). MDs and NPs supported the use of GE to improve plant resistance to pests; RDs tended to support nutritional-improvement technology. All groups supported the use of GE to produce human medicines and the current Food and Drug Administration (FDA) labeling policy. No profession was more knowledgeable than another. Biotechnology holds the potential to positively affect human health. All health professionals can facilitate or diminish this process through their understanding of the technology and their ability to communicate effectively about the science and issues associated with biotechnology. Publication Types: Comparative Study Research Support, Non-U.S. Gov't PMID: 15991684 [PubMed - indexed for MEDLINE] 976: Plant Mol Biol. 2005 Mar;57(5):645-57. Suppression of the cysteine protease, aleurain, delays floret senescence in Brassica oleracea. Eason JR, Ryan DJ, Watson LM, Hedderley D, Christey MC, Braun RH, Coupe SA. NZ Institute for Crop and Food Research, Private Bag 11 600, Palmerston North, New Zealand. easonj@crop.cri.nz An aleurain-like protein, BoCP5, is up-regulated during harvest-induced senescence in broccoli floret and leaf tissue. BoCP5 is most closely related to an Arabidopsis protein (91%, AAF43041) and has 71% identity to barley aleurain (P05167). The mRNA for this gene accumulates within 6 h after harvest in broccoli florets, and its expression is reduced in tissue that has been held in senescence-delaying treatments (e.g. water, sucrose feeding, controlled atmosphere). The gene is also expressed in leaves during aging-related and harvest-induced senescence. Analysis of protein bands that cross-react with antibodies raised to the bacterial BoCP5 fusion protein, revealed prominent immunoreactive bands at ca. 26, 28, 31, and 38 kD in floret tissue. The 31 kD band was absent in protein extracts from leaf tissue. Agrobacterium-mediated transformation was used to produce transgenic broccoli plants with down-regulated BoCP5. A reduction in the postharvest expression of BoCP5 in floret tissue was achieved for four transgenic lines in the current study. In three of these lines postharvest floret senescence (yellowing) was delayed, and florets contained significantly greater chlorophyll levels during postharvest storage at 20 degrees C than wild-type plants. Line 4 showed the greatest down-regulation of BoCP5, and in this line postharvest protease activity remained at pre-harvest levels, and the yield of soluble proteins extracted from florets after harvest was significantly greater than that of wild-type tissue. Publication Types: Research Support, Non-U.S. Gov't PMID: 15988561 [PubMed - indexed for MEDLINE] 977: Toxicol Appl Pharmacol. 2005 Sep 1;207(2 Suppl):19-27. What determines the acceptability of genetically modified food that can improve human nutrition? Purchase IF. University of Manchester, Oxford Road, Manchester M13 9PT, UK. ifhp@chadzombe.u-net.com It has been predicted that by 2025 there will be an annual shortfall of cereals for feeding the human population of 68.5 million tones. One possible solution is the use of genetically modified (GM) crops, which are already grown extensively (59 million ha of GM crops were planted in 2002) in the USA, South America, Africa and China. Nevertheless, there is considerable disagreement about the advisability of using such crops, particularly in Europe. Obviously, the safety of the food derived from the GM crops is a primary consideration. Safety assessment relies on establishing that the food is substantially equivalent to its non-GM counterpart and specific testing for allergenicity of proteins and toxicity of metabolites and the whole food. There appears to be international agreement on the principles of safety assessment. Safety to the environment is equally important, but will not be covered in this presentation. The public's perception of the risk of new technology is critical to its acceptance. Perception of risk, in turn, depends on the credibility of the source of the information and trust in the regulatory process. In many countries, the public appears to have lost its trust in the scientists and government dealing with GM food, making the acceptability of GM crops uncertain. Of equal importance are the socio-economic factors that impinge on the viability of GM produce. These include intellectual property protection, trade liberalization (through subsidy and tariff barriers in developed countries) and the intensity of bio safety regulations. The socio-economic interests of developed and developing countries may diverge and may even be contradictory in any one country. Acceptance of GM crops will thus depend on detailed issues surrounding particular crops and economies. Publication Types: Lectures PMID: 15982686 [PubMed - indexed for MEDLINE] 978: Nutr Rev. 2005 May;63(5):171-81. Science, law, and politics in the Food and Drug Administration's genetically engineered foods policy: FDA's 1992 policy statement. Pelletier DL. Division of Nutritional Sciences, Cornell University, 378 MVR Hall, Ithaca, NY 14853, USA. dlp5@cornell.edu The US Food and Drug Administration's (FDA's) 1992 policy statement was developed in the context of critical gaps in scientific knowledge concerning the compositional effects of genetic transformation and severe limitations in methods for safety testing. FDA acknowledged that pleiotropy and insertional mutagenesis may cause unintended changes, but it was unknown whether this happens to a greater extent in genetic engineering compared with traditional breeding. Moreover, the agency was not able to identify methods by which producers could screen for unintended allergens and toxicants. Despite these uncertainties, FDA granted genetically engineered foods the presumption of GRAS (Generally Recognized As Safe) and recommended that producers use voluntary consultations before marketing them. Publication Types: Review PMID: 15971412 [PubMed - indexed for MEDLINE] 979: Sci Cult (Lond). 2002 Dec;11(4):459-79. Selling the life sciences: promises of a better future in biotechnology advertisements. Hellstein I. Universiteit van Amsterdam, ASCoR (Amsterdam School of Communications Research), Kloveniersburgwal 48, NL-1012CX, Amsterdam, The Netherlands. hellsten@pscw.uva.nl Publication Types: Research Support, Non-U.S. Gov't PMID: 15971365 [PubMed - indexed for MEDLINE] 980: Sci Cult (Lond). 2001 Dec;10(4):445-81. Creating public alienation: expert cultures of risk and ethics on GMOs. Wynne B. CSEC, Lancaster University, Lancaster LA1, UK. B.Wynne@Lancaster.ac.uk PMID: 15971363 [PubMed - indexed for MEDLINE] 981: Arh Hig Rada Toksikol. 2005 Jun;56(2):185-93. Food safety evaluation of crops produced through genetic engineering--how to reduce unintended effects? Jelenić S. Department of Molecular Biology, Faculty of Science, University of Zagreb, Zagreb, Croatia. sjelen@zg.biol.pmf.hr Scientists started applying genetic engineering techniques to improve crops two decades ago; about 70 varieties obtained via genetic engineering have been approved to date. Although genetic engineering offers the most precise and controllable genetic modification of crops in entire history of plant improvement, the site of insertion of a desirable gene cannot be predicted during the application of this technology. As a consequence, unintended effects might occur due to activation or silencing of genes, giving rise to allergic reactions or toxicity. Therefore, extensive chemical, biochemical and nutritional analyses are performed on each new genetically engineered variety. Since the unintended effects may be predictable on the basis of what is known about the insertion place of the transgenic DNA, an important aim of plant biotechnology is to define techniques for the insertion of transgene into the predetermined chromosomal position (gene targeting). Although gene targeting cannot be applied routinely in crop plants, given the recent advances, that goal may be reached in the near future. PMID: 15968835 [PubMed - indexed for MEDLINE] 982: Bull Entomol Res. 2005 Jun;95(3):243-7. Laboratory studies of the effects of reduced prey choice caused by Bt plants on a predatory insect. Schuler TH, Clark AJ, Clark SJ, Poppy GM, Stewart CN Jr, Denholm I. Plant and Invertebrate Ecology Division, Rothamsted Research, Harpenden, AL5 2JQ, UK. Tanja.Schuler@bbsrc.ac.uk Crops transformed to express Bacillus thuringiensis (Bt) toxins can cause close to 100% mortality of certain target pest species. This study assessed the effect of target pest reduction on the predatory insect Chrysoperla carnea (Stephens) in the presence of alternative prey. Numbers of lacewings recovered from Bt oilseed rape (cultivar Oscar, event O52) did not differ significantly from numbers of lacewings recovered from conventional oilseed rape in cage experiments with the target pest Plutella xylostella (Linnaeus) and the non-target pest Myzus persicae (Sulzer) when aphid densities were high. However, significantly fewer lacewings were recovered from Bt plants as aphid densities were lowered. Lacewing weights were not affected by plant type. Publication Types: Comparative Study Research Support, Non-U.S. Gov't PMID: 15960878 [PubMed - indexed for MEDLINE] 983: J Fish Dis. 2005 Jun;28(6):317-30. An examination of the intestinal tract of Atlantic salmon, Salmo salar L., parr fed different varieties of soy and maize. Sanden M, Berntssen MH, Krogdahl A, Hemre GI, Bakke-McKellep AM. National Institute of Nutrition and Seafood Research (NIFES), Bergen, Norway. monica.sanden@nifes.no This study was conducted to investigate the long-term effects of feeding plant products from both traditional breeding and from biotechnology on intestinal somatic indices, histology and cell proliferation in first-feeding Atlantic salmon, Salmo salar L. (initial weight 0.21 +/- 0.02 g). A standard fishmeal diet (standard fishmeal) was formulated to contain fishmeal as the sole protein source and suprex maize as the main starch source. Six experimental diets were then developed: two in which some of the fishmeal was replaced with commercially available, genetically modified Roundup Ready full-fat soybean meal (GM-soy) or commercially available, non-GM full-fat soybean meal (nGM-soy) at a level of 12.5% of the total diet, and four diets in which the suprex maize was replaced with two lines of GM-maize (Dekalb 1; D1 and Pioneer 1; P1), both products of event MON810, and their half-sibling non-GM counterparts (Dekalb 2; D2 and Pioneer 2; P2), at a level of 12.1% of total diet. Each diet was fed to fish in triplicate tanks and the experiment lasted for 8 months, during which the fish reached a final weight of 101-116 g. There was no significant effect of diet on the intestinal indices, nor were histological changes observed in the pyloric caeca or mid intestine. In the distal intestine, one of nine sampled fish fed nGM-soy showed moderate changes, two of nine sampled fish fed GM-soy showed changes, one with moderate and one with severe changes, and two of nine fish fed nGM-maize D2 had moderate changes. Using a monoclonal antibody against proliferating cell nuclear antigen (PCNA), cell proliferative responses to the experimental diets were assessed. In fish fed both soy diets, a significantly higher (P < 0.05) cell proliferation response was observed in the distal intestine concomitant with an increased localization of PCNA positive cells along the whole distal intestinal folds. The PCNA response among the nGM-soy group was significantly higher compared with all the other diet groups. In contrast, for fish exposed to dietary maize (type D) compared with fish fed the standard fishmeal, the soy-diets (GM-soy and nGM-soy) and maize (type P), a significantly lower (P < 0.05) cell proliferation response was observed in the distal intestine. Results indicated that the GM plant products investigated in this study, at about 12% inclusion level, were as safe as commercially available non-GM products, at least in terms of their effect on indices and histological parameters of the Atlantic salmon intestinal tract. Publication Types: Comparative Study PMID: 15960655 [PubMed - indexed for MEDLINE] 984: Ann N Y Acad Sci. 2005 May;1041:77-81. Expression of porcine prorelaxin in transgenic tobacco. Buswell S, Medina-Bolivar F, Chen Q, Van Cott K, Zhang C. Department of Biological Systems Engineering, Virginia Polytechnic Institute and State University, Blacksburg, Virginia 24061, USA. In vitro studies demonstrate that porcine relaxin may possess various therapeutic effects. In this study, we explore the possibility of expressing porcine relaxin in transgenic tobacco. Tobacco was selected because it is a non-food, non-feed crop, and recombinant protein production can readily be scaled up. The cDNA of porcine preprorelaxin was under the regulation of two different constitutive promoters. DNA analysis by polymerase chain reaction verified that all transgenic plants contained the correct size of gene insert. Preliminary studies showed the presence of putative prorelaxin bands in both silver-stained SDS-PAGE and western blot. The results also indicated that tobacco-produced prorelaxin may not be properly processed to yield the mature relaxin. PMID: 15956689 [PubMed - indexed for MEDLINE] 985: Meded Rijksuniv Gent Fak Landbouwkd Toegep Biol Wet. 2001;66(3b):631-4. Extraction and PCR analysis of soy DNA in chocolate. Supli K, Gryson N, Messens K, De Loose M, Dewettinck K. AgriFing, Centre for Applied Research and Services, Hogeschool Gent, Voskenslaan 270, B-9000 Gent, Belgium. The manner in which to discriminate between genetically modified (GM) versus non-modified foodstuffs is based on the presence of newly introduced genes at the protein or DNA level. Current available methods are almost exclusively based on the polymerase chain reaction (PCR). These methods consist of three steps: DNA isolation, amplification of the desired DNA fragment and visualisation of the obtained amplification products. The first and crucial step is the DNA isolation. In this study three different methods are described for the isolation of DNA from chocolate, two of which are commercial kits. The results indicate that both kits, in contrast with the non-commercial method, are suitable for the isolation of DNA from chocolate, provided that the adapted PCR conditions are applied. Publication Types: Research Support, Non-U.S. Gov't PMID: 15954670 [PubMed - indexed for MEDLINE] 986: Meded Rijksuniv Gent Fak Landbouwkd Toegep Biol Wet. 2001;66(3b):469-72. Use of cloned DNA fragments as reference materials for event specific quantification of genetically modified organisms (GMOs). Taverniers I, Van Bockstaele E, De Loose M. Department for Plant Genetics and Breeding, CLO-Gent, Caritasstraat 21, B-9090 Melle, Belgium. For the quantification of genetically modified organisms (GMOs) in foods and feeds, real-time PCR is currently the most widely applied technique. To obtain a % of GMO, a GMO-specific target sequence is quantified relatively to a species-specific sequence. The correctness and reliability of the obtained quantitative results fully depend on the reference materials used as standards for setting up external calibration curves. We introduced a completely new type of standards for quantification of GMOs, based on cloned plasmid DNA solutions with well-known amounts of the sequences of interest, expressed as copy numbers. Moreover, the junction sequence between inserted DNA and plant DNA was used as 'unique identifier'. In this study, the model was applied for Roundup Ready soybean. PMID: 15954640 [PubMed - indexed for MEDLINE] 987: Meded Rijksuniv Gent Fak Landbouwkd Toegep Biol Wet. 2001;66(3b):443-6. Wheat transformation with glutenin gene. Delporte F, Jérouville B, Mélard A, Kutten L, Baleux R, Iuliano M, Schellingen D, Jacquemin JM. Centre de Recherches agronomiques- Département de Biotechnologie, Chaussée de Charleroi 234, 5030 Gembloux, Belgium. The rheological properties of wheat grains are associated with the composition of the starchy endosperm in high molecular weight (HMW) glutenin proteins. The HMW glutenin 1xDy12 subunit gene was co introduced with the screenable bar and the reporter gus marker genes in the commercial spring wheat Minaret cultivar (cv). The gene of interest and the marker genes were carried by two separated plasmids, the pBS10BH1 and the pAhC25 respectively. Seven days old calli initiated from immature embryos were bombarded by use of the PDS-1000/He device. A number of different bombardment and culture conditions were tested. These parameters were evaluated on the basis of GUS transient expression. Among those a 4 degrees C cold pre-treatment of the spikes before immature embryos were excised, the culture medium incorporating activated charcoal, silver nitrate and glucose yielded higher GUS transient expression rate. The selective agent bialaphos was maintained at various stages during culture from induction of somatic embryogenesis to rooting of regenerated plantlets. 137 bialaphos resistant plants were obtained among which 109 were carried to maturity. Transgenic plants were characterized by PCR, Southern and SDS-PAGE analysis of the glutenin content. PMID: 15954633 [PubMed - indexed for MEDLINE] 988: Meded Rijksuniv Gent Fak Landbouwkd Toegep Biol Wet. 2001;66(3b):435-6. Improvement of sorghum transformation efficiency for increasing nutritional quality. Chakraborty R, Jacobs M, Angenon G. Laboratory of Plant Genetics, Institute of Molecular Biology and Biotechnology, Vrije Universiteit Brussel, Paadenstraat 65, B-1640 Genesius Rode, Belgium. PMID: 15954630 [PubMed - indexed for MEDLINE] 989: Meded Rijksuniv Gent Fak Landbouwkd Toegep Biol Wet. 2001;66(3b):417-24. Refining of soy bean oil: detection of DNA. Gryson N, Ronsse F, Messens K, De Loose M, Verleyen T, Dewettinck K. AgriFing, Centre for Applied Research and Services, Hogeschool Gent, Voskenslaan 270, B-9000 Gent, Belgium. Publication Types: Research Support, Non-U.S. Gov't PMID: 15954627 [PubMed - indexed for MEDLINE] 990: Meded Rijksuniv Gent Fak Landbouwkd Toegep Biol Wet. 2001;66(3a):31-7. Biotechnology and the consumer. Debeuckelaere W. Test-Aankoop, Hollandstraat 13, 1060 Brussel, Belgium. PMID: 15954560 [PubMed - indexed for MEDLINE] 991: Meded Rijksuniv Gent Fak Landbouwkd Toegep Biol Wet. 2001;66(4):335-9. Consumers and GM food: a divergent relation. Verdurme A. Ghent University, Faculty of Agricultural and Applied Biological Sciences Department of Agricultural Economics, Division of Agro-marketing, Coupure Links 653, 9000 Gent, Belgium. Annelies.Verdurme@rug.ac.be Publication Types: Research Support, Non-U.S. Gov't PMID: 15954315 [PubMed - indexed for MEDLINE] 992: Trends Biotechnol. 2005 Aug;23(8):386-7. Comment on: Trends Biotechnol. 2004 Mar;22(3):107-9. Consumer acceptance of biotechnology and the role of second generation technologies in the USA and Europe. Lusk JL, Rozan A. Publication Types: Comment Letter PMID: 15953650 [PubMed - indexed for MEDLINE] 993: Wei Sheng Yan Jiu. 2005 Mar;34(2):244-8. [Strategies for safety assessment of genetically modified crops: current and future development] [Article in Chinese] Zhuo Q, Yang XG. National Institute of Nutrition and Food Safety, Chinese Centre for Disease Control and Prevention, Beijing 100050, China. Gene recombinant technologies supply agriculture product with great vitality. But the risk of genetically modified crops cannot be ignored. The international organizations such as WHO, FAO and OECD have reached common agreement: the safety of transgenic crops should be thoroughly evaluated based on "substantial equivalence"principle. The relevant strategies including: substantial equivalent analysis, toxic tests, protein allergenic study, nutritional assessment, etc. With the development of new technologies, the approaches of genomic, proteomics, metabolomics would be applied to detect the unintended effects. The EU recently adopted legislation on the cultivation GM crops requiring the post market surveillance for any unanticipated adverse effects in the long term. In conclusion, the strategies of the safety assessment of GM crop are very strict and likely development. Publication Types: English Abstract Research Support, Non-U.S. Gov't Review PMID: 15952675 [PubMed - indexed for MEDLINE] 994: Nat Biotechnol. 2005 Aug;23(8):1013-7. Epub 2005 Jun 12. Stepwise engineering to produce high yields of very long-chain polyunsaturated fatty acids in plants. Wu G, Truksa M, Datla N, Vrinten P, Bauer J, Zank T, Cirpus P, Heinz E, Qiu X. Bioriginal Food & Science Corporation, 110 Gymnasium Place, Saskatoon, Saskatchewan, Canada S7N 0W9. Very long chain polyunsaturated fatty acids (VLCPUFAs) such as arachidonic acid (AA), eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) are valuable commodities that provide important human health benefits. We report the transgenic production of significant amounts of AA and EPA in Brassica juncea seeds via a stepwise metabolic engineering strategy. Using a series of transformations with increasing numbers of transgenes, we demonstrate the incremental production of VLCPUFAs, achieving AA levels of up to 25% and EPA levels of up to 15% of total seed fatty acids. Both fatty acids were almost exclusively found in triacylglycerols, with AA located preferentially at sn-2 and sn-3 positions and EPA distributed almost equally at all three positions. Moreover, we reconstituted the DHA biosynthetic pathway in plant seeds, demonstrating the practical feasibility of large-scale production of this important omega-3 fatty acid in oilseed crops. Publication Types: Evaluation Studies PMID: 15951804 [PubMed - indexed for MEDLINE] 995: Vet Parasitol. 2005 Aug 10;131(3-4):207-11. First isolation and molecular characterization of Toxoplasma gondii from finishing pigs from São Paulo State, Brazil. de A Dos Santos CB, de Carvalho AC, Ragozo AM, Soares RM, Amaku M, Yai LE, Dubey JP, Gennari SM. Departamento de Medicina Veterinária Preventiva, Faculdade de Ciências Agrárias e Veterinárias da Universidade Estadual Paulista, Jaboticabal, SP, Brazil. Toxoplasma gondii infection is widely prevalent in humans in Brazil. Among the food animals, pigs are considered the most important meat source of T. gondii for infection in humans. In the present study, we report the first isolation of viable T. gondii from finishing pigs in Brazil. Antibodies to T. gondii were found in 49 (17%) of 286 pigs prior slaughter using the modified agglutination test (MAT) at a serum dilution of 1:25. Attempts were made to isolate T. gondii from 28 seropositive pigs. Samples of heart, brain, and tongue from each pig were pooled, digested in acid pepsin, and bioassayed in five mice per pig. Viable T. gondii was isolated from seven pigs; all isolates were lethal for mice. Restriction fragment length polymorphism on products of SAG2 locus amplified by PCR revealed that two isolates were Type I and five were Type III. The results indicate that phenotypically and genetically T. gondii isolates from pigs from Brazil are distinct from isolates of T. gondii from pigs in the USA. PMID: 15951111 [PubMed - indexed for MEDLINE] 996: Int Arch Allergy Immunol. 2005 Jun;137(2):167-80. Epub 2005 Jun 8. Comment in: Int Arch Allergy Immunol. 2005 Jun;137(2):151-2. Suggestions for the assessment of the allergenic potential of genetically modified organisms. Spök A, Gaugitsch H, Laffer S, Pauli G, Saito H, Sampson H, Sibanda E, Thomas W, van Hage M, Valenta R. Inter-University Research Centre for Technology, Work, and Culture, Graz, Austria. The prevalence of allergic diseases has been increasing continuously and, accordingly, there is a great desire to evaluate the allergenic potential of components in our daily environment (e.g., food). Although there is almost no scientific evidence that genetically modified organisms (GMOs) exhibit increased allergenicity compared with the corresponding wild type significant concerns have been raised regarding this matter. In principle, it is possible that the allergenic potential of GMOs may be increased due to the introduction of potential foreign allergens, to potentially upregulated expression of allergenic components caused by the modification of the wild type organism or to different means of exposure. According to the current practice, the proteins to be introduced into a GMO are evaluated for their physiochemical properties, sequence homology with known allergens and occasionally regarding their allergenic activity. We discuss why these current rules and procedures cannot predict or exclude the allergenicity of a given GMO with certainty. As an alternative we suggest to improve the current evaluation by an experimental comparison of the wild-type organism with the whole GMO regarding their potential to elicit reactions in allergic individuals and to induce de novo sensitizations. We also recommend that the suggested assessment procedures be equally applied to GMOs as well as to natural cultivars in order to establish effective measures for allergy prevention. Publication Types: Research Support, Non-U.S. Gov't PMID: 15947472 [PubMed - indexed for MEDLINE] 997: Int Arch Allergy Immunol. 2005 Jun;137(2):153-66. Epub 2005 Jun 8. Comment in: Int Arch Allergy Immunol. 2005 Jun;137(2):151-2. Assessing genetically modified crops to minimize the risk of increased food allergy: a review. Goodman RE, Hefle SL, Taylor SL, van Ree R. Food Allergy Research and Resource Program, University of Nebraska, Lincoln, NE 68583-0955, USA. rgoodman2@unlnotes.unl.edu The first genetically modified (GM) crops approved for food use (tomato and soybean) were evaluated for safety by the United States Food and Drug Administration prior to commercial production. Among other factors, those products and all additional GM crops that have been grown commercially have been evaluated for potential increases in allergenic properties using methods that are consistent with the current understanding of food allergens and knowledge regarding the prediction of allergenic activity. Although there have been refinements, the key aspects of the evaluation have not changed. The allergenic properties of the gene donor and the host (recipient) organisms are considered in determining the appropriate testing strategy. The amino acid sequence of the encoded protein is compared to all known allergens to determine whether the protein is a known allergen or is sufficiently similar to any known allergen to indicate an increased probability of allergic cross-reactivity. Stability of the protein in the presence of acid with the stomach protease pepsin is tested as a risk factor for food allergenicity. In vitro or in vivo human IgE binding are tested when appropriate, if the gene donor is an allergen or the sequence of the protein is similar to an allergen. Serum donors and skin test subjects are selected based on their proven allergic responses to the gene donor or to material containing the allergen that was matched in sequence. While some scientists and regulators have suggested using animal models, performing broadly targeted serum IgE testing or extensive pre- or post-market clinical tests, current evidence does not support these tests as being predictive or practical. Based on the evidence to date, the current assessment process has worked well to prevent the unintended introduction of allergens in commercial GM crops. Publication Types: Review PMID: 15947471 [PubMed - indexed for MEDLINE] 998: Int Arch Allergy Immunol. 2005 Jun;137(2):151-2. Epub 2005 Jun 8. Comment on: Int Arch Allergy Immunol. 2005 Jun;137(2):153-66. Int Arch Allergy Immunol. 2005 Jun;137(2):167-80. Can we predict or avoid the allergenic potential of genetically modified organisms? Obermeyer G, Ferreira F. Publication Types: Comment Editorial PMID: 15947470 [PubMed - indexed for MEDLINE] 999: Nat Biotechnol. 2005 Jun;23(6):636. Beer giant blindsides Ventria's pharmacrop. Fox JL. Publication Types: News PMID: 15940219 [PubMed - indexed for MEDLINE] 1000: Biosens Bioelectron. 2006 Jan 15;21(7):1069-76. Epub 2005 Jun 2. Nanoparticle-based DNA biosensor for visual detection of genetically modified organisms. Kalogianni DP, Koraki T, Christopoulos TK, Ioannou PC. Department of Chemistry, University of Patras, Patras 26500, Greece. Although screening of raw ingredients and food products for genetically modified organisms (GMO) may be accomplished by detecting either the exogenous DNA or the novel protein, DNA is the preferred analyte because of its superior stability during food processing. The development of DNA biosensors is of increasing importance due to the growing demand for rapid and reliable methods for GMO detection. We report the first DNA biosensor in a dry-reagent dipstick configuration for visual detection and confirmation of GMO-related sequences by hybridization within minutes. The sensor is disposable and does not require special instrumentation. It detects the 35S promoter and nopaline synthase (NOS) terminator sequences that are present in the majority of transgenic plants. The target sequences are amplified by the polymerase chain reaction (PCR) and hybridized (7min) with probes bearing oligo(dA) tail. The biotinylated product is applied to the sensor followed by immersion in the appropriate buffer. Migration of the buffer rehydrates gold nanoparticles conjugated to oligo(dT), which hybridize with the oligo(dA) tails. The hybrids are captured by immobilized streptavidin at the test zone of the sensor giving a characteristic red line due to the accumulation of the nanoparticles. The excess of nanoparticle conjugates are captured at the control zone by immobilized oligo(dA) strands. Amplified 35S or NOS DNA is detectable at 0.16nM. Soybean powder certified reference material with 0.1% GMO content is clearly detectable after 35 and 40 amplification cycles for 35S and NOS sequence, respectively. The sensor was also applied to real samples from various sources. Publication Types: Validation Studies PMID: 15935636 [PubMed - indexed for MEDLINE] 1001: FEMS Microbiol Rev. 2005 Aug;29(3):611-24. The long and winding road from the research laboratory to industrial applications of lactic acid bacteria. Pedersen MB, Iversen SL, Sørensen KI, Johansen E. Research, Development and Application, Chr. Hansen A/S, 10-12 Bøge Allé, DK2970 Hørsholm, Denmark. Research innovations are constantly occurring in universities, research institutions and industrial research laboratories. These are reported in the scientific literature and presented to the scientific community in various congresses and symposia as well as through direct contacts and collaborations. Conversion of these research results to industrially useful innovations is, however, considerably more complex than generally appreciated. The long and winding road from the research laboratory to industrial applications will be illustrated with two recent examples from Chr. Hansen A/S: the implementation in industrial scale of a new production technology based on respiration by Lactococcus lactis and the introduction to the market of L. lactis strains constructed using recombinant DNA technology. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 15935510 [PubMed - indexed for MEDLINE] 1002: Eur J Neurosci. 2005 May;21(9):2600-4. Mice lacking dopamine D1 receptors express normal lithium chloride-induced conditioned taste aversion for salt but not sucrose. Cannon CM, Scannell CA, Palmiter RD. Department of Biochemistry and Howard Hughes Medical Institute, University of Washington, Seattle, WA 98195-7370, USA. caesia@u.washington.edu Conditioned taste aversion (CTA), is a form of Pavlovian learning wherein a novel flavour is powerfully associated with subsequent feelings of illness, and is afterwards avoided. In rats, pharmacological blockade of dopamine D1 receptors has been reported to prevent the expression of a CTA to the sweet taste of sucrose or saccharine. We used genetically modified mice to determine whether dopamine D1 receptors are necessary for the expression of a CTA. Food-deprived mice lacking the dopamine D1 receptor (D1r-/-) did not express a LiCl-induced (125 or 254 mg/kg) CTA to the sweet taste of 0.5 m sucrose, in agreement with previous pharmacological studies. However, water-deprived D1r-/- mice did express normal LiCl-induced (40, 150 and 254 mg/kg) CTA to a salty taste (0.2 m NaCl). Our results suggest that activation of D1 receptors might contribute to the strength of an aversive gustatory association, but might not be required for the formation of a CTA in general. Publication Types: Research Support, N.I.H., Extramural Research Support, U.S. Gov't, P.H.S. PMID: 15932618 [PubMed - indexed for MEDLINE] 1003: Nature. 2005 Jun 2;435(7042):561. Comment on: Nature. 2005 Apr 14;434(7035):807. Ampicillin threat leads to wider transgene concern. Azeez G. Publication Types: Comment Letter PMID: 15931193 [PubMed - indexed for MEDLINE] 1004: Nature. 2005 Jun 2;435(7042):561. Comment in: Nature. 2005 Jul 21;436(7049):328. Nature. 2005 Jul 21;436(7049):328. Comment on: Nature. 2005 Mar 31;434(7033):559. Activists should accept mainstream view of GM. Dennis DT. Publication Types: Comment Letter PMID: 15931192 [PubMed - indexed for MEDLINE] 1005: Environ Health Perspect. 2005 Jun;113(6):716-20. Comment in: Environ Health Perspect. 2005 Oct;113(10):A657-8; author reply A658. Differential effects of glyphosate and roundup on human placental cells and aromatase. Richard S, Moslemi S, Sipahutar H, Benachour N, Seralini GE. Laboratoire de Biochimie et Biologie Moleculaire, USC-INCRA, Université de Caen, Caen, France. Roundup is a glyphosate-based herbicide used worldwide, including on most genetically modified plants that have been designed to tolerate it. Its residues may thus enter the food chain, and glyphosate is found as a contaminant in rivers. Some agricultural workers using glyphosate have pregnancy problems, but its mechanism of action in mammals is questioned. Here we show that glyphosate is toxic to human placental JEG3 cells within 18 hr with concentrations lower than those found with agricultural use, and this effect increases with concentration and time or in the presence of Roundup adjuvants. Surprisingly, Roundup is always more toxic than its active ingredient. We tested the effects of glyphosate and Roundup at lower nontoxic concentrations on aromatase, the enzyme responsible for estrogen synthesis. The glyphosate-based herbicide disrupts aromatase activity and mRNA levels and interacts with the active site of the purified enzyme, but the effects of glyphosate are facilitated by the Roundup formulation in microsomes or in cell culture. We conclude that endocrine and toxic effects of Roundup, not just glyphosate, can be observed in mammals. We suggest that the presence of Roundup adjuvants enhances glyphosate bioavailability and/or bioaccumulation. Publication Types: Comparative Study Research Support, Non-U.S. Gov't PMID: 15929894 [PubMed - indexed for MEDLINE] 1006: J Environ Manage. 2005 Aug;76(3):210-20. Regulating insect resistance management: the case of non-Bt corn refuges in the US. Bourguet D, Desquilbet M, Lemarié S. Centre de Biologie et de Gestion des Populations, UMR INRA/IRD/CIRAD/ENSA.M, Campus International de Baillarguet, CS 30 016, 34 988 Montferrier/Lez, France. bourguet@ensam.inra.fr In this paper, we analyze the insect resistance management (IRM) plan put in place by the US Environmental Protection Agency (EPA) to delay the evolution of resistance to Bt corn in natural populations of the European corn borer, Ostrinia nubilalis Hübner. This IRM plan is the most impressive mandatory IRM system ever developed. It forms a coherent whole and includes mandatory refuges, actions to increase growers' compliance and a program for monitoring the evolution of resistance. However, our analysis suggests that two components of this IRM plan are not entirely satisfactory: growers' compliance and monitoring of the evolution of resistance. Moreover, the implementation of these two components of IRM has been required of the registrants, whose incentives for IRM are probably lower than the social optimum. Our analysis suggests that alternatives to the IRM plan currently in place could improve these two components. Publication Types: Comparative Study Research Support, Non-U.S. Gov't Review PMID: 15922504 [PubMed - indexed for MEDLINE] 1007: Trends Biotechnol. 2005 Jun;23(6):308-15. Tuning the pores: towards engineering plants for improved water use efficiency. Chaerle L, Saibo N, Van Der Straeten D. Unit Plant Hormone Signaling and Bio-imaging, Ghent University, Ledeganckstraat 35, B-9000 Gent, Belgium. The management of limited fresh water resources is a major challenge facing society in the 21st century. The agricultural sector accounts for more than two-thirds of human water withdrawal and is therefore a prime area to implement a more rational water use. Environmental stresses are a major factor limiting stable food production. Given the growing shortage of available water for crops this will be an emerging factor in international agricultural economy. The most environmentally friendly and durable solution to the problem of water shortage is to complement more efficient irrigation approaches with crops with optimal water use efficiency, achieved either through genetic engineering or conventional breeding, combined with high yields. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 15922083 [PubMed - indexed for MEDLINE] 1008: Endocrinology. 2005 Sep;146(9):3800-8. Epub 2005 May 26. Decreased gonadotropin-releasing hormone neuronal activity is associated with decreased fertility and dysregulation of food intake in the female GPR-4 transgenic rat. Gomez F, la Fleur SE, Weiner RI, Dallman MF, El Majdoubi M. Department of Obstetrics, 513 Parnassus Avenue, HSW1475, Box 0556, University of California, San Francisco, California 94143, USA. Expression of a cAMP-specific phosphodiesterase in GnRH neurons in the GPR-4 transgenic rat resulted in decreased LH levels and pulse frequency and diminished fertility. We have characterized changes in fertility, adiposity, and reproductive and metabolic hormones with age. Although LH levels were decreased in 3-, 6-, and 9-month-old GPR-4 females relative to wild-type (WT) controls, GPR-4 females did not become anovulatory until 6 months of age. No differences were observed in FSH, estradiol, or androstenedione levels in 3-, 6-, or 9-month-old GPR-4 and WT females. At 9 months of age, GPR-4 females had significantly increased abdominal and sc fat depot weights that were associated with increased leptin and insulin levels not observed in WT females. We tested the hypothesis that metabolic changes observed at 9 months of age were the result of dysregulation of the mechanisms controlling energy balance. Two-month-old female GPR-4 rats placed on a high-energy diet gained weight at a rate significantly greater than WT females and, after 24 d, developed the same metabolic phenotype observed in 9-month-old GRP-4 females (increased abdominal and sc fat associated with elevated leptin and insulin concentrations). Overeating did not correlate with changes in estradiol or androstenedione levels. We conclude that decreased GnRH neuronal activity is closely associated with decreased reproductive function and dysregulation of food intake. Publication Types: Research Support, N.I.H., Extramural Research Support, U.S. Gov't, P.H.S. PMID: 15919747 [PubMed - indexed for MEDLINE] 1009: Lancet Infect Dis. 2005 Jun;5(6):330. Scoring at the wrong end. Dixon B. dixonadams@blueyonder.co.uk PMID: 15919619 [PubMed - indexed for MEDLINE] 1010: Lancet. 2005 May 21-27;365(9473):1746. Comment in: Lancet. 2005 Jul 16-22;366(9481):202-3. Biotech quick-fixes will not end hunger in China. [No authors listed] Publication Types: Editorial PMID: 15915561 [PubMed - indexed for MEDLINE] 1011: Biosci Biotechnol Biochem. 2005 May;69(5):966-71. Avidin expressed in transgenic rice confers resistance to the stored-product insect pests Tribolium confusum and Sitotroga cerealella. Yoza K, Imamura T, Kramer KJ, Morgan TD, Nakamura S, Akiyama K, Kawasaki S, Takaiwa F, Ohtsubo K. National Food Research Institute, Ibaraki, Japan. yozako@affrc.go.jp Rice (Oryza sativa var. Nipponbare) was transformed with an artificial avidin gene. The features of this construct are as follows: (1) a signal peptide sequence derived from barley alpha amylase was added at the N-terminal region, (2) codon usage of the gene was optimized for rice, and (3) the gene was driven by rice glutelin GluB-1, an endosperm-specific promoter. Avidin was produced in the grain of the transgenic rice but not in the leaves. The concentration of avidin in the kernels was about 1,800 ppm. All larvae of the confused flour beetle (Tribolium confusum) and Angoumois grain moth (Sitotroga cerealella) died when fed transgenic avidin rice powder or kernels, respectively, whereas most of the test insects developed into adults when they were fed a nontransgenic rice control diet. Avidin extracted from the transgenic rice kernel lost most biotin-binding activity after 5 min heating at 95 degrees C. Publication Types: Research Support, Non-U.S. Gov't PMID: 15914917 [PubMed - indexed for MEDLINE] 1012: J Agric Food Chem. 2005 Jun 1;53(11):4315-21. Application of immunoaffinity column as cleanup tool for an enzyme linked immunosorbent assay of phosphinothricin-N-acetyltransferase detection in genetically modified maize and rape. Xu W, Huang K, Zhao H, Luo Y. College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, China. We have developed a new immunoassay method to detect genetically modified (GM) maize and rape containing phosphinothricin-N-acetyltransferase (PAT). PAT encoded by Bialaphos resistance gene (bar) was highly expressed in soluble form in Escherichia coli BL21(DE3) and purified to homogeneity by Ni2+ affinity chromatography. A simple and efficient extraction and purification procedure of PAT from GM maize and rape was developed by means of the immunoaffinity column (IAC) as a cleanup tool. Purified polyclonal antibodies against PAT was produced and coupled covalently to CNBr-activated Sepharose 4B. Both the binding conditions and elution protocols were optimized. The IAC was successfully employed to isolate and purify the PAT from the various tissues of GM maize (Bt11 and Bt176) and rapes (MS1/RF1 and MS8/RF3). Enzyme linked immunosorbent assay (ELISA) procedures were established further on to measure the PAT protein. GM maize cannot be differentiated from non-GM maize by ELISA. But IAC-ELISA allowed 0.5% GMOs to be detected in MS1/RF1 and MS8/RF3 and 10% GMOs to be detected in Bt11 and Bt176, which makes this method an acceptable method to access PAT protein in GM rapes and maize. Publication Types: Research Support, Non-U.S. Gov't PMID: 15913288 [PubMed - indexed for MEDLINE] 1013: Rev Esp Salud Publica. 2005 Mar-Apr;79(2):271-82. [Genetically modified organisms: a new threat to food safety] [Article in Spanish] Spendeler L. Amigos de la Tierra España, Madrid. transgenicos@tierra.org This article analyzes all of the food safety-related aspects related to the use of genetically modified organisms into agriculture and food. A discussion is provided as to the uncertainties related to the insertion of foreign genes into organisms, providing examples of unforeseen, undesirable effects and of instabilities of the organisms thus artificially fabricated. Data is then provided from both official agencies as well as existing literature questioning the accuracy and reliability of the risk analyses as to these organisms being harmless to health and discusses the almost total lack of scientific studies analyzing the health safety/dangerousness of transgenic foods. Given all these unknowns, other factors must be taken into account, particularly genetic contamination of the non-genetically modified crops, which is now starting to become widespread in some parts of the world. Not being able of reversing the situation in the even of problems is irresponsible. Other major aspects are the impacts on the environment (such as insects building up resistances, the loss of biodiversity, the increase in chemical products employed) with indirect repercussions on health and/or future food production. Lastly, thoughts for discussion are added concerning food safety in terms of food availability and food sovereignty, given that the transgenic seed and related agrochemicals market is currently cornered by five large-scale transnational companies. The conclusion entails an analysis of biotechnological agriculture's contribution to sustainability. Publication Types: English Abstract Review PMID: 15913060 [PubMed - indexed for MEDLINE] 1014: J Environ Sci Health B. 2005;40(3):463-73. The fate of the recombinant DNA in corn during composting. Guan J, Spencer JL, Ma BL. Ottawa Laboratory-Fallowfield, Canadian Food Inspection Agency, Ottawa, Ontario, Canada. guanj@inspection.gc.ca In order to make regulations that safeguard food and the environment, an understanding of the fate oftransgenes from genetically modified (GM) plants is of crucial importance. A compost experiment including mature transgenic corn plants and seeds of event Bt 176 (Zea mays L.) was conducted to trace the fate of the transgene cryIA(b) during the period of composting. In bin 1, shredded corn plants including seeds were composted above a layer of cow manure and samples from the corn layer were collected at intervals during a 12-month period. The samples were tested for the transgene persistence and microbial counts and also the compost was monitored for temperature. In bin 2, piles of corn seeds, surrounded by sheep manure and straw, were composted for 12 months. A method combining nested polymerase chain reaction (PCR) and southern hybridization was developed for detection of the transgene in compost. The detection sensitivity was 200 copies of the transgene per gram of dry composted corn material. Composting commenced on day 0, and the transgene was detected in specimens from bin 1 on days 0 and 7 but not on day 14 or thereafter. The transgene in corn seeds was not detectable after 12 months of composting in bin 2. Temperatures in both bins rose to about 50 degrees C within 2 weeks and remained above that temperature for about 3 months, even when the ambient temperature dropped below -20 degrees C. Extracts from compost were inoculated onto culture plates and then were incubated at 23 to 55 degrees C. Within the first 2 weeks of composting in bin 1, the counts of bacteria incubated at 55 degrees C increased from 3.5 to 7.5 log10, whereas those incubated at 23 degrees C remained at about 7.5 log10. The counts of fungi incubated at 45 degrees C increased slightly from 2.5 to 3.1 log10, but those incubated at 23 degrees C decreased from 6.3 to 3.0 log10. The rapid degradation of the transgene during composting of Bt corn plants suggested that the composting process could be used for safe disposal of transgenic plant wastes. Publication Types: Research Support, Non-U.S. Gov't PMID: 15913018 [PubMed - indexed for MEDLINE] 1015: Environ Biosafety Res. 2004 Jul-Sep;3(3):135-48. Management of herbicide-tolerant oilseed rape in Europe: a case study on minimizing vertical gene flow. Devos Y, Reheul D, de Schrijver A, Cors F, Moens W. Service of Biosafety and Biotechnology, Scientific Institute of Public Health, Juliette Wytsmanstraat 14, 1050 Brussels, Belgium. Yann.Devos@UGent.be The potential commercialization of genetically modified herbicide-tolerant (GMHT) oilseed rape in Europe raises various concerns about their potential environmental and agronomic impacts, especially those associated with the escape of transgenes. Pollen of oilseed rape can be dispersed in space, resulting in the fertilization of sympatric compatible wild relatives (e.g. Brassica rapa) and oilseed rape cultivars grown nearby (GM and/or non-GM Brassica napus). The spatial and temporal dispersal of seeds of oilseed rape may lead to feral oilseed rape populations outside the cropped areas and oilseed rape volunteers in subsequent crops in the rotation. The incorporation of a HT trait(s) may increase the fitness of the recipient plants, making them more abundant and persistent, and may result in weeds that are difficult to control by the herbicide(s) to which they are tolerant. Vertical gene flow from transgenic oilseed rape to non-GM counterparts may also have an impact on farming and supply chain management, depending on labelling thresholds for the adventitious presence of GM material in non-GM products. Given the extent of pollen and seed dispersal in oilseed rape, it is obvious that the safe and sound integration of GMHT oilseed rape in Europe may require significant on-farm and off-farm management efforts. Crucial practical measures that can reduce vertical gene flow include (1) isolating seed production of Brassica napus, (2) the use of certified seed, (3) isolating fields of GM oilseed rape, (4) harvesting at the correct crop development stage with properly adjusted combine settings, (5) ensuring maximum germination of shed seeds after harvest, (6) controlling volunteers in subsequent crops, and (7) keeping on-farm records. The implementation of the recommended practices may, however, be difficult, entailing various challenges. Publication Types: Review PMID: 15901096 [PubMed - indexed for MEDLINE] 1016: Appetite. 2005 Aug;45(1):47-50. Perceived naturalness and acceptance of genetically modified food. Tenbült P, de Vries NK, Dreezens E, Martijn C. Department of Health Education and Health Promotion, Maastricht University, P.O. Box 616, 6200 MD Maastricht, The Netherlands. p.tenbult@gvo.unimaas.nl This study examines people's acceptance of genetically modified (GM) food. Results suggest that GM acceptance depends most on how natural the genetically modified product is perceived and not directly on how natural the non-GM product is seen. A GM product that is perceived as more natural is more likely to be accepted than a GM product that is perceived as less natural. The extent to which GM affects the perceived naturalness of a product partly depends on the kind of product. PMID: 15896875 [PubMed - indexed for MEDLINE] 1017: Virus Res. 2005 Jul;111(1):93-100. Epub 2005 Apr 19. Plant biopharming of monoclonal antibodies. Ko K, Koprowski H. Biotechnology Foundation Laboratories at Thomas Jefferson University, 1020 Locust Street, Room M85 JAH, Philadelphia, PA 19107, USA. Recent advances in molecular biology and plant biotechnology have shifted the concept of growing crops as a food source to serving as a bioreactor for the production of therapeutic recombinant proteins. Plants are potential biopharming factories because they are capable of producing unlimited numbers and amounts of recombinant proteins safely and inexpensively. In the last two decades, plant production systems have been developed for monoclonal antibody production, which has been useful in passive immunization of viral or bacterial diseases. Recently, a recombinant monoclonal antibody for rabies prophylaxis was produced in transgenic plants. Rabies virus epidemics remain still problematic throughout the world, and adequate treatment has been hampered by the worldwide shortage and high cost of prophylactic antibodies such as HRIG. Successful mass production of this monoclonal antibody in plants might help to overcome these problems. An effective plant production system for recombinant biologicals requires the appropriate heterologous plant expression system, the optimal combination of gene expression regulatory elements, control of post-translational processing of recombinant products, and efficient purification methods for product recovery. This review discusses recent biotechnology developments for plant-derived monoclonal antibodies and discusses these products as a promising approach to rabies prophylaxis and the consequence for global health benefits. Publication Types: Research Support, U.S. Gov't, Non-P.H.S. Review PMID: 15896408 [PubMed - indexed for MEDLINE] 1018: J Econ Entomol. 2005 Apr;98(2):248-59. Development of genetic sexing strains in Lepidoptera: from traditional to transgenic approaches. Marec F, Neven LG, Robinson AS, Vreysen M, Goldsmith MR, Nagaraju J, Franz G. Insect Pest Control Section, Joint Food and Agriculture Organization/International Atomic Energy Agency, Division of Nuclear Techniques in Food and Agriculture, International Atomic Energy Agency, A-1400 Vienna, Austria. The sterile insect technique (SIT) is currently being used for the control of many agricultural pests, including some lepidopteran species. The SIT relies on the rearing and release of large numbers of genetically sterile insects into a wild population. The holokinetic chromosomes of Lepidoptera respond differently to radiation than do species where there is a localized centromere. This difference has enabled a variation of the SIT to be developed for Lepidoptera where a substerilizing dose of radiation is given to the insects before their release with the result that a certain level of sterility is inherited by the F1 offspring. The development of genetic sexing strains for fruit flies, enabling the release of males only, has resulted in enormous economic benefits in the mass rearing and has increased the efficiency of the field operations severalfold. This article outlines Mendelian approaches that are currently available to separate large numbers of males and females efficiently for different lepidopteran species and describes their difficulties and constraints. Successful transgenesis in several lepidopteran species opens up new possibilities to develop genetic sexing strains. The proposal to develop genetic sexing strains described in this article takes advantage of the fact that in Lepidoptera, the female is the heterogametic sex, with most species having aWZ sex chromosome pair, whereas the males are ZZ. This means that if a conditional lethal gene can be inserted into the W chromosome, then all females should die after the application of the restrictive condition. The assumptions made to accommodate this model are discussed, and the advantages to be gained for control programs are elucidated. Publication Types: Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. PMID: 15889710 [PubMed - indexed for MEDLINE] 1019: Arch Anim Nutr. 2005 Feb;59(1):1-40. Animal nutrition with feeds from genetically modified plants. Flachowsky G, Chesson A, Aulrich K. Institute of Animal Nutrition, Federal Agricultural Research Centre (FAL), Braunschweig, Germany. gerhard.flachowsky@fal.de Plant breeders have made and will continue to make important contributions toward meeting the need for more and better feed and food. The use of new techniques to modify the genetic makeup of plants to improve their properties has led to a new generation of crops, grains and their by-products for feed. The use of ingredients and products from genetically modified plants (GMP) in animal nutrition properly raises many questions and issues, such as the role of a nutritional assessment of the modified feed or feed additive as part of safety assessment, the possible influence of genetically modified (GM) products on animal health and product quality and the persistence of the recombinant DNA and of the 'novel' protein in the digestive tract and tissues of food-producing animals. During the last few years many studies have determined the nutrient value of GM feeds compared to their conventional counterparts and some have additionally followed the fate of DNA and novel protein. The results available to date are reassuring and reveal no significant differences in the safety and nutritional value of feedstuffs containing material derived from the so-called 1st generation of genetically modified plants (those with unchanged gross composition) in comparison with non-GM varieties. In addition, no residues of recombinant DNA or novel proteins have been found in any organ or tissue samples obtained from animals fed with GMP. These results indicate that for compositionally equivalent GMP routine-feeding studies with target species generally add little to nutritional and safety assessment. However, the strategies devised for the nutritional and safety assessment of the 1st generation products will be much more difficult to apply to 2nd generation GMP in which significant changes in constituents have been deliberately introduced (e.g., increased fatty acids or amino acids content or a reduced concentration of undesirable constituents). It is suggested that studies made with animals will play a much more important role in insuring the safety of these 2nd generation constructs. Publication Types: Review PMID: 15889650 [PubMed - indexed for MEDLINE] 1020: J Agric Food Chem. 2005 May 18;53(10):3958-62. Development of a peptide nucleic acid array platform for the detection of genetically modified organisms in food. Germini A, Rossi S, Zanetti A, Corradini R, Fogher C, Marchelli R. Dipartimento di Chimica Organica e Industriale, Università di Parma, Parco area delle Scienze 17/A, 43100 Parma, Italy. Two previously developed platforms, a multiplex polymerase chain reaction (PCR) and a peptide nucleic acid (PNA) array, the former allowing for the simultaneous detection of five transgenes and two endogenous controls in food and feed matrices and the latter for the assessment of the identity of amplified PCR products, were combined in order to develop a PNA array device for the screening of genetically modified organisms (GMOs) in food. PNA probes were opportunely designed, synthesized, and deposited on commercial slides. The length of the probes as well as the distance of the probes from the surface were evaluated and found to be critical points. The most suitable probes were found to be 15-mer PNAs linked to the slide surface by means of two 2-(2-aminoethoxy)ethoxyacetic acids as spacers. The device was tested on a model system constituted by flour samples containing a mixture of standards at known concentrations of transgenic material, in particular Roundup Ready soybean and Bt11, Bt176, Mon810, and GA21 maize: The DNA was amplified using the specific multiplex PCR method and tested on the PNA array. The method proposed was found to be able to correctly identify every GMO present in the tested samples. Publication Types: Research Support, Non-U.S. Gov't PMID: 15884823 [PubMed - indexed for MEDLINE] 1021: J Agric Food Chem. 2005 May 18;53(10):3789-94. Rapid and reliable detection and identification of GM events using multiplex PCR coupled with oligonucleotide microarray. Xu X, Li Y, Zhao H, Wen SY, Wang SQ, Huang J, Huang KL, Luo YB. College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, China. To devise a rapid and reliable method for the detection and identification of genetically modified (GM) events, we developed a multiplex polymerase chain reaction (PCR) coupled with a DNA microarray system simultaneously aiming at many targets in a single reaction. The system included probes for screening gene, species reference gene, specific gene, construct-specific gene, event-specific gene, and internal and negative control genes. 18S rRNA was combined with species reference genes as internal controls to assess the efficiency of all reactions and to eliminate false negatives. Two sets of the multiplex PCR system were used to amplify four and five targets, respectively. Eight different structure genes could be detected and identified simultaneously for Roundup Ready soybean in a single microarray. The microarray specificity was validated by its ability to discriminate two GM maizes Bt176 and Bt11. The advantages of this method are its high specificity and greatly reduced false-positives and -negatives. The multiplex PCR coupled with microarray technology presented here is a rapid and reliable tool for the simultaneous detection of GM organism ingredients. Publication Types: Research Support, Non-U.S. Gov't PMID: 15884798 [PubMed - indexed for MEDLINE] 1022: Sci Am. 2005 May;292(5):25. Defensive eating. Ariza LM. Publication Types: News PMID: 15882013 [PubMed - indexed for MEDLINE] 1023: Kennedy Inst Ethics J. 2005 Mar;15(1):57-76. Germ-line enhancement of humans and non-humans. Loftis JR. Department of Philosophy, St. Lawrence University, Canton, NY, USA. The current difference in attitude toward germ-line enhancement in humans and nonhumans is unjustified. Society should be more cautious in modifying the genes of nonhumans and more bold in thinking about modifying our own genome. I identify four classes of arguments pertaining to germ-line enhancement: safety arguments, justice arguments, trust arguments, and naturalness arguments. The first three types are compelling, but do not distinguish between human and nonhuman cases. The final class of argument would justify a distinction between human and nonhuman germ-line enhancement; however, this type of argument fails and, therefore, the discrepancy in attitude toward human and nonhuman germ-line enhancement is unjustified. PMID: 15881796 [PubMed - indexed for MEDLINE] 1024: Shokuhin Eiseigaku Zasshi. 2005 Feb;46(1):J4-7. Current status of regulation on GM food in Korea. Park SH. Korea Foof and Drug Administration: 5, Nokbun-dong, Eunpyung-gu, Seoul, Korea. PMID: 15881254 [PubMed - indexed for MEDLINE] 1025: Shokuhin Eiseigaku Zasshi. 2005 Feb;46(1):J1-3. [The present condition and a view of the food inspection using the invader method] [Article in Japanese] Hanazawa R, Yamaguchi T. BML Food Science, Inc.: 1491, Matoba, Kawagoe-shi, Saitama 350-1101, Japan. Publication Types: Review PMID: 15881253 [PubMed - indexed for MEDLINE] 1026: Shokuhin Eiseigaku Zasshi. 2005 Feb;46(1):21-7. [Laboratory-performance study of the notified methods to detect genetically modified papaya (55-1)] [Article in Japanese] Kikuchi H, Watanabe T, Kasama K, Wakui C, Matsuki A, Akiyama H, Maitani T. National Institute of Health Sciences: 1-18-1, Kamiyoga, Setagaya-ku, Tokyo 158-8501, Japan. To investigate important factors affecting the reliability of the analytical results, proficiency tests were attempted for the histochemical method (GUS method) and the qualitative PCR method (PCR method) to detect genetically modified papaya (55-1) in the Japanease official method. The test samples were distributed to twenty-three laboratories that participated in the study and were examined according to the protocol. All the data collected from participating laboratories were statistically analyzed. In the PCR method, one negative sample was detected as positive using detection primers in one laboratory, though the sample was negative when checked using confirmation primers. Contamination might have occurred in the step of the preparation of the PCR sample solution using detection primers. In the GUS method, all the test samples were identified as expected. Thus, all the laboratories reported correct results overall. Publication Types: English Abstract PMID: 15881251 [PubMed - indexed for MEDLINE] 1027: Plant Cell. 2005 Jun;17(6):1829-38. Epub 2005 May 6. Dual role for tomato heat shock protein 21: protecting photosystem II from oxidative stress and promoting color changes during fruit maturation. Neta-Sharir I, Isaacson T, Lurie S, Weiss D. Robert H. Smith Institute of Plant Sciences and Genetics in Agriculture, Faculty of Agricultural, Food, and Environmental Quality Sciences, Hebrew University of Jerusalem, Rehovot 76100, Israel. The tomato (Lycopersicon esculentum) chloroplast small heat shock protein (sHSP), HSP21, is induced by heat treatment in leaves, but also under normal growth conditions in developing fruits during the transition of chloroplasts to chromoplasts. We used transgenic tomato plants constitutively expressing HSP21 to study the role of the protein under stress conditions and during fruit maturation. Although we did not find any effect for the transgene on photosystem II (PSII) thermotolerance, our results show that the protein protects PSII from temperature-dependent oxidative stress. In addition, we found direct evidence of the protein's role in fruit reddening and the conversion of chloroplasts to chromoplasts. When plants were grown under normal growth temperature, transgenic fruits accumulated carotenoids earlier than controls. Furthermore, when detached mature green fruits were stored for 2 weeks at 2 degrees C and then transferred to room temperature, the natural accumulation of carotenoids was blocked. In a previous study, we showed that preheat treatment, which induces HSP21, allowed fruit color change at room temperature, after a cold treatment. Here, we show that mature green transgenic fruits constitutively expressing HSP21 do not require the heat treatment to maintain the ability to accumulate carotenoids after cold storage. This study demonstrates that a sHSP plays a role in plant development under normal growth conditions, in addition to its protective effect under stress conditions. Publication Types: Research Support, Non-U.S. Gov't PMID: 15879560 [PubMed - indexed for MEDLINE] 1028: Br J Nutr. 2005 Apr;93 Suppl 1:S91-8. The SYNCAN project: goals, set-up, first results and settings of the human intervention study. Van Loo J, Clune Y, Bennett M, Collins JK. Orafti, Tienen, Belgium. jan.van.loo@orafti.com Experimental evidence on the anticancer properties of dietary prebiotics such as chicory inulin and oligofructose and dietary probiotics has accumulated in recent years. Various experimental models ranging from chemoprevention studies, tumour implantation models to genetically modified mice models, etc. have systematically shown the protective effects of these food ingredients. In some studies it appeared that synbiotics (combination of pre- and probiotics) exerted synergistic activity against processes of carcinogenesis. The logical next step in research was to find out if these observations also would be valid for human volunteers. This was the principal goal of the EU-sponsored SYNCAN project (QLK1-1999-346) which involved the integration of an in vitro study to select the most suitable synbiotic preparation, the application of this synbiotic in an in vivo rat model of chemically induced colon cancer, and, as the heart of the project, the investigation of the synbiotic effects in a human intervention study. The in vitro tests consisted of fermentation studies where the interaction of pre- and probiotics was studied. Cell-free supernatants were generated from various synbiotic combinations fermented by faecal slurry, which were then used to optimise a series of bioassays. In the rat study the anticarcinogenic effect of prebiotics and synbiotics but not of probiotics was demonstrated. Using tissue samples generated in this model, attempts were made to gain a better insight into the mechanisms underlying cancer development. The human intervention study consisted of two groups of volunteers. One group was composed of people at high risk (polypectomised subjects) for colon cancer and the other of volunteers (colon cancer subjects) who had previously undergone 'curative resection' for colon cancer but were not currently receiving treatment. The present paper describes the experimental design of the SYNCAN study, and demonstrates a functional effect of the synbiotic preparation (probiotic survival during gastrointestinal transit and modification of the intestinal flora). Detailed experimental outcome of the human intervention study will be reported elsewhere. Publication Types: Research Support, Non-U.S. Gov't PMID: 15877901 [PubMed - indexed for MEDLINE] 1029: Immunol Cell Biol. 2005 Jun;83(3):271-7. Expression systems and developments in plant-made vaccines. Rigano MM, Walmsley AM. The Biodesign Institute at Arizona State University, School of Life Sciences, Arizona State University, Tempe, 85287, USA. Delivery of vaccines to mucosal surfaces can elicit humoral and cell-mediated responses of the mucosal and systemic immune systems, evoke less pain and discomfort than parenteral delivery, and eliminate needle-associated risks. Transgenic plants are an ideal means by which to produce oral vaccines, as the rigid walls of the plant cell protect antigenic proteins from the acidic environment of the stomach, enabling intact antigen to reach the gut associated lymphoid tissue. In the past few years, new techniques (such as chloroplast transformation and food processing) have improved antigen concentration in transgenic plants. In addition, adjuvants and targeting proteins have increased the immunogenicity of mucosally administered plant-made vaccines. These studies have moved plant-made vaccines closer to the development phase. PMID: 15877605 [PubMed - indexed for MEDLINE] 1030: Risk Anal. 2005 Apr;25(2):467-79. Using surveys in public participation processes for risk decision making: the case of the 2003 British GM Nation? Public debate. Pidgeon NF, Poortinga W, Rowe G, Jones TH, Walls J, O'Riordan T. Centre for Environmental Risk, School of Environmental Sciences, University of East Anglia, Norwich NR4 7TJ, UK. n.pidgeon@uea.ac.uk This article takes as its case study the "GM Nation?" public debate, a major participation process on the commercialization of agricultural biotechnology, which occurred in Britain during the summer of 2003. We investigate possible self-selection biases in over 36,000 open questionnaire responses on the risks and benefits of genetically modified crops and food obtained during GM Nation? A comparison sample of equivalent responses from a statistically representative sample (n = 1,363) of the British general public obtained shortly after the conclusion of the debate is reported. This comparison shows that the GM Nation? open responses were indeed not fully representative of British "public opinion" regarding agricultural biotechnology. Rather, such opinion is not a unitary whole, but fragmented, with considerable ambivalence coexisting alongside outright opposition to GM agriculture. The methodological implications for multistage participation processes are discussed: in particular, the need to anticipate outcomes of complex design decisions, and to include representative public surveys as standard where measures of broader public attitudes to risk are an important objective. Publication Types: Research Support, Non-U.S. Gov't PMID: 15876218 [PubMed - indexed for MEDLINE] 1031: Nature. 2005 May 5;435(7038):3. Pesticide results help China edge transgenic rice towards market. Cyranoski D. Publication Types: News PMID: 15874979 [PubMed - indexed for MEDLINE] 1032: Trends Biotechnol. 2005 May;23(5):222-4. Genetically modified organisms and the EU. Jank B, Rath J, Spök A. Publication Types: Letter PMID: 15865998 [PubMed - indexed for MEDLINE] 1033: Transgenic Res. 2005 Feb;14(1):57-67. Unexpected effects of chitinases on the peach-potato aphid (Myzus persicae Sulzer) when delivered via transgenic potato plants (Solanum tuberosum Linné) and in vitro. Saguez J, Hainez R, Cherqui A, Van Wuytswinkel O, Jeanpierre H, Lebon G, Noiraud N, Beaujean A, Jouanin L, Laberche JC, Vincent C, Giordanengo P. Biologie des Entomophages (UPRES EA 3900), Université de Picardie Jules Verne, 33 rue Saint Leu, 80039 Amiens Cedex, France. With the aim of producing insect-resistant potato plants, internode explants of Solanum tuberosum L. cv. Désirée were transformed with an Agrobacterium strain C58pMP90 containing an insect (Phaedon cochleariae: Coleoptera, Chrysomelidae) chitinase gene and the neomycin phosphotransferase (nptII) gene as selectable marker, both under the control of the viral CaMV 35S promoter. Three transformed potato lines (CH3, CH5 and CH25) exhibiting the highest chitinolytic activities were selected for feeding experiments with the peach-potato aphid, Myzus persicae (Sulzer), under controlled photoperiod and temperature conditions. Aphids fed on transgenic potato plants showed a reduced pre-reproductive period and an enhanced daily fecundity. Transgenic potato lines did not affect nymphal mortality, but improved several biological parameters related to aphid population's growth. Artificial diets were used to provide active (1, 10, 100 and 500 microg ml(-1)) and inactive (500 microg ml(-1)) bacterial (Serratia marcescens) chitinase to M. persicae. These compounds increased nymph survival at all active chitinase doses when compared to the control diet, while inactive chitinase did not. Although the pre-reproductive period was slightly shortened and the daily fecundity slightly higher, active and inactive chitinase provided as food led a reduction from 1 to 1.5 day population's doubling time. Therefore chitinase activity was responsible for the probiotic effects on aphids. Our results question the relevance of a chitinase-based strategy in the context of potato culture protection. Publication Types: Research Support, Non-U.S. Gov't PMID: 15865049 [PubMed - indexed for MEDLINE] 1034: Transgenic Res. 2005 Feb;14(1):15-26. Transformation of fruit trees. Useful breeding tool or continued future prospect? Petri C, Burgos L. Departamento de Mejora y Patología Vegetal, CEBAS-CSIC. Aptd. 164, 30100 Murcia, Spain. Regeneration and transformation systems using mature plant material of woody fruit species have to be achieved as a necessary requirement for the introduction of useful genes into specific cultivars and the rapid evaluation of resulting horticultural traits. Although the commercial production of transgenic annual crops is a reality, commercial genetically-engineered fruit trees are still far from common. In most woody fruit species, transformation and regeneration of commercial cultivars are not routine, generally being limited to a few genotypes or to seedlings. The future of genetic transformation as a tool for the breeding of fruit trees requires the development of genotype-independent procedures, based on the transformation of meristematic cells with high regeneration potential and/or the use of regeneration-promoting genes. The public concern with the introduction of antibiotic resistance into food and the restrictions due to new European laws that do not allow deliberate release of plants transformed with antibiotic-resistance genes highlight the development of methods that avoid the use of antibiotic-dependent selection or allow elimination of marker genesfrom the transformed plant as a research priority in coming years. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 15865045 [PubMed - indexed for MEDLINE] 1035: J AOAC Int. 2005 Mar-Apr;88(2):654-64. Detection of genetically modified corn (Bt176) in spiked cow blood samples by polymerase chain reaction and immunoassay methods. Petit L, Baraige F, Bertheau Y, Brunschwig P, Diolez A, Duhem K, Duplan MN, Fach P, Kobilinsky A, Lamart S, Schattner A, Martin P. Agence Française de Sécurité Sanitaire des Aliments, Unité d'Etude Moléculaire des Contaminants Biologiques Alimentaires (EBA), 94700 Maisons-Alfort, France. l.petit@afssa.fr The fate of DNA and protein transgenic sequences in products derived from animals fed transgenic crops has recently raised public interest. Sensitive molecular tests targeting the Bt176 genetic construct and the transgenic Cry1Ab protein were developed to determine whether plant sequences, especially transgenic sequences, are present in animal products. A protocol for total DNA extraction and purification from cow whole blood samples was first drawn up and assessed by spiking with known amounts of DNA from Bt176 maize. The limit of detection for transgenic sequences (35S promoter and Bt176-specific junction sequence) was determined by both the polymerase chain reaction-enzyme-linked immunosorbent assay (PCR-ELISA) and the 5'-nuclease PCR assay. Four additional PCR systems were built to substantiate the results. The first detects a mono-copy maize-specific sequence (ADH promoter). Two others target multi-copy sequences from plant nucleus (26S rRNA gene) and chloroplast (psaB gene). The last one, used as a positive control, targets a mono-copy animal sequence (alpha(s1)-casein gene). Both methods detected a minimum spiking at 25 copies of Bt176 maize/mL in 10 mL whole blood samples. The sandwich ELISA kit used detected down to 1 ng transgenic Cry1Ab protein/mL spiked whole blood. Publication Types: Comparative Study Research Support, Non-U.S. Gov't PMID: 15861534 [PubMed - indexed for MEDLINE] 1036: Science. 2005 Apr 29;308(5722):688-90. Comment in: Science. 2005 Oct 14;310(5746):231-3; author reply 231-3. Science. 2005 Oct 14;310(5746):231-3; author reply 231-3. Science. 2005 Oct 14;310(5746):231-3; author reply 231-3. Insect-resistant GM rice in farmers' fields: assessing productivity and health effects in China. Huang J, Hu R, Rozelle S, Pray C. Center for Chinese Agricultural Policy, Institute of Geographical Sciences and Natural Resource Research, Chinese Academy of Sciences, Jia 11, Datun Road, Beijing 100101, China. jkhuang.ccap@igsnrr.ac.cn Although no country to date has released a major genetically modified (GM) food grain crop, China is on the threshold of commercializing GM rice. This paper studies two of the four GM varieties that are now in farm-level preproduction trials, the last step before commercialization. Farm surveys of randomly selected farm households that are cultivating the insect-resistant GM rice varieties, without the aid of experimental station technicians, demonstrate that when compared with households cultivating non-GM rice, small and poor farm households benefit from adopting GM rice by both higher crop yields and reduced use of pesticides, which also contribute to improved health. Publication Types: Research Support, Non-U.S. Gov't PMID: 15860626 [PubMed - indexed for MEDLINE] 1037: J AOAC Int. 2005 Mar-Apr;88(2):558-73. Quantitation of 35S promoter in maize DNA extracts from genetically modified organisms using real-time polymerase chain reaction, part 2: interlaboratory study. Feinberg M, Fernandez S, Cassard S, Bertheau Y. Institut National de la Recherche Agronomique, 16 Rue Claude Bernard, 75231 Paris, France. feinberg@inapg.inra.fr The European Committee for Standardization (CEN) and the European Network of GMO Working Laboratories have proposed development of a modular strategy for stepwise validation of complex analytical techniques. When applied to the quantitation of genetically modified organisms (GMOs) in food products, the instrumental quantitation step of the technique is separately validated from the DNA extraction step to better control the sources of uncertainty and facilitate the validation of GMO-specific polymerase chain reaction (PCR) tests. This paper presents the results of an interlaboratory study on the quantitation step of the method standardized by CEN for the detection of a regulatory element commonly inserted in GMO maize-based foods. This is focused on the quantitation of P35S promoter through using the quantitative real-time PCR (QRT-PCR). Fifteen French laboratories participated in the interlaboratory study of the P35S quantitation operating procedure on DNA extract samples using either the thermal cycler ABI Prism 7700 (Applied Biosystems, Foster City, CA) or Light Cycler (Roche Diagnostics, Indianapolis, IN). Attention was focused on DNA extract samples used to calibrate the method and unknown extract samples. Data were processed according to the recommendations of ISO 5725 standard. Performance criteria, obtained using the robust algorithm, were compared to the classic data processing after rejection of outliers by the Cochran and Grubbs tests. Two laboratories were detected as outliers by the Grubbs test. The robust precision criteria gave values between the classical values estimated before and after rejection of the outliers. Using the robust method, the relative expanded uncertainty by the quantitation method is about 20% for a 1% Bt176 content, whereas it can reach 40% for a 0.1% Bt176. The performances of the quantitation assay are relevant to the application of the European regulation, which has an accepted tolerance interval of about +/-50%. These data were fitted to a power model (r2 = 0.96). Thanks to this model, it is possible to propose an estimation of uncertainty of the QRT-PCR quantitation step and an uncertainty budget depending on the analytical conditions. Publication Types: Multicenter Study Research Support, Non-U.S. Gov't PMID: 15859084 [PubMed - indexed for MEDLINE] 1038: J AOAC Int. 2005 Mar-Apr;88(2):536-46. Characterization and event specific-detection by quantitative real-time PCR of T25 maize insert. Collonnier C, Schattner A, Berthier G, Boyer F, Coué-Philippe G, Diolez A, Duplan MN, Fernandez S, Kebdani N, Kobilinsky A, Romaniuk M, de Beuckeleer M, de Loose M, Windels P, Bertheau Y. Laboratoire de Méthodologies de la Détection des OGM, Institut National de la Recherche Agronomique, Route de Saint Cyr, Versailles, France. T25 is one of the 4 maize transformation events from which commercial lines have so far been authorized in Europe. It was created by polyethylene glycol-mediated transformation using a construct bearing one copy of the synthetic pat gene associated with both promoter and terminator of the 35S ribosomal gene from cauliflower mosaic virus. In this article, we report the sequencing of the whole T25 insert and the characterization of its integration site by using a genome walking strategy. Our results confirmed that one intact copy of the initial construct had been integrated in the plant genome. They also revealed, at the 5' junction of the insert, the presence of a second truncated 35S promoter, probably resulting from rearrangements which may have occurred before or during integration of the plasmid DNA. The analysis of the junction fragments showed that the integration site of the insert presented high homologies with the Huck retrotransposon family. By using one primer annealing in the maize genome and the other in the 5' end of the integrated DNA, we developed a reliable event-specific detection system for T25 maize. To provide means to comply with the European regulation, a real-time PCR test was designed for specific quantitation of T25 event by using Taqman chemistry. Publication Types: Research Support, Non-U.S. Gov't PMID: 15859082 [PubMed - indexed for MEDLINE] 1039: Appl Microbiol Biotechnol. 2005 Aug;68(3):292-304. Epub 2005 Apr 26. The use of genetically modified Saccharomyces cerevisiae strains in the wine industry. Schuller D, Casal M. Centro de Biologia (CB-UM), Departamento de Biologia, Universidade do Minho, Braga, Portugal. dschuller@bio.uminho.pt In recent decades, science and food technology have contributed at an accelerated rate to the introduction of new products to satisfy nutritional, socio-economic and quality requirements. With the emergence of modern molecular genetics, the industrial importance of Saccharomyces cerevisiae, is continuously extended. The demand for suitable genetically modified (GM) S. cerevisiae strains for the biofuel, bakery and beverage industries or for the production of biotechnological products (e.g. enzymes, pharmaceutical products) will continuously grow in the future. Numerous specialised S. cerevisiae wine strains were obtained in recent years, possessing a wide range of optimised or novel oenological properties, capable of satisfying the demanding nature of modern winemaking practise. The unlocking of transcriptome, proteome and metabolome complexities will contribute decisively to the knowledge about the genetic make-up of commercial yeast strains and will influence wine strain improvement via genetic engineering. The most relevant advances regarding the importance and implications of the use of GM yeast strains in the wine industry are discussed in this mini-review. In this work, various aspects are considered including the strategies used for the construction of strains with respect to current legislation requirements, the environmental risk evaluations concerning the deliberate release of genetically modified yeast strains, the methods for detection of recombinant DNA and protein that are currently under evaluation, and the reasons behind the critical public perception towards the application of such strains. Publication Types: Review PMID: 15856224 [PubMed - indexed for MEDLINE] 1040: J Agric Food Chem. 2005 May 4;53(9):3333-7. Interlaboratory transfer of a PCR multiplex method for simultaneous detection of four genetically modified maize lines: Bt11, MON810, T25, and GA21. Hernández M, Rodríguez-Lázaro D, Zhang D, Esteve T, Pla M, Prat S. Departament de Genètica Molecular, Institut de Biologia Molecular de Barcelona, Centro de Investigación y Desarrollo-Consejo Superior de Investigaciones Científicas, 08034 Barcelona, Spain. The number of cultured hectares and commercialized genetically modified organisms (GMOs) has increased exponentially in the past 9 years. Governments in many countries have established a policy of labeling all food and feed containing or produced by GMOs. Consequently, versatile, laboratory-transferable GMO detection methods are in increasing demand. Here, we describe a qualitative PCR-based multiplex method for simultaneous detection and identification of four genetically modified maize lines: Bt11, MON810, T25, and GA21. The described system is based on the use of five primers directed to specific sequences in these insertion events. Primers were used in a single optimized multiplex PCR reaction, and sequences of the amplified fragments are reported. The assay allows amplification of the MON810 event from the 35S promoter to the hsp intron yielding a 468 bp amplicon. Amplification of the Bt11 and T25 events from the 35S promoter to the PAT gene yielded two different amplicons of 280 and 177 bp, respectively, whereas amplification of the 5' flanking region of the GA21 gave rise to an amplicon of 72 bp. These fragments are clearly distinguishable in agarose gels and have been reproduced successfully in a different laboratory. Hence, the proposed method comprises a rapid, simple, reliable, and sensitive (down to 0.05%) PCR-based assay, suitable for detection of these four GM maize lines in a single reaction. Publication Types: Research Support, Non-U.S. Gov't PMID: 15853368 [PubMed - indexed for MEDLINE] 1041: Genewatch. 2005 Jan-Feb;18(1):12-4, 18. The genetic bill of rights: advancing a rights platform in biotechnology. Krimksy S, Shorett P. Council for Responsible Genetics (CRG) Board, USA. PMID: 15838997 [PubMed - indexed for MEDLINE] 1042: Nature. 2005 Apr 14;434(7035):807. Comment in: Nature. 2005 Jun 2;435(7042):561. Don't rely on Uncle Sam. [No authors listed] Publication Types: Editorial PMID: 15829921 [PubMed - indexed for MEDLINE] 1043: J Anim Sci. 2005 May;83(5):1068-74. Glufosinate herbicide-tolerant (LibertyLink) rice vs. conventional rice in diets for growing-finishing swine. Cromwell GL, Henry BJ, Scott AL, Gerngross MF, Dusek DL, Fletcher DW. University of Kentucky, Lexington, 40546, USA. gcromwel@uky.edu Genetically modified (GM) rice (LibertyLink, event LLRICE62) that is tolerant to glufosinate ammonium (Liberty) herbicide was compared with a near-isogenic (NI) conventional medium-grain brown rice (cultivar, Bengal) and a commercially milled long-grain brown rice in diets for growing-finishing pigs. The GM and NI rice were grown in 2000. The GM rice was from fields treated (GM+) or not treated (GM-) with glufosinate herbicide. The GM- and NI rice were grown using herbicide regimens typical of southern United States rice production practices. The four rice grains were similar in composition. Growing-finishing pigs (n = 96) were fed fortified rice-soybean meal diets containing the four different rice grains from 25 to 106 kg BW. Diets contained 0.99% lysine initially (growing phase), with lysine decreased to 0.80% (early finishing phase) and 0.65% (late finishing phase), when pigs reached 51 and 77 kg, respectively. The percentage of rice in the four diets was constant during each of the three phases (72.8, 80.0, and 85.8% for the growing, early-finishing, and late-finishing phases, respectively). There were six pen replicates (three pens of barrows and three pens of gilts) and four pigs per pen for each dietary treatment. All pigs were slaughtered at the termination of the study to collect carcass data. At the end of the 98-d experiment, BW gain, feed intake (as-fed basis), and feed:gain ratio did not differ (P > 0.05) for pigs fed the GM+ vs. conventional rice diets, but growth performance traits of pigs fed the GM+ rice diets were superior (P < 0.05) to those of pigs fed the GM- rice diet (ADG = 0.86, 0.79, 0.81, and 0.85 kg/d; ADFI = 2.41, 2.49, 2.37, and 2.45 kg/d; feed:gain = 2.80, 3.17, 2.95, and 2.89 for GM+, GM-, NI, and commercially milled rice, respectively). Carcass traits (adjusted for final BW) did not differ (P = 0.10) among treatments (hot carcass yield = 73.5, 72.6, 72.6, and 73.2%; 10th-rib backfat = 23.0, 22.7, 21.3, and 23.8 mm; LM area = 38.6, 38.0, 38.2, and 38.1 cm(2); carcass fat-free lean = 50.5, 50.5, 51.2, and 50.0%). Gilts grew slower (P < 0.05) and were leaner (P < 0.05) than barrows. Responses to type of rice did not differ between barrows and gilts, with no evidence of a diet x gender interaction (P = 0.50) for any trait. The results indicate that the glufosinate herbicide-tolerant rice was similar in composition and nutritional value to conventional rice for growing-finishing pigs. Publication Types: Comparative Study PMID: 15827252 [PubMed - indexed for MEDLINE] 1044: J Agric Food Chem. 2005 Apr 20;53(8):3041-52. Event-specific plasmid standards and real-time PCR methods for transgenic Bt11, Bt176, and GA21 maize and transgenic GT73 canola. Taverniers I, Windels P, Vaïtilingom M, Milcamps A, Van Bockstaele E, Van den Eede G, De Loose M. Department for Plant Genetics and Breeding, Centre for Agricultural Research, Caritasstraat 21, B-9090 Melle, Belgium. i.taverniers@clo.fgov.be Since the 18th of April 2004, two new regulations, EC/1829/2003 on genetically modified food and feed products and EC/1830/2003 on traceability and labeling of GMOs, are in force in the EU. This new, comprehensive regulatory framework emphasizes the need of an adequate tracing system. Unique identifiers, such as the transgene genome junction region or a specific rearrangement within the transgene DNA, should form the basis of such a tracing system. In this study, we describe the development of event-specific tracing systems for transgenic maize lines Bt11, Bt176, and GA21 and for canola event GT73. Molecular characterization of the transgene loci enabled us to clone an event-specific sequence into a plasmid vector, to be used as a marker, and to develop line-specific primers. Primer specificity was tested through qualitative PCRs and dissociation curve analysis in SYBR Green I real-time PCRs. The primers were then combined with event-specific TaqMan probes in quantitative real-time PCRs. Calibration curves were set up both with genomic DNA samples and the newly synthesized plasmid DNA markers. It is shown that cloned plasmid GMO target sequences are perfectly suitable as unique identifiers and quantitative calibrators. Together with an event-specific primer pair and a highly specific TaqMan probe, the plasmid markers form crucial components of a unique and straighforward tracing system for Bt11, Bt176, and GA21 maize and GT73 canola events. Publication Types: Research Support, Non-U.S. Gov't PMID: 15826057 [PubMed - indexed for MEDLINE] 1045: Nat Biotechnol. 2005 Apr;23(4):429-30. Comment in: Nat Biotechnol. 2006 Oct;24(10):1200-1. Comment on: Nat Biotechnol. 2005 Apr;23(4):482-7. Golden Rice gets a boost from maize. Grusak MA. USDA-ARS Children's Nutrition Research Center, Department of Pediatrics, Baylor College of Medicine, 1100 Bates Street, Houston, TX 77030-2600, USA. mgrusak@bcm.tmc.edu Publication Types: Comment Research Support, U.S. Gov't, Non-P.H.S. PMID: 15815666 [PubMed - indexed for MEDLINE] 1046: Nat Biotechnol. 2005 Apr;23(4):403. Monsanto branches out into fruits and vegetables. Herrera S. Publication Types: News PMID: 15815653 [PubMed - indexed for MEDLINE] 1047: Allergy. 2005 May;60(5):559-64. Risks of allergic reactions to biotech proteins in foods: perception and reality. Lehrer SB, Bannon GA. Section of Clinical Immunology, Allergy and Rheumatology, Tulane University School of Medicine, New Orleans, LA 70112, USA. In recent years, significant attention has been paid to the use of biotechnology to improve the quality and quantity of the food supply due in part to the projected growth in the world population, plus limited options available for increasing the amount of land under cultivation. Alterations in the food supply induced by classical breeding and selection methods typically involve the movement of large portions of genomic DNA between different plant varieties to obtain the desired trait. This is in contrast to techniques of genetic engineering which allows the selection and transfers specific genes from one species to another. The primary allergy risk to consumers from genetically modified crops may be placed into one of three categories. The first represents the highest risk to the allergic consumer is the transfer of known allergen or cross-reacting allergen into a food crop. The second category, representing an intermediate risk to the consumer, is the potential for replacing the endogenous allergenicity of a genetically-modified crop. The last category involves expression of novel proteins that may become allergens in man and generally represents a relatively low risk to the consumer, although this possibility has received attention of late. In order to mitigate the three categories of potential allergy risk associated with biotech crops, all genes introduced into food crops undergo a series of tests designed to determine if the biotech protein exhibits properties of known food allergens. The result of this risk assessment process to date is that no biotech proteins in foods have been documented to cause allergic reactions. These results indicate that the current assessment process is robust, although as science of allergy and allergens evolves, new information and new technology should help further the assessment process for potential allergenicity. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 15813800 [PubMed - indexed for MEDLINE] 1048: Protein Expr Purif. 2005 May;41(1):53-60. Refolding and purification of non-fusion HPT protein expressed in Escherichia coli as inclusion bodies. Zhuo Q, Piao JH, Wang R, Yang XG. Institute of Nutrition and Food Safety, Chinese Center for Disease Control and Prevention, Beijing 100050, China. zhuoqin1@sina.com.cn The gene encoding hygromycin B phosphotransferase (hpt) is a widely used selectable marker in the production of genetically engineered crops. To facilitate the safety assessment of this protein, the non-fusion hpt expression plasmid was constructed and introduced into Escherichia coli to produce enough quantity of the HPT protein. High level expressed HPT was achieved but most of the expressed protein aggregated as inclusion bodies. The inclusion bodies were washed, separated from the cells, and solubilized by 0.3% Sarkosyl. The protein was renatured by dilution and dialysis, and then purified by anion-exchange chromatography. The activity is 8 U/mg protein and the purity is about 95%. Further studies showed that the microbially produced HPT protein had comparable molecular weight, immuno-reactivities, N-terminal amino acid sequences, and biological activities with those of the HPT produced by transgenic rice harboring hpt gene. All these results demonstrated the validity of utilizing the microbially produced HPT to assess the safety of the HPT protein produced in genetically engineered rice. Publication Types: Research Support, Non-U.S. Gov't PMID: 15802221 [PubMed - indexed for MEDLINE] 1049: Nature. 2005 Mar 31;434(7033):548. Stray seeds had antibiotic-resistance genes. Macilwain C. Publication Types: News PMID: 15800583 [PubMed - indexed for MEDLINE] 1050: Rejuvenation Res. 2005 Spring;8(1):37-45. Genetically modified hairy roots of Withania somnifera Dunal: a potent source of rejuvenating principles. Kumar V, Murthy KN, Bhamid S, Sudha CG, Ravishankar GA. Plant Cell Biotechnology Department, Central Food Technological Research Institute, Mysore, Karnataka, India. Transgenic hairy roots were induced from Withania somnifera Dunal, by infecting leaf explants with Agrobacterium rhizogenes. Polymerase chain reaction for rol A gene and Southern blot confirmed the integration of T-DNA in the genome. Cultures were grown in Murashige and Skoog solid as well as in liquid medium. The antioxidant activity was assayed in roots grown in solid media and liquid media. Hairy roots grown in liquid media found to possess highly significant activity in 1,1-diphenyl-2-pecryl-hydrazyl radical, beta-carotene linoleic acid model system. The activity was 57.34%, 75.64%, and 93.41% in case DPPH model and 55.3%, 76.3%, and 90.5% in case of b-CLAMS in 25, 50, and 100 mg L(-1) concentration, respectively. In case of hydroxyl radical trapping and brain lipid peroxidation assay, the activity was more significant in hairy roots grown on solid medium in comparison with commercial formulation prepared using normal roots and standard withanaloids. Root extract grown in solid medium has shown 93.2% hydroxyl radical trapping activity at 100 mg L(-1) concentration, and 500 mg L(-1) has shown 83.6% in case of brain lipid peroxidation assay. High-performance liquid chromatography analysis demonstrated the presence of withanaloids in the hairy root extracts. The results of the study clearly indicate that there is enhancement of secondary metabolites in hairy roots, which is indicated through significant enhancement of the antioxidant activity, since these are the major constituents responsible for the activity. This is the first report on the presence of antioxidant principles in genetically modified roots of W. somnifera. These results of the present study may aid in utilization of the W. somnifera hairy roots for its rejuvenating principles. Publication Types: Research Support, Non-U.S. Gov't PMID: 15798373 [PubMed - indexed for MEDLINE] 1051: Nature. 2005 Mar 24;434(7032):423. US launches probe into sales of unapproved transgenic corn. Macilwain C. Publication Types: News PMID: 15791213 [PubMed - indexed for MEDLINE] 1052: Plant Cell Rep. 2005 Jun;24(4):209-15. Epub 2005 Mar 24. Agrobacterium tumefaciens-mediated transformation of leek (Allium porrum) and garlic (Allium sativum). Eady C, Davis S, Catanach A, Kenel F, Hunger S. New Zealand Institute for Crop & Food Research Limited, Private bag Box 4704, Christchurch, New Zealand. eadyc@crop.cri.nz Transgenic leek (Allium porrum) and garlic (Allium sativum) plants have been recovered by the selective culturing of immature leek and garlic embryos via Agrobacterium-mediated transformation using a method similar to that described by Eady et al. (Plant Cell Rep 19:376-381, 2000) for onion transformation. This method involved the use of a binary vector containing the m-gfp-ER reporter gene and nptII selectable marker, and followed the protocol developed previously for the transformation of onions with only minor modifications pertaining to the post-transformation selection procedure which was simplified to have just a single selection regime. Transgenic cultures were selected for their ability to express the m-gfp-ER reporter gene and grown in the presence of geneticin (20 mg/l). The presence of transgenes in the genome of the plants was confirmed using TAIL-PCR and Southern analysis. This is the first report of leek and "true seed" garlic transformation. It now makes possible the integration of useful agronomic and quality traits into these crops. PMID: 15789208 [PubMed - indexed for MEDLINE] 1053: Risk Anal. 2005 Feb;25(1):199-209. Trust in risk regulation: cause or consequence of the acceptability of GM food? Poortinga W, Pidgeon NF. Centre for Environmental Risk, University of East Anglia, Norwich NR4 7TJ, UK. w.poortinga@uea.ac.uk Although there is ample empirical evidence that trust in risk regulation is strongly related to the perception and acceptability of risk, it is less clear what the direction of this relationship is. This article explores the nature of the relationship, using three separate data sets on perceptions of genetically modified (GM) food among the British public. The article has two discrete but closely interrelated objectives. First, it compares two models of trust. More specifically, it investigates whether trust is the cause (causal chain account) or the consequence (associationist view) of the acceptability of GM food. Second, this study explores whether the affect heuristic can be applied to a wider number of risk-relevant concepts than just perceived risk and benefit. The results suggest that, rather than a determinant, trust is an expression or indicator of the acceptability of GM food. In addition, and as predicted, "affect" accounts for a large portion of the variance between perceived risk, perceived benefit, trust in risk regulation, and acceptability. Overall, the results support the associationist view that specific risk judgments are driven by more general evaluative judgments The implications of these results for risk communication and policy are discussed. Publication Types: Research Support, Non-U.S. Gov't PMID: 15787769 [PubMed - indexed for MEDLINE] 1054: J Chromatogr A. 2005 Feb 11;1065(1):107-13. Detection of processed genetically modified food using CIM monolithic columns for DNA isolation. Jerman S, Podgornik A, Cankar K, Cadet N, Skrt M, Zel J, Raspor P. Department of Food Science and Technology, Biotechnical Faculty, University of Ljubljana, Jamnikarjeva 101, S-1000 Ljubljana, Slovenia. The availability of sufficient quantities of DNA of adequate quality is crucial in polymerase chain reaction (PCR)-based methods for genetically modified food detection. In this work, the suitability of anion-exchange CIM (Convective Interaction Media; BIA Separations, Ljubljana, Slovenia) monolithic columns for isolation of DNA from food was studied. Maize and its derivates corn meal and thermally pretreated corn meal were chosen as model food. Two commercially available CIM disk columns were tested: DEAE (diethylaminoethyl) and QA (quaternary amine). Preliminary separations were performed with standard solution of salmon DNA at different pH values and different NaCl concentrations in mobile phase. DEAE groups and pH 8 were chosen for further isolations of DNA from a complex matrix-food extract. The quality and quantity of isolated DNA were tested on agarose gel electrophoresis, with UV-scanning spectrophotometry, and by amplification with real-time PCR. DNA isolated in this way was of suitable quality for further PCR analyses. The described method is also applicable for DNA isolation from processed foods with decreased DNA content. Furthermore, it is more effective and less time-consuming in comparison with the existing proposed methods for isolation of DNA from plant-derived foods. Publication Types: Research Support, Non-U.S. Gov't PMID: 15782956 [PubMed - indexed for MEDLINE] 1055: J Agric Food Chem. 2005 Mar 23;53(6):2060-9. Comparative studies of the quantification of genetically modified organisms in foods processed from maize and soy using trial producing. Yoshimura T, Kuribara H, Kodama T, Yamata S, Futo S, Watanabe S, Aoki N, Iizuka T, Akiyama H, Maitani T, Naito S, Hino A. Analytical Technology Laboratory, Asahi Breweries, Ltd., 1-1-21 Midori, Moriya, Ibaraki 302-0106, Japan. Seven types of processed foods, namely, cornstarch, cornmeal, corn puffs, corn chips, tofu, soy milk, and boiled beans, were trial produced from 1 and 5% (w/w) genetically modified (GM) mixed raw materials. In this report, insect resistant maize (MON810) and herbicide tolerant soy (Roundup Ready soy, 40-3-2) were used as representatives of GM maize and soy, respectively. Deoxyribonucleic acid (DNA) was extracted from the raw materials and the trial-produced processed food using two types of methods, i.e., the silica membrane method and the anion exchange method. The GM% values of these samples were quantified, and the significant differences between the raw materials and the trial-produced processed foods were statistically confirmed. There were some significant differences in the comparisons of all processed foods. However, our quantitative methods could be applied as a screening assay to tofu and soy milk because the differences in GM% between the trial-produced processed foods and their raw materials were lower than 13 and 23%, respectively. In addition, when quantitating with two primer pairs (SSIIb 3, 114 bp; SSIIb 4, 83 bp for maize and Le1n02, 118 bp; Le1n03, 89 bp for soy), which were targeted within the same taxon specific DNA sequence with different amplicon sizes, the ratios of the copy numbers of the two primer pairs (SSIIb 3/4 and Le1n02/03) decreased with time in a heat-treated processing model using an autoclave. In this report, we suggest that the degradation level of DNA in processed foods could be estimated from these ratios, and the probability of GM quantification could be experimentally predicted from the results of the trial producing. Publication Types: Comparative Study Research Support, Non-U.S. Gov't PMID: 15769136 [PubMed - indexed for MEDLINE] 1056: J Agric Food Chem. 2005 Mar 23;53(6):2052-9. Applicability of the quantification of genetically modified organisms to foods processed from maize and soy. Yoshimura T, Kuribara H, Matsuoka T, Kodama T, Iida M, Watanabe T, Akiyama H, Maitani T, Furui S, Hino A. Analytical Technology Laboratory, Asahi Breweries, Ltd., 1-1-21 Midori, Moriya, Ibaraki 302-0106, Japan. The applicability of quantifying genetically modified (GM) maize and soy to processed foods was investigated using heat treatment processing models. The detection methods were based on real-time quantitative polymerase chain reaction (PCR) analysis. Ground seeds of insect resistant GM maize (MON810) and glyphosate tolerant Roundup Ready (RR) soy were dissolved in water and were heat treated by autoclaving for various time intervals. The calculated copy numbers of the recombinant and taxon specific deoxyribonucleic acid (DNA) sequences in the extracted DNA solution were found to decrease with time. This decrease was influenced by the PCR-amplified size. The conversion factor (Cf), which is the ratio of the recombinant DNA sequence to the taxon specific DNA sequence and is used as a constant number for calculating GM% at each event, tended to be stable when the sizes of PCR products of two DNA sequences were nearly equal. The results suggested that the size of the PCR product plays a key role in the quantification of GM organisms in processed foods. It is believed that the Cf of the endosperm (3n) is influenced by whether the GM originated from a paternal or maternal source. The embryos and endosperms were separated from the F1 generation seeds of five GM maize events, and their Cf values were measured. Both paternal and maternal GM events were identified. In these, the endosperm Cf was lower than that of the embryo, and the embryo Cf was lower than that of the endosperm. These results demonstrate the difficulties encountered in the determination of GM% in maize grains (F2 generation) and in processed foods from maize and soy. Publication Types: Research Support, Non-U.S. Gov't PMID: 15769135 [PubMed - indexed for MEDLINE] 1057: J Agric Food Chem. 2005 Mar 23;53(6):2028-31. Screening of a peanut (Arachis hypogaea L.) cDNA library to isolate a Bowman-Birk trypsin inhibitor clone. Boateng JA, Viquez OM, Konan KN, Dodo HW. Department of Food and Animal Sciences, Food Biotechnology Laboratory, Alabama A&M University, Normal, Alabama 35762, USA. Peanut crop losses due to insect and pest infestation cost peanut farmers nearly 20% of their annual yields. The conventional use of chemicals to combat this problem is costly and toxic to humans and livestock and leads to the development of resistance by target insects. Transgenic plants expressing a trypsin inhibitor gene in tobacco and cowpea have proven to be efficient for resistance against insects. Therefore, a transgenic peanut overexpressing a trypsin inhibitor gene could be an alternative solution to the use of toxic chemicals. Five Bowman-Birk trypsin inhibitor (BBTI) proteins were previously isolated from peanut. However, to date, neither cDNA nor genomic DNA sequences are available. The objective of this research was to screen a peanut cDNA library to isolate and sequence at least one full-length peanut BBTI cDNA clone. Two heterologous oligonucleotides were constructed on the basis of a garden pea (Pisum sativa) trypsin inhibitor nucleotide sequence and used as probes to screen a peanut lambda gt-11 cDNA library. Two positive and identical cDNA clones were isolated, subcloned into a pBluescript vector, and sequenced. Sequence analysis revealed a full-length BBTI cDNA of about 243 bp, with a start codon ATG at position +1 and a stop codon TGA at position +243. In the 3' end, two poly adenylation signals (AATAAA) were identified at positions +261 and +269. The isolated cDNA clone encodes a protein of 80 amino acid residues including a leader sequence of 11 amino acids. The deduced amino acid sequence is 100% identical to published sequences of peanut BBTI AI, AII, BI, and BIII and 81% identical to BII. PMID: 15769131 [PubMed - indexed for MEDLINE] 1058: Environ Sci Pollut Res Int. 2005;12(1):56. Understanding Western Australian consumers' views: acceptance of food produced using gene technology. A case of herbicide tolerant canola (Brassica napus L.). Baumann A, Osman M, Burton M, Lumley S. School of Agricultural and Resource Economics, University of Western Australia, Nedlands, Western Australia 6009, Australia. eilif@gmx.net PMID: 15768741 [PubMed - indexed for MEDLINE] 1059: Nat Biotechnol. 2005 Mar;23(3):283-5. Erratum in: Nat Biotechnol. 2005 Apr;23(4):488. Clone on the range: What animal biotech is bringing to the table. Dove AW. Publication Types: News PMID: 15765075 [PubMed - indexed for MEDLINE] 1060: Nat Biotechnol. 2005 Mar;23(3):281. Agbio keeps on growing. Lawrence S. PMID: 15765074 [PubMed - indexed for MEDLINE] 1061: J AOAC Int. 2005 Jan-Feb;88(1):136-55. Polymerase chain reaction technology as analytical tool in agricultural biotechnology. Lipp M, Shillito R, Giroux R, Spiegelhalter F, Charlton S, Pinero D, Song P. Monsanto Co., 800 N. Lindbergh Blvd, St. Louis, MO 63167, USA. markus.lipp@monsanto.com The agricultural biotechnology industry applies polymerase chain reaction (PCR) technology at numerous points in product development. Commodity and food companies as well as third-party diagnostic testing companies also rely on PCR technology for a number of purposes. The primary use of the technology is to verify the presence or absence of genetically modified (GM) material in a product or to quantify the amount of GM material present in a product. This article describes the fundamental elements of PCR analysis and its application to the testing of grains. The document highlights the many areas to which attention must be paid in order to produce reliable test results. These include sample preparation, method validation, choice of appropriate reference materials, and biological and instrumental sources of error. The article also discusses issues related to the analysis of different matrixes and the effect they may have on the accuracy of the PCR analytical results. Publication Types: Research Support, Non-U.S. Gov't PMID: 15759736 [PubMed - indexed for MEDLINE] 1062: Commun Agric Appl Biol Sci. 2004;69(3):185-9. Transgenic-Bt potato plant resistance to the colorado potato beetle affect the aphid parasitoid Aphidius nigripes. Ashouri A. Plant Protection Department, Faculty of Agriculture, University of Tehran, CP 31587-11167, Karaj, Iran. ashouri@ut.ac.ir Using the biotechnological plant resistance for herbivore control with less reliance on chemicals in integrated pest management (IPM) programs critically depends on predictable interactions with no-target organisms of various trophic levels. Plant resistance to insect pests based on recombinant Bacillus thuringiensis could interfere with natural enemies of non target pests. Performance of the potato aphid parasitoid Aphidius nigripes was studied on the 'Superior-BT line transgenic for the CryllIA toxin of B. thuringiensis, resistance to the Colorado potato beetle; and none transformed 'Superior' line which served as control. Parasitoid survival was significantly lower on the 'Superior-BT' line compared to control. Adult females were largest on 'Superior' and smallest on BT potatoes. This difference was reflected on parasitoid fecundity, which was lowest on 'Superior-BT', and highest on Superior. The results indicate that factor of potato resistance to the Colorado potato beetle affected the fitness of a parasitold of the aphid Macrosiphum euphorbiae, a secondary pest of potato. The effects on the parasitoid were complex but were generally interpretable in terms of host aphid quality variation among potato lines used as food by the aphids during parasitoid development. Publication Types: Research Support, Non-U.S. Gov't PMID: 15759411 [PubMed - indexed for MEDLINE] 1063: Riv Biol. 2004 Aug-Dec;97(3):379-408. Functional dynamics of living systems and genetic engineering. Buiatti M. Department of Animal Biology and Genetics, Firenze (Italy). The discussion on Genetically Modified Organisms (GMO's) has been centred mainly on the nature and effects on economy, human health, environment, of the few transgenic plant lines present in the market in the last eight years. On the contrary, the present paper starts with a discussion of some of the relevant changes in our basic knowledge of the structure and dynamics of living systems in the last twenty years. Contemporary Biology is then compared with what may be called the "modern paradigm" of life sciences on which present day GMO's are conceptually based. Technical, environmental, social and economic problems deriving from the unexpected, persistent prevalence of the old fashioned modern vision of life in the "spirit of time" will be thoroughly discussed with a particular attention to the virtualisation process of GMO's and the effects of the prevalence over economic, social, environmental reality of their symbolic values. Publication Types: Review PMID: 15754592 [PubMed - indexed for MEDLINE] 1064: Cytogenet Genome Res. 2005;109(1-3):350-9. Detection of alien chromatin introgression from Thinopyrum into wheat using S genomic DNA as a probe--a landmark approach for Thinopyrum genome research. Chen Q. Agriculture and Agri-Food Canada, Lethbridge Research Centre, Lethbridge, Alta, Canada. chenqi@agr.gc.ca The introduction of alien genetic variation from the genus Thinopyrum through chromosome engineering into wheat is a valuable and proven technique for wheat improvement. A number of economically important traits have been transferred into wheat as single genes, chromosome arms or entire chromosomes. Successful transfers can be greatly assisted by the precise identification of alien chromatin in the recipient progenies. Chromosome identification and characterization are useful for genetic manipulation and transfer in wheat breeding following chromosome engineering. Genomic in situ hybridization (GISH) using an S genomic DNA probe from the diploid species Pseudoroegneria has proven to be a powerful diagnostic cytogenetic tool for monitoring the transfer of many promising agronomic traits from Thinopyrum. This specific S genomic probe not only allows the direct determination of the chromosome composition in wheat-Thinopyrum hybrids, but also can separate the Th. intermedium chromosomes into the J, J(S) and S genomes. The J(S) genome, which consists of a modified J genome chromosome distinguished by S genomic sequences of Pseudoroegneria near the centromere and telomere, carries many disease and mite resistance genes. Utilization of this S genomic probe leads to a better understanding of genomic affinities between Thinopyrum and wheat, and provides a molecular cytogenetic marker for monitoring the transfer of alien Thinopyrum agronomic traits into wheat recipient lines. Copyright 2005 S. Karger AG, Basel. Publication Types: Review PMID: 15753596 [PubMed - indexed for MEDLINE] 1065: Trends Plant Sci. 2005 Mar;10(3):112-6. Relief for fish stocks: oceanic fatty acids in transgenic oilseeds. Domergue F, Abbadi A, Heinz E. University of Hamburg, Biozentrum Klein Flottbek, Ohnhorststr. 18, 22609 Hamburg, Germany. fredomergue@voila.fr Three recent reports (Baoxiu Qi et al., Amine Abbadi et al. and Anthony J. Kinney et al.) describe the production of very long-chain polyunsaturated fatty acids in transgenic plants. This might lead to a sustainable source of these valuable fatty acids for use in human food and animal feed. At present they are mainly available via consumption of fish, which is a limited and endangered resource. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 15749468 [PubMed - indexed for MEDLINE] 1066: Regul Toxicol Pharmacol. 2005 Apr;41(3):175-84. Epub 2005 Jan 20. Quantitative measurement of protein digestion in simulated gastric fluid. Herman RA, Korjagin VA, Schafer BW. Dow AgroSciences LLC., 9330 Zionsville Road, Indianapolis, IN 46268, USA. raherman@dow.com The digestibility of novel proteins in simulated gastric fluid is considered to be an indicator of reduced risk of allergenic potential in food, and estimates of digestibility for transgenic proteins expressed in crops are required for making a human-health risk assessment by regulatory authorities. The estimation of first-order rate constants for digestion under conditions of low substrate concentration was explored for two protein substrates (azocoll and DQ-ovalbumin). Data conformed to first-order kinetics, and half-lives were relatively insensitive to significant variations in both substrate and pepsin concentration when high purity pepsin preparations were used. Estimation of digestion efficiency using densitometric measurements of relative protein concentration based on SDS-PAGE corroborated digestion estimates based on measurements of dye or fluorescence release from the labeled substrates. The suitability of first-order rate constants for estimating the efficiency of the pepsin digestion of novel proteins is discussed. Results further support a kinetic approach as appropriate for comparing the digestibility of proteins in simulated gastric fluid. PMID: 15748795 [PubMed - indexed for MEDLINE] 1067: Plant J. 2005 Mar;41(6):791-800. The stability of the Arabidopsis transcriptome in transgenic plants expressing the marker genes nptII and uidA. El Ouakfaoui S, Miki B. Bioproducts and Bioprocesses, Research Branch, Agriculture and Agri-Food Canada, Ottawa, ON K1A 0C6, Canada. The ATH1 Arabidopsis GeneChip from Affymetrix was used to search for transcriptome changes in Arabidopsis associated with the strong expression of transgenes regulated by constitutive promoters. The insertion and expression of the commonly used marker genes, uidA and nptII, did not induce changes to the expression patterns of the approximately 24 000 genes that were screened under optimal growth conditions and under physiological stress imposed by low temperatures. Approximately 8000 genes (35% of the Arabidopsis genome) underwent changes in gene expression in both wild-type and transgenic plants under abiotic stresses such as salt, dehydration, cold, and heat. This study provides detailed information on the extent of non-targeted or pleiotropic effects of transgenes on plants and shows that the transgenic and non-transgenic plants were equivalent in their global patterns of transcription. This information may help to extend our understanding and interpretation of the principle of substantial equivalence which is used as a first step in the biosafety evaluation of transgenic crops. Publication Types: Research Support, Non-U.S. Gov't PMID: 15743445 [PubMed - indexed for MEDLINE] 1068: J Dairy Sci. 2005 Mar;88(3):843-56. Invited review: methods for the screening, isolation, and characterization of exopolysaccharides produced by lactic acid bacteria. Ruas-Madiedo P, de los Reyes-Gavilán CG. Instituto de Productos Lácteos de Asturias, CSIC, Carretera de Infiesto s/n, 33300 Villaviciosa, Asturias, Spain. ruas-madiedo@ipla.csic.es The ability to produce exopolysaccharides (EPS) is widespread among lactic acid bacteria (LAB), although the physiological role of these molecules has not been clearly established yet. Some EPS confer on LAB a "ropy" character that can be detected in cultures that form long strands when extended with an inoculation loop. When EPS are produced in situ during milk fermentation they can act as natural biothickeners, giving the product a suitable consistency, improving viscosity, and reducing syneresis. In addition, some of these EPS may have beneficial effects on human health. The increasing demand by consumers of novel dairy products requires a better understanding of the effect of EPS on existing products and, at the same time, the search for new EPS-producing strains with desirable properties. The use of genetically modified organisms capable of producing high levels of EPS or newly designed biopolymers is still very limited. Therefore, exploration of the biodiversity of wild LAB strains from natural ecological environments is currently the most suitable approach to search for the desired EPS-phenotype. The screening of ropy strains and the isolation and characterization of EPS responsible for this characteristic have led to the application over the past years of a wide variety of techniques. This review summarizes the available information on methods and procedures used for research on this topic. The information provided deals with methods for screening of EPS-producing LAB, detection of the ropy phenotype, and the physicochemical and structural characterization of these molecules, including parameters related to their viscosifying properties. To our knowledge, this is the first compilation of methods available for the study of EPS produced by LAB. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 15738217 [PubMed - indexed for MEDLINE] 1069: Vaccine. 2005 Mar 7;23(15):1866-9. Plant-derived vaccines against diarrheal diseases. Tacket CO. Center for Vaccine Development, University of Maryland School of Medicine, 685 West Baltimore St., Baltimore, MD 21201, USA. ctacket@medicine.umaryland.edu Transgenic plants present a novel system for both production and oral delivery of vaccine antigens. Production of protein antigen in food plants is substantially cheaper than production in bacterial, fungal, insect cell, or mammalian cell culture. Edible plants themselves can also serve as the oral vaccine delivery system. Phase-1 studies of raw transgenic potatoes expressing the B subunit of Escherichia coli heat labile enterotoxin (LT-B), potatoes expressing Norwalk virus capsid protein, and defatted corn germ meal expressing LT-B have been conducted. New oral vaccines based on other transgenic plants will soon be evaluated in humans. Publication Types: Research Support, U.S. Gov't, P.H.S. Review PMID: 15734057 [PubMed - indexed for MEDLINE] 1070: Wei Sheng Yan Jiu. 2004 Nov;33(6):713-5. [Determination of anti-nutrients in genetically modified crops] [Article in Chinese] Han J, Yang Y, Wang Z, Zhou R, Yang X. Institute of Nutrition and Food Safety, Chinese Center for Disease Control and Prevention, Beijing 100050, China. OBJECTIVE: In order to assess the safety of genetically modified crops, the anti-nutrients were determined in these novel crops and compared with their parental lines. METHODS: Five kinds of crops (rice, maize, soybean, cottonseed and rapeseed) from domestic and foreign companies were collected and the contents of anti-nutrients for each kind were analyzed. These anti-nutrients include phytate and protease-inhibitors (in rice, maize and soybean), gossypol (in cottonseed), glucosinolates and erucic acids (in rapeseed). RESULTS AND CONCLUSION: The contents of anti-nutrients in one type of maize, three types of rapeseeds and two types of cottonseeds didn't meet the requirement of "equivalence" in this study, but the finally conclusion should be made after many experimental data in the future. Publication Types: English Abstract Research Support, Non-U.S. Gov't PMID: 15727185 [PubMed - indexed for MEDLINE] 1071: Wei Sheng Yan Jiu. 2004 Nov;33(6):710-2. [Study on the teratogenicity effects of genetically modified rice with Xa21 on rats] [Article in Chinese] Li Y, Piao J, Zhuo Q, Chen X, Mao D, Yang L, Yang X. National Institute for Nutrition and Food Safety, Chinese Center for Disease Control and Prevention, Beijing 100050, China. OBJECTIVE: To observe the effects of genetically modified rice with Xa21 on the development of rat embryos. METHODS: According to sex, weanling rats were divided into four groups: transgenic rice group, non-transgenic rice group, AIN93G negative control group and MATDA positive control group. The rats were fed with corresponding food for 90 days and mated. The development of maternal rats and embryos were observed. RESULTS: The body weight gain of pregnant rats and the body weight, body length and tail length of fetal rats in transgenic rice group were significant increased than those in positive control group. The number of death embryos and adsorption embryos, the malformation rate (appearance, viscera, skeleton) in transgenic rice group were lower than those in positive control group. There were no significant difference of all indicators among transgenic rice group, non-transgenic rice group and AIN93G negative control group. CONCLUSION: Compared with the non-transgenic rice, transgenic rice modified with Xa21 gene showed no significant differences in rat pregnant rate and embryo development. Publication Types: English Abstract Research Support, Non-U.S. Gov't PMID: 15727184 [PubMed - indexed for MEDLINE] 1072: Sci Eng Ethics. 2005 Jan;11(1):137-49. Property rights and genetic engineering: developing nations at risk. Shrader-Frechette K. Department of Philosophy and Department of Biological Sciences, 100 Malloy Hall, University of Notre Dame, Notre Dame, IN 46556, USA. Kristin.Shrader-Frechette.1@nd.edu Eighty percent of (commercial) genetically engineered seeds (GES) are designed only to resist herbicides. Letting farmers use more chemicals, they cut labor costs. But developing nations say GES cause food shortages, unemployment, resistant weeds, and extinction of native cultivars when "volunteers" drift nearby. While GES patents are reasonable, this paper argues many patent policies are not. The paper surveys GE technology, outlines John Locke's classic account of property rights, and argues that current patent policies must be revised to take account of Lockean ethical constraints. After answering a key objection, it provides concrete suggestions for implementing its ethical conclusions. PMID: 15727008 [PubMed - indexed for MEDLINE] 1073: Br J Cancer. 2005 Mar 14;92(5):873-5. Mutagenicity of comfrey (Symphytum Officinale) in rat liver. Mei N, Guo L, Fu PP, Heflich RH, Chen T. Division of Genetic and Reproductive Toxicology, National Center for Toxicological Research, US Food and Drug Administration, HFT-130, 3900 NCTR Road, Jefferson, AR 72079, USA. Comfrey is a rat liver toxin and carcinogen that has been used as a vegetable and herbal remedy by humans. In order to evaluate the mechanisms underlying its carcinogenicity, we examined the mutagenicity of comfrey in the transgenic Big Blue rat model. Our results indicate that comfrey is mutagenic in rat liver and the types of mutations induced by comfrey suggest that its tumorigenicity results from the genotoxicity of pyrrolizidine alkaloids in the plant. Publication Types: In Vitro Research Support, Non-U.S. Gov't PMID: 15726100 [PubMed - indexed for MEDLINE] 1074: J Agric Environ Ethics. 1999;11(3):197-217. Ethical issues in livestock cloning. Thompson PB. Department of Philosophy, Purdue University, West Lafayette, IN 47907-1360, USA. pault@purdue.edu Although cloning may eventually become an important technology for livestock production, four ethical issues must be addressed before the practice becomes widespread. First, researchers must establish that the procedure is not detrimental to the health or well-being of affected animals. Second, animal research institutions should evaluate the net social benefits to livestock producers by weighing the benefits to producers against the opportunity cost of research capacity lost to biomedical projects. Third, scientists should consider the indirect effects of cloning research on the larger ethical issues surrounding human cloning. Finally, the market structure for products of cloned animals should protect individual choice, and should recognize that many individuals find the prospect of cloning (or consuming cloned animals) repugnant. Analysis of these four issues is complicated by spurious arguments alleging that cloning will have a negative impact on environment and genetic diversity. PMID: 15719505 [PubMed - indexed for MEDLINE] 1075: Eur J Histochem. 2004 Oct-Dec;48(4):448-54. Ultrastructural analysis of testes from mice fed on genetically modified soybean. Vecchio L, Cisterna B, Malatesta M, Martin TE, Biggiogera M. We have considered the possible effects of a diet containing genetically modified (GM) soybean on mouse testis. This organ, in fact, is a well known bioindicator and it has already been utilized, for instance, to monitor pollution by heavy metals. In this preliminary study, we have focussed our attention on Sertoli cells, spermatogonia and spermatocytes by means of immunoelectron microscopy. Our results point out that the immunolabelling for Sm antigen, hnRNPs, SC35 and RNA Polymerase II is decreased in 2 and 5 month-old GM-fed mice, and is restored to normal at 8 months. In GM-fed mice of all ages considered, the number of perichromatin granules is higher and the nuclear pore density lower. Moreover, we found enlargements in the smooth endoplasmic reticulum in GM-fed mice Sertoli cells. A possible role played by traces of the herbicide to which the soybean is resistant is discussed. Publication Types: Letter Research Support, Non-U.S. Gov't PMID: 15718213 [PubMed - indexed for MEDLINE] 1076: J Agric Food Chem. 2005 Feb 23;53(4):912-8. A microarray platform for parallel detection of five transgenic events in foods: a combined polymerase chain reaction-ligation detection reaction-universal array method. Bordoni R, Germini A, Mezzelani A, Marchelli R, De Bellis G. Istituto di Tecnologie Biomediche, Consiglio Nazionale delle Ricerche, Via Fratelli Cervi 93, I-20090 Segrate (MI), Italy. We recently developed a multiplex polymerase chain reaction (PCR) system for the simultaneous detection of four transgenic maize (MON810, Bt176, Bt11, and GA21), one transgenic soybean (Roundup Ready), and two control genes (lectin and zein). Because PCR can lead to ambiguous interpretations due to low specificity, we have developed the ligation detection reaction (LDR) combined with a universal array as a molecular tool to confirm results of PCR analysis. Here, we describe the PCR-LDR-universal array procedure and demonstrate its specificity in revealing the presence of transgenic DNA in experimental samples, raw materials, and commercial foodstuffs. Publication Types: Research Support, Non-U.S. Gov't PMID: 15712997 [PubMed - indexed for MEDLINE] 1077: McGeorge Law Rev. 2000 Fall;32(1):89-110. Biotechnology and the creation of ethics. Coletta RR. University of the Pacific, McGeorge School of Law, USA. PMID: 15709268 [PubMed - indexed for MEDLINE] 1078: Mutat Res. 2005 Mar 1;570(2):205-14. The effect of UDP-glucuronosyltransferase 1A1 expression on the mutagenicity and metabolism of the cooked-food carcinogen 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine in CHO cells. Malfatti MA, Wu RW, Felton JS. Biology and Biotechnology Research Program, Lawrence Livermore National Laboratory, P.O. Box 808, L-452, Livermore, CA 94551-9900, USA. UDP-glucuronosyltransferase proteins (UGT) catalyze the glucuronidation of both endogenous and xenobiotic compounds. In previous studies, UGT1A1 has been implicated in the detoxification of certain food-borne carcinogenic-heterocyclic amines. To determine the importance of UDP-glucuronosyltransferase 1A1 (UGT1A1) in the biotransformation of the cooked-food carcinogen 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP), genetically modified CHO cells that are nucleotide excision repair-deficient, and express cytochrome P4501A2 (UV5P3 cell line) were transfected with a cDNA plasmid of human UGT1A1 to establish the UDP-glucuronosyltransferase 1A1 expressing 5P3hUGT1A1 cell line. Expression of the UGT1A1 gene was verified by screening neo gene expressing clonal isolates (G-418 resistant) for their sensitivity to cell killing from PhIP exposure. Five of 11 clones were chosen for further analysis due to their resistance to cell killing. Western blot analysis was used to confirm the presence of the UGT1A1 and CYP1A2 proteins. All five clones displayed a 52-kDa protein band, which corresponded to a UGT1A1 control protein. Only four of the clones had a protein band that corresponded to the CYP1A2 control protein. Correct fragment size of the cDNAs in the remaining four clones was confirmed by RT-PCR and quantification of the mRNA product was accomplished by real-time RT-PCR. Expression of UGT1A1 in the transfected cells was 10(4)-10(5)-fold higher relative to the UV5P3 parental cells. One clone (#14) had a 10-fold higher increase in expression at 1.47 x 10(5) over the other three clones. This clone was also the most active in converting N-hydroxy-PhIP to N-hydroxy-PhIP glucuronide conjugates in microsomal metabolism assays. Based on the D50 values, the cytotoxic effect of PhIP was decreased approximately 350-fold in the 5P3hUGT1A1 cells compared to the UV5P3 control cells. In addition, no significant increase in mutation frequency was observed in the transfected cells. These results clearly indicate that UGT1A1 plays a critical role in PhIP biotransformation, providing protection against PhIP-mediated cytotoxicity and mutagenicity. Publication Types: Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S. PMID: 15708579 [PubMed - indexed for MEDLINE] 1079: Regul Toxicol Pharmacol. 2005 Mar;41(2):134-49. Epub 2005 Jan 18. Safety evaluation of the phosphinothricin acetyltransferase proteins encoded by the pat and bar sequences that confer tolerance to glufosinate-ammonium herbicide in transgenic plants. Hérouet C, Esdaile DJ, Mallyon BA, Debruyne E, Schulz A, Currier T, Hendrickx K, van der Klis RJ, Rouan D. Bayer CropScience SA, Sophia Antipolis, France. corinne.herouet@bayercropscience.com Transgenic plant varieties, which are tolerant to glufosinate-ammonium, were developed. The herbicide tolerance is based upon the presence of either the bar or the pat gene, which encode for two homologous phosphinothricin acetyltransferases (PAT), in the plant genome. Based on both a review of published literature and experimental studies, the safety assessment reviews the first step of a two-step-approach for the evaluation of the safety of the proteins expressed in plants. It can be used to support the safety of food or feed products derived from any crop that contains and expresses these PAT proteins. The safety evaluation supports the conclusion that the genes and the donor microorganisms (Streptomyces) are innocuous. The PAT enzymes are highly specific and do not possess the characteristics associated with food toxins or allergens, i.e., they have no sequence homology with any known allergens or toxins, they have no N-glycosylation sites, they are rapidly degraded in gastric and intestinal fluids, and they are devoid of adverse effects in mice after intravenous administration at a high dose level. In conclusion, there is a reasonable certainty of no harm resulting from the inclusion of the PAT proteins in human food or in animal feed. PMID: 15698537 [PubMed - indexed for MEDLINE] 1080: Nat Biotechnol. 2005 Feb;23(2):170. Comment on: Nat Biotechnol. 2004 Dec;22(12):1503-5. Pharmacrops and bioterror. Wuerthele S. Publication Types: Comment Letter PMID: 15696140 [PubMed - indexed for MEDLINE] 1081: Proc Biol Sci. 2005 Jan 22;272(1559):111-9. Management of genetically modified herbicide-tolerant sugar beet for spring and autumn environmental benefit. May MJ, Champion GT, Dewar AM, Qi A, Pidgeon JD. Broom's Barn Research Station, Higham, Bury St Edmunds, Suffolk IP28 6NP, UK. mike.may@bbsrc.ac.uk When used in genetically modified herbicide-tolerant (GMHT) crops, glyphosate provides great flexibility to manipulate weed populations with consequences for invertebrates and higher trophic levels, for example birds. A range of timings of band and overall spray treatments of glyphosate to GMHT sugar beet were compared with a conventional weed control programme in four field trials over 2 years. Single overall sprays applied between 200 and 250 accumulated day degrees (above a base air temperature of 3 degrees C; degrees Cd) and band applied treatments applied at 10% or 20% ground cover within the crop rows generally gave significantly greater weed biomass and seed rain than conventional treatments, while later band sprays (more than 650 degrees Cd) reduced seed return. Two overall sprays of glyphosate produced low weed biomass and generally lowest seed return of all treatments but tended to give some of the highest yields. However, the early overall sprays (200-250 degrees Cd) and band sprays gave as good or better yields than the conventional and were generally equivalent to the two overall-spray programme. Viable seeds in the soil after the experiment were generally higher following the early overall (200-250 degrees Cd) and the band spray treatments than following the conventional. The results show that altered management of GMHT sugar beet can provide alternative scenarios to those of the recent Farm Scale Evaluation trials. Without yield loss they can enhance weed seed banks and autumn bird food availability compared with conventional management, or provide early season benefits to invertebrates and nesting birds, depending on the system chosen. Conventional weed control does not have the flexibility to enable these scenarios that benefit both agriculture and environment, although there may be some options for increasing weed seed return in autumn. Publication Types: Research Support, Non-U.S. Gov't PMID: 15695200 [PubMed - indexed for MEDLINE] 1082: Public Health. 2005 Feb;119(2):75-6. Future imperfect. Mackie P, Sim F. Publication Types: Editorial PMID: 15694953 [PubMed - indexed for MEDLINE] 1083: Biotechnol Adv. 2005 Mar;23(2):93-6. Is the battle over genetically modified foods finally over? Saleh-Lakha S, Glick BR. Publication Types: Editorial PMID: 15694121 [PubMed - indexed for MEDLINE] 1084: Przegl Lek. 2004;61 Suppl 3:22-4. [Genetically modified food and allergy] [Article in Polish] Wiackowski SK. Katedra Ekologii i Ochrony Srodowiska, Akademia Swietokrzyska, Kielce. skwiack@wp.pl Author discusses both successes and threats related with introduction of new organisms to the natural environment. Attention was sacrificed not only profits but also different threat influencing environment and human health. Publication Types: English Abstract Review PMID: 15682936 [PubMed - indexed for MEDLINE] 1085: J AOAC Int. 2004 Nov-Dec;87(6):1466-74. Effects of chemical, physical, and technological processes on the nature of food allergens. Poms RE, Anklam E. European Commission, Joint Research Centre, Institute for Reference Materials and Measurements, Retieseweg, 2440 Geel, Belgium. A review is presented of studies of different processing techniques and their effect on the allergenicity and antigenicity of certain allergenic foods. An overview of investigated technologies is given with regard to their impact on the protein structure and their potential application in the production of hypoallergenic foods. The use of physical processes (such as heating, high pressure, microparticulation, ultrafiltration, and irradiation), chemical processes (such as proteolysis, fermentation, and refining by extraction), and biotechnological approaches, as well as the effects of these processes on individual allergenic foods, are included. Additionally, the implications of food processing for food allergen analysis with respect to food safety assessment and industrial quality control are briefly discussed. Publication Types: Review PMID: 15675460 [PubMed - indexed for MEDLINE] 1086: J AOAC Int. 2004 Nov-Dec;87(6):1423-32. Assessing the allergenicity of proteins introduced into genetically modified crops using specific human IgE assays. Goodman RE, Leach JN. Monsanto Co, St Louis, MO 63167, USA. rgoodman2@unlnotes.unl.edu Global commercial production of genetically modified (GM) crops has grown to over 67 million hectares annually, primarily of herbicide-tolerant and insect protection crop varieties. GM crops are produced by the insertion of specific genes that either encode a protein, or a regulatory RNA sequence. A comprehensive safety evaluation is conducted for each new commercial GM crop, including an assessment of the potential allergenicity of any newly introduced protein. If the gene was derived from an allergenic organism, or the protein sequence is highly similar to a known allergen, immunoassays, e.g., Western blot assays and enzyme-linked immunosorbent assay tests, are performed to identify protein-specific IgE binding by sera of individuals allergic to the gene source, or the source of the sequence-matched allergen. Although such assays are commonly used to identify previously unknown allergens, criteria have not been established to demonstrate that a protein is unlikely to cause allergic reactions. This review discusses factors that affect the predictive value of these tests, including clinical selection criteria for serum donors, selection of blocking reagents to reduce nonspecific antibody binding, inhibition assays to verify specificity of binding, and scientifically justified limits of detection (sensitivity) in the absence of information regarding biological thresholds. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 15675455 [PubMed - indexed for MEDLINE] 1087: J AOAC Int. 2004 Nov-Dec;87(6):1342-55. Real-time polymerase chain reaction-based approach for quantification of the pat gene in the T25 Zea mays event. Weighardt F, Barbati C, Paoletti C, Querci M, Kay S, De Beuckeleer M, Van den Eede G. European Commission, Joint Research Centre, Institute for Health and Consumer Protection (JRC-IHCP), Via Enrico Fermi, 1, I-21020 Ispra (VA), Italy. In Europe, a growing interest for reliable techniques for the quantification of genetically modified component(s) of food matrixes is arising from the need to comply with the European legislative framework on novel food products. Real-time polymerase chain reaction (PCR) is currently the most powerful technique for the quantification of specific nucleic acid sequences. Several real-time PCR methodologies based on different molecular principles have been developed for this purpose. The most frequently used approach in the field of genetically modified organism (GMO) quantification in food or feed samples is based on the 5'-3'-exonuclease activity of Taq DNA polymerase on specific degradation probes (TaqMan principle). A novel approach was developed for the establishment of a TaqMan quantification system assessing GMO contents around the 1% threshold stipulated under European Union (EU) legislation for the labeling of food products. The Zea mays T25 elite event was chosen as a model for the development of the novel GMO quantification approach. The most innovative aspect of the system is represented by the use of sequences cloned in plasmids as reference standards. In the field of GMO quantification, plasmids are an easy to use, cheap, and reliable alternative to Certified Reference Materials (CRMs), which are only available for a few of the GMOs authorized in Europe, have a relatively high production cost, and require further processing to be suitable for analysis. Strengths and weaknesses of the use of novel plasmid-based standards are addressed in detail. In addition, the quantification system was designed to avoid the use of a reference gene (e.g., a single copy, species-specific gene) as normalizer, i.e., to perform a GMO quantification based on an absolute instead of a relative measurement. In fact, experimental evidences show that the use of reference genes adds variability to the measurement system because a second independent real-time PCR-based measurement must be performed. Moreover, for some reference genes no sufficient information on copy number in and among genomes of different lines is available, making adequate quantification difficult. Once developed, the method was subsequently validated according to IUPAC and ISO 5725 guidelines. Thirteen laboratories from 8 EU countries participated in the trial. Eleven laboratories provided results complying with the predefined study requirements. Repeatability (RSDr) values ranged from 8.7 to 15.9%, with a mean value of 12%. Reproducibility (RSDR) values ranged from 16.3 to 25.5%, with a mean value of 21%. Following Codex Alimentarius Committee guidelines, both the limits of detection and quantitation were determined to be <0.1%. Publication Types: Multicenter Study PMID: 15675446 [PubMed - indexed for MEDLINE] 1088: Plant Foods Hum Nutr. 2004 Winter;59(1):35-44. Nixtamalized flour from quality protein maize (Zea mays L). optimization of alkaline processing. Milán-Carrillo J, Gutiérrez-Dorado R, Cuevas-Rodríguez EO, Garzón-Tiznado JA, Reyes-Moreno C. Maestría en Ciencia y Tecnología de Alimentos, Facultad de Ciencias Químico Biológicas, Universidad Autónoma de Sinaloa, Sinaloa, México. Quality of maize proteins is poor, they are deficient in the essential amino acids lysine and tryptophan. Recently, in Mexico were successfully developed nutritionally improved 26 new hybrids and cultivars called quality protein maize (QPM) which contain greater amounts of lysine and tryptophan. Alkaline cooking of maize with lime (nixtamalization) is the first step for producing several maize products (masa, tortillas, flours, snacks). Processors adjust nixtamalization variables based on experience. The objective of this work was to determine the best combination of nixtamalization process variables for producing nixtamalized maize flour (NMF) from QPM V-537 variety. Nixtamalization conditions were selected from factorial combinations of process variables: nixtamalization time (NT, 20-85 min), lime concentration (LC, 3.3-6.7 g Ca(OH)2/l, in distilled water), and steep time (ST, 8-16 hours). Nixtamalization temperature and ratio of grain to cooking medium were 85 degrees C and 1:3 (w/v), respectively. At the end of each cooking treatment the steeping started for the required time. Steeping was finished by draining the cooking liquor (nejayote). Nixtamal (alkaline-cooked maize kernels) was washed with running tap water. Wet nixtamal was dried (24 hours, 55 degrees C) and milled to pass through 80-US mesh screen to obtain NMF. Response surface methodology (RSM) was applied as optimization technique, over four response variables: In vitro protein digestibility (PD), total color difference (deltaE), water absorption index (WAI), and pH. Predictive models for response variables were developed as a function of process variables. Conventional graphical method was applied to obtain maximum PD, WAI and minimum deltaE, pH. Contour plots of each of the response variables were utilized applying superposition surface methodology, to obtain three contour plots for observation and selection of best combination of NT (31 min), LC (5.4 g Ca(OH)2/l), and ST (8.1 hours) for producing optimized NMF from QPM. Publication Types: Research Support, Non-U.S. Gov't PMID: 15675150 [PubMed - indexed for MEDLINE] 1089: Transgenic Res. 2004 Dec;13(6):583-91. Re-interpreting some common objections to three transgenic applications: GM foods, xenotransplantation and germ line gene modification (GLGM). Carter L. Office of Public Policy and Ethics, Institute for Molecular Bioscience, The University of Queensland, Australia. l.carter@imb.uq.edu.au Concerns about safety to the individual, the wider community and the potential impact on the environment are typical consequentialist objections to transgenesis that feature prominently in public debates about its ethical acceptability. I consider some of these claims with respect to their motivation, validity and their overall influence on public policy using three well-discussed applications of transgenesis: GM foods, xenotransplantation and germ line gene modification (GLGM). Publication Types: Review PMID: 15672839 [PubMed - indexed for MEDLINE] 1090: Risk Anal. 2004 Dec;24(6):1515-27. Elicitation of expert judgments of uncertainty in the risk assessment of herbicide-tolerant oilseed crops. Krayer von Krauss MP, Casman EA, Small MJ. Department of Environment and Resources, Technical University of Denmark, Lyngby, Denmark. mkk@er.dtu.dk One of the lay public's concerns about genetically modified (GM) organisms (GMO) and related emerging technologies is that not all the important risks are evaluated or even identified yet--and that ignorance of the unanticipated risks could lead to severe environmental or public health consequences. To some degree, even the scientists who participated in the analysis of the risks from GMOs (arguably the people most qualified to critique these analyses) share some of this concern. To formally explore the uncertainty in the risk assessment of a GM crop, we conducted detailed interviews of seven leading experts on GM oilseed crops to obtain qualitative and quantitative information on their understanding of the uncertainties associated with the risks to agriculture from GM oilseed crops (canola or rapeseed). The results of these elicitations revealed three issues of potential concern that are currently left outside the scope of risk assessments. These are (1) the potential loss of the agronomic and environmental benefits of glyphosate (a herbicide widely used in no-till agriculture) due to the combined problems of glyphosate-tolerant canola and wheat volunteer plants, (2) the growing problem of seed lot contamination, and (3) the potential market impacts. The elicitations also identified two areas where knowledge is insufficient. These are: the occurrence of hybridization between canola and wild relatives and the ability of the hybrids to perpetuate themselves in nature, and the fate of the herbicide-tolerance genes in soil and their interaction with soil microfauna and -flora. The methodological contribution of this work is a formal approach to analyzing the uncertainty surrounding complex problems. Publication Types: Research Support, Non-U.S. Gov't PMID: 15660608 [PubMed - indexed for MEDLINE] 1091: Risk Anal. 2004 Dec;24(6):1475-86. Trust, the asymmetry principle, and the role of prior beliefs. Poortinga W, Pidgeon NF. Centre for Environmental Risk, University of East Anglia, Norwich NR4 7TJ, UK. w.poortinga@uea.ac.uk Within the risk literature there is an ongoing debate on whether trust is vulnerable or enduring. Previous research on nuclear energy by Slovic in 1993 has shown that negative events have much greater impact on self-reported trust than do positive events. Slovic attributes this to the asymmetry principle: specifically, that trust is much easier to destroy than to create. In a questionnaire survey concerning genetically modified (GM) food in Britain (n= 396) we similarly find that negative events have a greater impact on trust than positive events. Because public opinion in Britain is skewed in the direction of opposition toward GM food, the pattern of results could either be caused by the fact that negative information is more informative than positive information (a negativity bias) or reflect the influence of people's prior attitudes toward the issue (a confirmatory bias). The results were largely in line with the confirmatory bias hypothesis: participants with clear positive or negative beliefs interpreted events in line with their existing attitude position. However, for participants with intermediate attitudes, negative items still had greater impact than the positive. This latter finding suggests that, congruent with the negativity bias hypothesis, negative information may still be more informative than positive information for undecided people. The study also identified the labeling of GM products, consulting the public, making biotechnology companies liable for any damage, and making a test available to detect GM produce as being particularly important preconditions for maintaining trust in the regulation of agricultural biotechnology. Publication Types: Research Support, Non-U.S. Gov't PMID: 15660605 [PubMed - indexed for MEDLINE] 1092: J Appl Microbiol. 2005;98(2):418-28. High level expression of a recombinant acid phytase gene in Pichia pastoris. Xiong AS, Yao QH, Peng RH, Han PL, Cheng ZM, Li Y. Agro-Biotechnology Research Center of Shanghai Academy of Agricultural Sciences, Shanghai, China. AIMS: To achieve high phytase yield with improved enzymatic activity in Pichia pastoris. METHODS AND RESULTS: The 1347-bp phytase gene of Aspergillus niger SK-57 was synthesized using a successive polymerase chain reaction and was altered by deleting intronic sequences, optimizing codon usage and replacing its original signal sequence with a synthetic signal peptide (designated MF4I) that is a codon-modified Saccharomyces cerevisiae mating factor alpha-prepro-leader sequence. The gene constructs containing wild type or modified phytase gene coding sequences under the control of the highly-inducible alcohol oxidase gene promoter with the MF4I- or wild type alpha-signal sequence were used to transform Pichia pastoris. The P. pastoris strain that expressed the modified phytase gene (phyA-sh) with MF4I sequence produced 6.1 g purified phytase per litre of culture fluid, with the phytase activity of 865 U ml(-1). The expressed phytase varied in size (64, 67, 87, 110 and 120 kDa), but could be deglycosylated to produce a homogeneous 64 kDa protein. The recombinant phytase had two pH optima (pH 2.5 and pH 5.5) and an optimum temperature of 60 degrees C. CONCLUSIONS: The P. pastoris strain with the genetically engineered phytase gene produced 6.1 g l(-1) of phytase or 865 U ml(-1) phytase activity, a 14.5-fold increase compared with the P. pastoris strain with the wild type phytase gene. SIGNIFICANCE AND IMPACT OF THE STUDY: The P. pastoris strain expressing the modified phytase gene with the MF4I signal peptide showed great potential as a commercial phytase production system. Publication Types: Research Support, Non-U.S. Gov't PMID: 15659196 [PubMed - indexed for MEDLINE] 1093: J Agric Food Chem. 2005 Jan 26;53(2):183-90. Validation of a tomato-specific gene, LAT52, used as an endogenous reference gene in qualitative and real-time quantitative PCR detection of transgenic tomatoes. Yang L, Pan A, Jia J, Ding J, Chen J, Cheng H, Zhang C, Zhang D. School of Life Science and Biotechnology, Shanghai Jiao Tong University, 800 Dongchuan Road, Shanghai 200240, People's Republic of China. Toward the development of reliable qualitative and quantitative Polymerase Chain Reaction (PCR) detection methods of transgenic tomatoes, one tomato (Lycopersicon esculentum) species specific gene, LAT52, was selected and validated as suitable for using as an endogenous reference gene in transgenic tomato PCR detection. Both qualitative and quantitative PCR methods were assayed with 16 different tomato varieties, and identical amplified products or fluorescent signals were obtained with all of them. No amplified products and fluorescent signals were observed when DNA samples from 20 different plants such as soybean, maize, rapeseed, rice, and Arabidopsis thaliana were used as templates. These results demonstrated that the amplified LAT52 DNA sequence was specific for tomato. Furthermore, results of Southern blot showed that the LAT52 gene was a single-copy gene in the different tested tomato cultivars. In qualitative and quantitative PCR analysis, the detection sensitivities were 0.05 and 0.005 ng of tomato genomic DNA, respectively. In addition, two real-time assays employing this gene as an endogenous reference gene were established, one for the quantification of processed food samples derived from nontransgenic tomatoes that contained degraded target DNA and the other for the quantification of the junction region of CaMV35s promoter and the anti-sense ethylene-forming enzyme (EFE) gene in transgenic tomato Huafan No. 1 samples. All of these results indicated that the LAT52 gene could be successfully used as a tomato endogenous reference gene in practical qualitative and quantitative detection of transgenic tomatoes, even for some processed foods derived from transgenic and nontransgenic tomatoes. Publication Types: Research Support, Non-U.S. Gov't PMID: 15656646 [PubMed - indexed for MEDLINE] 1094: Soc Sci Med. 2005 Apr;60(7):1603-12. Psychosocial and cultural factors affecting the perceived risk of genetically modified food: an overview of the literature. Finucane ML, Holup JL. Center for Health Research, Hawai'i Kaiser Permanente Hawai'i, 501 Alakawa Street, Suite 201, Honolulu, Hawai'i 96817, USA. melissa.l.finucane@kp.org The rapid globalization of the world economy has increased the need for an astute understanding of cultural differences in perceptions, values, and ways of thinking about new food technologies. In this paper, we describe how socio-psychological and cultural factors may affect public perceptions of the risk of genetically modified (GM) food. We present psychological, sociological, and anthropological research on risk perception as a framework for understanding cross-national differences in reactions to GM food. Differences in the cultural values and circumstances of people in the US, European countries, and the developing world are examined. The implications of cultural theory for risk communication and decision making about GM food are discussed and directions for future research highlighted. Publication Types: Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Review PMID: 15652691 [PubMed - indexed for MEDLINE] 1095: Psychopharmacology (Berl). 2005 Jun;179(4):854-62. Epub 2005 Jan 12. Complex discriminative stimulus properties of (+)lysergic acid diethylamide (LSD) in C57Bl/6J mice. Benneyworth MA, Smith RL, Barrett RJ, Sanders-Bush E. Department of Pharmacology, Vanderbilt University School of Medicine, 8148 MRB III, Nashville, TN 37232, USA. RATIONALE: The drug discrimination procedure is the most frequently used in vivo model of hallucinogen activity. Historically, most drug discrimination studies have been conducted in the rat. With the development of genetically modified mice, a powerful new tool has become available for investigating the mechanisms of drug-induced behavior. The current paper is part of an ongoing effort to determine the utility of the drug discrimination technique for evaluating hallucinogenic drugs in mice. OBJECTIVE: To establish the training procedures and characterize the stimulus properties of (+)lysergic acid diethylamide (LSD) in mice. METHODS: Using a two-lever drug discrimination procedure, C57Bl/6J mice were trained to discriminate 0.45 mg/kg LSD vs saline on a VI30 sec schedule of reinforcement, with vanilla-flavored Ensure serving as the reinforcer. RESULTS: As in rats, acquisition was orderly, but the training dose was nearly five-fold higher for mice than rats. LSD lever selection was dose-dependent. Time-course studies revealed a rapid loss of the LSD stimulus effects. The 5-HT(2A/2C) receptor agonist, 2,5-dimethoxy-4-bromoamphetamine [(-)DOB] (1.0 mg/kg), substituted fully for LSD and the 5-HT(1A) receptor agonist, 8-hydroxy-2-(di-n-propylamino)-tetralin (8-OH-DPAT) (1.6 mg/kg), substituted partially for LSD. Pretreatment with the 5-HT(2A) receptor-selective antagonist, MDL 100907, or the 5-HT(1A)-selective antagonist WAY 100635, showed that each antagonist only partially blocked LSD discrimination. Substitution of 1.0 mg/kg (-)DOB for LSD was fully blocked by pretreatment with MDL 100907 but unaltered by WAY 100635 pretreatment. CONCLUSIONS: These data suggest that in mice the stimulus effects of LSD have both a 5-HT(2A) receptor and a 5-HT(1A) receptor component. Publication Types: Research Support, N.I.H., Extramural Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S. PMID: 15645221 [PubMed - indexed for MEDLINE] 1096: J Anim Sci. 2005 Feb;83(2):400-7. Nutrient digestibility in sheep fed diets containing Roundup Ready or conventional fodder beet, sugar beet, and beet pulp. Hartnell GF, Hvelplund T, Weisbjerg MR. Monsanto Co., St. Louis, MO 63167, USA. gary.f.hartnell@monsanto.com The objective of this digestibility assessment was to determine whether there are significant differences in the digestibility of Roundup Ready (glyphosate-tolerant) and conventional sugar beet, fodder beet, and beet pulp produced from sugar beet varieties when fed to sheep (seven wethers per treatment group). Three experiments were conducted in this assessment. Experiment 1 (35 wethers) compared one glyphosate-tolerant fodder beet variety with four conventional varieties, Exp. 2 (42 wethers) compared one glyphosate-tolerant sugar beet variety with five conventional varieties, and Exp. 3 (42 wethers) compared beet pulp derived from glyphosate-tolerant sugar beet with beet pulp from five European locations. The experimental phase consisted of a 2-wk preliminary period followed by a 1-wk collection period for Exp. 1 and 2, and a 1-wk preliminary period followed by a 1-wk digestibility collection period for Exp. 3. Diets were comprised of grass hay at 30, 30, and 20% of DM for Exp. 1, 2, and 3, respectively, with the balance being beet components. Urea and sodium sulfate were supplemented (8 and 2.9 g, respectively, for Exp. 1 and 2; and 6 g and 2.16 g, respectively, for Exp. 3) to supply sufficient dietary N and S. Each diet was fed to sheep (96 +/- 0.9 kg) in the three experiments to at or near maintenance energy levels. Treatment differences were considered significant at P < 0.05. Apparent digestibilities of DM, OM, CP, NDF, ADF, and DE for glyphosate-tolerant fodder and sugar beets did not differ from those for commercial fodder and sugar beets in Exp. 1 and 2. There were differences (P < 0.05) in DM, OM, CP, NDF, ADF, and DE digestibilities influenced by the different varieties of beet pulp in Exp. 3, but these were not unique to just the Roundup Ready sugar beet variety. Digestibilities and feeding values of Roundup Ready fodder beet, sugar beet, and beet pulp produced from Roundup Ready sugar beet varieties were not influenced by the introduction of the Roundup Ready trait compared with conventional varieties. Publication Types: Comparative Study Research Support, Non-U.S. Gov't PMID: 15644513 [PubMed - indexed for MEDLINE] 1097: Protein Expr Purif. 2005 Feb;39(2):189-98. Optimization of expression and purification of two biologically active chimeric fusion proteins that consist of human interleukin-13 and Pseudomonas exotoxin in Escherichia coli. Joshi BH, Puri RK. Laboratory of Molecular Tumor Biology, Division of Cellular and Gene Therapies, Office of Cellular, Tissue and Gene Therapies, Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, MD, USA. We have previously reported that a variety of solid human tumor cell lines express a large number of receptors for interleukin-13 (IL-13). These receptors could be targeted with a chimeric fusion protein consisting of human IL-13 and a truncated form of Pseudomonas exotoxin (PE). We describe here optimization of critical steps involved in high yield expression of two recombinant chimeric fusion proteins for obtaining highly purified and biologically active cytotoxins in Escherichia coli. The chimeric constructs of human IL-13 and two 38 kDa truncated PEs: (i) PE38 and (ii) PE38QQR, (three lysine residues in PE38 at 590, 606, and 613 substituted with two glutamine and one arginine) were used for protein expression in pET prokaryotic expression vector system with kanamycin as a selection antibiotic. Our results suggest that fresh transformation of E. coli and induction by isopropyl-beta-D-thiogalactopyranoside (IPTG) for 6 h resulted in maximum protein expression. To further improve the yield, we used a genetically modified E. coli strain, BL21(DE3)pLysS, which carries a plasmid for lysozyme with a weak promoter that inhibits T7 RNA polymerase and minimizes protein production in the absence of IPTG. Use of this strain eliminated the need for lysozyme digestion of the induced bacteria to release inclusion bodies, which resulted in expression of purer protein as compared to the conventional BL21(DE3) strain. Additional protocol optimizations included 16 h solubilization of inclusion bodies, constitution of refolding buffer, and timing of dialysis. These proteins were finally purified by Q-Sepharose, mono-Q, and gel filtration chromatography. Between 14-22 and 21-28 mg highly purified and biologically active protein was obtained from 1L of BL21 (DE3) and BL21 (DE3) pLysS bacteria culture, respectively. As IL-13R targeting for brain tumor therapy offers an exciting treatment option, optimization of production of IL-13PE will enhance production of clinical grade material for Phase III clinical trials. PMID: 15642470 [PubMed - indexed for MEDLINE] 1098: J Biol Chem. 2005 Mar 18;280(11):10435-43. Epub 2005 Jan 7. A Dictyostelium mutant with reduced lysozyme levels compensates by increased phagocytic activity. Müller I, Subert N, Otto H, Herbst R, Rühling H, Maniak M, Leippe M. Department of Cell Biology, Kassel University, Heinrich-Plett-Strasse 40, 34132 Kassel, Germany. Lysozymes are bacteria-degrading enzymes and play a major role in the immune defense of animals. In free-living protozoa, lysozyme-like proteins are involved in the digestion of phagocytosed bacteria. Here, we purified a protein with lysozyme activity from Dictyostelium amoebae, which constitutes the founding member, a novel class of lysozymes. By tagging the protein with green fluorescent protein or the Myc epitope, a new type of lysozyme-containing vesicle was identified that was devoid of other known lysosomal enzymes. The most highly expressed isoform, encoded by the alyA gene, was knocked out by homologous recombination. The mutant cells had greatly reduced enzymatic activity and grew inefficiently when bacteria were the sole food source. Over time the mutant gained the ability to internalize bacteria more efficiently, so that the defect in digestion was compensated by increased uptake of food particles. Publication Types: Research Support, Non-U.S. Gov't PMID: 15640146 [PubMed - indexed for MEDLINE] 1099: J Sep Sci. 2004 Dec;27(17-18):1551-6. DNA separation by capillary electrophoresis with hydrophilic substituted celluloses as coating and sieving polymers. Application to the analysis of genetically modified meals. Giovannoli C, Anfossi L, Tozzi C, Giraudi G, Vanni A. Department of Analytical Chemistry, Via P Giuria 5, University of Turin, 10125 Turin, Italy. cristina.giovannoli@unito.it A coating procedure based on the physical adsorption of hydroxypropyl cellulose onto the wall of a capillary column has been successfully used for the separation of DNA fragments up to 500 bp. The method uses a running Tris-phosphate-EDTA buffer containing 2-hydroxyethyl cellulose as sieving polymer. The separation procedure shows good reproducibility (measured as RSD%) for consecutive runs (<0.64), for different days (< 1.15) and capillaries (<2.15), short analysis times, and a long coating lifetime. Good reproducibility and efficiency are even achieved by performing the separation in the presence of additives such as ethidium bromide and mannitol. The method is applied to the detection of GMOs in soybean and maize meals with an accurate evaluation of the length of DNA sequences, previously amplified by polymerase chain reaction. Publication Types: Research Support, Non-U.S. Gov't PMID: 15638166 [PubMed - indexed for MEDLINE] 1100: Nat Biotechnol. 2005 Jan;23(1):27-33. Erratum in: Nat Biotechnol. 2005 Mar;23(3):366. Poorer nations turn to publicly developed GM crops. Cohen JI. International Food Policy Research Institute (IFPRI), Environment and Production Technology Division, 2033 K Street, NW, Washington, DC, USA. j.cohen@cgiar.org Publication Types: Research Support, Non-U.S. Gov't PMID: 15637614 [PubMed - indexed for MEDLINE] 1101: Appl Microbiol Biotechnol. 2005 Jul;68(1):75-81. Epub 2005 Jan 6. Cell surface display system for Lactococcus lactis: a novel development for oral vaccine. Raha AR, Varma NR, Yusoff K, Ross E, Foo HL. Department of Bioprocess Technology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, Serdang, 43400, Selangor, Malaysia. raha@fsb.upm.edu.my The food-grade Lactococcus lactis is a potential vector to be used as a live vehicle for the delivery of heterologous proteins for vaccine and pharmaceutical purposes. We constructed a plasmid vector pSVac that harbors a 255-bp single-repeat sequence of the cell wall-binding protein region of the AcmA protein. The recombinant plasmid was transformed into Escherichia coli and expression of the gene fragment was driven by the T7 promoter of the plasmid. SDS-PAGE showed the presence of the putative AcmA' fragment and this was confirmed by Western blot analysis. The protein was isolated and purified using a His-tag affinity column. When mixed with a culture of L. lactis MG1363, ELISA and immunofluorescence assays showed that the cell wall-binding fragment was anchored onto the outer surface of the bacteria. This indicated that the AcmA' repeat unit retained the active site for binding onto the cell wall surface of the L. lactis cells. Stability assays showed that the fusion proteins (AcmA/A1, AcmA/A3) were stably docked onto the surface for at least 5 days. The AcmA' fragment was also shown to be able to strongly bind onto the cell surface of naturally occurring lactococcal strains and Lactobacillus and, with less strength, the cell surface of Bacillus sphericus. The new system designed for cell surface display of recombinant proteins on L. lactis was evaluated for the expression and display of A1 and A3 regions of the VP1 protein of enterovirus 71 (EV71). The A1 and A3 regions of the VP1 protein of EV71 were cloned upstream to the cell wall-binding domains of AcmA protein and successfully expressed as AcmA/A1 and AcmA/A3. Whole-cell ELISA showed the successful display of VP1 protein epitopes of EV71 on the surface of L. lactis. The success of the anchoring system developed in this study for docking the A1 and A3 epitopes of VP1 onto the surface of L. lactis cells opens up the possibilities of peptide and protein display for not only Lactococcus but also for other gram-positive bacteria. This novel way of displaying epitopes on the cell surface of L. lactis and other related organisms should be very useful in the delivery of vaccines and other useful proteins. Publication Types: Research Support, Non-U.S. Gov't PMID: 15635459 [PubMed - indexed for MEDLINE] 1102: Plant Mol Biol. 2004 Nov;56(4):661-9. Engineering cyanogen synthesis and turnover in cassava (Manihot esculenta). Siritunga D, Sayre R. The Ohio state university, 43210, Columbus, OH 43210, USA,. Cassava is the major root crop for a quarter billion subsistence farmers in sub-Saharan Africa. It is valued for its ability to grow in adverse environments and the food security it provides. Cassava contains potentially toxic levels of cyanogenic glycosides (linamarin) which protect the plant from herbivory and theft. The cyanogens, including linamarin and its deglycosylated product, acetone cyanohydrin, can be efficiently removed from the root by various processing procedures. Short-cuts in processing, which may occur during famines, can result in only partial removal of cyanogens. Residual cyanogens in cassava foods may cause neurological disorders or paralysis, particularly in nutritionally compromised individuals. To address this problem and to further understand the function of cyanogenic glycosides in cassava, we have generated transgenic cassava in which cyanogenic glycoside synthesis has been selectively inhibited in leaves and roots by antisense expression of CYP79D1/D2 gene fragments. The CYP79D1/D2 genes encode two highly similar cytochrome P450s that catalyze the first-dedicated step in cyanogenic glycoside synthesis. Transgenic plants in which the expression of these genes was selectively inhibited in leaves had substantially reduced (60- 94% reduction) linamarin leaf levels. Surprisingly, these plants also had a greater than a 99% reduction in root linamarin content. In contrast, transgenic plants in which the CYP79D1/D2 transcripts were reduced to non-detectable levels in roots had normal root linamarin levels. These results demonstrate that linamarin synthesized in leaves is transported to the roots and accounts for nearly all of the root linamarin content. Importantly, transgenic plants having reduced leaf and root linamarin content were unable to grow in the absence of reduced nitrogen (NH3) . Cassava roots have previously been demonstrated to have an active cyanide assimilation pathway leading to the synthesis of amino acids. We propose that cyanide derived from linamarin is a major source of reduced nitrogen for cassava root protein synthesis. Disruption of linamarin transport from leaves in CYP79D1/D2 anti-sense plants prevents the growth of cassava roots in the absence of an alternate source of reduced nitrogen. An alternative strategy for reducing cyanogen toxicity in cassava foods is to accelerate cyanogenesis and cyanide volatilization during food processing. To achieve this objective, we have expressed the leaf-specific enzyme hydroxynitrile lyase (HNL) in roots. HNL catalyzes the breakdown of acetone cyanohydrin to cyanide. Expression of HNL in roots accelerated cyanogenesis by more than three-fold substantially reducing the accumulation of acetone cyanohydrin during processing relative to wild-type roots. Publication Types: Comparative Study PMID: 15630626 [PubMed - indexed for MEDLINE] 1103: Trends Biotechnol. 2005 Jan;23(1):17-21. Transgenes for tea? Heritage J. School of Biochemistry and Microbiology, University of Leeds, Leeds, LS2 9JT, UK. j.heritage@leeds.ac.uk So far, no compelling scientific evidence has been found to suggest that the consumption of transgenic or genetically modified (GM) plants by animals or humans is more likely to cause harm than is the consumption of their conventional counterparts. Despite this lack of scientific evidence, the economic prospects for GM plants are probably limited in the short term and there is public opposition to the technology. Now is a good time to address several issues concerning GM plants, including the potential for transgenes to migrate from GM plants to gut microbes or to animal or human tissues, the consequences of consuming GM crops, either as fresh plants or as silage, and the problems caused by current legislation on GM labelling and beyond. Publication Types: Review PMID: 15629853 [PubMed - indexed for MEDLINE] 1104: Zhi Wu Sheng Li Yu Fen Zi Sheng Wu Xue Xue Bao. 2004 Oct;30(5):517-22. [Overexpression of GST gene accelerates the growth of transgenic Arabidopsis under salt stress.] [Article in Chinese] Qi YC, Zhang SM, Wang LP, Wang MD, Zhang H. College of Biological Technique and Food Science, Henan Agricultural University, Zhengzhou 450002, China. yuanchengqi2003@163.com The Suaeda salsa glutathione s-transferase gene (GST) was inserted downstream of the 35S promoter in the plant expression vector pROK II and then was introduced into Arabidopsis thaliana by Agrobacterium tumefaciens through floral dip method. Transformants were selected for their ability to grow on medium containing kanamycin. The fact that the GST gene had been transferred into the Arabidopsis thaliana genome was confirmed by the PCR-Southern blotting analysis. After cultivation, independent homozygous transgenic lines were obtained after selection of T(3) progenies on MS medium containing kanamycin. The expression of the gene transferred into the Arabidopsis thaliana was confirmed by Northern blotting. During salt stress, analysis of total glutathione (both oxidized and reduced type) and biomass of transgenic and wild Arabidopsis. The biomass of transgenic lines (GT) was slightly but significantly greater than that of wild type line (WT), and levels of oxidized glutathione (GSSG) were significantly higher in transgenic lines than in wild type. Therefore, overexpression of GST can increase Arabidopsis growth under salt stress, and this effect can be caused by oxidation of the reduced glutathione (GSH ). Publication Types: English Abstract Research Support, Non-U.S. Gov't PMID: 15627705 [PubMed - indexed for MEDLINE] 1105: Theriogenology. 2005 Jan 15;63(2):283-99. Perspectives for artificial insemination and genomics to improve global swine populations. Gerrits RJ, Lunney JK, Johnson LA, Pursel VG, Kraeling RR, Rohrer GA, Dobrinsky JR. Biotechnology and Germplasm Laboratory, ANRI, BARC, ARS, USDA, Beltsville, MD, USA. Civilizations throughout the world continue to depend on pig meat as an important food source. Approximately 40% of the red meat consumed annually worldwide (94 million metric tons) is pig meat. Pig numbers (940 million) and consumption have increased consistent with the increasing world population (FAO 2002). In the past 50 years, research guided genetic selection and nutrition programs have had a major impact on improving carcass composition and efficiency of production in swine. The use of artificial insemination (AI) in Europe has also had a major impact on pig improvement in the past 35 years and more recently in the USA. Several scientific advances in gamete physiology and/or manipulation have been successfully utilized while others are just beginning to be applied at the production level. Semen extenders that permit the use of fresh semen for more than 5 days post-collection are largely responsible for the success of AI in pigs worldwide. Transfer of the best genetics has been enabled by use of AI with fresh semen, and to some extent, by use of AI with frozen semen over the past 25 years. Sexed semen, now a reality, has the potential for increasing the rate of genetic progress in AI programs when used in conjunction with newly developed low sperm number insemination technology. Embryo cryopreservation provides opportunities for international transport of maternal germplasm worldwide; non-surgical transfer of viable embryos in practice is nearing reality. While production of transgenic animals has been successful, the low level of efficiency in producing these animals and lack of information on multigene interactions limit the use of the technology in applied production systems. Technologies based on research in functional genomics, proteomics and cloning have significant potential, but considerable research effort will be required before they can be utilized for AI in pig production. In the past 15 years, there has been a coordinated worldwide scientific effort to develop the genetic linkage map of the pig with the goal of identifying pigs with genetic alleles that result in improved growth rate, carcass quality, and reproductive performance. Molecular genetic tests have been developed to select pigs with improved traits such as removal of the porcine stress (RYR1) syndrome, and selection for specific estrogen receptor (ESR) alleles. Less progress has been made in developing routine tests related to diseases. Major research in genomics is being pursued to improve the efficiency of selection for healthier pigs with disease resistance properties. The sequencing of the genome of the pig to identify new genes and unique regulatory elements holds great promise to provide new information that can be used in pig production. AI, in vitro embryo production and embryo transfer will be the preferred means of implementing these new technologies to enhance efficiency of pig production in the future. Publication Types: Lectures PMID: 15626400 [PubMed - indexed for MEDLINE] 1106: Med Mal Infect. 2004 Nov;34(11):522-9. [Impact of genetic modifications on infectious diseases] [Article in French] Houdebine LM. INRA, UMR Biologie du développement et de la reproduction, 78312 Jouy-en-Josas, France. houdebine@jouy.inra.fr Genetic engineering offers the theoretical possibility to transfer any natural or modified gene into any living organism. This generates new and diverse situations which may contribute to the spreading of infectious diseases or on the contrary to control them. Problems may theoretically come from uncontrolled genes providing resistance to antibiotics, from the activation of genomic retroviral sequences, from enhanced sensitivity of the organism to pathogens, as well as from the generation of mutated microorganisms with a higher pathogenecity. On the contrary, various genetic modifications may create organisms resistant to infectious diseases, generate safe and efficient recombinant vaccines, or provide patients with proteins which stimulate their defense mechanisms. The major impacts of genetic modifications in the development of infectious diseases or on the contrary in their eradication are analyzed in this article. Publication Types: English Abstract Review PMID: 15620056 [PubMed - indexed for MEDLINE] 1107: Anal Bioanal Chem. 2005 Jan;381(1):72-4. Reliable GMO analysis. Trapmann S, Emons H. European Commission, Joint Research Centre, Institute for Reference Materials and Measurements (IRMM), Retieseweg 111, Geel 2440, Belgium. stefanie.trapmann@cec.eu.int PMID: 15616785 [PubMed - indexed for MEDLINE] 1108: Crit Rev Food Sci Nutr. 2004;44(6):425-36. Detection of genetically modified organisms in foods by DNA amplification techniques. García-Cañas V, Cifuentes A, González R. Institute of Industrial Fermentations, CSIC, Madrid, Spain. In this article, the different DNA amplification techniques that are being used for detecting genetically modified organisms (GMOs) in foods are examined. This study intends to provide an updated overview (including works published till June 2002) on the principal applications of such techniques together with their main advantages and drawbacks in GMO detection in foods. Some relevant facts on sampling, DNA isolation, and DNA amplification methods are discussed. Moreover; these analytical protocols are discuissed from a quantitative point of view, including the newest investigations on multiplex detection of GMOs in foods and validation of methods. Publication Types: Research Support, Non-U.S. Gov't PMID: 15615426 [PubMed - indexed for MEDLINE] 1109: Poult Sci. 2004 Dec;83(12):2029-38. Evaluation of broiler performance when fed insect-protected, control, or commercial varieties of dehulled soybean meal. Kan CA, Hartnell GF. Animal Sciences Group of Wageningen UR, Nutrition and Food, 8200 AB Lelystad, The Netherlands. kees.kan@wur.nl We evaluated the nutritional value of broiler diets containing approximately 35% soybean meal from insect-protected soybean containing CrylAc protein, or from a similar nontransgenic control, or from 7 reference commercial soybean varieties. The feeding trial lasted 41 d, and each treatment consisted of 10 replicates of 1-d-old Ross 508 broilers (5 pens males and 5 pens females). Each pen contained 12 birds, and at d 13, birds were randomly removed until 9 birds remained. Body weight and feed intake were measured on pen basis at 41 d. At d 42, 4 broilers per pen were slaughtered. The carcasses were dissected, and cut-up yields were determined. Dry matter, protein, and fat contents of breast meat as well as shear force of breast meat were determined. The data were analyzed by ANOVA. The BW and feed conversion ratio at d 41 averaged 2,435 g and 1.52, respectively. There were no significant treatment x sex interactions. Data for final BW, feed conversion, carcass yield, and breast meat data were not different (P < 0.05) between broilers fed insect-protected and those fed commercial soybean meal varieties. Thus, insect-protected soybean meal was nutritionally equivalent to nongenetically modified soybean varieties when fed to broilers. Publication Types: Clinical Trial Randomized Controlled Trial PMID: 15615017 [PubMed - indexed for MEDLINE] 1110: Genet Mol Res. 2004 Sep 30;3(3):432-40. Reduction of non-digestible oligosaccharides in soymilk: application of engineered lactic acid bacteria that produce alpha-galactosidase. LeBlanc JG, Silvestroni A, Connes C, Juillard V, de Giori GS, Piard JC, Sesma F. Centro de Referencias para Lactobacilos (CERELA - CONICET), Chacabuco 145, (4000) Tucumán, Argentina. Human consumption of soy-derived products has been limited by the presence of non-digestible oligosaccharides (NDO), such as the alpha-galactooligosaccharides raffinose and stachyose. Most mammals, including man, lack pancreatic alpha-galactosidase (alpha-Gal), which is necessary for the hydrolysis of these sugars. However, such NDO can be fermented by gas-producing microorganisms present in the cecum and large intestine, which in turn can induce flatulence and other gastrointestinal disorders in sensitive individuals.The use of microorganisms expressing alpha-Gal is a promising solution to the elimination of NDO before they reach the large intestine. In the present study, lactic acid bacteria engineered to degrade NDO have been constructed and are being used as a tool to evaluate this solution. The alpha-Gal structural genes from Lactobacillus plantarum ATCC8014 (previously characterized in our laboratory) and from guar have been cloned and expressed in Lactococcus lactis. The gene products were directed to different bacterial compartments to optimize their possible applications. The alpha-Gal-producing strains are being evaluated for their efficiency in degrading raffinose and stachyose: i) in soymilk fermentation when used as starters and ii) in situ in the upper gastrointestinal tract when administered to animals orally, as probiotic preparations. The expected outcomes and possible complications of this project are discussed. Publication Types: Research Support, Non-U.S. Gov't PMID: 15614733 [PubMed - indexed for MEDLINE] 1111: Lett Appl Microbiol. 2005;40(1):37-43. Avoidance of oxidative-stress perturbation in yeast bioprocesses by proteomic and genomic biostrategies? Wiseman A. Molecular Toxicology Group, School of Biomedical & Life Sciences, University of Surrey, Guildford, Surrey, UK. Helen.Wiseman@kcl.ac.uk AIMS: Bioprocess oxidative stress caused by many reactive oxygen species (ROS) can lead to largely irreversible perturbation of yeast bioprocesses. These include the production of proteins derived from recombinant DNA yeast technology (aerobically grown Saccharomyces cerevisiae). These proteins include rennin, amyloglucosidases (glucamylases), interferons, interleukins, insulin, monoclonal antibodies, tissue plasminogen activators (t-PA), sexually transmitted disease antigens, and measles, mumps and rubella antigens, growth hormones, somatotropin, blood clotting factors VIII and XIII. In addition, there may be a demand for severe acute respiratory syndrome-coronavirus antigens, hepatitis A, B and C viral-selected antigens, HIV retroviral antigens, influenza antigens, trypanosomal antigens, and foot and mouth disease antigens. Prevention of oxidative stress has been achieved by application of antioxidant redox metalloenzymes such as superoxide dismutases (containing Cu/Zn cytosolic, Mn mitochondrial and Fe bacterial) glutathione peroxidases (and other Se-containing proteins and enzymes such as the thioredoxins), catalases (Fe-containing), cytochrome c peroxidases (Fe-containing), ceruloplasmins (Cu-containing), metallothionines (these cysteine thiol-rich proteins bind ions of cadmium and mercury) and tyrosinases(Cu-containing). METHODS AND RESULTS: ROS are generated inadvertently by single metal valency couples such as FeII/FeIII and by FeIII/FeV present in 2700 (including 57 human) isoforms in cytochromes P450 mixed-function oxidases (EC 1.14.14.1; O2 : mono-oxygenase NADPH/NADH requiring). In addition, mixed-metal couples such as valency unmatched forms in CuI/FeII and FeIII/MnIV can recycle electrons. Moreover, proteins/protein chaperone couples can recycle electrons, often where futile-recycling systems have been instigated. Furthermore, oxidized membrane phospholipids (R) can form ROOH (lipid hydroperoxides) and ROH (lipid alkoxides) that can generate ROS through Fenton chemistry (iron-catalysed) chain reactions. Utilization of chain-breaking antioxidants such as vitamin E (alpha-tocopherol) in the lipid phase and vitamin C (ascorbate) in the aqueous phase can terminate these ROS-producing reactions. CONCLUSIONS: The main significance of the study is that proteomic strategies of relief from bioprocess perturbation by ROS of yeast fermentations (used to manufacture proteins required in the food and therapeutic bioindustries) may become possible through addition of selected proteins (including metalloenzymes). The main impact of the study is that the utilization of genetically modified (GM) yeast produced by recombinant DNA technology genomic strategies could circumvent the bioprocessing problems that otherwise result from the bioprocess perturbations: this is as a result of oxidative stress caused by ROS, which is avoidable by deployment of appropriate antioxidants such as vitamins E, C and D (and antioxidant proteins and enzymes often of microbial origin via recombinant DNA technology). PMID: 15613000 [PubMed - indexed for MEDLINE] 1112: Environ Biosafety Res. 2004 Apr-Jun;3(2):99-107. A decade of European field trials with genetically modified plants. Lheureux K, Menrad K. European Commission, Directorate-General Joint Research Centre, Institute for Prospective Technological Studies, D-94315 Straubing, Germany. Karine.Lheureux@efsa.eu.int This article analyzes the development of notifications of genetically modified plants field trials in the European Union from 1991 to 2001, based on the data collected at the European level in the Summary Notification Information Format database. During this time period, a total of 1687 field trial notifications were received. The number of field trial notifications dropped by 76% between 1998 and 2001, mainly due to the de facto moratorium in place since 1999. Input traits (77%) dominated the field trial notifications during the last decade, while output traits were relevant in only 18% of all notifications, with a decreasing relevance during the last six years. In particular, field trial notifications on molecular farming were almost absent in the EU. Large companies focused their field trials on crops with a high grown area in the European Union and resistance traits, while public institutions showed interest in a large diversity of plants and traits. Finally, some conclusions on future impacts of the results of the study are drawn in this article. Publication Types: Review PMID: 15612507 [PubMed - indexed for MEDLINE] 1113: Environ Biosafety Res. 2004 Apr-Jun;3(2):71-2. Another look at food aid in Africa. Koch M. Publication Types: Editorial PMID: 15612503 [PubMed - indexed for MEDLINE] 1114: Wei Sheng Yan Jiu. 2004 Sep;33(5):575-8. [Subchronic toxicity test of transgenic rice] [Article in Chinese] Li Y, Piao J, Zhuo Q, Chen X, Chen X, Yang X. Institute of Nutrition and Food Safety, Chinese Center for Disease Control and Prevention, Beijing 100050, China. OBJECTIVE: To observe the subchronic toxic effects of transgenic rice on rats. METHODS: Based on gender and weight, 144 Wistar rats were randomly divided into three groups: transgenic rice group, nontransgenic rice group and AIN93G normal control group. They were fed for 90 days. Rats in transgenic rice group were fed with 73.2% transgenic rice. Rats in non-transgenic rice group were fed with 73.7% non-transgenic rice. Indicators were the following: body weight, body length, blood routine test, blood biochemistry test, organ weight, bone density and organ pathological examination. RESULTS: Compared with non-transgenic rice group, glucose was lower while cholesterol and HDLD concentration were higher in transgenic rice group in mid-experiment. But all these differences disappeared at the end of experiment. ALT activity in transgenic rice were higher than that in non-transgenic rice group. Compared with AIN93G normal control group, body weight, TG, HDLD were higher and glucose was lower in transgenic rice group in mid-experiment and all these differences disappeared at the end of the experiment. No abnormality was found with the pathological examination on brain, heart, spleen, lung, kidney, stomach, duodenum, adrenal gland, spermary and ovary. CONCLUSION: There were not enough evidences to confirm that transgenic rice had adverse effects on the rat. Publication Types: English Abstract Research Support, Non-U.S. Gov't PMID: 15612484 [PubMed - indexed for MEDLINE] 1115: Wei Sheng Yan Jiu. 2004 Sep;33(5):565-9. [Safety assessment of GM yeast feed additive with cecropin CAD gene] [Article in Chinese] Deng P, Fang S, Yang D, Jiang L, Yu X, Huang Y, Huang Z. Shenzhen Center for Disease Control and Prevention, Shenzhen 518020, China. OBJECTIVE: To evaluate the safety of GM yeast feed additive with cecropin CAD and to study and set up a model of Safety assessment for GM feed and detecting method. METHODS: To ensure the safety of the GM products, it has been done that to detect and value the safety of receptor organisms and expression products of extrinsic gene, the genetic stability of biologic properties of genomic modified yeast feed and condition of transfer and cumulation of anti-bacterial peptide and its products in circumstance and the feeded animals. RESULT AND CONCLUSION: The receptor animals and expression products of extrinic gene are safe, and the genomic modified products have steady genetic characters. The cectopin CAD neither cumulates in feeded animal nor releases into environment. The genomic modified feed additive is safe. Publication Types: English Abstract Research Support, Non-U.S. Gov't PMID: 15612481 [PubMed - indexed for MEDLINE] 1116: Environ Biosafety Res. 2003 Jul-Sep;2(3):181-206. Effects of Bacillus thuringiensis on non-target herbivore and natural enemy assemblages in tropical irrigated rice. Schoenly KG, Cohen MB, Barrion AT, Zhang W, Gaolach B, Viajante VD. Department of Biological Sciences, California State University, Stanislaus, Turlock, CA 95382, USA. kgschoenly@science.csustan.edu Endotoxins from Bacillus thuringiensis (Bt) produced in transgenic pest-resistant Bt crops are generally not toxic to predatory and parasitic arthropods. However, elimination of Bt-susceptible prey and hosts in Bt crops could reduce predator and parasitoid abundance and thereby disrupt biological control of other herbivorous pests. Here we report results of a field study evaluating the effects of Bt sprays on non-target terrestrial herbivore and natural enemy assemblages from three rice (Oryza sativa L.) fields on Luzon Island, Philippines. Because of restrictions on field-testing of transgenic rice, Bt sprays were used to remove foliage-feeding lepidopteran larvae that would be targeted by Bt rice. Data from a 546-taxa Philippines-wide food web, matched abundance plots, species accumulation curves, time-series analysis, and ecostatistical tests for species richness and ranked abundance were used to compare different subsets of non-target herbivores, predators, and parasitoids in Bt sprayed and water-sprayed (control) plots. For whole communities of terrestrial predators and parasitoids, Bt sprays altered parasitoid richness in 3 of 3 sites and predator richness in 1 of 3 sites, as measured by rarefaction (in half of these cases, richness was greater in Bt plots), while Spearman tests on ranked abundances showed that correlations, although significantly positive between all treatment pairs, were stronger for predators than for parasitoids, suggesting that parasitoid complexes may have been more sensitive than predators to the effects of Bt sprays. Species accumulation curves and time-series analyses of population trends revealed no evidence that Bt sprays altered the overall buildup of predator or parasitoid communities or population trajectories of non-target herbivores (planthoppers and leafhoppers) nor was evidence found for bottom-up effects in total abundances of non-target species identified in the food web from the addition of spores in the Bt spray formulation. When the same methods were applied to natural enemies (predators and parasitoids) of foliage-feeding lepidopteran and non-lepidopteran (homopteran, hemipteran and dipteran) herbivores, significant differences between treatments were detected in 7 of 12 cases. However, no treatment differences were found in mean abundances of these natural enemies, either in time-series plots or in total (seasonal) abundance. Analysis of guild-level trajectories revealed population behavior and treatment differences that could not be predicted in whole-community studies of predators and parasitoids. A more conclusive test of the impact of Bt rice will require field experiments with transgenic plants, conducted in a range of Asian environments, and over multiple cropping seasons. Publication Types: Comparative Study Research Support, U.S. Gov't, Non-P.H.S. PMID: 15612416 [PubMed - indexed for MEDLINE] 1117: Environ Biosafety Res. 2003 Jul-Sep;2(3):161-71. Making the EU "risk window" transparent: the normative foundations of the environmental risk assessment of GMOs. Jensen KK, Gamborg C, Madsen KH, Jørgensen RB, von Krauss MK, Folker AP, Sandøe P. Centre for Bioethics and Risk Assessment, Department of Education, Philosophy and Rhetoric, University of Copenhagen, Njalsgade 80, 2300 Copenhagen S, Denmark. kkjensen@hum.ku.dk In Europe, there seems to be widespread, morally based scepticism about the use of GMOs in food production. In response to this scepticism, the revised EU directive 2001/18/EC on the deliberate release into the environment of genetically modified organisms stresses the importance of respecting ethical principles recognized in the Member States. However, the directive fails to reflect the critical role of value judgements in scientific risk assessment and any subsequent approval procedure. In this paper we argue that it is important to make all ethically relevant assumptions involved in the approval procedure transparent and thus available for public scrutiny. Mapping the value judgements that are made in an environmental risk assessment and approval procedure, we describe the political liberal nature of the EU legislation. We then look more closely at the prescriptions for environmental risk assessment and approval of GMOs outlined in the directive. An environmental risk assessment views the world through a "risk window" that only makes visible that which has been predefined as a relevant risk. The importance of the value judgements that define the risk window consists in limiting the information the risk assessment can provide. In the penultimate section of the paper, the significance of the risk window is demonstrated through a case study of the approval of glyphosate resistant fodder beets (Beta vulgaris L. ssp. vulgaris) in Denmark. PMID: 15612414 [PubMed - indexed for MEDLINE] 1118: Environ Biosafety Res. 2003 Oct-Dec;2(4):263-76. Transgenic Bt-producing Brassica napus: Plutella xylostella selection pressure and fitness of weedy relatives. Mason P, Braun L, Warwick SI, Zhu B, Stewart CN Jr. Agriculture and Agri-Food Canada, Eastern Cereal and Oilseed Research Centre, K. W. Neatby Bldg., Ottawa, Ontario, K1A 0C6, Canada. masonp@agr.gc.ca Release of transgenic insect-resistant crops creates the potential not only for the insect pest to evolve resistance but for the escape of transgenes that may confer novel or enhanced fitness-related traits through hybridization with their wild relatives. The differential response of diamondback moth (Plutella xylostella) populations in eastern and western Canada to Bt-producing (GT) Brassica napus and the potential for enhanced fitness of GT B. napus and weedy GT Brassica rapa x B. napus hybrid populations (F1, BC1, BC2) were studied. Comparative bioassays using neonates and 4th instars showed that GT B. napus and GT B. rapa x B. napus hybrids are lethal to larvae from both populations. No measurable plant fitness advantage (reproductive dry weight) was observed for GT B. napus (crop) and GT B. rapa x B. napus hybrid populations at low insect pressure (1 larva per leaf). At high insect densities (>10 larvae per leaf), vegetative plant weight was not significantly different for GT B. napus and non-GT B. napus, whereas reproductive plant weight and proportion of reproductive material were significantly higher in GT B. napus. Establishment of the Bt trait in wild B. rapa populations may also increase its competitive advantage under high insect pressure. Publication Types: Research Support, Non-U.S. Gov't PMID: 15612282 [PubMed - indexed for MEDLINE] 1119: Environ Mol Mutagen. 2005;45(1):70-9. 17 Beta-estradiol and not genistein modulates lacI mutant frequency and types of mutation induced in the heart of ovariectomized big blue rats treated with 7, 12-dimethylbenz[a]anthracene. Manjanatha MG, Shelton SD, Rhodes BS, Bishop ME, Lyn-Cook LE, Aidoo A. Division of Genetic and Reproductive Toxicology, Food and Drug Administration/National Center for Toxicological Research, Jefferson, Arkansas 72079, USA. mmanjanatha@nctr.fda.gov In industrialized countries, heart disease rates are higher among women after menopause. Recent studies indicate that consumption of phytoestorogens, e.g., isoflavones such as genistein (GE), may have potential cardiovascular health benefits; however, no studies have evaluated the effect of these agents on toxicant-induced damage in the heart. Since estrogen receptors are found in the heart, and GE mimics estrogenic effects, we have examined whether or not dietary GE or 17 beta-estradiol (E2) modulates the lacI mutant frequency (MF) in the heart of ovariectomized (OVX) Big Blue rats exposed to the model carcinogen 7,12-dimethylbenz[a]anthracene (DMBA). Groups of female rats were administered 80 mg/kg DMBA or vehicle by gavage and were chronically fed with diets containing 0, 250, or 1,000 microg/g GE or 5 microg/g E2. Sixteen weeks after carcinogen treatment, the animals were sacrificed and the hearts were removed and processed for determining the frequency and types of mutations in the heart tissue. GE and E2 supplementation alone resulted in nonsignificant increases in MF. The DMBA-induced lacI MF in the heart was sevenfold higher than the control (119.8 +/- 18.7 x 10(-6) vs. 17.4 +/- 3.2 x 10(-6); P < 0.001). GE in the diet had no significant effect on DMBA mutagenicity, while feeding E2 to DMBA-treated rats caused a significant reduction in the MF (119.8+/- 18.7 x 10(-6) vs. 61.4 +/- 13.5 x 10(-6); P < 0.017). DNA sequence analysis revealed that the majority of DMBA-induced mutations in rats fed control diet were A:T-->T:A (42%) and G:C-->T:A (19%) transversions, followed by G:C-->A:T (13%) and A:T-->G:C (8%) transitions. Feeding E2 altered the DMBA-induced mutational spectra by decreasing A:T-->T:A (23%) and G:C-->T:A (13%) transversions and increasing G:C-->A:T (24%) and A:T-->G:C (21%) transitions. Taken together, the results suggest that DMBA can induce gene mutations in heart tissue of OVX rats, and while dietary GE had little or no effect on DMBA-induced mutation, dietary E2 reduced the mutagenicity of DMBA. 2004 Wiley-Liss, Inc. PMID: 15611980 [PubMed - indexed for MEDLINE] 1120: Biotechnol Adv. 2005 Jan;23(1):81-5. Epub 2004 Oct 27. The thioredoxin h system: potential applications. Joudrier P, Gautier MF, de Lamotte F, Kobrehel K. UMR PIA (CIRAD-INRA-ENSA), 2, Place Viala, 34060 Montpellier cedex, France. joudrier@ensam.inra.fr The thioredoxin h system has the specific capability to reduce intramolecular disulfide bonds of proteins, thereby modifying their tertiary structure. It is involved in many processes: in the activation or deactivation of enzymes and enzyme inhibitors and in the germination process. This system can be used to improve the breadmaking quality of wheat by strengthening the dough. It can also decrease the epitope accessibility, then modifying the response of the IgE immune system. Transgenic barley and wheat have been created to confirm the functionality of the NADP-dependent thioredoxin h system. Publication Types: Review PMID: 15610969 [PubMed - indexed for MEDLINE] 1121: J Appl Microbiol. 2005;98(1):127-35. Development of food-grade cloning and expression vectors for Lactococcus lactis. Liu CQ, Su P, Khunajakr N, Deng YM, Sumual S, Kim WS, Tandianus JE, Dunn NW. Department of Biotechnology, University of New South Wales, Sydney, Australia. chun.liu@dsto.defence.gov.au AIMS: To develop food-grade cloning and expression vectors for use in genetic modification of Lactococcus lactis. METHODS AND RESULTS: Two plasmid replicons and three dominant selection markers were isolated from L. lactis and used to construct five food-grade cloning vectors. These vectors were composed of DNA only from L. lactis and contained no antibiotic resistance markers. Three of the vectors (pND632, pND648 and pND969) were based on the same plasmid replicon and carried, either alone or in combination, the three different selectable markers encoding resistance to nisin, cadmium and/or copper. The other two (pND965DJ and pND965RS) were derived from a cadmium resistance plasmid, and carried a constitutive promoter and a copper-inducible promoter, respectively, immediately upstream of a multicloning site. All vectors were stable in L. lactis LM0230 for at least 40 generations without selection pressure. The two groups of vectors were compatible in L. lactis LM0230. The vectors pND648 and pND965RS, as representatives of the two groups, were transferred successfully by electroporation into and maintained in an industrial strain of L. lactis. The usefulness of the vectors was further demonstrated by expressing a phage resistance gene (abiI) in another industrial strain of L. lactis. CONCLUSIONS: The five food-grade vectors constructed are potentially useful for industrial strains of L. lactis. SIGNIFICANCE AND IMPACT OF THE STUDY: These vectors represent a new set of molecular tools useful for food-grade modifications of L. lactis. PMID: 15610425 [PubMed - indexed for MEDLINE] 1122: J Appl Microbiol. 2005;98(1):43-55. Phenylacetic acid-producing Rhizoctonia solani represses the biosynthesis of nematicidal compounds in vitro and influences biocontrol of Meloidogyne incognita in tomato by Pseudomonas fluorescens strain CHA0 and its GM derivatives. Siddiqui IA, Shaukat SS. Soil Biology and Ecology Laboratory, Department of Botany, University of Karachi, Karachi, Pakistan. imran_7585@yahoo.com AIMS: The aim of the present investigation was to determine the influence of Rhizoctonia solani and its pathogenicity factor on the production of nematicidal agent(s) by Pseudomonas fluorescens strain CHA0 and its GM derivatives in vitro and nematode biocontrol potential by bacterial inoculants in tomato. METHODS AND RESULTS: One (Rs7) of the nine R. solani isolates from infected tomato roots inhibited seedling emergence and caused root rot in tomato. Thin layer chromatography revealed that culture filtrates of two isolates (Rs3 and Rs7) produced brown spots at Rf-values closely similar to synthetic phenylacetic acid (PAA), a phytotoxic factor. Filtrates from isolate Rs7, amended with the growth medium of P. fluorescens, markedly repressed nematicidal activity and PhlA'-'LacZ reporter gene expression of the bacteria in vitro. On the contrary, isolate Rs4 enhanced nematicidal potential of a 2,4-diacetylphloroglucinol overproducing mutant, CHA0/pME3424, of P. fluorescens strain CHA0 in vitro. Therefore, R. solani isolates Rs4 and Rs7 were tested more rigorously for their potential to influence biocontrol effectiveness of the bacterial agents. Methanol extract of the culture filtrates of PAA-producing isolate Rs7 resulting from medium amended with phenylalanine enhanced fungal repression of the production of nematicidal agents by bacteria, while amendments with zinc or molybdenum eliminated such fungal repression, thereby restoring bacterial potential to cause nematode mortality in vitro. A pot experiment was carried out, 3-week-old tomato seedlings were infested with R. solani isolates Rs4 or Rs7 and/or inoculated with Meloidogyne incognita, the root-knot nematode. The infested soil was treated with aqueous cell suspensions (10(8) CFU) of P. fluorescens strain CHA0 or its GM derivatives or left untreated (as a control). Observations taken 45 days after nematode inoculation revealed that, irrespective of the bacterial treatments, galling intensity per gram of fresh tomato roots was markedly higher in soil amended with isolate Rs4 than in Rs7-amended soils. Soil amendments with R. solani and the bacterial antagonists resulted in substantial reductions of the number of galls per gram of root. These results are contradictory to those obtained under in vitro conditions where culture filtrates of PAA-positive Rs7 repressed the production of nematicidal compounds. Plants grown in Rs7-amended soils, with or without bacterial inoculants, had lesser shoot and root weights than plants grown in nonamended or Rs4-amended soils. Moreover, amendments with Rs7 substantially retarded root growth and produced necrotic lesions that reduced the number of entry sites for invasion and subsequent infection by nematodes. Populations of P. fluorescens in the tomato rhizosphere were markedly higher in Rs7-amended soils. CONCLUSIONS: PAA-producing virulent R. solani drastically affects the potential of P. fluorescens to cause death of M. incognita juveniles in vitro and influences bacterial effectiveness to suppress nematodes in tomato roots. SIGNIFICANCE AND IMPACT OF THE STUDY: As most agricultural soils are infested with root-infecting fungi, including R. solani, it is likely that some PAA-producing isolates of the fungus may also be isolated from such soils. The inhibitory effect of PAA-producing R. solani on the biosynthesis of nematicidal agent(s) critical in biocontrol may reduce or even eliminate the effectiveness of fluorescent pseudomonads against root-knot nematodes, both in nursery beds and in field conditions. Introduction of bacterial inoculants, for the control of any plant pathogen, should be avoided in soils infested with PAA-producing R. solani. Alternatively, the agents could be applied together with an appropriate quantity of fungicide or chemicals such as zinc to create an environment more favourable for bacterial biocontrol action. PMID: 15610416 [PubMed - indexed for MEDLINE] 1123: Appetite. 2005 Feb;44(1):115-22. Food and values: an examination of values underlying attitudes toward genetically modified- and organically grown food products. Dreezens E, Martijn C, Tenbült P, Kok G, de Vries NK. Department of Experimental Psychology, Faculty of Psychology, Maastricht University, P.O. Box 616, 6200 MD Maastricht, The Netherlands. e.dreezens@psychology.unimaas.nl This study addresses which specific values play a role in predicting participants' attitudes toward genetically modified food (GMF) and organically grown food (OGF). The first central question is whether the attitudes towards GMF and OGF are influenced by specific values and beliefs. The second central question is whether the attitudes towards GMF and OGF are related to each other, and whether the specific values underlying these two attitudes are also related to each other. A total of 100 participants responded to the Schwartz Value Survey and two questionnaires about GMF and organically grown food. When respondents scored high on the value power (dominance, submission), they rated GMF positively and OGF more negatively. Respondents who rated the value universalism (welfare for all people and protection of nature) high, rated OGF as positive. Furthermore, the relationship between attitudes and values was mediated by beliefs. These findings imply a meaningful relationship between specific values, beliefs, and these food-related attitudes, and suggest that values might play a role in explaining attitudes toward GMF and OGF products. Publication Types: Research Support, Non-U.S. Gov't PMID: 15604038 [PubMed - indexed for MEDLINE] 1124: Nature. 2004 Dec 16;432(7019):799. Comment on: Nature. 2004 Oct 21;431(7011):883. Media affect opinions less than they would like. Melchett P. Publication Types: Comment Letter PMID: 15602522 [PubMed - indexed for MEDLINE] 1125: Zhi Wu Sheng Li Yu Fen Zi Sheng Wu Xue Xue Bao. 2004 Apr;30(2):121-6. [Excision of selectable marker gene from transgenic plant] [Article in Chinese] Lu HJ, Gong ZX. Key Laboratory of Proteomics, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China. gongzx@sumn.shcnc.ac.cn Currently used plant transformation systems require selectable marker genes encoding antibiotic or herbicide resistance, along with the gene of interest, to select transformed cells from a large population of mostly untransformed cells. The continued presence of these selectable markers, especially in food crop, is of increasing public concern. The generation of selectable marker-free transgenic plant is one of the new projects in plant biotechnology research. Two techniques, segregation excision and recombination excision, for removal of selectable marker genes are described in this article. The advances in producing selectable marker-free transgenic plants are reviewed too. Publication Types: English Abstract Research Support, Non-U.S. Gov't Review PMID: 15599035 [PubMed - indexed for MEDLINE] 1126: Pest Manag Sci. 2005 Mar;61(3):292-300. Risks and consequences of gene flow from herbicide-resistant crops: canola (Brassica napus L) as a case study. Légère A. Soils and Crops Research and Development Centre, Agriculture and Agri-Food Canada, 2560 boul Hochelaga, Sainte-Foy, QC, G1V 2J3, Canada. legerea@agr.gc.ca Data from the literature and recent experiments with herbicide-resistant (HR) canola (Brassica napus L) repeatedly confirm that genes and transgenes will flow and hybrids will form if certain conditions are met. These include sympatry with a compatible relative (weedy, wild or crop), synchrony of flowering, successful fertilization and viable offspring. The chance of these events occurring is real; however, it is generally low and varies with species and circumstances. Plants of the same species (non-transgenic or with a different HR transgene) in neighbouring fields may inherit the new HR gene, potentially generating plants with single and multiple HR. For canola, seed losses at harvest and secondary dormancy ensures the persistence over time of the HR trait(s) in the seed bank, and the potential presence of crop volunteers in subsequent crops. Although canola has many wild/weedy relatives, the risk of gene flow is quite low for most of these species, except with Brassica rapa L. Introgression of genes and transgenes in B rapa populations occurs with apparently little or no fitness costs. Consequences of HR canola gene flow for the agro-ecosystem include contamination of seed lots, potentially more complex and costly control strategy, and limitations in cropping system design. Consequences for non-agricultural habitats may be minor but appear largely undocumented. Minister of Public Works and Government Services Canada 2005 Publication Types: Review PMID: 15593291 [PubMed - indexed for MEDLINE] 1127: Biochimie. 2004 Nov;86(11):793-8. Enzymes for transgenic biosynthesis of long-chain polyunsaturated fatty acids. Huang YS, Pereira SL, Leonard AE. Strategic Research, Ross Products Division, Abbott Laboratories, 625 Cleveland Avenue, Columbus, OH 43215, USA. vic.huang@abbott.com Polyunsaturated fatty acids (PUFAs) are important for the normal development and function of all organisms, and are essential in maintaining human health. Impaired PUFA metabolism is thought to be associated with pathogenesis of many chronic diseases. Dietary supplementation of PUFAs, such as gamma-linolenic acid, arachidonic acid, eicosapentaenoic acid, and docosahexaenoic acid, which bypass the defective or dysfunctional steps of the biosynthetic pathway has been found to significantly alleviate the symptoms of the disease. These findings have drawn a great deal of interest from general public and food manufacturers. As the demand of these beneficial PUFAs has drastically increased in recent years, there are also increasing efforts in finding the alternate sources of PUFAs that are more economical and sustainable. One option is to modify the oil-seed crops to produce PUFAs through genetic engineering technique. This review examines the isolation, identification and expression of genes encoding the enzymes required for the biosynthesis of the above mentioned PUFAs in plants. Publication Types: Review PMID: 15589688 [PubMed - indexed for MEDLINE] 1128: Toxicology. 2005 Jan 15;206(2):195-205. Evaluation of an in vitro method for the measurement of specific IgE antibody responses: the rat basophilic leukemia (RBL) cell assay. Dearman RJ, Skinner RA, Deakin N, Shaw D, Kimber I. Syngenta Central Toxicology Laboratory, Alderley Park, Macclesfield, Cheshire SK10 4TJ, UK. rebecca.dearman@syngenta.com The evaluation of allergenic potential is a key parameter in the safety assessment of novel proteins, including those expressed in genetically modified crops and foodstuffs. The majority of allergic reactions to food proteins are immediate type hypersensitivity reactions in which the principal biological effector is IgE antibody; the accurate measurement of specific IgE antibody is therefore a critical factor in experimental systems designed to characterize protein allergenic potential. Due to the presence of much higher concentrations of other immunoglobulin isotypes, the assessment of specific serum IgE antibody poses substantial technical challenges. We have examined the utility of the rat basophilic leukemia (RBL) cell line for the measurement of murine IgE responses. RBL cells were sensitized with mouse monoclonal anti-dinitrophenyl (DNP) IgE antibody and challenged with DNP-albumin conjugates with various hapten substitution ratios (SR). Polyclonal anti-OVA IgE antisera were also assessed for activity in the RBL assay. Results were compared with titers measured in homologous passive cutaneous anaphylaxis (PCA) assay. Marked degranulation of RBL cells was induced by conjugates with SRs of between 16 and 32, whereas conjugates with lower SRs (of 10 or 3) failed to elicit significant serotonin release. All conjugates were able to induce mast cell degranulation in vivo in a PCA assay. Anti-OVA antisera with PCA titers of 1/32 to 1/64 failed to stimulate RBL cell degranulation, whereas high titer antibody (1/2048 to 1/4096 by PCA) induced a positive RBL cell response. Successful stimulation of RBL cell degranulation requires not only appropriate epitope densities but also high affinity antibody. These data indicate that this assay is inappropriate for the routine analysis of specific polyclonal IgE antibody responses such as those that are induced by exposure to complex protein allergens. Publication Types: Comparative Study Evaluation Studies PMID: 15588913 [PubMed - indexed for MEDLINE] 1129: J Polit Philos. 2000 Jun;8(2):154-75. Rights to life? On nature, property and biotechnology. Meyer JM. Goverment and Politics, Humboldt State University, USA. PMID: 15586933 [PubMed - indexed for MEDLINE] 1130: Sci Eng Ethics. 2004 Oct;10(4):705-16. How Japanese students reason about agricultural biotechnology. Maekawa F, Macer D. Institute of Biological Sciences, University of Tsukuba, Japan. Many have claimed that education of the ethical issues raised by biotechnology is essential in universities, but there is little knowledge of its effectiveness. The focus of this paper is to investigate how university students assess the information given in class to make their own value judgments and decisions relating to issues of agricultural biotechnology, especially over genetically modified organisms (GMOs). Analysis of homework reports related with agricultural biotechnology after identification of key concepts and ideas in each student report is presented. The ideas were sorted into different categories. The ideas were compared with those in the reading materials using the same categories. These categories included: concern about affects on humans, affects on the environment, developing countries and starvation, trust in industry, responsibility of scientists, risk perception, media influence, need for (international) organizations or third parties, and information dissemination. What was consistent through the different years was that more than half of the students took a "neutral" position. A report was scored as "neutral" when the report included both the positive and negative side of an issue, or when the student could not make a definite decision about the use of GMOs and GM food. While it may be more difficult to defend a strong ''for" or "against" position, some students used logical arguments successfully in doing so. Sample comments are presented to depict how Japanese students see agricultural technology, and how they value its application, with comparisons to the general social attitudes towards biotechnology. PMID: 15586729 [PubMed - indexed for MEDLINE] 1131: Arh Hig Rada Toksikol. 2004 Nov;55(4):301-12. [Genetically modified organisms in food--production, detection and risks] [Article in Croatian] Zeljezić D. Institut za medicinska istrazivanja i medicinu rada, Zagreb. dzeljezi@imi.hr The first genetically modified plant (GMP) was a tobacco resistant to antibiotics in 1983. In 1996, the first genetically altered crop, a delayed-ripening tomato was commercially released. In the year 2003, the estimated global area of GM crops for was 67.7 million hectares. To produce such a plant a gene of interest has to be isolated from the donor. Together with a promoter, terminator sequence and marker gene it has to be introduced into the plant cell which is then stimulated to generate a whole GMP expressing new characteristics (herbicide/insect resistance, delayed ripening). The last few months have seen a strong public debate over genetically modified organisms which has raised scientific, economic, political, and ethical issues. Some questions concerning the safety of GMPs are still to be answered, and decisions about their future should be based on scientifically validated information. Publication Types: English Abstract Review PMID: 15584557 [PubMed - indexed for MEDLINE] 1132: Med Ethics. 2001 Fall:6-7. Reason and repugnance. Callahan D, Magnus D. The Hastings Center, Garrison, NY, USA. PMID: 15584188 [PubMed - indexed for MEDLINE] 1133: Nat Biotechnol. 2004 Dec;22(12):1501; discussion 1501. Comment on: Nat Biotechnol. 2004 Sep;22(9):1055. No Munich on GM crops. Malvoisin P, Grausz JD. Publication Types: Comment Letter PMID: 15583648 [PubMed - indexed for MEDLINE] 1134: Plant Cell Physiol. 2004 Nov;45(11):1586-94. Two distinct redox signaling pathways for cytosolic APX induction under photooxidative stress. Yabuta Y, Maruta T, Yoshimura K, Ishikawa T, Shigeoka S. Department of Food and Nutrition, Faculty of Agriculture, Kinki University, 3327-204 Nakamachi, Nara, 631-8505 Japan. The cross-talk of two redox factors, H2O2 accumulation and redox status of photosynthetic electron transport (PET) in tobacco chloroplasts, on the expression of cytosolic ascorbate peroxidase (cAPX) was studied. Transgenic tobacco plants, which expressed respectively the Escherichia coli catalase and spinach thylakoid-membrane-bound APX in chloroplasts and showed increased tolerance to photooxidative stress, were used for evaluation of the relationship between H2O2 accumulation or change of redox status of PET and cAPX induction under photooxidative stress condition. There was no difference in the increase in the transcript level of cAPX in the first 1 h after irradiation with high-intensity light between the wild-type and either of the transgenic plants. The transcript level of cAPX in the wild-type showed a steady gradual increase during the next 5 h, while that in the transgenic plants during this period was stable. The H2O2 level of wild-type plants increased after 1 h, while those of transgenic plants remained constant. The decrease in q(p) value in all types of plants occurred earlier than the increase in H2O2 level. These results indicate that the induction of cAPX expression is caused by a redox change in PET, probably through the plastquinone pool at an early stage and thereafter by an increase in the cellular H2O2 level. Publication Types: Research Support, Non-U.S. Gov't PMID: 15574834 [PubMed - indexed for MEDLINE] 1135: J Nutr. 2004 Dec;134(12):3264-9. Metabolic adaptations of three inbred strains of mice (C57BL/6, DBA/2, and 129T2) in response to a high-fat diet. Funkat A, Massa CM, Jovanovska V, Proietto J, Andrikopoulos S. The University of Melbourne, Department of Medicine, (AH/NH), Heidelberg Repatriation Hospital, Heidelberg Heights, Victoria 3081 Australia. Although it is now becoming more evident that the strain of mouse used to generate genetically modified models for the study of endocrine disorders contributes to the ensuing phenotype, metabolic characterization of these common strains used to produce genetically altered mice has been limited. The aim of this study therefore was to measure various metabolic parameters in C57BL/6, DBA/2, and 129T2 mice fed a control or a high-fat diet. Mice were fed either a control (7 g/100 g) or a high-fat (60 g/100 g) diet for 6 wk. During wk 6, spontaneous and voluntary physical activity and resting energy expenditure were determined. DBA/2 mice that consumed the control diet gained more weight and had larger regional fat pad depots than either C57BL/6 or 129T2 mice (P < 0.05). Spontaneous and voluntary activity was lower in 129T2 mice compared with DBA/2 or C57BL/6 mice (P < 0.05). Resting energy expenditure (corrected for body weight) was greater in C57BL/6 mice than in DBA/2 or 129T2 mice (P < 0.05), whereas glucose and fat oxidation did not differ among the 3 strains of mice. Plasma glucose concentrations in food-deprived mice were higher and insulin concentrations lower in 129T2 compared with C57BL/6 mice (P < 0.05), but were not affected by the high-fat diet in any of the 3 strains tested. This study shows that these 3 commonly used inbred strains of mice have different inherent metabolic characteristics. It further highlights that the background strain used to produce genetically modified mice is critical to the resultant phenotype. Publication Types: Comparative Study Research Support, Non-U.S. Gov't PMID: 15570023 [PubMed - indexed for MEDLINE] 1136: Shokuhin Eiseigaku Zasshi. 2004 Aug;45(4):207-11. [Examination of DNA extract from kernels and processed foods using silica-base resin] [Article in Japanese] Nakama A, Morishita F. Nutrition College, Osaka Institute of Public Health and Environmental Sciences: 8-34, Tojo-cho, Tennoji-ku, Osaka 543-0026, Japan. A rapid and simple DNA extraction method is needed to detect genetically modified recombinant DNA in soybean kernels and processed foods. However, since various kernels and processed foods differ greatly in form, a uniform DNA extraction method has proved elusive. The silica-base resin DNA extraction method does not use any organic solvent, and the operation is simple and the cost per extraction is low, although the frequency of its use is very low and few domestic reports exist. We therefore studied suitable conditions for a silica-base resin method. We also developed the method to get more pure DNA from soybean kernels. The silica-base resin method was found to be adequate for extracting DNA from various processed foods for PCR amplification with endogenous gene primers. In the case of DNA extraction from soybean kernels, pure DNA could be efficiently extracted after pre-heating the soybean suspension in TNE buffer. The extracted DNA showed higher ratios of absorption at 260 nm/280 nm and 260 nm/230 nm than those for samples obtained with previous methods. Moreover, our observations suggested that the extraction time could be reduced to within 30 min for processed foods such as tofu. Publication Types: English Abstract PMID: 15568472 [PubMed - indexed for MEDLINE] 1137: Shokuhin Eiseigaku Zasshi. 2004 Aug;45(4):184-90. [Detection of genetically modified organisms obtained from food samples ] [Article in Japanese] Monma K, Araki R, Ichikawa H, Sato M, Uno N, Sato K, Tobe T, Kuribara H, Matsuoka T, Hino A, Saito K. Tokyo Metropolitan Institute of Public Health: 3-24-1, Hyakunin-cho, Shinjuku-ku, Tokyo 169-0073, Japan. Genetially modified organisms (GMOs) were explored in food samples obtained from November 2000 to March 2003 in the Tokyo area by using PCR and real-time PCR techniques. The existence of Roundup Ready Soybean (RRS) was surveyed in processed foods derived from soybeans, such as tofu, boiled soybean, kinako, nama-age, abura-age, natto, miso, soymilk and yuba. RRS was detected in 3 of 37 tofu, 2 of 3 nama-age, 2 of 3 yuba and 3 of 3 abura-age samples. The CBH351 in 70 processed corn foods, NewLeaf Plus and NewLeaf Y in 50 processed potato foods, and 55-1 papaya in 16 papayas were surveyed. These GMOs were not detected among the samples. Qualitative and quantitative analyses of RRS and genetically modified (GM) corn were performed in soybean, corn and semi-processed corn products such as corn meal, corn flour and corn grits. RRS was detected in 42 of 178 soybean samples, and the amount of RRS in RRS-positive samples was determined. The content was in the range of 0.1-1.4% in identity-preserved soybeans (non-GMO), and 49.8-78.8% in non-segregated soybeans. On the other hand, GM corns were detected in 8 of 26 samples. The amount of GM corn in GM corn-positive samples was in the range of 0.1-2.0%. Publication Types: English Abstract PMID: 15568468 [PubMed - indexed for MEDLINE] 1138: Science. 2004 Nov 26;306(5701):1458-9. Agriculture. China could be first nation to approve sale of GM rice. Lei X. Publication Types: News PMID: 15567824 [PubMed - indexed for MEDLINE] 1139: Risk Anal. 2004 Oct;24(5):1385-93. Psychological determinants of willingness to taste and purchase genetically modified food. Townsend E, Campbell S. Institute for the Study of Genetics, Biorisks and Society and Department of Philosophy, University of Nottingham, UK. Ellen.Townsend@nottingham.ac.uk Decreasing acceptance of biotechnologies over time has been reported in Europe. Studies claim that attitudes are negative, even hostile, and that people are very worried about genetic engineering in food and medicine. However, such studies are mostly based on surveys and these have significant methodological problems, such as low response rates, which may indicate that only those with strong views respond, thus biasing the sample. Here an alternative method, involving "topic-blind" recruitment of participants and a behavioral measure (food tasting), was used. We show that in a topic-blind sample of 100 individuals, 93% willingly tasted and ate what they believed to be genetically modified (GM) food in an experimental setting, and 48% said they would buy GM food in the future, results that are surprising in the context of other reports about attitudes and intentions toward GM food. Purchasers and nonpurchasers differed in their attitudes toward GM food on key risk-related scales (particularly on a dread-not dread scale--a measure of integral affect--and an ethical-unethical scale). Despite these differences, however, and despite their negative attitude, most nonpurchasers (85.7%) still tasted the GM apple. Incidental affect (state stress and trait worry) was not found to influence risk-related judgments about GM food. Integral affect (dread of GM plants and animals used for food) and concerns about the future risks of GM animals in food were found to be key predictors of willingness to purchase GM food. Publication Types: Research Support, Non-U.S. Gov't PMID: 15563302 [PubMed - indexed for MEDLINE] 1140: Risk Anal. 2004 Oct;24(5):1369-84. Effects of context and feelings on perceptions of genetically modified food. Townsend E, Clarke DD, Travis B. Institute of Genetics, Biorisks and Society and School of Psychology, University of Nottingham, UK. Ellen.Townsend@nottingham.ac.uk Recently, there has been a surge of interest in the role of feelings in framing perceptions and decisions about risk, yet no study has specifically examined the impact of feelings on perceptions/judgments about biotechnology. This exploratory study investigated current perceptions of genetically modified (GM) food to examine (1) the effects of context (making judgments about GM food at the same time as rating other current areas of concern), and (2) the effect of feelings of dread (integral affect) and background feelings of stress (negative incidental affect) on risk judgments about GM food. An established psychometric method (semantic differential task) used with a sample of 126 adults (recruited "topic-blind," mostly from a student population) showed that, when rated in the context of other current concerns such as human cloning and Creutzfeldt-Jakob disease (CJD), there was less concern about GM food than might have been anticipated. Participants were recruited "topic-blind" in order to ensure that they were unaware that the focus of the research was on GM food specifically (and thus preventing biased recruitment to the study). Relative to 19 other current concerns GM food was "not dreaded," not viewed as "unethical," was judged as "controllable," and was seen as the least "risky" of all the issues studied. GM food was viewed as a "hot topic," a new risk, and as relatively unnatural (although it was not the highest rated concern on this scale). Ratings of risks across concerns by individuals experiencing high levels of negative incidental affect (stress) did not differ significantly from those reporting low stress. Publication Types: Comparative Study Research Support, Non-U.S. Gov't PMID: 15563301 [PubMed - indexed for MEDLINE] 1141: Risk Anal. 2004 Oct;24(5):1311-21. Reactions to genetically modified food crops and how perception of risks and benefits influences consumers' information gathering. Wilson C, Evans G, Leppard P, Syrette J. CSIRO, Health Sciences and Nutrition, PO Box 10041, Adelaide BC SA 5000. Carlene.Wilson@csiro.au Previous research has reported strong consumer perception that genetically modified (GM) food crops may lead to adverse outcomes in a number of different areas. This is despite the widespread promulgation of the potential benefits and opportunities ascribed to the same technology by many scientists and other experts. A computer-based information gathering and evaluation task was completed by 198 adults to assess the extent to which their initial focus on the dangers or opportunities of genetic modification, or both, could be ascribed to the manner in which they gathered information on the topic (heuristically vs. systematically). Results did not confirm the hypothesis that initial focus (risks, benefits, or both) predicted ongoing information gathering and evaluation behavior. Moreover, also contrary to prediction, most participants primarily used systematic strategies when deriving their initial position, regardless of that opinion. Participants found it difficult to achieve a balanced perspective on GM food crop, even though balanced argument, as measured by order of story selection and time spent reading, was preferred as the source of information. Perceived importance is probably the most influential variable determining information gathering about issues or events to which a level of risk is attached. PMID: 15563297 [PubMed - indexed for MEDLINE] 1142: Risk Anal. 2004 Oct;24(5):1289-99. Expert and public perception of risk from biotechnology. Savadori L, Savio S, Nicotra E, Rumiati R, Finucane M, Slovic P. Dipartimento di Scienze della Cognizione e della Formazione, University of Trento, Italy. savadori@form.unitn.it Risk perceptions of a series of biotechnology applications were examined in a public (nonexpert) sample and an expert sample. Compared with the experts, the public perceived all biotechnology applications as more risky. Both groups perceived food-related applications to be riskier than medical applications. Compared with the public, experts perceived both food and medical applications as less harmful and more useful. Experts also judged the risks posed from medical biotechnology applications as more familiar and acknowledged by people and science. Lay estimates of the risk of food applications were predicted by potential harm, potential benefits, science knowledge, and familiarity; experts' estimates were predicted only by harm and benefits. Lay estimates of the risk of medical applications were predicted by potential harm; experts' estimates were predicted by potential benefits, number and type of people exposed, and science knowledge. We discuss the implications of the results for risk communication about and management of different types of biotechnologies. Publication Types: Comparative Study PMID: 15563295 [PubMed - indexed for MEDLINE] 1143: Ann Allergy Asthma Immunol. 2004 Nov;93(5 Suppl 3):S19-25. Genetic modification of food allergens. Lehrer SB. Department of Medicine, Section of Clinical Immunology, Allergy and Rheumatology, Tulane University School of Medicine, New Orleans, Louisiana 70112, USA. sblehrer@tulane.edu OBJECTIVE: To review allergen risk evaluation for genetically modified foods and our ability to predict protein allergenicity, methods that are being used to develop foods with reduced allergenic activity, and clinical aspects relative to assessing potentially allergic patients. DATA SOURCES: Information was identified using the MEDLINE database for governmental, international, and industry organizations that have considered possible unintended health effects such as food allergy and how they can be avoided. DATA SELECTION: The author's knowledge of the field was used to select articles for inclusion in this review. RESULTS: Organizations have created a decision process that has generally been successful in avoiding development of products that cause allergic reactions. Since some proteins expressed do not have any history of human exposure, risk evaluation may be more of a challenge for them. Biotechnology has also been used to try to develop foods with reduced allergenicity, and in future years such products should yield safer foods. CONCLUSIONS: Allergy risk evaluation for known allergens and genetically modified foods appears to be reasonable and provides assurance of food safety. Allergenicity evaluation of novel proteins is a more complicated process that needs to be and will be improved as our knowledge of food allergens increases. Biotechnology can be used to produce safer and healthier foods; for example, allergenicity of some foods may be reduced through biotechnology. The role of the health care professional in assessing allergic reactions to genetically modified foods is essential and should play a greater role in the interaction of consumers, industry, and regulators. Publication Types: Review PMID: 15562870 [PubMed - indexed for MEDLINE] 1144: Science. 2004 Nov 19;306(5700):1295-7. Intellectual property. Plants and intellectual property: an international appraisal. Koo B, Nottenburg C, Pardey PG. International Food Policy Research Institute, Washington, DC 20006-1002, USA. b.koo@cgiar.org Publication Types: Research Support, Non-U.S. Gov't PMID: 15550646 [PubMed - indexed for MEDLINE] 1145: Crit Rev Food Sci Nutr. 2004;44(5):361-7. Health benefits of soy isoflavonoids and strategies for enhancement: a review. McCue P, Shetty K. Program in Molecular and Cellular Biology, University of Massachusetts, Amherst, Massachusetts 01003, USA. Soybean consumption has been linked to a reduced risk for certain cancers and diseases of old age. The health benefits associated with soybean consumption have been linked to the action of isoflavonoids, the major phenolic phytochemicals found in soybean. Isoflavonoids possess numerous biological activities that may support chemoprevention through the promotion of apoptosis in diseased cells. In this study, we discuss the current state of knowledge concerning soybean isoflavonoids, their chemopreventive actions against postmenopausal health problems, cancer, and cardiovascular disease, and also biotechnology approaches toward the enrichment of soybean for isoflavonoid content. Publication Types: Review PMID: 15540649 [PubMed - indexed for MEDLINE] 1146: Science. 2004 Nov 12;306(5699):1101. What's on the label? Krebs J. Publication Types: Editorial PMID: 15539567 [PubMed - indexed for MEDLINE] 1147: Nature. 2004 Nov 11;432(7014):222-5. Environmental biosafety and transgenic potato in a centre of diversity for this crop. Celis C, Scurrah M, Cowgill S, Chumbiauca S, Green J, Franco J, Main G, Kiezebrink D, Visser RG, Atkinson HJ. Laboratory of Plant Breeding, Wageningen University, PO Box 386, 6700 AJ, Wageningen, The Netherlands. The Nuffield Council on Bioethics suggests that introgression of genetic material into related species in centres of crop biodiversity is an insufficient justification to bar the use of genetically modified crops in the developing world. They consider that a precautionary approach to forgo the possible benefits invokes the fallacy of thinking that doing nothing is itself without risk to the poor. Here we report findings relevant to this and other aspects of environmental biosafety for genetically modified potato in its main centre of biodiversity, the central Andes. We studied genetically modified potato clones that provide resistance to nematodes, principal pests of Andean potato crops. We show that there is no harm to many non-target organisms, but gene flow occurs to wild relatives growing near potato crops. If stable introgression were to result, the fitness of these wild species could be altered. We therefore transformed the male sterile cultivar Revolucion to provide a genetically modified nematode-resistant potato to evaluate the benefits that this provides until the possibility of stable introgression to wild relatives is determined. Thus, scientific progress is possible without compromise to the precautionary principle. Publication Types: Evaluation Studies Research Support, Non-U.S. Gov't PMID: 15538370 [PubMed - indexed for MEDLINE] 1148: J Agric Food Chem. 2004 Nov 17;52(23):6969-76. Bollgard II cotton: compositional analysis and feeding studies of cottonseed from insect-protected cotton (Gossypium hirsutum L.) producing the Cry1Ac and Cry2Ab2 proteins. Hamilton KA, Pyla PD, Breeze M, Olson T, Li M, Robinson E, Gallagher SP, Sorbet R, Chen Y. Monsanto Company, 800 North Lindbergh Boulevard, St. Louis, Missouri 63167, USA. Bollgard II cotton event 15985 producing the Cry1Ac and Cry2Ab2 proteins has been developed by genetic modification to broaden the spectrum of insects to which the plant is tolerant and to provide an insect resistance management tool to impede the onset of resistance. The purpose of this study was to evaluate the composition and nutrition of Bollgard II cotton, relative to the use for food and animal feed, compared to that of conventional cotton varieties. Compositional analyses were conducted to measure proximate, fiber, amino acid, fatty acid, gossypol, and mineral contents of cottonseed from a total of 14 U.S. field sites over two years. Compositional analysis results showed that the cottonseed and cottonseed oil from Bollgard II cotton were comparable in their composition to those of the conventional control cotton line and other commercial varieties. The composition data are supported by nutritional safety studies conducted with dairy cows, catfish, and quail. Results from these studies showed that Bollgard II performed similarly to the conventional control cotton varieties. These data demonstrate that Bollgard II cotton is compositionally and nutritionally equivalent to conventional cotton varieties. These data support the conclusion that Bollgard II cotton is as safe and nutritious as conventional cotton for food and feed use. PMID: 15537305 [PubMed - indexed for MEDLINE] 1149: J Agric Food Chem. 2004 Nov 17;52(23):6962-8. Qualitative and quantitative evaluation of the genomic DNA extracted from GMO and non-GMO foodstuffs with four different extraction methods. Peano C, Samson MC, Palmieri L, Gulli M, Marmiroli N. Department of Environmental Sciences, University of Parma, Italy. The presence of DNA in foodstuffs derived from or containing genetically modified organisms (GMO) is the basic requirement for labeling of GMO foods in Council Directive 2001/18/CE (Off. J. Eur. Communities 2001, L1 06/2). In this work, four different methods for DNA extraction were evaluated and compared. To rank the different methods, the quality and quantity of DNA extracted from standards, containing known percentages of GMO material and from different food products, were considered. The food products analyzed derived from both soybean and maize and were chosen on the basis of the mechanical, technological, and chemical treatment they had been subjected to during processing. Degree of DNA degradation at various stages of food production was evaluated through the amplification of different DNA fragments belonging to the endogenous genes of both maize and soybean. Genomic DNA was extracted from Roundup Ready soybean and maize MON810 standard flours, according to four different methods, and quantified by real-time Polymerase Chain Reaction (PCR), with the aim of determining the influence of the extraction methods on the DNA quantification through real-time PCR. Publication Types: Research Support, Non-U.S. Gov't PMID: 15537304 [PubMed - indexed for MEDLINE] 1150: EMBO Rep. 2004 Nov;5(11):1031-4. GM plants for your health. The acceptance of GM crops in Europe might grow as soon as the first products to offer direct benefits for consumer health become available. Breithaupt H. PMID: 15520803 [PubMed - indexed for MEDLINE] 1151: Int J Toxicol. 2004;23(5):279-80. Genetically modified foods: why the public frenzy? Role of mainstream news media. Mehendale HM. Publication Types: Editorial PMID: 15513828 [PubMed - indexed for MEDLINE] 1152: J Med Philos. 2004 Jun;29(3):333-50. The precautionary principle and the regulation of U.S. food and drug safety. Soule E. The McDonough School of Business at Georgetown University, Washington, DC 20057, USA. ed.soule@msb.edu This article probes the advisability of regulating U.S. food and drug safety according to the precautionary principle. To do so, a precautionary regulatory regime is formulated on the basis of the beliefs that motivate most proponents of this initiative. That hypothetical regime is critically analyzed on the basis of an actual instantiation of a similarly stylized initiative. It will be argued that the precautionary principle entails regulatory constraints that are apt to violate basis tenets of political legitimacy. The modifications that would change this finding would also change precautionary regulation to the point that it would be indistinguishable from orthodox safety protocols. It is concluded on the basis of its impoverished content that the precautionary principle should not be taken seriously as a formal approach to the regulation of U.S. food and drug safety. Publication Types: Review PMID: 15512976 [PubMed - indexed for MEDLINE] 1153: Mol Nutr Food Res. 2004 Nov;48(6):434-40. Evaluation of the potential allergenicity of the enzyme microbial transglutaminase using the 2001 FAO/WHO Decision Tree. Pedersen MH, Hansen TK, Sten E, Seguro K, Ohtsuka T, Morita A, Bindslev-Jensen C, Poulsen LK. Laboratory of Medical Allergology, Allergy Clinic, National University Hospital, Copenhagen, Denmark. All novel proteins must be assessed for their potential allergenicity before they are introduced into the food market. One method to achieve this is the 2001 FAO/WHO Decision Tree recommended for evaluation of proteins from genetically modified organisms (GMOs). It was the aim of this study to investigate the allergenicity of microbial transglutaminase (m-TG) from Streptoverticillium mobaraense. Amino acid sequence similarity to known allergens, pepsin resistance, and detection of protein binding to specific serum immunoglobulin E (IgE) (RAST) have been evaluated as recommended by the decision tree. Allergenicity in the source material was thought unlikely, since no IgE-mediated allergy to any bacteria has been reported. m-TG is fully degraded after 5 min of pepsin treatment. A database search showed that the enzyme has no homology with known allergens, down to a match of six contiguous amino acids, which meets the requirements of the decision tree. However, there is a match at the five contiguous amino acid level to the major codfish allergen Gad c1. The potential cross reactivity between m-TG and Gad c1 was investigated in RAST using sera from 25 documented cod-allergic patients and an extract of raw codfish. No binding between patient IgE and m-TG was observed. It can be concluded that no safety concerns with regard to the allergenic potential of m-TG were identified. Publication Types: Comparative Study Evaluation Studies PMID: 15508178 [PubMed - indexed for MEDLINE] 1154: Mol Nutr Food Res. 2004 Nov;48(6):413-23. Allergy assessment of foods or ingredients derived from biotechnology, gene-modified organisms, or novel foods. Poulsen LK. Laboratory of Medical Allergology, Allergy Clinic, National University Hospital, Copenhagen, Denmark. lkpallgy@inet.uni2.dk The introduction of novel proteins into foods carries a risk of eliciting allergic reactions in individuals sensitive to the introduced protein and a risk of sensitizing susceptible individuals. No single predictive test exists to perform a hazard assessment in relation to allergenic properties of newly expressed proteins in gene-modified organisms (GMOs). Instead, performance of a weighted risk analysis based on the decision tree approach has been suggested. The individual steps of this analysis comprise sequence homology to known allergens, specific or targeted serum screens for immunoglobulin E (IgE) cross-reactions to known allergens, digestability studies of the proteins in simulated gastric and/or intestinal fluids, and animal studies. These steps are discussed and five examples of risk evaluation of GMOs or novel foods are presented. These include ice-structuring protein derived from fish, microbial transglutaminase, GMO-soybeans, amylase and the Nangai nut. Publication Types: Review PMID: 15508176 [PubMed - indexed for MEDLINE] 1155: Biotechnol Annu Rev. 2004;10:85-122. Public health issues related with the consumption of food obtained from genetically modified organisms. Paparini A, Romano-Spica V. University of Rome Foro Italico (IUSM), Rome, Italy. Genetically Modified Organisms (GMOs) are a fact of modern agriculture and a major field of discussion in biotechnology. As science incessantly achieves innovative and unexpected breakthroughs, new medical, political, ethical and religious debates arise over the production and consumption of transgenic organisms. Despite no described medical condition being directly associated with a diet including approved GM crops in large exposed populations such as 300,000,000 Americans and a billion Chinese, public opinion seems to look at this new technology with either growing concern or even disapproval. It is generally recognized that a high level of vigilance is necessary and highly desirable, but it should also be considered that GMOs are a promising new challenge for the III Millennium societies, with remarkable impact on many disciplines and fields related to biotechnology. To acquire a basic knowledge on GMO production, GM-food consumption, GMO interaction with humans and environment is of primary importance for risk assessment. It requires availability of clear data and results from rigorous experiments. This review will focus on public health risks related with a GMO-containing diet. The objective is to summarize state of the art research, provide fundamental technical information, point out problems and perspectives, and make available essential tools for further research. Are GMO based industries and GMO-derived foods safe to human health? Can we consider both social, ethical and public health issues by means of a constant and effective monitoring of the food chain and by a clear, informative labeling of the products? Which are the so far characterized or alleged hazards of GMOs? And, most importantly, are these hazards actual, potential or merely contrived? Several questions remain open; answers and solutions belong to science, to politics and to the personal opinion of each social subject. Publication Types: Review PMID: 15504704 [PubMed - indexed for MEDLINE] 1156: Proc Biol Sci. 2004 Aug 7;271 Suppl 5:S350-2. Growth hormone transgenic salmon pay for growth potential with increased predation mortality. Sundström LF, Lõhmus M, Johnsson JI, Devlin RH. Department of Zoology, Göteborg University, SE-405 30 Göteborg, Sweden. Recent advances in gene technology have been applied to create fast-growing transgenic fish, which are of great commercial interest owing to their potential to shorten production cycles and increase food production. However, there is growing concern and speculation over the impact that escaped growth hormone (GH)-transgenic fish may have on the natural environment. To predict these risks it is crucial to obtain empirical data on the relative fitness of transgenic and non-transgenic fish under nature-like conditions. Using landscaped stream aquaria with live food and predators, we show that the predation mortality of newly hatched GH-transgenic coho salmon fry (Oncorhynchus kisutch) is much higher than in non-transgenic conspecifics, and that this difference is amplified when food abundance decreases. The growth rate of transgenic and non-transgenic fish is similar at high food levels, whereas transgenic fish grow more slowly than non-transgenic fish when food abundance is reduced. Our results suggest that the fitness of young GH-transgenic coho salmon in the wild will be determined by both predation pressure and food availability. Publication Types: Comparative Study Research Support, Non-U.S. Gov't PMID: 15504015 [PubMed - indexed for MEDLINE] 1157: Mol Med. 2004 Jan-Jun;10(1-6):36-44. Attenuated hippocampus-dependent learning and memory decline in transgenic TgAPPswe Fischer-344 rats. Ruiz-Opazo N, Kosik KS, Lopez LV, Bagamasbad P, Ponce LR, Herrera VL. Section of Molecular Medicine, Department of Medicine, Boston University School of Medicine, Boston, Massachusetts 02118, USA. Alzheimer's disease (AD) is characterized by increased beta amyloid (Abeta) levels, extracellular Abeta deposits in senile plaques, neurofibrillary tangles, and neuronal loss. However, the physiological role of normal levels of Abeta and its parent protein, the amyloid precursor protein (APP) are unknown. Here we report that low-level transgenic (Tg) expression of the Swedish APP mutant gene (APPswe) in Fischer-344 rats results in attenuated age-dependent cognitive performance decline in 2 hippocampus-dependent learning and memory tasks compared with age-matched nontransgenic Fischer-344 controls. TgAPPswe rats exhibit mild increases in brain APP mRNA (56.8%), Abeta-42 (21%), and Abeta-40 (6.1%) peptide levels at 12 mo of age, with no extracellular Abeta deposits or senile plaques at 6, 12, and 18 mo of age, whereas 3- to 6-fold increases in Abeta levels are detected in plaque-positive human AD patients and transgenic mouse models. The data support the hypothesis that a threshold paradigm underlies Abeta-related pathology, below which APP expression may play a physiological role in specific hippocampus-dependent tasks, most likely related to its neurotrophic role. Publication Types: Research Support, N.I.H., Extramural Research Support, U.S. Gov't, P.H.S. PMID: 15502881 [PubMed - indexed for MEDLINE] 1158: Nurs Outlook. 2004 Sep-Oct;52(5):262-6. "This food may contain ..." What nurses should know about genetically engineered foods. Whitney SL, Maltby HJ, Carr JM. College of Nursing and Health Sciences, University of Vermont, 220 Rowell, Burlington, VT 05405, USA. Stuart.Whitney@uvm.edu Genetic engineering has been in existence since 1973. The process involves placing genetic DNA from one organism into another. Genetically engineered organisms (GEOs) are the name given to such new species of plants created through this process. Proponents of GEOs assert that foods we are now able to produce have greater nutritional value, longer shelf life, better appearance, taste and smell. There are positive benefits to genetic engineering of plants and animals. A growing concern for the health safety of genetically engineered plants and foods is developing among the cautious. The purpose of this article is to define genetic engineering, present benefits and risks, describe the impact on human health, and address implications for nursing. Publication Types: Review PMID: 15499316 [PubMed - indexed for MEDLINE] 1159: FEBS Lett. 2004 Oct 22;576(3):477-80. Expression proteomics identifies biochemical adaptations and defense responses in transgenic plants with perturbed polyamine metabolism. Franceschetti M, Perry B, Thompson B, Hanfrey C, Michael AJ. Division of Food Safety Science, Institute of Food Research, Norwich Research Park, Colney, Norwich, NR4 7UA, UK. Soluble proteins from leaves of transgenic tobacco plants with perturbed polyamine metabolism, caused by S-adenosylmethionine decarboxylase overexpression, were analysed by comparative proteomics. A group of proteins was found to be increasingly repressed, in parallel with the degree of polyamine perturbation, in each of the three independent transgenic lines. These were identified as isoforms of chloroplast ribonucleoproteins, known to be involved in chloroplast mRNA stability, processing and translation. Another group of eight proteins strongly induced in the most metabolically perturbed line was identified as multiple, uncharacterised isoforms of the defense protein PR-1, a known marker for systemic acquired resistance. Publication Types: Research Support, Non-U.S. Gov't PMID: 15498583 [PubMed - indexed for MEDLINE] 1160: QJM. 2004 Nov;97(11):705-16. Biopharmaceuticals derived from genetically modified plants. Goldstein DA, Thomas JA. Monsanto Company A2NE, 800 N. Lindbergh Blvd, St Louis, MO 63167, USA. daniel.a.goldstein@monsanto.com Modern biotechnology has resulted in a resurgence of interest in the production of new therapeutic agents using botanical sources. With nearly 500 biotechnology products approved or in development globally, and with production capacity limited, the need for efficient means of therapeutic protein production is apparent. Through genetic engineering, plants can now be used to produce pharmacologically active proteins, including mammalian antibodies, blood product substitutes, vaccines, hormones, cytokines, and a variety of other therapeutic agents. Efficient biopharmaceutical production in plants involves the proper selection of host plant and gene expression system, including a decision as to whether a food crop or a non-food crop is more appropriate. Product safety issues relevant to patients, pharmaceutical workers, and the general public must be addressed, and proper regulation and regulatory oversight must be in place prior to commercial plant-based biopharmaceutical production. Plant production of pharmaceuticals holds great potential, and may become an important production system for a variety of new biopharmaceutical products. Publication Types: Review PMID: 15496527 [PubMed - indexed for MEDLINE] 1161: Curr Atheroscler Rep. 2004 Nov;6(6):468-76. Dietary fats and oils: technologies for improving cardiovascular health. Flickinger BD, Huth PJ. ADM Reserach, James R. Randall Research Center, Decatur, IL 62521, USA. flickinger@admworld.com The role of dietary lipids in the etiology of coronary heart disease (CHD) continues to evolve as we gain a better understanding of the metabolic effects of individual fatty acids and their impact on surrogate markers of risk. A recent meta-analysis of 60 human studies suggests that for each 1% energy replacement of carbohydrates in the diet with saturated fat or trans fat, serum low-density lipoprotein cholesterol concentrations increase by 0.032 (1.23 mg/dL) and 0.04 mmol/L (1.54 mg/dL), respectively. Current dietary recommendations to keep saturated fat and trans fat intake as low as possible, and to increase the intake of cis mono-unsaturated and polyunsaturated fatty acids, as well as growing recognition of these recommendations by consumers and food regulatory agencies in the United States, have been major driving forces for the edible oil industry and food manufacturers to develop alternative fats and oils with nutritionally improved fatty acid compositions. As solutions for use of trans fatty acids are being sought, oilseeds with modified fatty acid compositions are being viewed as a means to provide such solutions. Additionally, oilseeds with modified fatty acid composition, such as enhanced content of long-chain omega-3 fatty acids or conjugated linoleic acid, have been developed as a way to increase delivery of these fatty acids directly into the food supply or indirectly as use for feed ingredients for livestock. New processing technologies are being utilized around the world to create dietary fats and oils with specific physiologic functions relevant to risk factors for cardiovascular disease. Publication Types: Review PMID: 15485593 [PubMed - indexed for MEDLINE] 1162: Int J Occup Environ Health. 2004 Jul-Sep;10(3):296-303. Genetically modified cotton and farmers' health in China. Hossain F, Pray CE, Lu Y, Huang J, Fan C, Hu R. Department of Agricultural, Food and Resource Economics, Rutgers University, New Brunswick, New Jersey 08901-8520, USA. This study provides the first evidence of a direct link between the adoption of a genetically modified (GM) crop and improvements in human health. Estimation of the impact of Bacillus thuringiensis (Bt) cotton adoption on pesticide use from data from a survey of cotton farmers in northern China, 1999-2001, showed that Bt cotton adoption reduced pesticide use. Assessment of a health-production function showed that predicted pesticide use had a positive impact on poisoning incidence. Taken together, these results indicate that the adoption of Bt cotton can substantially reduce the risk and the incidence of poisonings. Publication Types: Research Support, Non-U.S. Gov't PMID: 15473084 [PubMed - indexed for MEDLINE] 1163: Nat Biotechnol. 2004 Oct;22(10):1207-8. Comment on: Nat Biotechnol. 2004 Jul;22(7):811-2. Putting Cartagena into practice. Watanabe KN, Taeb M, Okusu H. Publication Types: Comment Letter PMID: 15470448 [PubMed - indexed for MEDLINE] 1164: Tradition. 2003 Summer;37(2):66-87. Survey of recent halakhic periodical literature: genetic engineering. Bleich JD. PMID: 15468505 [PubMed - indexed for MEDLINE] 1165: Not Polit. 2001;17(62):51-76. Expert discourses of risk and ethics on genetically manipulated organisms: the weaving of public alienation. Wynne B. CSEC, Lancaster University, UK. PMID: 15468486 [PubMed - indexed for MEDLINE] 1166: Not Polit. 2001;17(62):22-33. Knowledge and the governance of biotechnology. Doubleday R. Harvard University, USA. PMID: 15468484 [PubMed - indexed for MEDLINE] 1167: Expert Opin Biol Ther. 2004 Oct;4(10):1565-8. Transgenic plant-derived pharmaceuticals - the practical approach? Yano A, Takekoshi M. Production of biopharmaceuticals in transgenic plants would involve the creation of a new industry. Those transgenic plants, including staple food crops, could provide many benefits to people all over the world. However, the new industry might require a strict regulation system. It is probable that such a strict system would not be acceptable to Japan or to most developing countries. Many countries should use non-food crops for production of biopharmaceuticals and take on more simple systems. The new industry must develop strategies for promoting the benefits of transgenic plant-derived biopharmaceuticals on both the domestic and worldwide scales. Publication Types: Editorial Research Support, Non-U.S. Gov't PMID: 15461567 [PubMed - indexed for MEDLINE] 1168: Tijdschr Diergeneeskd. 2004 Sep 1;129(17):553. [Do consumers trust their food?] [Article in Dutch] [No authors listed] PMID: 15461373 [PubMed - indexed for MEDLINE] 1169: Wei Sheng Yan Jiu. 2004 Jul;33(4):502-4. [Methods of hygromycin B phosphotransferase activity assay in transgenic plant] [Article in Chinese] Zhuo Q, Yang X. Institute for Nutrition and Food Safety, Chinese Center for Disease Control and Prevention, Beijing 100050, China. Hygromycin B phosphotransferase (HPT) is a widely used selectable marker protein of transgenic plant. Detection of its activity is critical to studies on the development of various transgenic plants, silence of inserted gene, marker-free system development and safety assessment of transgenic food. In this paper, several methods for detecting the activity of this enzyme were reviewed. Publication Types: English Abstract Research Support, Non-U.S. Gov't Review PMID: 15461291 [PubMed - indexed for MEDLINE] 1170: Wei Sheng Yan Jiu. 2004 Jul;33(4):437-9. [Preparation and application of monoclonal antibodies against cowpea trypsin inhibitor] [Article in Chinese] Chen X, Yang L, Yang X, Piao J. National Institute For Nutrition and Food Safety, Chinese Center for Disease Prevention and Control, Beijing. OBJECTIVE: To detect cowpea trypsin inhibitor (CpTI) in transgenic rice leaves and build a possible method to assay transgenic plant with CpTI. METHODS: Nowadays gene assay is the most useful method to detect transgenic plants. But false positive may be got in gene assay and effective protein may not be produced. So assay of foreign protein is the most direct and conformable method. Monoclonal antibodies were got through traditional method of preparation. RESULTS: Three monoclonal antibodies were got. Detection of CpTI in transgenic rice leaves with these three antibodies mixes through Western blotting was conducted and the result was satisfying. CONCLUSION: The method of using the mixes of three antibodies to detect the CpTI in plants through Western blotting was satisfying. Publication Types: English Abstract Research Support, Non-U.S. Gov't PMID: 15461269 [PubMed - indexed for MEDLINE] 1171: J Agric Food Chem. 2004 Oct 6;52(20):6075-85. NMR and HPLC-UV profiling of potatoes with genetic modifications to metabolic pathways. Defernez M, Gunning YM, Parr AJ, Shepherd LV, Davies HV, Colquhoun IJ. Institute of Food Research, Norwich Research Park, Colney, Norwich NR4 7UA, United Kingdom. Metabolite profiling has been carried out to assess the compositional changes occurring in potato tubers after genetic modifications have been made to different metabolic pathways. Most major features in the (1)H NMR and HPLC-UV profiles of tuber extracts have been assigned. About 40 GM lines and controls belonging to 4 groups of samples (derived from cv. Record or cv. Desirée and modified in primary carbon metabolism, starch synthesis, glycoprotein processing, or polyamine/ethylene metabolism) were analyzed. Differences were assessed at the level of whole profiles (by PCA) or individual compounds (by ANOVA). The most obvious differences seen in both NMR and HPLC-UV profiles were between the two varieties. There were also significant differences between two of the four Desirée GM lines with modified polyamine metabolism and their controls. Compounds notably affected were proline, trigonelline, and numerous phenolics. However, that modification gave rise to a very abnormal phenotype. Certain lines from the other groups had several compounds present in significantly higher or lower amounts compared to the control, but the differences in mean values amounted to no more than a 2-3-fold change: in the context of variability in the whole data set, such changes did not appear to be important. Publication Types: Research Support, Non-U.S. Gov't PMID: 15453669 [PubMed - indexed for MEDLINE] 1172: J Biol Chem. 2004 Dec 3;279(49):50781-9. Epub 2004 Sep 21. Drosophila short neuropeptide F regulates food intake and body size. Lee KS, You KH, Choo JK, Han YM, Yu K. Laboratory of Development and Differentiation, Korea Research Institute of Bioscience and Biotechnology, 52 Eoun-dong, Yusong-gu, Daejeon, 395-333, Korea. Neuropeptides regulate a wide range of animal behavior including food consumption, circadian rhythms, and anxiety. Recently, Drosophila neuropeptide F, which is the homolog of the vertebrate neuropeptide Y, was cloned, and the function of Drosophila neuropeptide F in feeding behaviors was well characterized. However, the function of the structurally related short neuropeptide F (sNPF) was unknown. Here, we report the cloning, RNA, and peptide localizations, and functional characterizations of the Drosophila sNPF gene. The sNPF gene encodes the preprotein containing putative RLRF amide peptides and was expressed in the nervous system of late stage embryos and larvae. The embryonic and larval localization of the sNPF peptide in the nervous systems revealed the larval central nervous system neural circuit from the neurons in the brain to thoracic axons and to connective axons in the ventral ganglion. In the adult brain, the sNPF peptide was localized in the medulla and the mushroom body. However, the sNPF peptide was not detected in the gut. The sNPF mRNA and the peptide were expressed during all developmental stages from embryo to adult. From the feeding assay, the gain-of-function sNPF mutants expressed in nervous systems promoted food intake, whereas the loss-of-function mutants suppressed food intake. Also, sNPF overexpression in nervous systems produced bigger and heavier flies. These findings indicate that the sNPF is expressed in the nervous systems to control food intake and regulate body size in Drosophila melanogaster. Publication Types: Research Support, Non-U.S. Gov't PMID: 15385546 [PubMed - indexed for MEDLINE] 1173: Am J Law Med. 2004;30(2-3):371-404. Consuming (f)ears of corn: public health and biopharming. Bratspies RM. CUNY School of Law, USA. PMID: 15382759 [PubMed - indexed for MEDLINE] 1174: Bundesgesundheitsblatt Gesundheitsforschung Gesundheitsschutz. 2004 Sep;47(9):826-33. [Genetically modified plants and food safety. State of the art and discussion in the European Union] [Article in German] Schauzu M. Bundesinstitut für Risikobewertung, Berlin. m.schauzu@bfr.bund.de Placing genetically modified (GM) plants and derived products on the European Union's (EU) market has been regulated by a Community Directive since 1990. This directive was complemented by a regulation specific for genetically modified and other novel foods in 1997. Specific labelling requirements have been applicable for GM foods since 1998. The law requires a pre-market safety assessment for which criteria have been elaborated and continuously adapted in accordance with the state of the art by national and international bodies and organisations. Consequently, only genetically modified products that have been demonstrated to be as safe as their conventional counterparts can be commercialized. However, the poor acceptance of genetically modified foods has led to a de facto moratorium since 1998. It is based on the lack of a qualified majority of EU member states necessary for authorization to place genetically modified plants and derived foods on the market. New Community Regulations are intended to end this moratorium by providing a harmonized and transparent safety assessment, a centralised authorization procedure, extended labelling provisions and a traceability system for genetically modified organisms (GMO) and derived food and feed. Publication Types: English Abstract Review PMID: 15378169 [PubMed - indexed for MEDLINE] 1175: Zhonghua Yu Fang Yi Xue Za Zhi. 2003 Mar;37(2):133-5. [Assessment of the allergenic potential of genetically modified food] [Article in Chinese] Xu M. School of Food and Biologic engineering, Hangzhou Collage of Commercial, Hangzhou 310035, China Publication Types: Review PMID: 15376370 [PubMed - indexed for MEDLINE] 1176: BMC Bioinformatics. 2004 Sep 16;5:133. Allermatch, a webtool for the prediction of potential allergenicity according to current FAO/WHO Codex alimentarius guidelines. Fiers MW, Kleter GA, Nijland H, Peijnenburg AA, Nap JP, van Ham RC. BACKGROUND: Novel proteins entering the food chain, for example by genetic modification of plants, have to be tested for allergenicity. Allermatch http://allermatch.org is a webtool for the efficient and standardized prediction of potential allergenicity of proteins and peptides according to the current recommendations of the FAO/WHO Expert Consultation, as outlined in the Codex alimentarius. DESCRIPTION: A query amino acid sequence is compared with all known allergenic proteins retrieved from the protein databases using a sliding window approach. This identifies stretches of 80 amino acids with more than 35% similarity or small identical stretches of at least six amino acids. The outcome of the analysis is presented in a concise format. The predictive performance of the FAO/WHO criteria is evaluated by screening sets of allergens and non-allergens against the Allermatch databases. Besides correct predictions, both methods are shown to generate false positive and false negative hits and the outcomes should therefore be combined with other methods of allergenicity assessment, as advised by the FAO/WHO. CONCLUSIONS: Allermatch provides an accessible, efficient, and useful webtool for analysis of potential allergenicity of proteins introduced in genetically modified food prior to market release that complies with current FAO/WHO guidelines. PMID: 15373946 [PubMed - indexed for MEDLINE] 1177: Nature. 2004 Sep 16;431(7006):238-43. Comment in: Nature. 2004 Nov 25;432(7016):439. Head to head: Bush vs Kerry. Macilwain C, Bush GW, Kerry J. Publication Types: Interview News PMID: 15372001 [PubMed - indexed for MEDLINE] 1178: Transgenic Res. 2004 Jun;13(3):295-8. Fimbrial subunit protein FaeG expressed in transgenic tobacco inhibits the binding of F4ac enterotoxigenic Escherichia coli to porcine enterocytes. Joensuu JJ, Kotiaho M, Riipi T, Snoeck V, Palva ET, Teeri TH, Lång H, Cox E, Goddeeris BM, Niklander-Teeri V. Department of Applied Biology, P.O. Box 27, FIN-00014 University of Helsinki, Finland. jussi.joensuu@helsinki.fi Plants offer a promising alternative for the production of foreign proteins for pharmaceutical purposes in tissues that are consumed as food and/or feed. Our long-term strategy is to develop edible vaccines against piglet diarrhoea caused by enterotoxigenic Escherichia coli (F4 ETEC) in feed plants. In this work, we isolated a gene, faeG, encoding for a major F4ac fimbrial subunit protein. Our goal was to test whether the FaeG protein, when isolated from its fimbrial background and produced in a plant cell, would retain the key properties of an oral vaccine, that is, stability in gastrointestinal conditions, binding to intestinal receptors and inhibition of the F4 ETEC attachment. For this purpose, tobacco was first transformed with a faeG construct that included a transit peptide encoding sequence to target the FaeG protein to the chloroplast. The best transgenic lines produced FaeG protein in amounts of 1% total soluble protein. The stability of the plant-produced FaeG was tested in fluids simulating piglet gastric (SGF) and intestinal (SIF) conditions. Plant-produced FaeG proved to be stable up to 2 h under these conditions. The binding and inhibition properties were tested with isolated piglet villi. These results showed that the plant-produced FaeG could bind to the receptors on the villi and subsequently inhibit F4 ETEC binding in a dose-dependent manner. Thus, the first two prerequisites for the development of an oral vaccine have been met. Publication Types: Comparative Study Research Support, Non-U.S. Gov't PMID: 15359606 [PubMed - indexed for MEDLINE] 1179: Planta. 2005 Jan;220(3):455-64. Epub 2004 Sep 4. Expression of poly-3-(R)-hydroxyalkanoate (PHA) polymerase and acyl-CoA-transacylase in plastids of transgenic potato leads to the synthesis of a hydrophobic polymer, presumably medium-chain-length PHAs. Romano A, van der Plas LH, Witholt B, Eggink G, Mooibroek H. Agrotechnology and Food Innovations B.V., Department of Bioconversion, Wageningen University and Research Centre, POB 17, 6700 AA, Wageningen, The Netherlands. Medium-chain-length poly-3-(R)-hydroxyalkanoates (mcl-PHAs) belong to the group of microbial polyesters. The minimum gene-set for the accumulation of mcl-PHAs from de novo fatty acid biosynthesis has been identified in prokaryotes as consisting of the Pha-C1 polymerase and the ACP-CoA-transacylase. In this paper, the synthesis of mcl-PHAs has been attempted in transgenic potato (Solanum tuberosum L.) using the same set of genes that were introduced into potato by particle bombardment. Polymer contents of transgenic lines were analysed by gas chromatography and by a new simple method employing a size-exclusion filter column. The expression of the Pha-C1 polymerase and the ACP-CoA-transacylase in the plastids of transgenic potato led to the synthesis of a hydrophobic polymer composed of mcl-hydroxy-fatty acids with carbon chain lengths ranging from C-6 to C-12 in leaves of the selected transgenic lines. We strongly suggest that the polymer observed consists of mcl-PHAs and that this report establishes for the first time a possible route for the production of mcl-PHAs from de novo fatty acid biosynthesis in plants. Publication Types: Research Support, Non-U.S. Gov't PMID: 15351883 [PubMed - indexed for MEDLINE] 1180: Nat Biotechnol. 2004 Sep;22(9):1062. NAS issues mixed message on unintended effects of GM food. Fox JL. Publication Types: News PMID: 15340456 [PubMed - indexed for MEDLINE] 1181: Nat Biotechnol. 2004 Sep;22(9):1055. Comment in: Nat Biotechnol. 2004 Dec;22(12):1501; discussion 1501. Orphans at the window. [No authors listed] Publication Types: Editorial PMID: 15340451 [PubMed - indexed for MEDLINE] 1182: Nat Med. 2004 Sep;10(9):881. Edible vaccines not ready for main course. Vermij P. Publication Types: News PMID: 15340395 [PubMed - indexed for MEDLINE] 1183: Poult Sci. 2004 Aug;83(8):1325-34. Evaluation of broiler performance when fed Roundup-Ready wheat (event MON 71800), control, and commercial wheat varieties. Kan CA, Hartnell GF. Animal Sciences Group, Nutrition and Food, 8200 AB, Lelystad, The Netherlands. kees.kan@wur.nl We evaluated the nutritional value of broiler diets containing approximately 40% wheat grain from Roundup Ready wheat (MON 71800), its similar nontransgenic control (MON 71900), or reference commercial wheat varieties. The feeding trial lasted 40 d, and each treatment consisted of 10 replicates of 1-d-old Ross 308 broilers (5 pens of males and 5 pens of females). Each pen contained 12 birds, and at d 13 birds were randomly removed until 9 birds remained. Body weight and feed intake were measured on pen basis at 40 d. At d 41, four broilers per pen were slaughtered. The carcasses were dissected, and cut-up yields were determined. Dry matter, protein, and fat contents of breast meat were determined. The data were analyzed by an ANOVA procedure. The BW and feed conversion at d 40 averaged 2,450 g and 1.52, respectively. There were no significant treatment x sex interactions, except for evisceration yield with significant differences (P < 0.05) in yield between birds fed 2 commercial wheat varieties. Data for final BW, feed conversion, carcass yield, and breast meat were not statistically different (P < 0.05) between broilers fed MON 71800 or MON 71900 or the population of birds fed commercial wheat varieties, except a lower carcass yield at d 41 for birds fed the nontransgenic control wheat. Thus MON 71800 was nutritionally equivalent to nongenetically modified wheat varieties when fed to broilers. Publication Types: Clinical Trial Randomized Controlled Trial PMID: 15339007 [PubMed - indexed for MEDLINE] 1184: Carbohydr Res. 2004 Sep 13;339(13):2233-9. Synergistic interactions between the genetically modified bacterial polysaccharide P2 and carob or konjac mannan. Ridout M, Cairns P, Brownsey G, Morris V. Institute of Food Research, Food Material Science, Norwich Research Park, Colney, Norwich NR4 7UA, UK. Rheological studies have confirmed that the bacterial polysaccharide P2, a genetically modified variant of the Acetobacter xylinum polysaccharide acetan, undergoes synergistic gelation with either of the plant polysaccharides carob or konjac mannan. X-ray fibre diffraction data shows that P2 can form a 5-fold helical structure of pitch 4.7nm and an axial rise per disaccharide repeat of 0.92nm. Optical rotation data demonstrate that P2 undergoes a coil-helix transition in solution and that deacylation enhances the stability of the helical structure in solution. Studies made on mixtures prepared at different temperatures and ionic strengths suggest that denaturation of the P2 helix favours interaction and gelation. Deacetylation of P2 enhances gelation. X-ray diffraction data for oriented fibres prepared from deacetylated P2-konjac mannan mixed films reveal a 6-fold helical structure of pitch 5.54nm with an axial rise per disaccharide repeat also of 0.92nm. This mixed helix provides direct evidence for binding between the two polysaccharides. P2 contains two sites of acetylation: one on the backbone and one on the sidechain. The former site of acetylation inhibits helix formation for P2. It is suggested that this site of acetylation also inhibits formation of the mixed helix, explaining the enhanced gelation of mixtures on deacetylation. Publication Types: Research Support, Non-U.S. Gov't PMID: 15337451 [PubMed - indexed for MEDLINE] 1185: Neuropeptides. 2004 Aug;38(4):189-200. NPY and Y receptors: lessons from transgenic and knockout models. Lin S, Boey D, Herzog H. Neurobiology Program, Garvan Institute of Medical Research, 384 Victoria Street, Darlinghurst Sydney, NSW 2010, Australia. Neuropeptide Y (NPY) in the central nervous system is a major regulator of food consumption and energy homeostasis. It also regulates blood pressure, induces anxiolysis, enhances memory retention, affects circadian rhythms and modulates hormone release. Five Y receptors (Y1, Y2, Y4, Y5 and Y6) are known to mediate the action of NPY and its two other family members, peptide YY (PYY) and pancreatic polypeptide (PP). Increased NPY signaling due to elevated NPY expression in the hypothalamus leads to the development of obesity and its related phenotypes, Type II diabetes and cardiovascular disease. Dysregulation in NPY signaling also causes alterations in bone formation, alcohol consumption and seizure susceptibility. The large number of Y receptors has made it difficult to delineate their individual contributions to these physiological processes. However, recent studies analysing NPY and Y receptor overexpressing and knockout models have started to unravel some of the different functions of these Y receptors. Particularly, the use of conditional knockout models has made it possible to pinpoint a specific function to an individual Y receptor in a particular location. Publication Types: Review PMID: 15337371 [PubMed - indexed for MEDLINE] 1186: Ceylon Med J. 2004 Jun;49(2):44-6. Genetically modified food: friend or foe? Perera BJ. Lady Ridgeway Hospital for Children, Sri Lanka. bjcp@sltnet.lk Publication Types: Review PMID: 15334797 [PubMed - indexed for MEDLINE] 1187: Indian J Dent Res. 2003 Oct-Dec;14(4):284-8. Genetic engineering and dental caries. Agarwal S, Pandit IK, Srivastava N, Gugnani N. Department of Pedodontics and Preventive Dentistry, DAVO Dental College and Hospital, Yamunanagar, Haryana--135001. Dental caries, a multifactorial disease requires four principle factor: the host, the microflora, the substrate & time for its occurrence and can be prevented or managed by elimination/modification of either of the above factors. The conventional preventive measure being followed for long time for the dental caries are not successful to the desirable extent due to their non avaibailaballity in the rural areas, lack of awareness & inaccessibility of dental services. Therefore, the focus has now been shifted to submicroscopic level to ensure that these measures can be reached to the farthest areas & each & every member of the population is benefitted. Few of the measures taken are. i) Genetically modifying the S. Mutans: ii) Searching The antagonist peptides to work against the specific enzyme system (Glucosyltransferase) of S. Mutans. iii) Changing the oral environment by those Genetically modified organisms that will produce bases (instead of acids) & these bases provides a milieu favoring remineralization. This paper discusses various ways in which genetically modified strains of microogranisms or genetically modified strains of microogranisms of genetically modified foods can help in the prevention of caries. PMID: 15328998 [PubMed - indexed for MEDLINE] 1188: Public Underst Sci. 2004 Apr;13(2):155-75. Dynamics of list-server discussion on genetically modified foods. Triunfol ML, Hines PJ. Associate editor at the American Assocation for the Advancement of Science (AAAS). mtriunfo@aaas.org Computer-mediated discussion lists, or list-servers, are popular tools in settings ranging from professional to personal to educational. A discussion list on genetically modified food (GMF) was created in September 2000 as part of the Forum on Genetically Modified Food developed by Science Controversies: Online Partnerships in Education (SCOPE), an educational project that uses computer resources to aid research and learning around unresolved scientific questions. The discussion list "GMF-Science" was actively supported from January 2001 to May 2002. The GMF-Science list welcomed anyone interested in discussing the controversies surrounding GMF. Here, we analyze the dynamics of the discussions and how the GMF-Science list may contribute to learning. Activity on the GMF-Science discussion list reflected some but not all the controversies that were appearing in more traditional publication formats, broached other topics not well represented in the published literature, and tended to leave undiscussed the more technical research developments. Publication Types: Historical Article PMID: 15323060 [PubMed - indexed for MEDLINE] 1189: Curr Med Chem Cardiovasc Hematol Agents. 2003 Jun;1(2):197-202. Design of a genetically modified soybean protein preventing hypertension based on an anti-hypertensive peptide derived from ovalbumin. Matoba N, Yamada Y, Yoshikawa M. Division of Food Bioscience and Biotechnology, Graduate School of Agriculture, Kyoto University, Uji, Kyoto 611-0011, Japan. Food proteins can be a source of various bioactive peptides including such possessing anti-hypertensive activity. While most orally active anti-hypertensive peptides derived from food proteins inhibit the angiotensin I-converting enzyme (ACE), ovokinin (2-7) (RADHPF), a peptide isolated from a chymotryptic digest of ovalbumin, has been shown to induce nitric oxide-dependent vasorelaxation in an isolated mesenteric artery as well as anti-hypertensive effect after oral administration in spontaneously hypertensive rats (SHRs). Rational amino acid replacement lead to the ovokinin (2-7) analog, RPLKPW, which had the highest anti-hypertensive activity among the tested peptides. Furthermore, oral administration (0.1 mg/kg) of the peptide lowered the blood pressure of SHR but not of normotensive Wistar-Kyoto (WKY) rats. In order to develop a novel use of this potent anti-hypertensive peptide for prevention of hypertension, RPLKPW has been genetically introduced into the homologous sequences in soybean beta-conglycinin alpha' subunit by site-directed mutagenesis. The recombinant RPLKPW-incorporated alpha' subunit expressed in E. coil has been shown to exert anti-hypertensive activity after oral administration in SHR. Thus, RPLKPW-incorporated alpha' subunit is the first example of a genetically modified food protein possessing physiological activity based on a bioactive peptide. Publication Types: Research Support, Non-U.S. Gov't PMID: 15320699 [PubMed - indexed for MEDLINE] 1190: Public Underst Sci. 2004 Apr;13(2):131-53. Preferences need no inferences, once again: germinal elements in the public perceptions of genetically modified foods in Colombia. Parales-Quenza CJ. Universidad del Rosario, Calle 14 6-25 Bogotá D.C., Colombia. cparales@urosario.edu.co This paper explores the public perceptions of genetically modified foods in Colombia in a phase considered germinal: the topic was too novel at the time of research. The analysis covers media, informal conversations, and the word associations made by a sample of residents in the city of Bogotá. The results show that the public capability of associating with the topic, even intuitively, is due to the availability of culture themes, the primary categories that help conceptual elaborations, and the construction of common-sense theories. Three themes are proposed: natural/artificial, tradition/change, and health/disease. It is argued that cultural themes are not only cognitive, but also affectively laden entities, which explains the evaluative force expressed by social beliefs. Acknowledging the relevance of the non-attitude thesis, the author suggests that people associate novel objects with latent cultural meanings, explaining why words, images, and metaphors are readily available in elaborating social knowledge. Publication Types: Historical Article PMID: 15320334 [PubMed - indexed for MEDLINE] 1191: Bioinformatics. 2005 Jan 1;21(1):39-50. Epub 2004 Aug 19. Supervised identification of allergen-representative peptides for in silico detection of potentially allergenic proteins. Björklund AK, Soeria-Atmadja D, Zorzet A, Hammerling U, Gustafsson MG. Division of Toxicology, National Food Administration, P.O. Box 622, SE-751 26 Uppsala, Sweden. MOTIVATION: Identification of potentially allergenic proteins is needed for the safety assessment of genetically modified foods, certain pharmaceuticals and various other products on the consumer market. Current methods in bioinformatic allergology exploit common features among allergens for the detection of amino acid sequences of potentially allergenic proteins. Features for identification still unexplored include the motifs occurring commonly in allergens, but rarely in ordinary proteins. In this paper, we present an algorithm for the identification of such motifs with the purpose of biocomputational detection of amino acid sequences of potential allergens. RESULTS: Identification of allergen-representative peptides (ARPs) with low or no occurrence in proteins lacking allergenic properties is the essential component of our new method, designated DASARP (Detection based on Automated Selection of Allergen-Representative Peptide). This approach consistently outperforms the criterion based on identical peptide match for predicting allergenicity recommended by ILSI/IFBC and FAO/WHO and shows results comparable to the alignment-based criterion as outlined by FAO/WHO. AVAILABILITY: The detection software and the ARP set needed for the analysis of a query protein reported here are properties of the Swedish National Food Agency and are available upon request. The protein sequence sets used in this work are publicly available on http://www.slv.se/templatesSLV/SLV_Page____9343.asp. Allergenicity assessment for specific protein sequences of interest is also possible via ulfh@slv.se Publication Types: Comparative Study Evaluation Studies Research Support, Non-U.S. Gov't Validation Studies PMID: 15319257 [PubMed - indexed for MEDLINE] 1192: Reprod Fertil Dev. 2004;16(4):465-70. State of the art in the production of transgenic goats. Baldassarre H, Wang B, Keefer CL, Lazaris A, Karatzas CN. Nexia Biotechnologies Inc., Dorion-Vaudreuil, Quebec, Canada. hbaldassarre@nexiabiotech.com This review summarises recent advances in the field of transgenic goats for the purpose of producing recombinant proteins in their milk. Production of transgenic goats via pronuclear microinjection of DNA expression vectors has been the traditional method, but this results in low efficiencies. Somatic cell nuclear transfer has dramatically improved efficiencies in rates of transgenesis. Characterisation of transfected cells in vitro before use in nuclear transfer guarantees that kids born are transgenic and of predetermined gender. Using these platform technologies, several recombinant proteins of commercial interest have been produced, although none of them has yet gained marketing approval. Before these technologies are implemented in goat improvement programmes, efficiencies must be improved, costs reduced, and regulatory approval obtained for the marketing of food products derived from such animals. Publication Types: Review PMID: 15315745 [PubMed - indexed for MEDLINE] 1193: J Biotechnol. 2004 Sep 9;112(3):255-66. Use of quantitative real-time and conventional PCR to assess the stability of the cp4 epsps transgene from Roundup Ready canola in the intestinal, ruminal, and fecal contents of sheep. Alexander TW, Sharma R, Deng MY, Whetsell AJ, Jennings JC, Wang Y, Okine E, Damgaard D, McAllister TA. Agriculture and Agri-Food Canada Research Centre, P.O. Box 3000, Lethbridge, Alta., Canada T1J 4B1. The stability of transgenic DNA encoding the synthetic cp4 epsps protein in a diet containing Roundup Ready (RR) canola meal was determined in duodenal fluid (DF) batch cultures from sheep. A real-time TaqMan PCR assay was designed to quantify the degradation of cp4 epsps DNA during incubation in DF at pH 5 or 7. The copy number of cp4 epsps DNA in the diet declined more rapidly (P < 0.05) in DF at pH 5 as compared to pH 7. The decrease was attributed mainly to microbial activity at pH 7 and perhaps to plant endogenous enzymes at pH 5. The 62-bp fragment of cp4 epsps DNA detected by real-time PCR reached a maximum of approximately 1600 copies in the aqueous phase of DF at pH 7, whereas less than 20 copies were detected during incubations in DF at pH 5. A 1363-bp sequence of cp4 epsps DNA was never detected in the aqueous fraction of DF. Additionally, genomic DNA isolated from RR canola seed was used to test the persistence of fragments of free DNA in DF at pH 3.2, 5, and 7, as well as in ruminal fluid and feces. Primers spanning the cp4 epsps DNA coding region amplified sequences ranging in size from 300 to 1363 bp. Free transgenic DNA was least stable in DF at pH 7 where fragments less than 527 bp were detected for up to 2 min and fragments as large as 1363 bp were detected for 0.5 min. This study shows that digestion of plant material and release of transgenic DNA can occur in the ovine small intestine. However, free DNA is rapidly degraded at neutral pH in DF, thus reducing the likelihood that intact transgenic DNA would be available for absorption through the Peyer's Patches in the distal ileum. Publication Types: Comparative Study Research Support, Non-U.S. Gov't PMID: 15313003 [PubMed - indexed for MEDLINE] 1194: Methods Mol Biol. 2005;286:377-98. Transgenic crops: the current and next generations. Dunwell JM. Department of Agricultural Botany, School of Plant Science, The University of Reading, UK. This chapter describes the present status and future prospects for transgenic (genetically modified) crops. It concentrates on the most recent data obtained from patent databases and field trial applications, as well as the usual scientific literature. By these means, it is possible to obtain a useful perspective into future commercial products and international trends. The various research areas are subdivided on the basis of those associated with input (agronomic) traits and those concerned with output (e.g., food quality) characteristics. Among the former group are new methods of improving stress resistance, and among the latter are many examples of producing pharmaceutical compounds in plants. Publication Types: Review PMID: 15310935 [PubMed - indexed for MEDLINE] 1195: Methods Mol Biol. 2005;286:47-60. Production of hairy root cultures and transgenic plants by Agrobacterium rhizogenes-mediated transformation. Christey MC, Braun RH. New Zealand Institute for Crop & Food Research, Christchurch, New Zealand. Agrobacterium rhizogenes-mediated transformation results in the development of hairy roots at the site of infection. The production of hairy roots involves cocultivation of explants with A. rhizogenes and the subsequent selection of hairy roots on hormone-free medium. Hairy roots have many applications for research including secondary product production and for the study of biochemical pathways. In addition, transgenic plants regenerated from hairy roots often show an altered phenotype due to the presence of the rol genes. In this chapter we describe how to produce and grow hairy root cultures, how to regenerate shoots from these hairy roots, and how to conduct molecular analysis of these cultures. PMID: 15310912 [PubMed - indexed for MEDLINE] 1196: Proc Natl Acad Sci U S A. 2004 Oct 5;101 Suppl 2:14677-82. Epub 2004 Aug 13. Vaccination with genetically engineered allergens prevents progression of allergic disease. Niederberger V, Horak F, Vrtala S, Spitzauer S, Krauth MT, Valent P, Reisinger J, Pelzmann M, Hayek B, Kronqvist M, Gafvelin G, Grönlund H, Purohit A, Suck R, Fiebig H, Cromwell O, Pauli G, van Hage-Hamsten M, Valenta R. Department of Otorhinolaryngology, Vienna General Hospital, University of Vienna, 1090 Vienna, Austria. IgE-mediated allergy affects >25% of the population in industrialized countries. Repeated contact with the disease-eliciting allergens induces rises of allergen-specific IgE Abs and progression of the disease to more severe manifestations. Our study uses a type of vaccine that is based on genetically modified allergen derivatives to treat allergic patients. We developed hypoallergenic derivatives of the major birch pollen allergen, Bet v 1, by genetic engineering and vaccinated birch pollen-allergic patients (n = 124) in a double-blind, placebo-controlled study. Active treatment induced protective IgG Abs that inhibited allergen-induced release of inflammatory mediators. We also observed a reduction of cutaneous sensitivity as well as an improvement of symptoms in actively treated patients. Most important, rises of allergen-specific IgE induced by seasonal birch pollen exposure were significantly reduced in vaccinated patients. Vaccination with genetically engineered allergen derivatives is a therapy for allergy that not only ameliorates allergic reactions but also reduces the IgE production underlying the disease. Publication Types: Clinical Trial Multicenter Study Randomized Controlled Trial Research Support, Non-U.S. Gov't PMID: 15310844 [PubMed - indexed for MEDLINE] 1197: Gastroenterology. 2004 Aug;127(2):502-13. Comment in: Gastroenterology. 2004 Aug;127(2):667-8. Active delivery of trefoil factors by genetically modified Lactococcus lactis prevents and heals acute colitis in mice. Vandenbroucke K, Hans W, Van Huysse J, Neirynck S, Demetter P, Remaut E, Rottiers P, Steidler L. Department for Molecular Biomedical Research, Flanders Interuniversity Institute for Biotechnology and Ghent University, Belgium. BACKGROUND & AIMS: Effective therapeutics for treating acute colitis, caused by disruption of the intestinal epithelial barrier, are scarce. Trefoil factors (TFF) are cytoprotective and promote epithelial wound healing and reconstitution of the gastrointestinal tract, which makes them good candidate therapeutics for acute colitis. However, orally administered TFF stick to the mucus of the small intestine and are absorbed at the cecum. METHODS: We have engineered the food-grade bacterium Lactococcus lactis to secrete bioactive murine TFF. The protective and therapeutic potentials of these TFF-secreting L. lactis were evaluated in parallel with purified TFF in the dextran sodium sulfate (DSS)-induced murine model for acute colitis and in established chronic colitis in interleukin (IL)-10(-/-) mice. Disease was evaluated by blinded macroscopic and microscopic inflammatory scores and by myeloperoxidase activity. RESULTS: Intragastric administration of TFF-secreting L. lactis led to active delivery of TFF at the mucosa of the colon and, in contrast to administration of purified TFF, proved to be very effective in prevention and healing of acute DSS-induced colitis. The in situ secreted murine TFF significantly decreased morbidity and mortality and stimulated prostaglandin-endoperoxide synthase 2 expression, which represents a major therapeutic pathway. In addition, this approach was successful in improving established chronic colitis in IL-10(-/-) mice. CONCLUSIONS: We have positively evaluated a new therapeutic approach for acute and chronic colitis that involves in situ secretion of murine TFF by orally administered L. lactis. This novel approach may lead to effective management of acute and chronic colitis and epithelial damage in humans. Publication Types: In Vitro Research Support, Non-U.S. Gov't PMID: 15300583 [PubMed - indexed for MEDLINE] 1198: Dev Cell. 2004 Aug;7(2):179-92. Comment in: Dev Cell. 2004 Aug;7(2):148-50. Programmed autophagy in the Drosophila fat body is induced by ecdysone through regulation of the PI3K pathway. Rusten TE, Lindmo K, Juhász G, Sass M, Seglen PO, Brech A, Stenmark H. Department of Biochemistry, The Norwegian Radium Hospital, Montebello, N-0310 Oslo. Eukaryotic cells catabolize their own cytoplasm by autophagy in response to amino acid starvation and inductive signals during programmed tissue remodeling and cell death. The Tor and PI3K signaling pathways have been shown to negatively control autophagy in eukaryotes, but the mechanisms that link these effectors to overall animal development and nutritional status in multicellular organisms remain poorly understood. Here, we reveal a complex regulation of programmed and starvation-induced autophagy in the Drosophila fat body. Gain-of-function genetic analysis indicated that ecdysone receptor signaling induces programmed autophagy whereas PI3K signaling represses programmed autophagy. Genetic interaction studies showed that ecdysone signaling downregulates PI3K signaling and that this represents the effector mechanism for induction of programmed autophagy. Hence, these studies link hormonal induction of autophagy to the regulatory function of the PI3K signaling pathway in vivo. Publication Types: Research Support, Non-U.S. Gov't PMID: 15296715 [PubMed - indexed for MEDLINE] 1199: J AOAC Int. 2004 Jul-Aug;87(4):927-36. The modular analytical procedure and validation approach and the units of measurement for genetically modified materials in foods and feeds. Holst-Jensen A, Berdal KG. National Veterinary Institute, Dep., Oslo, Norway. arne.holst-jensen@vetinst.no Food and feed analysts are confronted with a number of common problems, irrespective of the analytical target. The analytical procedure can be described as a series of successive steps: sampling, sample processing, analyte extraction, and ending, finally, in interpretation of an analytical result produced with, e.g., real-time polymerase chain reaction. The final analytical result is dependent on proper method selection and execution and is only valid if valid methods (modules) are used throughout the analytical procedure. The final step is easy to validate-the measurement uncertainty added from this step is relatively limited and can be estimated with a high degree of precision. In contrast, the front-end sampling and processing steps have not evolved much, and the corresponding methods are rarely or never experimentally validated according to internationally harmonized protocols. In this paper, we outlined a strategy for modular validation of the entire analytical procedure, using an upstream validation approach, illustrated with methods for genetically modified materials that may partially apply also to other areas of food and feed analyses. We have also discussed some implications and consequences of this approach in relation to reference materials, measurement units, and thresholds for labelling and enforcement, and for application of the validated methods (modules) in routine food and feed analysis. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 15295887 [PubMed - indexed for MEDLINE] 1200: J Agric Food Chem. 2004 Aug 11;52(16):5223-32. Detection and quantification of roundup ready soy in foods by conventional and real-time polymerase chain reaction. Rott ME, Lawrence TS, Wall EM, Green MJ. Centre for Plant Health, Canadian Food Inspection Agency, 8801 East Saanich Road, Sidney, British Columbia, Canada V8L 1H3. rottm@inspection.gc.ca Transgenic soybean line GTS-40-3-2, marketed under the trade name Roundup Ready (RR) soy, was developed by Monsanto (USA) to allow for the use of glyphosate, the active ingredient of the herbicide Roundup, as a weed control agent. RR soy was first approved in Canada for environmental release and for feed products in 1995 and later for food products in 1996 and is widely grown in Canada. Consumer concern issues have resulted in proposed labeling regulations in Canada for foods derived from genetically engineered crops. One requirement for labeling is the ability to detect and accurately quantify the amount of transgenic material present in foods. Two assays were evaluated. A conventional qualitative Polymerase Chain Reaction (PCR) assay to detect the presence of soy and RR soy and a real-time PCR to quantify the amount of RR soy present in samples that tested positive in the first assay. PCR controls consisted of certified RR soy reference material, single transgenic soybeans, and a processed food sample containing a known amount of RR soy. To test real-world applicability, a number of common grocery store food items that contain soy-based products were tested. For some samples, significant differences in amplification efficiencies during the quantitative PCR assays were observed compared to the controls, resulting in potentially large errors in quantification. A correction factor was used to try to compensate for these differences. PMID: 15291500 [PubMed - indexed for MEDLINE] 1201: J AOAC Int. 2004 May-Jun;87(3):639-46. Suitability of real-time quantitative polymerase chain reaction and enzyme-linked immunosorbent assay for cry9C detection in Mexican corn tortillas: fate of DNA and protein after alkaline cooking. Quirasco M, Schoel B, Plasencia J, Fagan J, Galvez A. Universidad Nacional Autónoma de México, Departamento de Alimentos y Biotecnologia, Facultad de Quimica, 04510, México, D.F., Mexico. Alkaline-cooked corn, called nixtamal, is the basis for many traditional corn products such as tortillas, chips, and taco shells that are used widely in Mexico and Central America and in the preparation of snack foods that are consumed globally. To assess the effects of alkaline and thermal treatments on the detectability of DNA and protein for the presence of genetically modified sequences, various nixtamalized products were prepared from blends of conventional white corn containing 0.1, 1.0, and 10% transgenic corn (event CBH 351, StarLink). Real-time quantitative polymerase chain reactions (RTQ-PCR) and immunoassays were used to determine the cry9C gene and protein, respectively, in unprocessed corn kernels, freshly prepared alkaline-cooked and ground corn (masa), masa flour, tortillas prepared from masa by heat treatment, chips prepared from damp masa dough by deep frying, and from tortillas processed at high (200 degrees C) and low temperatures (70 degrees C). In spite of progressive degradation of genomic DNA during processing, RTQ-PCR genetic analysis allowed detection and quantification of the cry9C gene in all products prepared from 10, 1, and 0.1% StarLink corn, except deep-fried chips containing 0.1% StarLink. Enzyme-linked immunosorbent assays readily detected <1 ppm cry9C protein in all blends of unprocessed corn (10, 1, and 0.1% StarLink) as well as in nonfried tortilla and masa products. This technique was not suitable for thermally treated nixtamalized products containing <1% transgenic corn. Publication Types: Research Support, Non-U.S. Gov't PMID: 15287662 [PubMed - indexed for MEDLINE] 1202: Biotechnol Bioeng. 2004 Aug 20;87(4):495-500. Potato flour viscosity improvement is associated with the expression of a wheat LMW-glutenin gene. Benmoussa M, Vézina LP, Pagé M, Gélinas P, Yelle S, Laberge S. Food Science Department, Purdue University, West Lafayette, Indiana 47907, USA. benmouss@foodsci.purdue.edu It has been previously shown that expression of a high-molecular-weight glutenin (HMW-GS) in transgenic wheat seeds resulted in the improvement of flour functional properties. In this study, potato flour viscosity was improved through a specific expression of a low-molecular-weight glutenin (LMW-GS-MB1) gene in tuber. The resulting construct was introduced into potato leaf explants (Solanum tuberosum cv Kennebec) through Agrobacterium tumefaciens-mediated gene transfer. Southern and Northern analysis of transgenic potato confirmed that the integration of LMW-GS-MB1 in genomic DNA was stable and its mRNA was abundant in transgenic line 16 tubers. Western blot analysis of line 16 extract shows a LMW-GS subunit accumulation in tuber. To demonstrate the capacity of transgenic lines to produce tubers with improved flour functional properties, transgenic lines 9 and 16 exhibiting, respectively, moderate and high expression of LMW-GS-MB1 mRNA and nontransgenic plants were transferred to field plots. The mean viscosity value of flour obtained from the field-grown tubers of transgenic line 16 exhibited a 3-fold increase in viscosity at 23 degrees C when compared to flour from nontransgenic tubers. Publication Types: Research Support, Non-U.S. Gov't PMID: 15286987 [PubMed - indexed for MEDLINE] 1203: Nat Biotechnol. 2004 Aug;22(8):943. Negative fallout from public sentiment in Japan. Watanabe KN, Fujimura T, Shimamoto K, Hashimoto T, Koizumi N, Fukuda H, Naito S, Nakamura K, Mimura T, Ohhashi Y, Shimazaki K, Terashima I, Uchimiya H, Yamaya T. Publication Types: Letter PMID: 15286637 [PubMed - indexed for MEDLINE] 1204: Nahrung. 2004 Jun;48(3):169-76. Influence of cooking and microwave heating on microstructure and mechanical properties of transgenic potatoes. Btaszczak W, Sadowska J, Fornal J, Vacek J, Flis B, Zagórski-Ostoja W. Institute of Animal Reproduction and Food Research, Polish Academy of Sciences, Tuwima 10, PL-10-747 Olsztyn, Poland. viola@pan.olsztyn.pl The transgenic potato clones of cultivar Irga with improved resistance to a necrotic strain of potato virus Y (PVY(N)) were subjected to heat treatment in order to determine their technological quality. The technological quality was determined on the basis of differences between mechanical properties of unmodified potato and transgenic clones during cooking and microwave heating. The compression test was applied in order to evaluate the mechanical resistance of raw, cooked and microwave-treated potatoes. Compression resistance was expressed by fracture stress F (kPa), fracture strain D (mm/mm), and Young modulus E (kPa). The differences in microstructure of potato tubers (unmodified and modified) were investigated using scanning electron microscopy (SEM). The observed differentiation in the mechanical properties of heat-treated potatoes was less connected with genetic modification but most of all with a kind of the process used. The heat processes caused a distinct decrease in mechanical resistance in all the examined tubers. However, the process of microwave heating resulted in more significant changes in mechanical properties of tubers than cooking. Deformation of parenchyma cells during cooking was directly connected with starch, gelatinisation and gel formation. Microwave heating affected significantly cellular water evaporation which resulted in intercellular failure, collapsing of cells, and limitation of starch gelatinisation. Publication Types: Research Support, Non-U.S. Gov't PMID: 15285106 [PubMed - indexed for MEDLINE] 1205: Can J Microbiol. 2004 Jun;50(6):415-21. The fate of a genetically modified Pseudomonas strain and its transgene during the composting of poultry manure. Guan J, Spencer JL, Sampath M, Devenish J. Ottawa Laboratory-Fallowfield, Canadian Food Inspection Agency, Ottawa, Canada. guanj@inspection.gc.ca The fate of the genetically modified (GM) Pseudomonas chlororaphis strain 3732 RN-L11 and its transgene (lacZ insert) during composting of chicken manure was studied using plate count and nested polymerase chain reaction (PCR) methods. The detection sensitivity of the nested PCR method was 165 copies of the modified gene per gram of moist compost or soil. Compost microcosms consisted of a 100-g mixture of chicken manure and peat, whereas soil microcosms were 100-g samples of sandy clay loam. Each microcosm was inoculated with 4 x 1010 CFU of P. chlororaphis RN-L11. In controlled temperature studies, neither P. chlororaphis RN-L11 nor its transgene could be detected in compost microcosms after incubation temperature was elevated to 45 degrees C or above for one or more days. In contrast, in the compost microcosms incubated at 23 degrees C, the target organism was not detected by the plate count method after 6 days, but its transgene was detectable for at least 45 days. In compost bins, the target organism was not recovered from compost microcosms or soil microcosms at different levels in the bins for 29 days. However, the transgene was detected in 8 of the 9 soil microcosms and in only 1 of the 9 compost microcosms. The compost microcosm in which transgene was detected was at the lower level of the bin where temperatures remained below 45 degrees C. The findings indicated that composting of organic wastes could be used to reduce or degrade heat sensitive GM microorganisms and their transgenes. Publication Types: Research Support, Non-U.S. Gov't PMID: 15284887 [PubMed - indexed for MEDLINE] 1206: Int J Food Microbiol. 2004 Sep 1;95(2):127-35. Expression of the catalase gene katA in starter culture Lactobacillus plantarum TISTR850 tolerates oxidative stress and reduces lipid oxidation in fermented meat product. Noonpakdee W, Sitthimonchai S, Panyim S, Lertsiri S. Department of Biochemistry, Faculty of Science, Mahidol University, Bangkok 10400, Thailand. scwnp@mahidol.ac.th The catalase gene katA of Lactobacillus sakei SR911 was cloned and expressed in Escherichia coli UM2 and Lactobacillus plantarum TISTR850 under strong lactococcal promoter P59 in E. coli-lactococcus expression vector pIL1020. The L. plantarum TISTR850 is a catalase-deficient strain isolated from local fermented meat product. The recombinant L. plantarum TISTR850 was shown to decompose hydrogen peroxide, and catalase activity approximately three times higher that of natural catalase-producing strain L. sakei SR911. The recombinant protein was also detected by in situ activity staining of the catalase enzyme. The recombinant L. plantarum TISTR850 did not accumulate hydrogen peroxide under glucose-limited aerobic conditions and remained viable after 60 h of incubation. The recombinant and host strain L. plantarum TISTR850 were used as starter cultures in the fermented meat product, and lipid oxidation was monitored over a 7-day storage at 20 degrees C determined as thiobarbituric acid-reactive substances (TBARS) value. The lipid oxidation level in the fermented meat product seeded with the catalase genetically modified starter culture L. plantarum TISTR850 was significantly lower than that of the natural catalase-deficient strain. PMID: 15282125 [PubMed - indexed for MEDLINE] 1207: J Econ Entomol. 2004 Jun;97(3):871-82. Role of egg density on establishment and plant-to-plant movement by western corn rootworm larvae (Coleoptera: Chrysomelidae). Hibbard BE, Higdon ML, Duran DP, Schweikert YM, Ellersieck MR. USDA-ARS, Plant Genetics Research Unit, 205 Culrtis Hall, University of Missouri, Columbia, MO 65211, USA. The effect of egg density on establishment and dispersal of larvae of the western corn rootworm, Diabrotica virgifera virgifera LeConte, was evaluated in a 3-yr field study. Implications of these data for resistance management plans for Bt crops are discussed. Viable egg levels of 100, 200, 400, 800, and 1600 eggs per infested plant were evaluated in 2000, 2001, and 2002. A 3200 viable egg level was also tested in 2001 and 2002. All eggs were infested on one plant per subplot in a field that was planted to soybean, Glycine max (L.), in the previous year. For each subplot, the infested plant, three plants down the row, the closest plant in the adjacent row of the plot, and a control plant at least 1.5 m from any infested plant (six plants total) were sampled. In 2000, there were five sample dates between egg hatch and pupation, and in 2001 and 2002, there were six sample dates. On each sample date, four replications of each egg density were sampled for both larval recovery and plant damage. Initial establishment on a corn plant seemed to not be density-dependent because a similar percentage of larvae was recovered from all infestation rates. Plant damage and, secondarily, subsequent postestablishment larval movement were density-dependent. Very little damage and postestablishment movement occurred at lower infestation levels, but significant damage and movement occurred at higher infestation rates. Movement generally occurred at a similar time as significant plant damage and not at initial establishment, so timing of movement seemed to be motivated by available food resources rather than crowding. At the highest infestation level in 2001, significant movement three plants down the row and across the 0.76 m row was detected, perhaps impacting refuge strategies for transgenic corn. Publication Types: Research Support, U.S. Gov't, Non-P.H.S. PMID: 15279266 [PubMed - indexed for MEDLINE] 1208: Electrophoresis. 2004 Jul;25(14):2219-26. Sensitive and simultaneous analysis of five transgenic maizes using multiplex polymerase chain reaction, capillary gel electrophoresis, and laser-induced fluorescence. García-Cañas V, González R, Cifuentes A. Institute of Industrial Fermentations (CSIC), Madrid, Spain. The benefits of using multiplex polymerase chain reaction (PCR) followed by capillary gel electrophoresis with laser-induced fluorescence (CGE-LIF) for the simultaneous detection of five transgenic maizes (Bt11, T25, MON810, GA21, and Bt176) are demonstrated. The method uses a hexaplex PCR protocol to amplify the five mentioned transgenic amplicons plus the zein gene used as reference, followed by a CGE-LIF method to analyze the six DNA fragments. CGE-LIF was demonstrated very useful and informative for optimizing multiplex PCR parameters such as time extension, PCR buffer concentration and primers concentration. The method developed is highly sensitive and allows the simultaneous detection in a single run of percentages of transgenic maize as low as 0.054% of Bt11, 0.057% of T25, 0.036% of MON810, 0.064% of GA21, and 0.018% of Bt176 in flour obtaining signals still far from the detection limit (namely, the signal/noise ratios for the corresponding DNA peaks were 41, 124, 98, 250, 252, and 473, respectively). These percentages are well below the minimum threshold marked by the European Regulation for transgenic food labeling (i.e., 0.5-0.9%). A study on the reproducibility of the multiplex PCR-CGE-LIF procedure was also performed. Thus, values of RSD lower than 0.67 and 6.80% were obtained for migration times and corrected peak areas, respectively, for the same sample and three different days (n = 12). On the other hand, the reproducibility of the whole procedure, including four different multiplex PCR amplifications, was determined to be better than 0.66 and 23.3% for migration times and corrected peak areas, respectively. Agarose gel electrophoresis (AGE) and CGE-LIF were compared in terms of resolution and sensitivity for detecting PCR products, demonstrating that CGE-LIF can solve false positives induced by artifacts from the multiplex PCR reaction that could not be addressed by AGE. Moreover, CGE-LIF provides better resolution and sensitivity. To our knowledge, these results demonstrate for the first time that multiplex PCR-CGE-LIF is a solid alternative to determine multiple genetically modified organisms in maize flours in a single run. Publication Types: Research Support, Non-U.S. Gov't PMID: 15274006 [PubMed - indexed for MEDLINE] 1209: Anim Reprod Sci. 2004 Jul;82-83:5-12. Production of bioproducts through the use of transgenic animal models. Keefer CL. Department of Animal & Avian Sciences, University of Maryland, College Park, MD 20742, USA. ckeefer@umd.edu Transgenic livestock that produce recombinant proteins in their milk can provide an economic and safe system for production of valuable proteins, such as pharmaceutical proteins for treatment or prevention of human disease or biomaterials for medical use. This method of production is frequently referred to as biopharming. The promise of biopharming, that is the actual commercial production of pharmaceuticals and other bioproducts, is nearing fulfillment. Improvements in molecular and reproductive techniques and strong economic incentives have continued to drive the implementation of transgenic technology to domestic animals. Nuclear transfer using transgenic donor cells is rapidly becoming the predominant technique used in the production of transgenic livestock, replacing the direct injection of DNA into the zygotic pronuclei. Production of transgenic founder animals by nuclear transfer in combination with traditional reproductive technologies can result in the propagation of transgenic herds of sufficient size to meet market demands for commercially important proteins. While some of the companies that have established transgenic programs have run into setbacks owing to a combination of economic, scientific and regulatory difficulties, other companies are continuing to make significant advances. While further improvements are needed to increase efficiencies of production, economically viable production of recombinant proteins using livestock species is not only possible but should be a commercial reality in the very near future. Publication Types: Review PMID: 15271439 [PubMed - indexed for MEDLINE] 1210: Cloning Stem Cells. 2004;6(2):79-93. Food consumption risks associated with animal clones: what should be investigated? Rudenko L, Matheson JC, Adams AL, Dubbin ES, Greenlees KJ. Center for Veterinary Medicine, Food and Drug Administration, US Department of Health and Human Services, Rockville, Maryland 20855, USA. LRudenko@CVM.FDA.gov Somatic Cell Nuclear Transfer (SCNT), or cloning, is likely to be used for the expansion of elite breeding stock of agronomically important livestock used for food. The Center for Veterinary Medicine at the US Food and Drug Administration has been developing a risk assessment to identify hazards and characterize food consumption risks that may result from cloning. The risk assessment is comprised of two prongs. The first evaluates the health of animal clones, and is referred to as the Critical Biological Systems Approach. The second considers the composition of meat and milk from animal clones. Assessing the safety of food products from animal clones and their progeny, at least during these early stages of the development of the technology, is best accomplished by using both approaches: prospectively drawing on our knowledge of biological systems in development and maturation, and in retrograde, from an analysis of food products. Subtle hazards and potential risks that may be posed by animal clones must, however, be considered in the context of other mutations and epigenetic changes that occur in all food animal populations. PMID: 15268781 [PubMed - indexed for MEDLINE] 1211: Cloning Stem Cells. 2004;6(2):75-207. The Assessment of Food Quality from Cloned Animals. Proceedings of a symposium, November 21-23, 2003, Jouy-en-Josas, France. [No authors listed] Publication Types: Congresses Overall PMID: 15268780 [PubMed - indexed for MEDLINE] 1212: IEEE Eng Med Biol Mag. 2004 Mar-Apr;23(2):52-4. A European perspective on animal cloning and government regulation. Galli C, Duchi R, Lagutina I, Lazzari G. Laboratorio di Tecnologie della Riproduzione, Consorzio per l'Incremento Zootecnico, Associazione Italiana Allevatori, Cremona, Italy. cesare@galli2.191.it PMID: 15264470 [PubMed - indexed for MEDLINE] 1213: Appetite. 2004 Aug;43(1):75-83. Willingness to try new foods as predicted by social representations and attitude and trait scales. Bäckström A, Pirttilä-Backman AM, Tuorila H. Department of Food Technology, P.O. Box 66 (Agnes Sjöbergin katu 2), University of Helsinki, Helsinki FIN-00014, Finland. anna.backstrom@helsinki.fi The structure and predictive ability of social representation of new foods were investigated and compared with instruments measuring relevant attitudes and traits using a questionnaire quantifying these aspects, completed by 743 respondents. Based on their rated willingness to try, new foods were categorized as modified dairy products, genetically modified (GM), organic, and ethnic products (two examples, snails and passion fruit, were treated separately). The social representation (SR) consisted of five dimensions: suspicion of novelties, adherence to technology, adherence to natural food, eating as an enjoyment, and eating as a necessity. The SR dimensions were strong predictors of willingness to try GM foods (predicted by adherence to technology) and organic foods (predicted by adherence to natural foods). Low food neophobia predicted the rated willingness to try snails and passion fruit. Thus, different constructs predicted willingness to try different categories of new foods, and as a whole, SR dimensions markedly improved the prediction. Publication Types: Research Support, Non-U.S. Gov't PMID: 15262020 [PubMed - indexed for MEDLINE] 1214: Appetite. 2004 Aug;43(1):55-64. Predicting intentions to consume functional foods and supplements to offset memory loss using an adaptation of protection motivation theory. Cox DN, Koster A, Russell CG. Commonwealth Scientific Industrial Research Organisation (CSIRO) Health Sciences and Nutrition, P.O. Box 10041, Adelaide, SA 5000, Australia. david.cox@csiro.au The widespread use of dietary supplements and so-called 'functional foods' is thought to be partially motivated by self-control of health. However, whilst consumers want foods associated with well-being or disease prevention, they are unlikely to be willing to compromise on taste or technology. This presents a dilemma for promoters of functional foods. Middle-aged consumers' intentions to consume functional foods or supplements that may improve memory were tested within an adaptation of Protection Motivation theory (PMT). Participants evaluated text descriptions of four products described as: having an unpleasant bitter taste (Natural-FF); having 'additives' to reduce bitterness (Sweetened-FF); being genetically modified to enhance function (GM-FF) and Supplements. Participants were recruited as being of high and low perceived vulnerability to memory failure. In total, 290 middle-aged consumers (aged 40-60 years) participated in the study. Motivations to consume the GM-FF were the lowest. There were gender differences between intention to consume the supplements, Natural-FF and Sweetened-FF and product differences within genders. Women were less favourable than men in their attitudes towards genetic modification in general. Regression analyses indicated that PM predictors of intention to consume functional foods or supplements explained 59-63% of the variance (R2). Overall, perceived 'efficacy' (of the behaviour) and self-efficacy were the most important predictors of intentions to consume. Publication Types: Research Support, Non-U.S. Gov't PMID: 15262018 [PubMed - indexed for MEDLINE] 1215: Genewatch. 2003 Nov-Dec;16(6):12-4. Leveling the field: answers to frequently asked questions about the Cartagena Biosafety Protocol. Freeman L. Harvard Divinity School, USA. PMID: 15255003 [PubMed - indexed for MEDLINE] 1216: Plant Cell Rep. 2004 Nov;23(6):377-85. Epub 2004 Jul 10. Introduction of a citrus blight-associated gene into Carrizo citrange [Citrus sinensis (L.) Osbc. x Poncirus trifoliata (L.) Raf.] by Agrobacterium-mediated transformation. Kayim M, Ceccardi TL, Berretta MJ, Barthe GA, Derrick KS. Institute of Food and Agricultural Sciences, Citrus Research and Education Center, University of Florida, Lake Alfred, FL 33850, USA. seddakum2002@yahoo.com The protein p12 accumulates in leaves of trees with citrus blight (CB), a serious decline of unknown cause. The function of p12 is not known, but sequence analysis indicates it may be related to expansins. In studies to determine the function of p12, sense and antisense constructs were used to make transgenic Carrizo citrange using an Agrobacterium-mediated transformation system. Homogeneous beta-glucuronidase+ (GUS+) sense and antisense transgenic shoots were regenerated using kanamycin as a selective agent. Twenty-five sense and 45 antisense transgenic shoots were in vivo grafted onto Carrizo citrange for further analyses. In addition, 20 sense and 18 antisense shoots were rooted. The homogeneous GUS+ plants contained either the p12 sense or antisense gene (without the intron associated with the gene in untransformed citrus) as shown by PCR and Southern blotting. Northern blots showed the expected RNA in the sense and antisense plants. A protein of identical size and immunoreactivity was observed in seven of nine sense plants but not in nine antisense or non-transgenic plants. At the current stage of growth, there are no visual phenotypic differences between the transgenic and non-transgenic plants. Selected plants will be budded with sweet orange for field evaluation for resistance or susceptibility to CB and general rootstock performance. Publication Types: Research Support, Non-U.S. Gov't PMID: 15248084 [PubMed - indexed for MEDLINE] 1217: Exp Neurol. 2004 Aug;188(2):292-9. Treatment with immunosuppressive and anti-inflammatory agents delays onset of canine genetic narcolepsy and reduces symptom severity. Boehmer LN, Wu MF, John J, Siegel JM. Department of Psychiatry, University of California at Los Angeles, 90095, USA. All Doberman pinschers and Labrador retrievers homozygous for a mutation of the hypocretin (orexin) receptor-2 (hcrtr2) gene develop narcolepsy under normal conditions. Degenerative changes and increased display of major histocompatibility complex class II antigens have been linked to symptom onset in genetically narcoleptic Doberman pinschers. This suggests that the immune system may contribute to neurodegenerative changes and narcoleptic symptomatology in these dogs. We therefore attempted to alter the course of canine genetic narcolepsy, as an initial test of principle, by administering a combination of three immunosuppressive and anti-inflammatory drugs chosen to suppress the immune response globally. Experimental dogs were treated with a combination of methylprednisolone, methotrexate and azathioprine orally starting within 3 weeks after birth, and raised in an environment that minimized pathogen exposure. Symptoms in treated and untreated animals were quantified using the food elicited cataplexy test (FECT), modified FECT and actigraphy. With drug treatment, time to cataplexy onset more than doubled, time spent in cataplexy during tests was reduced by more than 90% and nighttime sleep periods were consolidated. Short-term drug administration to control dogs did not reduce cataplexy symptoms, demonstrating that the drug regimen did not directly affect symptoms. Treatment was stopped at 6 months, after which experimental animals remained less symptomatic than controls until at least 2 years of age. This treatment is the first shown to affect symptom development in animal or human genetic narcolepsy. Our findings show that hcrtr2 mutation is not sufficient for the full symptomatic development of canine genetic narcolepsy and suggest that the immune system may play a role in the development of this disorder. Publication Types: Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S. PMID: 15246829 [PubMed - indexed for MEDLINE] 1218: Trends Biotechnol. 2004 Jul;22(7):331-6. Is confidence in the monitoring of GE foods justified? Heinemann JA, Sparrow AD, Traavik T. New Zealand Institute of Gene Ecology, and School of Biological Sciences, University of Canterbury, Christchurch 8020, New Zealand. jack.heinemann@canterbury.ac.nz Often the limits of detection of genetically engineered organisms (GEOs, LMOs, GMOs) determine what legislation sets as thresholds of allowable contamination of the human food chain with GEOs. Many countries have legislation that is triggered by certain thresholds of contamination. Importantly, international trade in food and animal feed is becoming increasingly vulnerable to interruptions caused by the ambiguity GEOs can create when shipments are monitored at the border. We examine the tools available for detection. Four key error-generating stages are identified with the aim of prompting a higher uniform standard of routine analysis at export and import points. Contamination of the New Zealand corn crop with GEOs is used as a case study for the application of monitoring tools and vulnerability to errors. These tools fail to meet emerging food safety requirements, but some improvements are in development. Publication Types: Comparative Study Evaluation Studies PMID: 15245904 [PubMed - indexed for MEDLINE] 1219: Ann Chim. 2004 Apr;94(4):269-80. Evaluation of purification procedures of DNA from maize-meal samples by exploiting different analytical techniques for the assessment of DNA quality. Vanni A, Anfossi L, Giovannoli C, Oddenino L, Giraudi G. Department of Analytical Chemistry, University of Turin, Via Giuria 5, 10125 Torino, Italy. adriano.vanni@unito.it Two different approaches generally applied to achieve purification of DNA extracted from cells were compared: precipitation by organic solvents and enzymatic treatments. We investigated various experimental protocols reported in literature by evaluating DNA purity, integrity and yield. Reliability of analytical techniques normally employed to assess DNA purity and quantity was studied and comments and conclusions were suggested by comparing results obtained by different analytical techniques. Enzymatic treatments prove to be unable of increasing DNA purity while determining a significant degradation. In contrast, optimised conditions for solvent precipitation enabled a sharp increase of DNA purity to be obtained, associated with the maintenance of the initial DNA integrity. The application of the optimised protocol to maize-meal samples allowed us to achieve a good PCR amplification even with those samples which gave poor amplification by following the protocol recommended by the Italian legislation in force for GMO detection in food. Publication Types: Research Support, Non-U.S. Gov't PMID: 15242092 [PubMed - indexed for MEDLINE] 1220: J Agric Food Chem. 2004 Jul 14;52(14):4535-40. Detection of genetically modified soybean using peptide nucleic acids (PNAs) and microarray technology. Germini A, Mezzelani A, Lesignoli F, Corradini R, Marchelli R, Bordoni R, Consolandi C, De Bellis G. Dipartimento di Chimica Organica e Industriale, Università degli Studi di Parma, Parco Area delle Scienze 17/A, 43100 Parma, Italy. Peptide nucleic acid (PNA) microarrays for the detection of Roundup Ready soybeans in food have been prepared. PNA probes are known to be more efficient and selective in binding DNA sequences than the analogous oligonucleotides and are very suitable to be used for diagnostics in food. PNAs of different lengths were carefully designed and synthesized by solid-phase synthesis on an automatic synthesizer adopting the BOC strategy. PNAs were purified by HPLC and characterized by HPLC/MS. The probes were spotted on a functionalized surface to produce a microarray to be hybridized with PCR products. DNA extracted from reference material was amplified using Cy3- and Cy5-labeled primers, and the fluorescent PCR products obtained were hybridized on the microarray. Two protocols were adopted: the hybridization with dsDNA or with ssDNA obtained by digestion with the enzyme lambda exonuclease. The best results were obtained using a 15-mer PNA probe in combination with the ssPCR product derived from enzymatic digestion. The method was applied to the analysis of a sample of certified transgenic soybean flour. PMID: 15237963 [PubMed - indexed for MEDLINE] 1221: World J Gastroenterol. 2004 Jul 15;10(14):2063-6. Effects of fucosylated milk of goat and mouse on Helicobacter pylori binding to Lewis b antigen. Xu HT, Zhao YF, Lian ZX, Fan BL, Zhao ZH, Yu SY, Dai YP, Wang LL, Niu HL, Li N, Hammarström L, Borén T, Sjöström R. State Key Laboratories for Agrobiotechnology, China Agriculture University, Beijing 100094, China. AIM: To evaluate the effects of animal milk containing fucosylated antigens on Helicobacter pylori (H. pylori) binding to Lewis b antigen. METHODS: A mammary gland expression vector containing human alpha1-3/4-fucosyltransferase cDNA sequences was constructed. Transient expression of human alpha1-3/4-fucosyltransferase cDNA in goat mammary cell and establishment of transgenic mice were performed. The adhesion inhibitory properties of milk samples were analyzed by using H. pylori. RESULTS: Goat milk samples were found to inhibit bacterial binding to Lewis b antigen. The highest inhibition was observed 42 h after injection of the plasmid. The binding activity of H. pylori to Lewis b antigen reduced mostly, by 83%, however milk samples from transgenic mice did not inhibit H. pylori binding to Lewis b antigen. CONCLUSION: The use of "humanized" animal milk produced by the transgenic introduction of fucosylated antigen can perhaps provide an alternative therapy and preventive measure for H. pylori infection. PMID: 15237435 [PubMed - indexed for MEDLINE] 1222: Physiol Behav. 2004 Jul;81(5):741-8. Is dopamine required for natural reward? Cannon CM, Bseikri MR. Department of Biochemistry and Howard Hughes Medical Institute, University of Washington, Box 357370, Seattle, WA 98195-7370, USA. caesia@u.washington.edu Reward is fundamental to the organization of behavior, and the neurotransmitter dopamine (DA) is widely recognized to be critical to the neurobiology of reward, learning and addiction. Virtually all drugs of abuse, including heroin and other opiates, alcohol, cocaine, amphetamine and nicotine activate dopaminergic systems. So called "natural" rewards such as food, positive social interactions and even humor, likewise activate DA neurons and are powerful aids to attention and learning. Sweet solutions are a well-characterized natural reward. When a source of sugar is encountered, animals will consume substantial amounts, return to it preferentially, and will work to obtain access. Dopamine systems are activated in animals drinking sugar solutions, and lesions of dopaminergic neurons or pharmacological blockade of DA receptors seem to reduce the reward value of both sweet tastes and drugs of abuse. However, we have recently demonstrated that genetically modified mice that cannot make DA (DD mice) manifest normal sucrose preference. During preference tests, mutant mice initiated licking less frequently than did normal mice, but the rate of licking by DD mice for sweets was actually higher than that of normal mice, indicating that their motor ability to lick is intact. We conclude that DA is not required for the hedonic response to sweets nor for their discrimination. This brief and slightly humorous review discusses these findings in the context of current and historical answers to the question, "What is the role of DA in reward?" Publication Types: Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. Review PMID: 15234179 [PubMed - indexed for MEDLINE] 1223: Mol Ther. 2004 Jul;10(1):1. Agbiotech: success depends on trust. Verma IM. Publication Types: Editorial PMID: 15233936 [PubMed - indexed for MEDLINE] 1224: Nat Biotechnol. 2004 Jul;22(7):803. Comment on: Nat Biotechnol. 2004 Feb;22(2):133. Drugs in Mexican crops? Acevedo F. Publication Types: Comment Letter PMID: 15229533 [PubMed - indexed for MEDLINE] 1225: Nat Biotechnol. 2004 Jul;22(7):803. Comment on: Nat Biotechnol. 2004 Feb;22(2):133. The facts on flax. Cummins J. Publication Types: Comment Letter PMID: 15229532 [PubMed - indexed for MEDLINE] 1226: Nat Biotechnol. 2004 Jul;22(7):791. Agbiotech firms realign product focus. Fox JL. Publication Types: News PMID: 15229525 [PubMed - indexed for MEDLINE] 1227: J Anim Sci. 2004 Jun;82(6):1693-8. Nutritional value of a corn containing a glutamate dehydrogenase gene for growing pigs. Guthrie TA, Apgar GA, Griswold KE, Lindemann MD, Radcliffe JS, Jacobson BN. Department of Animal Science, Food, and Nutrition, Southern Illinois University, Carbondale 62901, USA. Eight female PIC Line 42 pigs (initial BW = 47.5 +/- 1.8 kg) were used in a two-period switchback design (n = 4 per treatment per period) to evaluate the nutritional difference between a genetically modified corn and a similar nontransgenic corn. The genetically altered corn (gdhA+) contained a glutamate dehydrogenase gene isolated from Escherichia coli. The non-transgenic corn was the same variety lacking the transgenic cassette, grown at the same two locations. Pigs were surgically fitted with steered ileocecal valve cannulas for collection of ileal digesta. Diets were made up of primarily one of the two corn sources. Dietary AA profiles were adjusted using crystalline AA to match Illinois Ideal Protein Ratios. Pigs were limit-fed at 8% of metabolic body weight (BW0.75) in two equal feedings at 0600 and 1800 daily throughout the experiment. The study consisted of two 15-d periods. Each period consisted of a 7-d acclimation period, a 3-d total collection of feces and urine, two 12-h ileal collections, and a 3-d adjustment period between ileal collections to ensure adequate hydration. Crude protein, leucine, methionine, alanine, aspartic acid, glutamic acid, and tyrosine concentrations were greater (P < 0.05) in the gdhA+ corn than in the nontransgenic variety. The presence of the gene did not alter (P > 0.17) BW gain. Similarly, DM digestibility, fecal N excretion (grams per day), apparent total-tract N digestibility, N balance, net protein utilization, and N retained as percentages of absorbed were not affected (P > or = 0.32) by the gene modification. Apparent ileal AA digestibility values did not differ (P > 0.31) between the two dietary treatments. Results of this study suggest corn that contains the E coli. gene for glutamate dehydrogenase was nutritionally equivalent to the unaltered variety. Publication Types: Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. PMID: 15216996 [PubMed - indexed for MEDLINE] 1228: Nature. 2004 Jun 24;429(6994):826. Plant biochemistry: a naturally decaffeinated arabica coffee. Silvarolla MB, Mazzafera P, Fazuoli LC. Centro de Análise e Pesquisa Tecnológica do Agronegócio do Café Alcides Carvalho, IAC-APTA, CP 28, CEP 13001-970, Campinas, SP, Brasil. The adverse side effects of caffeine have increased the market for decaffeinated coffee to about 10% of coffee consumption worldwide (http://www.ncausa.org), despite the loss of key flavour compounds in the industrial decaffeinating process. We have discovered a naturally decaffeinated Coffea arabica plant from Ethiopia, a species normally recognized for the high quality of its beans. It should be possible to transfer this trait to commercial varieties of arabica coffee plants by intraspecific hybridization--a process likely to be simpler than an interspecific hybridization strategy, which could require more than 30 years of breeding to fix the decaffeinated trait and would probably result in an inferior cup of coffee. PMID: 15215853 [PubMed - indexed for MEDLINE] 1229: Anal Bioanal Chem. 2004 Mar;378(6):1616-23. Analysis and interpretation of data from real-time PCR trace detection methods using quantitation of GM soya as a model system. Burns MJ, Valdivia H, Harris N. BioAnalytical Innovation Team, LGC Limited, Queens Road, Teddington, Middlesex, TW11 OLY, UK. Malcolm.Burns@lgc.co.uk Recent years have seen an increased interest in DNA trace detection methods involved in many areas of bioanalytical research, such as quantitation of genetically modified (GM) ingredients in food products. There is little in the way of standardisation of data handling from these methods, and the data generated needs to be analysed appropriately if the results are to be interpreted correctly. This paper describes particular aspects of real-time PCR trace detection methods in order to increase the understanding of data generated using this bioanalytical technique. Using the specific example of GM soya detection and quantitation, it focuses on the production of calibration curves based on the mean and individual data values, the interpretation of correlation coefficients, regression techniques, and discusses suitable data analysis arising from simple and more complex experimental designs following transformation. By using the approaches outlined in this paper, more accurate analysis of data from real-time PCR and GM trace detection methods could be achieved. Publication Types: Research Support, Non-U.S. Gov't PMID: 15214425 [PubMed - indexed for MEDLINE] 1230: Am J Clin Nutr. 2004 Jul;80(1):131-6. Lutein and beta-carotene from lutein-containing yellow carrots are bioavailable in humans. Molldrem KL, Li J, Simon PW, Tanumihardjo SA. Integrated Graduate Program in Nutritional Sciences, University of Wisconsin, Madison, 53706, USA. BACKGROUND: Lutein is a hydroxy-carotenoid constituting the macular pigment of the human retina. Increasing lutein intake from foods could increase the density of this pigment and decrease the risk of developing macular degeneration. Yellow carrots are a novel food source that could increase lutein consumption. OBJECTIVE: We evaluated and compared lutein uptake and clearance in humans from genetically selected lutein-containing yellow carrots fed chronically and from a lutein supplement. DESIGN: Four women and 5 men aged 23-28 y participated in this randomized, blinded, 3 x 3 crossover intervention. Treatments consisted of yellow carrots (YC treatment, 1.7 mg lutein/d), white carrots as a negative control (WC treatment, 0 mg lutein/d), and a lutein supplement in oil as a positive control (LS treatment, 1.7 mg lutein/d). Each treatment lasted 7 d and was followed by a 7-d washout period. RESULTS: Mean (+/- SD) peak changes in serum lutein concentration from baseline were 0.31 +/- 0.08, 0.19 +/- 0.08, and -0.04 +/- 0.04 micromol/L for the LS, YC, and WC treatments, respectively. The areas under the curve for 0-14 d (AUC(0-14d)) differed significantly (P < 0.0001) between treatments. Lutein from the YC treatment was 65% as bioavailable as that from the LS treatment. The AUC(0-14d) for beta-carotene (-0.01 +/- 0.28 micromol.d/L) also showed that the YC treatment maintained peak serum beta-carotene concentrations at 0.35 +/- 0.30 micromol/L, whereas the LS treatment did not (AUC(0-14d) = -0.71 +/- 0.59 micromol.d/L). CONCLUSION: Lutein from this novel food source significantly increases serum lutein concentrations and does not result in the decrease in beta-carotene concentrations that accompanies administration of lutein supplements. Publication Types: Clinical Trial Randomized Controlled Trial Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. PMID: 15213039 [PubMed - indexed for MEDLINE] 1231: J Agric Food Chem. 2004 Jun 30;52(13):4149-58. Composition of grain and forage from corn rootworm-protected corn event MON 863 is equivalent to that of conventional corn (Zea mays l.). George C, Ridley WP, Obert JC, Nemeth MA, Breeze ML, Astwood JD. Product Safety Center, Monsanto Company, 800 North Lindbergh Boulevard, St Louis, Missouri 63167, USA. cherian.george@monsanto.com Insect-protected corn hybrids containing event MON 863 protect corn plants against feeding damage from corn rootworm (Diabrotica), a major North American insect pest. Corn event MON 863 contains a gene that expresses an amino acid sequence variant of the wild-type Cry3Bb1 insecticidal protein from Bacillus thuringiensis. The purpose of this study was to compare the composition of corn containing event MON 863 with that of conventional nontransgenic corn. Compositional analyses were conducted to measure proximates, fiber, amino acids, fatty acids, minerals, folic acid, thiamin, riboflavin, vitamin E, antinutrients, and certain secondary metabolites in grain and proximates and fiber content in forage collected from a total of eight field sites in the U.S. and Argentina. Compositional analyses demonstrated that the grain and forage of event MON 863 are comparable in their nutritional content to the control corn hybrid and conventional corn. These comparisons, together with the history of the safe use of corn as a common component of animal feed and human food, support the conclusion that corn event MON 863 is compositionally equivalent to, and as safe and nutritious as, conventional corn hybrids grown commercially today. PMID: 15212462 [PubMed - indexed for MEDLINE] 1232: Environ Sci Technol. 2004 May 15;38(10):174A-175A. Ecologists call for caution with engineered organisms. Pelley J. Publication Types: News PMID: 15212238 [PubMed - indexed for MEDLINE] 1233: Int J Occup Med Environ Health. 2004;17(1):47-57. The Precautionary Principle: implications for risk management strategies. Saltelli A, Funtowicz S. European Commission, Joint Research Centre, Institute for the Protection and Security of the Citizen, Ispra, VA, Italy. andrea.saltelli@jrc.it The European Commission has published a Communication on the Precautionary Principle and a White Book on Governance. These provide us (as research civil servants of the Commission) an institutional framework for handling scientific information that is often incomplete, uncertain, and contested. But, although the Precautionary Principle is intuitively straightforward to understand, there is no agreed way of applying it to real decision-making. To meet this perceived need, researchers have proposed a vast number of taxonomies. These include ignorance auditing, type one-two-three errors, a combination of uncertainty and decision stakes through post-normal science and the plotting of ignorance of probabilities against ignorance of consequences. Any of these could be used to define a precautionary principle region inside a multidimensional space and to position an issue within that region. The role of anticipatory research is clearly critical but scientific input is only part of the picture. It is difficult to imagine an issue where the application of the Precautionary Principle would be non-contentious. From genetically-modified food to electro-smog, from climate change to hormone growth in meat, it is clear that: 1) risk and cost-benefit are only part of the picture; 2) there are ethical issues involved; 3) there is a plurality of interests and perspectives that are often in conflict; 4) there will be losers and winners whatever decision is made. Operationalization of the Precautionary Principle must preserve transparency. Only in this way will the incommensurable costs and benefits associated with different stakeholders be registered. A typical decision will include the following sorts of considerations: 1) the commercial interests of companies and the communities that depend on them; 2) the worldviews of those who might want a greener, less consumerist society and/or who believe in the sanctity of human or animal life; 3) potential benefits such as enabling the world's poor to improve farming; 4) risks such as pollution, gene-flow, or the effects of climate change. In this paper we will discuss the use of a combination of methods on which we have worked and that we consider useful to frame the debate and facilitate the dialogue among stakeholders on where and how to apply the Precautionary Principle. Publication Types: Comparative Study Review PMID: 15212206 [PubMed - indexed for MEDLINE] 1234: Wei Sheng Yan Jiu. 2004 May;33(3):303-6. [Nutrition assessment of transgenic rice] [Article in Chinese] Li Y, Piao J, Chen X, Zhuo Q. National Institute for Nutrition and Food Safety, Chinese Center for Disease Control and Prevention, Beijing 100050, China. OBJECTIVE: To observe the nutrition effects between transgenic rice and non-transgenic rice. METHODS: Wistar rats were divided into three groups according to their sex respectively: transgenic rice group, non-transgenic rice group and AIN93G normal control group. They were fed with corresponding food for 28 days. Indicator as follows: food intake, protein effective ratio, body weight, body length, blood routine test, blood biochemistry test, organ index and bone density. RESULTS: In male groups, expect that liver/body ratio was higher in transgenic rice group than that in non-transgenic rice group, all other indicators had no significant difference each other. In female groups, liver/body weight ratio, blood calcium and bone density were higher in transgenic group than those in non-transgenic group. All other indicators had no significant difference between transgenic rice group and non-transgenic rice group. Body weight, bone density and blood calcium were higher and liver/body weight ratio had no significant difference in transgenic rice group than those in AIN93G group. CONCLUSION: Transgenic rice had good nutrition effects on rat development and no toxicity and adverse effects were found in transgenic rice group. Publication Types: English Abstract Research Support, Non-U.S. Gov't PMID: 15211798 [PubMed - indexed for MEDLINE] 1235: Cell. 2004 Jun 25;117(7):981-91. Taste representations in the Drosophila brain. Wang Z, Singhvi A, Kong P, Scott K. Department of Molecular and Cell Biology and Helen Wills Neuroscience Institute, 291 Life Sciences Addition, University of California, Berkeley, Berkeley, California 94720, USA. Drosophila taste compounds with gustatory neurons on many parts of the body, suggesting that a fly detects both the location and quality of a food source. For example, activation of taste neurons on the legs causes proboscis extension or retraction, whereas activation of proboscis taste neurons causes food ingestion or rejection. We examined whether the features of taste location and taste quality are mapped in the fly brain using molecular, genetic, and behavioral approaches. We find that projections are segregated by the category of tastes that they recognize: neurons that recognize sugars project to a region different from those recognizing noxious substances. Transgenic axon labeling experiments also demonstrate that gustatory projections are segregated based on their location in the periphery. These studies reveal the gustatory map in the first relay of the fly brain and demonstrate that taste quality and position are represented in anatomical projection patterns. Publication Types: Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. PMID: 15210117 [PubMed - indexed for MEDLINE] 1236: Risk Anal. 2004 Jun;24(3):727-35. Using risk assessment, benefit-cost analysis, and real options to implement a precautionary principle. Farrow S. scott.farrow@verizon.net Risk assessment is an established methodology for environmental and public health issues. However, economists' core approach to both risk assessment and risk management, benefit-cost analysis, often fails to transparently evaluate variability in a way that is a trademark of quantitative risk assessment. Concurrently, environmental advocates are proposing new management criteria based on a vaguely framed "Precautionary Principle." This manuscript demonstrates how risk assessment techniques for characterizing variability, benefit-cost analysis, and decision-making criteria under uncertainty and irreversibility can be combined. The result is a quantifiable, case-specific, and risk-dependent "precautionary" threshold for action compared to standard benefit-cost approaches. The Clean Air Act and the regulation of genetically modified corn provide applications. Publication Types: Research Support, Non-U.S. Gov't PMID: 15209941 [PubMed - indexed for MEDLINE] 1237: Risk Anal. 2004 Jun;24(3):715-26. Explaining public resistance to genetically modified corn: an analysis of the distribution of benefits and risks. Wu F. Environmental and Occupational Health, University of Pittsburgh, Pittsburgh, PA 15261, USA. Genetically modified (GM) crops have met with widespread approval among scientists and policy makers in the United States, but public approval of GM crops, both domestically and abroad, is progressing much more slowly. An underlying cause of public wariness may be that both nations and individual consumers do not perceive significant benefits to themselves from GM crops, while fearing the risks they may incur. In this study, an economic analysis is conducted to determine whether the benefits of one type of GM corn, Bt corn (genetically modified to resist damage from the ECB and Southwestern corn borer), outweigh the potential risks; and who the "winners" and "losers" are among stakeholder groups that may be affected by Bt corn. It is found that Bt corn growers, consumers, and industry all benefit from Bt corn adoption, though the purported health and environmental benefits of reducing chemical pesticide usage through Bt corn are negligible. Though the aggregated public benefit is large, the welfare gain to individual consumers is small and may not make up for perceived risks. While environmental and health risks of Bt corn are unlikely, the potential market risks-impacting both the organic corn market and total U.S. corn exports-are found to be significant. Currently, distributional analysis is not a part of regulatory decision making of Bt corn in the United States; yet it may help to explain why decision makers at both the government and individual-consumer levels have failed to embrace Bt corn and other GM crops. Publication Types: Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. PMID: 15209940 [PubMed - indexed for MEDLINE] 1238: Clin Lab. 2004;50(5-6):380-1. Consumer protection from an EU regulation on the mandatory labelling of genetically modified food. [No authors listed] On the 7. November 2003 a new regulation was enforced in the states of the EU to govern the authorisation and labelling of genetically modified food in standardized and legally binding form. Raw materials from GM crops now have to feature in the list of ingredients of the end products. The consumer is free to choose whether or not he accepts gene technology in his food purchases. PMID: 15209444 [PubMed - indexed for MEDLINE] 1239: Wei Sheng Yan Jiu. 2004 Mar;33(2):233-6. [Safety evaluation of food from transgenic fish and the molecular biological mechanism] [Article in Chinese] Zhang X, Yang X. Institute of Nutrition and Food Safety, Chinese Center for Disease Control and Prevention, Beijing 100050, China. More progresses have been made in the studying of transgenic fish in China, but the studying work of safety evaluation of food from transgenic fish are started up just now. Compared to plants and animals on the land, it is more difficult to control the mobility of fish and fish can give birth to a large number of offsprings, so the ecological risk or hazard about transgenic fish is more critical than others. Another safety problem is the chimerism which is initiated by the gene transfer methods used in the transgenic fish. Getting sterile triploid transgenic fish and fixed point integration are efficient to solve the two problems above respectively. The solution of the two problems are also the basis of safety evaluation and detection of food from transgenic fish. Up to now, there are little reports on the safety evaluation of transgenic fish including nutritional evaluation and allergic reaction, and there are no basic research on the detection of transgenic fish for the aim of food safety. In brief, it is very urgent to start up the research on the safety evaluation and detection of transgenic fish for the control of food safety. Publication Types: English Abstract Research Support, Non-U.S. Gov't Review PMID: 15209016 [PubMed - indexed for MEDLINE] 1240: Wei Sheng Yan Jiu. 2004 Mar;33(2):176-9. [Study on food safety of genetically modified rice which expressed cowpea trypsin inhibitor by 90 day feeding test on rats] [Article in Chinese] Zhuo Q, Chen X, Piao J, Gu L. Institute of Nutrition and Food Safety, Chinese Center for Disease Control and Prevention, Beijing 100050, China. OBJECTIVE: Rats were fed by transgenic rice which expressed insecticidal protein CPTI (cowpea trypsin inhibitor) to study if the transgenic rice possessed potential toxic or adverse effects. METHODS: Weanling Wistar rats were randomly divided into three groups: T, N and C group. The diet of T group contained 78.3% transgenic rice. The diet of N group contained 74.7% non transgenic rice which was the parent line of the transgenic one. The diet formula of C group was AIN93G. The macro- and micronutrient content were equal in three diets. The rats were fed for 90 days. Food intakes were weight every day, body-weight were weight and body-length were measured every week. In the middle and at the end of feeding period, haematological value and clinical chemistry parameters were measured, at the end of the 90th day, post-mortem organ coefficient were measured, organ tissues analysis was performed and bone density was measured. RESULTS: In most situation, there were no significant differences among the three groups(P > 0.05) and no histopathological damage were detected. At the end of the 1st month, the male rats' body length of the T group was longer than the other two groups and at the end of the test period, the male rats' blood glucose and ALT were lower than the other two groups. In the middle of the test period, the female rats' red blood cell number and hemoglobin were higher than the other two groups and at the end of the test period, the female rats' monocyte number was higher than the other two groups (P < 0.05). But all of the results were in the normal range which had been reported before. CONCLUSION: From the results of the 90 days feeding test of transgenic rice on rats there did not reveal any signs of toxic and adverse effects. Publication Types: English Abstract Research Support, Non-U.S. Gov't PMID: 15208998 [PubMed - indexed for MEDLINE] 1241: Plant Physiol. 2004 Jun;135(2):709-14. Fluorescent screening of transgenic Arabidopsis seeds without germination. Wei S, Bravdo BA, Shoseyov O. The Otto Warburg Minerva Center for Agricultural Biotechnology, Faculty of Agricultural, Food and Environmental Quality Sciences, The Hebrew University of Jerusalem, Rehovot 76100, Israel. In this paper, we describe a reliable method for the screening and selection of Arabidopsis transgenic seeds within minutes without germination. Expression of the Aspergillus niger beta-glucosidase gene BGL1 in the plant's endoplasmic reticulum was used as a visual marker, together with 4-methylumbelliferyl-beta-D-glucopyranoside (MUGluc) as a substrate. Subsequent to incubation in a solution of MUGluc at room temperature for 2 to 15 min, transgenic seeds expressing BGL1 demonstrated a distinct fluorescent signal under UV light. Optimal screening conditions at room temperature were achieved between 75 and 450 microm MUGluc, at a pH of 2.5 to 5.0 and 2 to 5 min of incubation. No significant loss of viability was detected in transgenic seeds that were redried and stored for 45 d after incubation in MUGluc solution for 2 to 150 min. Transgenic plants expressing BGL1 displayed normal phenotypes relative to the wild type. Selection frequency was 3.1% +/- 0.34% for the fluorescence selection method, while kanamycin resistant selection resulted in only 0.56% +/- 0.13% using the same seed batch. This novel selection method is nondestructive, practical, and efficient, and eliminates the use of antibiotic genes. In addition, the procedure shortens the selection time from weeks to minutes. Publication Types: Research Support, Non-U.S. Gov't PMID: 15208418 [PubMed - indexed for MEDLINE] 1242: J Exp Bot. 2004 Jul;55(402):1445-54. Epub 2004 Jun 18. Real-time PCR: what relevance to plant studies? Gachon C, Mingam A, Charrier B. Institut de Biotechnologie des Plantes, UMR CNRS 8618, Université Paris-Sud, F-91405 Orsay cedex, France. The appearance of genetically modified organisms on the food market a few years ago, and the demand for more precise and reliable techniques to detect foreign (transgenic or pathogenic) DNA in edible plants, have been the driving force for the introduction of real-time PCR techniques in plant research. This was followed by numerous fundamental research applications aiming to study the expression profiles of endogenous genes and multigene families. Since then, the interest in this technique in the plant scientist community has increased exponentially. This review describes the technical features of quantitative real-time PCR that are especially relevant to plant research, and summarizes its present and future applications. Publication Types: Review PMID: 15208338 [PubMed - indexed for MEDLINE] 1243: Chem Res Toxicol. 2004 Jun;17(6):814-8. Mutations induced by the carcinogenic pyrrolizidine alkaloid riddelliine in the liver cII gene of transgenic big blue rats. Mei N, Heflich RH, Chou MW, Chen T. Division of Genetic and Reproductive Toxicology, National Center for Toxicological Research, Food and Drug Administration, Jefferson, Arkansas 72079, USA. nmei@nctr.fda.gov Riddelliine is a naturally occurring pyrrolizidine alkaloid that forms a number of different mononucleotide and dinucleotide adducts in DNA. It is a rodent carcinogen and a potential human hazard via food contamination. To examine the mutagenicity of riddelliine, groups of six female transgenic Big Blue rats were gavaged with 0.1, 0.3, and 1.0 mg riddelliine per kg body weight. The middle and high doses resulted in liver tumors in a previous carcinogenesis bioassay. The animals were treated 5 days a week for 12 weeks and sacrificed 1 day after the last treatment. The liver DNA was isolated for analysis of the mutant frequency (MF) in the transgenic cII gene, and the types of mutations were characterized by sequencing the mutants. A significant dose-dependent increase in MF was found, increasing from 30 x 10(-)(6) in the control animals to 47, 55, and 103 x 10(-)(6) in the low, middle, and high dose groups, respectively. Molecular analysis of the mutants indicated that there was a statistically significant difference between the mutational spectra from the riddelliine-treated and the control rats. A G:C --> T:A transversion (35%) was the major type of mutation in rats treated with riddelliine, whereas a G:C --> A:T transition (55%) was the predominant mutation in the controls. In addition, mutations from the riddelliine-treated rats included an unusually high frequency (8%) of tandem base substitutions of GG --> TT and GG --> AT. These results indicate that riddelliine is a genotoxic carcinogen in rat liver and that the types of mutations induced by riddelliine are consistent with riddelliine adducts involving G:C base pairs. Publication Types: Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S. PMID: 15206902 [PubMed - indexed for MEDLINE] 1244: Int J Toxicol. 2004 Mar-Apr;23(2):79-80. Genetically modified foods get bad rap. Mehendale HM. Publication Types: Editorial PMID: 15204724 [PubMed - indexed for MEDLINE] 1245: J Drug Target. 2003;11(8-10):539-45. Advantageous features of plant-based systems for the development of HIV vaccines. Horn ME, Pappu KM, Bailey MR, Clough RC, Barker M, Jilka JM, Howard JA, Streatfield SJ. ProdiGene, College Station, TX 77845, USA. Plants have recently become an attractive option for the production of recombinant proteins. Plant-based systems can be used to produce many classes of foreign proteins including candidate vaccine antigens. The selected antigen can be purified from plant material prior to delivery by the preferred route, or alternatively delivered orally in edible plant material that has been processed to give a homogeneous and stable product. Several plant species have been used to express a wide range of vaccine candidates with tobacco, potato and corn being particularly favored. Corn seed is especially well suited to various food processing technologies that generate dry homogeneous material suitable for extended storage and refrigeration-free transport and distribution. Many antigens have been expressed in corn and assessed for efficacy in trials with generally positive results. Candidate HIV vaccines are particularly good targets for plant-based oral delivery since there is a great need for an easily distributed affordable vaccine that could be administered without injection and induce strong mucosal immune responses. As a first step in evaluating plant expression technology with a relevant antigen that might easily be tested in an animal system, we expressed the SIV major surface glycoprotein gp130 (analogous to HIV gp120) in corn seed. Expression levels were achieved that are compatible with conducting oral delivery trials in animals. Publication Types: Review PMID: 15203923 [PubMed - indexed for MEDLINE] 1246: Transgenic Res. 2004 Apr;13(2):143-54. Bi-directional duplex promoters with duplicated enhancers significantly increase transgene expression in grape and tobacco. Li ZT, Jayasankar S, Gray DJ. Mid-Florida Research and Education Center, Institute of Food and Agricultural Sciences, University of Florida, 2725 Binion Road, Apopka, FL 32703-8504, USA. Novel bi-directional duplex promoters (BDDP) were constructed by placing two identical core promoters divergently on both upstream and downstream sides of their duplicated enhancer elements. Estimates of promoter function were obtained by creating versions of CaMV 35S and CsVMV BDDPs that contained reporter marker genes encoding beta-glucuronidase (GUS) and enhanced green fluorescent protein (EGFP) interchangeably linked either to the upstream or downstream core promoters. GUS was used for quantitative analysis of promoter function, whereas, EGFP allowed visual qualitative evaluation. In addition, the GUS and EGFP genes placed in downstream positions were modified by translational fusion with neomycin phosphotransferase (NPTII) to allow simultaneous monitoring of promoter activity and selection of stable transformants. These versions of BDDP were compared with each other and with equivalent unidirectional constructs by evaluating their expression in grape and tobacco. For 35S promoter constructs tested in grape somatic embryos (SE), BDDP exhibited transient GUS expression 206- and 300-fold greater in downstream and upstream configurations, respectively, compared to a unidirectional 35S core promoter. Compared with a unidirectional double enhanced 35S promoter, BDDPs exhibited 0.5- and 3-fold increased GUS expression from downstream and upstream core promoters, respectively. The same differences in expression levels determined quantitatively with GUS were distinguished qualitatively with EGFP. Constructs using CsVMV core promoters yielded results relative to those obtained with 35S promoter. For example, the upstream BDDP CsVMV core promoter provided a 200-fold increase in GUS expression compared to a unidirectional core promoter. However, CsVMV promoter was found to have higher promoter activity than 35S promoter in both BDDP and unidirectional constructs. Incorporation of an additional duplicated enhancer element to BDDPs resulted in increased expression. For example, a 35S BDDP with two divergently arranged duplicated enhancer elements resulted in over a 6-fold increase in GUS expression in stably transformed tobacco plants compared to a BDDP with one duplicated enhancer element. Data demonstrate that BDDP composed of divergently-arranged core promoters separated by duplicated enhancers, all derived from a single promoter sequence, can be used to significantly enhance transgene expression and to direct synchronized expression of multiple transgenes. Publication Types: Research Support, Non-U.S. Gov't PMID: 15198202 [PubMed - indexed for MEDLINE] 1247: Transgenic Res. 2004 Apr;13(2):109-18. A ROS repressor-mediated binary regulation system for control of gene expression in transgenic plants. Schäfer UA, Hegedus DD, Bate NJ, Hannoufa A. Molecular Genetics Section, Agriculture and Agri-Food Canada, Saskatoon Research Centre, 107 Science Place, Saskatoon, Sask., Canada S7N 0X2. We describe a novel binary system to control transgene expression in plants. The system is based on the prokaryotic repressor, ROS, from Agrobacterium tumefaciens, optimized for plant codon usage and for nuclear targeting (synROS). The ROS protein bound in vitro to double stranded DNA comprising the ROS operator sequence, as well as to single stranded ROS operator DNA sequences, in an orientation-independent manner. A synROS-GUS fusion protein was localized to the nucleus, whereas wtROS-GUS fusion remained in the cytoplasm. The ability of synROS to repress transgene expression was validated in transgenic Arabidopsis thaliana and Brassica napus. When expressed constitutively under the actin2 promoter, synROS repressed the expression of the reporter gene gusA linked to a modified CaMV35S promoter containing ROS operator sequences in the vicinity of the TATA box and downstream of the transcription initiation signal. Repression ranged from 32 to 87% in A. thaliana, and from 23 to 76% in B. napus. These results are discussed in relation to the potential application of synROS in controlling the expression of transgenes and endogenous genes in plants and other organisms. PMID: 15198199 [PubMed - indexed for MEDLINE] 1248: Proc Natl Acad Sci U S A. 2004 Jun 22;101(25):9303-8. Epub 2004 Jun 10. Population effects of growth hormone transgenic coho salmon depend on food availability and genotype by environment interactions. Devlin RH, D'Andrade M, Uh M, Biagi CA. Fisheries and Oceans Canada, 4160 Marine Drive, West Vancouver, BC, Canada V7V 1N6. devlinr@dfo-mpo.gc.ca Environmental risk assessment of genetically modified organisms requires determination of their fitness and invasiveness relative to conspecifics and other ecosystem members. Cultured growth hormone transgenic coho salmon (Oncorhynchus kisutch) have enhanced feeding capacity and growth, which can result in large enhancements in body size (>7-fold) relative to nontransgenic salmon, but in nature, the ability to compete for available food is a key factor determining survival fitness and invasiveness of a genotype. When transgenic and nontransgenic salmon were cohabitated and competed for different levels of food, transgenic salmon consistently outgrew nontransgenic fish and could affect the growth of nontransgenic cohorts except when food availability was high. When food abundance was low, dominant individuals emerged, invariably transgenic, that directed strong agonistic and cannibalistic behavior to cohorts and dominated the acquisition of limited food resources. When food availability was low, all groups containing transgenic salmon experienced population crashes or complete extinctions, whereas groups containing only nontransgenic salmon had good (72.0 +/- 4.3% SE) survival, and their population biomass continued to increase. Thus, effects of growth hormone transgenic salmon on experimental populations were primarily mediated by an interaction between food availability and population structure. These data, while indicative of forces which may act on natural populations, also underscore the importance of genotype by environment interactions in influencing risk assessment data for genetically modified organisms and suggest that, for species such as salmon which are derived from large complex ecosystems, considerable caution is warranted in applying data from individual studies. Publication Types: Comparative Study Research Support, Non-U.S. Gov't PMID: 15192145 [PubMed - indexed for MEDLINE] 1249: EMBO Rep. 2004 May;5(5):432-6. GM food and crops: what went wrong in the UK? Many of the public's concerns have little to do with science. Burke D. UK Advisory Committee on Novel Foods and Processes. dcb27@cam.ac.uk PMID: 15184970 [PubMed - indexed for MEDLINE] 1250: Appl Environ Microbiol. 2004 Jun;70(6):3239-45. Metabolism of zearalenone by genetically modified organisms expressing the detoxification gene from Clonostachys rosea. Takahashi-Ando N, Ohsato S, Shibata T, Hamamoto H, Yamaguchi I, Kimura M. Laboratory for Remediation Research, Plant Science Center, RIKEN, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan. Zearalenone (ZEN) is converted to a nontoxic product by a lactonohydololase encoded by zhd101. An enhanced green fluorescent protein (EGFP) gene was fused to zhd101 (i.e., egfp::zhd101) and expressed in Escherichia coli. Both recombinant ZHD101 and EGFP::ZHD101 were purified to homogeneity and characterized. Maximal activity of ZHD101 toward ZEN was measured at approximately 37 to 45 degrees C and pH 10.5 (k(cat) at 30 degrees C, 0.51 s(-1)). The enzyme was irreversibly inactivated at pH values below 4.5 or by treatment with serine protease inhibitors. ZHD101 was also active against five ZEN cognates, although the efficiencies were generally low; e.g., the k(cat) was highest with zearalanone (1.5 s(-1)) and lowest with beta-zearalenol (0.075 s(-1)). EGFP::ZHD101 had properties similar to those of the individual proteins with regard to the EGFP fluorescence and lactonohydrolase activity. Fortuitously, EGFP::ZHD101 exhibited a good correlation between the fluorescence intensity and reaction velocity under various pH conditions. We therefore used egfp::zhd101 to visually monitor the lactonohydrolase activity in genetically modified organisms and evaluated the usefulness of zhd101 for in vivo detoxification of ZEN. While recombinant E. coli and transgenic rice calluses exhibited strong EGFP fluorescence and completely degraded ZEN in liquid media, recombinant Saccharomyces cerevisiae gave poor fluorescence and did not eliminate all the toxicity of the mycotoxin in the medium; i.e., the rest of ZEN was transformed into an unfavorable substrate, beta-zearalenol, by an as-yet-unidentified reductase and remained in the medium. Even so, as much as 75% of ZEN was detoxified by the yeast transformant, which is better than the detoxification system in which food-grade Lactobacillus strains are used (H. El-Nezami, N. Polychronaki, S. Salminen, and H. Mykkuäne, Appl. Environ. Microbiol. 68:3545-3549, 2002). An appropriate combination of a candidate host microbe and the codon-optimized synthetic gene may contribute significantly to establishing a mycotoxin detoxification system for food and feed. Publication Types: Evaluation Studies Research Support, Non-U.S. Gov't PMID: 15184117 [PubMed - indexed for MEDLINE] 1251: Anal Biochem. 2004 Jul 1;330(1):52-7. A cell-free biosensor for the detection of transcriptional inducers using firefly luciferase as a reporter. Pellinen T, Huovinen T, Karp M. Department of Biochemistry and Food chemistry, University of Turku, Tykistökatu 6, FIN-20520 Turku, Finland. A cell-free biosensor for the detection of transcription induction by specific small-molecule ligands is presented. As model systems, tetracycline and mercury-inducible promoters were used containing firefly luciferase as reporter gene. Escherichia coli S30 extract was prepared and used for coupled transcription-translation reactions. By using purified Tet repressor and MerR regulatory proteins, we could study repressor-operator interactions for optimizing the relative concentrations of each component. Previously, detection of tetracycline and mercury using similar transcriptional regulation in whole living cells has been carried out. As compared to whole-cell biosensors, our results showed better sensitivity for the detection of tetracycline and the toxic effect of mercury was avoided in the cell-free system. Also, as the system omits cell cultivation and bacterial membranes as molecule passage inhibitors, it is possible to carry out assays in much shorter times and without the use of genetically modified organisms. Publication Types: Research Support, Non-U.S. Gov't PMID: 15183761 [PubMed - indexed for MEDLINE] 1252: J Exp Zoolog A Comp Exp Biol. 2004 Jun 1;301(6):477-90. Growth enhancement and food conversion efficiency of transgenic fish Labeo rohita. Venugopal T, Anathy V, Kirankumar S, Pandian TJ. Department of Genetics, Center for Advanced Studies in Functional Genomics, School of Biological Sciences, Madurai Kamaraj University, Madurai 625 021 India. Three family lines of fast growing transgenic rohu Labeo rohita (rohu) were generated by electroporated-sperm-mediated transfer of the vectors harboring CMV promoter or grass carp beta-actin promoter fused to endogenous rohu GH (rGH) cDNA. The gene transfer efficiency was 25%. The transgenic rohu (family line 1) with CMV promoter showed a growth enhancement of four times normal size, whereas those (family lines 2 and 3) generated with beta-actin promoter grew 4.5 and 5.8 times faster than their respective control siblings. Southern analysis confirmed the transgene extrachromosomal (Te) persistence until the 60th week in family 1. The individuals of family lines 2 and 3, however, showed integration (Ti), as well as persistence as extrachromosomal copies (Te) until the age of 30 weeks. Mosaicism of the transgene was shown at the levels of its presence and expression. The ectopic expression of rGH mRNA was confirmed by RT-PCR. Feeding experiments revealed that the transgenic rohu ate food at a lower rate but grew more efficiently than their control siblings. Copyright 2004 Wiley-Liss, Inc. Publication Types: Comparative Study Research Support, Non-U.S. Gov't PMID: 15181642 [PubMed - indexed for MEDLINE] 1253: Mini Rev Med Chem. 2004 Jun;4(5):493-503. The genetics and properties of cereal ribosome-inactivating proteins. Motto M, Lupotto E. Istituto sperimentale per la Cerealicoltura, via Stezzano 24, 24126 Bergamo, Italy. motto@iscbg.it Plants contain proteins that are capable of inactivating ribosomes, commonly referred to as Ribosome Inactivating Proteins (RIPs). These particular plant proteins have received attention in biological and biomedical research because of their unique biological activities towards animals and human cells as cell-killing agents. Some of the best-characterised RIPs have been isolated from exotic plants, but they have also been found in cereals and other food crops. Cereals contain, in general, RIPs in the endosperm protein pool: they share a high similarity with all the other RIPs retaining, however, characteristic features forming a distinct class which diversified significantly during evolution. They appear to be involved in quite different physiological roles, such as defence against pathogens and/or involved in regulatory and developmental processes. This review aims to provide a critical assessment to work related to cereal RIP with particular emphasis to the maize RIPs. Publication Types: Review PMID: 15180506 [PubMed - indexed for MEDLINE] 1254: Nature. 2004 Jun 3;429(6991 Suppl):10-3. Farmers to pharmas. Collis B. PMID: 15175705 [PubMed - indexed for MEDLINE] 1255: Nat Biotechnol. 2004 Jun;22(6):680-2. Comment on: Nat Biotechnol. 2004 Jun;22(6):739-45. From alpha to omega-producing essential fatty acids in plants. Green AG. Food Futures, CSIRO Plant Industry, Canberra, ACT, Black Mountain, ACT, Australia. allan.green@csiro.au Publication Types: Comment News PMID: 15175687 [PubMed - indexed for MEDLINE] 1256: Nat Biotechnol. 2004 Jun;22(6):654-5. Comment on: Nat Biotechnol. 2004 Feb;22(2):204-9. Chasing 'transgenic' shadows. Miller HI, Conko G. Publication Types: Comment Letter PMID: 15175675 [PubMed - indexed for MEDLINE] 1257: Nat Biotechnol. 2004 Jun;22(6):642. China ramps up efforts to commercialize GM rice. Jia H, Jayaraman KS, Louët S. Publication Types: News PMID: 15175669 [PubMed - indexed for MEDLINE] 1258: Nat Biotechnol. 2004 Jun;22(6):637. Playing catch-up. [No authors listed] Publication Types: Editorial PMID: 15175666 [PubMed - indexed for MEDLINE] 1259: Planta. 2004 Sep;219(5):790-6. Epub 2004 Jun 2. Glyphosate inhibits the translocation of green fluorescent protein and sucrose from a transgenic tobacco host to Cuscuta campestris Yunk. Nadler-Hassar T, Goldshmidt A, Rubin B, Wolf S. R.H. Smith Institute of Plant Sciences and Genetics in Agriculture, Faculty of Agricultural, Food and Environmental Quality Sciences, The Hebrew University of Jerusalem, 76100, Rehovot, Israel. The parasitic plant Cuscuta campestris is dependent on its host for water, assimilates and amino acids. It can be controlled by the herbicide glyphosate, which inhibits 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS), resulting in shikimate accumulation. In this study, C. campestris was parasitic on transgenic tobacco plants expressing green fluorescent protein (GFP) in the phloem. Changes in [14C]sucrose and GFP accumulation in the parasite were used as indicators of the herbicide's effect on translocation between the host and parasite. Host plants were treated with glyphosate 22 days after sowing. Shikimate accumulation in the parasite 1 day after glyphosate treatment (DAGT) confirmed EPSPS inhibition in C. campestris. No damage was visible in the host plants for the first 3 DAGT, while during that same time, a significant reduction in [14C]sucrose and GFP accumulation was observed in the parasite. Thus, we propose that the parallel reduction in GFP and sucrose accumulation in C. campestris is a result of a glyphosate effect on the parasite's ability to withdraw assimilates from the host. Publication Types: Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. PMID: 15173945 [PubMed - indexed for MEDLINE] 1260: Int J Obes Relat Metab Disord. 2004 Jul;28(7):858-69. Role of parents in the determination of the food preferences of children and the development of obesity. Benton D. Department of Psychology, University of Wales Swansea, Swansea, Wales, UK. d.benton@swansea.ac.uk The role of parental behaviour in the development of food preferences is considered. Food preferences develop from genetically determined predispositions to like sweet and salty flavours and to dislike bitter and sour tastes. Particularly towards the second year of life, there is a tendency to avoid novel foods (neophobia). Food aversions can be learnt in one trial if consumption is followed by discomfort. There is a predisposition to learn to like foods with high-energy density. However, from birth genetic predispositions are modified by experience and in this context during the early years parents play a particularly important role. Parental style is a critical factor in the development of food preferences. Children are more likely to eat in emotionally positive atmospheres. Siblings, peers and parents can act as role models to encourage the tasting of novel foods. Repeated exposure to initially disliked foods can breakdown resistance. The offering of low-energy-dense foods allows the child to balance energy intake. Restricting access to particular foods increases rather than decreases preference. Forcing a child to eat a food will decrease the liking for that food. Traditionally, educational strategies have typically involved attempts to impart basic nutritional information. Given the limited ability of information to induce changes in behaviour, an alternative strategy would be to teach parents about child development in the hope that an understanding of the characteristic innate tendencies and developmental stages can be used to teach healthy food preferences. Publication Types: Review PMID: 15170463 [PubMed - indexed for MEDLINE] 1261: Shokuhin Eiseigaku Zasshi. 2004 Feb;45(1):19-24. A histochemical method using a substrate of beta-glucuronidase for detection of genetically modified papaya. Wakui C, Akiyama H, Watanabe T, Fitch MM, Uchikawa S, Ki M, Takahashi K, Chiba R, Fujii A, Hino A, Maitani T. National Institute of Health Sciences: 1-18-1, Kamiyoga, Setagaya-ku, Tokyo 158-8501, Japan. A histochemical assay for detecting genetically modified (GM) papaya (derived from Line 55-1) is described. GM papaya, currently undergoing a safety assessment in Japan, was developed using a construct that included a beta-glucuronidase (GUS) reporter gene linked to a virus coat protein (CP) gene. Histochemical assay was used to visualize the blue GUS reaction product from transgenic seed embryos. Twelve embryos per fruit were extracted from the papaya seeds using a surgical knife. The embryos were incubated with the substrate 5-bromo-4-chloro-3-indolyl-beta-D-glucuronide (X-Gluc) in a 96-well microtiter plate for 10-15 hours at 37 degrees C. Seventy-five percent of GM papaya embryos should turn blue theoretically. The histochemical assay results were completely consistent with those from a qualitative polymerase chain reaction (PCR) method developed by this laboratory. Furthermore, the method was validated in a five-laboratory study. The method for detection of GM papaya is rapid and simple, and does not require use of specialized equipment. Publication Types: Research Support, Non-U.S. Gov't PMID: 15168556 [PubMed - indexed for MEDLINE] 1262: Nature. 2004 May 27;429(6990):330. Monsanto wins seven-year court battle for seed patent. Spurgeon D. Publication Types: News PMID: 15164024 [PubMed - indexed for MEDLINE] 1263: J Agric Food Chem. 2004 Jun 2;52(11):3275-80. Erratum in: J Agric Food Chem. 2004 Jun 30;52(13):4350. Development of a seven-target multiplex PCR for the simultaneous detection of transgenic soybean and maize in feeds and foods. Germini A, Zanetti A, Salati C, Rossi S, Forré C, Schmid S, Marchelli R, Fogher C. Dipartimento di Chimica Organica e Industriale, Università di Parma, Italy. The detection of genetically modified organisms (GMOs) in food and feed is an important issue for all the subjects involved in raw material control, food industry, and distribution. Because the number of GMOs authorized in the EU increased during the past few years, there is a need for methods that allow a rapid screening of products. In this paper, we propose a method for the simultaneous detection of four transgenic maize (MON810, Bt11, Bt 176, and GA21) and one transgenic soybean (Roundup Ready), which allows routine control analyses to be sped up. DNA was extracted either from maize and soybean seeds and leaves or reference materials, and the recombinant DNA target sequences were detected with 7 primer pairs, accurately designed to be highly specific for each investigated transgene. Cross and negative controls were performed to ensure the specificity of each primer pair. The method was validated on an interlaboratory ring test and good analytical parameters were obtained (LOD = 0.25%, Repeatability, (r) = 1; Reproducibility, (R) = 0.9). The method was then applied to a model biscuit made of transgenic materials baked for the purpose and to real samples such as feed and foodstuffs. On account of the high recognition specificity and the good detection limits, this multiplex PCR represents a fast and reliable screening method directly applicable in all the laboratories involved in raw material and food control. Publication Types: Research Support, Non-U.S. Gov't PMID: 15161182 [PubMed - indexed for MEDLINE] 1264: Trends Biotechnol. 2004 Jun;22(6):286-94. The contribution of farm animals to human health. Kues WA, Niemann H. Department of Biotechnology, Institut für Tierzucht, Mariensee, D-31535 Neustadt, Germany. Farm animals and their products have a longstanding and successful history of providing significant contributions to human nutrition, clothing, facilitation of labour, research, development and medicine and have thus been essential in improving life expectancy and human health. With the advent of transgenic technologies the potential of farm animals for improving human health is growing and many areas remain to be explored. Recent breakthroughs in reproductive technologies, such as somatic cloning and in vitro embryo production, and their merger with molecular genetic tools, will further advance progress in this field. Here, we have summarized the contribution of farm animals to human health, covering the production of antimicrobial peptides, dietary supplements or functional foods, animals used as disease models and the contribution of animals to solving urgent environmental problems and challenges in medicine such as the shortage of human cells, tissues and organs and therapeutic proteins. Some of these areas have already reached the level of preclinical testing or commercial application, others will be further advanced only when the genomes of the animals concerned have been sequenced and annotated. Provided the necessary precautions are being taken, the transmission of pathogens from animals to humans can be avoided to provide adequate security. Overall, the promising perspectives of farm animals and their products warrant further research and development in this field. Publication Types: Review PMID: 15158058 [PubMed - indexed for MEDLINE] 1265: Acta Chir Belg. 2004 Apr;104(2):132-9. Evolutionary medicine. Swynghedauw B. U542-INSERM, Hôpital Lariboisière, Paris, France. Bernard.Swynghedauw@larib.inserm.fr Nothing in biology makes sense except in the light of evolution. Evolutionary, or darwinian, medicine takes the view that contemporary diseases result from incompatibility between the conditions under which the evolutionary pressure had modified our genetic endowment and the lifestyle and dietary habits in which we are currently living, including the enhanced lifespan, the changes in dietary habits and the lack of physical activity. An evolutionary trait express a genetic polymorphism which finally improve fitness, it needs million years to become functional. A limited genetic diversity is a necessary prerequisite for evolutionary medicine. Nevertheless, search for a genetic endowment would become nearly impossible if the human races were genetically different. From a genetic point of view, homo sapiens, is homogeneous, and the so-called human races have only a socio-economic definition. Historically, Heart Failure, HF, had an infectious origin and resulted from mechanical overload which triggered mechanoconversion by using phylogenically ancient pleiotropic pathways. Adaptation was mainly caused by negative inotropism. Recently, HF was caused by a complex remodelling caused by the trophic effects of mechanics, ischemia, senescence, diabetes and, neurohormones. The generally admitted hypothesis is that cancers were largely caused by a combination of modern reproductive and dietary lifestyles mismatched with genotypic traits, plus the longer time available for a confrontation. Such a concept is illustrated for skin and breast cancers, and also for the link between cancer risk and dietary habits. Publication Types: Review PMID: 15154569 [PubMed - indexed for MEDLINE] 1266: Bull Entomol Res. 2004 Apr;94(2):179-88. An interspersed refuge for Sitodiplosis mosellana (Diptera: Cecidomyiidae) and a biocontrol agent Macroglenes penetrans (Hymenoptera: Pteromalidae) to manage crop resistance in wheat. Smith MA, Lamb RJ, Wise IL, Olfert OO. Cereal Research Centre Agriculture and Agri-Food Canada 195 Dafoe Road Winnipeg, Manitoba, R3T 2M9 Canada. An interspersed refuge of susceptible plants in a resistant, spring-sown wheat crop was tested as a strategy to protect crop resistance against evolution of virulence by the wheat midge Sitodiplosis mosellana (Géhin), and also to conserve a biocontrol agent Macroglenes penetrans(Kirby). Eight replicated field experiments were conducted using seed mixtures of 0, 5, 10, 15 and 100% or 0, 5 and 100% susceptible wheat with an agronomically similar wheat expressing the antibiotic resistance gene Sm1. The frequencies of eggs, mature larvae and parasitized larvae in susceptible and resistant wheat spikes, and midge-affected seeds in the harvest, were recorded for each plot. In susceptible wheat, insect densities and seed damage were typical of those in commercial wheat. In resistant wheat, few larvae completed development, 2% or less compared with about 80% in susceptible wheat, when larvae were sampled at maturity. This resistant wheat also deterred midge oviposition, reducing egg densities by 65% compared with susceptible wheat. The wheat midge and its parasitoid oviposited throughout the plots, and parasitism was density independent. The densities of mature midge larvae and parasitoids were in proportion to the size of the refuge. A 5% susceptible refuge produced about 41 mature larvae for each mature larva from the resistant wheat, and provided effective control of damage. An interspersed refuge of susceptible plants in resistant wheat is a promising strategy for sustaining resistance conferred by Sm1 and biocontrol of the wheat midge. Publication Types: Research Support, Non-U.S. Gov't PMID: 15153300 [PubMed - indexed for MEDLINE] 1267: J Allergy Clin Immunol. 2004 May;113(5):1003-4; author reply 1004-5. Comment in: J Allergy Clin Immunol. 2004 Jul;114(1):127-30. Comment on: J Allergy Clin Immunol. 2003 Nov;112(5):1011-2. Starlink genetically modified corn and allergenicity in an individual. Siruguri V, Sesikeran B, Bhat RV. Publication Types: Case Reports Comment Letter PMID: 15148966 [PubMed - indexed for MEDLINE] 1268: Nahrung. 2004 Apr;48(2):149-55. Sensory and rheological properties of transgenically and chemically modified starch ingredients as evaluated in a food product model. Ahmt T, Wischmann B, Blennow A, Madsen F, Bandsholm O, Thomsen J. Biotechnological Institute, Dept. of Perception & Functionality, Holbergsvej 10, DK-6000 Kolding, Denmark. tina.ahmt@teknologisk.dk Starches derived from five genetically modified potato lines, two chemically modified potato starches and two native starches from potato and maize were subjected to physical and chemical analyses and their functionality evaluated in a milk-based food product model. The transgenic starches were specifically modified with respect to amylopectin chain length and phosphorous content by suppression of the starch branching enzyme and overexpression of glycogen branching enzyme. Transgenic starches with long amylopectin chains and high phosphorous content had increased gelatinisation temperatures, produced gels with a higher tendency to retrograde and a low freeze/thaw stability as compared to starches with shorter amylopectin chains and lower phosphorous content. The textural properties of the food product model prepared from genetically and chemically modified starches were characterised by sensory and rheological analyses. To clearly visualise the effects of the modifications, data was evaluated by radar plots and multiple regression analysis (chemometrics). Genetically modified potato starches with longer amylopectin chains and increased phosphorous content gave a more gelled and a shorter texture as compared to starches with shorter amylopectin chains and decreased phosphorous content. Acetylated and hydroxypropylated potato starches gave sticky and stringy textures. Correlations between rheology parameters and sensory parameters were found. The sensory parameter stringy/long could be predicted from the rheological data. Publication Types: Research Support, Non-U.S. Gov't PMID: 15146974 [PubMed - indexed for MEDLINE] 1269: Clin Med. 2004 Mar-Apr;4(2):193-4. GM foods: is rational public debate possible? [No authors listed] Publication Types: Interview PMID: 15139747 [PubMed - indexed for MEDLINE] 1270: Br J Nutr. 2004 May;91(5):673-81. Relative stability of transgene DNA fragments from GM rapeseed in mixed ruminal cultures. Sharma R, Alexander TW, John SJ, Forster RJ, McAllister TA. Agriculture and Agri-Food Canada Research Center, PO Box 3000, Lethbridge, Alberta Canada T1J 4B1. The use of transgenic crops as feeds for ruminant animals has prompted study of the possible uptake of transgene fragments by ruminal micro-organisms and/or intestinal absorption of fragments surviving passage through the rumen. The persistence in buffered ruminal contents of seven different recombinant DNA fragments from GM rapeseed expressing the 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) transgene was tracked using PCR. Parental and transgenic (i.e. glyphosphate-tolerant; Roundup Ready, Monsanto Company, St Louis, MO, USA) rapeseed were incubated for 0, 2, 4, 8, 12, 24 and 48 h as whole seeds, cracked seeds, rapeseed meal, and as pelleted, barley-based diets containing 65 g rapeseed meal/kg. The seven transgene fragments ranged from 179 to 527 bp and spanned the entire 1363 bp EPSPS transgene. A 180 bp ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) small subunit fragment and a 466 bp 16S rDNA fragment were used as controls for endogenous rapeseed DNA and bacterial DNA respectively. The limit of detection of the PCR assay, established using negative controls spiked with known quantities of DNA, was 12.5 pg. Production of gas and NH3 was monitored throughout the incubation and confirmed active in vitro fermentation. Bacterial DNA was detected in all sample types at all time points. Persistence patterns of endogenous (Rubisco) and recombinant (EPSPS) rapeseed DNA were inversely related to substrate digestibility (amplifiable for 48, 8 and 4 h in whole or cracked seeds, meal and diets respectively), but did not differ between parental and GM rapeseed, nor among fragments. Detection of fragments was representative of persistence of the whole transgene. No EPSPS fragments were amplifiable in microbial DNA, suggesting that transformation had not occurred during the 48 h incubation. Uptake of transgenic DNA fragments by ruminal bacteria is probably precluded or time-limited by rapid degradation of plant DNA upon plant cell lysis. Publication Types: Research Support, Non-U.S. Gov't PMID: 15137918 [PubMed - indexed for MEDLINE] 1271: Eur Ann Allergy Clin Immunol. 2004 Mar;36(3):88-91. [Animal models for assessment of GMO allergenicity: advantages and limitations] [Article in French] Adel-Patient K, Wal JM. Laboratoire d'Immuno-Allergie Alimentaire, INRA, CEA-Service de Pharmacologie et d'Immunologie CEA de Saclay, 91191 Gif-sur-Yvette. Incidence of IgE-mediated allergic reactions to foods is increasing as well as the severity of associated symptoms and numerous foods are now incriminated, probably in relation with modifications of dietary habits and increased exposure to new or modified food ingredients. Therefore, the introduction on the market of food composed of or derived from genetically modified organisms (GMOs) raised the question of their potential allergenicity. Particularly with regards to the allergenicity of a newly expressed protein, it is necessary to obtain, from several steps in the risk assessment process, a cumulative body of evidence which minimises any uncertainty. This may include the use of animal model despite no fully reliable validated model is available yet. Such animal models should allow to address 3 major issues: Is the novel protein a sensitizer, i.e. does it possess intrinsic properties that allow to sensitize a predisposed individual? Is the protein an elicitor i.e. is it able to elicit an allergic reaction in a sensitised individual? And is the protein an adjuvant, i.e. can it facilitate or enhance the sensitisation to an other protein? Animal models under investigation currently include mice, rats and guinea pigs but models such as dogs and swine also appeared a few years ago. The aim is to mimic the mechanism and characteristics of the sensitisation phase and/or the elicitation phase of the allergic reaction as it occurs in atopic humans. They are necessary because sensitisation studies can obviously not be done in human and because in vitro tests cannot reproduce the complexity of the immune system. We propose a mouse model which mimics both phases of the allergic reaction. It has permitted to evidence that biochemical and clinical manifestations occuring during the active phases of the allergic reaction differ according to the structure of the allergen used for the challenge. This may allow to compare the allergenic potential of a genetically modified protein with that of the conventional one and to identify possible unintended effects. However, pathogenesis of food allergy in human is very complex and multifactorial, including individual differences in susceptibility, environmental factors, conditions of exposure, ... No animal model can take into account all these factors and allow a reliable prediction of the prevalence and severity of allergic reactions which would result from the exposure to a (novel) protein. Nevertheless, point by point analysis using the different models available may provide useful informations on the potential allergenicity of a novel protein. Publication Types: English Abstract Review PMID: 15137477 [PubMed - indexed for MEDLINE] 1272: J Biotechnol. 2004 Mar 4;108(2):179-83. Subcellular targeting of human interleukin-10 in plants. Menassa R, Kennette W, Nguyen V, Rymerson R, Jevnikar A, Brandle J. Southern Crop Protection and Food Research Centre, Agriculture and Agri-Food Canada, 1391 Sandford Street, London, Ont., Canada N5V 4T3. The utility of plants for the production of a wide range of recombinant proteins is now clearly established. However, the challenge remains to produce these proteins at sufficient concentrations for extraction to be economically feasible. In this paper, we have investigated the ability of plant cells to accumulate the human interleukin-10 (IL-10) protein targeted to chloroplasts and mitochondria. We found that IL-10 accumulates in chloroplasts only if a 6 x His tag is added at the C-terminus of the protein. The hexapeptide may provide protection from degradation. Conversely, the IL-10 protein does not accumulate in mitochondria. Analysis of the chloroplast-targeted IL-10 protein revealed only monomeric IL-10 and limited biological activity in in vitro cell assays. Publication Types: Comparative Study Research Support, Non-U.S. Gov't PMID: 15129727 [PubMed - indexed for MEDLINE] 1273: Food Chem Toxicol. 2004 Jul;42(7):1195-202. Safety assessment, detection and traceability, and societal aspects of genetically modified foods. European Network on Safety Assessment of Genetically Modified Food Crops (ENTRANSFOOD). Concluding remarks. Kuiper HA, König A, Kleter GA, Hammes WP, Knudsen I; European Network on Safety Assessment of Genetically Modified Food Crops (ENTRANSFOOD). RIKILT-Institute of Food Safety, Wageningen University & Research Centre, Bornsesteeg 45, PO Box 230, NL-6700 AE Wageningen, The Netherlands. harry.kuiper@wur.nl The most important results from the EU-sponsored ENTRANSFOOD Thematic Network project are reviewed, including the design of a detailed step-wise procedure for the risk assessment of foods derived from genetically modified crops based on the latest scientific developments, evaluation of topical risk assessment issues, and the formulation of proposals for improved risk management and public involvement in the risk analysis process. Copyright 2004 Elsevier Ltd. Publication Types: Review PMID: 15123387 [PubMed - indexed for MEDLINE] 1274: Food Chem Toxicol. 2004 Jul;42(7):1181-93. Societal aspects of genetically modified foods. Frewer L, Lassen J, Kettlitz B, Scholderer J, Beekman V, Berdal KG. Marketing and Consumer Behaviour Group, University of Wageningen, Wageningen University & Research Centre, Hollandseweg 1, 6706 KN Wageningen, The Netherlands. lynn.frewer@wur.nl This paper aims to examine some of the reasons behind public controversy associated with the introduction of genetically modified foods in Europe the 1990s. The historical background to the controversy is provided to give context. The issue of public acceptance of genetically modified foods, and indeed the emerging biosciences more generally, is considered in the context of risk perceptions and attitudes, public trust in regulatory institutions, scientists, and industry, and the need to develop communication strategies that explicitly include public concerns rather than exclude them. Increased public participation has been promoted as a way of increasing trust in institutional practices associated with the biosciences, although questions still arise as to how to best utilise the outputs of such exercises in policy development. This issue will become more of a priority as decision-making systems become more transparent and open to public scrutiny. The results are discussed in the context of risk assessment and risk management, and recommendations for future research are made. In particular, it is recommended that new methods are developed in order to integrate public values more efficaciously into risk analysis processes, specifically with respect to the biosciences and to technology implementation in general. Copryright 2004 Elsevier Ltd. Publication Types: Multicenter Study Review PMID: 15123386 [PubMed - indexed for MEDLINE] 1275: Food Chem Toxicol. 2004 Jul;42(7):1157-80. Detection and traceability of genetically modified organisms in the food production chain. Miraglia M, Berdal KG, Brera C, Corbisier P, Holst-Jensen A, Kok EJ, Marvin HJ, Schimmel H, Rentsch J, van Rie JP, Zagon J. Istituto Superiore di Sanitá, Section of Cereal Chemistry, Laboratory of Food, Viale Regina Elena 299, I-00161 Rome, Italy. Both labelling and traceability of genetically modified organisms are current issues that are considered in trade and regulation. Currently, labelling of genetically modified foods containing detectable transgenic material is required by EU legislation. A proposed package of legislation would extend this labelling to foods without any traces of transgenics. These new legislations would also impose labelling and a traceability system based on documentation throughout the food and feed manufacture system. The regulatory issues of risk analysis and labelling are currently harmonised by Codex Alimentarius. The implementation and maintenance of the regulations necessitates sampling protocols and analytical methodologies that allow for accurate determination of the content of genetically modified organisms within a food and feed sample. Current methodologies for the analysis of genetically modified organisms are focused on either one of two targets, the transgenic DNA inserted- or the novel protein(s) expressed- in a genetically modified product. For most DNA-based detection methods, the polymerase chain reaction is employed. Items that need consideration in the use of DNA-based detection methods include the specificity, sensitivity, matrix effects, internal reference DNA, availability of external reference materials, hemizygosity versus homozygosity, extrachromosomal DNA, and international harmonisation. For most protein-based methods, enzyme-linked immunosorbent assays with antibodies binding the novel protein are employed. Consideration should be given to the selection of the antigen bound by the antibody, accuracy, validation, and matrix effects. Currently, validation of detection methods for analysis of genetically modified organisms is taking place. In addition, new methodologies are developed, including the use of microarrays, mass spectrometry, and surface plasmon resonance. Challenges for GMO detection include the detection of transgenic material in materials with varying chromosome numbers. The existing and proposed regulatory EU requirements for traceability of genetically modified products fit within a broader tendency towards traceability of foods in general and, commercially, towards products that can be distinguished from each other. Traceability systems document the history of a product and may serve the purpose of both marketing and health protection. In this framework, segregation and identity preservation systems allow for the separation of genetically modified and non-modified products from "farm to fork". Implementation of these systems comes with specific technical requirements for each particular step of the food processing chain. In addition, the feasibility of traceability systems depends on a number of factors, including unique identifiers for each genetically modified product, detection methods, permissible levels of contamination, and financial costs. In conclusion, progress has been achieved in the field of sampling, detection, and traceability of genetically modified products, while some issues remain to be solved. For success, much will depend on the threshold level for adventitious contamination set by legislation. Copryright 2004 Elsevier Ltd. Publication Types: Multicenter Study Research Support, Non-U.S. Gov't Review PMID: 15123385 [PubMed - indexed for MEDLINE] 1276: Food Chem Toxicol. 2004 Jul;42(7):1127-56. The relevance of gene transfer to the safety of food and feed derived from genetically modified (GM) plants. van den Eede G, Aarts H, Buhk HJ, Corthier G, Flint HJ, Hammes W, Jacobsen B, Midtvedt T, van der Vossen J, von Wright A, Wackernagel W, Wilcks A. European Commission Directorate General Joint Research Centre, Institute for Health and Consumer Protection, Biotechnology and GMOs Unit, Via E. Fermi 1-T.P. 331, I-21020 Ispra (VA), Italy. guy.van-den-eede@cec.eu.int In 2000, the thematic network ENTRANSFOOD was launched to assess four different topics that are all related to the testing or assessment of food containing or produced from genetically modified organisms (GMOs). Each of the topics was linked to a European Commission (EC)-funded large shared cost action (see http://www.entransfood.com). Since the exchange of genetic information through horizontal (lateral) gene transfer (HGT) might play a more important role, in quantity and quality, than hitherto imagined, a working group dealing with HGT in the context of food and feed safety was established. This working group was linked to the GMOBILITY project (GMOBILITY, 2003) and the results of the deliberations are laid down in this review paper. HGT is reviewed in relation to the potential risks of consuming food or feed derived from transgenic crops. First, the mechanisms for obtaining transgenic crops are described. Next, HGT mechanisms and its possible evolutionary role are described. The use of marker genes is presented in detail as a special case for genes that may pose a risk. Furthermore, the exposure to GMOs and in particular to genetically modified (GM) deoxyribonucleic acid (DNA) is discussed as part of the total risk assessment. The review finishes off with a number of conclusions related to GM food and feed safety. The aim of this paper is to provide a comprehensive overview to assist risk assessors as well as regulators and the general public in understanding the safety issues related to these mechanisms. Copryright 2004 Elsevier Ltd. Publication Types: Multicenter Study Review PMID: 15123384 [PubMed - indexed for MEDLINE] 1277: Food Chem Toxicol. 2004 Jul;42(7):1089-125. Unintended effects and their detection in genetically modified crops. Cellini F, Chesson A, Colquhoun I, Constable A, Davies HV, Engel KH, Gatehouse AM, Kärenlampi S, Kok EJ, Leguay JJ, Lehesranta S, Noteborn HP, Pedersen J, Smith M. Metapontum Agrobios, SS Jonica Km 448.2, I-75010 Metaponto Matera, Italy. The commercialisation of GM crops in Europe is practically non-existent at the present time. The European Commission has instigated changes to the regulatory process to address the concerns of consumers and member states and to pave the way for removing the current moratorium. With regard to the safety of GM crops and products, the current risk assessment process pays particular attention to potential adverse effects on human and animal health and the environment. This document deals with the concept of unintended effects in GM crops and products, i.e. effects that go beyond that of the original modification and that might impact primarily on health. The document first deals with the potential for unintended effects caused by the processes of transgene insertion (DNA rearrangements) and makes comparisons with genetic recombination events and DNA rearrangements in traditional breeding. The document then focuses on the potential value of evolving "profiling" or "omics" technologies as non-targeted, unbiased approaches, to detect unintended effects. These technologies include metabolomics (parallel analysis of a range of primary and secondary metabolites), proteomics (analysis of polypeptide complement) and transcriptomics (parallel analysis of gene expression). The technologies are described, together with their current limitations. Importantly, the significance of unintended effects on consumer health are discussed and conclusions and recommendations presented on the various approaches outlined. Copryright 2004 Elsevier Ltd. Publication Types: Multicenter Study Review PMID: 15123383 [PubMed - indexed for MEDLINE] 1278: Food Chem Toxicol. 2004 Jul;42(7):1047-88. Assessment of the safety of foods derived from genetically modified (GM) crops. König A, Cockburn A, Crevel RW, Debruyne E, Grafstroem R, Hammerling U, Kimber I, Knudsen I, Kuiper HA, Peijnenburg AA, Penninks AH, Poulsen M, Schauzu M, Wal JM. Harvard Center for Risk Analysis, Harvard School of Public Health, Harvard University, 718 Huntington Avenue, Boston, MA 02115, USA. ariane_koenig@harvard.edu This paper provides guidance on how to assess the safety of foods derived from genetically modified crops (GM crops); it summarises conclusions and recommendations of Working Group 1 of the ENTRANSFOOD project. The paper provides an approach for adapting the test strategy to the characteristics of the modified crop and the introduced trait, and assessing potential unintended effects from the genetic modification. The proposed approach to safety assessment starts with the comparison of the new GM crop with a traditional counterpart that is generally accepted as safe based on a history of human food use (the concept of substantial equivalence). This case-focused approach ensures that foods derived from GM crops that have passed this extensive test-regime are as safe and nutritious as currently consumed plant-derived foods. The approach is suitable for current and future GM crops with more complex modifications. First, the paper reviews test methods developed for the risk assessment of chemicals, including food additives and pesticides, discussing which of these methods are suitable for the assessment of recombinant proteins and whole foods. Second, the paper presents a systematic approach to combine test methods for the safety assessment of foods derived from a specific GM crop. Third, the paper provides an overview on developments in this area that may prove of use in the safety assessment of GM crops, and recommendations for research priorities. It is concluded that the combination of existing test methods provides a sound test-regime to assess the safety of GM crops. Advances in our understanding of molecular biology, biochemistry, and nutrition may in future allow further improvement of test methods that will over time render the safety assessment of foods even more effective and informative. Copryright 2004 Elsevier Ltd. Publication Types: Multicenter Study Research Support, Non-U.S. Gov't Review PMID: 15123382 [PubMed - indexed for MEDLINE] 1279: J Agric Food Chem. 2004 May 5;52(9):2726-34. Compositional equivalency of Cry1F corn event TC6275 and conventional corn (Zea mays L.). Herman RA, Phillips AM, Collins RA, Tagliani LA, Claussen FA, Graham CD, Bickers BL, Harris TA, Prochaska LM. Dow AgroSciences LLC, 9330 Zionsville Road, Indianapolis, Indiana 46268, USA. raherman@dow.com Maize (Zea mays L.) plants have been transformed to express a Cry1F insecticidal crystal protein originally isolated from Bacillus thuringiensis Berliner. This protein controls lepidopteran pests of maize, including the European corn borer, Ostrinia nubilalis (Hübner). As part of the safety assessment for crops containing transgenes, a compositional analysis of the food and feed is conducted. This analysis is designed to detect unintended changes in the nutrient and antinutrient content of the raw commodities produced by the crop due to the insertion of the genes into the genomic DNA of the plant (pleotropic effects). Samples of transgenic and nontransgenic maize forage and grain were collected from six field sites located in the U.S. and Canada. Forage samples were analyzed for proximates and minerals, and grain was further analyzed for fatty acids, amino acids, vitamins, secondary metabolites, and antinutrients. Results demonstrated that maize expressing the Cry1F protein was equivalent to nontransgenic maize with respect to these important components. Comparison of the variability within the nontransgenic and transgenic hybrid, as compared to composition values reported in the literature, suggest that factors other than transgenes may contribute more substantially to the composition of crops. Publication Types: Comparative Study PMID: 15113183 [PubMed - indexed for MEDLINE] 1280: J Agric Food Chem. 2004 May 5;52(9):2709-14. Safety assessment by in vitro digestibility and allergenicity of genetically modified maize with an amaranth 11S globulin. Sinagawa-García SR, Rascón-Cruz Q, Valdez-Ortiz A, Medina-Godoy S, Escobar-Gutiérrez A, Paredes-López O. Departamento de Biotecnología y Bioquímica, Centro de Investigación y de Estudios Avanzados del IPN, Apdo. Postal 629, 36500 Irapuato, Gto., México. Prospective testing for allergenicity of proteins obtained from sources with no prior history of causing allergy has been difficult to perform. Thus, the objective of this work was to assess the food safety of genetically modified maize with an amaranth globulin protein termed amarantin. Transgenic maize lines evaluated showed, in relation to nontransgenic, 4-35% more protein and 0-44% higher contents of specific essential amino acids. Individual sequence analysis with known amino acid sequences, reported as allergens, showed that none of these IgE elicitors were identified in amarantin. Amarantin was digested within the first 15 min by Simulated Gastric Fluid treatment as observed by Western blot. Expressed amarantin did not induce important levels of specific IgE antibodies in BALB/c mice, as analyzed by ELISA. We conclude that the transgenic maize with amarantin is not an important allergenicity inducer, just as nontransgenic maize. Publication Types: Research Support, Non-U.S. Gov't PMID: 15113180 [PubMed - indexed for MEDLINE] 1281: Food Chem Toxicol. 2004 Jun;42(6):1003-14. Results of a 13 week safety assurance study with rats fed grain from glyphosate tolerant corn. Hammond B, Dudek R, Lemen J, Nemeth M. Monsanto Company, 800 N. Lindbergh, St Louis, MO 63167, USA. bruce.g.hammond@monsanto.com The current study presents the results of a 13 week feeding study in rats with grain from Roundup Ready corn which is tolerant to the herbicide glyphosate. Herbicide tolerance was accomplished through the introduction of cp4 epsps coding sequences into the corn genome for in planta production of CP4 EPSPS enzymes. Unlike related corn EPSPS enzymes, CP4 EPSPS enzymes are not inhibited by the herbicide glyphosate. Purina TestDiets formulated Roundup Ready corn grain into rodent diets at levels of 11 and 33% (w/w). The responses of rats fed diets containing Roundup Ready corn grain were compared to that of rats fed diets containing non-transgenic grain (controls). All diets were nutritionally balanced and conformed to Purina Mills, Inc. specifications for Certified LabDiet 5002. There were 400 rats in the study divided into 10 groups of 20 rats/sex/group. Overall health, body weight, food consumption, clinical pathology parameters (hematology, blood chemistry, urinalysis), organ weights, gross and microscopic appearance of tissues were comparable between groups fed diets containing Roundup Ready and control corn grain. This study complements extensive agronomic, compositional and farm animal feeding studies with Roundup Ready corn grain, confirming it is as safe and nutritious as existing commercial corn hybrids. Publication Types: Comparative Study PMID: 15110110 [PubMed - indexed for MEDLINE] 1282: Nature. 2004 Apr 22;428(6985):874-5. Benelux: fertile ground. Schiermeier Q. PMID: 15103381 [PubMed - indexed for MEDLINE] 1283: Nature. 2004 Apr 22;428(6985):788. Labelling laws for transgenic food come into effect. Nelson L. Publication Types: News PMID: 15103341 [PubMed - indexed for MEDLINE] 1284: Trends Plant Sci. 2004 Feb;9(2):70-5. Crop transformation and the challenge to increase yield potential. Sinclair TR, Purcell LC, Sneller CH. Agricultural Research Service, US Department of Agriculture, Agronomy Physiology Laboratory, University of Florida, Gainesville, FL 32611-0965, USA. trsincl@mail.ifas.ufl.edu Molecular transformation is commonly offered as a hope to overcome the apparent stagnation in crop yield potential. A basic understanding of the resource limits imposed on crops and the yield hierarchy going from gene expression to harvestable yield leads to a rather negative view that transformations of a few, or even of a complex of genes will result directly in major yield increases. Forty years of biochemical and physiological research illustrate the great difficulty in translating research at the basic level into improvements in crop yield. However, there are a few cases where physiological research has led to improved crop cultivars with increased yield. These successes are instructive in highlighting key elements required to achieve success in developing crop cultivars for increased yield. Publication Types: Review PMID: 15102372 [PubMed - indexed for MEDLINE] 1285: Wei Sheng Yan Jiu. 2004 Jan;33(1):77-80. [Immunotoxicologic assessment of transgenetic rice] [Article in Chinese] Chen X, Zhuo Q, Piao J, Yang X. Institute of Nutrition and Food safety, Chinese Center for Disease Control and Prevention, Beijing 100050, China. OBJECTIVE: To assess the immunotoxicologic aspect of transgenetic plant. METHODS: BALB/C mice were fed with food composed by transgenic rice (into which cowpea trypsin inhibitor gene was introduced) or nontransgenetic rice (which has the same gene composition as the transgenic rice except for the cowpea trypsin inhibitor gene) for 30 days. All food is made according to the composition of AIN-93G. In the end, all kinds of immunotoxicologic indexes of mice of every group were compared such as body weight, guts index, blood routine test, lymphocyte sort, serum antibody titter, plaque forming cell, delayed hypersensitivity response, macrophage function test. RESULTS: All immunotoxicologic indexes of mice fed either by transgenic rice or nontransgenetic rice have no differences to those of mice fed by normal food. CONCLUSION: Transgenic rice is substantially equivilent to nontransgenetic rice in immunotoxicologic aspect. Publication Types: English Abstract Research Support, Non-U.S. Gov't PMID: 15098485 [PubMed - indexed for MEDLINE] 1286: Wei Sheng Yan Jiu. 2004 Jan;33(1):74-7. [Study on the teratogenicity effects of genetically modified rich which expressed cowpea trypsin inhibitor on rats] [Article in Chinese] Zhuo Q, Chen X, Piao J, Han C. Institute of Nutrition and Food Safety, Chinese Centre for Diease Control and Prevention, Beijing 100050, China. OBJECTIVE: Rats were fed by transgenic rice which expressed insecticidal protein CpTI (cowpea trypsin inhibitor) to study if the transgenic rice possessed potential teratogenicity effects. METHODS: Weanling Wistar rats were randomly divided into four groups: transgenic rice group, non transgenic rice group, negative control group and positive control group. The diet of transgenic rice group contained 78.3% transgenic rice. The diet of non transgenic rice group contained 74.7% non transgenic rice which was the parent line of the transgenic one. The diet formula of the two control groups was AIN93G. The marco- and micronutrient content were equal in three diets. When the sexual maturation period of rats arrived, conventional teratogenicity test was performed. RESULTS: The body weight gain of pregnant rats and body weight, body length and tail length of fetal rats in transgenic rice group were significant higher than those in positive control group (P < 0.01). The malformation rate of fetus in transgenic rice group was significant lower than that in positive control group (P < 0.01). CONCLUSION: There were no significant differences of all indexes among transgenic rice group, non transgenic rice group and negative control group (P > 0.05). Judging from these results, the transgenic rice modified with CpTI was considered to have no maternal toxicity, embryotoxicity and teratogenicity effects. Publication Types: English Abstract Research Support, Non-U.S. Gov't PMID: 15098484 [PubMed - indexed for MEDLINE] 1287: J Mol Biol. 2004 Feb 20;336(3):579-87. The C. elegans ceh-36 gene encodes a putative homemodomain transcription factor involved in chemosensory functions of ASE and AWC neurons. Koga M, Ohshima Y. Department of Biology, Faculty of Sciences, Kyushu University Graduate School, 6-10-1 Hakozaki, Higashi-Ku, Fukuoka 812-8581, Japan. mkogascb@mbox.nc.kyushu-u.ac.jp Chemotaxis to water-soluble chemicals such as sodium ion is an important behavior of Caenorhabditis elegans for seeking food, and ASE chemosensory neurons have a major role in this behavior. We isolated mutants defective in chemotaxis to sodium acetate. We show here that among them ks86 had a mutation in the ceh-36 gene. ceh-36 :: gfp reporter constructs were expressed in ASE and AWC neurons. In a mutant of the che-1 gene, which encodes another transcription factor and is required for specification of ASE neurons, expression of the ceh-36 :: gfp reporter in ASE is lost. This indicates that the ceh-36 gene functions downstream of the che-1 gene in ASE. In the ceh-36(ks86) mutant, expression of the tax-2 gene encoding a cyclic nucleotide-gated channel was reduced in ASE and AWC. This affords an explanation for defects of the ceh-36 mutant in the chemotaxis mediated by ASE and AWC. When a ceh-36 cDNA was expressed in an adult ceh-36 mutant by a heat shock promoter, chemotaxis to sodium acetate was recovered. These results suggest that ceh-36 is required for functions, and not for development, of ASE. Publication Types: Research Support, Non-U.S. Gov't PMID: 15095973 [PubMed - indexed for MEDLINE] 1288: Curr Opin Biotechnol. 2004 Apr;15(2):162-5. The nutritional fortification of cereals. Poletti S, Gruissem W, Sautter C. Institute of Plant Sciences, Swiss Federal Institute of Technology, ETH Zurich, Universitätstrasse 2, CH-8092 Zurich, Switzerland. The low micronutrient content of cereals requires the fortification of food and biofortification of plants. Many laboratories are currently pursuing biofortification using breeding and genetic modification, but progress is challenged by technical hurdles and our understanding of physiological processes. Recent studies have largely been confined to the improvement of levels of iron, zinc, some vitamins and a variety of essential amino acids. Progress has been made in the accumulation of iron, zinc, and vitamins A and E in genetically modified plants. For future success in this area, many more studies will be required on the physiology of ion uptake and on the transport of vitamin precursors. Publication Types: Review PMID: 15081056 [PubMed - indexed for MEDLINE] 1289: Curr Opin Biotechnol. 2004 Apr;15(2):144-7. Generating high-yielding varieties by genetic manipulation of plant architecture. Sakamoto T, Matsuoka M. Field Production Science Center, The University of Tokyo, Nishi-Tokyo 188-0002, Japan. Despite a huge population increase since the 1960s, the green revolution more than doubled world grain production and averted large-scale famine. Food crop productivity will have to be further raised, however, because the world population is still increasing rapidly. Among several parameters associated with the increase in yield potential, genes that control plant height and tiller number (in cereal crops) have recently been identified. In addition, a promising strategy to generate semi-dwarf varieties has been developed. Recent advances in plant genome analyses and plant biotechnology will realize a second green revolution through the genetic engineering of food crops. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 15081053 [PubMed - indexed for MEDLINE] 1290: Anal Chem. 2004 Apr 15;76(8):2306-13. Quantitation of transgenic Bt event-176 maize using double quantitative competitive polymerase chain reaction and capillary gel electrophoresis laser-induced fluorescence. García-Cañas V, Cifuentes A, González R. Institute of Industrial Fermentations (CSIC), Juan de la Cierva 3, 28006 Madrid, Spain. In this work, a new procedure useful to quantitatively analyze genetically modified organisms (GMOs) in foods is described and applied to analyze transgenic Bt Event-176 maize. The method developed consists of coamplifications of specific DNA maize sequences with internal standards using quantitative competitive PCR (QC-PCR). The QC-PCR products are quantitatively analyzed using a capillary gel electrophoresis (CGE) with laser-induced fluorescence detection (LIF) method developed at our laboratory that utilizes a physically adsorbed coating. The CGE-LIF procedure allows the use of internal standards differing by only 10 bp from the original target fragments, to our knowledge, the smallest size difference that can be found in the bibliography for QC-PCR of GMOs. A spectrofluorometric procedure using ROX reference dye is proposed to solve calibration problems of input DNA concentration. It is demonstrated that the use of ROX drastically enhances the accuracy of the quantitative analysis by QC-PCR. Reproducibility of analysis times and corrected peak areas (measured as target/competitor PCR products ratio) for the CGE-LIF separations are determined to be better than 0.91 and 1.93% (RSD, n = 15) respectively, for three different days. It is shown that CGE-LIF provides better resolution and a signal/noise ratio improvement of approximately 700-fold compared to slab gel electrophoresis. The good possibilities in terms of quantitative analysis of GMOs provided by this new method are confirmed by determining the Bt Event-176 maize content in certified reference maize powder and food samples of known composition. This procedure opens the possibility for accurate quantitation of multiple GMOs in a single run. Publication Types: Research Support, Non-U.S. Gov't PMID: 15080742 [PubMed - indexed for MEDLINE] 1291: Health Risk Soc. 2001 Nov;3(3):245-59. New genetics, new ethics? Globalisation and its discontents. Glasner P, Rothman H. Science and Technology Policy Unit, Faculty of Economics and Social Science, University of the West of England, Coldharbour Lane, Bristol BS16 1QY, UK. Peter.Glasner@uwe.ac.uk The paper discusses the rapid and significant development of new genetic technologies (in health, food and agriculture) in the theoretical context of the globalisation debate. We show how the studies on the ethical, legal and social implications of biotechnological innovation have themselves emerged as an important factor in the technological innovation and product development process. Ethical considerations are becoming integral to attempts to understand techno-scientific developments in late modernity. However, ethical studies have so far been more focused on the medical rather than the overall commercial application of genomics. Their relevance to this wider context of the globalised commodification of new genetics needs to be explored. Of special significance is the introduction of ethical considerations into the debates on globalised risk, which we will explore in relation to increasing disparities between the global North and South. PMID: 15080126 [PubMed - indexed for MEDLINE] 1292: Risk Anal. 2004 Apr;24(2):491-9. The precautionary principle and/or risk assessment in World Trade Organization decisions: a possible role for risk perception. Goldstein B, Carruth RS. University of Pittsburgh Graduate School of Public Health, Pittsburgh, PA 15261, USA. Risk analysis has been recognized and validated in World Trade Organization (WTO) decision processes. In recent years the precautionary principle has been proposed as an additional or alternative approach to standard risk assessment. The precautionary principle has also been advocated by some who see it as part of postmodern democracy in which more power is given to the public on health and safety matters relative to the judgments of technocrats. A more cynical view is that the precautionary principle is particularly championed by the European Community as a means to erect trade barriers. The WTO ruling against the European Community's trade barrier against beef from hormone-treated cattle seemed to support the use of risk assessment and appeared to reject the argument that the precautionary principle was a legitimate basis for trade barriers. However, a more recent WTO decision on asbestos contains language suggesting that the precautionary principle, in the form of taking into account public perception, may be acceptable as a basis for a trade barrier. This decision, if followed in future WTO trade disputes, such as for genetically modified foods, raises many issues central to the field of risk analysis. It is too early to tell whether the precautionary principle will become accepted in WTO decisions, either as a supplement or a substitute for standard risk assessment. But it would undermine the value of the precautionary principle if this principle were misused to justify unwarranted trade barriers. PMID: 15078320 [PubMed - indexed for MEDLINE] 1293: Toxicol Rev. 2003;22(2):83-90. Biopesticides. Sudakin DL. Department of Environmental and Molecular Toxicology, Oregon State University, Corvallis, Oregon, USA. sudakind@ace.orst.edu The term 'biopesticide' encompasses a broad array of microbial pesticides, biochemicals derived from micro-organisms and other natural sources, and processes involving the genetic incorporation of DNA into agricultural commodities that confer protection against pest damage (plant-incorporated protectants). Some microbial pesticides, such as Bacillus thuringiensis, have a long history of safe and effective use as a biological insecticide. More recent developments in microbial pest control include the utilisation of other bacterial and fungal species that may competitively inhibit the growth of pathogenic and toxigenic micro-organisms on important agricultural commodities. The use of microbes and their gene products introduces additional considerations to the toxicological dose-response relationship, including a need to determine the plausibility of infectious and immunological effects in association with human exposure to these biopesticides in food or the environment. Studies of substantial equivalence suggest that foods currently derived from plant-incorporated protectants are not likely to differ from conventional foods. However, there is general consensus that the scientific methods to assess risks from genetically modified foods and micro-organisms will continue to evolve in the future. Publication Types: Research Support, U.S. Gov't, Non-P.H.S. Review PMID: 15071818 [PubMed - indexed for MEDLINE] 1294: Transgenic Res. 2004 Feb;13(1):59-67. Peanut stripe potyvirus resistance in peanut (Arachis hypogaea L.) plants carrying viral coat protein gene sequences. Higgins CM, Hall RM, Mitter N, Cruickshank A, Dietzgen RG. Biotechnology, Department of Primary Industries, Queensland Agency for Food and Fibre Sciences, Queensland Bioscience Precinct, 306 Carmody Road, The University of Queensland, St. Lucia, Qld 4072, Australia. Peanut (Arachis hypogaea L.) lines exhibiting high levels of resistance to peanut stripe virus (PStV) were obtained following microprojectile bombardment of embryogenic callus derived from mature seeds. Fertile plants of the commercial cultivars Gajah and NC7 were regenerated following co-bombardment with the hygromycin resistance gene and one of two forms of the PStV coat protein (CP) gene, an untranslatable, full length sequence (CP2) or a translatable gene encoding a CP with an N-terminal truncation (CP4). High level resistance to PStV was observed for both transgenes when plants were challenged with the homologous virus isolate. The mechanism of resistance appears to be RNA-mediated, since plants carrying either the untranslatable CP2 or CP4 had no detectable protein expression, but were resistant or immune (no virus replication). Furthermore, highly resistant, but not susceptible CP2 T0 plants contained transgene-specific small RNAs. These plants now provide important germplasm for peanut breeding, particularly in countries where PStV is endemic and poses a major constraint to peanut production. Publication Types: Research Support, Non-U.S. Gov't PMID: 15070076 [PubMed - indexed for MEDLINE] 1295: Transgenic Res. 2004 Feb;13(1):5-19. Regulating transgenic crops: a comparative analysis of different regulatory processes. Jaffe G. Center for Science in the Public Interest, 1875 Connecticut Avenue, NW, Suite 300, Washington, DC 20009, USA. gjaffe@cspinet.org Transgenic crops have the potential to benefit both developed and developing countries. To ensure safe crops to humans and the environment, a strong, but not stifling, regulatory system needs to be established and properly implemented. This paper explores some essential components of a strong regulatory structure for transgenic crops. First, five different regulatory systems for transgenic crops--the United States, the European Union, South Africa, Taiwan, and Argentina--are described and explained. The major components of those systems are then compared to components necessary to a regulatory system that ensures safe products and engenders public trust. The key components discussed include: (1) mandatory pre-market approval; (2) established safety standards; (3) transparency; (4) public participation; (5) use of outside scientists for expert scientific advice; (6) independent agency decisions; (7) post-approval activities; and (8) enforcement authority and resources. Although no one of the existing systems analyzed adequately achieves all the necessary components of a strong regulatory system, those systems serve as models for deciding which regulatory procedures should be emulated and which should be avoided. A mandatory pre-market approval system that applies established safety standards in procedures that are transparent and allows for public participation with no pre-conceived notions or biases will best achieve both safe products and consumer trust. Publication Types: Comparative Study Review PMID: 15070071 [PubMed - indexed for MEDLINE] 1296: J Basic Microbiol. 2004;44(2):147-56. Microbial detoxification of mycotoxin deoxynivalenol. Völkl A, Vogler B, Schollenberger M, Karlovsky P. Max-Planck-Institut für Entwicklungsbiologie, Spemannstr. 35, D-72076 Tübingen, Germany. Deoxynivalenol (DON) is a trichothecene secondary metabolite produced by Fusarium species infecting cereal crops. As a mycotoxin, DON causes losses in livestock production and poses a health risk to humans consuming contaminated cereal products. DON also acts as a virulence factor, facilitating the colonization of host plants by Fusarium spp. Enzymatic detoxification of mycotoxins in feed additives and genetically modified crops is a promising approach for the reduction of mycotoxin contamination of feeds and food. A prerequisite for the development of biotechnological strategies for DON detoxification is the availability of genes encoding suitable enzymatic activities. With the goal of isolating microbial cultures that can be used as a source of such activities, we screened 1285 microbial cultures from farmland soil, cereal grains, insects and other sources for DON transformation under aerobic conditions. One mixed culture transformed DON into two chromatographically separable products. The main product of the transformation was purified and its structure was elucidated by mass spectroscopy, (1)H-NMR, (13)C-NMR and proton-proton and carbon-proton correlated NMR spectroscopy. The structure of this product was determined to be 3-keto-4-deoxynivalenol. The DON-transforming mixed culture survived and retained its transforming activity during a starvation period of six months at 20 degrees C. Transformation of DON was suppressed by low concentrations of glucose and high concentrations of tryptone and yeast extract. Cell-free supernatants obtained either by filtration through a 0.22 microm membrane filter or by centrifugation did not exert DON-transforming activity. Trichothecenes 15-acetyl-DON, 3-acetyl-DON and fusarenon-X were also transformed. Copyright 2004 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim Publication Types: Comparative Study Research Support, Non-U.S. Gov't PMID: 15069674 [PubMed - indexed for MEDLINE] 1297: Toxicol Lett. 2004 Apr 15;150(1):29-42. DNA microarray technology in nutraceutical and food safety. Liu-Stratton Y, Roy S, Sen CK. Laboratory of Molecular Medicine and DNA Microarray & Genetics Facility, Department of Surgery, Dorothy M. Davis Heart and Lung Research Institute, The Ohio State University Medical Center, 473 W. 12th Avenue, Columbus, OH 43210, USA. The quality and quantity of diet is a key determinant of health and disease. Molecular diagnostics may play a key role in food safety related to genetically modified foods, food-borne pathogens and novel nutraceuticals. Functional outcomes in biology are determined, for the most part, by net balance between sets of genes related to the specific outcome in question. The DNA microarray technology offers a new dimension of strength in molecular diagnostics by permitting the simultaneous analysis of large sets of genes. Automation of assay and novel bioinformatics tools make DNA microarrays a robust technology for diagnostics. Since its development a few years ago, this technology has been used for the applications of toxicogenomics, pharmacogenomics, cell biology, and clinical investigations addressing the prevention and intervention of diseases. Optimization of this technology to specifically address food safety is a vast resource that remains to be mined. Efforts to develop diagnostic custom arrays and simplified bioinformatics tools for field use are warranted. Publication Types: Review PMID: 15068823 [PubMed - indexed for MEDLINE] 1298: Proc Natl Acad Sci U S A. 2004 Apr 20;101(16):6315-20. Epub 2004 Apr 5. Enhanced translation of a chloroplast-expressed RbcS gene restores small subunit levels and photosynthesis in nuclear RbcS antisense plants. Dhingra A, Portis AR Jr, Daniell H. Department of Molecular Biology and Microbiology, University of Central Florida, 4000 Central Florida Boulevard, Biomolecular Science, Building 20, Room 336, Orlando, FL 32816-2364, USA. Ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) is a key enzyme that converts atmospheric carbon to food and supports life on this planet. Its low catalytic activity and specificity for oxygen leads to photorespiration, severely limiting photosynthesis and crop productivity. Consequently, Rubisco is a primary target for genetic engineering. Separate localization of the genes in the nuclear and chloroplast genomes and a complex assembly process resulting in a very low catalytic activity of hybrid Rubisco enzymes have rendered several earlier attempts of Rubisco engineering unsuccessful. Here we demonstrate that the RbcS gene, when integrated at a transcriptionally active spacer region of the chloroplast genome, in a nuclear RbcS antisense line and expressed under the regulation of heterologous (gene 10) or native (psbA) UTRs, results in the assembly of a functional holoenzyme and normal plant growth under ambient CO(2) conditions, fully shortcircuiting nuclear control of gene regulation. There was approximately 150-fold more RbcS transcript in chloroplast transgenic lines when compared with the nuclear RbcS antisense line, whereas the wild type has 7-fold more transcript. The small subunit protein levels in the gene 10/RbcS and psbA/RbcS plants were 60% and 106%, respectively, of the wild type. Photosynthesis of gene 10/RbcS plants was approximately double that of the antisense plants, whereas that of psbA/RbcS plants was restored almost completely to the wild-type rates. These results have opened an avenue for using chloroplast engineering for the evaluation of foreign Rubisco genes in planta that eventually can result in achieving efficient photosynthesis and increased crop productivity. Publication Types: Research Support, U.S. Gov't, Non-P.H.S. PMID: 15067115 [PubMed - indexed for MEDLINE] 1299: Eur Ann Allergy Clin Immunol. 2004 Feb;36(2):46-51. Severe food anaphylaxis: 107 cases registered in 2002 by the Allergy Vigilance Network. Moneret-Vautrin DA, Kanny G, Morisset M, Rancé F, Fardeau MF, Beaudouin E. Department of Internal Medicine, Clinical Immunology and Allergology, University Hospital, Avenue de Lattre de Tassigny, 54035 Nancy Cedex, France. BACKGROUND: The prevalence of food allergies increases, relating to diet modifications. The consumption of new foods--exotic foods or foods originally used for animal feed, new proteins, neo allergens due to the use of new technologies and soon, Genetically Modified Foods--are in the spotlight. OBJECTIVE: It is essential to develop a system of food allergy vigilance encompassing the full range of foods being consumed. Understanding this imperative leads logically to the suggestion of developing an allergy vigilance network taking advantage of the ongoing experience of allergists "on the ground". METHODS: The French Allergy Vigilance Network is subscribed to by 302 allergologists (267 of whom are French). The aims of the Network are to record cases of severe anaphylaxis, to establish an epidemiological data bank from prospective multicenter studies, and to monitor the allergic risk from novel foods. RESULTS: In 2002, 107 cases of severe anaphylaxis were recorded: anaphylactic shock--59.8% (one fatal), systemic reaction--18.7%, laryngeal angio-edema--15.9%, acute severe asthma--5.6% (one fatal). The main allergens identified were peanuts, nuts, shellfish, lupine flour and wheat flour. Action has been taken as a result: information by industry on inadequate labeling, withdrawal of wrongly labeled batches, and university hospital centers have been encouraged to establish the allergenic safety of their catering services. CONCLUSION: Setting up such a network in other countries would lead to a significant advance in knowledge of the peculiarities of allergies relating to a wide variety of eating habits. PMID: 15061394 [PubMed - indexed for MEDLINE] 1300: Plant Physiol Biochem. 2004 Jan;42(1):57-63. Transgenic tobacco plants accumulating osmolytes show reduced oxidative damage under freezing stress. Parvanova D, Ivanov S, Konstantinova T, Karanov E, Atanassov A, Tsvetkov T, Alexieva V, Djilianov D. Abiotic Stress Tolerance and Echophysiology, AgroBioInstitute, Institute of Cryobiology and Food Technology, Sofia, Bulgaria. We studied the reaction to the oxidative component of freezing in several tobacco lines, transformed with genes coding for enzymes involved in the synthesis of osmoprotectants (proline, fructan or glycine betaine) along with their wild type. The levels of some oxidative stress markers (leakage of electrolytes, hydrogen peroxide and malondialdehyde) as well as the activity of antioxidative enzymes catalase (EC 1.11.1.6.) and guaiacol peroxidase (EC 1.11.1.7.) have been followed at acclimation, 12 and 24 h freezing and at recovery. Freezing for 24 h resulted in severe damages for the wild type. A corresponding increase of electrolyte leakage, hydrogen peroxide and malondialdehyde contents, a rise of peroxidase activity and inhibition of catalase activity occurred in the non-transformants. Similar, but significantly lower trend of the same parameters has been found for the transgenic lines. Moreover, the oxidative markers returned to their normal levels when the transformants were able to recover from freezing. It could be speculated that transfer of genes, coding for accumulation of osmoprotectants, is related to reduced intensity of freezing-induced oxidative processes. Our lines and model system could serve as a good prerequisite for additional studies to gain further insights into the complex role of osmoprotectants in freezing tolerance. PMID: 15061085 [PubMed - indexed for MEDLINE] 1301: Nat Biotechnol. 2004 Apr;22(4):383-4. Flies in the soup--European GM labeling legislation. Craddock N. Neville Craddock Associates, Tun House, 28 High Street, Bletchingley, Surrey RH1 4PA, UK. nevillecraddock@tunhouse28.fsnet.co.uk PMID: 15060547 [PubMed - indexed for MEDLINE] 1302: Riv Biol. 2003 Sep-Dec;96(3):363-84. Transgenic foods as a tool for malnutrition elimination and their impact on agricultural systems. Monastra G, Rossi L. National Institute for Research on Food and Nutrition, Rome, Italy. monastra@inran.it GMO crops were introduced for commercial production in 1996. Since then, their use has increased rapidly. GMOs have primarily benefited large farms and multinational companies in Industrialised Countries and now is more and more debating their utilisation in Developing World. The objective of the present review is an analysis of this subject from a comprehensive point of view; in addition to that, the changes related to the nutritional content of transgenic foods will be treated. Despite the progress that has been made, the world food situation is still marked by mass hunger and chronic malnutrition. In particular micronutrient malnutrition, that means vitamin and mineral deficiencies, represents an important public health problem in several areas of the world. The "golden rice" bioengineered to contain beta-carotene, as a source of vitamin A is the most famous example of GM food used for reduction (or even to solve) of a public health problem. The expected results of this approach have presently not been achieved. Further studies are necessary to increase the general knowledge about GMOs and their long-term effects on human health. Collaborative attitude of different research sectors (private and public) and involvement of different sectors of society will be an added value for comprehension of the real impact of the application of modern biotechnology to food and agriculture systems. Publication Types: Review PMID: 15055878 [PubMed - indexed for MEDLINE] 1303: Nahrung. 2004 Feb;48(1):13-8. Quality and safety evaluation of genetically modified potatoes spunta with Cry V gene: compositional analysis, determination of some toxins, antinutrients compounds and feeding study in rats. El Sanhoty R, El-Rahman AA, Bögl KW. Federal Institute for Risk Assessment (BFR), Thielallee 88-92, D-14195 Berlin, Germany. r.sanhoty@bfr.bund.de The aim of this study was to evaluate the composition, nutritional and toxicology safety of GM potato Spunta lines compared to that of conventional potato Spunta. Compositional analyses were conducted to measure the proximate chemical composition with references to 14 components, total solid, protein, lipid, crude fibre, ash, carbohydrate, starch, reducing sugar, nonreducing sugar, sodium, calcium, potassium, phosphorus, and ascorbic acid. Some toxins and anti-nutrients compounds were determined. Feeding study of GM potatoes line (G2 and G3) in rats were done for 30 days. Four groups of albino rats were used for studying the effect and the safety assessment of GM potatoes Spunta G2 and G3. Group (I) was fed on control basal diet, group (II) was fed on control diet plus 30% freeze-dried nongenetically modified potato Spunta, group (III) was fed on control diet plus 30% freeze-dried genetically modified potato Spunta, and group (IV) was fed on control diet plus 30% freeze-dried genetically modified potato Spunta GMO G3. There were no significant differences between GM potatoes G2, G3, and Spunta control potato line in the proximate chemical composition. The levels of glycoalkaloids in transgenic potato tubers and nontransgenic were determined and there were also no significant differences between the GM potatoes and conventional potato line, the levels were in agreement with a safety level recommended by FAO/WHO (200 mg/ kg) for acute toxicity. Protease inhibitor activity and total phenol were estimated and no significant differences between the GM potatoes line and conventional potato Spunta line were found. During the period tested, rats in each group (I, II, III, IV) grew well without marked differences in appearance. No statistical difference were found in food intake, daily body weight gain and feed efficiency. But there is a slightly significant difference in finally body weight between the control group and experimental groups. No significant difference were found in serum biochemical value between each groups, and also between relative organs weight (liver, spleen, heart, kidney, testes). From these results, it can be concluded that the GM potatoes Spunta line (G2 and G3) with Cry V gene are confirmed to have nearly the composition and biochemical characteristics as non-GM potato Spunta. PMID: 15053345 [PubMed - indexed for MEDLINE] 1304: Rocz Panstw Zakl Hig. 2003;54(4):345-53. [Detection of genetic modification in maize and maize products by ELISA-test] [Article in Polish] Urbanek-Karłowska B, Sawilska-Rautenstrauch D, Jedra M, Badowski P. Zakład Badania Zywności i Przedmiotów Uzytku, Państwowy Zakład Higieny, 00-791 Warszawa, ul. Chocimska 24. Enzyme immunoassay methods--TRAIT Test--was applied for detection of genetic modification in maize seeds and foodstuffs, which have been produced from this crop. TRAIT Test is based on the identification GMO protein Cry 1Ab produced by a gene derived from Bacillus thuringiensis (Bt) incorporated into insect resistant corn grain. The experiment was carried out on maize standards and foodstuffs from Warsaw market. The positive result was obtained for one maize product, which was not labelled as GMO. The presence of GMO material was approximately equal to 1%. In conclusion, this test is proper for fast routine qualitative (yes/no) determination GMO material in maize seeds and unprocessed food products. Publication Types: English Abstract PMID: 15052732 [PubMed - indexed for MEDLINE] 1305: Proc Natl Acad Sci U S A. 2004 Apr 6;101(14):5158-63. Epub 2004 Mar 24. Enhancement of folates in plants through metabolic engineering. Hossain T, Rosenberg I, Selhub J, Kishore G, Beachy R, Schubert K. Donald Danforth Plant Science Center, 975 North Warson Road, St. Louis, MO 63132, USA. Humans depend on plants as a major source of dietary folates. Inadequate dietary levels of the vitamin folate can lead to megaloblastic anemia, birth defects, impaired cognitive development, and increased risk of cardiovascular disease and cancer. The biofortification of folate levels in food crops is a target for metabolic engineering. Folates are synthesized de novo from pterins and para-amino benzoic acid, which are subsequently combined to form dihydropteroate, the direct precursor to dihydrofolate. We postulated that GTP cyclohydrolase-1, which catalyzes the first committed step in pterin biosynthesis, was a rate-limiting step in pterin synthesis in plants and, therefore, in folate synthesis. On this basis, we proposed that the expression of an unregulated bacterial GTP cyclohydrolase-1 in plants would increase pterin biosynthesis with a concomitant enhancement of folate levels. The folE gene encoding GTP cyclohydrolase-1 was cloned from Escherichia coli and introduced into Arabidopsis thaliana through plant transformation. The expression of bacterial GTP cyclohydrolase-1 in transgenic Arabidopsis resulted in a 1,250-fold and 2- to 4-fold enhancement of pterins and folates, respectively. These results helped to identify other potential factors regulating folate synthesis, suggesting ways to further enhance folate levels in food crops. Publication Types: Research Support, U.S. Gov't, Non-P.H.S. PMID: 15044686 [PubMed - indexed for MEDLINE] 1306: Trends Biotechnol. 2004 Mar;22(3):107-9. Comment in: Trends Biotechnol. 2005 Aug;23(8):386-7. How can genetically modified foods be made publicly acceptable? Rowe G. Institute of Food Research, Norwich Research Park, Colney, Norwich NR4 7UA, UK. gene.rowe@bbsrc.ac.uk A recent study by Lusk suggests that consumers might voluntarily pay more for a genetically modified (GM) food than a non-GM equivalent if made aware of the possible health benefits. However, other research indicates that the acceptability of novel hazards is affected by a variety of factors, in addition to benefits, and that making agricultural biotechnology publicly acceptable will be more complex than indicated by the results from Lusk's study. Publication Types: Research Support, Non-U.S. Gov't PMID: 15043045 [PubMed - indexed for MEDLINE] 1307: Nat Rev Drug Discov. 2004 Feb;3(2):152-9. Oxygen therapeutics: can we tame haemoglobin? Alayash AI. Laboratory of Biochemistry, Division of Hematology, Center for Biologics Evaluation and Research, Food and Drug Administration, 8800 Rockville Pike, National Institutes of Health, Bethesda, Maryland 20892, USA. alayash@cber.fda.gov Chemically modified or genetically engineered haemoglobins (Hbs) developed as oxygen therapeutics (often termed 'blood substitutes') are designed to correct oxygen deficit due to ischaemia in a variety of clinical settings. These modifications are intended to stabilize Hb outside its natural environment--red blood cells--in a functional tetrameric and/or polymeric form. Uncontrolled haem-mediated oxidative reactions of cell-free Hb and its reactions with various oxidant/antioxidant and cell signalling systems have emerged as an important pathway of toxicity. Current protective strategies designed to produce safe Hb-based products are focused on controlling or suppressing the 'radical' nature of Hb while retaining its oxygen-carrying function. Publication Types: Review PMID: 15043006 [PubMed - indexed for MEDLINE] 1308: Shokuhin Eiseigaku Zasshi. 2003 Dec;44(6):281-8. [Laboratory-performance study of the notified methods to detect genetically modified maize (CBH351) and potato (NewLeaf Plus and NewLeaf Y)] [Article in Japanese] Watanabe T, Kasama K, Wakui C, Shibuya M, Matsuki A, Akiyama H, Maitani T. National Institute of Health Sciences: 1-18-1, Kamiyoga, Setagaya-ku, Tokyo 158-8501, Japan. To investigate the key factors affecting the reliability of the analytical results, a laboratory-performance study was attempted for the notified methods to detect genetically modified (GM) maize (CBH351) and GM potato (NewLeaf Plus and NewLeaf Y). The test samples were designed as three pairs of blind duplicates, which included 0%, 0.1% and 1.0% GM maize (CBH351) or GM potato (NewLeaf Plus or NewLeaf Y). Fourteen laboratories participated in the study. The test samples were sent to the participating laboratories along with the protocol. The data were collected from all laboratories and statistically analyzed. For the 0% sample of the CBH351 maize, one laboratory reported a false-positive result. It was considered that contamination could have occurred via the common use of equipment or tools for the test. For the 0.1% samples of the NewLeaf Plus potato or NewLeaf Y potato, on the other hand, three laboratories reported false-negative results. It was presumed that these results were due to changes of the conditions of the electrophoresis and agarose-gel staining. The other laboratories reported appropriate results. It was considered that the method employed in this study was suitable for the assessment of laboratory performance. Publication Types: English Abstract PMID: 15038109 [PubMed - indexed for MEDLINE] 1309: Exp Gerontol. 2004 Feb;39(2):269-72. A transgenic dwarf rat model as a tool for the study of calorie restriction and aging. Yamaza H, Komatsu T, Chiba T, Toyama H, To K, Higami Y, Shimokawa I. Department of Pathology and Gerontology, Nagasaki University Graduate School of Biomedical Sciences, 1-12-4 Sakamoto, Nagasaki City 852-8523, Japan. We have previously reported a long-lived transgenic dwarf rat model, in which the growth hormone (GH)-insulin like growth factor (IGF)-1 axis was selectively suppressed by overexpression of antisense GH transgene. Rats heterozygous for the transgene (tg/-) manifest phenotypes similar to those in calorie-restricted (CR) rats. To further characterize the transgenic rat in comparison with CR rats, the present study evaluated glucose and insulin tolerance in tg/- and control Wistar (-/-) rats at 6-9 months of age. Rats were fed ad libitum (AL) or 30% CR from 6 weeks of age. In CR rats, glucose disposal after glucose load was facilitated without any significant surge of serum insulin, and insulin tolerance test also indicated increased insulin sensitivity. In transgenic rats, similar findings were observed after glucose and insulin load, and CR in tg/- rats further facilitated glucose disposal during glucose and insulin tolerance tests. These findings suggest the presence of both common and separate mechanisms regulating the glucose-insulin system between CR and the reduced GH-IGF-1 axis paradigms. The transgenic rat model is, therefore, a useful one for studies of CR and aging. Publication Types: Research Support, Non-U.S. Gov't PMID: 15036422 [PubMed - indexed for MEDLINE] 1310: Law Hum Genome Rev. 2003 Jul-Dec;(19):159-85. [The zig-zagging legislative policies of the European Union in relation to transgenic products] [Article in Spanish] Martín Uranga A. Universidad de Deusto-Universidad del País Vasco/EHU, Madrid, España. The author analyses the E.U. legislative policies related to transgenic products from the 80s until nowadays, she stops in Directive 2001/18/Ce and its conversion into national law. She explains how the regulatory frame is after the recent approval of the newest rules about tracking and labelling genetically modified organisms (OMG). She also studies communitary jurisprudence about this subject according to the legislation, concretely sentence 9 September 2003 in the affair C-236/01 about commercialisation of two lines of transgenic corn. Finally she points out how legislative policies have influenced the creation and development of biotechnological enterprises. Publication Types: English Abstract Review PMID: 15032103 [PubMed - indexed for MEDLINE] 1311: Risk Anal. 2004 Feb;24(1):185-94. GM foods and the misperception of risk perception. Gaskell G, Allum N, Wagner W, Kronberger N, Torgersen H, Hampel J, Bardes J. Department of Social Psychology, London School of Economics, Centre for the Analysis of Risk and Regulation, London, UK. g.gaskell@lse.ac.uk Public opposition to genetically modified (GM) food and crops is widely interpreted as the result of the public's misperception of the risks. With scientific assessment pointing to no unique risks from GM crops and foods, a strategy of accurate risk communication from trusted sources has been advocated. This is based on the assumption that the benefits of GM crops and foods are self-evident. Informed by the interpretation of some qualitative interviews with lay people, we use data from the Eurobarometer survey on biotechnology to explore the hypothesis that it is not so much the perception of risks as the absence of benefits that is the basis of the widespread rejection of GM foods and crops by the European public. Some respondents perceive both risks and benefits, and may be trading off these attributes along the lines of a rational choice model. However, for others, one attribute-benefit-appears to dominate their judgments: the lexicographic heuristic. For these respondents, their perception of risk is of limited importance in the formation of attitudes toward GM food and crops. The implication is that the absence of perceived benefits from GM foods and crops calls into question the relevance of risk communication strategies for bringing about change in public opinion. PMID: 15028010 [PubMed - indexed for MEDLINE] 1312: Asia Pac J Clin Nutr. 2003;12 Suppl:S31. Modern pork production - Balancing efficient growth and feed conversion with product quality requirements and consumer demands. Van Barneveld RJ. Barneveld Nutrition Pty Ltd, South Maclean, QLD 4280. Background - Profitable pork production in an environment of increasing global competitiveness, diverse markets and heightened consumer awareness is an ongoing challenge. The modern pork producer needs to balance the use of emerging technologies and intensive farming practices, which have the potential to significantly decrease the cost of production, with specific market requirements for a quality product. Manipulation of fat content and distribution, nutritional enrichment of pork products, use of 'clean and green' feed ingredients, and hormone and antibiotic free production practices are high on the list of requirements of most markets as is the relative cost of Australian pork to imports and other meats. As some of these requirements are conflicting, modern pork producers must adopt some innovative nutritional and production strategies if they are to remain commercially viable. Review - In a modern Australian pig production system, a sale liveweight of 96 kg for the domestic market can be achieved in approximately 160 days with a feed conversion ratio of 2.60:1. This level of efficiency has been achieved through intense genetic selection for fast growing, lean animals over the past 30 years, an advanced knowledge of the nutritional value of feed ingredients and the nutritional requirements of pigs, and sophisticated production tools including advanced growth simulation models such as AUSPIG. Consumer demands for reduced levels of fat in pork has contributed to this efficiency given the high energetic cost of fat deposition relative to lean meat deposition. The development of export markets for Australian pork in 1996 coupled with a concurrent increase in consumer awareness of food production has prompted a renewed focus on factors influencing pork product quality traits in addition to the cost of production, particularly manipulation of fat quality, fat composition and fat distribution. For example, higher levels of intramuscular fat or marbling in pork, largely influenced by genetics, can positively affect the juiciness, tenderness and flavour of pork. In addition, a focus on dietary fat sources for growing pigs has been shown to influence fat quality and composition. Restricting the level of dietary unsaturated fats can reduce the incidence of soft fat, and restricting the use of dietary fish oils to 0.5% for at least two weeks prior to slaughter will maintain the processing and keeping qualities of the pork. In contrast, modified forms of fishmeal as a dietary source of long chain n-3 PUFA can effectively be used to produce n-3 enriched pork, a factor with potential to greatly increase the attractiveness of pork to some markets. As well as manipulation of the fat attributes of pork, other nutritional strategies that have potential to improve the quality of pork products include enrichment using dietary supplements of minerals such as selenium and a reduced reliance on antibiotics through improved nutrition and herd health status. Conclusions - Modern pork production involves rapid responses to changing market demands. A major issue faced by Australian producers is the need balance product quality requirements with increased costs of production. This is further confounded by the fact that modern technologies such as immunocastration, and the use of porcine somatotropin and genetically modified feed ingredients, all of which could potentially further enhance product quality while offsetting some of the costs of production, are poorly accepted in the market place. PMID: 15023634 [PubMed - in process] 1313: Proc Nutr Soc. 2003 Nov;62(4):901-12. Impact of genetic variation on metabolic response of bone to diet. Cusack S, Cashman KD. Department of Food and Nutritional Sciences, University College, Cork, Republic of Ireland. There is compelling evidence to suggest that both the development of bone to peak bone mass at maturity and subsequent loss depend on the interaction between genetic, hormonal, environmental and nutritional factors. The major part (< or = 80%) of the age-specific variation in bone turnover and bone density is genetically determined. However, the notion of genetic determinant is of little value unless the specific genes that are involved can be identified. Most work in this area of osteoporosis research has focused on the candidate gene approach, which has identified several candidate genes for osteoporosis, including genes encoding the vitamin D receptor (VDR), oestrogen receptors (alpha and beta), apolipoprotein E, collagen type I alpha 1 and methylenetetrahydrofolate reductase, amongst many others. However, in general, findings from numerous studies of the association between such genes and various bone variables have been inconsistent. In addition to possible gene-gene interactions it is likely that there are interactions between these genes and certain environmental factors, especially nutrition, that may mediate expression of bone-related phenotypes. While these potential interactions add a level of complexity to our understanding of these apparent genetic effects on bone, identification of a role for genetic factors without knowledge of their interaction with nutrients can do little to advance prevention and treatment of osteoporosis. This information is especially important because, unlike genotype, diet and nutrition can be modified. The aim of the present review is to critically evaluate current knowledge relating to candidate genes for osteoporosis, with particular emphasis on their interaction with nutrients and dietary factors in determining bone health. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 15018490 [PubMed - indexed for MEDLINE] 1314: Science. 2004 Mar 12;303(5664):1590. Transgenic crops. Britain opts for brave new GM world. Vogel G. Publication Types: News PMID: 15016966 [PubMed - indexed for MEDLINE] 1315: Nature. 2004 Mar 11;428(6979):107. Californian county bans transgenic crops. Dalton R. Publication Types: News PMID: 15014455 [PubMed - indexed for MEDLINE] 1316: Nature. 2004 Mar 11;428(6979):107. Transgenic planting approved despite scepticism of UK public. Giles J. Publication Types: News PMID: 15014454 [PubMed - indexed for MEDLINE] 1317: Mutat Res. 2004 Mar 22;547(1-2):5-18. Analysis of mutations and bone marrow micronuclei in Big Blue rats fed leucomalachite green. Manjanatha MG, Shelton SD, Bishop M, Shaddock JG, Dobrovolsky VN, Heflich RH, Webb PJ, Blankenship LR, Beland FA, Greenlees KJ, Culp SJ. Food and Drug Administration, National Center for Toxicological Research, 3900 NCTR Road, Jefferson, AR 72079, USA. Leucomalachite green (LMG) is the major metabolite of malachite green (MG), a triphenylmethane dye that has been used widely as an antifungal agent in the fish industry. Concern over MG and LMG is due to the potential for consumer exposure, suggestive evidence of tumor promotion in rodent liver, and suspicion of carcinogenicity based on structure-activity relationships. In order to evaluate the risks associated with exposure to LMG, female Big Blue rats were fed up to 543 ppm LMG; groups of these rats were killed after 4, 16, or 32 weeks of exposure and evaluated for genotoxicity. We previously reported that this treatment resulted in a dose-dependent induction of liver DNA adducts, and that the liver lacI mutant frequency (MF) was increased, but only in rats fed 543 ppm LMG for 16 weeks. In the present study, we report the results from lymphocyte Hprt mutant assays and bone marrow micronucleus assays performed on these same rats. In addition, we have determined the types of lacI mutations induced in the rats fed 543 ppm LMG for 16 weeks and the rats fed control diet. No significant increases in the frequency of micronuclei or Hprt mutants were observed for any of the doses or time points assayed. Molecular analysis of 80 liver lacI mutants from rats fed 543 ppm LMG for 16 weeks revealed that 21% (17/80) were clonal in origin and that most (55/63) of the independent mutations were base pair substitutions. The predominant type of mutation was G:C --> A:T transition (31/63) and the majority (68%) of these involved CpG sites. When corrected for clonality, the 16-week lacI mutation frequency (36 +/- 10) x 10(-6) in treated rats was not significantly different from the clonally corrected control frequency (17 +/- 9 x 10(-6); P = 0.06). Furthermore, the lacI mutational spectrum in treated rats was not significantly different from that found for control rats (P = 0.09). Taken together, these data indicate that the DNA adducts produced by LMG in female rats do not result in detectable levels of genotoxicity, and that the increase in lacI MF observed previously in the liver of treated rats may be due to the disproportionate expansion of spontaneous lacI mutations. Publication Types: Comparative Study Research Support, U.S. Gov't, Non-P.H.S. PMID: 15013694 [PubMed - indexed for MEDLINE] 1318: Formos J Med Humanit. 2001 Oct;2(1-2):41-7. New technology ethic. Macer D. Eubios Ethics Institute; Institute of Biological Sciences, University of Tsukuba, Tsukuba, Japan. PMID: 15011655 [PubMed - indexed for MEDLINE] 1319: Peptides. 2004 Jan;25(1):37-43. Optimal designing of beta-conglycinin to genetically incorporate RPLKPW, a potent anti-hypertensive peptide. Onishi K, Matoba N, Yamada Y, Doyama N, Maruyama N, Utsumi S, Yoshikawa M. Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Gokasho Uji, Kyoto 611-0011, Japan. Previously, we introduced the RPLKPW sequence, a highly potent hypotensive peptide designed based on ovokinin (2-7), into three homologous sites in the soybean beta-conglycinin alpha' subunit by site-directed mutagenesis. The modified protein expressed in Escherichia coli reduced blood pressure of spontaneously hypertensive rats (SHRs) after oral administration at a dose of 10 mg/kg, which suggested about 30% of the introduced peptide was released in vivo. In this study amino acid residues around the RPLKPW sequence were optimized with a use of synthetic peptides to facilitate release of RPLKPW by gastrointestinal proteases. Then, fourth RPLKPW was also introduced into the extension domain of the protein. The newly modified protein, which was produced in E. coli, significantly lowered blood pressure in SHRs at a dose of 2.5 mg/kg 4 h after oral administration. Furthermore, we produced an extension domain that corresponds to residues 1-143 of the modified alpha' subunit containing four RPLKPW sequences by introducing a termination codon. The minimum effective dose of the modified extension domain was 1.0 mg/kg, which is 1/2000 that of ovalbumin. Publication Types: Research Support, Non-U.S. Gov't PMID: 15003354 [PubMed - indexed for MEDLINE] 1320: Curr Opin Plant Biol. 2004 Apr;7(2):210-8. Improving starch for food and industrial applications. Jobling S. s.jobling1@ntlworld.com Progress in understanding starch biosynthesis, and the isolation of many of the genes involved in this process, has enabled the genetic modification of crops in a rational manner to produce novel starches with improved functionality. For example, potato starches have been created that contain unprecedented levels of amylose and phosphate. Amylose-free short-chain amylopectin starches have also been developed; these starches have excellent freeze-thaw stability without the need for chemical modification. These developments highlight the potential to create even more modified starches in the future. Publication Types: Review PMID: 15003223 [PubMed - indexed for MEDLINE] 1321: Curr Opin Plant Biol. 2004 Apr;7(2):189-95. Safe and acceptable strategies for producing foreign molecules in plants. Mascia PN, Flavell RB. Ceres Inc., 3007 Malibu Canyon Road, Malibu, California 90265, USA. pmascia@ceres-inc.com The ability to express foreign genes using transgenic technologies has opened up options for producing large quantities of commercially important industrial or pharmaceutical products in plants. These technologies have made it possible to use well-developed systems of commercial agriculture that were developed principally to produce raw material for large-scale food, feed or processing applications for the production of foreign molecules. The possibility of the novel industrial or pharmaceutical molecules produced in such plants, or components derived from them, contaminating the environment and food chains has become especially controversial. This potential contamination has prompted detailed consideration of how such crops and the molecules that they produce can be effectively isolated and contained. First, the crop can be completely isolated physically from its food or feed counterpart during every aspect of its development and commercialization. Second, genetic isolation systems or genetic barriers that prevent normal reproduction can be used to reduce the likelihood of the industrial or pharmaceutical crop entering the food chain. Publication Types: Review PMID: 15003220 [PubMed - indexed for MEDLINE] 1322: Nature. 2004 Mar 4;428(6978):6. US and biologists wary of strict biotech rules. Cyranoski D. Publication Types: News PMID: 14999250 [PubMed - indexed for MEDLINE] 1323: JAMA. 2004 Mar 3;291(9):1055. Prevent genetically modified organisms from escaping into nature, report urges. Hampton T. Publication Types: News PMID: 14996758 [PubMed - indexed for MEDLINE] 1324: J Agric Food Chem. 2004 Mar 10;52(5):1390-7. Lower fumonisin mycotoxin levels in the grain of Bt corn grown in the United States in 2000-2002. Hammond BG, Campbell KW, Pilcher CD, Degooyer TA, Robinson AE, McMillen BL, Spangler SM, Riordan SG, Rice LG, Richard JL. Product Safety Center, Monsanto Company, 800 North Lindbergh Boulevard, St. Louis, Missouri 63167, USA. bruce.g.hammond@monsanto.com Fumonisins were monitored in corn grain collected from Bt hybrids grown in 107 locations across the United States in 2000-2002. Bt corn hybrids contain the Cry1Ab protein from Bacillus thuringiensis that controls European corn borers and other stalk-boring pests. Fumonisin levels were frequently lower in grain from Bt hybrids grown in field trials under conditions of natural (FACT trials) or manual insect infestation (university trials). Over three years of FACT trials, there were 126/210 comparisons when fumonisin levels in grain from control hybrids were >2 ppm, exceeding U.S. FDA guidance levels of 2 ppm for human food. Grain from Bt hybrids was at or below 2 ppm of fumonisins for 58 of the 126 comparisons. The use of Bt hybrids can increase the percentage of corn grain that would be suitable for use in food and feed. PMID: 14995151 [PubMed - indexed for MEDLINE] 1325: J Agric Food Chem. 2004 Mar 10;52(5):1049-54. Detection and quantitation of genetically modified maize (Bt-176 transgenic maize) by applying ligation detection reaction and universal array technology. Bordoni R, Mezzelani A, Consolandi C, Frosini A, Rizzi E, Castiglioni B, Salati C, Marmiroli N, Marchelli R, Rossi Bernardi L, Battaglia C, De Bellis G. Consiglio Nazionale delle Ricerche, Istituto di Tecnologie Biomediche and Dipartimento di Scienze e Tecnologie Biomediche, Università degli Studi di Milano, LITA, Via F.lli Cervi, 93 20090 Segrate, Milan, Italy. We have applied the ligation detection reaction (LDR) combined with a universal array approach to the detection and quantitation of the polymerase chain reaction (PCR) amplified cry1A(b) gene from Bt-176 transgenic maize. We demonstrated excellent specificity and high sensitivity. Down to 0.5 fmol (nearly 60 pg) of PCR amplified transgenic material was unequivocally detected with excellent linearity within the 0.1-2.0% range with respect to wild-type maize. We suggest the feasibility of extending the LDR/universal array format to detect in parallel several transgenic sequences that are being developed for food applications. Publication Types: Research Support, Non-U.S. Gov't PMID: 14995096 [PubMed - indexed for MEDLINE] 1326: J Relig Ethics. 2003 Winter;31(3):363-98. Bioethics, theology, and social change. Cahill LS. Recent years have witnessed a concern among theological bioethicists that secular debate has grown increasingly "thin," and that "thick" religious traditions and their spokespersons have been correspondingly excluded. This essay disputes that analysis. First, religious and theological voices compete for public attention and effectiveness with the equally "thick" cultural traditions of modern science and market capitalism. The distinctive contribution of religion should be to emphasize social justice in access to the benefits of health care, challenging the for-profit global marketing of research and biotechnology to wealthy consumers. Second, religion and theology have been and are still socially effective in sponsoring activism for practical change, both locally and globally. This claim will be supported with specific examples; with familiar concepts like subsidiarity and "middle axioms"; and with recent analyses of "participatory democracy" and of emerging, decentralized forms of global governance. PMID: 14986639 [PubMed - indexed for MEDLINE] 1327: FDA Consum. 2003 Nov-Dec;37(6):28-34. Genetic engineering: the future of foods? Bren L. PMID: 14986586 [PubMed - indexed for MEDLINE] 1328: Plant Cell Rep. 2004 Jul;22(12):894-902. Epub 2004 Feb 25. Transgenic Pinus radiata from Agrobacterium tumefaciens-mediated transformation of cotyledons. Grant JE, Cooper PA, Dale TM. New Zealand Institute for Crop and Food Research Limited, Private Bag 4704, Christchurch. grantj@crop.cri.nz A method for Agrobacterium tumefaciens-mediated transformation of Pinus radiata cotyledon explants was developed using commercially available open-pollinated seed. Pinus radiata is the most widely planted commercial conifer species in the Southern Hemisphere. Reports on transformation of this species have relied on particle bombardment of embryogenic callus derived from immature embryos. The main drawback to the method is the small number of genotypes that are amenable to transformation and regeneration. Since more than 80% of genotypes of radiata pine can be regenerated using cotyledons from mature seed, cotyledon explants were cocultivated with A. tumefaciens strain AGL1 containing a plasmid coding for the neomycin phosphotransferase II (nptII) gene and the beta-glucuronidase (GUS) gene (uidA). Transformed shoots were selected using either geneticin or kanamycin. Critical factors for successful transformation were survival of the cotyledons after cocultivation and selection parameters. Of the 105 putative transformants that were recovered from selection media, 70% were positive for integration of the nptII gene when analysed by PCR. GUS histochemical assay for uidA expression was unreliable because of reaction inhibition by unidentified compounds in the pine needles. Further, only 4 of the 26 independent transformants characterised by PCR and Southern analysis contained an intact copy of both genes. The remaining 22 transformants appeared to have a truncated or rearranged copy of the T-DNA. It is possible that the truncation/rearrangements are due to the Cauliflower mosaic virus (CaMV) 35S promoter. Analysis of the T-DNA junction sites and sequencing of the introduced DNA will help elucidate the nature of T-DNA insertion so that genetic modification of radiata pine can be targeted effectively. Publication Types: Research Support, Non-U.S. Gov't PMID: 14986058 [PubMed - indexed for MEDLINE] 1329: J Econ Entomol. 2003 Dec;96(6):1738-49. Use of transgenic plants to measure insect herbivore movement. Spencer JL, Mabry TR, Vaughn TT. Center for Economic Entomology, Illinois Natural History Survey, 607 E. Peabody Drive, Champaign, IL 61820, USA. spencer1@uiuc.edu Use of ingested transgenic corn tissue as a marker for measuring movement of adult Diabrotica virgifera virgifera (LeConte) (Coleoptera: Chrysomelidae; western corn rootworm) was investigated. Laboratory observations of beetles feeding on corn foliage, pollen, silks, or soybean foliage provided background on feeding patterns. The interval between food consumption and its appearance in feces (gut passage time) ranged from 102.7 +/- 11 min for soybean foliage to 56.7 +/- 2.9 min for corn silks. In a laboratory assay, protein expression tests identified the presence of Cry3Bb1 protein inside 50% of adult D. virgifera for up to 16 h after they had last consumed Cry3Bb1 protein-expressing corn silks from 'YieldGard Rootworm' corn plants (Monsanto Co.). Cry3Bb1 protein could not be detected by 32 h postfeeding. The proportion of Cry3Bb1 protein-positive beetles declined linearly with increasing time since feeding on 'YieldGard Rootworm' tissue. Approximately 20% of adult D. virgifera collected near 'YieldGard Rootworm' corn plots tested positive for Cry3Bb1 protein, indicating 'YieldGard Rootworm' tissue consumption within the last 16-32 h. Based on a 16- to 32-h postfeeding detection interval for Cry3Bb1 protein and the distance between 'YieldGard Rootworm' sources and sites where Cry3Bb1-positive insects were collected, 85.3% of males and females moved < or = 4.6-9.1 m/d through R2-R3 stage corn. Among Cry3Bb1-positive adults that left corn and were captured in an adjacent soybean field, 86.4% of males and 93.1% of females moved < or = 4.6-9.1 m/d through soybean. Detection of transgenic plant tissues in mobile insect herbivores is a novel application of biotechnology to the study of insect movement. Publication Types: Research Support, Non-U.S. Gov't PMID: 14977111 [PubMed - indexed for MEDLINE] 1330: Pest Manag Sci. 2004 Feb;60(2):105-12. Crop protection--what will shape the future picture? Atkinson D, Litterick AM, Walker KC, Walker R, Watson CA. Scottish Agricultural College, West Mains Road, Edinburgh EH9 3JG, UK. D.Atkinson@ed.sac.ac.uk The drivers which influence the types of crop protection most needed by agriculture are changing. A polarisation of approaches has resulted in the needs of organic agriculture and biotechnological agriculture, which can be identified as the current extremes of the spectrum, becoming very different. The main requirements of these two sectors are identified here and used as the basis for questioning future EU research requirements in crop protection. Factors affecting/influencing organic farming and the use of genetically modified crops are discussed in some detail. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 14971676 [PubMed - indexed for MEDLINE] 1331: J Agric Food Chem. 2004 Feb 25;52(4):809-15. Method for the detection of synthetic cry3A in transgenic potatoes. Smith DS, Maxwell PW, De Boer SH. Centre for Animal and Plant Health, Canadian Food Inspection Agency, 93 Mount Edward Road, Charlottetown, Prince Edward Island C1A 5T1, Canada. smithds@inspection.gc.ca All transgenic cultivars of potatoes registered in Canada and the United States have been modified to express a synthetic cry3A gene as a means of conferring resistance against the Colorado potato beetle, an important economic pest of potatoes. A PCR method was developed to amplify a 499 bp region of the synthetic cry3A gene. Using this method, synthetic cry3A could be detected in six different transgenic cultivars. Positive results could be confirmed with PvuII restriction digestion of the PCR-generated amplicon, which resulted in two fragments that were 283 and 216 bp in size. Of the 52 tuber extracts tested with this method, no false positive or false negative results were obtained, suggesting the method could be used with a high degree of accuracy. The absolute limit of detection was the number of cry3A copies present in one or perhaps two haploid copies of the potato genome. The practical limit of detection in tubers on a fresh weight basis was 0.02% for the NL 10-SUP and 0.01% for the remaining cultivars. Synthetic cry3A could also be detected in processed food products such as potato chips, shoestring potatoes, and frozen French fries. The method was suitable for screening potato tuber lots and some processed foods for the presence of synthetic cry3A. PMID: 14969535 [PubMed - indexed for MEDLINE] 1332: Methods. 2004 Mar;32(3):340-5. Knocking out expression of plant allergen genes. Bhalla PL, Singh MB. Plant Molecular Biology and Biotechnology Laboratory, ARC Centre of Excellence for Integrative Legume Research, Institute of Land and Food Resources, The University of Melbourne, Parkville, Vic 3010, Australia. premlb@unimelb.edu.au Pollen of many grasses, trees, and weeds are the source of inhalant allergic proteins while various other plant products are allergenic only upon their ingestion as a food source. Allergenic proteins of pollen are exposed to human immune system after their rapid release from pollen upon coming in contact with moist surface of nasal mucosa. The advent of molecular cloning and ability to genetically transform plants now offer unprecedented opportunities to produce hypoallergenic plants by targeted switching off allergen production. Gene silencing strategies that operate at post-transcriptional level are highly suitable for blocking allergen production. We have demonstrated the concept of allergen gene silencing through antisense approach by producing ryegrass plants that do not produce major allergen in its pollen. Our results show the potential of antisense approach in reducing the allergenic potential of plants. Such a strategy can have a general applicability for production of transgenic plants depleted of both inhaled and ingested allergens. In addition, such an approach could also help in elucidating the in vivo function of allergen(s) in plants and contribution of an allergen to overall allergenic potential of an allergen source. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 14962769 [PubMed - indexed for MEDLINE] 1333: APMIS. 2004 Jan;112(1):21-8. A comparative study of the allergenic potency of wild-type and glyphosate-tolerant gene-modified soybean cultivars. Sten E, Skov PS, Andersen SB, Torp AM, Olesen A, Bindslev-Jensen U, Poulsen LK, Bindslev-Jensen C. Allergy Clinic, 7542, National University Hospital, Blegdamsvej 9, 2100 Copenhagen Ø, Denmark. E.STEN@RH.DK A large proportion of soybean cultivars grown in the USA are now genetically modified varieties and concern has been raised about the safety of these products for consumers. A study of the impact on allergenic potency in soybeans, comparable except for the newly introduced gene (CP4 EPSPS), was performed using soybean-sensitized patients. The allergenicity of 18 different (10 GM and 8 WT) soybean extracts was examined blindly by the following three methods: A) Sera from patients with specific IgE against soybean were used to determine concentrations inducing 50% RAST inhibition; B) Histamine release induced by the extracts was examined using blood from sensitized patients; C) SPT was performed on sensitized patients with all 18 extracts. All three methods showed variations in the allergenic potency between the individual extracts but allergenic potential was not affected by presence of the transgene. By using standard in vitro methods and SPT for determination of allergenicity we were not able to detect any significant difference in the allergenic potency between GM and WT soybeans. Publication Types: Comparative Study Research Support, Non-U.S. Gov't PMID: 14961970 [PubMed - indexed for MEDLINE] 1334: Food Chem Toxicol. 2004 Mar;42(3):493-502. Persistence of DNA studied in different ex vivo and in vivo rat models simulating the human gut situation. Wilcks A, van Hoek AH, Joosten RG, Jacobsen BB, Aarts HJ. Institute of Food Safety and Nutrition, Danish Veterinary and Food Administration, Mørkhøj Bygade 19, DK-2860 Søborg, Denmark. anw@fdir.dk This study aimed to evaluate the possibility of DNA sequences from genetically modified plants to persist in the gastrointestinal (GI) tract. PCR analysis and transformation assays were used to study DNA persistence and integrity in various ex vivo and in vivo systems using gnotobiotic rats. DNA studied was either plasmid DNA, naked plant DNA or plant DNA embedded in maize flour. Ex vivo experiments performed by incubating plant DNA in intestinal samples, showed that DNA is rapidly degraded in the upper part of the GI tract whereas degradation is less severe in the lower part. In contrast, plasmid DNA could be recovered throughout the GI tract when intestinal samples were taken up to 5 h after feeding rats with plasmid. Furthermore, DNA isolated from these intestinal samples was able to transform electro-competent Escherichia coli, showing that the plasmid was still biologically active. The results indicate that ingested DNA may persist in the GI tract and consequently may be present for uptake by intestinal bacteria. Publication Types: Research Support, Non-U.S. Gov't PMID: 14871592 [PubMed - indexed for MEDLINE] 1335: Mol Ecol. 2004 Mar;13(3):639-47. Multitrophic interactions involving genetically modified potatoes, nontarget aphids, natural enemies and hyperparasitoids. Cowgill SE, Danks C, Atkinson HJ. Centre for Plant Sciences, School of Biology, University of Leeds, LS2 9JT, UK. S.E.Cowgill@leeds.ac.uk Genetically modified (GM) potatoes expressing a cysteine proteinase inhibitor (cystatin) have been developed as an option for the management of plant parasitic nematodes. The relative impact of such plants on predators and parasitoids (natural enemies) of nontarget insects was determined in a field trial. The trial consisted of GM plants, control plants grown in soil treated with a nematicide and untreated control plants. The quantity of nontarget aphids and their quality as hosts for natural enemies were studied. Aphid density was significantly reduced by nematicide treatment and few natural enemies were recorded from treated potatoes during the study. In contrast, similar numbers of aphids and their more abundant predators were recorded from the untreated control and the GM potatoes. The size of aphids on GM and control plants was recorded twice during the study. During the first sampling period (2-9 July) aphids clip-caged on GM plants were smaller than those on control plants. During the second sampling period (23-30 July) there was no difference in aphid size between those from the GM and control plants. Host size is an important component of host quality. It can affect the size and fecundity of parasitoid females and the sex ratio of their offspring. However, neither the fitness of females of Aphidius ervi, the most prevalent primary parasitoid, nor the sex ratio of their progeny, were affected when the parasitoids developed on aphids feeding on GM plants. Two guilds of secondary parasitoid were also recorded during the study. The fitness of the most abundant species, Aspahes vulgaris, was not affected when it developed on hosts from GM plants. The transgene product, OC I Delta D86, was not detected in aphids that had fed on GM plants in the field, suggesting that there is minimal secondary exposure of natural enemies to the inhibitor. The results indicate that transgenic nematode resistance is potentially more compatible with aphid biological control than is current nematicide use. Publication Types: Comparative Study Research Support, Non-U.S. Gov't PMID: 14871367 [PubMed - indexed for MEDLINE] 1336: Food Nutr Bull. 2003 Dec;24(4):303-18. Carotenoid-rich bananas: a potential food source for alleviating vitamin A deficiency. Englberger L, Darnton-Hill I, Coyne T, Fitzgerald MH, Marks GC. Nutrition Program, Division of International Health, School of Population Health, University of Queensland, Brisbane, Australia. nutrition@mail.fm This review article points out that bananas are an important food for many people in the world. Thus, banana cultivars rich in provitamin A carotenoids may offer a potential food source for alleviating vitamin A deficiency, particularly in developing countries. Many factors are associated with the presently known food sources of vitamin A that limit their effectiveness in improving vitamin A status. Acceptable carotenoid-rich banana cultivars have been identified in Micronesia, and some carotenoid-rich bananas have been identified elsewhere. Bananas are an ideal food for young children and families for many regions of the world, because of their sweetness, texture, portion size, familiarity, availability, convenience, versatility, and cost. Foods containing high levels of carotenoids have been shown to protect against chronic disease, including certain cancers, cardiovascular disease, and diabetes. Because the coloration of the edible flesh of the banana appears to be a good indicator of likely carotenoid content, it may be possible to develop a simple method for selecting carotenoid-rich banana cultivars in the community. Research is needed on the identification of carotenoid-rich cultivars, targeting those areas of the world where bananas are a major staple food; investigating factors affecting production, consumption, and acceptability; and determining the impact that carotenoid-rich bananas may have on improving vitamin A status. Based on these results, interventions should be undertaken for initiating or increasing homestead and commercial production. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 14870618 [PubMed - indexed for MEDLINE] 1337: Nature. 2004 Feb 5;427(6974):474. Europe urged to move on transgenic crop imports. Schiermeier Q. Publication Types: News PMID: 14765157 [PubMed - indexed for MEDLINE] 1338: Nat Biotechnol. 2004 Feb;22(2):170-2. Comment on: Nat Biotechnol. 2004 Feb;22(2):204-9. The fate of transgenes in the human gut. Heritage J. Publication Types: Comment News PMID: 14755289 [PubMed - indexed for MEDLINE] 1339: Nat Biotechnol. 2004 Feb;22(2):149. Comment on: Nat Biotechnol. 2003 Aug;21(8):835-6. Questions linger over European GM food regulations. Ramón D, MacCabe A, Gil JV. Publication Types: Comment Letter PMID: 14755281 [PubMed - indexed for MEDLINE] 1340: Nat Biotechnol. 2004 Feb;22(2):133. Erratum in: Nat Biotechnol. 2004 Apr;22(4):459. Comment in: Nat Biotechnol. 2004 Jul;22(7):803. Nat Biotechnol. 2004 Jul;22(7):803. Nat Biotechnol. 2004 Jun;22(6):655-6. Nat Biotechnol. 2004 May;22(5):507. Nat Biotechnol. 2004 May;22(5):508. Nat Biotechnol. 2004 May;22(5):508. Nat Biotechnol. 2006 Oct;24(10):1191-3. Drugs in crops--the unpalatable truth. [No authors listed] Publication Types: Editorial PMID: 14755274 [PubMed - indexed for MEDLINE] 1341: Curr Opin Allergy Clin Immunol. 2002 Dec;2(6):541-6. Animal models of food allergy. Helm RM, Burks AW. ACHRI/UAMS, Department of Pediatrics, Little Rock, Arkansas 72202, USA. helmrickim@uams.edu PURPOSE OF REVIEW: The focus of this review will be on recent animal models of food allergy. Animal models are being used to investigate underlying mechanisms of IgE-mediated disease and for prophylactic/intervention therapies to treat allergic disease. RECENT FINDINGS: Considerable advances have been made in the dosage and use of sensitization routes with and without adjuvant and determinations of the pathophysiology of food allergy in murine, dog and swine food allergy models. Continued research on the neuroendocrine and novel immunoregulatory peptides is also providing new insight into inflammatory regulation and immunity. With the advent of genetically modified food crops, animal models are becoming a central theme for prediction/assessment of allergenicity for novel proteins based upon known food allergens. Therapeutic strategies involving cytokine and allergen, DNA immunizations and the use of probiotics are receiving new interest. SUMMARY: Although murine models still predominate the literature with respect to animal models of food allergy, the atopic dog and neonatal swine model are contributing knowledge with respect to symptoms more closely related to human allergic responses. Continuing investigations into the mechanisms of IgE-mediated food allergy and therapeutic strategies are providing new insights into prevention and intervention therapies for food allergy. Publication Types: Comparative Study Review PMID: 14752339 [PubMed - indexed for MEDLINE] 1342: Plant Mol Biol. 2003 Oct;53(3):383-97. Molecular characterization of Brassica napus NAC domain transcriptional activators induced in response to biotic and abiotic stress. Hegedus D, Yu M, Baldwin D, Gruber M, Sharpe A, Parkin I, Whitwill S, Lydiate D. Molecular Genetics Section, Agriculture and Agri-Food Canada, 107 Science Place, Saskatoon, S7N OX2 Canada. hegedusd@em.agr.ca Subtractive expressed sequence tag analysis and screening of cDNA libraries derived from Brassica napus leaves subjected to mechanical wounding, flea beetle feeding or cold temperatures revealed eight genes encoding NAC-domain transcription factors. The genes were found to be differentially regulated in response to biotic and abiotic stresses including wounding, insect feeding, Sclerotinia sclerotiorum infection, cold shock and dehydration. Five BnNAC proteins were orthologous to Arabidopsis thaliana ATAF1 or ATAF2 and gave rise to developmental abnormalities similar to the A. thaliana nam and cuc mutants when expressed ectopically in A. thaliana. Transgenic lines expressing BnNAC14, exhibited large leaves, thickened stems and hyper-developed lateral root systems similar to that observed with A. thaliana NAC1, but also were delayed in bolting and lacked an apical dominant tap root. Several of the BnNAC proteins were capable of activating gene expression in yeast and recognized an element within the CaMV35S promoter. A yeast two-hybrid screen revealed that BnNAC14 interacted with other select BnNAC proteins in vitro and identified an additional BnNAC gene, BnNAC485. The protein interaction and transcriptional activation domains were mapped by deletion analysis. Publication Types: Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. PMID: 14750526 [PubMed - indexed for MEDLINE] 1343: J Nutr. 2004 Feb;134(2):431-4. Prima facie evidence that a phytocystatin for transgenic plant resistance to nematodes is not a toxic risk in the human diet. Atkinson HJ, Johnston KA, Robbins M. Centre for Plant Sciences, University of Leeds, Leeds, LS2 9JT, UK. h.j.atkinson@leeds.ac.uk A protein-engineered rice cystatin (OcIDeltaD86) provides transgenic, partial crop resistance to plant nematodes. This study determined whether its oral uptake has adverse effects on male Sprague-Dawley rats when they are administered by oral gavage 0.1-10 mg OcIDeltaD86/kg body weight daily for 28 d. Body weight and water and food intakes were unaltered for most of the study. The only significant changes in fresh weight of nine organs were for the liver (4% decrease; P < 0.05) and the empty cecum (14% increase; P < 0.05) at the two lowest doses and the highest dose of OcIDeltaD86, respectively. No abnormalities in either organ were detected by histochemistry. There were no changes in the urine or in hematological variables measured, and blood serum revealed no dose-dependent responses for any of 17 variables measured. OcIDeltaD86 was degraded by boiling with a 50% loss of its inhibition of papain after 9.2 +/- 8.0 min. It also showed >95% loss of such inhibition after 15 s in simulated gastric fluid. The results suggest that the no effect level (NOEL) for OcIDeltaD86 is >10 mg/(kg. d). This provides a range of dietary exposure >200-2000 fold depending upon the promoter used to control its expression in potato. Publication Types: Research Support, Non-U.S. Gov't PMID: 14747684 [PubMed - indexed for MEDLINE] 1344: Int Arch Allergy Immunol. 2004 Feb;133(2):101-12. Epub 2004 Jan 21. Statistical evaluation of local alignment features predicting allergenicity using supervised classification algorithms. Soeria-Atmadja D, Zorzet A, Gustafsson MG, Hammerling U. Division of Toxicology, National Food Administration, Uppsala University, Uppsala, Sweden. BACKGROUND: Recently, two promising alignment-based features predicting food allergenicity using the k nearest neighbor (kNN) classifier were reported. These features are the alignment score and alignment length of the best local alignment obtained in a database of known allergen sequences. METHODS: In the work reported here a much more comprehensive statistical evaluation of the potential of these features was performed, this time for the prediction of allergenicity in general. The evaluation consisted of the following four key components. (1) A new high quality database consisting of 318 carefully selected, non-redundant allergens and 1,007 sequences carefully selected to be non-allergens. (2) Three different supervised algorithms: the kNN classifier, the Bayesian linear Gaussian classifier, and the Bayesian quadratic Gaussian classifier. (3) A large set of local alignment procedures defined using the FASTA3 alignment program by means of a wide range of different parameter settings. (4) Novel performance curves, alternative to conventional receiver-operating characteristic curves, to display not only average behaviors but also statistical variations due to small data sets. RESULTS: The linear Gaussian classifier proved most useful among the tested supervised machine learning algorithms, closely followed by the quadratic Gaussian equivalent and kNN. The overall best classification results were obtained with a novel feature vector consisting of the combined alignment scores derived from local alignment procedures using different substitution matrices. CONCLUSIONS: The models reported here should be useful as a part of an integrated assessment scheme for potential protein allergenicity and for future comparisons with alternative bioinformatic approaches. Copyright 2004 S. Karger AG, Basel Publication Types: Evaluation Studies Research Support, Non-U.S. Gov't PMID: 14739578 [PubMed - indexed for MEDLINE] 1345: Science. 2004 Jan 23;303(5657):448-9. Genetically modified organisms. Europe takes tentative steps toward approval of commercial GM crops. Vogel G. Publication Types: News PMID: 14739427 [PubMed - indexed for MEDLINE] 1346: J Biotechnol. 2004 Feb 5;107(3):193-232. Selectable marker genes in transgenic plants: applications, alternatives and biosafety. Miki B, McHugh S. Research Branch, Agriculture and Agri-Food Canada, Room 2091, KW Neatby Bldg., CEF, 960 Carling Avenue, Ottawa, Ont., Canada K1A 0C6. mikib@agr.gc.ca Approximately fifty marker genes used for transgenic and transplastomic plant research or crop development have been assessed for efficiency, biosafety, scientific applications and commercialization. Selectable marker genes can be divided into several categories depending on whether they confer positive or negative selection and whether selection is conditional or non-conditional on the presence of external substrates. Positive selectable marker genes are defined as those that promote the growth of transformed tissue whereas negative selectable marker genes result in the death of the transformed tissue. The positive selectable marker genes that are conditional on the use of toxic agents, such as antibiotics, herbicides or drugs were the first to be developed and exploited. More recent developments include positive selectable marker genes that are conditional on non-toxic agents that may be substrates for growth or that induce growth and differentiation of the transformed tissues. Newer strategies include positive selectable marker genes which are not conditional on external substrates but which alter the physiological processes that govern plant development. A valuable companion to the selectable marker genes are the reporter genes, which do not provide a cell with a selective advantage, but which can be used to monitor transgenic events and manually separate transgenic material from non-transformed material. They fall into two categories depending on whether they are conditional or non-conditional on the presence of external substrates. Some reporter genes can be adapted to function as selectable marker genes through the development of novel substrates. Despite the large number of marker genes that exist for plants, only a few marker genes are used for most plant research and crop development. As the production of transgenic plants is labor intensive, expensive and difficult for most species, practical issues govern the choice of selectable marker genes that are used. Many of the genes have specific limitations or have not been sufficiently tested to merit their widespread use. For research, a variety of selection systems are essential as no single selectable marker gene was found to be sufficient for all circumstances. Although, no adverse biosafety effects have been reported for the marker genes that have been adopted for widespread use, biosafety concerns should help direct which markers will be chosen for future crop development. Common sense dictates that marker genes conferring resistance to significant therapeutic antibiotics should not be used. An area of research that is growing rapidly but is still in its infancy is the development of strategies for eliminating selectable marker genes to generate marker-free plants. Among the several technologies described, two have emerged with significant potential. The simplest is the co-transformation of genes of interest with selectable marker genes followed by the segregation of the separate genes through conventional genetics. The more complicated strategy is the use of site-specific recombinases, under the control of inducible promoters, to excise the marker genes and excision machinery from the transgenic plant after selection has been achieved. In this review each of the genes and processes will be examined to assess the alternatives that exist for producing transgenic plants. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 14736458 [PubMed - indexed for MEDLINE] 1347: Proteomics. 2004 Jan;4(1):193-200. Proteomics as a tool to improve investigation of substantial equivalence in genetically modified organisms: the case of a virus-resistant tomato. Corpillo D, Gardini G, Vaira AM, Basso M, Aime S, Accotto GP, Fasano M. Laboratorio Integrato Metodologie Avanzate, Bioindustry Park Canavese, Colleretto Giacosa, Italy. At present, the so-called "substantial equivalence" is the only widely accepted criterion for deciding whether or not a transgenic food is, from an alimentary point of view, to be considered totally correspondent to the "traditional" one from which it derives. Although never exactly defined, it deals with a comparison between the chemical composition of the two foods. A more in-depth analysis can be performed by one of the most suitable methods that allows for the simultaneous screening of many components without prior identification, the analysis of the proteome. As a model for testing this kind of approach, we compared protein expression of two types of tomato plants, having the same genetic background, except for a virus resistance trait introduced by genetic engineering. When proteins extracted from seedlings of the two types were analyzed by two-dimensional electrophoresis, no significant differences, either qualitative or quantitative, were detected, indicating that in this case the expression of major proteins was unmodified by the genetic manipulation. Fifteen proteins were identified by peptide mass fingerprinting. Publication Types: Research Support, Non-U.S. Gov't PMID: 14730681 [PubMed - indexed for MEDLINE] 1348: Nat Biotechnol. 2004 Feb;22(2):204-9. Epub 2004 Jan 18. Comment in: Nat Biotechnol. 2004 Feb;22(2):170-2. Nat Biotechnol. 2004 Jun;22(6):654-5. Assessing the survival of transgenic plant DNA in the human gastrointestinal tract. Netherwood T, Martín-Orúe SM, O'Donnell AG, Gockling S, Graham J, Mathers JC, Gilbert HJ. School of Cell and Molecular Biosciences, University of Newcastle upon Tyne, Newcastle upon Tyne NE1 7RU, UK. The inclusion of genetically modified (GM) plants in the human diet has raised concerns about the possible transfer of transgenes from GM plants to intestinal microflora and enterocytes. The persistence in the human gut of DNA from dietary GM plants is unknown. Here we study the survival of the transgene epsps from GM soya in the small intestine of human ileostomists (i.e., individuals in which the terminal ileum is resected and digesta are diverted from the body via a stoma to a colostomy bag). The amount of transgene that survived passage through the small bowel varied among individuals, with a maximum of 3.7% recovered at the stoma of one individual. The transgene did not survive passage through the intact gastrointestinal tract of human subjects fed GM soya. Three of seven ileostomists showed evidence of low-frequency gene transfer from GM soya to the microflora of the small bowel before their involvement in these experiments. As this low level of epsps in the intestinal microflora did not increase after consumption of the meal containing GM soya, we conclude that gene transfer did not occur during the feeding experiment. Publication Types: Clinical Trial Controlled Clinical Trial Research Support, Non-U.S. Gov't PMID: 14730317 [PubMed - indexed for MEDLINE] 1349: Plant Physiol. 2004 Jan;134(1):3-10. Production, identity preservation, and labeling in a marketplace with genetically modified and non-genetically modified foods. Huffman WE. Iowa State University, Ames, Iowa 50011, USA . Publication Types: Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. PMID: 14730058 [PubMed - indexed for MEDLINE] 1350: Johns Hopkins Med Lett Health After 50. 2001 Jul;13(5):8. Is genetically modified food safe? [No authors listed] PMID: 14727588 [PubMed - indexed for MEDLINE] 1351: Plant Cell Rep. 2004 Apr;22(9):684-90. Epub 2004 Jan 15. Pearl millet transformation system using the positive selectable marker gene phosphomannose isomerase. O'Kennedy MM, Burger JT, Botha FC. CSIR Food, Biological and Chemical Technologies, P.O. Box 395, 0001, Pretoria, South Africa. mokennedy@csir.co.za Fertile transgenic pearl millet plants expressing a phosphomannose isomerase (PMI) transgene under control of the maize ubiquitin constitutive promoter were obtained using the transformation system described here. Proliferating immature zygotic embryos were used as target tissue for bombardment using a particle inflow gun. Different culture and selection strategies were assessed in order to obtain an optimised mannose selection protocol. Stable integration of the manA gene into the genome of pearl millet was confirmed by PCR and Southern blot analysis. Stable integration of the manA transgene into the genome of pearl millet was demonstrated in T1 and T2 progeny of two independent transformation events with no more than four to ten copies of the transgene. Similar to results obtained from previous studies with maize and wheat, the manA gene was shown to be a superior selectable marker gene for improving transformation efficiencies when compared to antibiotic or herbicide selectable marker genes. PMID: 14727053 [PubMed - indexed for MEDLINE] 1352: Shi Yan Sheng Wu Xue Bao. 2003 Dec;36(6):428-34. [Double-antisense ACC oxidase and ACC synthase fusion gene introduced into tomato by Agrobacterium-mediated transformation and analysis the ethylene production of transgenic plants] [Article in Chinese] Xiong AS, Yao QH, Li X, Fan HQ, Peng RH. Shanghai Key Laboratory of Agricultural Genetics and Breeding, Agri-biotechnology Research Center, Shanghai Academy of Agricultural Science, Shanghai 201106, China. The tomato fruit-specific promoter 2A11 was amplified from tomato genomic DNA using PCR techniques. Total RNA was isolated from ripen fruit of tomato, then ACC oxidase gene and ACC synthase gene were obtained using reverse-transcription polymerase chain reaction. The fusion encoding ACC oxidase and ACC synthase gene was obtained through ACC oxidase gene and ACC synthase gene ligation. The fusion gene was then inserted into a plant binary vector pYPX145 in an inverted orientation. Finally, the binary plant expression vector pOSACC was constructed in which the double-antisense fusion gene was controlled by fruit-specific 2A11 promoter. By using hypocotyls and cotyledon petioles as explants, the unit of double-antisense fusion gene was successfully introduced into tomato (Lycopersicon esculentum Mill) cultivar "Hezuo 903" by Agrobacterium tumefaciens-mediated transformation. 105 transgenic plants were obtained through 200 mg/L kanamycin selection and GUS assay. Two lines of DR-1 and DR-2 were obtained through selecting the characteristics of prolonged shelf life and agriculture. The transgenic plants showed the characteristics of prolonged shelf life over 50 d. The amount of ethylene released from DR-1 and DR-2 fruits were reduced significantly to about 9.5% of that released by non-transformed controls. Publication Types: English Abstract Research Support, Non-U.S. Gov't PMID: 14724933 [PubMed - indexed for MEDLINE] 1353: Transgenic Res. 2003 Dec;12(6):671-81. Transgenic tobacco and apple plants expressing biotin-binding proteins are resistant to two cosmopolitan insect pests, potato tuber moth and lightbrown apple moth, respectively. Markwick NP, Docherty LC, Phung MM, Lester MT, Murray C, Yao JL, Mitra DS, Cohen D, Beuning LL, Kutty-Amma S, Christeller JT. The Horticulture and Food Research Institute of New Zealand Ltd, 120 Mt Albert Road, Auckland, New Zealand. nmarkwick@hortresearch.co.nz Tobacco (Nicotiana tabacum cv. Samsun) and apple (Malus x domestica cv. Royal Gala) plants expressing avidin or strepavidin were produced using Agrobacterium tumefaciens-mediated transformation. ELISA assays showed that avidin expression ranged from 3.1 to 4.6 microM in tobacco and from 1.9 to 11.2 microM in apple and streptavidin expression ranged from 11.4 to 24.5 microM in tobacco and from 0.4 to 14.6 microM in apple. Expressed at these levels, both biotin-binding proteins conferred a high level of insect resistance on transformed tobacco plants to larval potato tuber moth (PTM), Phthorimaea operculella (Zeller) (fam. Gelechiidae) and on apple plants to larvae of the lightbrown apple moth (LBAM) Epiphyas postvittana (Walker) (fam. Tortricidae). More than 90% of PTM larvae died on tobacco plants expressing either avidin or streptavidin genes within 9 days of inoculation. Mortality of LBAM larvae was significantly higher (P < 0.05) on three avidin-expressing (89.6, 84.9 and 80.1%) and two streptavidin-expressing (90 and 82.5%) apple plant lines than on non-transformed control plants (14.1%) after 21 days. Weight of LBAM larvae was also significantly reduced by feeding on all apple shoots expressing avidin and on apple shoots expressing streptavidin at levels of 3.8 microM and above. Publication Types: Research Support, Non-U.S. Gov't PMID: 14713196 [PubMed - indexed for MEDLINE] 1354: Transgenic Res. 2003 Dec;12(6):649-59. Concerted action of endogenous and heterologous phytase on phytic acid degradation in seed of transgenic wheat (Triticum aestivum L.). Brinch-Pedersen H, Hatzack F, Sørensen LD, Holm PB. Department of Plant Biology, Danish Institute of Agricultural Sciences, Research Centre Flakkebjerg, DK-4200 Slagelse, Denmark. henrik.brinchpedersen@agrsci.dk Expression of heterologous phytases in crops offers a great potential for improving phosphate and mineral bioavailability in food and feed. In this context it is of relevance to describe the concerted action of endogenous and hetrologous phytases on the transgenic seed inositol phosphate profile. Here we report metal-dye detection HPLC analysis of inositol phosphate degradation in flour from transgenic wheat materials possessing wheat endogenous 6-phytase [EC 3.1.3.26] and Aspergillus 3-phytase [EC 3.1.3.8] activities under the control of the maize ubiquitin-1 promoter and the wheat high molecular weight glutenin subunit 1DX5 promoter respectively. During 50 min incubation there is an accumulation of InsP5 to InsP2 breakdown products in non-transgenic material. Aspergillus niger phytase specific breakdown products are transiently detected in transgenic material but after 50 min incubation virtually all InsP5, InsP4 and InsP3 isomers are hydrolysed. PMID: 14713194 [PubMed - indexed for MEDLINE] 1355: Time. 2003 Dec 22;162(25):52. Got hormones? Roosevelt M. Publication Types: Legal Cases News PMID: 14712611 [PubMed - indexed for MEDLINE] 1356: J Cell Biol. 2004 Jan 5;164(1):47-56. Erratum in: J Cell Biol. 2004 Feb 16;164(4):following 623. Trafficking of plasmepsin II to the food vacuole of the malaria parasite Plasmodium falciparum. Klemba M, Beatty W, Gluzman I, Goldberg DE. Department of Molecular Microbiology, Washington University School of Medicine, 660 S. Euclid Ave., Box 8230, St. Louis, MO 63110, USA. A family of aspartic proteases, the plasmepsins (PMs), plays a key role in the degradation of hemoglobin in the Plasmodium falciparum food vacuole. To study the trafficking of proPM II, we have modified the chromosomal PM II gene in P. falciparum to encode a proPM II-GFP chimera. By taking advantage of green fluorescent protein fluorescence in live parasites, the ultrastructural resolution of immunoelectron microscopy, and inhibitors of trafficking and PM maturation, we have investigated the biosynthetic path leading to mature PM II in the food vacuole. Our data support a model whereby proPM II is transported through the secretory system to cytostomal vacuoles and then is carried along with its substrate hemoglobin to the food vacuole where it is proteolytically processed to mature PM II. Publication Types: Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. PMID: 14709539 [PubMed - indexed for MEDLINE] 1357: Int Arch Allergy Immunol. 2003 Dec;132(4):322-8. Development and use of an ELISA test to detect IgE antibody to Cry9c following possible exposure to bioengineered corn. Raybourne RB, Williams KM, Vogt R, Reissman DB, Winterton BS, Rubin C. Immunobiology Branch, Food and Drug Administration, Laurel, MD 20708, USA. richard.raybourne@cfsan.fda.gov BACKGROUND: Starlink(TM), a variety of corn genetically engineered to contain the insecticidal protein Cry9c, had not been approved for human consumption because it possessed some characteristics associated with allergenic proteins. However, in the fall of 2000 CRY9C DNA was detected in several corn-containing products, suggesting that Starlink corn had entered the human food supply. Subsequently, consumers, following consumption of corn products, reported a number of adverse health events, possibly consistent with allergic reaction. METHODS: To investigate the possibility of allergic reactions due to Cry9c in these consumers an ELISA test was developed for the purpose of detecting IgE antibodies to Cry9c and blood samples were taken from a total of 18 people who self-reported allergic reactions. Sera collected prior to the 1996 development of Starlink were used as negative controls. RESULTS: None of the adverse event sera were found to be reactive with recombinant Cry9c antigen, based on comparison with normal controls. Although a known human positive control serum containing IgE specific for Cry9c was not available, other controls were incorporated into the ELISA protocol, including the use of sera from subjects allergic to other allergens and their homologous antigens (cat, grass, peanut) to validate the IgE detection reagents. CONCLUSIONS: While the results do not support the likely occurrence of allergic reactions to Cry9c, such reactions cannot be ruled out, nor can the possibility that sera might react with unique glycosylated epitopes of Cry9c that may be expressed in the corn plant/seed. Copyright 2003 S. Karger AG, Basel PMID: 14707463 [PubMed - indexed for MEDLINE] 1358: Eur J Histochem. 2003;47(4):385-8. Fine structural analyses of pancreatic acinar cell nuclei from mice fed on genetically modified soybean. Malatesta M, Biggiogera M, Manuali E, Rocchi MB, Baldelli B, Gazzanelli G. We carried out ultrastructural morphometrical and immunocytochemical analyses on pancreatic acinar cell nuclei from mice fed on genetically modified (GM) soybean, in order to investigate possible structural and molecular modifications of nucleoplasmic and nucleolar constituents. We found a significant lowering of nucleoplasmic and nucleolar splicing factors as well as a perichromatin granule accumulation in GM-fed mice, suggestive of reduced post-transcriptional hnRNA processing and/or nuclear export. This is in accordance to already described zymogen synthesis and processing modifications in the same animals. Publication Types: Letter PMID: 14706936 [PubMed - indexed for MEDLINE] 1359: Nat Biotechnol. 2004 Jan;22(1):4-5. Vatican debates agbiotech. Fox JL. Publication Types: News PMID: 14704686 [PubMed - indexed for MEDLINE] 1360: Nat Biotechnol. 2004 Jan;22(1):1. Comment in: Nat Biotechnol. 2004 Apr;22(4):379; discussion 379. The one that got away. [No authors listed] Publication Types: Editorial PMID: 14704684 [PubMed - indexed for MEDLINE] 1361: Rocz Panstw Zakl Hig. 2004;55(4):279-86. [The investigation of presence of genetically modified protein in processed foodstuffs by ELISA test] [Article in Polish] Urbanek-Karłowska B, Sawilska-Rautenstrauch D, Jedra M. Zakład Badania Zywności i Przedmiotów Uzytku, Państwowy Zakład Higieny, 00-791 Warszawa, ul. Chocimska 24. The test based on immunoassay--TRAIT-RUR Toasted Soy Meal Kit was used for detection GMO-soy protein in the processed (heat treated) foodstuffs: bread, macaroni, sausages, ready-to-serve products and soya products (tofu, steaks). The threshold level is about 0,6% protein. The positive results were obtained for 27 from 106 investigated products. Only 5 foodstuffs were declared as containing GMO-soy in composition. The presence of genetically modified ingredients in foodstuffs must be controlled. The proper information should be labelled for the consumer. Publication Types: English Abstract PMID: 19097577 [PubMed - indexed for MEDLINE] 1362: Nutrition. 2004 Jan;20(1):121-6. Genetically modified mouse models for disorders of fatty acid metabolism: pursuing the nutrigenomics of insulin resistance and type 2 diabetes. Wood PA. Department of Genetics, University of Alabama at Birmingham, Birmingham, Alabama 35294-0024, USA. paw@uab.edu Publication Types: Research Support, U.S. Gov't, P.H.S. Review PMID: 14698026 [PubMed - indexed for MEDLINE] 1363: Trends Biotechnol. 2004 Jan;22(1):3. Greener vaccine production with GM yeasts. Wiseman A. Biochemistry Group, School of Biomedical and Life Sciences, University of Surrey, Guildford GU2 7XH, UK. PMID: 14690615 [PubMed - indexed for MEDLINE] 1364: J Biochem. 2003 Nov;134(5):765-72. Polyamine homeostasis in transgenic plants overexpressing ornithine decarboxylase includes ornithine limitation. Mayer MJ, Michael AJ. Institute of Food Research, Norwich Research Park, Colney, Norwich, NR4 7UA, United Kingdom. It was reported recently that overexpression of human ornithine decarboxylase (ODC) cDNA in transgenic rice plants resulted in increased steady-state concentration of polyamines, i.e., enough biosynthetic control is invested at this step to enable adjustment of polyamine levels. To investigate critically whether constitutive overexpression of ODC is sufficient to control steady-state polyamine levels, we expressed an ODC cDNA from Datura stramonium in transgenic tobacco plants. Transgenic progeny of self-fertilised primary transformants exhibited increases in ODC activity of 25-fold in leaves and 5-fold in flower buds. However, the increase in putrescine levels was only 1.5- to 2.1-fold in leaves and 1.1- to 1.3-fold in flower buds. Emphatically, no changes to spermidine or spermine steady-state levels or to soluble or insoluble hydroxycinnamic acid-conjugated polyamines were observed. Ornithine feeding to cell suspension cultures derived from the transgenic plants indicated that putrescine accumulation was limited in part by ornithine availability. These results demonstrate that a large increase in the capacity of the tobacco plants to decarboxylate ornithine does not result in a comparable increase in the level of free or conjugated polyamines. Plant polyamine homeostatic mechanisms efficiently accommodate increased ODC activity, suggesting that polyamine biosynthetic control is invested at multiple interdependent steps. PMID: 14688243 [PubMed - indexed for MEDLINE] 1365: J Anim Sci. 2003 Dec;81(12):3216-7. Symposium on analytical method challenges for measuring nutrients and antinutrients in plants: introduction to the symposium. Price WD. Office of Surveillance and Compliance, Center for Veterinary Medicine, Food and Drug Administration, Rockville, MD 20855, USA. wprice@cvm.fda.gov Publication Types: Congresses PMID: 14677878 [PubMed - indexed for MEDLINE] 1366: Nature. 2003 Dec 11;426(6967):591-5914. Berkeley accused of biotech bias as ecologist is denied tenure. Dalton R. Publication Types: News PMID: 14668822 [PubMed - indexed for MEDLINE] 1367: Syst Appl Microbiol. 2003 Nov;26(4):495-501. Effect of food processing on the fate of DNA with regard to degradation and transformation capability in Bacillus subtilis. Kharazmi M, Bauer T, Hammes WP, Hertel C. Institute of Food Technology, University of Hohenheim, Stuttgart, Germany. Soymilk, tofu, corn masa, and cooked potato were produced from transgenic raw materials and the effect of processing on the degradation of DNA was studied. Major degrading factors were for soymilk and tofu the mechanical treatment of soaked soybeans and for corn masa and cooked potatoes the thermal treatment. In the processed foods no DNA fragments > 1.1 kb were detected. We included in our studies the effect of the size of donor DNA and length of the homologous sequence on the marker rescue transformation of B. subtilis LTH 5466, which was monitored by restoration of deleted nptII. When DNA fragments (168, 414, 658, and 792 bp) of nptII and linearized plasmid DNA (pGEM-T-1, 3168 bp and pGEM-T-2, 3792 bp) containing the 168 bp or 792 bp fragments, respectively, were used as donor DNA, it was observed that the efficiency of marker rescue decreased with decreasing length of homologous sequence. The use of a larger plasmid (pMR2, 5786 bp) containing the 792 bp fragment revealed higher efficiency of marker rescue compared to pGEM-T-2. The nptII fragments resulted in lower efficiencies compared to plasmid DNA containing the same fragment. For the 792 bp fragment and the linearized plasmid pMR2 a first-order dependency of the frequency of marker rescue transformation on the DNA concentration was observed. Based on the acquired data, the hypothetical frequency of transformation of transgenic DNA to B. subtilis in cooked potatoes was calculated to be equal to 8.5 x 10(-19) and 1.2 x 10(-27) for homologous and illegitimate recombination, respectively. These data permit to roughly estimate the time after which a person (10(8) years) or the world population (15 days) is exposed to one transformant generated by homologous recombination event, when the daily consumption per person is 130 g of cooked potatoes. Publication Types: Research Support, Non-U.S. Gov't PMID: 14666975 [PubMed - indexed for MEDLINE] 1368: Trends Plant Sci. 2003 Dec;8(12):591-7. Tracking genes from seed to supermarket: techniques and trends. Auer CA. Department of Plant Science, University of Connecticut, Storrs, CT 06269-4163, USA. carol.auer@uconn.edu Analytical techniques to track plant genes in the environment and the food chain are essential for environmental risk assessment, government regulation and production and trade of genetically modified (GM) crops. Here, I review laboratory techniques to track plant genes during pre-commercialization research on gene flow and post-commercialization detection, identification and quantification of GM crops from seed to supermarket. At present, DNA- and protein-based assays support both activities but the demand for fast, inexpensive, sensitive methods is increasing. Part of the demand has been generated by stringent food labeling and traceability regulations for GM crops. The increase in GM crops, changes in GM crop design, evolution of government regulations and adoption of risk-assessment frameworks will continue to drive development of analytical techniques. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 14659708 [PubMed - indexed for MEDLINE] 1369: Nature. 2003 Dec 4;426(6966):495. Comment on: Nature. 2003 Oct 9;425(6958):559. GM-debate methodology works in the real world. Grove-White R. Publication Types: Comment Letter PMID: 14654814 [PubMed - indexed for MEDLINE] 1370: Curr Biol. 2003 Dec 2;13(23):R889-90. Researchers protest at GM crop debate. Williams N. Publication Types: News PMID: 14654005 [PubMed - indexed for MEDLINE] 1371: Nature. 2003 Nov 27;426(6965):372. GloFish casts light on murky policing of transgenic animals. Knight J. Publication Types: News PMID: 14647345 [PubMed - indexed for MEDLINE] 1372: Nat Biotechnol. 2003 Dec;21(12):1429-30. UK field-scale evaluations answer wrong questions. Chassy B, Carter C, McGloughlin M, McHughen A, Parrott W, Preston C, Roush R, Shelton A, Strauss SH. Publication Types: Letter PMID: 14647318 [PubMed - indexed for MEDLINE] 1373: Mutat Res. 2003 Nov;544(2-3):223-33. Genetically modified crops: environmental and human health concerns. Azevedo JL, Araujo WL. Departamento de Genética, Escola Superior de Agricultura, Luiz de Queiroz, Universidade de São Paulo, P.O. Box 83, 13400-970 Piracicaba, São Paulo, Brazil. jazevedo@esalq.usp.br About 10,000 years ago subsistence farmers started to domesticate plants and it was only much later, after the discovery of the fundaments of genetics, those organisms were submitted to rational genetic improvement mainly by selecting of traits of interest. Breeders used appropriate gene combinations to produce new animal races, plant varieties and hybrids, as well as improved microorganisms such as yeasts. After the introduction of recombinant DNA techniques, the transfer of DNA between species belonging to different genera, families or kingdoms became possible. The release of transgenic plants has aroused debates about several aspects of the environmental and human risks that could result from the introduction of genetically modified crops. Less effort has been dedicated to evaluate the impact of transgenic plants on their associated microorganisms, some of which (e.g. nitrogen-fixing bacteria, mycorrhizal fungi and endophytic microbiota) are extremely important for the survival of the plant. Investigations have been made regarding the horizontal transfer of genetic material between transgenic plants and microorganisms and on the disturbance of useful symbiotic associations between plants and endophytic, epiphytic and rhizosphere communities. In most cases the results do no show any adverse effect of transgenic plants on autochthonous plant-associated microorganisms. Results from our laboratory show small changes caused by genetically modified endophytic bacteria on the indigenous endophytic population of the sweet orange Citrus sinensis. In tests using appropriated fungal strains preliminary results using extracts from transgenic plants indicate that these plants do not affect haploidization, mitotic crossing-over, mutation rate or chromosomal alterations. Publication Types: Review PMID: 14644324 [PubMed - indexed for MEDLINE] 1374: Mutat Res. 2003 Nov;544(2-3):217-21. Human health concerns with GM crops. Malarkey T. Syngenta Biotechnology Inc., P.O. Box 12257, 3054 Cornwallis Road, Research Triangle Park, NC 27709, USA. trish.malarkey@syngenta.com Biotechnology was used in the first generation of so-called 'GM' crops to provide growers with complimentary and sometimes alternative crop management solutions to pesticides. Selected host genes or genes identified from other plants or non-plant sources are modified or transferred to a crop plant. The new or altered protein expression resulting from these modifications confer on the plant a desired physiological trait, such as resistance to particular herbicides or insect pests. Second generation modifications provide traits such as enhanced nutritional or health-promoting characteristics that are of benefit to consumers. The commonly raised concerns about possible implications for human health are: inherent toxicity of the novel gene and their products, the potential to express novel antigenic proteins or alter levels of existing protein allergens, the potential for unintended effects resulting from alterations of host metabolic pathways or over expression of inherently toxic or pharmacologically active substances and the potential for nutrient composition in the new food occur differing significantly from a conventional counterpart. Foods produced using biotechnology are subjected to far greater levels of scrutiny than foods produced by traditional plant breeding techniques. The accepted analytical, nutritional and toxicological methods employed to support this scrutiny and to assess and assure that a 'GM' food is a safe and nutritious as its 'non-GM' counterpart are discussed. The challenges associated with identifying unintended effects in whole GM foods and the promise new (proteomics/genomic) technologies offer opposite traditional toxicity testing paradigms are appraised. Publication Types: Review PMID: 14644323 [PubMed - indexed for MEDLINE] 1375: Metab Eng. 2003 Oct;5(4):255-63. Coordinate expression of multiple bacterial carotenoid genes in canola leading to altered carotenoid production. Ravanello MP, Ke D, Alvarez J, Huang B, Shewmaker CK. Monsanto Company, Calgene Campus, 1920 Fifth St, Davis, CA 95616, USA. monica.p.ravello@monsanto.com Carotenoids have drawn much attention recently because of their potentially positive benefits to human health as well as their utility in both food and animal feed. Previous work in canola (Brassica napus) seed over-expressing the bacterial phytoene synthase gene (crtB) demonstrated a change in carotenoid content, such that the total levels of carotenoids, including phytoene and downstream metabolites like beta-carotene, were elevated 50-fold, with the ratio of beta- to alpha-carotene being 2:1. This result raised the possibility that the composition of metabolites in this pathway could be modified further in conjunction with the increased flux obtained with crtB. Here we report on the expression of additional bacterial genes for the enzymes geranylgeranyl diphosphate synthase (crtE), phytoene desaturase (crtI) and lycopene cyclase (crtY and the plant B. napus lycopene beta-cyclase) engineered in conjunction with phytoene synthase (crtB) in transgenic canola seed. Analysis of the carotenoid levels by HPLC revealed a 90% decrease in phytoene levels for the double construct expressing crtB in conjunction with crtI. The transgenic seed from all the double constructs, including the one expressing the bacterial crtB and the plant lycopene beta-cyclase showed an increase in the levels of total carotenoid similar to that previously observed by expressing crtB alone but minimal effects were observed with respect to the ratio of beta- to alpha-carotene compared to the original construct. However, the beta- to alpha-carotene ratio was increased from 2:1 to 3:1 when a triple construct consisting of the bacterial phytoene synthase, phytoene desaturase and lycopene cyclase genes were expressed together. This result suggests that the bacterial genes may form an aggregate complex that allows in vivo activity of all three proteins through substrate channeling. This finding should allow further manipulation of the carotenoid biosynthetic pathway for downstream products with enhanced agronomic, animal feed and human nutritional values. PMID: 14642353 [PubMed - indexed for MEDLINE] 1376: J Nutr Educ Behav. 2003 Nov-Dec;35(6):319-32. Application of modern biotechnology to food and agriculture: food systems perspective. McCullum C, Benbrook C, Knowles L, Roberts S, Schryver T. Center for Health Promotion and Prevention Research, School of Public Health, University of Texas-Houston, Health Science Center, Houston, Texas 77030, USA. christine.mccullum@uth.tmc.edu The purpose of this article is to provide nutrition educators with an introduction to a range of considerations and forces that are driving the application of modern biotechnology in the food and fiber sector based on a food systems perspective. In doing so, the following issues are critically assessed: (1) the global debate on how to regulate genetically engineered (GE) foods and crops, (2) cultural differences in public perceptions of GE foods, and (3) evaluation of selected GE traits against the principles of social, economic, and ecological sustainability, including the potential of modern agricultural biotechnology to enhance global food security. Where appropriate, we also review other agricultural technologies and the broader political, social, and economic contexts in which these technologies have been introduced. Finally, we offer recommendations for how multiple stakeholder groups, including policy makers, biotechnology advocates, and nutrition educators, can move toward a more informed dialogue and debate on this issue. Publication Types: Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. Review PMID: 14642218 [PubMed - indexed for MEDLINE] 1377: Risk Anal. 2003 Dec;23(6):1117-33. Communicating about the risks and benefits of genetically modified foods: the mediating role of trust. Frewer LJ, Scholderer J, Bredahl L. Wageningen University and Research Centres, Department of Marketing and Consumer Behavior, Wageningen, The Netherland. Lynn.Frewer@wur.nl Recent research suggests that public attitudes toward emerging technologies are mainly driven by trust in the institutions promoting and regulating these technologies. Alternative views maintain that trust should be seen as a consequence rather than a cause of such attitudes. To test its actual role, direct as well as mediating effects of trust were tested in an attitude change experiment involving 1,405 consumers from Denmark, Germany, Italy, and the United Kingdom. After prior attitudes to genetic modification in food production had been assessed, participants received different information materials (either product-specific information or balanced/general information about genetic modification in food production) and were asked to evaluate different types of genetically modified foods (either beer or yoghurt). The information materials were attributed to different information sources (either an industry association, a consumer organization, or a government source). After completion, perceived risk and perceived benefit were assessed, and participants indicated their trust in the information sources to which the materials had been attributed. Direct and trust-mediated attitude change effects were estimated in a multi-sample structural equation model. The results showed that information provision had little effect on people's attitudes toward genetically modified foods, and that perceptions of information source characteristics contributed very little to attitude change. Furthermore, the type of information strategy adopted had almost no impact on postexperimental attitudes. The extent to which people trusted the information sources appeared to be driven by people's attitudes to genetically modified foods, rather than trust influencing the way that people reacted to the information. Trust was not driving risk perception-rather, attitudes were informing perceptions of the motivation of the source providing the information. Publication Types: Research Support, Non-U.S. Gov't PMID: 14641888 [PubMed - indexed for MEDLINE] 1378: J Agric Food Chem. 2003 Dec 3;51(25):7450-5. Simultaneous transgenic suppression of LePG and LeExp1 influences fruit texture and juice viscosity in a fresh market tomato variety. Powell AL, Kalamaki MS, Kurien PA, Gurrieri S, Bennett AB. Department of Vegetable Crops, University of California, Davis, California 95616. alpowell@ucdavis.edu Tomatoes are grown for fresh consumption or for processing of the fruit. Some ripening-associated processes of the fruit can either contribute to or degrade attributes associated with both fresh and processing quality. For example, cell wall disassembly is associated with loss of fresh fruit firmness as well as with loss of processed tomato product viscosity. Several enzymes contribute to cell wall polysaccharide disassembly. Polygalacturonase (PG, poly[1,4-alpha-d-galactouronide] glucanohydrolase, EC 3.2.1.15) is among the most abundant polysaccharide hydrolases in ripening tomato fruit and is the major contributor to pectin depolymerization. Expansin (LeExp1) is also abundant in ripening fruit and is proposed to contribute to cell wall disassembly by nonhydrolytic activity, possibly by increasing substrate accessibility to other enzymes. Suppression of either LePG or LeExp1 expression alone results in altered softening and/or shelf life characteristics. To test whether simultaneous suppression of both LePG and LeExp1 expression influences fruit texture in additive or synergistic ways, transgenic Lycopersicon esculentum var. Ailsa Craig lines with reduced expression of either LePG or LeExp1 were crossed. Fruits from the third generation of progeny, homozygous for both transgenic constructs, were analyzed for firmness and other quality traits during ripening on or off the vine. In field-grown transgenic tomato fruit, suppression of LeExp1 or LePG alone did not significantly increase fruit firmness. However, fruits suppressed for both LePG and LeExp1 expression were significantly firmer throughout ripening and were less susceptible to deterioration during long-term storage. Juice prepared from the transgenic tomato fruit with reduced LePG and LeExp1 expression was more viscous than juice prepared from control fruit. PMID: 14640598 [PubMed - indexed for MEDLINE] 1379: J Agric Food Chem. 2003 Dec 3;51(25):7465-71. Transgenic overexpression of expansin influences particle size distribution and improves viscosity of tomato juice and paste. Kalamaki MS, Powell AL, Struijs K, Labavitch JM, Reid DS, Bennett AB. Department of Food Science, Vegetable Crops, University of California, Davis, California 95616. Suppression of the expression of a ripening-related expansin gene, LeExp1, in tomato enhanced fruit firmness and overexpression of LeExp1 resulted in increased fruit softening. Because of the incompletely understood relationship between fresh fruit texture and the consistency of processed products, we examined the effects of LeExp1 overexpression on the processing characteristics of tomato fruit. As determined by Bostwick consistency and by controlled strain rheometry, juices and pastes prepared from transgenic tomatoes with suppressed LeExp1 expression had a higher viscosity than preparations from control fruits. However, the viscosity of juice and paste prepared from fruit overexpressing LeExp1 was significantly greater than products from controls or lines with reduced LeExp1. Bostwick consistency increased by 9% (juice) and 6% (paste) in lines with suppressed LeExp1 expression but increased by 27.5% (juice) and 19.5% (paste) in lines overexpressing LeExp1, relative to controls. Determined by laser diffraction, the particles in juice and paste prepared from transgenic fruits with reduced LeExp1 expression were smaller, and preparations from fruits overexpressing LeExp1 had a size distribution indicating more large particles. Analysis of cell wall polysaccharides size indicated that LeExp1 overexpression enhanced depolymerization of water soluble pectins as well as tightly bound matrix glycans. LeExp1 overexpression may allow increased cell wall hydration, resulting in expanded particle size and increased viscosity of products. Because either LeExp1 suppression or overexpression leads to improved consistency, the interactions that contribute to optimal product rheological properties are complex. Publication Types: Research Support, Non-U.S. Gov't PMID: 14640600 [PubMed - indexed for MEDLINE] 1380: J Agric Food Chem. 2003 Dec 3;51(25):7456-64. Simultaneous transgenic suppression of LePG and LeExp1 influences rheological properties of juice and concentrates from a processing tomato variety. Kalamaki MS, Harpster MH, Palys JM, Labavitch JM, Reid DS, Brummell DA. Department of Food Science and Technology, University of California, Davis, California 95616. Processing tomato lines suppressed in the accumulation of ripening-related polygalacturonase or expansin were generated by introduction of transgenes to silence expression of the LePG and LeExp1 genes, respectively. The rheological properties of juice and juice reconstituted from paste produced from lines suppressed in one of these genes, or in both, were compared with azygous controls. When assayed by measuring Bostwick consistency, paste produced from either suppressed LePG or suppressed LeExp1 lines and diluted to 5 degrees Brix was approximately 18% more viscous than that produced from controls. Simultaneous suppression of LePG and LeExp1 produced a small additional increase in viscosity of 4%. Rheometric flow analysis at 5 or 10 degrees Brix also showed substantial increases in the consistency index due to suppression of either LePG or LeExp1 alone, and a small additional increase when both genes were suppressed in the same transgenic line. Measurements by laser diffraction and [1H]NMR showed that suppression of LePG or LeExp1 accumulation altered the size distribution of insoluble particles and modified their surface properties. The data are consistent with suppression of LePG increasing serum viscosity, and suppression of either LePG or LeExp1 altering the properties of the insoluble particles and improving some aspect of particle-particle or particle-serum interaction, or both. However, relative to that caused by suppression of either gene alone, the additional increase in viscosity caused by simultaneous suppression of LePG and LeExp1 together was slight. Publication Types: Research Support, Non-U.S. Gov't PMID: 14640599 [PubMed - indexed for MEDLINE] 1381: Anal Chem. 2003 Sep 1;75(17):392A-396A. Tailoring thresholds for GMO testing. Ruth L. PMID: 14632033 [PubMed - indexed for MEDLINE] 1382: Plant Cell. 2003 Dec;15(12):3007-19. Epub 2003 Nov 20. Engineering vitamin E content: from Arabidopsis mutant to soy oil. Van Eenennaam AL, Lincoln K, Durrett TP, Valentin HE, Shewmaker CK, Thorne GM, Jiang J, Baszis SR, Levering CK, Aasen ED, Hao M, Stein JC, Norris SR, Last RL. Monsanto Company, Calgene Campus, Davis, California 95616, USA. We report the identification and biotechnological utility of a plant gene encoding the tocopherol (vitamin E) biosynthetic enzyme 2-methyl-6-phytylbenzoquinol methyltransferase. This gene was identified by map-based cloning of the Arabidopsis mutation vitamin E pathway gene3-1 (vte3-1), which causes increased accumulation of delta-tocopherol and decreased gamma-tocopherol in the seed. Enzyme assays of recombinant protein supported the hypothesis that At-VTE3 encodes a 2-methyl-6-phytylbenzoquinol methyltransferase. Seed-specific expression of At-VTE3 in transgenic soybean reduced seed delta-tocopherol from 20 to 2%. These results confirm that At-VTE3 protein catalyzes the methylation of 2-methyl-6-phytylbenzoquinol in planta and show the utility of this gene in altering soybean tocopherol composition. When At-VTE3 was coexpressed with At-VTE4 (gamma-tocopherol methyltransferase) in soybean, the seed accumulated to >95% alpha-tocopherol, a dramatic change from the normal 10%, resulting in a greater than eightfold increase of alpha-tocopherol and an up to fivefold increase in seed vitamin E activity. These findings demonstrate the utility of a gene identified in Arabidopsis to alter the tocopherol composition of commercial seed oils, a result with both nutritional and food quality implications. PMID: 14630966 [PubMed - indexed for MEDLINE] 1383: Nature. 2003 Nov 20;426(6964):224-6. GM crops: a continent divided. Masood E. Publication Types: News PMID: 14628018 [PubMed - indexed for MEDLINE] 1384: J Insect Physiol. 2003 Dec;49(12):1199-209. Probiotic effects of beta-glucuronidase on the peach-potato aphid Myzus persicae (Aphididae). Cherqui A, Alla S, Saguez J, Doury G, Sangwan-Norreel BS, Giordanengo P. Laboratoire de Biologie des Entomophages, UPRES EA 2084, Université de Picardie Jules Verne, 33 rue Saint Leu, 80039 Amiens Cedex, France. beta-glucuronidase (GUS) is a reporter protein commonly expressed in transgenic plants allowing the visualization of the transformed individuals. In our recent work, we showed that consumption of transformed potato plants expressing this GUS enzyme improves performance of the phloem feeding aphid Myzus persicae. Those results led us to the conclusion that the expression of GUS in potato plants might be responsible for the probiotic effect measured in feeding aphids. In the present paper, artificial diets were used to provide active GUS (10 and 500 microg ml(-1)), inactivated heated GUS (500 microg ml(-1)), glucuronic acid (10, 100 and 500 microg ml(-1)), and bovine serum albumin (500 microg ml(-1)) to M. persicae. Our results reveal that these chemicals provided as food intake might influence the biological parameters of this aphid. Experiments showed a probiotic effect of 500 microg ml(-1) GUS diet, resulting in reduced larval mortality, and increased adult reproduction period and fecundity, which led to an increased population growth potential (r(m)=0.17+/-0.01 versus r(m)=0.12+/-0.03 for aphids fed on control diet). A lower amount of added GUS led to fewer variations, biological parameters being only slightly altered (r(m)=0.14+/-0.03). Statistically similar alterations of the biological parameters were obtained when comparing aphids fed on the diet added with inactivated GUS or the non-structural bovine serum albumin protein (r(m)=0.15+/-0.02 and 0.14+/-0.03, respectively). Feeding assays conducted with glucuronic acid supplemented diets enhanced longevity and nymph production of the adult aphids and reduced larval mortality, resulting in r(m)=0.15+/-0.02 for the highest dose (500 microg ml(-1)). Although 100 microg ml(-1) glucuronate diet did not induce any effect on M. persicae (r(m)=0.12+/-0.03), aphids fed on 10 microg ml(-1) glucuronate diet exhibited unexpected reduced demographic parameters (r(m)=0.10+/-0.03). Immuno-histological analysis showed GUS labeling along the whole digestive epithelium of adults and in various tissues including embryos and bacteriocytes. These results suggest that GUS crosses through the digestive tract. Western blots performed with protein extracts of transformed potato plants expressing the gus gene showed a unique band of molecular weight 76 kDa. On the contrary, in extracts from aphids fed on transgenic potato plants or bred on GUS 500 microg ml(-1) artificial diet, several proteins of lower molecular weight were hybridized, revealing proteolysis of ingested GUS. It is concluded that GUS protein, and more precisely GUS activity, is responsible for the probiotic effects on aphid feeding. The possible pathways of induction of such physiological alterations by GUS are discussed. Publication Types: Research Support, Non-U.S. Gov't PMID: 14624892 [PubMed - indexed for MEDLINE] 1385: Biotechnol Adv. 2003 Dec;22(1-2):45-69. Sustainability of insect resistance management strategies for transgenic Bt corn. Glaser JA, Matten SR. United States Environmental Protection Agency, Office of Research & Development, National Risk Management Research Laboratory, Sustainable Technology Division, 26 W King Dr., Cincinnati, OH 45268, USA. glaser.john@epa.gov Increasing interest in the responsible management of technology in the industrial and agricultural sectors of the economy has been met thorough the development of broadly applicable tools to assess the "sustainability" of new technologies. An arena ripe for application of such analysis is the deployment of transgenic crops. The new transgenic pesticidal or plant-incorporated protectant (PIP) crops have seen widespread application in the United States based on the features of higher yield, lower applications of insecticides, and control of mycotoxin content. However, open rejection of these new crops in Europe and in other countries has been a surprising message and has limited their worldwide acceptance. The US Environmental Protection Agency's (USEPA) Office of Pesticide Programs (OPP) has worked on the development and analysis of insect resistance management (IRM) strategies and has mandated specific IRM requirements for Bacillus thuringiensis (Bt) crops since 1995 under the Food, Fungicide, Insecticide, and Rodenticide Act. Improvement of data quality and sustainability of IRM strategies have been targeted in an ongoing partnership between the USEPA Office of Research and Development and the Office of Pesticide Programs that will further enhance the agency's ability to develop sustainable insect resistance management strategies for transgenic field corn (Bt corn) producing B. thuringiensis (Bt) insecticidal proteins. PMID: 14623043 [PubMed - indexed for MEDLINE] 1386: Public Underst Sci. 2002 Apr;11(2):93-111. Controversial medical and agri-food biotechnology: a cultivation analysis. Bauer MW. Social Psychology, London School of Economics, Department of Social Psychology, UK. M.Bauer@lse.ac.uk Whether biotechnology is one or several developments is not clear. Once distinctions are required, the question is: Which one prevails? When the good, the bad, and the ugly settle, where do they fall? Evaluation implies distinction, and representation drives attitude. The controversies over biotechnology are fertile ground on which to study these issues. The imports of genetically modified (GM) soya into Europe in 1996-97 and the cloning of Dolly the sheep from adult cells in 1997 changed the symbolic environment for genetic engineering. The ensuing public controversies came to focus mainly on field trials of GM crops and food labeling. This paper will explore the relationship between quality press coverage and public perception, in particular the cultivation of the contrast between "desirable" biomedical (RED) and "undesirable" agri-food (GREEN) biotechnology in Britain. The argument draws on a systematic analysis of the British press coverage of biotechnology from 1973 to 1999 and analysis of public perceptions in 1996 and 1999. The paper concludes that the debate over GM crops and food ingredients fostered the RED-GREEN contrast among the newspaper-reading public, thereby shielding RED biotechnology from public controversy, and ushered in a realignment of the regulatory framework in 2000. Publication Types: Historical Article PMID: 14621673 [PubMed - indexed for MEDLINE] 1387: Camb Q Healthc Ethics. 2003 Fall;12(4):342-52. Biotechnology and the fear of Frankenstein. Campbell CS. Program for Ethics, Science, and the Environment, Department of Philosophy, Oregon State University, Corvallis, Oregon, USA. PMID: 14619367 [PubMed - indexed for MEDLINE] 1388: Proc Natl Acad Sci U S A. 2003 Nov 25;100(24):14217-22. Epub 2003 Nov 14. Sustained peripheral expression of transgene adiponectin offsets the development of diet-induced obesity in rats. Shklyaev S, Aslanidi G, Tennant M, Prima V, Kohlbrenner E, Kroutov V, Campbell-Thompson M, Crawford J, Shek EW, Scarpace PJ, Zolotukhin S. Department of Molecular Genetics and Microbiology, Powell Gene Therapy Center, University of Florida, Gainesville, FL 32610-0266, USA. Adiponectin (Acrp30) is a physiologically active polypeptide hormone secreted by adipose tissue that shows insulin-sensitizing, antiinflammatory, and antiatherogenic properties. In humans, Acrp30 levels are inversely related to the degree of adiposity. In the current study, we tested the long-term weight-reducing and insulin-enhancing effects of Acrp30 cDNA delivered peripherally by a viral vector. To this end, we have generated a series of recombinant adeno-associated virus vectors of serotypes 1 and 5 encoding mouse Acrp30 cDNAs. The long-term expression of recombinant adeno-associated virus-Acrp30 vectors was tested after intramuscular or intraportal injection in female Sprague-Dawley rats with diet-induced obesity. We show that a single peripheral injection of 10(12) physical particles of Acrp30-encoding vectors resulted in sustained (up to 280 days) significant reduction in body weight, concomitant with the reduction in daily food intake. Acrp30 treatment resulted in higher peripheral insulin sensitivity measured by the i.p. glucose tolerance test in fasted animals. Ectopic expression of the Acrp30 transgene resulted in modulation of hepatic gluconeogenesis and lipogenesis, as demonstrated by the reduction of the expression of two key genes: PEPCK (phosphoenolpyruvate carboxykinase) and SREBP-1c (sterol regulatory element-binding protein 1c) in the liver. These data show successful peripheral therapy in a clinically relevant model for human obesity and insulin resistance. Publication Types: Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S. PMID: 14617771 [PubMed - indexed for MEDLINE] 1389: J Allergy Clin Immunol. 2003 Nov;112(5):1011-2. Comment in: J Allergy Clin Immunol. 2004 May;113(5):1003-4; author reply 1004-5. A negative, double-blind, placebo-controlled challenge to genetically modified corn. Sutton SA, Assa'ad AH, Steinmetz C, Rothenberg ME. Publication Types: Case Reports Letter Research Support, Non-U.S. Gov't PMID: 14610498 [PubMed - indexed for MEDLINE] 1390: J Nutr. 2003 Nov;133(11 Suppl 1):3830S-3836S. Diet, nutrition, and cancer prevention: the postgenomic era. Go VL, Butrum RR, Wong DA. UCLA Center for Human Nutrition, David Geffen School of Medicine at UCLA, Los Angeles, CA 90095, USA. The genomic era of human nutrition is upon us: the human genome and several plant genomes have been characterized, and genetically modified foods are now abundantly available in the marketplace. The link between diet and cancer is well established, and new genomic technologies have made possible the investigation of nutritional modulation of the carcinogenesis pathway with nutrients, micronutrients, and phytochemicals. Current study of nutrient-modulated carcinogenesis involves exploring the effect of nutrients on DNA damage and repair mechanisms; DNA methylation, which influences gene expression and cellular phenotypes; antioxidant rearranging and oxidative stress; target receptors and signal transduction pathways; cell cycle controls and check points; apoptosis; and antiangiogenic processes. With nutritional genomics, proteomics, and metabolomics, scientists are able to simultaneously elucidate the biological effects of dietary constituents on cell function and global gene expression. This generation of new knowledge on nutrient-gene interactions provides the justification for a research framework for diet and cancer prevention that is focused on identifying and developing new biomarkers as well as a novel and contemporary paradigm for dietary intervention. Publication Types: Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. Review PMID: 14608122 [PubMed - indexed for MEDLINE] 1391: J Nutr. 2003 Nov;133(11):3319-22. Biotechnology and food systems in developing countries. Timmer CP. Development Alternatives, Incorporated (DAI), Bethesda, MD 20814, USA. Peter_Timmer@dai.com Even in a world with adequate food supplies in global markets, which is the situation today, biotechnology offers important opportunities to developing countries in four domains. First, many agronomically hostile or degraded environments require major scientific breakthroughs to become productive agricultural systems. Few of these breakthroughs are likely to be achieved through traditional breeding approaches. Second, biofortification offers the promise of greater quantities and human availabilities of micronutrients from traditional staple foods, with obvious nutritional gains for poor consumers, especially their children. Third, many high yielding agricultural systems are approaching their agronomic potential. Radically new technologies will be required to sustain productivity growth in these systems, and only modern genetic technology offers this hope. Finally, many cropping systems use large quantities of chemical inputs, such as herbicides, pesticides and fertilizers that can be unhealthy for people and soils alike. Biotechnology offers the potential to reduce the need for these inputs in economically and environmentally sustainable ways. Applying these new technologies to society's basic foods raises obvious concerns for both human and ecological health. For some, these concerns have become outright fear, and this has mobilized a backlash against genetically modified foods in any form. These concerns (and fears) must be addressed carefully and rationally so that the public understands the risks (which are not zero) and benefits (which might be enormous). Only the scientific community has the expertise and credibility to build this public understanding. Publication Types: Lectures PMID: 14608038 [PubMed - indexed for MEDLINE] 1392: Orv Hetil. 2003 Sep 21;144(38):1853-60. [Etiopathogenesis of inflammatory bowel diseases] [Article in Hungarian] Lakatos L, Lakatos PL. Csolnoky Ferenc Kórház, I. sz. Belgyógyászati Osztály, Veszprém. lakatosvmkgastro@hotmail.com The pathogenesis of IBD is only partly understood; various environmental and host (e.g. genetic-, epithelial-, immune and non-immune) factors are involved. It is a multifactorial polygenic disease with probable genetic heterogeneity, some genes are associated with IBD itself, while others increase the risk of ulcerative colitis (UC) or Crohn's disease (CD) or are associated with disease location and/or behaviour. The role of environmental factors, in particular, enteric antigens, smoking and non-steroid anti-inflammatory drug use has been well established. However uptil now no proof of a role of any unique pathogenic bacteria or special dietary and/or psychosocial factor has been identified. In this hypothesis, the disease may develop in a genetically predisposed host as a consequence of disregulated immune response to environmental, in particular, enteric antigens, resulting in a continuous immune-mediated inflammation (in CD predominantly Th-1, in UC a modified Th-2 mechanisms are involved) and not in tolerance. As a consequence, the permeability of mucosa and the antigen challenge increases, in contrast, the disregulated immune response is unable to downregulate the inflammatory process. This will result in a continuous inflammation and tissue damage. The pathogenesis of CD is thought to be mainly an antigen driven, T-lymphocyte dependent process, while in UC the role of epithelial factors and activated granulocytes are essential. Publication Types: English Abstract Review PMID: 14596023 [PubMed - indexed for MEDLINE] 1393: Riv Biol. 2003 May-Aug;96(2):207-23. The ecological risks of transgenic plants. Giovannetti M. Department of Chemistry and Agricultural Biotechnology, University of Pisa, Via del Borghetto 80, 56124 Pisa, Italy. mgiova@agr.unipi.it Biotechnologies have been utilized "ante litteram" for thousands of years to produce food and drink and genetic engineering techniques have been widely applied to produce many compounds for human use, from insulin to other medicines. The debate on genetically modified (GM) organisms broke out all over the world only when GM crops were released into the field. Plant ecologists, microbiologists and population geneticists carried out experiments aimed at evaluating the environmental impact of GM crops. The most significant findings concern: the spread of transgenes through GM pollen diffusion and its environmental impact after hybridisation with closely related wild species or subspecies; horizontal gene transfer from transgenic plants to soil microbes; the impact of insecticide proteins released into the soil by transformed plants on non-target microbial soil communities. Recent developments in genetic engineering produced a technology, dubbed "Terminator", which protects patented genes introduced in transgenic plants by killing the seeds in the second generation. This genetic construct, which interferes so heavily with fundamental life processes, is considered dangerous and should be ex-ante evaluated taking into account the data on "unexpected events", as here discussed, instead of relying on the "safe until proven otherwise" claim. Awareness that scientists, biotechnologists and genetic engineers cannot answer the fundamental question "how likely is that transgenes will be transferred from cultivated plants into the natural environment?" should foster long-term studies on the ecological risks and benefits of transgenic crops. Publication Types: Review PMID: 14595899 [PubMed - indexed for MEDLINE] 1394: Nat Biotechnol. 2003 Nov;21(11):1274-9. A framework for designing transgenic crops--science, safety and citizen's concerns. König A. Harvard University, Harvard Center for Risk Analysis, 718 Huntington Avenue, Boston, Masschusetts 02115, USA. ariane_koenig@harvard.edu Publication Types: Research Support, Non-U.S. Gov't PMID: 14595351 [PubMed - indexed for MEDLINE] 1395: Nat Biotechnol. 2003 Nov;21(11):1257-8. GM confusion in Brazil. Bonalume Neto R. Publication Types: News PMID: 14595342 [PubMed - indexed for MEDLINE] 1396: Curr Biol. 2003 Oct 28;13(21):R819-20. Not yet a GM nation. Dixon B. Publication Types: News PMID: 14588251 [PubMed - indexed for MEDLINE] 1397: Health Policy. 2003 Nov;66(2):179-97. The precautionary principle within European Union public health policy. The implementation of the principle under conditions of supranationality and citizenship. Antonopoulou L, van Meurs P. Department of Economics, Aristotle University of Thessaloniki, Thessaloniki, Greece. The present study examines the precautionary principle within the parameters of public health policy in the European Union, regarding both its meaning, as it has been shaped by relevant EU institutions and their counterparts within the Member States, and its implementation in practice. In the initial section I concentrate on the methodological question of "scientific uncertainty" concerning the calculation of risk and possible damage. Calculation of risk in many cases justifies the adopting of preventive measures, but, as it is argued, the principle of precaution and its implementation cannot be wholly captured by a logic of calculation; such a principle does not only contain scientific uncertainty-as the preventive principle does-but it itself is generated as a principle by this scientific uncertainty, recognising the need for a society to act. Thus, the implementation of the precautionary principle is also a simultaneous search for justification of its status as a principle. This justification would result in the adoption of precautionary measures against risk although no proof of this principle has been produced based on the "cause-effect" model. The main part of the study is occupied with an examination of three cases from which the stance of the official bodies of the European Union towards the precautionary principle and its implementation emerges: the case of the "mad cows" disease, the case of production and commercialization of genetically modified foodstuffs. The study concludes with the assessment that the effective implementation of the precautionary principle on a European level depends on the emergence of a concerned Europe-wide citizenship and its acting as a mechanism to counteract the material and social conditions that pose risks for human health. PMID: 14585517 [PubMed - indexed for MEDLINE] 1398: J Biotechnol. 2003 Nov 6;105(3):227-33. Multiplex polymerase chain reaction/membrane hybridization assay for detection of genetically modified organisms. Su W, Song S, Long M, Liu G. School of Biotechnology, Jimei University, Shi-Gu Road, 9 Xiamen, Fujian 361021, China. To improve detection efficiency and result accuracy, four screening primer pairs, four identifying primer pairs, one common primer pair and corresponding probes were designed for the development of multiplex polymerase chain reaction/membrane hybridization assay (MPCR-MHA) for detection of the foreign genes insert in genetically modified organisms (GMOs). After detecting condition and parameter were optimized and determined, MPCR reactions were developed for amplifying several target genes simultaneously in one tube. Primers were labeled with biotin at the 5'-end; biotinylated MPCR products were detected by hybridization to the oligonucleotide probes immobilized on a membrane with subsequent colorimetric detection to confirm hybridization. The testing of screening primers can judge whether the sample contains GMOs, and that of identifying primers can further judge what kinds of trait genes are contained in the sample. We detected nine soybean samples, six maize samples, seven potato samples and two rice samples by the MPCR-MHA method; at the same time we also detected them with single PCR-MHA method. The results between two methods have good consistency. Publication Types: Research Support, Non-U.S. Gov't PMID: 14580794 [PubMed - indexed for MEDLINE] 1399: Biochem Biophys Res Commun. 2003 Nov 7;311(1):223-8. Genetic attachment of undecane peptides to ovomucoid third domain can suppress the production of specific IgG and IgE antibodies. Mine Y, Rupa P. Department of Food Science, University of Guelph, Ont., Canada. ymine@uoguelph.ca An undecane peptide (Gly-Ser-Pro-Gly-Ile-Pro-Gly-Ser-Thr-Gly-Met) was genetically attached to the N-terminus of ovomucoid third domain (DIII) to investigate structural characteristics of linear IgE and IgG (B cell) epitopes in DIII with respect to modulation of the immune response towards antigenicity and allergenicity. Balb/c mice were sensitized with native DIII, wild type recombinant DIII, and recombinant modified DIII containing the extra amino acid stretch. The immune responses to the antigens were compared using enzyme-linked immunosorbent assay. Interestingly, specific IgE and IgG levels were suppressed when the modified DIII was used as antigen. This was further confirmed by synthesizing immunodominant IgE and IgG epitopes of DIII on cellulose acetate membrane (SPOTs) and probing them with antibodies raised against DIII antigens. Anti-recombinant wild type DIII anti-serum showed strong binding activities to immunodominant IgE and IgG epitopes, while anti-modified DIII serum did not show any significant binding to the IgE and IgG epitopes. Thus, it is clearly demonstrated that the amino acid stretch in DIII is masking the immune reactive epitope. Genetical attachment of peptides into DIII was found to be effective in reducing the production of specific IgE and IgG antibodies in mice. Publication Types: Evaluation Studies Research Support, Non-U.S. Gov't PMID: 14575717 [PubMed - indexed for MEDLINE] 1400: J Theor Biol. 2003 Nov 21;225(2):241-55. Spatially explicit modelling of transgenic maize pollen dispersal and cross-pollination. Loos C, Seppelt R, Meier-Bethke S, Schiemann J, Richter O. Department of Environmental System Analysis, Institute of Geoecology, Technical University of Braunschweig, Langer Kamp 19c, 38106 Braunschweig, Germany. Modelling of pollen dispersal and cross-pollination is of great importance for the ongoing discussion on thresholds for the adventitious presence of genetically modified material in food and feed. Two different modelling approaches for pollen dispersal are used to simulate the cross-pollination rate of pollen emerged from an adjacent transgenic crop field. The models are applied to cross-pollination data from field experiments with transgenic maize (Zea mays). The data were generated by an experimental setup specifically designed to suit the demands of mathematical modelling. First a Gaussian plume model is used for the simulation of pollen transport in and from plant canopies. This is a semiempirical approach combining the atmospheric diffusion equation and Lagrangian methodology. The second model is derived from the localised near field (LNF) theory and based on the physical processes in the canopy. Both modelling approaches prove to be appropriate for the simulation of the cross-pollination rates at distances of about 7.5m and more from the transgene source. The simulation of the cross-pollination rate is less precise at the edge of the source plot especially with the LNF theory. However, the simulation results lie within the range of variability of the observations. Concluding can be pointed out that both models might be adapted to other pollen dispersal experiments of different crops and plot sizes. PMID: 14575658 [PubMed - indexed for MEDLINE] 1401: Nature. 2003 Oct 23;425(6960):751. Biosafety trials darken outlook for transgenic crops in Europe. Giles J. Publication Types: News PMID: 14574368 [PubMed - indexed for MEDLINE] 1402: Alcohol Clin Exp Res. 2003 Oct;27(10):1554-62. Hypericum perforatum CO2 extract and opioid receptor antagonists act synergistically to reduce ethanol intake in alcohol-preferring rats. Perfumi M, Santoni M, Cippitelli A, Ciccocioppo R, Froldi R, Massi M. Department of Pharmacological Sciences and Experimental Medicine, University of Camerino, Camerino (Macerata), Italy. marina.perfumi@unicam.it BACKGROUND: Hypericum perforatum extracts attenuate ethanol intake in alcohol-preferring rats. The opioid receptor antagonists, naloxone and naltrexone, reduce ethanol intake in rats and humans. The combination of different agents that reduce ethanol intake has been proposed as an approach to the pharmacotherapy of alcoholism. This study evaluated the effect on ethanol intake of the combined administration of a CO2 H. perforatum extract and naloxone or naltrexone in genetically selected Marchigian Sardinian alcohol-preferring rats. METHODS: Ten percent (v/v) ethanol intake was offered 2 hr per day at the beginning of the dark phase of the reverse light-dark cycle. H. perforatum CO2 extract was given intragastrically, 1 hr before access to ethanol. Naloxone or naltrexone was given by intraperitoneal injection 10 min before the extract. RESULTS: H. perforatum CO2 extract reduced ethanol intake at 31 or 125 mg/kg, but not 7 mg/kg. These doses neither modified food or water intake during access to ethanol, nor reduce 0.2% saccharin intake. Naloxone reduced ethanol and food intake at 3 or 5 mg/kg, but not 1 mg/kg. When naloxone 1 mg/kg was combined with the three doses of H. perforatum CO2 extract, the attenuation of ethanol intake was more pronounced than that observed after the administration of the extract alone. Alcohol intake was also significantly reduced by 7 mg/kg of H. perforatum CO2 extract combined with naloxone 1 mg/kg. The combined treatments never modified the rat's locomotor activity nor the simultaneous intake of food, water or 0.2% saccharin. Naltrexone reduced ethanol intake at 1 and 3 mg/kg, but not at 0.5 mg/kg. When naltrexone 0.5 mg/kg was combined with H. perforatum CO2 extract 7 mg/kg, ethanol intake was markedly reduced. CONCLUSIONS: These findings provide evidence that H. perforatum CO2 extract and opiate receptor antagonists act synergistically to induce a pronounced and selective reduction of voluntary ethanol consumption in alcohol-preferring rats. Publication Types: Comparative Study Research Support, Non-U.S. Gov't PMID: 14574225 [PubMed - indexed for MEDLINE] 1403: Protein Sci. 2003 Nov;12(11):2434-42. Identification of transglutaminase-mediated deamidation sites in a recombinant alpha-gliadin by advanced mass-spectrometric methodologies. Mazzeo MF, De Giulio B, Senger S, Rossi M, Malorni A, Siciliano RA. Centro di Spettrometria di Massa Proteomica e Biomolecolare, Istituto di Scienze dell'Alimentazione del CNR, Avellino, Italy. Celiac disease is a permanent immune-mediated food intolerance triggered by ingestion of wheat gliadins in genetically susceptible individuals. It has been reported that tissue transglutaminase plays an important role in the onset of celiac disease by converting specific glutamine residues within gliadin fragments into glutamic acid residues. This process increases binding affinity of gliadin peptides to HLA-DQ2/DQ8 molecules, thus enhancing the immune response. The aim of the present study was to achieve a detailed structural characterization of modifications induced by transglutaminase on gliadin peptides. Therefore, structural analyses were carried out on a recombinant alpha-gliadin and on a panel of 26 synthetic peptides, overlapping the complete protein sequence. Modified glutamine residues were identified by means of advanced mass-spectrometric methodologies on the basis of MALDI-TOF-MS and tandem mass spectrometry. Results led to the identification of 19 of 94 glutamine residues present in the recombinant alpha-gliadin, which were converted into glutamic acid residues by a transglutaminase-mediated reaction. This allowed us to achieve a global view of the modifications induced by the enzyme on this protein. Furthermore, results gathered could likely be utilized as relevant information for a better understanding of processes leading to T-cell recognition of gliadin peptides involved in celiac disease. Publication Types: Research Support, Non-U.S. Gov't PMID: 14573857 [PubMed - indexed for MEDLINE] 1404: Trends Biotechnol. 2003 Nov;21(11):491-7. Genetically modified probiotics in foods. Ahmed FE. Department of Radiation Oncology, LSB 014, Leo W. Jenkins Cancer Center, The Brody School of Medicine, East Carolina University, Greenville, NC 27858, USA. ahmedf@mail.ecu.edu Probiotics have many potential therapeutic uses, but have not been universally accepted because of a lack of understanding of their action. Lactic acid bacteria (LAB) have been modified by traditional and genetic engineering methods to produce new varieties. Modern techniques of molecular biology have facilitated the identification of probiotic LAB strains, but only a few LAB have been modified by recombinant-DNA technology because of consumer resistance to their introduction to markets, especially in Europe. Publication Types: Review PMID: 14573362 [PubMed - indexed for MEDLINE] 1405: Environ Sci Technol. 2003 Oct 1;37(19):343A. GM products: at least a label. Schnoor JL. est@uiowa.edu PMID: 14572070 [PubMed - indexed for MEDLINE] 1406: Biotechnol Lett. 2003 Sep;25(18):1485-90. Genetically modified soybeans: false-positive detection in fermented natural soybean (tempe). Prakoso B, Nitisinprasert S, Stevens WF. Bioprocess Technology, Asian Institute of Technology, Thailand. Tempe was prepared using mixtures of natural soybean and genetically modified Roundup Ready (RUR) soybean fermented with natural Rhizopus sp. The amount of RUR soybean was quantified using an ELISA plate test. The RUR signal decreased during fermentation. In the control experiments on fermentation of non-RUR soybean, the tempe gave a false-positive RUR signal. The cross-reacting substance was generated only in non-RUR soybean during fermentation by Rhizopus sp., Rhizopus oligosporus, R. oryzae, Mucor rouxii and Aspergillus awamori. Publication Types: Comparative Study Evaluation Studies Validation Studies PMID: 14571970 [PubMed - indexed for MEDLINE] 1407: Science. 2003 Oct 17;302(5644):357. Comment in: Science. 2004 Jan 16;303(5656):310. Agriculture and the developing world. Kennedy D. Publication Types: Editorial PMID: 14563974 [PubMed - indexed for MEDLINE] 1408: Nature. 2003 Oct 16;425(6959):656-7. Damned if they do, damned if they don't... Giles J. Publication Types: News PMID: 14562072 [PubMed - indexed for MEDLINE] 1409: Nature. 2003 Oct 16;425(6959):655. Time to choose. Aldhous P. Publication Types: News PMID: 14562071 [PubMed - indexed for MEDLINE] 1410: Philos Trans R Soc Lond B Biol Sci. 2003 Nov 29;358(1439):1899-913. Responses of plants and invertebrate trophic groups to contrasting herbicide regimes in the Farm Scale Evaluations of genetically modified herbicide-tolerant crops. Hawes C, Haughton AJ, Osborne JL, Roy DB, Clark SJ, Perry JN, Rothery P, Bohan DA, Brooks DR, Champion GT, Dewar AM, Heard MS, Woiwod IP, Daniels RE, Young MW, Parish AM, Scott RJ, Firbank LG, Squire GR. Scottish Crop Research Institute, Invergowrie, Dundee DD2 5DA, UK. chawes@scri.sari.ac.uk Effects of genetically modified herbicide-tolerant (GMHT) and conventional crop management on invertebrate trophic groups (herbivores, detritivores, pollinators, predators and parasitoids) were compared in beet, maize and spring oilseed rape sites throughout the UK. These trophic groups were influenced by season, crop species and GMHT management. Many groups increased twofold to fivefold in abundance between early and late summer, and differed up to 10-fold between crop species. GMHT management superimposed relatively small (less than twofold), but consistent, shifts in plant and insect abundance, the extent and direction of these effects being dependent on the relative efficacies of comparable conventional herbicide regimes. In general, the biomass of weeds was reduced under GMHT management in beet and spring oilseed rape and increased in maize compared with conventional treatments. This change in resource availability had knock-on effects on higher trophic levels except in spring oilseed rape where herbivore resource was greatest. Herbivores, pollinators and natural enemies changed in abundance in the same directions as their resources, and detritivores increased in abundance under GMHT management across all crops. The result of the later herbicide application in GMHT treatments was a shift in resource from the herbivore food web to the detritivore food web. The Farm Scale Evaluations have demonstrated over 3 years and throughout the UK that herbivores, detritivores and many of their predators and parasitoids in arable systems are sensitive to the changes in weed communities that result from the introduction of new herbicide regimes. Publication Types: Comparative Study Research Support, Non-U.S. Gov't PMID: 14561321 [PubMed - indexed for MEDLINE] 1411: Philos Trans R Soc Lond B Biol Sci. 2003 Nov 29;358(1439):1779-99. On the rationale and interpretation of the Farm Scale Evaluations of genetically modified herbicide-tolerant crops. Squire GR, Brooks DR, Bohan DA, Champion GT, Daniels RE, Haughton AJ, Hawes C, Heard MS, Hill MO, May MJ, Osborne JL, Perry JN, Roy DB, Woiwod IP, Firbank LG. Scottish Crop Research Institute, Invergowrie, Dundee DD2 5DA, UK. g.squire@scri.sari.ac.uk Farmland biodiversity and food webs were compared in conventional and genetically modified herbicide-tolerant (GMHT) crops of beet (Beta vulgaris L.), maize (Zea mays L.) and both spring and winter oilseed rape (Brassica napus L.). GMHT and conventional varieties were sown in a split-field experimental design, at 60-70 sites for each crop, spread over three starting years beginning in 2000. This paper provides a background to the study and the rationale for its design and interpretation. It shows how data on environment, field management and the biota are used to assess the current state of the ecosystem, to define the typical arable field and to devise criteria for selecting, sampling and auditing experimental sites in the Farm Scale Evaluations. The main functional and taxonomic groups in the habitat are ranked according to their likely sensitivity to GMHT cropping, and the most responsive target organisms are defined. The value of the seedbank as a baseline and as an indicator of historical trends is proposed. Evidence from experiments during the twentieth century is analysed to show that large changes in field management have affected sensitive groups in the biota by ca. 50% during a year or short run of years--a figure against which to assess any positive or negative effects of GMHT cropping. The analysis leads to a summary of factors that were, and were not, examined in the first 3 years of the study and points to where modelling can be used to extrapolate the effects to the landscape and the agricultural region. Publication Types: Comparative Study Evaluation Studies Research Support, Non-U.S. Gov't PMID: 14561314 [PubMed - indexed for MEDLINE] 1412: Mol Plant Microbe Interact. 2003 Oct;16(10):936-44. Cucumber mosaic virus infection transiently breaks dsRNA-induced transgenic immunity to Potato virus Y in tobacco. Mitter N, Sulistyowati E, Dietzgen RG. Queensland Department of Primary Industries, Agency for Food and Fibre Sciences, Agricultural Biotechnology and Cooperative Research Centre for Tropical Plant Protection, The University of Queensland, St. Lucia Qld 4072, Australia. Post-transcriptional gene silencing (PTGS), an intrinsic plant defense mechanism, can be efficiently triggered by double stranded (ds)RNA-producing transgenes and can provide high level virus resistance by specific targeting of cognate viral RNA. The discovery of virus-encoded suppressors of PTGS led to concerns about the stability of such resistance. Here, we show that Cucumber mosaic virus (CMV) is able to suppress dsRNA-induced PTGS and the associated Potato virus Y (PVY) immunity in tobacco. CMV suppression supported only a transient PVY accumulation and did not prevent recovery of the transgenic plants from PVY infection. CMV inoculation resulted in strongly increased transgene mRNA levels due to suppression of PTGS, but accumulation of PVY-specific small interfering (si)RNA was unaffected. However, PVY accumulation in previously immune plants resulted in increased PVY siRNA levels and transgene mRNA was no longer detected, despite the presence of CMV. Transgene mRNA returned to high levels once PVY was no longer detected in CMV-infected plants. Recovered and chronically CMV-infected tissues were immune to further PVY infection. Publication Types: Research Support, Non-U.S. Gov't PMID: 14558695 [PubMed - indexed for MEDLINE] 1413: Mol Genet Genomics. 2003 Dec;270(4):362-8. Epub 2003 Oct 11. Uptake of amplifiable fragments of retrotransposon DNA from the human alimentary tract. Forsman A, Ushameckis D, Bindra A, Yun Z, Blomberg J. Section of Virology, Department of Medical Sciences, Academic Hospital, Uppsala University, 751 85 Uppsala, Sweden. Few attempts have been made to study the transfer of DNA from ingested food across the intestinal barrier. A low uptake of ingested DNA has been observed in mice, cattle and poultry. There have been no reports on humans so far. Maintenance of species barriers, protection against retrotransposons, optimisation of oral DNA vaccines and the fate of genetically modified foodstuffs are issues where this topic is of importance. We therefore used the high-copy-number rabbit retrotransposon RERV-H, and rabbit mitochondrial DNA, to study the transfer of DNA from ingested rabbit meat into the bloodstream of two human volunteers. A quantitative PCR was used to measure RERV-H levels in food and in the blood. Amplification with the primers selected results in the generation of a 250-bp fragment of RERV-H. Transfer across the intestinal epithelium could be demonstrated in both subjects. Levels of the fragment in the bloodstream peaked at 1-3 h after ingestion of the experimental meal. One hour after a meal of rabbit meat containing 10(14) copies of RERV-H DNA, a maximum concentration of 200 copies of RERV-H DNA per ml of peripheral blood was observed, which corresponds to the uptake of approximately 10(6) RERV-H DNA copies in 1 h. RERV-H DNA was detected in both cellular and plasma compartments. Both rabbit retrotransposon and mitochondrial DNA was taken up from the human alimentary tract. The size of the fragments detected is similar to that of SINE retrotransposons (approximately 300 bp). The fate and functionality of alimentary DNA in humans will require further study. Publication Types: Research Support, Non-U.S. Gov't PMID: 14556071 [PubMed - indexed for MEDLINE] 1414: PLoS Biol. 2003 Oct;1(1):E8. Epub 2003 Oct 13. Genetically modified corn--environmental benefits and risks. Gewin V. gewin@nasw.org PMID: 14551906 [PubMed - indexed for MEDLINE] 1415: Appetite. 2003 Oct;41(2):207-9. Consumer perceptions of genetically modified and organic foods. What kind of knowledge matters? Koivisto Hursti UK, Magnusson MK. Department of Public Health and Caring Sciences, Section of Caring Sciences, Uppsala University, Uppsala Science Park, 751 83 Uppsala, Sweden. ulla-kaisa.koivisto@pubcare.uu.se Publication Types: Research Support, Non-U.S. Gov't PMID: 14550321 [PubMed - indexed for MEDLINE] 1416: Biotechnol Adv. 1999 Dec 30;17(8):647-78. Biotechnology products and European consumers. Moses V. Division of Life Sciences, King's College, London, SE1 8WA, UK. v.moses@qmw.ac.uk More than 100 interviews conducted during 1997 with European food manufacturers and retailers, trade associations, government departments, consumer groups, environmental organizations and some individual academic scientists revealed how differences in the perceived attitudes of consumers gave rise to varying approaches by suppliers to the possible introduction of transgenic foods. European consumers generally are not against the pharmaceutical products of biotechnology but are much less willing to accept food and food ingredients, especially when derived from genetically modified plants. Objections are mainly based on fears for the health and safety of the consumer, worries about the possibility of deleterious effects on the environment, and a range of moral and ethical concerns often deriving from a distaste, however expressed, at the concept of interfering with nature. Consumer understanding of the science underlying biotechnology is patchy; in no country does more than a small proportion of the population claim a good grasp. Partly no doubt as a consequence of these attitudes, the introduction of genetically modified foods into Europe has occurred slowly and, during the period of this study, perhaps only in the Netherlands and the UK. PMID: 14538121 [PubMed] 1417: Biotechnol Adv. 2000 May;18(3):179-206. The impact of genetic modification of human foods in the 21st century: a review. Uzogara SG. Bioanalytical-PK Department, Alkermes Inc., Cambridge, MA 02139, USA. uzogara_stella@alkermes.com Genetic engineering of food is the science which involves deliberate modification of the genetic material of plants or animals. It is an old agricultural practice carried on by farmers since early historical times, but recently it has been improved by technology. Many foods consumed today are either genetically modified (GM) whole foods, or contain ingredients derived from gene modification technology. Billions of dollars in U.S. food exports are realized from sales of GM seeds and crops. Despite the potential benefits of genetic engineering of foods, the technology is surrounded by controversy. Critics of GM technology include consumer and health groups, grain importers from European Union (EU) countries, organic farmers, environmentalists, concerned scientists, ethicists, religious rights groups, food advocacy groups, some politicians and trade protectionists. Some of the specific fears expressed by opponents of GM technology include alteration in nutritional quality of foods, potential toxicity, possible antibiotic resistance from GM crops, potential allergenicity and carcinogenicity from consuming GM foods. In addition, some more general concerns include environmental pollution, unintentional gene transfer to wild plants, possible creation of new viruses and toxins, limited access to seeds due to patenting of GM food plants, threat to crop genetic diversity, religious, cultural and ethical concerns, as well as fear of the unknown. Supporters of GM technology include private industries, research scientists, some consumers, U.S. farmers and regulatory agencies. Benefits presented by proponents of GM technology include improvement in fruit and vegetable shelf-life and organoleptic quality, improved nutritional quality and health benefits in foods, improved protein and carbohydrate content of foods, improved fat quality, improved quality and quantity of meat, milk and livestock. Other potential benefits are: the use of GM livestock to grow organs for transplant into humans, increased crop yield, improvement in agriculture through breeding insect, pest, disease, and weather resistant crops and herbicide tolerant crops, use of GM plants as bio-factories to yield raw materials for industrial uses, use of GM organisms in drug manufacture, in recycling and/or removal of toxic industrial wastes. The potential risks and benefits of the new technology to man and the environment are reviewed. Ways of minimizing potential risks and maximizing the benefits of GM foods are suggested. Because the benefits of GM foods apparently far outweigh the risks, regulatory agencies and industries involved in GM food business should increase public awareness in this technology to enhance worldwide acceptability of GM foods. This can be achieved through openness, education, and research. PMID: 14538107 [PubMed] 1418: Biotechnol Adv. 2001 Nov;19(7):539-54. Management strategies for agricultural biotechnology in small countries. A case study of Israel. Shalhevet S, Haruvy N, Spharim I. Department of Economics, Agricultural Research Organization, Bet Dagan, Israel. sarit@agri.gov.il Agricultural biotechnology is concentrated in four major countries. This paper suggests strategies for developing it in small countries, based on analysis of the world trends and the characteristics of small countries. Israel is presented as a specific case study. The main relevant trends are domination by big companies, consumer concerns on genetically modified foods, and focusing on consumer benefits and specific market niches. Small countries' disadvantages include companies that are too small to benefit fully from research, difficulty in raising funds, lack of infrastructures and experienced management personnel, and public sector research organizations that are unsuitable for commercializing research. The recommended strategies include: developing a large number of low-volume products and small market niches, forming partnerships with intermediaries (such as food companies), specializing in intermediate products (such as the seed or the gene patent), and conducting market research and cost-benefit analysis in advance. Additional strategies include developing benefits that are unique to genetically modified foods and focusing on benefits specifically for consumers who accept genetically modified foods, rather than on benefits for the average consumer. A national representative organization could buy and rent out expensive equipment, finance specific projects in return for the commercial rights, and perform collective marketing research and marketing. Israel has the advantages of a successful agricultural sector and complementary scientific research, and should focus on those fruits, vegetables, and flowers for which it already has the experience and infrastructure. PMID: 14538065 [PubMed] 1419: Environ Sci Pollut Res Int. 2003;10(5):273-6. Biotechnology for food, energy, and industrial products: new opportunities for bio-based products. Young AL. Institute for Science and Public Policy, Sarkeys Energy Center, The University of Oklahoma, 100 East Boyd, Room 510, Norman, Oklahoma, 73019, USA. youngrisk@attbi.com Publication Types: Review PMID: 14535638 [PubMed - indexed for MEDLINE] 1420: Vet Res Commun. 2003 Sep;27 Suppl 1:699-701. Detection of genetically modified organisms (GMOs) in food and feedstuff. Novelli E, Balzan S, Segato S, De Rigo L, Ferioli M. Dipartimento di Sanità Pubblica, Patologia Comparata e Igiene Veterinaria, Università degli Studi di Padova, 35020 Legnaro, Italy. enrico.novelli@unipd.it PMID: 14535501 [PubMed - indexed for MEDLINE] 1421: Vet Res Commun. 2003 Sep;27 Suppl 1:659-61. Antigenic recombinant proteins expressed in tobacco seeds as a model for edible vaccines against swine oedema. Rossi L, Baldi A, Dell'Orto V, Fogher C. Department of Veterinary Sciences and Technology for Food Safety, Faculty of Veterinary Medicine, University of Milan, Via Celoria 10, 20133, Milano, Italy. PMID: 14535491 [PubMed - indexed for MEDLINE] 1422: Nature. 2003 Oct 9;425(6958):559. Comment in: Nature. 2003 Dec 4;426(6966):495. Flaws undermine results of UK biotech debate. Campbell S, Townsend E. Publication Types: Letter PMID: 14534562 [PubMed - indexed for MEDLINE] 1423: Appl Environ Microbiol. 2003 Oct;69(10):6121-7. Marker rescue studies of the transfer of recombinant DNA to Streptococcus gordonii in vitro, in foods and gnotobiotic rats. Kharazmi M, Sczesny S, Blaut M, Hammes WP, Hertel C. Institute of Food Technology, University of Hohenheim, Stuttgart, Germany. A plasmid marker rescue system based on restoration of the nptII gene was established in Streptococcus gordonii to study the transfer of bacterial and transgenic plant DNA by transformation. In vitro studies revealed that the marker rescue efficiency depends on the type of donor DNA. Plasmid and chromosomal DNA of bacteria as well as DNA of transgenic potatoes were transferred with efficiencies ranging from 8.1 x 10(-6) to 5.8 x 10(-7) transformants per nptII gene. Using a 792-bp amplification product of nptII the efficiency was strongly decreased (9.8 x 10(-9)). In blood sausage, marker rescue using plasmid DNA was detectable (7.9 x 10(-10)), whereas in milk heat-inactivated horse serum (HHS) had to be added to obtain an efficiency of 2.7 x 10(-11). No marker rescue was detected in extracts of transgenic potatoes despite addition of HHS. In vivo transformation of S. gordonii LTH 5597 was studied in monoassociated rats by using plasmid DNA. No marker rescue could be detected in vivo, although transformation was detected in the presence of saliva and fecal samples supplemented with HHS. It was also shown that plasmid DNA persists in rat saliva permitting transformation for up to 6 h of incubation. It is suggested that the lack of marker rescue is due to the absence of competence-stimulating factors such as serum proteins in rat saliva. Publication Types: Research Support, Non-U.S. Gov't PMID: 14532070 [PubMed - indexed for MEDLINE] 1424: Nat Rev Genet. 2003 Oct;4(10):839-43. GM crops: science, politics and communication. Arntzen CJ, Coghlan A, Johnson B, Peacock J, Rodemeyer M. Center for Production of Vacccines from Applied Crop Science, Arizona Biodesign Institute at Arizona State University, United States. As the public debate in Europe about genetically modified (GM) crops heats up and the trade row between the United States and the European Union over GM food escalates, what better time to examine the issues with an international group of experts (Box 1). Their views are diverse, but they all agree that we need more impartial communication, less propaganda and an effective regulatory regime that is based on a careful case-by-case consideration of GM technology. It seems that GM crops are here to stay, so let us hope that these requirements are met and that the developing nations that perhaps have the most to gain from this technology can start to reap its benefits. Publication Types: Review PMID: 14526380 [PubMed - indexed for MEDLINE] 1425: Curr Biol. 2003 Sep 30;13(19):R745-6. Italy sets new hurdles for GM crops. Williams N. Publication Types: News PMID: 14521845 [PubMed - indexed for MEDLINE] 1426: Planta. 2003 Jul;217(3):367-73. Epub 2003 Mar 18. Generation of cyanogen-free transgenic cassava. Siritunga D, Sayre RT. Department of Plant Biology, The Ohio State University, Columbus 43210, USA. Cassava ( Manihot esculenta Crantz.) is the major source of calories for subsistence farmers in sub-Saharan Africa. Cassava, however, contains potentially toxic levels of the cyanogenic glucoside, linamarin. The cyanogen content of cassava foods can be reduced to safe levels by maceration, soaking, rinsing and baking; however, short-cut processing techniques can yield toxic food products. Our objective was to eliminate cyanogens from cassava so as to eliminate the need for food processing. To achieve this goal we generated transgenic acyanogenic cassava plants in which the expression of the cytochrome P450 genes ( CYP79D1 and CYP79D2), that catalyze the first-dedicated step in linamarin synthesis, was inhibited. Using a leaf-specific promoter to drive the antisense expression of the CYP79D1/ CYP79D2 genes we observed up to a 94% reduction in leaf linamarin content associated with an inhibition of CYP79D1 and CYP79D2 expression. Importantly, the linamarin content of roots also was reduced by 99% in transgenic plants having between 60 and 94% reduction in leaf linamarin content. Analysis of CYP79D1/ CYP79D2 transcript levels in transgenic roots indicated they were unchanged relative to wild-type plants. These results suggest that linamarin is transported from leaves to roots and that a threshold level of leaf linamarin production is required for transport. Publication Types: Research Support, Non-U.S. Gov't PMID: 14520563 [PubMed - indexed for MEDLINE] 1427: Mod Law Rev. 1998 Sep;61(5):621-60. Regulation as facilitation: negotiating the genetic revolution. Black J. Law Department, London School of Economics and Political Science, UK. PMID: 14518446 [PubMed - indexed for MEDLINE] 1428: Plant Physiol. 2003 Oct;133(2):589-96. Epub 2003 Sep 25. Erratum in: Plant Physiol. 2005 Oct;139(2):1095. Functional expression of a bacterial heavy metal transporter in Arabidopsis enhances resistance to and decreases uptake of heavy metals. Lee J, Bae H, Jeong J, Lee JY, Yang YY, Hwang I, Martinoia E, Lee Y. National Research Laboratory of Phytoremediation, Division of Molecular Life Sciences, Pohang University of Science and Technology, Pohang 790-784, Korea. Large parts of agricultural soil are contaminated with lead (Pb) and cadmium (Cd). Although most environments are not heavily contaminated, the low levels observed nonetheless pose a high risk of heavy metal accumulation in the food chain. Therefore, approaches to develop plants with reduced heavy metal uptake are important. Recently, many transgenic plants with increased heavy metal resistance and uptake of heavy metals were developed for the purpose of phytoremediation. However, to reduce heavy metal in the food chain, plants that transfer less heavy metals to the shoot are required. We tested whether an Escherichia coli gene, ZntA, which encodes a Pb(II)/Cd(II)/Zn(II) pump, could be useful for developing plants with reduced heavy metal content. Yeast cells transformed with this gene had improved resistance to Pb(II) and Cd(II). In Arabidopsis plants transformed with ZntA, ZntA was localized at the plasma membrane and improved the resistance of the plants to Pb(II) and Cd(II). The shoots of the transgenic plants had decreased Pb and Cd content. Moreover, the transgenic protoplasts showed lower accumulation of Cd and faster release of preloaded Cd than wild-type protoplasts. These results show that a bacterial transporter gene, ZntA, can be functionally expressed in plant cells, and that that it may be useful for the development of crop plants that are safe from heavy metal contamination. Publication Types: Research Support, Non-U.S. Gov't PMID: 14512517 [PubMed - indexed for MEDLINE] 1429: Trends Biotechnol. 2003 Oct;21(10):439-44. Comparative safety assessment for biotech crops. Kok EJ, Kuiper HA. RIKILT Institute for Food Safety, Bornsesteeg 45, PO Box 230, 6700 AE Wageningen, The Netherlands. esther.kok@wur.nl Publication Types: Research Support, Non-U.S. Gov't Review PMID: 14512230 [PubMed - indexed for MEDLINE] 1430: Arch Pediatr. 2003 May;10 Suppl 1:30s-31s. [Risk of chemical contaminants: from xenobiotics to genetically modified organisms] [Article in French] Vidailhet M. Pédiatrie 3 et génétique clinique, hôpital d'Enfants, rue du Morvan, 54511 Vandoeuvre, France. m.vidailhet@chu-nancy.fr PMID: 14509731 [PubMed - indexed for MEDLINE] 1431: Nature. 2003 Sep 25;425(6956):331. UK public opposes government on transgenic crops. Giles J. Publication Types: News PMID: 14508445 [PubMed - indexed for MEDLINE] 1432: Nature. 2003 Sep 25;425(6956):329. Dealing with democracy. [No authors listed] Publication Types: Editorial PMID: 14508443 [PubMed - indexed for MEDLINE] 1433: Best Pract Res Clin Gastroenterol. 2003 Oct;17(5):861-76. Genetically engineered probiotics. Steidler L. Department of Medicine, Alimentary Pharmabiotic Center, University College Cork, Western Road, Cork, Ireland. l.steidler@ucc.ie Probiotic micro-organisms have been used for many years. Originating as food supplements, they are now most often administered orally and offer an attractive alternative for treating of intestinal disorders. A better understanding of the mechanisms by which these micro-organisms act has now opened up possibilities for designing new probiotic strains. Through genetic engineering, it is possible not only to strengthen the effects of existing strains, but also to create completely new probiotics. These need not necessarily be composed only of bacterial products but can also include elements of regulatory systems or enzymes derived from a foreign-human-source. If designed carefully and with absolute attention to biological safety in its broadest sense, the development of genetically modified probiotics has the potential to revolutionize alimentary health. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 14507594 [PubMed - indexed for MEDLINE] 1434: Proc Nutr Soc. 2003 May;62(2):403-11. Micronutrient fortification of plants through plant breeding: can it improve nutrition in man at low cost? Bouis HE. International Food Policy Research Institute, 2033 K St NW, Washington DC 20006, USA. h.bouis@cgiar.org Can commonly-eaten food staple crops be developed that fortify their seeds with essential minerals and vitamins? Can farmers be induced to grow such varieties? If so, would this result in a marked improvement in human nutrition at a lower cost than existing nutrition interventions? An interdisciplinary international effort is underway to breed for mineral- and vitamin-dense varieties of rice, wheat, maize, beans and cassava for release to farmers in developing countries. The biofortification strategy seeks to take advantage of the consistent daily consumption of large amounts of food staples by all family members, including women and children as they are most at risk for micronutrient malnutrition. As a consequence of the predominance of food staples in the diets of the poor, this strategy implicitly targets low-income households. After the one-time investment is made to develop seeds that fortify themselves, recurrent costs are low and germplasm may be shared internationally. It is this multiplier aspect of plant breeding across time and distance that makes it so cost-effective. Once in place, the biofortified crop system is highly sustainable. Nutritionally-improved varieties will continue to be grown and consumed year after year, even if government attention and international funding for micronutrient issues fades. Biofortification provides a truly feasible means of reaching malnourished populations in relatively remote rural areas, delivering naturally-fortified foods to population groups with limited access to commercially-marketed fortified foods that are more readily available in urban areas. Biofortification and commercial fortification are, therefore, highly complementary. Breeding for higher trace mineral density in seeds will not incur a yield penalty. Mineral-packed seeds sell themselves to farmers because, as recent research has shown, these trace minerals are essential in helping plants resist disease and other environmental stresses. More seedlings survive and initial growth is more rapid. Ultimately, yields are higher, particularly in trace mineral-'deficient' soils in arid regions. Publication Types: Review PMID: 14506888 [PubMed - indexed for MEDLINE] 1435: Proc Nutr Soc. 2003 May;62(2):301-9. Engineering plants for animal feed for improved nutritional value. Williams PE. Syngenta Crop Protection AG, WRO-1002.13.63, Schwarzwaldallee 215, CH-4058 Basel, Switzerland. peter.williams@syngenta.com Feed formulation to meet nutritional requirements of livestock is becoming increasingly challenging. Regulations have banned the use of traditional high-quality protein supplements such as meat-and-bone meal, pollution from animal excreta of N and P is an issue and antibiotics are no longer available as insurance against the impact of enteric infection and feed anti-nutritional factors. The improved genetic potential of livestock is increasing daily requirement for energy and protein (essential amino acids). To benefit from the enhanced growth potential of livestock diets with high nutrient density are needed that can be formulated from crops without increased cost. Genetic modification of commodity crops used to manufacture animal feed in order to improve the density and quality of available nutrients is a potential solution to some of these problems. Furthermore, crops may be used as biofactories to produce molecules and products used in animal feed with considerable reductions in manufacturing fixed costs. Nevertheless, there are considerable not insurmountable challenges, such as the creation of sufficient economic value to deliver benefit to all members in the feed production chain, which is an essential element of identity preserving and delivering the technology to livestock producers. Individual output traits in the major commodity crops may not provide sufficient value to adequately compensate all the members of the feed production chain. Successful adoption of output traits may rely on inserting combinations of agronomic input traits with specific quality traits or increasing the value proposition by inserting combinations of output traits. Publication Types: Lectures PMID: 14506877 [PubMed - indexed for MEDLINE] 1436: Genetics. 2003 Sep;165(1):387-97. A new opaque variant of maize by a single dominant RNA-interference-inducing transgene. Segal G, Song R, Messing J. Waksman Institute, Rutgers, The State University of New Jersey, Piscataway, New Jersey 08854, USA. In maize, alpha-zeins, the main protein components of seed stores, are major determinants of nutritional imbalance when maize is used as the sole food source. Mutations like opaque-2 (o2) are used in breeding varieties with improved nutritional quality. However, o2 works in a recessive fashion by affecting the expression of a subset of 22-kD alpha-zeins, as well as additional endosperm gene functions. Thus, we sought a dominant mutation that could suppress the storage protein genes without interrupting O2 synthesis. We found that maize transformed with RNA interference (RNAi) constructs derived from a 22-kD zein gene could produce a dominant opaque phenotype. This phenotype segregates in a normal Mendelian fashion and eliminates 22-kD zeins without affecting the accumulation of other zein proteins. A system for regulated transgene expression generating antisense RNA also reduced the expression of 22-kD zein genes, but failed to give an opaque phenotype. Therefore, it appears that small interfering RNAs not only may play an important regulatory role during plant development, but also are effective genetic tools for dissecting the function of gene families. Since the dominant phenotype is also correlated with increased lysine content, the new mutant illustrates an approach for creating more nutritious crop plants. Publication Types: Research Support, U.S. Gov't, Non-P.H.S. PMID: 14504244 [PubMed - indexed for MEDLINE] 1437: QJM. 2003 Oct;96(10):779-80. Before Frankenstein. Berry C. PMID: 14500868 [PubMed - indexed for MEDLINE] 1438: Lancet. 2003 Sep 6;362(9386):835. Comment on: Lancet. 2003 Jun 14;361(9374):2051. Genetically modified crops in developing countries. Watts T. Publication Types: Comment Letter PMID: 13678895 [PubMed - indexed for MEDLINE] 1439: Nahrung. 2003 Aug;47(4):269-73. Assessment of protein nutritional quality and effects of traditional processes: a comparison between Ethiopian quality protein maize and five Ethiopian adapted normal maize cultivars. Fufa H, Akalu G, Wondimu A, Taffesse S, Gebre T, Schlosser K, Noetzold H, Henle T. Ethiopian Health and Nutrition Research Institute, PO Box 1242, Addis Abeba, Ethiopia. hfufa@yahoo.com The present study was designed to quantitatively measure and compare the levels and variations of total protein, individual amino acids, and computed protein efficiency ratio (C-PER) in raw and traditionally processed products of one recently released quality protein maize (QPM BH542) with four high-yield maize hybrids, namely flint BH660, semi-dent BH140, Pioneer 30H83, and Pioneer 30G97, as well as one local maize cultivar. The total protein content was variable among the cultivars ranging from 7% for BH660 to 8.6% for Pioneer 30H83, 8.9% for BH140, 9.8% for QPM BH542, 10.1% for local maize cultivar, and 11.8% for Pioneer 30G97, respectively. However, the QPM BH542 maize protein proved to be higher in nutritional quality than common maize proteins because it contained 30% to 82% more lysine, higher levels of arginine, tryptophan, histidine, threonine, cysteine, and valine. As a result, the QPM BH542 amino acid profile gives a good balance of total essential amino acids, limited only in lysine, and has a C-PER ratio of 2.2 compared to 1.14, 1.2, 1.4, 1.66, and 1.67 for Pioneer 30G97, local, BH-140, BH660, and Pioneer 30H83, respectively. The various traditional processes of maize have no significant effect on the protein nutritional quality of the new quality protein maize. Hence, the widely dissemination of it in agricultural cultivation as well as consumption by the general population is recommended. Publication Types: Comparative Study Research Support, Non-U.S. Gov't PMID: 13678268 [PubMed - indexed for MEDLINE] 1440: J Agric Food Chem. 2003 Sep 24;51(20):5829-34. Reliable detection and identification of genetically modified maize, soybean, and canola by multiplex PCR analysis. James D, Schmidt AM, Wall E, Green M, Masri S. Centre for Plant Health, Canadian Food Inspection Agency, 8801 East Saanich Road, Sidney, British Columbia, Canada V8L 1H3. Multiplex PCR procedures were developed for simultaneously detecting multiple target sequences in genetically modified (GM) soybean (Roundup Ready), maize (event 176, Bt11, Mon810, T14/25), and canola (GT73, HCN92/28, MS8/RF3, Oxy 235). Internal control targets (invertase gene in corn, lectin and beta-actin genes in soybean, and cruciferin gene in canola) were included as appropriate to assess the efficiency of all reactions, thereby eliminating any false negatives. Primer combinations that allowed the identification of specific lines were used. In one system of identification, simultaneous amplification profiling (SAP), rather than target specific detection, was used for the identification of four GM maize lines. SAP is simple and has the potential to identify both approved and nonapproved GM lines. The template concentration was identified as a critical factor affecting efficient multiplex PCRs. In canola, 75 ng of DNA template was more effective than 50 ng of DNA for the simultaneous amplification of all targets in a reaction volume of 25 microL. Reliable identification of GM canola was achieved at a DNA concentration of 3 ng/microL, and at 0.1% for GM soybean, indicating high levels of sensitivity. Nonspecific amplification was utilized in this study as a tool for specific and reliable identification of one line of GM maize. The primer cry1A 4-3' (antisense primer) recognizes two sites on the DNA template extracted from GM transgenic maize containing event 176 (European corn borer resistant), resulting in the amplification of products of 152 bp (expected) and 485 bp (unexpected). The latter fragment was sequenced and confirmed to be Cry1A specific. The systems described herein represent simple, accurate, and sensitive GMO detection methods in which only one reaction is necessary to detect multiple GM target sequences that can be reliably used for the identification of specific lines of GMOs. PMID: 13129280 [PubMed - indexed for MEDLINE] 1441: Biomed Environ Sci. 2003 Jun;16(2):149-56. Purification and immunity analysis of recombinant 6His-HPT protein expressed in E. coli. Yang LC, Zhu Z, Yang XG. Key Laboratory of Trace Element Nutrition of the Ministry of Health, China, Institute of Nutrition and Food Safety, Chinese Center for Diseases Control and Prevention, Beijing 100050, China. OBJECTIVE: To obtain HPT protein (Hygromycin B Phosphotransferase), a kind of plant selective maker gene product expressed from E. coli and to prepare the polyclonal antibody (pAbs) against it. METHODS: HPT cDNA fragment was obtained by PCR and was inserted into the prokaryotic expressing vector pBV222. Then the constructed recombinant plasmid pBV222-HPT was transferred into E. coli DH5alpha for HPT expression. The recombinant expressing system was confirmed by restriction endonuclease digestion, DNA sequencing and protein expression. E. coli cells were lysed by sonication and detergent dissolution. After cell membrane was extracted, the inclusion bodies were denatured by 8 mol/L Urea and purified with metal chelate affinity chromatography on Ni-NTA agarose under denaturing condition. The purified 6His-HPT was characterized by SDS-PAGE, and used to immunize rabbit. The titer and specificity of antisera were detected by ELISA and Western blot respecitively. RESULTS: Analysis of DNA sequence and restricted enzymes showed that the sequence of PBV222-HPT plasmid was correct. The amount of recombinant HPT expressed in E. coli accounted for 30% of total cellular proteins. From 1 liter of fermentative bacteria about 22 milligrams of pure recombinant HPT was isolated with purity above 95%. The recombinant HPT protein could produce high titer antiserum in rabbits and show good immunity activity. Western blot showed specific binding reaction between the antiserum to the purified 6His-HPT protein and their expressed products (plants protein and bacterial protein). CONCLUSION: HPT protein can be expressed and purified from E. coli by a relatively simple method, which has high immunity activity. Publication Types: Research Support, Non-U.S. Gov't PMID: 12964788 [PubMed - indexed for MEDLINE] 1442: Lancet. 2003 Aug 30;362(9385):714. GM foods in new dispute. Bosch X. Publication Types: News PMID: 12962112 [PubMed - indexed for MEDLINE] 1443: Nature. 2003 Sep 4;425(6953):15. Comment on: Nature. 2003 Jul 31;424(6948):473. Breeding to tackle blight without copper or GM. Duncan JM. Publication Types: Comment Letter PMID: 12955114 [PubMed - indexed for MEDLINE] 1444: Physiol Behav. 2003 Sep;79(4-5):783-8. Flavor preferences conditioned in C57BL/6 mice by intragastric carbohydrate self-infusion. Sclafani A, Glendinning JI. Department of Psychology, Brooklyn College and the Graduate School, The City University of New York, 2900 Bedford Avenue, Brooklyn, NY 11210-2889, USA. Asclafani@gc.cuny.edu This study determined the feasibility of conditioning flavor preferences in mice by self-administered intragastric (IG) nutrient infusions. Male C57BL/6J mice were surgically fitted with an IG catheter that was attached by a tether system to an infusion pump. The mice were given ad-libitum access to chow and a flavored solution 23 h/day. Drinking was monitored with a computerized lickometer system that controlled the infusion pumps. In Experiment 1, drinking one flavored solution (CS+, e.g., grape-saccharin) was paired with matched infusions of 8% maltodextrin, whereas drinking another solution (CS-, e.g., cherry-saccharin) was matched with water infusions across 6 one-bottle training days. During training, the mice drank more CS+ than CS-; this was due to an increase in bout size but not bout frequency. In subsequent two-bottle choice tests, the mice strongly preferred (91%) the CS+ to the CS-. Experiment 2 obtained a significant but less robust (71%) CS+ preference in mice trained with unsweetened CS solutions. These data indicate that mice, like rats, acquire an increased acceptance and preference for flavors paired with the postingestive actions of nutrients. Our understanding of flavor-nutrient learning can be advanced by studying this process in selected mouse strains and genetically modified animals. Publication Types: Comparative Study Research Support, U.S. Gov't, P.H.S. PMID: 12954423 [PubMed - indexed for MEDLINE] 1445: Health Serv J. 2003 Jul 24;113(5865):21. Abide with us. Hunt P. PMID: 12953678 [PubMed - indexed for MEDLINE] 1446: J Agric Food Chem. 2003 Sep 10;51(19):5695-702. Enhancement of the primary flavor compound methional in potato by increasing the level of soluble methionine. Di R, Kim J, Martin MN, Leustek T, Jhoo J, Ho CT, Tumer NE. Biotechnology Center for the Agriculture and the Environment, Department of Plant Biology and Pathology, Rutgers University, Cook College, 59 Dudley Road, New Brunswick, NJ 08901-8520, USA. The primary flavor compound in potato, methional, is synthesized from methionine by the Strecker degradation reaction. A major problem associated with potato processing is the loss of methional. Methional or its precursor, methionine, is not added back during potato processing due to high costs of production. A novel approach to enhance the methional level in processed potato would be to increase the production of its precursor, soluble methionine (Met). Cystathionine gamma-synthase (CGS) is a key enzyme regulating methionine biosynthesis in plants. To increase the level of soluble methionine in potato, Arabidopsis thaliana CGS cDNA was introduced under transcriptional control of the cauliflower mosaic virus 35S promoter into Russet Burbank potato by Agrobacterium-mediated transformation. Ten different transgenic potato lines (CGS1-10) were analyzed. Immunoblot analysis demonstrated that Arabidopsis CGS is expressed in the leaves, tubers, and roots of transgenic potato plants. CGS enzymatic activity was higher in the leaves and roots of the transgenic potato lines compared to the wild-type potato. Methionine levels in the leaves, roots and tubers of transgenic potato lines were enhanced as high as 6-fold compared to those in wild type potato plants. The methional level in baked tubers of field-grown transgenic potato lines was increased between 2.4- and 4.4-fold in lines CGS1, CGS2, and CGS4. The increase observed in methional levels correlated with the soluble methionine level in the tubers from the same lines measured before processing. These results provide the first evidence that the methional level can be enhanced in processed potatoes by increasing the production of its precursor, methionine. Publication Types: Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. PMID: 12952421 [PubMed - indexed for MEDLINE] 1447: EMBO Rep. 2003 Sep;4(9):819. Blame games. Breithaupt H. Publication Types: Editorial PMID: 12949577 [PubMed - indexed for MEDLINE] 1448: Nat Biotechnol. 2003 Sep;21(9):1003-9. Are Bt crops safe? Mendelsohn M, Kough J, Vaituzis Z, Matthews K. Office of Pesticide Programs of the U.S. Environmental Protection Agency, USA. mendelsohn.mike@epa.gov Publication Types: Evaluation Studies PMID: 12949561 [PubMed - indexed for MEDLINE] 1449: Nat Biotechnol. 2003 Sep;21(9):976. Comment on: Nat Biotechnol. 2003 Aug;21(8):852-4. Nat Biotechnol. 2003 Jul;21(7):735-6. EU reflects European public opinion. Borch K, Lassen J, Jørgensen RB. Publication Types: Comment Letter PMID: 12949553 [PubMed - indexed for MEDLINE] 1450: Nat Biotechnol. 2003 Sep;21(9):975-6. Comment on: Nat Biotechnol. 2003 Jul;21(7):737-8. EU reflects European public opinion. Barnes MH. Publication Types: Comment Letter PMID: 12949552 [PubMed - indexed for MEDLINE] 1451: Nat Biotechnol. 2003 Sep;21(9):974-5; author reply 975. Comment on: Nat Biotechnol. 2003 Jun;21(6):598. Identity tags revisited. Marillonnet S, Klimyuk V, Gleba Y. Publication Types: Comment Letter PMID: 12949551 [PubMed - indexed for MEDLINE] 1452: Nat Biotechnol. 2003 Sep;21(9):971-2. Acceptance of golden rice in the Philippine 'rice bowl'. Chong M. Publication Types: Letter Research Support, Non-U.S. Gov't PMID: 12949549 [PubMed - indexed for MEDLINE] 1453: Nat Biotechnol. 2003 Sep;21(9):957. UK government caught in GM dilemma. Mitchell P. Publication Types: News PMID: 12949540 [PubMed - indexed for MEDLINE] 1454: Trends Biotechnol. 2003 Sep;21(9):389-93. Labeling to manage marketing of GM foods. Smyth S, Phillips PW. Agricultural Economics, University of Saskatchewan, 51 Campus Drive, Saskatoon, Saskatchewan, S7N 5A8, Canada. sjs064@mail.usask.ca Biotechnology has the potential to introduce new food safety risks, liabilities and benefits, and although privately managed supply chains (involving proactive management of the production of branded products) are effective at providing, managing and communicating adequate information about products with well understood risks, products with uncertain risks pose a greater challenge. The demand for increased product information regarding genetically modified content, in particular, places new constraints on food supply chains, frequently resulting in communication failures. Here we assess and reject mandatory labeling as an appropriate response. PMID: 12948671 [PubMed - indexed for MEDLINE] 1455: Nature. 2003 Aug 28;424(6952):995. Comment on: Nature. 2003 Jul 31;424(6948):473. Consumers don't want GM, so why use it? Melchett P. Publication Types: Comment Letter PMID: 12944939 [PubMed - indexed for MEDLINE] 1456: Plant J. 2003 Jan;33(1):19-46. The release of genetically modified crops into the environment. Part II. Overview of ecological risk assessment. Conner AJ, Glare TR, Nap JP. New Zealand Institute for Crop & Food Research Ltd, Private Bag 4704, Christchurch, New Zealand. connert@crop.cri.nz Despite numerous future promises, there is a multitude of concerns about the impact of GM crops on the environment. Key issues in the environmental assessment of GM crops are putative invasiveness, vertical or horizontal gene flow, other ecological impacts, effects on biodiversity and the impact of presence of GM material in other products. These are all highly interdisciplinary and complex issues. A crucial component for a proper assessment is defining the appropriate baseline for comparison and decision. For GM crops, the best and most appropriately defined reference point is the impact of plants developed by traditional breeding. The latter is an integral and accepted part of agriculture. In many instances, the putative impacts identified for GM crops are very similar to the impacts of new cultivars derived from traditional breeding. When assessing GM crops relative to existing cultivars, the increased knowledge base underpinning the development of GM crops will provide greater confidence in the assurances plant science can give on the risks of releasing such crops. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 12943539 [PubMed - indexed for MEDLINE] 1457: J Plant Physiol. 2003 Jul;160(7):821-9. Vitamin production in transgenic plants. Herbers K. SunGene GmbH & Co. KGaA, Corrensstr. 3, D-06466 Gatersleben, Germany. karin.herbers@sungene.de Plants are a major source of vitamins in the human diet. Due to their significance for human health and development, research has been initiated to understand the biosynthesis of vitamins in plants. The pathways that are furthest advanced in elucidation are those of provitamin A, vitamin C and vitamin E. There is little knowledge about the regulation, storage, sink and degradation of any vitamin made in plants, or the interaction of vitamin biosynthetic pathways with other metabolic pathways. Researchers as well as life science companies have endeavoured to manipulate levels of vitamins in order to create functional food with enhanced health benefits, and even with the goal of achieving levels worth extracting from plant tissues. Thus far, metabolic engineering has resulted in transgenic plants that contain elevated levels of provitamin A, vitamin C and E, respectively. Additional research is necessary to identify all relevant target genes in order to further improve and tailor plants with elevated vitamin contents at will. Publication Types: Review PMID: 12940549 [PubMed - indexed for MEDLINE] 1458: J Plant Physiol. 2003 Jul;160(7):811-20. Engineering fructan metabolism in plants. Ritsema T, Smeekens SC. Molecular Plant Physiology, Utrecht University, Padualaan 8, 3584 CH Utrecht, The Netherlands. t.ritsema@bio.uu.nl Fructans, or polyfructosylsucroses, are storage carbohydrates present in many higher plants. They are also considered healthy food ingredients. Engineering crops into high level production of specific fructan molecules is one of the mayor strategic research goals. Understanding the properties of fructosyltransferases is important, in order to direct the synthesis of fructans. In plants at least two fructosyltransferases are needed to synthesise fructans. One enzyme synthesises the fructan trisaccharide 1-kestose, the next enzyme uses 1-kestose for elongation and/or modification, producing longer fructans. The specificity of fructosyltransferases determines the type of glycosidic bond formed and the donor and acceptor substrates used. This enables the synthesis of many structurally diverse fructans. The production of these molecules in crops such as sugar beet and potato makes the commercial use of fructans feasible. Publication Types: Review PMID: 12940548 [PubMed - indexed for MEDLINE] 1459: J Plant Physiol. 2003 Jul;160(7):755-64. Benefits and risks of antibody and vaccine production in transgenic plants. Warzecha H, Mason HS. Boyce Thompson Institute for Plant Research, Tower Road, Ithaca, New York 14850, USA. Phytopharming, the production of protein biologicals in recombinant plant systems, has shown great promise in studies performed over the past 13 years. A secretory antibody purified from transgenic tobacco was tested successfully in humans, and prevented bacterial re-colonization after topical application in the mouth. Rapid production of patient-tailored anti-lymphoma antibodies in recombinant Tobamovirus-infected tobacco may provide effective cancer therapy. Many different candidate vaccines from bacterial and viral sources have been expressed in transgenic plants, and three human clinical trials with oral delivery of transgenic plant tissues have shown exciting results. The use of crop plants with agricultural practice could allow cheap production of valuable proteins, while providing enhanced safety by avoidance of animal viruses or other contaminants. However development of this technology must carefully consider the means to ensure the separation of food and medicinal products when crop plants are used for phytopharming. Publication Types: Review PMID: 12940544 [PubMed - indexed for MEDLINE] 1460: J Plant Physiol. 2003 Jul;160(7):735-42. Overview of the current status of genetically modified plants in Europe as compared to the USA. Brandt P. Robert Koch-Institut, Zentrum Gentechnologie,Wollankstrasse 15-17, D-13187 Berlin, Germany. brandtp@rki.de Genetically modified crops have been tested in 1,726 experimental releases in the EU member states and in 7,815 experimental releases in the USA. The global commercial cultivation area of genetically modified crops is likely to reach 50 million hectares in 2001, however, the commercial production of genetically modified crops in the EU amounts to only a few thousand hectares and accounts for only some 0.03% of the world production. A significant gap exists between the more than fifty genetically modified crop species already permitted to be cultivated and to be placed on the market in the USA, Canada and other countries and the five genetically modified crop species permitted for the same use in the EU member states, which are still pending inclusion in the Common Catalogue of agricultural plant species. The further development of the "green gene technology" in the EU will be a matter of public acceptance and administrative legislation. Publication Types: Comparative Study PMID: 12940542 [PubMed - indexed for MEDLINE] 1461: J Plant Physiol. 2003 Jul;160(7):727-34. Discussion of current status of commercialization of plant biotechnology in the global marketplace. Emrich R. BASF Plant Science Holding GmbH, Agricultural Center, Building Li 554, 67117 Limburgerhof, Germany. reiner.emrich@basf-ag.de PMID: 12940541 [PubMed - indexed for MEDLINE] 1462: Adv Biochem Eng Biotechnol. 2003;84:1-48. The way ahead--the new technology in an old society. Sharma M, Swarup R. Department of Biotechnology, Ministry of Science & Technology, Government of India, New Delhi, India. manju@dbt.nic.in Biotechnology is one of the most important scientific and technological revolutions of the last century and has greatly benefited various aspects of human life. The potentials are enormous and many breakthroughs have already been achieved in the area of healthcare, food, agricultural products and environmental production. The developments in this important area provide immediate benefits to mankind and offer environmentally friendly technologies for sustainable development. The Department of Biotechnology, Government of India, set up in 1986, has played an important catalytic role in promoting this revolutionary field. Research and development, technology validation and demonstration, technology transfer, human resource development, setting up of Centers of Excellence and promoting industry-academia interactions have been some of the major achievements during the last 15 years. A unique feature of this Department is the strong interaction with scientists and institutes across the country to promote biotechnology research and development efforts for commercialization and also to benefit the rural population for socio-economic development. A large number of research institutes/universities and organizations across the country have been supported in the areas of agriculture, healthcare, environment and industry. In addition, basic research has also been an important thrust area. In order to ensure that the benefits of biotechnology reach the masses at large, a very stringent biosafety mechanism has been adopted. India is a country rich in biodiversity with two hot spots and has a strong base of expertise available in nearly all fields--thus biotechnology could flourish leading to a Bioindustrial Revolution. We are today poised to be the leaders in this field in the 21st Century. Publication Types: Review PMID: 12934932 [PubMed - indexed for MEDLINE] 1463: Curr Biol. 2003 Aug 19;13(16):R625-6. GM tensions dig in. Williams N. Publication Types: News PMID: 12932332 [PubMed - indexed for MEDLINE] 1464: Mol Ecol. 2003 Sep;12(9):2439-46. Oryzacystatin I expressed in transgenic potato induces digestive compensation in an insect natural predator via its herbivorous prey feeding on the plant. Bouchard E, Cloutier C, Michaud D. Département de Biologie, Centre de recherche en horticulture, Université Laval, Cité Universitaire (Québec), Canada G1K 7P4. We observed recently that the rice cysteine proteinase inhibitor, oryzacystatin I (OCI) expressed in transgenic potato does not affect growth and development of the two-spotted stinkbug predator (Perillus bioculatus) via its herbivorous prey feeding on the plant. Here we monitored the inhibitory activity of recombinant OCI along this potato --> herbivore --> predator continuum, to determine if the absence of effect was associated with a digestive compensatory response of the predator following inhibition of its proteinases by the recombinant cystatin. After confirming that OCI is present in the plant, and ingested in an active form by potato beetle larvae, quantitative and electrophoretic assays allowed us to determine that the recombinant cystatin (representing about 0.8% of total soluble proteins in leaves) was entirely bound to a approximately 30-kDa target proteinase in the prey's midgut, forming a sodium dodecyl sulphate (SDS)-stable complex detected on immunoblots with an anti-OCI polyclonal antibody. Despite the apparent absence of free, residual OCI in the beetle's midgut, digestive protease activity in the predator, known to include OCI-sensitive activity, was altered negatively when the prey was fed the modified plant. This inhibitory process at the third trophic level was accompanied by a compensatory response in the predator, by which serine-type proteinases were synthesized de novo. Overall, our data suggest that the affinity between OCI and the predator's OCI-sensitive proteinases is: (i) as strong as (or stronger than) the affinity between OCI and the potato beetle 30-kDa-sensitive proteinase; and (ii) stronger than the affinity between these enzymes and the plant endogenous homologue of OCI, potato multicystatin, induced in the plant by potato beetle feeding. Our results also show that predatory organisms can adapt their digestive metabolism to the presence of plant antidigestive proteins ingested by their herbivorous preys. In a broader context, this study stresses the need to monitor the inhibitory effects of PI-expressing plants not only on the herbivorous insects targeted, but also on the organisms likely to consume these pests in the environment. Publication Types: Comparative Study Research Support, Non-U.S. Gov't PMID: 12919481 [PubMed - indexed for MEDLINE] 1465: Mol Ecol. 2003 Sep;12(9):2429-37. Molecular interactions between an insect predator and its herbivore prey on transgenic potato expressing a cysteine proteinase inhibitor from rice. Bouchard E, Michaud D, Cloutier C. Département de Biologie, Centre de recherche en horticulture, Université Laval, Cité Universitaire (Québec), Canada G1K 7P4. Transgenic plants expressing resistance to herbivorous insects may represent a safe and sustainable pest control alternative if they do not interfere with the natural enemies of target pests. Here we examined interactions between oryzacystatin I (OCI), a proteinase inhibitor from rice genetically engineered into potato (Solanum tuberosum cv. Kennebec, line K52) to increase resistance to insect herbivory, and the insect predator Perillus bioculatus. This stinkbug is a relatively specialized predator of caterpillars and leaf-beetle larvae, and may also include plant sap in its predominantly carnivorous diet. One of its preferred prey is Colorado potato beetle (Leptinotarsa decemlineata), a major target of insect resistance development for potato field crops. Gelatin/sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) confirmed that a major fraction of proteinase (gelatinase) activity in P. bioculatus extracts is OCI-sensitive. Among five gelatinolytic bands detected, the slowest-moving one (proteinase I) was inhibited strongly by purified OCI expressed in Escherichia coli or by OCI-transgenic potato extracts, while three other proteinases were partly sensitive to these treatments. There was also evidence of slight inhibition of proteinase I by untransformed potato foliage, suggesting the presence of a natural inhibitor related to OCI at low level in potato foliage. Interestingly, only about 50% of the maximum potential activity of proteinase I was recovered in extracts of P. bioculatus feeding on L. decemlineata larval prey on a diet of OCI-potato foliage, indicating that the predator was sensitive to OCI in the midgut of its prey. However, P. bioculatus on OCI-prey survived, grew and developed normally, indicating ability to compensate prey-mediated exposure to the OCI inhibitor. Confinement of P. bioculatus to potato foliage provided no evidence that potato plant-derived nutrition is a viable alternative to predation, restriction to potato foliage in fact being inferior to free water for short-term survival of nonfeeding first-instar larvae. These results support the view that OCI, an effective inhibitor of a substantial fraction of digestive enzymatic potential in P. bioculatus, should not interfere with its predation potential when expressed in potato plants fed to its prey at a maximum level of approximately 0.8% of total soluble proteins in mature foliage. Publication Types: Comparative Study Research Support, Non-U.S. Gov't PMID: 12919480 [PubMed - indexed for MEDLINE] 1466: Genet Mol Res. 2003 Mar 31;2(1):112-6. Challenges when transferring technology from Lactococcus laboratory strains to industrial strains. Johansen E. Applied Biotechnology, Chr Hansen A/S, 10-12 Bøge Allé, Hørsholm, Denmark. Eric.Johansen@dk.chr-hansen.com Many genetically modified Lactococcus strains have been constructed in research laboratories around the world. Most of these have originated from laboratory strains and therefore there are several barriers to using them in an industrial setting. Laboratory strains are often plasmid-free and consequently Lac- and Prt-, rendering them unable to grow in milk. Many of the commonly used techniques have been optimised for laboratory strains and their application to industrial strains may require a great deal of effort. Often genetically modified organisms produced in the laboratory do not fit the published definition of 'food-grade' (Johansen, 1999, Encyclopedia of Food Microbiology, Academic Press, London, pp. 917-921) and a great deal of effort is required to eliminate undesirable DNA sequences. As a consequence, it is often necessary to recreate the strains in industrial backgrounds before the innovations described in the scientific literature can be applied to the real-world dairy industry. PMID: 12917807 [PubMed - indexed for MEDLINE] 1467: J Nutr Biochem. 2003 Jul;14(7):386-93. Stability of recombinant human alpha-1-antitrypsin produced in rice in infant formula. Chowanadisai W, Huang J, Huang N, Lönnerdal B. Department of Nutrition, University of California, Davis 95616, USA. Human milk contains several biologically active proteins that benefit the breast-fed infant. In order to survive in the gastrointestinal tract, these proteins need to be protected against proteolysis. Since human milk contains relatively high concentrations of alpha-1-antitrypsin (AAT), we have expressed recombinant AAT in rice to explore the possibility of supplementing infant formula with this protein. The stability of recombinant AAT was examined by biochemical and functional assays, such as SDS-PAGE, Western blotting, ELISA, elastase and trypsin inhibition, following exposure to heat, low pH, and in vitro digestion, conducted in both phosphate buffered saline and infant formula. Native AAT is resistant to acidic environments down to pH 2 for 1 h and can withstand in vitro digestion modeled after conditions in the infant gut. Recombinant AAT is nearly as resistant as the native form in buffer, and is equally resilient in formula. Heat treatments (60 degrees C for 15 min, 72 degrees C for 20 sec, 85 degrees C for 3 min, and 137 degrees C for 20 sec) revealed that recombinant AAT is not as stable as native AAT in buffer, particularly at higher temperatures. While significantly less recombinant AAT is detected by ELISA after heating in formula, addition of bile extract can restore epitopes resulting in higher concentrations, suggesting protein aggregation that may not affect AAT activity. This study shows that recombinant AAT may survive the conditions of the infant stomach and duodenum and affect protein digestion in the infant small intestine. Publication Types: Comparative Study Research Support, Non-U.S. Gov't PMID: 12915219 [PubMed - indexed for MEDLINE] 1468: Wei Sheng Yan Jiu. 2003 May;32(3):239-45. [Progress on biosafety assessment of marker genes in genetically modified foods] [Article in Chinese] Yang L, Yang X. Institute of Nutrition and Food Safety, Chinese Center for Disease Control and Prevention, Beijing 100050, China. Marker genes are useful in facilitating the detection of genetically modified organisms(GMO). These genes play an important role during the early identification stage of GMO development, but they exist in the mature genetically modified crops. So the safety assessment of these genes could not be neglected. In this paper, all the study on the biosafety assessment of marker genes were reviewed, their possible hazards and risks were appraised, and the marker genes proved safe were list too. GMO Labeling the is one important regulations for the development of genetically modified foods in the market. The accurate detecting techniques for GMO are the basis for setting up labeling regulation. In addition, some methods used to remove marker genes in genetically modified foods were introduced in the paper, which can eliminate their biosafety concern thoroughly. Publication Types: English Abstract Research Support, Non-U.S. Gov't Review PMID: 12914289 [PubMed - indexed for MEDLINE] 1469: Plant Physiol. 2003 Aug;132(4):1770-4. Going to "great lengths" to prevent the escape of genes that produce specialty chemicals. Ellstrand NC. Department of Genetics and Biotechnology Impacts Center, University of California, Riverside, California 92521-0124, USA. ellstrand@ucrac1.ucr.edu Publication Types: Research Support, U.S. Gov't, Non-P.H.S. PMID: 12913134 [PubMed - indexed for MEDLINE] 1470: Food Chem Toxicol. 2003 Oct;41(10):1273-82. The use of consumption data to assess exposure to biotechnology-derived foods and the feasibility of identifying effects on human health through post-market monitoring. Hlywka JJ, Reid JE, Munro IC. Cantox Health Sciences International, Suite 308, 2233 Argentia Road, ON L5N 2X7 Mississauga, Canada. jhlywka@cantox.com The pre-market safety assessment of foods derived through biotechnology provides a scientific basis for concluding reasonable certainty of no harm and ensuring safety. At a minimum, the outcome of such an assessment provides sufficient information to estimate the likelihood of adverse effects on consumers, generally precluding the need for post-market monitoring. Post-market monitoring (PMM) may be appropriate under certain conditions where a better estimate of dietary exposure and/or nutritional consequence of a biotechnology-derived food is required, when a potential safety issue, such as allergenicity, cannot be adequately addressed through pre-market studies, or to corroborate dietary intakes of a nutritionally improved food with beneficial effects on human health. Monitoring programs must be hypothesis-driven, and are dependent upon the availability of accurate consumption data. Exposure assessment methods include both deterministic and probabilistic estimates of intakes using food supply data, individual dietary surveys, household surveys, or total diet studies. In the development of a monitoring approach, resource allocation should be dependent upon both the desired level of conservatism and the endpoint of interest. However, the cost of monitoring varies substantially, and the potential to determine causation may be limited. Publication Types: Review PMID: 12909259 [PubMed - indexed for MEDLINE] 1471: Nutr Rev. 2003 Jun;61(6 Pt 2):S135-40. Safety assessment of foods produced through agricultural biotechnology. Taylor SL. University of Nebraska, Lincoln, 143 Food Industry Complex, Lincoln, NE 68583-0919, USA. Often the main criticism of foods derived from biotechnology is concerns about food safety. Whereas most present-day biotechnology-derived foods are approximately 99% similar to their non-biotechnology counterparts, the scientific community must ensure the safety of the novel aspects of these foods. The three phases of safety assessment are discussed and the concept of substantial equivalence is explained. PMID: 12908745 [PubMed - indexed for MEDLINE] 1472: Nutr Rev. 2003 Jun;61(6 Pt 2):S124-34. An historical perspective from the Green Revolution to the gene revolution. Davies WP. Royal Agricultural College, Cirencester, Gloucestershire, United Kingdom, GL7 6JS. Since the 1960s conventional crop breeding has increased food production commesurate with the growing population. For agricultural development to continue, the exploitation of greater genetic diversity and modern biotechnology are becoming increasingly important. This article reviews the milestones achieved by the Green Revolution and many of the recent breakthroughs of modern biotechnology. Publication Types: Historical Article Review PMID: 12908744 [PubMed - indexed for MEDLINE] 1473: Nutr Rev. 2003 Jun;61(6 Pt 2):S117-23. A perspective from the food industry. Mansour M. Keller and Heckman, 1001 G Street NW, Suite 500 West, Washington, DC 20001, USA. Biotechnology has the potential to meet the ever-increasing demand for enhanced food production. However, several factors besides scientific and agricultural advances can disrupt the progress of this field. This article reviews the interactions between political, regulatory, international, activist, consumer, and scientific entities. A discussion of the precautionary principle is included. Publication Types: Review PMID: 12908743 [PubMed - indexed for MEDLINE] 1474: Nutr Rev. 2003 Jun;61(6 Pt 2):S114-6. Productivity improvements in rice. Khush G. Department of Vegetable Crops, University of California, Davis, CA 95616, USA. Rice comprises 23%--an immense proportion-of the calories consumed worldwide. Countries that rely on rice as the main staple often consume up to 60% of their daily calories from this cereal product. During the Green Revolution, the yield potential of rice was greatly improved, which is especially important owing to the rapidly growing population. This article reviews the potential contributions of biotechnology to further rice yield potential. Publication Types: Review PMID: 12908742 [PubMed - indexed for MEDLINE] 1475: Nutr Rev. 2003 Jun;61(6 Pt 2):S105-9. Biotechnology, the environment, and sustainability. Kuiper H. Rikilt-Wageningen-UR, Postbus 230, 6700 AE, Wageningen, Netherlands. This article reviews the environmental impact of genetically modified crops through discussion of environmental risk assessment and includes several case studies. Influence of European regulatory and political issues is considered. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 12908740 [PubMed - indexed for MEDLINE] 1476: Nutr Rev. 2003 Jun;61(6 Pt 2):S101-4. Nutritionally enhanced rice to combat malnutrition disorders of the poor. Potrykus I. Swiss Federal Institute of Technology (ETH), Zurich, Switzerland. Major deficiency disorders, including vitamin A deficiency, are especially common in countries in which rice is the staple food. In response to the devastating effects of vitamin A deficiency, which may include blindness and, even death, "Golden Rice" has been developed to deliver this nutrient to those populations who need it most. The case of Golden Rice is used to demonstrate the challenges of radical GMO opposition, consumer acceptance, and regulation of biotechnology-derived foods. Publication Types: Review PMID: 12908739 [PubMed - indexed for MEDLINE] 1477: Curr Biol. 2003 Aug 5;13(15):R578-9. GM crops dancing to different tunes. Dixon B. MEDIAWATCH: Parts of the British media are keeping up a campaign against the introduction of genetically modified crops ahead of the governemnet's assessment of the trial programme this autumn, often to the detriment of the arguments. Publication Types: News PMID: 12906800 [PubMed - indexed for MEDLINE] 1478: Anal Bioanal Chem. 2003 Oct;377(3):496-506. Epub 2003 Aug 6. Biosensors based on enzyme field-effect transistors for determination of some substrates and inhibitors. Dzyadevych SV, Soldatkin AP, Korpan YI, Arkhypova VN, El'skaya AV, Chovelon JM, Martelet C, Jaffrezic-Renault N. Laboratory of Biomolecular Electronics, Institute of Molecular Biology & Genetics, National Academy of Sciences of Ukraine, 150 Zabolotnogo Str., 03143, Kiev, Ukraine. dzyad@yahoo.com This paper is a review of the authors' publications concerning the development of biosensors based on enzyme field-effect transistors (ENFETs) for direct substrates or inhibitors analysis. Such biosensors were designed by using immobilised enzymes and ion-selective field-effect transistors (ISFETs). Highly specific, sensitive, simple, fast and cheap determination of different substances renders them as promising tools in medicine, biotechnology, environmental control, agriculture and the food industry.The biosensors based on ENFETs and direct enzyme analysis for determination of concentrations of different substrates (glucose, urea, penicillin, formaldehyde, creatinine, etc.) have been developed and their laboratory prototypes were fabricated. Improvement of the analytical characteristics of such biosensors may be achieved by using a differential mode of measurement, working solutions with different buffer concentrations and specific agents, negatively or positively charged additional membranes, or genetically modified enzymes. These approaches allow one to decrease the effect of the buffer capacity influence on the sensor response in an aim to increase the sensitivity of the biosensors and to extend their dynamic ranges.Biosensors for the determination of concentrations of different toxic substances (organophosphorous pesticides, heavy metal ions, hypochlorite, glycoalkaloids, etc.) were designed on the basis of reversible and/or irreversible enzyme inhibition effect(s). The conception of an enzymatic multibiosensor for the determination of different toxic substances based on the enzyme inhibition effect is also described.We will discuss the respective advantages and disadvantages of biosensors based on the ENFETs developed and also demonstrate their practical application. Publication Types: Review PMID: 12904953 [PubMed - indexed for MEDLINE] 1479: Nature. 2003 Aug 7;424(6949):619. Genetic engineering: unnatural selection. Snow A. Department of Evolution, Ecology and Organismal Biology, Ohio State University, Columbus, Ohio 43210, USA. PMID: 12904767 [PubMed - indexed for MEDLINE] 1480: Nature. 2003 Aug 7;424(6949):613. Flawed science underlies laws on transgenic crops. Sandermann H Jr. Publication Types: Letter PMID: 12904760 [PubMed - indexed for MEDLINE] 1481: J Immunol. 2003 Aug 15;171(4):2116-26. A plant-based allergy vaccine suppresses experimental asthma via an IFN-gamma and CD4+CD45RBlow T cell-dependent mechanism. Smart V, Foster PS, Rothenberg ME, Higgins TJ, Hogan SP. Allergy and Inflammation Research Group, Division of Molecular Bioscience, The John Curtin School of Medical Research, Australian National University, Canberra, ACT, Australia. Allergic asthma is currently considered a chronic airway inflammatory disorder associated with the presence of activated CD4(+) Th2-type lymphocytes, eosinophils, and mast cells. Interestingly, therapeutic strategies based on immune deviation and suppression have been shown to successfully attenuate the development of the asthma phenotype. In this investigation, we have for the first time used a genetically modified (GM) plant, narrow leaf lupin (Lupinus angustifolius L.), expressing a gene for a potential allergen (sunflower seed albumin) (SSA-lupin) to examine whether a GM plant/food-based vaccine strategy can be used to suppress the development of experimental asthma. We show that oral consumption of SSA-lupin promoted the induction of an Ag-specific IgG2a Ab response. Furthermore, we demonstrate that the plant-based vaccine attenuated the induction of delayed-type hypersensitivity responses and pathological features of experimental asthma (mucus hypersecretion, eosinophilic inflammation, and enhanced bronchial reactivity (airways hyperreactivity). The suppression of experimental asthma by SSA-lupin was associated with the production of CD4(+) T cell-derived IFN-gamma and IL-10. Furthermore, we show that the specific inhibition of experimental asthma was mediated via CD4(+)CD45RB(low) regulatory T cells and IFN-gamma. Thus, our data demonstrate that a GM plant-based vaccine can promote a protective immune response and attenuate experimental asthma, suggesting that plant-based vaccines may be potentially therapeutic for the protection against allergic diseases. PMID: 12902518 [PubMed - indexed for MEDLINE] 1482: Trends Parasitol. 2003 Aug;19(8):349-55. The first releases of transgenic mosquitoes: an argument for the sterile insect technique. Benedict MQ, Robinson AS. Entomology Unit, Food and Agriculture Organization/International Atomic Energy Agency Agriculture and Biotechnology Laboratory, Agency's Laboratories Seibersdorf, A-1400 Vienna, Austria. m.benedict@iaea.org Potential applications for reducing transmission of mosquito-borne diseases by releasing genetically modified mosquitoes have been proposed, and mosquitoes are being created with such an application in mind in several laboratories. The use of the sterile insect technique (SIT) provides a safe programme in which production, release and mating competitiveness questions related to mass-reared genetically modified mosquitoes could be answered. It also provides a reversible effect that would be difficult to accomplish with gene introgression approaches. Could new technologies, including recombinant DNA techniques, have improved the success of previous mosquito releases? Criteria for an acceptable transgenic sterile mosquito are described, and the characteristics of radiation-induced sterility are compared with that of current transgenic approaches. We argue that SIT using transgenic material would provide an essentially safe and efficacious foundation for other possible approaches that are more ambitious. Publication Types: Review PMID: 12901936 [PubMed - indexed for MEDLINE] 1483: J Biochem Mol Toxicol. 2003;17(4):223-34. Argemone oil induced cellular damage in the reproductive tissues of transgenic Drosophila melanogaster: protective role of 70 kDa heat shock protein. Mukhopadhyay I, Saxena DK, Bajpai VK, Chowdhuri DK. Embryotoxicology Section, Industrial Toxicology Research Centre, Lucknow 226001, Uttar Pradesh, India. We explored the reproductive toxicity of argemone oil and its principal alkaloid fraction in transgenic Drosophila melanogaster (hsp70-lacZ) Bg(9). The toxicity of argemone oil has been attributed to two of its physiologically active benzophenanthridine alkaloids, sanguinarine and dihydrosanguinarine. Freshly eclosed first instar larvae of transgenic Drosophila melanogaster were transferred to different concentrations of argemone oil and its alkaloid fraction contaminated food. Virgin flies that eclosed from the contaminated food were pair-mated to look into the effect on reproduction. The study was further extended by investigating hsp70 expression and tissue damage in larval gonads, genital discs, and reproductive organs of adult fly. Our results showed that argemone oil was more cytotoxic than its principal alkaloid fraction. Moreover, it was the male fly that was more affected compared to its opposite number. The accessory glands of male reproductive system of the fly, which did not express hsp70, exhibited severe damage as evidenced by Trypan blue staining. This prompted us to explore the ultrastructural morphology of the gland, which showed acute signs of necrosis in both the cell types as evident by necrotic nuclei, higher vacuolization, and disorganized endoplasmic reticulum, decrease in the number of Golgi vesicles and disorganized, loosely packed filamentous structures in the lumen of the accessory gland, at the higher concentrations of the adulterant. The study showed the reproductive toxicity of argemone oil and its alkaloid fraction in transgenic Drosophila melanogaster and further confirmed the cytoprotective role of hsp70. Copyright 2003 Wiley Periodicals, Inc. J Biochem Mol Toxicol 17:223-234, 2003; Published online in Wiley InterScience (www.interscience.wiley.com). DOI 10.1002/jbt.10082. Publication Types: Research Support, Non-U.S. Gov't PMID: 12898646 [PubMed - indexed for MEDLINE] 1484: Nat Biotechnol. 2003 Aug;21(8):852-4. Comment in: Nat Biotechnol. 2003 Sep;21(9):976. Rethinking US leadership in food biotechnology. Taylor MR. Resources for the Future, 1616 P Street, NW Washington, DC 20036, USA. taylor@rff.org PMID: 12894194 [PubMed - indexed for MEDLINE] 1485: Nat Biotechnol. 2003 Aug;21(8):849-51. The science and politics of plant biotechnology--a personal perspective. Vasil IK. University of Florida, Box 110690, Gainesville, Florida 32611-0690, USA. ikv@mail.ifas.ufl.edu PMID: 12894193 [PubMed - indexed for MEDLINE] 1486: Nat Biotechnol. 2003 Aug;21(8):835-6. Comment in: Nat Biotechnol. 2004 Feb;22(2):149. GM labeling in China beset by problems. Jia H. Publication Types: News PMID: 12894183 [PubMed - indexed for MEDLINE] 1487: Nature. 2003 Jul 31;424(6948):473. Comment in: Nature. 2003 Aug 28;424(6952):995. Nature. 2003 Sep 4;425(6953):15. Diversity in food technology. [No authors listed] Publication Types: Editorial PMID: 12891314 [PubMed - indexed for MEDLINE] 1488: Lancet. 2003 Jul 19;362(9379):219. Codex adopts new standards on GM foods, irradiation, and animal feed. Kapp C. Publication Types: News PMID: 12885509 [PubMed - indexed for MEDLINE] 1489: Genes Brain Behav. 2003 Feb;2(1):32-9. Is the food-entrainable circadian oscillator in the digestive system? Davidson AJ, Poole AS, Yamazaki S, Menaker M. Collaborators: Menaker M. Department of Biology, University of Virginia, Charlottesville, Virginia 22904, USA. Food-anticipatory activity (FAA) is the increase in locomotion and core body temperature that precedes a daily scheduled meal. It is driven by a circadian oscillator but is independent of the suprachiasmatic nuclei. Recent results that reveal meal-entrained clock gene expression in rat and mouse peripheral organs raise the intriguing possibility that the digestive system is the site of the feeding-entrained oscillator (FEO) that underlies FAA. We tested this possibility by comparing FAA and Per1 rhythmicity in the digestive system of the Per1-luciferase transgenic rat. First, rats were entrained to daytime restricted feeding (RF, 10 days), then fed ad libitum (AL, 10 days), then food deprived (FD, 2 days). As expected FAA was evident during RF and disappeared during subsequent AL feeding, but returned at the correct phase during deprivation. The phase of Per1 in liver, stomach and colon shifted from a nocturnal to a diurnal peak during RF, but shifted back to nocturnal phase during the subsequent AL and remained nocturnal during food deprivation periods. Second, rats were entrained to two daily meals at zeitgeber time (ZT) 0400 and ZT 1600. FAA to both meals emerged after about 10days of dual RF. However, all tissues studied (all five liver lobes, esophagus, antral stomach, body of stomach, colon) showed entrainment consistent with only the night-time meal. These two results are inconsistent with the hypothesis that FAA arises as an output of rhythms in the gastrointestinal (GI) system. The results also highlight an interesting diversity among peripheral oscillators in their ability to entrain to meals and the direction of the phase shift after RF ends. Publication Types: Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. PMID: 12882317 [PubMed - indexed for MEDLINE] 1490: Science. 2003 Jul 25;301(5632):447-8. Agbiotech. U.K. government panel gives GM crops cautious support. Pickrell J. Publication Types: News PMID: 12881538 [PubMed - indexed for MEDLINE] 1491: Regul Toxicol Pharmacol. 2003 Aug;38(1):98-104. Post-market surveillance of GM foods: applicability and limitations of schemes used with pharmaceuticals and some non-GM novel foods. Wal JM, Hepburn PA, Lea LJ, Crevel RW. Laboratoire d' Immuno-Allergie Alimentaire, Service de Pharmacologie et d'Immunologie, INRA-CEA de Saclay, 91191 Gif sur Yvette Cedex, France. Post-market surveillance (PMS) is increasingly required by some regulatory authorities for the marketing approval of GM-Novel Foods. This requirement, in addition to a complete conventional safety assessment, aims to show that unexpected (adverse) effects do not occur after long-term everyday exposure. Large food manufacturers have systems to obtain feedback from consumers on their products. We show that such systems can be enhanced to collect information on possible health effects of specific products and relate these to intake in specific groups of consumers. The term post-launch monitoring (PLM) is proposed to distinguish the process from that used for pharmaceuticals. GM foods differ from branded products to which existing systems have been applied. The paper discusses whether and how such systems could be applied to GM foods and what additional elements would need to be incorporated in them. A PLM system should define and organize the flow of information between the different stakeholders. We conclude that because such data will be generated from a range of sources and will need to be collated, verified, and integrated, an independent agency will be essential to undertake this activity in order to balance the interests of all stakeholders and ensure public trust. Publication Types: Review PMID: 12878059 [PubMed - indexed for MEDLINE] 1492: Regul Toxicol Pharmacol. 2003 Aug;38(1):27-35. On the safety of a new generation of DSM Aspergillus niger enzyme production strains. van Dijck PW, Selten GC, Hempenius RA. DSM Food Specialties, Department of Regulatory Affairs, P.O. Box 1, 2600 MA Delft, Netherlands. piet.dijck-van@dsm.corp Consumers safety of enzyme preparations is determined by three variables: the producing organism, the raw materials used in the production, and the production process itself. The latter one is embedded in current Good Manufacturing Practice (cGMP) and Hazard Analysis of Critical Control Points (HACCP); therefore the safety focus can be directed to raw materials and the producing organism. In this paper, we describe the use of novel genetically modified strains of Aspergillus niger-made by a design and build strategy-from a lineage of classically improved strains with a history of safe use in enzyme production. The specifics of the host strain allow for integration and over-expression of any gene of interest at a targeted integration site implying that the rest of the host genome is not affected by this integration. Furthermore due to the fact that the newly integrated gene copies are put under the genetic regulation of the host's own glucoamylase promoter, the recipe of the production process of any new production strain can be kept constant with respect to the raw materials composition. Consequently the safety of a new enzyme product from these novel genetically modified strains is determined by the background of the production organism. The use of a strain with a history of safe use and targeted integration according to the concept described above has consequences for the safety studies on the final product. If a known enzymatic activity is over-expressed the safety of a new enzyme preparation is covered by the results of the safety studies performed for other strains from this specific Aspergillus niger strain lineage. In this paper an overview is given on the available toxicity tests with these strains. We conclude that for new enzyme products produced with strains from this lineage using the design and build technology no new sub-acute/chronic oral toxicity studies are needed. This also has the benefit that no longer test animals are needed to demonstrate the safety of products produced by these strains. Publication Types: Review PMID: 12878051 [PubMed - indexed for MEDLINE] 1493: Vopr Pitan. 2003;72(3):20-3. [Monitoring of food products from genetically modified sources in Moscow] [Article in Russian] Tutel'ian VA, Filatov NN, Sorokina EIu, Chernysheva ON, Salova NIa, Sizykh EV, Anisimova OV. This paper presents results of a detection of genetically modified organisms (GMO) in food from the shops of Moscow. The screening methods and event-specific assay based on the polymerase chain reaction is used. Transgenic DNA from genetically modified soybeans line 40-3-2 is detected in 17.2% samples of studied foods. Soybeans line 40-3-2 is allowed in Russian food supply. Publication Types: English Abstract PMID: 12872657 [PubMed - indexed for MEDLINE] 1494: Lancet. 2003 Jul 12;362(9378):135. Europe imposes strict GM-food laws. Bosch X. Publication Types: News PMID: 12870479 [PubMed - indexed for MEDLINE] 1495: Nature. 2003 Jul 17;424(6946):237. Comment in: Nature. 2003 Sep 25;425(6956):343. Don't believe the hype. [No authors listed] Publication Types: Editorial PMID: 12867939 [PubMed - indexed for MEDLINE] 1496: J Nutr Educ Behav. 2003 Jul-Aug;35(4):210-4. The developing world benefits from plant biotechnology. Mackey MA. Monsanto Company, Chesterfield, Missouri, USA. maureen.a.mackey@monsanto.com This article presents the view that the developing world benefits from plant biotechnology. To support this view, evidence is presented showing that plant biotechnology has already had a beneficial impact on millions of farmers of the developing world who have chosen to grow Bt cotton, resulting in fewer pesticide sprayings and increased yields and profits. Leaders, scientists, and farmers in developing countries have expressed their need for plant biotechnology, and in some developing countries, ongoing projects are applying plant biotechnology to improve indigenous crops. Nutrition education efforts should evaluate crop biotechnology from the perspective of the developing world. PMID: 12859886 [PubMed - indexed for MEDLINE] 1497: Plant Physiol. 2003 Jul;132(3):1448-63. Expression of anthocyanins and proanthocyanidins after transformation of alfalfa with maize Lc. Ray H, Yu M, Auser P, Blahut-Beatty L, McKersie B, Bowley S, Westcott N, Coulman B, Lloyd A, Gruber MY. Saskatoon Research Centre, Agriculture and Agri-Food Canada, 107 Science Place, Saskatoon, Saskatchewan, Canada S7N 0X2. Three anthocyanin regulatory genes of maize (Zea mays; Lc, B-Peru, and C1) were introduced into alfalfa (Medicago sativa) in a strategy designed to stimulate the flavonoid pathway and alter the composition of flavonoids produced in forage. Lc constructs included a full-length gene and a gene with a shortened 5'-untranslated region. Lc RNA was strongly expressed in Lc transgenic alfalfa foliage, but accumulation of red-purple anthocyanin was observed only under conditions of high light intensity or low temperature. These stress conditions induced chalcone synthase and flavanone 3-hydroxylase expression in Lc transgenic alfalfa foliage compared with non-transformed plants. Genotypes containing the Lc transgene construct with a full-length 5'-untranslated region responded more quickly to stress conditions and with a more extreme phenotype. High-performance liquid chromatography analysis of field-grown tissue indicated that flavone content was reduced in forage of the Lc transgenic plants. Leucocyanidin reductase, the enzyme that controls entry of metabolites into the proanthocyanidin pathway, was activated both in foliage and in developing seeds of the Lc transgenic alfalfa genotypes. Proanthocyanidin polymer was accumulated in the forage, but (+)-catechin monomers were not detected. B-Peru transgenic and C1 transgenic populations displayed no visible phenotypic changes, although these transgenes were expressed at detectable levels. These results support the emerging picture of Lc transgene-specific patterns of expression in different recipient species. These results demonstrate that proanthocyanidin biosynthesis can be stimulated in alfalfa forage using an myc-like transgene, and they pave the way for the development of high quality, bloat-safe cultivars with ruminal protein bypass. Publication Types: Research Support, Non-U.S. Gov't PMID: 12857826 [PubMed - indexed for MEDLINE] 1498: Science. 2003 Jul 11;301(5630):167; author reply 167. Comment on: Science. 2003 Apr 4;300(5616):61-2. Democratization is more than lower prices. Cayford J. Publication Types: Comment Letter PMID: 12855789 [PubMed - indexed for MEDLINE] 1499: Nature. 2003 Jul 10;424(6945):116. Europe finds transgenic food hard to swallow. Giles J. Publication Types: News PMID: 12853914 [PubMed - indexed for MEDLINE] 1500: J Econ Entomol. 2003 Jun;96(3):931-4. Effect of MON810 Bt field corn on Helicoverpa zea (Lepidoptera: Noctuidae) cannibalism and its implications to resistance development. Horner TA, Dively GP. USDA, APHIS, PPQ, 4700 River Road, Unit 133, Riverdale, MD 20737, USA. Pairs of Helicoverpa zea (Boddie) larvae reared on diet-incorporated MON810 transgenic leaf tissue of field corn (Zea mays L.) were observed in the laboratory to characterize effects of sublethal levels of Bacillus thuringiensis variety kurstaki (Bt) Cry1Ab endotoxins on cannibalistic behavior and mortality. Feeding on sublethal levels of Bt corn reduced the frequency of cannibalistic behaviors exhibited by H. zea when uneven instars were paired together. Exposure to the Bt endotoxin had no significant effect on when cannibalistic mortality occurred or the level of mortality as a result of cannibalism. Assuming that H. zea larvae reared on nonBt corn tissue behaved in a similar way that resistant larvae would if feeding on Bt tissue, sublethal effects of Cry1Ab intoxication may reduce the chances of successful cannibalism by susceptible larvae and thus play a disproportionate role in the survival of multiple ear infestations. Furthermore, cannibalistic encounters could result in partially resistant larvae feeding on nontoxic food, thus temporarily providing an escape from exposure to the Bt endotoxin. These behavior alterations could increase the selective differential between susceptible individuals and those carrying resistance genes. Publication Types: Research Support, Non-U.S. Gov't PMID: 12852638 [PubMed - indexed for MEDLINE] 1501: Riv Biol. 2003 Jan-Apr;96(1):31-54. Engineering crops, a deserving venture. Lanfranco L. Department of Plant Biology, University of Turin, Viale P.A. Mattioli 25, 10125 Turin, Italy. luisa.lanfranco@unito.it Plant transformation has had a deep impact on several aspects of basic and applied research. Genetic transformation has offered new opportunities compared to traditional breeding practises since it allows the integration into a host genome of specific sequences leading to a strong reduction of the casualness of gene transfer. One of the first target areas was plant protection against pests, pathogens and environmental stresses while the recent plant engineering programs are aimed at increasing food quality, in particular at increasing nutritional characteristics of food crops. Moreover, transgenic plants, tissue or cell cultures represent an attractive biological system for producing heterologous proteins since they offer economic and qualitative benefits. High yield production can be obtained and large-scale commercial production will take advantage of the existing infrastructure for crop cultivation, processing and storage. There are also qualitative benefits since protein synthesis secretion and post-translational modifications are similar in plants and animal cells. There are no human viral pathogens harboured by plants: thus, especially for pharmaceuticals, plants represent the safer production system. Plant transformation has become an essential instrument also for basic research, in particular for the functional characterisation of genes identified by sequencing of whole genomes. Large collections of insertion mutants have been obtained in the model plant Arabidopsis to provide a high level of genome saturation that means 95% chance of inactivating any gene at least once. To instil greater public confidence in modern plant biotechnology recent advances have already been made to overcome the potential risks for human health and environment. Publication Types: Review PMID: 12852173 [PubMed - indexed for MEDLINE] 1502: Neuron. 2003 Jul 3;39(1):147-61. Comment in: Neuron. 2003 Jul 3;39(1):6-8. Developmental control of foraging and social behavior by the Drosophila neuropeptide Y-like system. Wu Q, Wen T, Lee G, Park JH, Cai HN, Shen P. Department of Cellular Biology, University of Georgia, Athens, GA 3060, USA. Animals display stereotyped behavioral modifications during development, but little is known about how genes and neural circuits are regulated to turn on/off behaviors. Here we report that Drosophila neuropeptide F (dNPF), a human NPY homolog, coordinates larval behavioral changes during development. The brain expression of npf is high in larvae attracted to food, whereas its downregulation coincides with the onset of behaviors of older larvae, including food aversion, hypermobility, and cooperative burrowing. Loss of dNPF signaling in young transgenic larvae led to the premature display of behavioral phenotypes associated with older larvae. Conversely, dNPF overexpression in older larvae prolonged feeding, and suppressed hypermobility and cooperative burrowing behaviors. The dNPF system provides a new paradigm for studying the central control of cooperative behavior. Publication Types: Research Support, U.S. Gov't, P.H.S. PMID: 12848939 [PubMed - indexed for MEDLINE] 1503: Pest Manag Sci. 2003 Jun-Jul;59(6-7):770-6. United States Department of Agriculture-Agricultural Research Service research on managing insect resistance to insecticides. Elzen GW, Hardee DD. USDA, ARS, Kika de la Garza Subtropical Agricultural Research Center, Beneficial Insects Research Unit, Weslaco, TX 78596, USA. gelzen@weslaco.ars.usda.gov Insecticide resistance has developed within many classes of pesticide, and over 500 species of insects and mites are resistant to one or more insecticides. Insecticide resistance and the consequent losses of food and fiber caused by failure to control insect and mite pests causes economic losses of several billion dollars worldwide each year. It is the goal of insect resistance management (IRM) to preserve useful pesticides by slowing, preventing or reversing development of resistance in pests. Important aspects of this goal are understanding the development of resistance and monitoring to determine ways to prevent its development. We describe programs specific to missions of the US Department of Agriculture, Agricultural Research Service, which are designed to characterize insecticide resistance in insects and mites with the goal of managing pests in an ecologically acceptable manner. Resistance management of cotton, potatoes, vegetables, melons, ornamentals, greenhouse crops, corn, stored grains, livestock, honeybees and mites, as well as management of transgenic crops are evaluated. We conclude that IRM is a vital part of stewardship of any pest management product and must be a combined effort of manufacturers, growers, consultants, extension services and grower organizations, working closely with regulators, to achieve logistically and economically feasible systems that prolong the effectiveness of all pest-control products. Publication Types: Review PMID: 12846328 [PubMed - indexed for MEDLINE] 1504: Plant Cell Rep. 2003 Sep;22(2):117-21. Epub 2003 Jul 4. Quantitative GFP fluorescence as an indicator of recombinant protein synthesis in transgenic plants. Richards HA, Halfhill MD, Millwood RJ, Stewart CN Jr. Food Safety Center, University of Tennessee, Knoxville, TN 37996, USA. The utility of green fluorescent protein (GFP) for biological research is evident. A fluorescence-based method was developed to quantify GFP levels in transgenic plants and protein extracts. Fluorescence intensity was linear with increasing levels of GFP over a range that encompasses transgene expression in plants by the cauliflower mosaic virus 35S promoter. Standard curves were used to estimate GFP concentration in planta and in protein extracts. These values were consistent with ELISA measurements of GFP in protein extracts from transgenic plants, indicating that the technique is a reliable measure of recombinant GFP expression. The levels of in planta GFP expression in both homozygous and hemizygous plants was then estimated. Homozygous transgenic plants expressed twice the amount of GFP than hemizygous plants, suggesting additive transgene expression. This methodology may be useful to simplify the characterization of transgene expression in plants. Publication Types: Comparative Study Research Support, U.S. Gov't, Non-P.H.S. PMID: 12845471 [PubMed - indexed for MEDLINE] 1505: Biosci Biotechnol Biochem. 2003 Jun;67(6):1319-26. Cloning, sequencing, and heterologous expression of a cellobiohydrolase cDNA from the basidiomycete Corticium rolfsii. Yasokawa D, Shimizu T, Nakagawa R, Ikeda T, Nagashima K. Section of Biotechnology, Department of Applied Technology, Hokkaido Food Processing Research Center, 589-4 Bunkyodai Midorimachi, Ebetsu, Hokkaido 069-0836, Japan. duke@foodhokkaido.gr.jp From a Corticium rolfsii cDNA library, a clone homologous to other fungal cellobiohydrolase (CBH1) genes was isolated using the polymerase chain reaction. In the nucleotide sequence, one 1.6 kb long open reading frame coding for a polypeptide of 530 amino acid residues was detected which showed 64% identity with CBH1 of Phanerochaete chrysosporium. With expression of the 1.8 kb cDNA using the Aspergillus oryzae expression system, we detected microcrystalline cellulose (Avicel) hydrolyzing activity in the culture supernatant. The secreted protein, accompanied by the activity, was 89 kDa by SDS-polyacrylamide gel electrophoresis. Publication Types: Research Support, Non-U.S. Gov't PMID: 12843660 [PubMed - indexed for MEDLINE] 1506: Am Herit Invent Technol. 2003 Summer;19(1):16-25. The growth of genetically modified foods. Heppenheimer TA. Publication Types: Historical Article PMID: 12841195 [PubMed - indexed for MEDLINE] 1507: Ann Allergy Asthma Immunol. 2003 Jun;90(6 Suppl 3):90-8. Food biotechnology: is this good or bad? Implications to allergic diseases. Helm RM. Arkansas Children's Hospital Research Institute, Little Rock, Arkansas 72202-3591, USA. helmrick@uams.edu BACKGROUND: Food biotechnology represents advancement in the traditional interspecies and intergeneric breeding methods for improving food supplies worldwide. With respect to safety, foods developed through biotechnology techniques represent one of the most extensively reviewed agricultural advancements in history. OBJECTIVE: To review the relevant issues with respect to foods from genetically modified crops and allergenicity. DATA SOURCES: To impart this information, the author will rely upon his experiences with investigations into food allergy and food allergens, participation in various workshops designed to determine allergenicity of novel proteins introduced into the diet, web sites, issue papers, and articles relevant to the topic. RESULTS: Given that there are no validated methods or models to determine potential allergenicity of novel proteins, criteria have been established based upon characteristics of known food allergens. The combination of genetic and bioinformatics information available from known food allergens applied to foods developed from genetically modified crops to avoid the inadvertent introduction of allergens into foods should pose no significant allergenic concern to individuals with a genetic predisposition to food allergy. Education and sound scientific evaluation provided to the consumer should alleviate any fear of emotionalism as implied by "Frankenfoods." CONCLUSIONS: The estimation that more than two trillion transgenic plants have been grown in 1999 and 2000 alone, with no overt documented adverse food reactions being reported, indicates that genetic modification through biotechnology will not impose immediate significant risks as food allergen sources beyond that of our daily dietary intake of foods from crop plants. Publication Types: Review PMID: 12839121 [PubMed - indexed for MEDLINE] 1508: Trends Biotechnol. 2003 Jul;21(7):294-7. Naturalness and the genetic modification of animals. Verhoog H. Louis Bolk Institute, Hoofdstraat 24, NL-3872 LA Driebergen, The Netherlands. h.verhoog@louisbolk.nl In the past few years it has been recognised that so-called intrinsic concerns about genetic modification (GM) of plants and animals, for food in particular, have an important role in the public perception of GM. One of these concerns is the view that GM is 'unnatural'. This article gives an overview of the often conflicting views on the argument of unnaturalness in books and reports. The author gives a new direction to this discussion, by contrasting the common sense view of nature and animals, with the scientific concept of nature and what is natural. The view of nature and what is natural is always normative. This is illustrated by making explicit the concept of nature in organic farming, which explains why GM is rejected. PMID: 12837612 [PubMed - indexed for MEDLINE] 1509: EMBO Rep. 2003 Jul;4(7):647-9. Food fights. Moore A. PMID: 12835747 [PubMed - indexed for MEDLINE] 1510: Development. 2003 Aug;130(16):3795-805. zag-1, a Zn-finger homeodomain transcription factor controlling neuronal differentiation and axon outgrowth in C. elegans. Wacker I, Schwarz V, Hedgecock EM, Hutter H. Max Planck Institute for Medical Research, Jahnstrasse 29, 69120 Heidelberg, Germany. The nervous system consists of diverse subtypes of neurons, whose identities must be specified during development. One important aspect of the differentiation program of neurons is the expression of the appropriate set of genes controlling axon pathway selection. We have identified a novel Znfinger/homeodomain containing transcription factor, zag-1, required for particular aspects of axonal pathfinding. In zag-1 mutants, motorneuron commissures either branch prematurely or fail to branch at the correct point. Ventral cord interneurons show defects in the guidance towards the ventral cord and also in the ventral cord. Several neurons misexpress differentiation markers, including glutamate receptor subunits and chemosensory receptors. zag-1 is expressed transiently in embryonic and postembryonic neurons during differentiation as well as in some mesodermal tissues. Null mutants of zag-1 are unable to swallow food and die as L1 larvae with a starved appearance, indicating that zag-1 has an additional role in pharynx development. The vertebrate homolog, deltaEF1, is highly conserved and known to act as transcriptional repressor in various tissues. Our data indicate that zag-1 also acts as transcriptional repressor controlling important aspects of terminal differentiation of neurons. Publication Types: Research Support, Non-U.S. Gov't PMID: 12835395 [PubMed - indexed for MEDLINE] 1511: Nat Biotechnol. 2003 Jul;21(7):739-41. Codex guidelines for GM foods include the analysis of unintended effects. Haslberger AG. University of Vienna and the World Health Organization FOS Program for Food Safety, Geneva, Switzerland. haslbergera@who.int PMID: 12833088 [PubMed - indexed for MEDLINE] 1512: Nat Biotechnol. 2003 Jul;21(7):737-8. Comment in: Nat Biotechnol. 2003 Sep;21(9):975-6. First salvo in transatlantic food fight is far from last word. Miller HI. Hoover Institution, Stanford University, Stanford, CA 94305-6010, USA. miller@hoover.stanford.edu PMID: 12833087 [PubMed - indexed for MEDLINE] 1513: Nat Biotechnol. 2003 Jul;21(7):735-6. Comment in: Nat Biotechnol. 2003 Sep;21(9):976. The WTO complaint--why now? Kinderlerer J. Sheffield Institute of Biotechnological Law and Ethics, Department of Law, Conduit Road, University of Sheffield, Sheffield S10 1FL, UK. j.kinderlerer@sheffield.ac.uk PMID: 12833086 [PubMed - indexed for MEDLINE] 1514: Nat Biotechnol. 2003 Jul;21(7):720-1. US academia held accountable for GM products. Powell K. Publication Types: News PMID: 12833077 [PubMed - indexed for MEDLINE] 1515: Medsurg Nurs. 2002 Oct;11(5):242-6. Genetically modified foods: a taste of the future. Lessick M, Keithley J, Swanson B, Lemon B. University of Toledo, College of Health and Human Services, Toledo, OH, USA. Technologies for genetically modifying foods hold tremendous promise for meeting important public health challenges in this century. By keeping informed of the ongoing development of genetically modified foods, nurses can effectively educate patients about the benefits and risks of these foods and promote informed decision making. Publication Types: Review PMID: 12830747 [PubMed - indexed for MEDLINE] 1516: Nature. 2003 Jun 26;423(6943):903. A flight of fancy. [No authors listed] Publication Types: Editorial PMID: 12827151 [PubMed - indexed for MEDLINE] 1517: Environ Health Perspect. 2003 Jun;111(8):1140-1. Assessment of allergenic potential of genetically modified foods: an agenda for future research. Selgrade MK, Kimber I, Goldman L, Germolec DR. National Health and Environmental Effects Research Laboratory, Office of Research and Development, U.S. Environmental Protection Agency, Research Triangle Park, North Carolina, USA. selgrade.maryjane@epa.gov Speakers and participants in the workshop "Assessment of the Allergenic Potential of Genetically Modified Foods" met in breakout groups to discuss a number of issues including needs for future research. These groups agreed that research should progress quickly in the area of hazard identification and that a need exists for more basic research to understand the mechanisms underlying food allergy. A list of research needs was developed. PMID: 12826487 [PubMed - indexed for MEDLINE] 1518: Environ Health Perspect. 2003 Jun;111(8):1131-9. Key issues for the assessment of the allergenic potential of genetically modified foods: breakout group reports. Germolec DR, Kimber I, Goldman L, Selgrade M. Laboratory of Molecular Toxicology, National Institute of Environmental Health Sciences, Research Triangle Park, North Carolina, USA. germolec@niehs.nih.gov On the final afternoon of the workshop "Assessment of the Allergenic Potential of Genetically Modified Foods," held 10-12 December 2001 in Chapel Hill, North Carolina, USA, speakers and participants met in breakout groups to discuss specific questions in the areas of use of human clinical data, animal models to assess food allergy, biomarkers of exposure and effect, sensitive populations, dose-response assessment, and postmarket surveillance. Each group addressed general questions regarding allergenicity of genetically modified foods and specific questions for each subject area. This article is a brief summary of the discussions of each of the six breakout groups regarding our current state of knowledge and what information is needed to advance the field. Publication Types: Review PMID: 12826486 [PubMed - indexed for MEDLINE] 1519: Environ Health Perspect. 2003 Jun;111(8):1125-30. Assessment of protein allergenicity on the basis of immune reactivity: animal models. Kimber I, Dearman RJ, Penninks AH, Knippels LM, Buchanan RB, Hammerberg B, Jackson HA, Helm RM. Syngenta Central Toxicology Laboratory, Alderley Park, Macclesfield, Cheshire, United Kingdom. ian.kimber@syngenta.com Because of the public concern surrounding the issue of the safety of genetically modified organisms, it is critical to have appropriate methodologies to aid investigators in identifying potential hazards associated with consumption of foods produced with these materials. A recent panel of experts convened by the Food and Agriculture Organization and World Health Organization suggested there is scientific evidence that using data from animal studies will contribute important information regarding the allergenicity of foods derived from biotechnology. This view has given further impetus to the development of suitable animal models for allergenicity assessment. This article is a review of what has been achieved and what still has to be accomplished regarding several different animal models. Progress made in the design and evaluation of models in the rat, the mouse, the dog and in swine is reviewed and discussed. Publication Types: Review PMID: 12826485 [PubMed - indexed for MEDLINE] 1520: Environ Health Perspect. 2003 Jun;111(8):1122-4. Protein digestibility and relevance to allergenicity. Bannon G, Fu TJ, Kimber I, Hinton DM. Product Safety Center, Monsanto, St. Louis, Missouri, USA. In January 2001 a Joint Food and Agriculture Organization of the United Nations/World Health Organization Expert Consultation Committee on Allergenicity of Foods Derived from Biotechnology published a report outlining in detail an approach for assessing the allergenic potential of novel proteins. One component of this decision tree is a determination of whether the protein of interest is resistant to proteolytic digestion. Although these (Italic)in vitro(/Italic) methodologies have been useful, the correlation between resistance to proteolysis and allergenic activity is not absolute. Two views and highlights of supporting research regarding the relationship of resistance to digestion and allergenicity are presented in this article. Publication Types: Review PMID: 12826484 [PubMed - indexed for MEDLINE] 1521: Environ Health Perspect. 2003 Jun;111(8):1114-21. Clinical and laboratory investigation of allergy to genetically modified foods. Bernstein JA, Bernstein IL, Bucchini L, Goldman LR, Hamilton RG, Lehrer S, Rubin C, Sampson HA. Department of Medicine, University of Cincinnati, Cincinnati, Ohio, USA. Technology has improved the food supply since the first cultivation of crops. Genetic engineering facilitates the transfer of genes among organisms. Generally, only minute amounts of a specific protein need to be expressed to obtain the desired trait. Food allergy affects only individuals with an abnormal immunologic response to food--6% of children and 1.5-2% of adults in the United States. Not all diseases caused by food allergy are mediated by IgE. A number of expert committees have advised the U.S. government and international organizations on risk assessment for allergenicity of food proteins. These committees have created decision trees largely based on assessment of IgE-mediated food allergenicity. Difficulties include the limited availability of allergen-specific IgE antisera from allergic persons as validated source material, the utility of specific IgE assays, limited characterization of food proteins, cross-reactivity between food and other allergens, and modifications of food proteins by processing. StarLink was a corn variety modified to produce a (Italic)Bacillus thuringiensis(/Italic) (Bt) endotoxin, Cry9C. The Centers for Disease Control and Prevention investigated 51 reports of possible adverse reactions to corn that occurred after the announcement that StarLink, allowed for animal feed, was found in the human food supply. Allergic reactions were not confirmed, but tools for postmarket assessment were limited. Workers in agricultural and food preparation facilities have potential inhalation exposure to plant dusts and flours. In 1999, researchers found that migrant health workers can become sensitized to certain Bt spore extracts after exposure to Bt spraying. Publication Types: Review PMID: 12826483 [PubMed - indexed for MEDLINE] 1522: Environ Health Perspect. 2003 Jun;111(8):1110-3. Introduction: what are the issues in addressing the allergenic potential of genetically modified foods? Metcalfe DD. Laboratory of Allergic Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland, USA. dean_metcalfe@nih.gov There is growing concern among the general public and the scientific community regarding the potential toxicity of genetically modified organisms (GMOs). The use of biotechnology to enhance pest resistance or nutritional value has raised a number of fundamental questions including the consequences of insertion of reporter genes, the spread of resistance genes to surrounding plants, and the use of suicide genes to prohibit reuse of seed from engineered plants. Of particular interest is the ability of proteins from GMOs to elicit potentially harmful immunologic responses, including allergic hypersensitivity. The lack of information of the potential toxicity of these products suggests a need to identify the critical issues and research needs regarding these materials and to develop testing strategies to examine the allergenicity of these compounds. Publication Types: Review PMID: 12826482 [PubMed - indexed for MEDLINE] 1523: Crit Rev Food Sci Nutr. 2003;43(3):245-64. A review on biological control and metabolism of aflatoxin. Mishra HN, Das C. Post Harvest Technology Centre, Department of Agricultural and Food Engineering, Indian Institute of Technology, Kharagpur-721302, India. hnm@agfe.iitkgp.ernet.in The series of events that led to the discovery of aflatoxin as a potent carcinogen, its biosynthesis, mechanism of action, structure-function relationship provide interesting insight into the economical and technological factors involved in the development of an effective control measure for the toxin. Scientists all over the world are making continuous efforts to explore a generalized process of detoxification, which can bring down the toxin content in heterogenous commodities to a threshold level. In this article biological control methods with special emphasis on in vivo and in vitro enzymatic detoxification of aflatoxin have been reviewed. Future areas of research involving large-scale enzymatic detoxification and modified atmosphere storage are also discussed. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 12822672 [PubMed - indexed for MEDLINE] 1524: Plant Cell Rep. 2003 Aug;21(12):1207-10. Epub 2003 Jun 18. Influence of Agrobacterium tumefaciens strain on the production of transgenic peas ( Pisum sativum L.). Grant JE, Thomson LM, Pither-Joyce MD, Dale TM, Cooper PA. New Zealand Institute for Crop and Food Research, Private Bag 4704, Christchurch, New Zealand. grantj@crop.cri.nz We compared the efficiency of two Agrobacterium tumefaciens strains, AGL 1 and KYRT1, for producing transgenic pea plants. KYRT1 is a disarmed strain of Chry5 that has been shown to be highly tumourigenic on soybean. The efficacies of the strains were compared using cotyledon explants from three pea genotypes and two plasmids. The peas were sourced from field-grown plants over three Southern Hemisphere summer seasons. Overall, KYRT1 was found to be on average threefold more efficient than AGL 1 for producing transgenic plants. We suggest that KYRT1 is sensitive to cocultivation temperature as the expected increase in efficiency was not achieved at high laboratory temperatures. Publication Types: Evaluation Studies Research Support, Non-U.S. Gov't PMID: 12819922 [PubMed - indexed for MEDLINE] 1525: BMJ. 2003 Jun 21;326(7403):1351. Controversy grows over India's genetically modified potato. Mudur G. Publication Types: News PMID: 12816815 [PubMed - indexed for MEDLINE] 1526: Nature. 2003 Jun 19;423(6942):823. Erratum in: Nature. 2004 Jul 22;430(6998):417. Comment in: Nature. 2003 Aug 7;424(6949):613. Producing decaffeinated coffee plants. Ogita S, Uefuji H, Yamaguchi Y, Koizumi N, Sano H. Research and Education Centre for Genetic Information, Nara Institute of Science and Technology, Japan. PMID: 12815419 [PubMed - indexed for MEDLINE] 1527: Nature. 2003 Jun 19;423(6942):790. Traditional wheat breeder fights for funding. Dalton R. Publication Types: News PMID: 12815388 [PubMed - indexed for MEDLINE] 1528: Lancet. 2003 Jun 14;361(9374):2051. Comment in: Lancet. 2003 Sep 6;362(9386):835. UK ethicists say GM foods could help poor. Group back the controversial crops as government warily tests public opinion. Ashraf H. PMID: 12814720 [PubMed - indexed for MEDLINE] 1529: Plant Physiol. 2003 Jun;132(2):979-87. Physiological and molecular assessment of altered expression of Hsc70-1 in Arabidopsis. Evidence for pleiotropic consequences. Sung DY, Guy CL. Plant Molecular and Cellular Biology Program, Department of Environmental Horticulture, Institute of Food and Agricultural Sciences, University of Florida, Gainesville 32611-0670, USA. Hsp70s function as molecular chaperones. The protective chaperone activities of hsp70 help to confer tolerance to heat, glucose deprivation, and drought. Overexpression of hsp70s in many organisms correlates with enhanced thermotolerance, altered growth, and development. To better understand the roles of hsp70 proteins in Arabidopsis, the molecular and physiological consequences of altered expression of the major heat shock cognate, Hsc70-1, were analyzed. Extensive efforts to achieve underexpression of Hsc70-1 mRNA using a full-length antisense cDNA resulted in no viable transgenic plants, suggesting that reduced expression is lethal. Constitutive overexpression of Hsc70-1 also appeared to be deleterious to viability, growth, and development because fewer transformants were recovered, and most were dwarfed with altered root systems. Despite being dwarfed, the overexpression plants progressed normally through four selected developmental stages. Heat treatment revealed that Hsc70-1 overexpression plants were more tolerant to heat shock (44 degrees C for 10 min). The elevated basal levels of HSC70-1 in transgenic plants led to delayed heat shock response of several heat shock genes. The data in this study suggest that tight regulation of Hsc70-1 expression is critical for the viability of Arabidopsis and that the functions of HSC70-1 contribute to optimum growth, development, thermotolerance, and regulation of the heat shock response. Publication Types: Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. PMID: 12805626 [PubMed - indexed for MEDLINE] 1530: Plant Physiol. 2003 Jun;132(2):748-56. Epub 2003 Apr 17. Expression of antisense acyl carrier protein-4 reduces lipid content in Arabidopsis leaf tissue. Branen JK, Shintani DK, Engeseth NJ. Department of Food Science and Human Nutrition, University of Illinois, 259 Edward R. Madigan Library, 1201 West Gregory Drive, Urbana 61801, USA. Arabidopsis plants were transformed with acyl carrier protein (ACP)-4 in antisense conformation driven by the cauliflower mosaic virus 35S promoter. It was hypothesized that reduction of ACP4 in leaf tissue would result in a reduction in lipid biosynthesis and, in addition, affect fatty acid composition and leaf physiology. Several transgenic lines have been generated with reduced ACP4 protein in leaf tissue. Dramatic reductions in ACP4 resulted in a reduction of leaf lipid content (22%-60%) based on fresh leaf weight and a bleached appearance and reduced photosynthetic efficiency. In addition, a decrease in 16:3 as a percentage of the total fatty acid composition was noted. There were no changes in leaf lipid class distribution; however, there was a decrease in the relative amount of 16:3 in monogalactosyldiacylglycerol. These results suggest that ACP4 plays a major role in the biosynthesis of fatty acids for chloroplast membrane development. Alterations in the ACP isoform profile of Arabidopsis leaf also appear to alter the flow of fatty acids between the prokaryotic and eukaryotic pathways for assembly of galactolipids. However, it has not yet been determined if the changes in fatty acid composition are due to changes in the profile of ACP isoforms, or if they are actually a reaction to a reduction in fatty acid precursors. Publication Types: Research Support, U.S. Gov't, Non-P.H.S. PMID: 12805604 [PubMed - indexed for MEDLINE] 1531: Science. 2003 Jun 13;300(5626):1637-8. Agricultural biotech. U.K. probes public opposition to GM crops. Eastman Q. Publication Types: News PMID: 12805509 [PubMed - indexed for MEDLINE] 1532: Nutr Health. 2003;17(1):1-8. In vivo studies on possible health consequences of genetically modified food and feed--with particular regard to ingredients consisting of genetically modified plant materials. Pryme IF, Lembcke R. Department of Biochemistry and Molecular Biology, University of Bergen, Arstadveien 19, NO-5009 Bergen, Norway. ian.pryme@ibmb.uib.no This synopsis reviews published in vivo studies on possible health consequences of genetically modified food and feed where the ingredients in question have consisted of genetically modified plant materials. The following, however, have not been taken into consideration:--ingredients consisting of genetically modified microorganisms or parts of animals/fish--ingredients produced by/from genetically modified organisms but without any DNA present--studies on consequences for the environment or biodiversity--in vitro studies or computer simulations. According to a Norwegian report "Gen-mat" (NOU 2000:29), and a more recent search in Medline and Citations Index, to our knowledge a total of ten studies have been published on the health effects of GM-foods and feeds. In this minireview the data made available in these published studies is discussed. Publication Types: Review PMID: 12803276 [PubMed - indexed for MEDLINE] 1533: Nature. 2003 Jun 12;423(6941):672. Public input sought on transgenic farming. Tilstone C. Publication Types: News PMID: 12802294 [PubMed - indexed for MEDLINE] 1534: Nature. 2003 Jun 12;423(6941):669. Debate, what debate? [No authors listed] Publication Types: Editorial PMID: 12802292 [PubMed - indexed for MEDLINE] 1535: Time. 2003 May 26;161(21):56-7. Cures on the cob. Roosevelt M. Publication Types: News PMID: 12800326 [PubMed - indexed for MEDLINE] 1536: Appetite. 2003 Jun;40(3):299-307. Dimensions of novelty: a social representation approach to new foods. Bäckström A, Pirttilä-Backman AM, Tuorila H. Department of Food Technology, University of Helsinki, P.O. Box 27, Helsinki FIN-00014, Finland. anna.backstrom@helsinki.fi Social representations of new foods were examined with a total of 44 subjects in nine focus groups. Each group was homogenous, defined by age, gender and educational background. Halfway through the interview, commercial packages of functional, genetically modified, organic, nutritionally modified and ethnic foods were presented as visual stimuli for discussion. Thematic and content analyses of the interview data showed that five dichotomies characterized the social representation: trust/distrust, safe/unsafe, natural/artificial, pleasure/necessity, and past/present. Many metaphors were used, with functional products being associated metaphorically with, for example, medicine and genetically modified products being associated with death and terrorism. Chronological references focused on the development of cuisine. The perceived unsafety of new foods was an important argument for women but not for men. The difference between age groups was in relating the discussion to either present time (young subjects) or past time (older subjects). Level of education affected the content of argumentation. In the context of new foods, social representations are formed to cope with the feeling of strangeness evoked by the novelties. They also have a role in cultural acceptance of new products by making them familiar. Overall, the results reflect the development of a new common sense in which popularized scientific notions are anchored in the process of urbanization. PMID: 12798788 [PubMed - indexed for MEDLINE] 1537: Wei Sheng Yan Jiu. 2003 Mar;32(2):134-7. [Studies on protein-based identification method of genetically modified capsicum] [Article in Chinese] Liu J, Deng P, Fang S, Zhao J. Shenzhen Disease Prevention and Control Center, Shenzhen 518020, China. The detection system based on protein is a method to evaluate the safety of genetically modified foods (GMF). Using cecropin BD gene in capsicum, a detecting method was set up. It is a system of evaluating the real expressive condition and safety of the foreign target protein of GMF. In this studies, with the preformative technic method, a satisfactory results by making use of hemolymph of immunized pupae of Antheraea pernyi as standard experimental material was achieved, comparing with the realities of the goal protein expressive condition of cecropin D gene in capsicum. The detecting steps were as following: the goal protein from material was extracted roughly, then with CM-Sepharose-FF ion-exchange chromatography twice, the goal protein was purified moderately. The purified product was identified by detecting the anti-bacterial activity, electrophoresis, biological auto-photography of the goal protein and MADDI-TOF mass spectrum. The results showed that the expressive foreign target protein in transgenic capsicum was in accordance with standard protein in the physical and chemical property, anti-bacterial activity and molecular weight. It indicated that expression of the target gene in capsicum is real, it corresponded to expected value. The separation, purification and identification methods of cecropin D were established in the study. By means of the comparative experiments about anti-bacterial activity and molecular weight of anti-bacterial peptide(ABP) from GM-capsicum and hemolymph of immunized pupae of Antheraea pernyi, the identification method of target protein from GM-capsicum was set up. The method is easy to be operated, fast and feasible. Publication Types: English Abstract PMID: 12793005 [PubMed - indexed for MEDLINE] 1538: Nature. 2003 Jun 5;423(6940):574. Divisions sink US consensus effort on transgenic food. Knight J. Publication Types: News PMID: 12789300 [PubMed - indexed for MEDLINE] 1539: Trends Biotechnol. 2003 Jun;21(6):249-50. Improved evaluation of potential allergens in GM food. Jank B, Haslberger AG. Publication Types: Letter PMID: 12788543 [PubMed - indexed for MEDLINE] 1540: J Invertebr Pathol. 2003 Jun;83(2):113-7. Precautionary risk assessment of Bt maize: what uncertainties? Levidow L. Centre for Technology Strategy, Open University, Milton Keynes, MK7 6AA, UK. L.Levidow@open.ac.uk GM crops have become a test case for the conflicting slogans of 'the precautionary principle' versus 'sound science.' The issues can be illustrated by developments in regulatory science for Bt maize in the European Union. As this case study suggests, risk assessment is always framed by some account of the relevant uncertainties. These in turn depend upon how the environment is valued and how scientific questions are posed about cause-effect pathways of potential harm. The slogan of 'sound science' hides such judgements, by representing ignorance or value-judgements as 'science.' By contrast, precaution can challenge such judgements, identify new unknowns, generate different criteria for evidence, open up new scientific questions, and make these judgements more transparent. It is doubtful whether these complexities have been fully acknowledged by specialists, and thus whether the continued risk debate is due solely to a public misunderstanding of science. Publication Types: Review PMID: 12788280 [PubMed - indexed for MEDLINE] 1541: J Invertebr Pathol. 2003 Jun;83(2):110-2. Considerations for conducting research in agricultural biotechnology. Shelton AM. Department of Entomology, Cornell University/NYSAES, 416 Barton Laboratory, Geneva, NY 14456, USA. ams5@cornell.edu Science has shown its increased vulnerability because of two recent high-profile articles published in major journals on corn produced through biotechnology: a laboratory report suggesting profound consequences to monarch butterfly populations due to Bt corn pollen and a report suggesting transgenic introgression into Mexican maize. While both studies have been widely regarded as having flawed methodology, publishing these studies has created great consternation in the scientific community, regulatory agencies and the general public. There are roles and responsibilities of scientists, scientific journals, the public media, public agencies, and those who oppose or advocate a specific technology, and serious consequences when those roles and responsibilities go awry. Modern communication may exacerbate the flow of misinformation and easily lead to a decline in public confidence about biotechnology and science. However, common sense tells us that scientific inquiry and the publication and reporting of results should be performed with high standards of ethical behavior, regardless of one's personal perspective on agricultural biotechnology. Publication Types: Review PMID: 12788279 [PubMed - indexed for MEDLINE] 1542: J Invertebr Pathol. 2003 Jun;83(2):107-9. Regulatory and associated political issues with respect to Bt transgenic maize in the European union. Saeglitz C, Bartsch D. Chair of Biology V, Ecology, Ecochemistry and Ecotoxicology, Aachen University of Technology-RWTH Aachen, D-52056 Aachen, Germany. Legislation at the national level in Europe as well as that developed by the European Union (EU) generally permits release and commercialization of genetically modified organisms (GMOs). However, only 10 plant/event combinations were registered as of 2002: three maize events (Bt176, Mon810, and Bt11), with the other seven divided among carnation (3), oil-seed rape (2), tobacco (1), and raddiccio (1). Of these, only one maize event (Bt176) has been registered as a legal variety, and this was in Spain, where 22,000ha have been planted annually since 1998. In this paper, we first provide an overview on the complexity of EU GMO legislation. Then we discuss the minor role that results of EU-funded biosafety research have had on governmental policy. Finally, we provide information about initiatives for post-commercialization monitoring plans of Bt maize in Europe. As a result of the slow progress to date, we conclude that commercialization of GMOs will be seriously delayed in the EU for the next several years. Publication Types: Review PMID: 12788278 [PubMed - indexed for MEDLINE] 1543: J Invertebr Pathol. 2003 Jun;83(2):100-3. Legal and regulatory concerns about transgenic plants in Brazil. Fontes EM. Embrapa Recursos Genéticos e Biotecnologia, C.P. 01372/02372, Brasília, DF CEP 70849-970, Brazil. eliana@cenargen.embrapa.br Brazil has a biosafety law that was approved in 1995. This law provides for a horizontal type of regulation that coordinates other existing regulatory frameworks in the areas of agriculture, health and environment. Various federal government departments are responsible for implementing the law. The National Technical Biosafety Commission is the national competent authority on biosafety with overall responsibility. In the case of Bt plants or any insecticidal organism, the Agrochemical Law also applies and authorization for laboratory, greenhouse and field studies must be obtained from the Plant Protection Secretariat, the Brazilian Institute of Environment and the National Agency of Health. Furthermore, the National Environmental Council must issue a license for commercialization of any GMO. There is pressure needed for capacity building and to harmonize the regulatory and administrative frameworks among the different federal departments involved. Some perspectives and challenges for the commercial registration of transgenic crops are discussed. Publication Types: Review PMID: 12788276 [PubMed - indexed for MEDLINE] 1544: J Invertebr Pathol. 2003 Jun;83(2):93-9. Regulation of Bt crops in Canada. Macdonald P, Yarrow S. Canadian Food Inspection Agency, 59 Camelot Drive, Ontario, Ottawa, Canada, K1A 0Y9. pmacdonald@inspection.gc.ca The Canadian Food Inspection Agency (CFIA) regulates environmental releases of plants with novel traits, which include transgenic plants such as Bt crops. Bt crops are regulated in Canada because they express insect resistance novel to their species. Commercialization of crops with novel traits such as the production of insecticidal Bt proteins requires an approval for environmental release, as well as approvals for use as feed and food. Environmental factors such as potential impacts on non-target species are considered. Insect resistance management (IRM) may be imposed as a condition for environmental release of Bt crops to delay the development of resistance in the target insect. Bt potato and European corn borer-resistant Bt corn have been released with mandatory IRM. The CFIA imposes an IRM plan consisting of appropriate refugia, education of farmers and seed dealers, and monitoring and mitigation. Industry, regulators, government extension staff and public researchers provide expert advice on IRM. Publication Types: Review PMID: 12788275 [PubMed - indexed for MEDLINE] 1545: Plant J. 2003 Jun;34(5):647-59. A C-terminal sequence of soybean beta-conglycinin alpha' subunit acts as a vacuolar sorting determinant in seed cells. Nishizawa K, Maruyama N, Satoh R, Fuchikami Y, Higasa T, Utsumi S. Laboratory of Food Quality Design and Development, Graduate School of Agriculture, Kyoto University, Uji, Kyoto 611-0011, Japan. In maturing seed cells, many newly synthesized proteins are transported to the protein storage vacuoles (PSVs) via vesicles unique to seed cells. Vacuolar sorting determinants (VSDs) in most of these proteins have been determined using leaf, root or suspension-cultured cells apart from seed cells. In this study, we examined the VSD of the alpha' subunit of beta-conglycinin (7S globulin), one of the major seed storage proteins of soybean, using Arabidopsis and soybean seeds. The wild-type alpha' was transported to the matrix of the PSVs in seed cells of transgenic Arabidopsis, and it formed crystalloid-like structures. Some of the wild-type alpha' was also transported to the translucent compartments (TLCs) in the PSV presumed to be the globoid compartments. However, a derivative lacking the C-terminal 10 amino acids was not transported to the PSV matrix, and was secreted out of the cells, although a portion was also transported to the TLCs. The C-terminal region of alpha' was sufficient to transport a green fluorescent protein (GFP) to the PSV matrix. These indicate that alpha' contains two VSDs: one is present in the C-terminal 10 amino acids and is for the PSV matrix; and the other is for the TLC (the globoid compartment). We further verified that the C-terminal 10 amino acids were sufficient to transport GFP to the PSV matrix in soybean seed cells by using a transient expression system. Publication Types: Research Support, Non-U.S. Gov't PMID: 12787246 [PubMed - indexed for MEDLINE] 1546: Int J Parasitol. 2003 May;33(5-6):479-93. Plant-based vaccines. Streatfield SJ, Howard JA. ProdiGene, Inc., 101 Gateway Boulevard, College Station, TX 77845, USA. sstreatfield@prodigene.com Plant systems are reviewed with regard to their ability to express and produce subunit vaccines. Examples of different types of expression systems producing a variety of vaccine candidates are illustrated. Many of these subunit vaccines have been purified and shown to elicit an immune response when injected into animal models. This review also includes vaccines that have been administered orally in a non-purified form as a food or feed product. Cases are highlighted which demonstrate that orally delivered plant-based vaccines can elicit immune responses and in some case studies, confer protection. Examples are used to illustrate some of the inherent advantages of a plant-based system, such as cost, ease of scale-up and convenience of delivery. Also, some of the key steps are identified that will be necessary to bring these new vaccines to the market. Publication Types: Review PMID: 12782049 [PubMed - indexed for MEDLINE] 1547: Lancet. 2003 May 24;361(9371):1798. USA fights Europe's ban on genetically modified food. Bosch X. Publication Types: News PMID: 12781547 [PubMed - indexed for MEDLINE] 1548: Nat Biotechnol. 2003 Jun;21(6):599-601. Making 'safety first' a reality for biotechnology products. Kapuscinski AR, Goodman RM, Hann SD, Jacobs LR, Pullins EE, Johnson CS, Kinsey JD, Krall RL, La Viña AG, Mellon MG, Ruttan VW. Department of Fisheries, Wildlife and Conservation Biology, 200 Hodson Hall, 1980 Folwell Ave., St. Paul, Minnesota 55108, USA. Publication Types: Research Support, Non-U.S. Gov't PMID: 12776139 [PubMed - indexed for MEDLINE] 1549: Nat Biotechnol. 2003 Jun;21(6):598. Comment in: Nat Biotechnol. 2003 Sep;21(9):974-5; author reply 975. Comment on: Nat Biotechnol. 2003 Mar;21(3):224-6. Identity tags. Pauli U. Publication Types: Comment Letter PMID: 12776138 [PubMed - indexed for MEDLINE] 1550: J Nutr. 2003 Jun;133(6):1909-12. Safety assessment of recombinant green fluorescent protein orally administered to weaned rats. Richards HA, Han CT, Hopkins RG, Failla ML, Ward WW, Stewart CN Jr. Food Safety Center of Excellence, University of Tennessee, Knoxville, TN 37996, USA. Several proposed biotechnological applications of green fluorescent protein (GFP) are likely to result in its introduction into the food supply of domestic animals and humans. We fed pure GFP and diets containing transgenic canola expressing GFP to young male rats for 26 d to evaluate the potential toxicity and allergenicity of GFP. Animals (n = 8 per group) were fed either AIN-93G (control), control diet plus 1.0 mg of purified GFP daily, modified control diet with 200 g/kg canola (Brassica rapa cv Westar), or control diet with 200 g/kg transgenic canola containing one of two levels of GFP. Ingestion of GFP did not affect growth, food intake, relative weight of intestine or other organs, or activities of hepatic enzymes in serum. Comparison of the amino acid sequence of GFP to known food allergens revealed that the greatest number of consecutive amino acid matches between GFP and any food allergen was four, suggesting the absence of common allergen epitopes. Moreover, GFP was rapidly degraded during simulated gastric digestion. These data indicate that GFP is a low allergenicity risk and provide preliminary indications that GFP is not likely to represent a health risk. Publication Types: Research Support, U.S. Gov't, Non-P.H.S. PMID: 12771338 [PubMed - indexed for MEDLINE] 1551: Nucleic Acids Res. 2003 Jun 1;31(11):e62. A novel multiplex quantitative DNA array based PCR (MQDA-PCR) for quantification of transgenic maize in food and feed. Rudi K, Rud I, Holck A. MATFORSK, Norwegian Food Research Institute, Osloveien 1, N-1430 AAS, Norway. knut.rudi@matforsk.no We have developed a novel multiplex quantitative DNA array based PCR method (MQDA-PCR). The MQDA-PCR is general and may be used in all areas of biological science where simultaneous quantification of multiple gene targets is desired. We used quantification of transgenic maize in food and feed as a model system to show the applicability of the method. The method is based on a two-step PCR. In the first few cycles bipartite primers containing a universal 5' 'HEAD' region and a 3' region specific to each genetically modified (GM) construct are employed. The unused primers are then degraded with a single-strand DNA-specific exonuclease. The second step of the PCR is run containing only primers consisting of the universal HEAD region. The removal of the primers is essential to create a competitive, and thus quantitative PCR. Oligo nucleotides hybridising to internal segments of the PCR products are then sequence specifically labelled in a cyclic linear signal amplification reaction. This is done both to increase the sensitivity and the specificity of the assay. Hybridisation of the labelled oligonucleotides to their complementary sequences in a DNA array enables multiplex detection. Quantitative information was obtained in the range 0.1-2% for the different GM constructs tested. Seventeen different food and feed samples were screened using a twelve-plex system for simultaneous detection of seven different GM maize events (Bt176, Bt11, Mon810, T25, GA21, CBH351 and DBT418). Ten samples were GM positive containing mainly mixtures of Mon810, Bt11 and Bt176 DNA. One sample contained appreciable amounts of GA21. An eight-plex MQDA-PCR system for detection of Mon810, Bt11 and Bt176 was evaluated by comparison with simplex 5' nuclease PCRs. There were no significant differences in the quantifications using the two approaches. The samples could, by both methods, be quantified as containing >2%, between 1 and 2%, between 0.1 and 1%, or <0.1% in 43 out of 47 determinations. The described method is modular, and thus suited for future needs in GM detection. Publication Types: Evaluation Studies Research Support, Non-U.S. Gov't PMID: 12771226 [PubMed - indexed for MEDLINE] 1552: Toxicology. 2003 Jun 30;188(2-3):297-307. Safety assessment for genetically modified sweet pepper and tomato. Chen ZL, Gu H, Li Y, Su Y, Wu P, Jiang Z, Ming X, Tian J, Pan N, Qu LJ. The National Laboratory of Protein Engineering and Plant Genetic Engineering, Peking University, Beijing, 100871, China. zhchen@pku.edu.cn The coat protein (CP) gene of cucumber mosaic virus (CMV) was cloned from a Chinese CMV isolate, the CaMV promoter and NOS terminator added and the gene construct was transformed into both sweet pepper and tomato plants to confer resistance to CMV. Safety assessments of these genetically modified (GM) plants were conducted. It was found that these two GM products showed no genotoxicity either in vitro or in vivo by the micronucleus test, sperm aberration test and Ames test. Animal feeding studies showed no significant differences in growth, body weight gain, food consumption, hematology, blood biochemical indices, organ weights and histopathology between rats or mice of either sex fed with either GM sweet pepper or tomato diets compared with those with non-GM diets. These results demonstrate that the CMV-resistant sweet pepper and tomato are comparable to the non-GM counterparts in terms of food safety. Publication Types: Research Support, Non-U.S. Gov't PMID: 12767699 [PubMed - indexed for MEDLINE] 1553: Nature. 2003 May 22;423(6938):379-80. Brazilian science: Under new management. Adam D. Publication Types: News PMID: 12761518 [PubMed - indexed for MEDLINE] 1554: Nature. 2003 May 22;423(6938):369. Trade war looms as US launches challenge over transgenic crops. Knight J. Publication Types: News PMID: 12761507 [PubMed - indexed for MEDLINE] 1555: Nature. 2003 May 22;423(6938):367. Trade war: what is it good for? [No authors listed] Publication Types: Editorial PMID: 12761506 [PubMed - indexed for MEDLINE] 1556: J Natl Med Assoc. 2003 Feb;95(2):152-66. Environmental factors associated with asthma. Walker B Jr, Stokes LD, Warren R. Howard University Medical Center, Environmental Health Administration, District of Columbia Department of Health, Washington, USA. Asthma, a disease of attacks and remission, continues to account for substantial morbidity and direct economic costs. Numerous studies--epidemiologic, toxicologic and clinical--present evidence for a broad spectrum of environmental risk factors associated with asthma. This review summarizes current thinking on a subset of these factors. Knowledge of potential environmental determinants of asthma is important to both the patient and healthcare professional in the application of multiple modalities of medical and environmental intervention for management of the development, and exacerbation of this chronic inflammatory disorder of the airways. Publication Types: Review PMID: 12760611 [PubMed - indexed for MEDLINE] 1557: Appl Microbiol Biotechnol. 2003 Aug;62(2-3):140-50. Epub 2003 May 21. Bioflavoring and beer refermentation. Vanderhaegen B, Neven H, Coghe S, Verstrepen KJ, Derdelinckx G, Verachtert H. Center for Malting and Brewing Science, Katholieke Universiteit Leuven, Kasteelpark Arenberg 22, 3001, Heverlee, Belgium. bart.vanderhaegen@agr.kuleuven.ac.be Various techniques are used to adjust the flavors of foods and beverages to new market demands. Although synthetic flavoring chemicals are still widely used, flavors produced by biological methods (bioflavors) are now more and more requested by consumers, increasingly concerned with health and environmental problems caused by synthetic chemicals. Bioflavors can be extracted from plants or produced with plant cell cultures, microorganisms or isolated enzymes. This Mini-Review paper gives an overview of different systems for the microbial production of natural flavors, either de novo, or starting with selected flavor precursor molecules. Emphasis is put on the bioflavoring of beer and the possibilities offered by beer refermentation processes. The use of flavor precursors in combination with non-conventional or genetically modified yeasts for the production of new products is discussed. Publication Types: Review PMID: 12759790 [PubMed - indexed for MEDLINE] 1558: Trends Plant Sci. 2003 May;8(5):208-12. Risk assessment of GM plants: avoiding gridlock? Wilkinson MJ, Sweet J, Poppy GM. School of Plant Sciences, The University of Reading, Whiteknights, Reading, UK RG6 6AS. m.j.wilkinson@reading.ac.uk Cultivation of genetically modified crops is presently based largely on four crops containing few transgenes and grown in four countries. This will soon change and pose new challenges for risk assessment. A more structured approach that is as generic as possible is advocated to study consequences of gene flow. Hazards should be precisely defined and prioritized, with emphasis on quantifying elements of exposure. This requires coordinated effort between large, multidisciplinary research teams. PMID: 12758037 [PubMed - indexed for MEDLINE] 1559: Cancer Epidemiol Biomarkers Prev. 2003 May;12(5):419-25. Alcohol consumption, alcohol dehydrogenase 3 polymorphism, and colorectal adenomas. Tiemersma EW, Wark PA, Ocké MC, Bunschoten A, Otten MH, Kok FJ, Kampman E. Division of Human Nutrition and Epidemiology, Wageningen University, 6700 EV Wageningen, The Netherlands. Alcohol is a probable risk factor with regard to colorectal neoplasm and is metabolized to the carcinogen acetaldehyde by the genetically polymorphic alcohol dehydrogenase 3 (ADH3) enzyme. We evaluated whether the association between alcohol and colorectal adenomas is modified by ADH3 polymorphism. We recruited 433 cases with adenomatous polyps and 436 polyp-free controls among Caucasians undergoing endoscopy between 1995 and 2000. Frequency and amount of habitual alcohol consumption were assessed by beverage type, using a validated self-administered food frequency questionnaire. All participants provided blood for genotyping of ADH3. Multivariate analyses adjusting for gender, age, and indication for endoscopy showed that alcohol increased the risk of colorectal adenomas among women [odds ratio (OR), 1.8; 95% confidence interval (CI), 1.0-3.2, >/=10 versus <1 drink/week]. Among men, the risk of adenomas was increased only for those consuming > 21 drinks/week (OR, 1.8; 95% CI, 0.9-3.8, compared with men drinking < 1 drink/week). Among subjects in the highest tertile of alcohol consumption, those with the ADH3*1/*1 genotype were at higher risk (OR, 1.8; 95% CI, 1.0-3.1) than those with other ADH3 genotypes (OR, 1.2; 95% CI, 0.7-1.9) when compared with those in the lowest tertile with ADH3*1/*2 or ADH3*2/*2 genotypes. In conclusion, our findings are consistent with results of other studies, suggesting that alcohol consumption elevates the risk of adenomatous colorectal polyps. ADH3 polymorphism may modify the association between alcohol consumption and colorectal adenomas. Publication Types: Research Support, Non-U.S. Gov't PMID: 12750236 [PubMed - indexed for MEDLINE] 1560: Biomed Environ Sci. 2003 Mar;16(1):17-28. Genetically engineered corn rootworm resistance: potential for reduction of human health effects from pesticides. Oehme FW, Pickrell JA. Comparative Toxicology Laboratories, M213 Mosier Hall, 1800 Denison Ave, Kansas State University, Manhattan, KS 66506-5705, USA. oehme@vet.ksu.edu OBJECTIVE AND METHODS: Insecticide use, grower preferences regarding genetically engineered (GE) corn resistant to corn rootworm (CRW), and the health effects of using various CRW insecticides (organophosphates, pyrethroids, fipronil and carbamates) are reviewed for current and future farm practices. RESULTS: Pest damage to corn has been reduced only one-third by insecticide applications. Health costs from insecticide use appear significant, but costs attributable to CRW control are not quantifiable from available data. Methods reducing health-related costs of insecticide-based CRW control should be evaluated. As a first step, organophosphate insecticide use has been reduced as they have high acute toxicity and risk of long-term neurological consequences. A second step is to use agents which more specifically target the CRW. CONCLUSION: Whereas current insecticides may be poisonous to many species of insects, birds, mammals and humans, a protein derived from Bacillus thurigiensis and produced in plants via genetic modification can target the specific insect of CRW (Coleoptra), sparing other insect and non-insect species from injury. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 12747004 [PubMed - indexed for MEDLINE] 1561: Plant Physiol. 2003 May;132(1):36-43. Genetic modification removes an immunodominant allergen from soybean. Herman EM, Helm RM, Jung R, Kinney AJ. Plant Genetics Research Unit, United States Department of Agriculture/Agricultural Research Service, Donald Danforth Plant Science Center, 975 North Warson Street, St. Louis, Missouri 63132, USA. The increasing use of soybean (Glycine max) products in processed foods poses a potential threat to soybean-sensitive food-allergic individuals. In vitro assays on soybean seed proteins with sera from soybean-sensitive individuals have immunoglobulin E reactivity to abundant storage proteins and a few less-abundant seed proteins. One of these low abundance proteins, Gly m Bd 30 K, also referred to as P34, is in fact a major (i.e. immunodominant) soybean allergen. Although a member of the papain protease superfamily, Gly m Bd 30 K has a glycine in the conserved catalytic cysteine position found in all other cysteine proteases. Transgene-induced gene silencing was used to prevent the accumulation of Gly m Bd 30 K protein in soybean seeds. The Gly m Bd 30 K-silenced plants and their seeds lacked any compositional, developmental, structural, or ultrastructural phenotypic differences when compared with control plants. Proteomic analysis of extracts from transgenic seed detected the suppression of Gly m Bd 30 K-related peptides but no other significant changes in polypeptide pattern. The lack of a collateral alteration of any other seed protein in the Gly m Bd 30 K-silenced seeds supports the presumption that the protein does not have a role in seed protein processing and maturation. These data provide evidence for substantial equivalence of composition of transgenic and non-transgenic seed eliminating one of the dominant allergens of soybean seeds. Publication Types: Comparative Study Research Support, U.S. Gov't, Non-P.H.S. PMID: 12746509 [PubMed - indexed for MEDLINE] 1562: J Toxicol Environ Health B Crit Rev. 2003 May-Jun;6(3):211-25. Potential adverse health effects of genetically modified crops. Bakshi A. Biology Department, George Mason University, Fairfax, Virginia, USA. abakshi@gmu.edu Genetically modified crops have the potential to eliminate hunger and starvation in millions of people, especially in developing countries because the genetic modification can produce large amounts of foods that are more nutritious. Large quantities are produced because genetically modified crops are more resistant to pests and drought. They also contain greater amounts of nutrients, such as proteins and vitamins. However, there are concerns about the safety of genetically modified crops. The concerns are that they may contain allergenic substances due to introduction of new genes into crops. Another concern is that genetic engineering often involves the use of antibiotic-resistance genes as "selectable markers" and this could lead to production of antibiotic-resistant bacterial strains that are resistant to available antibiotics. This would create a serious public health problem. The genetically modified crops might contain other toxic substances (such as enhanced amounts of heavy metals) and the crops might not be "substantially equivalent" in genome, proteome, and metabolome compared with unmodified crops. Another concern is that genetically modified crops may be less nutritious; for example, they might contain lower amounts of phytoestrogens, which protect against heart disease and cancer. The review of available literature indicates that the genetically modified crops available in the market that are intended for human consumption are generally safe; their consumption is not associated with serious health problems. However, because of potential for exposure of a large segment of human population to genetically modified foods, more research is needed to ensure that the genetically modified foods are safe for human consumption. Publication Types: Review PMID: 12746139 [PubMed - indexed for MEDLINE] 1563: Proc Natl Acad Sci U S A. 2003 May 27;100(11):6855-9. Epub 2003 May 9. Control of seed germination in transgenic plants based on the segregation of a two-component genetic system. Schernthaner JP, Fabijanski SF, Arnison PG, Racicot M, Robert LS. Eastern Cereal and Oilseed Research Center, Agriculture and Agri-Food Canada, 960 Carling Avenue, Ottawa, ON, Canada K1A0C6. schernthane@agr.gc.ca We have developed a repressible seed-lethal (SL) system aimed at reducing the probability of transgene introgression into a population of sexually compatible plants. To evaluate the potential of this method, tobacco plants were transformed with an SL construct comprising gene 1 and gene 2 from Agrobacterium tumefaciens whereby gene 1 was controlled by the seed-specific phaseolin promoter modified to contain a binding site for the Escherichia coli TET repressor (R). The expression of this construct allows normal plant and seed development but inhibits seed germination. Plants containing the SL construct were crossed with plants containing the tet R gene to derive plant lines where the expression of the SL construct is repressed. Plant lines that contained both constructs allowed normal seed formation and germination, whereas seeds in which the SL construct was separated from the R gene through segregation did not germinate. The requirements of such a method to efficiently control the flow of novel traits among sexually compatible plants are discussed. Publication Types: Research Support, Non-U.S. Gov't PMID: 12740441 [PubMed - indexed for MEDLINE] 1564: J Biolaw Bus. 2000;Suppl.:67-70. Biotechnology and health. Borst E. Minister of Health, Welfare and Sports, Government of the Netherlands. PMID: 12739545 [PubMed - indexed for MEDLINE] 1565: J Biolaw Bus. 2000;Suppl.:111-5. The ethics of biotechnology. Lenoir N. French Constitutional Court, European Group on Ethics in Science and New Technologies of the European Union. PMID: 12739544 [PubMed - indexed for MEDLINE] 1566: Anal Bioanal Chem. 2003 Apr;375(8):985-93. Epub 2003 Feb 15. PCR technology for screening and quantification of genetically modified organisms (GMOs). Holst-Jensen A, Rønning SB, Løvseth A, Berdal KG. National Veterinary Institute, P.O. Box 8156 Dep., 0033, Oslo, Norway. arne.holst-jensen@vetinst.no Although PCR technology has obvious limitations, the potentially high degree of sensitivity and specificity explains why it has been the first choice of most analytical laboratories interested in detection of genetically modified (GM) organisms (GMOs) and derived materials. Because the products that laboratories receive for analysis are often processed and refined, the quality and quantity of target analyte (e.g. protein or DNA) frequently challenges the sensitivity of any detection method. Among the currently available methods, PCR methods are generally accepted as the most sensitive and reliable methods for detection of GM-derived material in routine applications.The choice of target sequence motif is the single most important factor controlling the specificity of the PCR method. The target sequence is normally a part of the modified gene construct, for example a promoter, a terminator, a gene, or a junction between two of these elements. However, the elements may originate from wildtype organisms, they may be present in more than one GMO, and their copy number may also vary from one GMO to another. They may even be combined in a similar way in more than one GMO. Thus, the choice of method should fit the purpose. Recent developments include event-specific methods, particularly useful for identification and quantification of GM content. Thresholds for labelling are now in place in many countries including those in the European Union. The success of the labelling schemes is dependent upon the efficiency with which GM-derived material can be detected. We will present an overview of currently available PCR methods for screening and quantification of GM-derived DNA, and discuss their applicability and limitations. In addition, we will discuss some of the major challenges related to determination of the limits of detection (LOD) and quantification (LOQ), and to validation of methods. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 12733008 [PubMed - indexed for MEDLINE] 1567: Curr Opin Biotechnol. 2003 Apr;14(2):238-43. Exploitation of molecular profiling techniques for GM food safety assessment. Kuiper HA, Kok EJ, Engel KH. RIKILT, Institute of Food Safety, PO Box 230, 6700 AE, Wageningen, The Netherlands. h.a.kuiper@rikit.wag-ur.nl Several strategies have been developed to identify unintended alterations in the composition of genetically modified (GM) food crops that may occur as a result of the genetic modification process. These include comparative chemical analysis of single compounds in GM food crops and their conventional non-GM counterparts, and profiling methods such as DNA/RNA microarray technologies, proteomics and metabolite profiling. The potential of profiling methods is obvious, but further exploration of specificity, sensitivity and validation is needed. Moreover, the successful application of profiling techniques to the safety evaluation of GM foods will require linked databases to be built that contain information on variations in profiles associated with differences in developmental stages and environmental conditions. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 12732328 [PubMed - indexed for MEDLINE] 1568: Curr Opin Biotechnol. 2003 Apr;14(2):221-5. Nutritional enhancement of plants. Tucker G. School of Biosciences, Sutton Bonington Campus, University of Nottingham, Loughborough, Leicester, LE12 5RD, UK. gregory.tucker@nottingham.ac.uk Plants can provide most of the nutrients required in the human diet; however, the major staple crops are often deficient in some of these nutrients. Thus, malnutrition, with respect to micronutrients like vitamin A, iron and zinc, affects >40% of the world's population. Advances in molecular biology are being exploited to produce crops enhanced in these key nutrients. Other nutritional targets include the modification of fatty acid composition and the enhancement of antioxidant levels, particularly carotenoids, such as lycopene, and flavonoids. However, the benefit of these 'biofortified' crops to human nutrition remains to be elucidated. Publication Types: Review PMID: 12732325 [PubMed - indexed for MEDLINE] 1569: J Nutr. 2003 May;133(5 Suppl 1):1490S-3S. Genetically modified plants for improved trace element nutrition. Lönnerdal B. Department of Nutrition, University of California, Davis, CA 95616, USA. bllonnerdal@ucdavis.edu Deficiencies of iron and zinc are common worldwide. Various strategies have been used to combat these deficiencies including supplementation, food fortification and modification of food preparation and processing methods. A new possible strategy is to use biotechnology to improve trace element nutrition. Genetic engineering can be used in several ways; the most obvious is to increase the trace element content of staple foods such as cereals and legumes. This may be achieved by introduction of genes that code for trace element-binding proteins, overexpression of storage proteins already present and/or increased expression of proteins that are responsible for trace element uptake into plants. However, even very high levels of expression may not substantially increase the iron and zinc contents unless many atoms of trace elements are bound per protein molecule. Another possibility is to introduce a protein that specifically enhances trace element absorption even in the presence of naturally occurring inhibitors, thus improving bioavailability. Genetically modifying plants so that their contents of inhibitors of trace element absorption such as phytate are substantially reduced is another approach. Increasing the expression of compounds that enhance trace element absorption such as ascorbic acid is also a possibility, although this has received limited attention so far. Iron absorption may be increased by higher ascorbic or citric acid content but require overexpression of enzymes that are involved in the synthetic pathways. Finally, a combination of all of these approaches perhaps complemented with conventional breeding techniques may prove successful. Publication Types: Review PMID: 12730450 [PubMed - indexed for MEDLINE] 1570: FEBS Lett. 2003 May 8;542(1-3):47-52. Isolation and characterization of a flowering plant male gametic cell-specific promoter. Singh M, Bhalla PL, Xu H, Singh MB. Plant Molecular Biology and Biotechnology Laboratory, Institute of Land and Food Resources, The University of Melbourne, 3010, Parkville, Vic., Australia. Flowering plant male gametic cell-specific gene expression has been reported recently but the regulatory elements controlling specificity of such genes expressed in generative cell and sperm cells have not been identified and studied. Here, we report the 0.8 kb promoter sequence upstream of the start of the transcription site of the generative cell-specific gene, LGC1, sufficient to regulate the expression of reporter genes in a cell-specific manner. In addition, the diphtheria toxin A-chain- (DT-A)-coding region under the control of the LGC1 promoter sequence confirmed unequivocally the lack of LGC1 expression in vegetative tissues. Transgenic tobacco plants carrying the LGC1-DT/A construct showed normal phenotype except for anthers of these plants that contained sterile and aborted pollen. Truncation and internal deletion analysis of the LGC1 promoter identified -242 bp as the minimal sequence necessary for male gametic cell-specific expression. In addition, a regulatory sequence required for determining generative cell-specific expression of LGC1 was identified. Deletion of this regulatory sequence led to loss of the generative cell specificity resulting in activation of this promoter in other tissues where it is normally repressed. Therefore, male gametic cell specificity of the LGC1 gene seems to be regulated by factors that suppress its activation in other plant cells. This is the first report of a male gametic cell-specific promoter, hence can be used as a novel tool in molecular analyses and experimental manipulation of flowering plant spermatogenesis and fertilization. Publication Types: Research Support, Non-U.S. Gov't PMID: 12729896 [PubMed - indexed for MEDLINE] 1571: Wei Sheng Yan Jiu. 2000 May 30;29(3):141-2. [Toxicity of anti-herbicide gene (BAR) transgenic rice] [Article in Chinese] Wang Y, Lai W, Chen J, Mei S. Zhejiang Academy of Medical Sciences, Hangzhou 310013, China. In order to evaluate the safety of anti-herbicide gene(BAR) transgenic rice, acute toxicity experiments, mutation experiments and a 30-day feeding test were conducted. The results were as follows: The oral LD50 for mice and rats was larger than 21.5 g/kg BW and no mutation was found. The rats consuming 16.32 and 64 g/kg BW were in good growth and development at the 30-day feeding test. The body weight, food utilization, routine blood tests, the ratio of organ weight to body weight, and patho-histological observations had no obvious change. The none effect dosage for the transgene rice was 64 g/kg. Publication Types: English Abstract PMID: 12725054 [PubMed - indexed for MEDLINE] 1572: Theor Appl Genet. 2003 Aug;107(3):528-39. Epub 2003 Apr 30. Hybridization between transgenic Brassica napus L. and its wild relatives: Brassica rapa L., Raphanus raphanistrum L., Sinapis arvensis L., and Erucastrum gallicum (Willd.) O.E. Schulz. Warwick SI, Simard MJ, Légère A, Beckie HJ, Braun L, Zhu B, Mason P, Séguin-Swartz G, Stewart CN Jr. Agriculture and Agri-Food Canada, Eastern Cereal and Oilseed Research Centre, K.W. Neatby Building, Central Experimental Farm, Ottawa, Ontario, K1A OC6, Canada. warwicks@agr.gc.ca The frequency of gene flow from Brassica napus L. (canola) to four wild relatives, Brassica rapa L., Raphanus raphanistrum L., Sinapis arvensis L. and Erucastrum gallicum (Willd.) O.E. Schulz, was assessed in greenhouse and/or field experiments, and actual rates measured in commercial fields in Canada. Various marker systems were used to detect hybrid individuals: herbicide resistance traits (HR), green fluorescent protein marker (GFP), species-specific amplified fragment length polymorphisms (AFLPs) and ploidy level. Hybridization between B. rapa and B. napus occurred in two field experiments (frequency approximately 7%) and in wild populations in commercial fields (approximately 13.6%). The higher frequency in commercial fields was most likely due to greater distance between B. rapa plants. All F(1) hybrids were morphologically similar to B. rapa, had B. napus- and B. rapa-specific AFLP markers and were triploid (AAC, 2n=29 chromosomes). They had reduced pollen viability (about 55%) and segregated for both self-incompatible and self-compatible individuals (the latter being a B. napus trait). In contrast, gene flow between R. raphanistrum and B. napus was very rare. A single R. raphanistrum x B. napus F1 hybrid was detected in 32,821 seedlings from the HR B. napus field experiment. The hybrid was morphologically similar to R. raphanistrum except for the presence of valves, a B. napus trait, in the distorted seed pods. It had a genomic structure consistent with the fusion of an unreduced gamete of R. raphanistrum and a reduced gamete of B. napus (RrRrAC, 2n=37), both B. napus- and R. raphanistrum-specific AFLP markers, and had <1% pollen viability. No hybrids were detected in the greenhouse experiments (1,534 seedlings), the GFP field experiment (4,059 seedlings) or in commercial fields in Québec and Alberta (22,114 seedlings). No S. arvensis or E. gallicum x B. napus hybrids were detected (42,828 and 21,841 seedlings, respectively) from commercial fields in Saskatchewan. These findings suggest that the probability of gene flow from transgenic B. napus to R. raphanistrum, S. arvensis or E. gallicum is very low (<2-5 x 10(-5)). However, transgenes can disperse in the environment via wild B. rapa in eastern Canada and possibly via commercial B. rapa volunteers in western Canada. Publication Types: Comparative Study Research Support, Non-U.S. Gov't PMID: 12721639 [PubMed - indexed for MEDLINE] 1573: Nat Biotechnol. 2003 May;21(5):480-1. Will we reap what biopharming sows? Miller HI. Hoover Institution, Stanford, CA, USA. miller@hoover.stanford.edu PMID: 12721561 [PubMed - indexed for MEDLINE] 1574: Nat Biotechnol. 2003 May;21(5):468-9. Europe sees sharp decline in GMO research. Mitchell P. Publication Types: News PMID: 12721556 [PubMed - indexed for MEDLINE] 1575: Cloning Stem Cells. 2003;5(1):63-9. Gene expression in cloned bovine fetal liver. Schrader AD, Iqbal MJ, Jones KL. Department of Animal Science, Food and Nutrition, Southern Illinois University, Carbondale, Illinois, USA. Nuclear transfer (NT) is a method of animal reproduction that bypasses fertilization and propagates known combinations of genes. Currently NT is an inefficient process. Attempts have been made to increase the efficiency of this procedure, but most have been deemed unsuccessful. Some problems associated with NT are unusually large birth weights, and physical abnormalities in developing liver, heart, and brain. Despite numerous studies performed on NT animals, the factors behind the anomalies remain unknown. It is possible that nuclear reprogramming is the basis of poor development rates, meaning, when the donor cells are fused with enucleated eggs the nuclei may not regain the full ability to direct cell differentiation in subsequent mitotic divisions. If reprogramming is not carried out precisely, then some genes may not be correctly expressed in NT animals. The purpose of this study was to determine if differential gene expression between the livers of NT fetuses when compared to an embryo transfer (ET) derived fetus could be detected and the genes identified. An Angus fetus at 45 d of gestation was collected and a non-clonal cell line established for use as NT donor cells. Two NT fetuses were propagated and compared to the original. Differential Display Reverse Transcription Polymerase Chain Reaction (ddRT-PCR) was used to identify genes that were differentially expressed. Differentially abundant cDNAs were subcloned, sequenced and their corresponding mRNAs were verified by semi-quantitative RT-PCR. Twenty-three Expressed Sequence Tags (ESTs) were sequenced in Bos taurus and submitted to GenBank. The results of ddRT-PCR identified 39 genes/ESTs that were potentially differentially expressed. Fifteen of the genes were tested by semi-quantitative RT-PCR, but no significant differences were detected. Publication Types: Research Support, Non-U.S. Gov't PMID: 12713702 [PubMed - indexed for MEDLINE] 1576: J Exp Bot. 2003 May;54(386):1317-9. Genetically modified soybeans and food allergies. Herman EM. Plant Genetics Unit, USDA/ARS, Donald Danforth Plant Science Center, 975 N. Warson Road, St Louis, MO 63132, USA. eherman@danforthcenter.org Allergenic reactions to proteins expressed in GM crops has been one of the prominent concerns among biotechnology critics and a concern of regulatory agencies. Soybeans like many plants have intrinsic allergens that present problems for sensitive people. Current GM crops, including soybean, have not been shown to add any additional allergenic risk beyond the intrinsic risks already present. Biotechnology can be used to characterize and eliminate allergens naturally present in crops. Biotechnology has been used to remove a major allergen in soybean demonstrating that genetic modification can be used to reduce allergenicity of food and feed. This provides a model for further use of GM approaches to eliminate allergens. PMID: 12709477 [PubMed - indexed for MEDLINE] 1577: Vaccine. 2003 May 16;21(17-18):2065-72. Neutralising immunogenicity of a polyepitope antigen expressed in a transgenic food plant: a novel antigen to protect against measles. Bouche FB, Marquet-Blouin E, Yanagi Y, Steinmetz A, Muller CP. Laboratoire National de Santé, Department of Immunology, WHO Collaborative Center for Measles, 20A rue Auguste Lumière, L-1950 Luxembourg, Luxembourg. Transgenic carrot plants were developed expressing a designer polyepitope combining tandem repeats of a protective loop-forming B cell epitope (H386-400) of the measles virus hemagglutinin protein with a human promiscuous, measles-unrelated T cell epitope (tt830-844). Despite the sensitivity of the loop conformation to its molecular environment, proper folding was confirmed by conformation-dependent monoclonal antibodies. The antibodies also reacted with the boiled antigen in Western blot. Immunisation of mice peritoneally with carrot plant extracts induced high titers of antibodies that crossreacted strongly with the virus. Furthermore, the sera neutralised field isolates of different geographic origins and genotypes in a modified plaque reduction neutralisation assay performed on CD150-transfected Vero cells. These results demonstrate that transgenic carrot plants can serve as an efficient expression system to produce highly immunogenic, randomly assembled polyepitope antigens. The combined features of the selected epitopes and the potential of the plant expression system may pave the way towards new vaccines against measles. Publication Types: Research Support, Non-U.S. Gov't PMID: 12706696 [PubMed - indexed for MEDLINE] 1578: Parassitologia. 2002 Dec;44(3-4):131-5. An alternative focus in strategic research on disease vectors: the potential of genetically modified non-biting mosquitoes. Coluzzi M, Costantini C. Dipartimento di Scienze di Sanità Pubblica, Sezione di Parassitologia, Università La Sapienza, Piazzale Aldo Moro 5, 00185 Roma, Italy. mario.coluzzi@uniroma1.it We examine the constraints and the feasibility of field experiments involving the release of genetically modified (GM) pathogen-resistant mosquitoes, and whether there are alternatives to the research line based on the production of refractory strains. The production of a GM mosquito strain characterized instead by obligate primiparous and parous autogeny and by disrupted host seeking and biting behaviour could make the release more acceptable by the general public. Genetic transformation should act in this case to reverse some of the essential steps of the evolutionary process that gave rise to hematophagy. The replacement strategy could be based on the mass release of both sexes in a well defined ecological niche made temporarily empty of the natural population, thus avoiding the problems related to the need of sexual competitiveness of the released material. This option is encouraged by the growing evidence that competitive exclusion mechanisms influence the pattern of distribution of different taxa within Anopheles gambiae s.s. and by the fact that the plesiomorphic characteristics of vitellogenesis without a blood meal (autogeny), which exploits fat body reserve accumulated during larval life and food other than blood in adult life, persist as genetic variants in various hematophagous insect groups, and it has been found secondarily fixed in others showing stable reversions to primiparous and parous autogeny. If this has been the result of natural selection, then the artificial production of non-biting mosquito strains, by selection and/or transgenesis, should be feasible. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 12701373 [PubMed - indexed for MEDLINE] 1579: J Ind Microbiol Biotechnol. 2003 May;30(5):302-7. Epub 2003 Apr 17. Evaluation of a high temperature immobilised enzyme reactor for production of non-reducing oligosaccharides. Schiraldi C, Di Lernia I, Giuliano M, Generoso M, D'Agostino A, De Rosa M. Department of Experimental Medicine, Section of Biotechnology and Molecular Biology, Second University of Naples, via De Crecchio n degrees 7, 80138, Naples, Italy. chiara.schiraldi@unina2.it There is interest in the production of non-reducing carbohydrates due to their potential application in various industrial fields, particularly the food industry. In this paper, we describe the development of an immobilised cell bioprocess for the synthesis of non-reducing maltodextrins at high temperatures. The trehalosyl-dextrins-forming enzyme (TDFE) isolated from the thermoacidophilic archaeon Sulfolobus solfataricus (strain MT4), was recently expressed at high yields in Escherichia coli (strain Rb-791). Here, we evaluate different matrices, such as polyacrylamide gel, crude egg white, chitosan and calcium alginate for their effectiveness in immobilising whole recombinant E. coli cells subjected to prior thermal permeabilisation. Calcium-alginate based gels formed a solid biocatalyst with a good activity yield and the best enzymatic stability at the operating temperature (75 degrees C). Therefore, these beads were used to pack a glass column reactor to perform the bioconversion of interest. Optimal operating parameters were defined in relation to the substrate stream flow-rate and the substrate-to-biocatalyst ratio. The production of trehalosylmaltotetraose from maltohexaose reached equilibrium with a constant of about 2.6 at 75 degrees C. The bioreactor was exploited for production of trehalosylmaltodextrins from a commercial mixture of maltodextrins, achieving a productivity of 106.5 mg ml(-1) h(-1) (g biocatalyst)(-1) with ~40% conversion when using a 30% (w/v) solution. Publication Types: Evaluation Studies PMID: 12700952 [PubMed - indexed for MEDLINE] 1580: Toxicol Sci. 2003 May;73(1):8-16. Epub 2003 Apr 15. Workshop overview: approaches to the assessment of the allergenic potential of food from genetically modified crops. Ladics GS, Holsapple MP, Astwood JD, Kimber I, Knippels LM, Helm RM, Dong W. The DuPont Co., Haskell Laboratory, Newark, Delaware 19714, USA. gregory.s.ladics@usa.dupont.com There is a need to assess the safety of foods deriving from genetically modified (GM) crops, including the allergenic potential of novel gene products. Presently, there is no single in vitro or in vivo model that has been validated for the identification or characterization of potential food allergens. Instead, the evaluation focuses on risk factors such as source of the gene (i.e., allergenic vs. nonallergenic sources), physicochemical and genetic comparisons to known allergens, and exposure assessments. The purpose of this workshop was to gather together researchers working on various strategies for assessing protein allergenicity: (1) to describe the current state of knowledge and progress that has been made in the development and evaluation of appropriate testing strategies and (2) to identify critical issues that must now be addressed. This overview begins with a consideration of the current issues involved in assessing the allergenicity of GM foods. The second section presents information on in vitro models of digestibility, bioinformatics, and risk assessment in the context of clinical prevention and management of food allergy. Data on rodent models are presented in the next two sections. Finally, nonrodent models for assessing protein allergenicity are discussed. Collectively, these studies indicate that significant progress has been made in developing testing strategies. However, further efforts are needed to evaluate and validate the sensitivity, specificity, and reproducibility of many of these assays for determining the allergenicity potential of GM foods. Publication Types: Congresses PMID: 12700419 [PubMed - indexed for MEDLINE] 1581: J Agric Food Chem. 2003 Apr 23;51(9):2447-56. Erratum in: J Agric Food Chem. 2004 May 19;52(10):3210. Metabolite profiling of tomato (Lycopersicon esculentum) using 1H NMR spectroscopy as a tool to detect potential unintended effects following a genetic modification. Le Gall G, Colquhoun IJ, Davis AL, Collins GJ, Verhoeyen ME. Institute of Food Research, Norwich Research Park, Colney, Norwich NR4 7UA, United Kingdom. The maize transcription factors LC and C1 were simultaneously overexpressed in tomato with the aim of producing lines with increased amounts of flavonols. The metabolite composition of these genetically modified tomatoes has been compared with that of azygous (nonmodified) controls grown side-by-side under the same conditions. It has been possible to observe metabolic changes in both types at different stages of maturity. (1)H NMR spectra showed that the levels of glutamic acid, fructose, and some nucleosides and nucleotides gradually increase from the immature to the ripe stage, whereas some amino acids such as valine and gamma-aminobutyric acid were present in higher amounts in unripe tomatoes. Apart from the significantly increased content of six main flavonoid glycosides (mainly kaempferol-3-O-rutinoside, with additional increases in kaempferol-3,7-di-O-glucoside (1), kaempferol-3-O-rutinoside-7-O-glucoside (2), kaempferol-3-O-glucoside, a dihydrokaempferol-O-hexoside (3), and naringenin-7-O-glucoside), the levels of at least 15 other metabolites were found to be different between the two types of red tomato. Among them were citric acid, sucrose, phenylalanine, and trigonelline. However, although statistically significant, these changes in mean values were relatively minor (less than 3-fold) and within the natural variation that would be observed in a field-grown crop. Nevertheless, this study clearly showed that NMR combined with chemometrics and univariate statistics can successfully trace even small differences in metabolite levels between plants and therefore represents a powerful tool to detect potential unintended effects in genetically modified crops. Publication Types: Research Support, Non-U.S. Gov't PMID: 12696919 [PubMed - indexed for MEDLINE] 1582: J Agric Food Chem. 2003 Apr 23;51(9):2438-46. Characterization and content of flavonoid glycosides in genetically modified tomato (Lycopersicon esculentum) fruits. Le Gall G, DuPont MS, Mellon FA, Davis AL, Collins GJ, Verhoeyen ME, Colquhoun IJ. Institute of Food Research, Norwich Research Park, Colney, Norwich NR4 7UA, United Kingdom. There is a growing interest in producing food plants with increased amounts of flavonoids because of their potential health benefits. Tomatoes contain small amounts of flavonoids, most of which are located in the peel of the fruit. It has been shown that flavonoid accumulation in tomato flesh, and hence an overall increase in flavonoid levels in tomato fruit, can be achieved by means of simultaneous overexpression of the maize transcription factors LC and C1. Fruit from progeny of two modified lines (2027 and 2059) was selected for a detailed analysis and individual identification of flavonoids, at different stages of maturity. Nine major flavonoids were detected in the flesh of transgenic ripe tomatoes. LC/NMR, LC/MS, and LC/MS/MS enabled us to identify these as kaempferol-3,7-di-O-glucoside (1), kaempferol-3-O-rutinoside-7-O-glucoside (2), two dihydrokaempferol-O-hexosides (3 and 4), rutin (5), kaempferol-3-O-rutinoside (6), kaempferol-3-O-glucoside (7), naringenin-7-O-glucoside (8) and naringenin chalcone (9), which were quantified by HPLC/DAD. All but 5, 6, and 9 were detected in tomato for the first time. The total flavonoid glycoside content of ripe transgenic tomatoes of line 2059 was about 10-fold higher than that of the controls, and kaempferol glycosides accounted for 60% of this. Kaempferol glycosides comprised around 5% of the flavonoid glycoside content of ripe control tomatoes (the rest was rutin and naringenin chalcone). The rutin concentration in both transgenic and control fruits was similar. Publication Types: Research Support, Non-U.S. Gov't PMID: 12696918 [PubMed - indexed for MEDLINE] 1583: J Am Vet Med Assoc. 2003 Apr 1;222(7):926. Bioengineered pigs go to market. Kuehn BM. Publication Types: News PMID: 12685776 [PubMed - indexed for MEDLINE] 1584: Biomol Eng. 2003 Mar;20(3):107-12. Food-grade expression of human glutathione S-transferase and Cu/Zn superoxide dismutase in Lactococcus lactis. Xiang H, Wei W, Tan H. Institute of Microbiology, Chinese Academy of Sciences, Beijing 100080, China. A food-grade gene expression system in Lactococcus lactis was established by the combination of a vector containing the lacF gene as the selection marker and a strain WZ103 carrying an in-frame deletion of this gene in the chromosome as the host. The human glutathione S-transferase A1-1 (hGSTA1) and Cu/Zn superoxide dismutase (hSOD) genes were respectively cloned into a food-grade vector under the control of the lactococcal inducible promoter P(lacA). The resulting expression plasmids were separately introduced into the lactose-deficient (Lac(-)) host, and the lactose-utilizing (Lac(+)) transformants were directly selected on a chemically defined medium, using lactose as the sole carbon source. The successful food-grade expression of hGSTA1 and hSOD in the L. lactis WZ103 transformed with these plasmids were analyzed by Western blotting and enzymatic activity assay, respectively. Publication Types: Evaluation Studies Research Support, Non-U.S. Gov't PMID: 12684072 [PubMed - indexed for MEDLINE] 1585: Toxicol Lett. 2003 Apr 11;140-141:297-302. Assessment of the allergenic potential of proteins. Kimber I, Betts CJ, Dearman RJ. Syngenta Central Toxicology Laboratory, Alderley Park, Cheshire SK10 4TJ, Macclesfield, UK. ian.kimber@syngenta.com The development of novel foods, including foods derived from genetically modified plants, has generated considerable interest in the design and application of appropriate safety assurance measures. A specific focus of attention has been on allergenicity, and in particular the need to determine whether the products of novel genes introduced into food plants have the potential to cause allergic sensitisation. Among the approaches applied currently are considerations of whether a new protein has structural, sequence and/or antigenic similarities with known food allergens, and whether or not it displays resistance to digestion within a simulated gastric fluid, or by pepsin. Although such data are useful in an overall hazard assessment, they are neither individually, nor collectively, able to provide a direct evaluation of inherent sensitising potential. For this reason there is a need to develop and apply appropriate animal models that will offer a more holistic view of sensitising activity. Several methods have been suggested, but as yet none has been evaluated fully or validated. Nevertheless, significant progress has been made and in this article an experimental approach using BALB/c strain mice in which animals are exposed to the test protein via systemic (intraperitoneal, or in certain circumstances, intradermal) administration is described. Inherent sensitising potential is measured as a function of induced IgE antibody responses. Experience to date is encouraging and the data available reveal that this method is able to distinguish between proteins of different allergenic potential. Publication Types: Review PMID: 12676477 [PubMed - indexed for MEDLINE] 1586: Tex Med. 2003 Mar;99(3):66-9. Safety of foods derived from genetically modified plants. Thomas JA. University of Texas Health Science Center at San Antonio, TX, USA. jat-tox@swbell.net Biopharmaceuticals have been available for clinical use for nearly three decades, but foods derived from agribiotechnology have been available for just under a decade. Controversy surrounding foods from genetically modified (GM) plants has focused primarily upon their allergenicity, with lesser concerns about antibiotic resistance genes. Concerns are related to possible environmental impacts on non-human species, including effects on non-target species (e.g., butterflies) and on the development of so-called "super weeds." Food allergies are no more prevalent in foods from GM plants than in conventional foods. Further, the use of antibiotics in the development of GM plants does not pose a significant risk to the human population. Foods from the current GM plant products have been shown not to pose any detrimental effects to humans, and, in fact, nutritionally enhanced products are being developed. GM foods are subjected globally to intense regulatory scrutiny, and extensive data have been provided consistently to regulatory agencies in the United States on a voluntary basis, with mandatory reporting of data soon to be in force. Existing environmental concerns appear to be unjustified on the basis of existing data and experience. PMID: 12674981 [PubMed - indexed for MEDLINE] 1587: J Physiol Pharmacol. 2003 Mar;54(1):81-8. Enhanced food and water intake in renin transgenic rats. Szczepańska-Sadowska E, Paczwa P, Dobruch J. Department of Experimental and Clinical Physiology, Medical University of Warsaw, Warsaw, Poland. eszs@amwaw.edu.pl In short term experiments angiotensin II (Ang II) is a potent stimulant of thirst, however it is not known whether prolonged activation of the renin-angiotensin system is associated with chronic alteration of water or food intake. Renin transgenic rats TGRmRen(2)27 (TGR) exhibit significant elevation of AngII in the brain regions involved in regulation of body fluid balance. The purpose of the present study was to find out whether TGR rats manifest also different water (WI) and food (FI) intake and renal excretory functions in comparison to their parent Sprague Dawley (SD) strain. To this end 24 h WI and FI as well as urine excretion (Vu) and urinary outputs of solutes (Cosm), sodium (UNaV) and potassium (UKV) were compared under baseline conditions in 16 TGR and 15 SD rats having free access to water and food. In 15 TGR and 17 SD rats effect of 24 h dehydration on water intake was investigated. Under baseline conditions TGR rats consumed significantly greater amount of food and water than SD rats. Vu, UNaV and UKV were not significantly different in both strains. Cumulative water intakes in SD and TGR rats subjected to 24 h dehydration did not differ. The results reveal that under baseline conditions TGR rats manifest greater food and water intakes than SD rats whereas stimulation of thirst by water deprivation is similar in both strains. The results suggest that the ingestive behavior may be chronically altered by upregulation of the renin-angiotensin system. Publication Types: Research Support, Non-U.S. Gov't PMID: 12674220 [PubMed - indexed for MEDLINE] 1588: Bull Acad Natl Med. 2002;186(8):1391-400. [Allergic risk of transgenic food: prevention strategies] [Article in French] Moneret-Vautrin DA. Service de Médecine Interne-Immunologie Clinique et Allergologie, Hôpital Central, 29 avenue du Maréchal de Lattre de Tassigny, 54035 Nancy. Numerous allergens proceed from foods. The allergic risk of transgenic foods needs to be evaluated according recommendations from the Joint Expert Committee FAO/WHO. Potential issues are the risk of cross reactivity with existing allergens, the modification of allergenicity of the transgenic protein induced by a modified metabolism in the host, the modified allergenicity of the proteins of the transgenic plant, a potential neo-allergenicity of the transgenic protein, and the risk of dissemination through pollens, inducing a respiratory sensitization then a cross food allergy. The algorithm includes three steps for evaluation: first the search for significant homology of the protein with allergens listed in allergen databanks, or the identity of a sequence of six aminoacids with known allergens, then a cross reactivity explored through the binding to IgEs from patients allergic to the source of the gene, or allergic to organisms of the same group or botanical family, and finally the extent of the pepsine resistance. The risk of immunogenicity has to be studied with appropriate animal models. A post-marketing surveillance is recommended for monitoring of adverse effects. The structure of an Allergo-Vigilance Network, the tools for efficiency and the groups at higher risk will be discussed. Publication Types: English Abstract Review PMID: 12669358 [PubMed - indexed for MEDLINE] 1589: Bull Acad Natl Med. 2002;186(8):1377-88; discussion 1388-9. [GMOs in food: risk assessment and management; scientific and regulatory characteristics] [Article in French] Casse F, Hervieu F. Université Montpellier 2, Biochimie et Physiologie Moléculaire des Plantes, UMR 5004 CNRS/UM2/INRA/Agro-M), Place Viala, 34060 Montpellier. Genetic transformation constitutes a new tool for improvement of microorganisms, animals and plants used in food. Foreseeable risks are evoked, as well as management measures to avoid GMO unsuspected risks. Few risks are specific to GMOs. Present elements of french and european regulations concerning placing on the market and follow up GMOs and other novel foods are described. Publication Types: English Abstract PMID: 12669357 [PubMed - indexed for MEDLINE] 1590: Ann Pharm Fr. 2003 Mar;61(2):124-31. [Summary and conclusions] [Article in French] Rérat A. Académie nationale de médecine, Paris. PMID: 12668951 [PubMed - indexed for MEDLINE] 1591: Ann Pharm Fr. 2003 Mar;61(2):96-102. [The allergic risk of transgenic foods strategy for prevention] [Article in French] Moneret-Vautrin DA. Service de Médecine Interne-Immunologie Clinique et Allergologie, Hôpital Central, 29 avenue du Maréchal de Lattre de Tassigny, F54035 Nancy Cedex. A significant number of allergens arise from foods. The allergic risk of transgenic foods must be evaluated in accordance with the recommendations of the Joint Expert Committee FAO/WHO. Potential issues are the risk of cross reactivity with existing allergens, the modification of allergenicity of the transgenic protein induced by a modified metabolism in the host, the modified allergenicity of the proteins of the transgenic plant, a potential neo-allergenicity of the transgenic protein, and the risk of dissemination through pollens, inducing a respiratory sensitization then a cross food allergy. The algorithm includes three steps for evaluation: first the search for significant homology of the protein with allergens listed in allergen databanks, or the identity of a sequence of six aminoacids with known epitopes, then a cross reactivity explored through the binding to IgEs from patients allergic to the source of the gene, or allergic to organisms of the same group or botanical family, and finally the extent of the pepsine resistance. The risk of immunogenicity has to be studied with appropriate animal models. A post-marketing surveillance is recommended for monitoring of adverse effects. The structure of an Allergo-Vigilance Network, the tools for efficiency and the groups at higher risk will be discussed. The potential risk of transgenic foods to be allergenic cannot be overlooked, not ignoring the fact that current technologies modify allergenicity of foods. Publication Types: English Abstract Review PMID: 12668947 [PubMed - indexed for MEDLINE] 1592: Ann Pharm Fr. 2003 Mar;61(2):87-95. [GMOs in food: risk assessment, scientific management and regulatory aspects] [Article in French] Casse F, Hervieu F. Université Montpellier 2, CC 024, place Eugène Bataillon, F34095 Montpellier Cedex 5. Genetic transformation constitutes a new tool for improvement of microorganisms, animals and plants used in food. We present foreseeable risks, as well as management measures to avoid unsuspected risks of GMOs. Few risks are specific to GMOs. Present elements of French and European regulations concerning placing on the market and follow up GMOs and other novel foods are described. Publication Types: English Abstract PMID: 12668946 [PubMed - indexed for MEDLINE] 1593: Ann Pharm Fr. 2003 Mar;61(2):78-85; discussion 85-6. [Transgenic plants: towards a more balanced diet?] [Article in French] Chupeau Y. INRA, Versailles-Grignon, Biologie Cellulaire, route de Saint-Cyr, F 78026 Versailles Cedex. Over the past ten years, knowledge of the metabolic capacities of plants has expanded considerably. Combined with technical achievements in gene transfer, a better understanding of metabolic regulation provides new more precise and targeted means of modifying plant products in order to improve health and well being through diet. While a great number of attempts to modify plant metabolism are still in the exploratory phase, some key applications are emerging with applications involving micro and macronutrients. Publication Types: English Abstract Review PMID: 12668945 [PubMed - indexed for MEDLINE] 1594: Ann Pharm Fr. 2003 Mar;61(2):75-7. [Joint session of the National Academy of Medicine and the National Academy of Pharmacy, November 26, 2002] [Article in French] Rérat A; National Academy of Medicine; National Academy of Pharmacy. Académie nationale de médecine, Paris. PMID: 12668944 [PubMed - indexed for MEDLINE] 1595: Nat Biotechnol. 2003 Apr;21(4):366-9. Who is driving biotechnology acceptance? Kalaitzandonakes N, Bijman J. Agrobiotechnology Center, University of Missouri-Columbia, USA. KalaitzandonakesN@missouri.edu PMID: 12665818 [PubMed - indexed for MEDLINE] 1596: Nat Biotechnol. 2003 Apr;21(4):346-7. US food aid still under GM cloud. Jayaraman KS. Publication Types: News PMID: 12665809 [PubMed - indexed for MEDLINE] 1597: Nat Biotechnol. 2003 Apr;21(4):346. Italy employs further GMO delay tactics. Meldolesi A. Publication Types: News PMID: 12665808 [PubMed - indexed for MEDLINE] 1598: Anal Bioanal Chem. 2003 Mar;375(6):799-804. Epub 2003 Feb 27. Erratum in: Anal Bioanal Chem. 2003 Jul;376(5):763-4. Evaluation of glycoalkaloids in tubers of genetically modified virus Y-resistant potato plants (var. Désirée) by non-aqueous capillary electrophoresis coupled with electrospray ionization mass spectrometry (NACE-ESI-MS). Bianco G, Schmitt-Kopplin P, Crescenzi A, Comes S, Kettrup A, Cataldi TR. Dipartimento di Chimica, Università degli Studi della Basilicata, Via N. Sauro 85, 85100 Potenza, Italy. The glycoalkaloid content of transgenic potatoes was evaluated by an optimised method based on non-aqueous capillary electrophoresis coupled on-line with electrospray ionization-mass spectrometry (NACE-ESI-MS). The potato material consisted of tubers from a conventional cv. Désirée and from three lines of modified plants resistant, intermediate and susceptible to infection by potato virus Y (PVY). The main glycoalkaloids were confirmed to be alpha-solanine and alpha-chaconine with parent ion masses m/z 852 and 868, respectively. In addition, an unknown minor peak at m/z 850.6 was found both in conventional (control) and susceptible line potato tubers. Such a compound exhibited an MS(2) spectrum with fragments ions at 704 and 396 m/z derived by loss of two ions, i.e. m/z 146 and 307, most likely corresponding to a rhamnose unit and a [glucose-(rhamnose)(2)] moiety, respectively. Up to 30-80-fold higher concentrations of total glycoalkaloids were found in the peel compared to flesh samples of all tubers examined. TGA content was nearly doubled in peel samples of resistant compared to control lines, and these levels were lower than the limit recommended for food safety, i.e. 20-60 mg of TGA per 100 g fresh weight. Moreover, it was established that tubers produced by virus-resistant clones are substantially equivalent in glycoalkaloid contents to those produced by conventional potato varieties. Publication Types: Research Support, Non-U.S. Gov't PMID: 12664181 [PubMed - indexed for MEDLINE] 1599: Plant J. 2003 Apr;34(1):1-11. The redistribution of protein sulfur in transgenic rice expressing a gene for a foreign, sulfur-rich protein. Hagan ND, Upadhyaya N, Tabe LM, Higgins TJ. CSIRO Plant Industry, GPO Box 1600, Canberra ACT 2601, Australia. Sulfur amino acid composition is an important determinant of seed protein quality. A chimeric gene encoding sunflower seed albumin (SSA), one of the most sulfur-rich seed storage proteins identified so far, was introduced into rice (Oryza sativa) in order to modify cysteine and methionine content of the seed. Analysis of a transgenic line expressing SSA at approximately 7% of total seed protein revealed that the mature grain showed little change in the total sulfur amino acid content compared to the parental genotype. This result indicated that the transgenic rice grain was unable to respond to the added demand for cysteine and methionine imposed by the production of SSA. Analysis of the protein composition of the transgenic grain showed changes in the relative levels of the major seed storage proteins, as well as some non-storage proteins, compared to non-transgenic controls. Changes observed at the protein level were concomitant with differences in mRNA accumulation but not always with the level of transcription. The limited sulfur reserves appeared to be re-allocated from endogenous proteins to the new sulfur sink in the transgenic grain. We hypothesize that this response is mediated by a signal transduction pathway that normally modulates seed storage protein composition in response to environmental fluctuations in sulfur availability, via both transcriptional and post-transcriptional control of gene expression. PMID: 12662304 [PubMed - indexed for MEDLINE] 1600: Mol Cells. 2003 Feb 28;15(1):20-6. The 5-enolpyruvylshikimate-3-phosphate synthase of glyphosate-tolerant soybean expressed in Escherichia coli shows no severe allergenicity. Chang HS, Kim NH, Park MJ, Lim SK, Kim SC, Kim JY, Kim JA, Oh HY, Lee CH, Huh K, Jeong TC, Nam DH. College of Pharmacy, Yeungnam University, Gyongsan 712-749, Korea. The recombinant gene was amplified from the chromosomal DNA of genetically-modified (GM) soybeans and identified as epsps encoding 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) which renders glyphosate resistance. The epsps structural gene was introduced in the pET28(a) plasmid for its expression in Escherichia coli BL21(DE3). It was confirmed that the maximal productivity of the EPSPS protein was achieved when cultivating the recombinant strain in a LB broth for 2 h after supplementing 1 mM isopropylbeta-D-thiogalactopyranoside (IPTG) in a 2 h-culture broth. Since the expressed EPSPS protein was found as an insoluble form in the inclusion body, it was extracted by 6 M urea after sonication, and then purified through immobilized nickel-affinity column chromatography to isolate EPSPS having a molecular mass of 57 kDa. When incubated in simulated gastric fluid containing pepsin at pH 1.5, the purified EPSPS protein was completely digested within 1 min. In addition, the passive cutaneous anaphylaxis reaction of the purified EPSPS protein was not observed in the Sprague Dawley rat system that was administered either orally or subcutaneously. Furthermore, treatment of the EPSPS protein to the culture of the sensitized peritoneal mast cells, or unsensitized but antisera-labeled mast cells, showed neither a remarkable change in the histamine release nor a cytokine production, including interleukin-4 (IL-4) and tumor necrosis factor-alpha (TNF-alpha). Thus, it can be concluded that the EPSPS protein in the GM soybean showed no significant allergenicity in the Sprague Dawley rats. Publication Types: Research Support, Non-U.S. Gov't PMID: 12661756 [PubMed - indexed for MEDLINE] 1601: St Thomas Law Rev. 2001 Summer;13(4):927-34. A Protestant perspective on the new genetics: are people and institutions of faith prepared to lead? Jones JC. Plymouth Congregational Church Foundation, Inc., USA. PMID: 12661582 [PubMed - indexed for MEDLINE] 1602: Biochem Soc Trans. 2003 Apr;31(2):299-306. Genes in food--why the furore? Dixon B. 130 Cornwall Road, Ruislip Manor, Middx. HA4 6AW, U.K. Although unprecedented and perhaps unique in its irrationality, the recent furore over genetically modified (GM) food holds extremely important lessons for scientists. Some sections of the media undoubtedly bear a heavy responsibility for giving the expression 'GM' threatening connotations that are quite unwarranted. However, influential contributions to the hysteria have come from a surprising range of other sources, including some scientists. The research community has failed in its responsibility to society in three ways. Firstly, plant scientists did not appreciate that certain techniques (such as the use of antibiotic resistance genes as markers during plant transformation) would inevitably provoke public consternation. As a result, they took no steps to address such concerns. Secondly, researchers overlooked, minimized or in some cases simply dismissed the significance of public fears that they were 'interfering with Nature' or 'playing God'. Thirdly, plant breeders apparently saw no need to take pro-active measures with regard to the media and public in placing potential environmental and nutritional benefits of GM crops on the agenda in a positive fashion. Partly because of this failure, GM food is now firmly established in the public mind as wholly objectionable. One measure of how far we have travelled down that road is that it hardly matters any more whether objections are based on alleged environmental risks of cultivating GM crops or alleged toxicological hazards of eating them. 'Genetically modified organism', like 'radioactivity', has become an odious, generic shibboleth. Given that millions of people throughout the world are already benefiting from pharmaceuticals made by GM organisms, this is bizarre. Publication Types: Historical Article Lectures PMID: 12653625 [PubMed - indexed for MEDLINE] 1603: Nature. 2003 Mar 20;422(6929):246. Biotech firms join charities in drive to help Africa's farms. Hoag H. Publication Types: News PMID: 12646881 [PubMed - indexed for MEDLINE] 1604: Metab Eng. 2002 Oct;4(4):263-72. Genetic engineering of a zeaxanthin-rich potato by antisense inactivation and co-suppression of carotenoid epoxidation. Römer S, Lübeck J, Kauder F, Steiger S, Adomat C, Sandmann G. Universität Konstanz, Lehrstuhl für Physiologie und Biochemie der Pflanzen, Fachbereich Biologie, Germany. Zeaxanthin is an important dietary carotenoid but its abundance in our food is low. In order to provide a better supply of zeaxanthin in a staple crop, two different potato (Solanum tuberosum L.) varieties were genetically modified. By transformation with sense and antisense constructs encoding zeaxanthin epoxidase, zeaxanthin conversion to violaxanthin was inhibited. Both approaches (antisense and co-suppression) yielded potato tubers with higher levels of zeaxanthin. Depending on the transgenic lines and tuber development, zeaxanthin content was elevated 4 to 130-fold reaching values up to 40 microg/g dry weight. As a consequence of the genetic manipulation, the amount of violaxanthin was diminished dramatically and in some cases the monoepoxy intermediate antheraxanthin accumulated. Between one and eight copies of the sense or antisense epoxidase gene fragments were integrated into the genome. In addition, most of the transformants with higher zeaxanthin levels showed also increased total carotenoid contents (up to 5.7-fold) and some of them exhibited reduced amounts of lutein. The increase in total carotenoids suggests that the genetic modification affects the regulation of the whole carotenoid biosynthetic pathway in potato tubers. Northern blot analysis demonstrated that upregulation of carotenogenesis in the transgenics is accompanied by substantial higher phytoene synthase transcript levels in 6-week-old tubers and a very slight increase of the beta-carotene hydroxylase transcript. The amount of the deoxyxylulose 5-phosphate synthase mRNA was very similar in wild type and transformed tubers. Abscisic acid content of tubers remained unchanged whereas alpha-tocopherol was 2 to 3 fold elevated in the transformants. Publication Types: Comparative Study Research Support, Non-U.S. Gov't PMID: 12646321 [PubMed - indexed for MEDLINE] 1605: Cumberland Law Rev. 2001-2002;32(3):475-86. Who owns my DNA?: the national and international intellectual property law on human embryonic tissue and cloning. Knowles SM, Adams SD. PMID: 12645552 [PubMed - indexed for MEDLINE] 1606: Proc Biol Sci. 2003 Feb 22;270(1513):335-40. A novel approach to the use of genetically modified herbicide tolerant crops for environmental benefit. Dewar AM, May MJ, Woiwod IP, Haylock LA, Champion GT, Garner BH, Sands RJ, Qi A, Pidgeon JD. Broom's Barn Research Station, Higham, Bury St Edmunds, Suffolk IP28 6NP, UK. alan.dewar@bbsrc.ac.uk The proposed introduction of genetically modified herbicide tolerant (GMHT) crops, with claims of improved weed control, has prompted fears about possible environmental impacts of their widespread adoption, particularly on arable weeds, insects and associated farmland birds. In response to this, we have developed a novel weed-management system for GMHT sugar beet, based on band spraying, which exploits the flexibility offered by the broad-spectrum partner herbicides. Here, we show the results from two series of field experiments which, taken together, demonstrate that, by using this system, crops can be managed for enhanced weed and insect biomass without compromising yield, thus potentially offering food and shelter to farmland birds and other wildlife. These results could be applicable widely to other row crops, and indicate that creative use of GMHT technology could be a powerful tool for developing more sustainable farming systems in the future. Publication Types: Evaluation Studies Research Support, Non-U.S. Gov't PMID: 12639311 [PubMed - indexed for MEDLINE] 1607: Shokuhin Eiseigaku Zasshi. 2002 Dec;43(6):339-47. [Comparison of soyasaponin and isoflavone contents between genetically modified (GM) and non-GM soybeans] [Article in Japanese] Goda Y, Akiyama H, Suyama E, Takahashi S, Kinjo J, Nohara T, Toyoda M. National Institute of Health Sciences: 1-18-1, Kamiyoga, Setagaya-ku, Tokyo 158-8501, Japan. Soyasaponins and isoflavones are main secondary metabolites in soybeans. In this report we compared the content of secondary metabolites between genetically modified (GM) and non-GM soybeans. Six cultivars/lines of GM and six cultivars/lines of non-GM soybeans were extracted with methanol. Each extract was partitioned with aqueous methanol and hexane and the aqueous methanol fraction was partially purified by HP-20 and LH-20 column chromatography to afford crude soyasaponin and isoflavone fractions. The main A-type soyasaponin, acetylsoyasaponin A1 (AcA1), and the main B-type soyasaponins, soyasaponins I and II (I and II), in the crude fractions were identified by LC/MS analyses with authentic samples. The main isoflavones, daidzin, genistin, daidzein and genistein (DI, GI, DE and GE), in the crude fractions were identified by LC photo-diode array analyses with authentic samples. The contents of AcA1, I and II in the crude soyasaponin fractions and those of DI, GI, DE and GE in the crude isoflavone fractions were analyzed by reversed-phase HPLC. The average contents (mg/100 g) of AcA1, I, II and total of B-type soyasaponins in GM soybeans were 36.4 +/- 24.2, 51.2 +/- 11.8, 26.4 +/- 7.6 and 77.7 +/- 18.5, respectively, and those in non-GM ones were 22.3 +/- 14.7, 46.3 +/- 17.8, 19.8 +/- 9.1 and 65.9 +/- 26.9, respectively. The average contents (mg/100 g) of DI, GI, DE, GE and total isoflavones in GM soybeans were 93.1 +/- 15.5, 121.8 +/- 19.4, 0.073 +/- 0.178, 0.320 +/- 0.082 and 215.3 +/- 33.3, respectively, and those in non-GM ones were 78.8 +/- 34.6, 106.7 +/- 28.3, 2.206 +/- 4.468, 0.822 +/- 0.754 and 188.5 +/- 26.7, respectively. There were no statistically significant differences in soyasaponin and isoflavone contents between GM and non-GM soybeans. Therefore, it was estimated that the GM soybeans are equivalent to the non-GM ones in terms of the composition of the main secondary metabolites. Publication Types: Comparative Study English Abstract PMID: 12635335 [PubMed - indexed for MEDLINE] 1608: EMBO Rep. 2003 Mar;4(3):229-32. Feeding prejudice. Reluctance within the European Union to accept genetically modified crops may hinder the benefits of this technology reaching the developing world. Owens SR. PMID: 12634834 [PubMed - indexed for MEDLINE] 1609: Nature. 2003 Mar 13;422(6928):111-2. Chinese agribiotech: Against the grain. Macilwain C. Publication Types: News PMID: 12634752 [PubMed - indexed for MEDLINE] 1610: Nature. 2003 Mar 13;422(6928):103. Tougher rules aim to prevent gene flow into crops. Hoag H. PMID: 12634744 [PubMed - indexed for MEDLINE] 1611: Nature. 2003 Mar 13;422(6928):99. A little protectionism goes a long way. [No authors listed] Publication Types: Editorial PMID: 12634739 [PubMed - indexed for MEDLINE] 1612: Appetite. 2003 Feb;40(1):9-14. The perceived healthiness of functional foods. A conjoint study of Danish, Finnish and American consumers' perception of functional foods. Bech-Larsen T, Grunert KG. The MAPP Centre, The Aarhus School of Business, Haslegaardsvej 10, DK-8210, Aarhus V, Denmark. tib@asb.dk Functional foods presumably enable the consumer to lead a healthier life without changing eating habits. Whether consumers accept this proposition or not is potentially influenced by their perceptions of the healthiness of the processing methods, enrichment components, food-types, and health claims used in the production and marketing of functional foods. Because consumers may perceive functional enrichment as interfering with nature, cultural values pertaining to man's manipulation of nature may also influence consumer acceptance of functional foods.The purpose of the study described here is to clarify to which extent Danish, Finnish and American consumers' perceptions of the healthiness of functional foods are explained by the factors mentioned above. The general results indicate that values pertaining to man's manipulation of nature is only modestly related to the acceptance of functional foods, whereas the use of different health claims, processing methods, enrichments, product types, and especially the interactions between the two latter, are important determinants of consumers' perceptions of the healthiness of functional foods. Publication Types: Comparative Study PMID: 12631500 [PubMed - indexed for MEDLINE] 1613: Mutat Res. 2003 Feb-Mar;523-524:183-92. Development and application of test methods for the detection of dietary constituents which protect against heterocyclic aromatic amines. Kassie F, Sundermann VM, Edenharder R, Platt KL, Darroudi F, Lhoste E, Humbolt C, Muckel E, Uhl M, Kundi M, Knasmüller S. Institute of Cancer Research, University of Vienna, Borschkegasse 8a, A-1090 Vienna, Austria. This article describes the development and use of assay models in vitro (genotoxicity assay with genetically engineered cells and human hepatoma (HepG2) cells) and in vivo (genotoxicity and short-term carcinogenicity assays with rodents) for the identification of dietary constituents which protect against the genotoxic and carcinogenic effects of heterocyclic aromatic amines (HAs). The use of genetically engineered cells expressing enzymes responsible for the bioactivation of HAs enables the detection of dietary factors that inhibit the metabolic activation of HAs. Human derived hepatoma (HepG2) cells are sensitive towards HAs and express several enzymes [glutathione S-transferase (GST), N-acetyltransferase (NAT), sulfotransferase (SULT), UDP-glucuronosyltransferase (UDPGT), and cytochrome P450 isozymes] involved in the biotransformation of HAs. Hence these cells may reflect protective effects, which are due to inhibition of activating enzymes and/or induction of detoxifying enzymes. The SCGE assay with rodent cells has the advantage that HA-induced DNA damage can be monitored in a variety of organs which are targets for tumor induction by HAs. ACF and GST-P(+) foci constitute preneoplastic lesions that may develop into tumors. Therefore, agents that prevent the formation of these lesions may be anticarcinogens. The foci yield and the sensitivity of the system could be substantially increased by using a modified diet. The predictive value of the different in vitro and in vivo assays described here for the identification of HA-protective dietary substances relevant for humans is probably better than that of conventional in vitro test methods with enzyme homogenates. Nevertheless, the new test methods are not without shortcomings and these issues are critically discussed in the present article. Copyright 2002 Elsevier Science B.V. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 12628516 [PubMed - indexed for MEDLINE] 1614: Nat Biotechnol. 2003 Apr;21(4):439-42. Epub 2003 Mar 10. A transformation method for obtaining marker-free plants of a cross-pollinating and vegetatively propagated crop. de Vetten N, Wolters AM, Raemakers K, van der Meer I, ter Stege R, Heeres E, Heeres P, Visser R. AVEBE, AVEBE-weg 1, 9607 PT Foxhol, The Netherlands. It is generally thought that transformation of plant cells using Agrobacterium tumefaciens occurs at a very low frequency. Therefore, selection marker genes are used to identify the rare plants that have taken up foreign DNA. Genes encoding antibiotic and herbicide resistance are widely used for this purpose in plant transformation. Over the past several years, consumer and environmental groups have expressed concern about the use of antibiotic- and herbicide-resistance genes from an ecological and food safety perspective. Although no scientific basis has been determined for these concerns, generating marker-free plants would certainly contribute to the public acceptance of transgenic crops. Several methods have been reported to create marker gene-free transformed plants, for example co-transformation, transposable elements, site-specific recombination, or intrachromosomal recombination. Not only are most of these systems time-consuming and inefficient, but they are also employed on the assumption that isolation of transformants without a selective marker gene is not feasible. Here we present a method that permits the identification of transgenic plants without the use of selectable markers. This strategy relies on the transformation of tissue explants or cells with a virulent A. tumefaciens strain and selection of transformed cells or shoots after PCR analysis. Incubation of potato explants with A. tumefaciens strain AGL0 resulted in transformed shoots at an efficiency of 1-5% of the harvested shoots, depending on the potato genotype used. Because this system does not require genetic segregation or site-specific DNA-deletion systems to remove marker genes, it may provide a reliable and efficient tool for generating transgenic plants for commercial use, especially in vegetatively propagated species like potato and cassava. Publication Types: Evaluation Studies Research Support, Non-U.S. Gov't PMID: 12627169 [PubMed - indexed for MEDLINE] 1615: Appl Environ Microbiol. 2003 Mar;69(3):1623-8. Improved anaerobic use of arginine by Saccharomyces cerevisiae. Martin O, Brandriss MC, Schneider G, Bakalinsky AT. Department of Food Science and Technology, Oregon State University, Corvallis, OR 97331-6602, USA. Anaerobic arginine catabolism in Saccharomyces cerevisiae was genetically modified to allow assimilation of all four rather than just three of the nitrogen atoms in arginine. This was accomplished by bypassing normal formation of proline, an unusable nitrogen source in the absence of oxygen, and causing formation of glutamate instead. A pro3 ure2 strain expressing a PGK1 promoter-driven PUT2 allele encoding Delta(1)-pyrroline-5-carboxylate dehydrogenase lacking a mitochondrial targeting sequence produced significant cytoplasmic activity, accumulated twice as much intracellular glutamate, and produced twice as much cell mass as the parent when grown anaerobically on limiting arginine as sole nitrogen source. Publication Types: Evaluation Studies Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. PMID: 12620851 [PubMed - indexed for MEDLINE] 1616: J Environ Sci Health B. 2003 Mar;38(2):211-9. Extractable soil lipids and microbial activity as affected by Bt and non Bt maize grown on a silty clay loam soil. Dinel H, Schnitzer M, Saharinen M, Meloche F, Paré T, Dumontet S, Lemee L, Ambles A. Eastern Cereal and Oilseed Research Centre, Agriculture and Agri-Food Canada, Ottawa, Canada. denise.drd@sympatico.ca Pyrolysis-gas (Py-GC) chromatography was used to characterize extractable lipids from Bt and non-Bt maize shoots and soils collected at time of harvesting. Py-GC-MS (mass spectrometry) showed that the concentrations of total alkenes identified in non-Bt shoots and soils were 47.9 and 21.3% higher than in Bt maize shoots and soils, respectively. N-alkanes identified were of similar orders of magnitude in Bt and non-Bt maize shoots, but were 28.6% higher in Bt than in non-Bt soils. Bt maize shoots contained 29.7% more n-fatty acids than non-Bt maize shoots, whereas the concentrations of n-fatty acids in Bt soils were twice as high as those in non-Bt soils. Concentrations of unsaturated fatty acids in Bt maize shoots were 22.1% higher than those in non-Bt maize shoots, while concentrations of unsaturated fatty acids were 22.5% higher in non-Bt than in Bt soils. The cumulative CO2-C evolved from soils under Bt and non-Bt crops was 30.5% lower under Bt as compared to non-Bt crops, whereas when maize shoots were added to Bt and non-Bt soils, the decrease in CO2-C evolved were 16.5 and 23.6%, respectively. Our data showed that the cultivation of Bt maize significantly increased the saturated to unsaturated lipid ratios in soils which appeared to negatively affect microbial activity. PMID: 12617558 [PubMed - indexed for MEDLINE] 1617: In Silico Biol. 2002;2(4):525-34. Prediction of food protein allergenicity: a bioinformatic learning systems approach. Zorzet A, Gustafsson M, Hammerling U. Division of Toxicology, National Food Administration, P.O. Box 622, SE-751 26 Uppsala, Sweden. Food hypersensitivity is constantly increasing in Western societies with a prevalence of about 1-2% in Europe and in the USA. Among children, the incidence is even higher. Because of the introduction of foods derived from genetically modified crops on the marketplace, the scientific community, regulatory bodies and international associations have intensified discussions on risk assessment procedures to identify potential food allergenicity of the newly introduced proteins. In this work, we present a novel biocomputational methodology for the classification of amino acid sequences with regard to food allergenicity and non-allergenicity. This method relies on a computerised learning system trained using selected excerpts of amino acid sequences. One example of such a successful learning system is presented which consists of feature extraction from sequence alignments performed with the FASTA3 algorithm (employing the BLOSUM50 substitution matrix) combined with the k-Nearest-Neighbour (kNN) classification algorithm. Briefly, the two features extracted are the alignment score and the alignment length and the kNN algorithm assigns the pair of extracted features from an unknown sequence to the prevalent class among its k nearest neighbours in the training (prototype) set available. 91 food allergens from several specialised public repositories of food allergy and the SWALL database were identified, pre-processed, and stored, yielding one of the most extensively characterised repositories of allergenic sequences known today. All allergenic sequences were classified using a standard one-leave-out cross validation procedure yielding about 81% correctly classified allergens and the classification of 367 non-allergens in an independent test set resulted in about 98% correct classifications. The biocomputational approach presented should be regarded as a significant extension and refinement of earlier attempts suggested for in silico food safety assessment. Our results show that the framework described here is powerful enough to become useful as part of a multiple-procedure test scheme that also depicts other evaluation approaches such as solid phase immunoassay and tests for stability to digestions. Publication Types: Research Support, Non-U.S. Gov't PMID: 12611632 [PubMed - indexed for MEDLINE] 1618: Nature. 2003 Feb 13;421(6924):689. Concern about Japan's unclear biotech regulations. Watanabe KN. Publication Types: Letter PMID: 12610594 [PubMed - indexed for MEDLINE] 1619: Nature. 2003 Feb 13;421(6924):675. Missing the big picture. [No authors listed] Publication Types: Editorial PMID: 12610579 [PubMed - indexed for MEDLINE] 1620: Nat Biotechnol. 2003 Mar;21(3):224-6. Comment in: Nat Biotechnol. 2003 Jun;21(6):598. Encoding technical information in GM organisms. Marillonnet S, Klimyuk V, Gleba Y. Icon Genetics AG, Maximilianstr. 38/40, Munich 80539, Germany. PMID: 12610560 [PubMed - indexed for MEDLINE] 1621: Shokuhin Eiseigaku Zasshi. 2002 Oct;43(5):301-5. [A detection method for recombinant DNA from genetically modified potato (NewLeaf Y potato)] [Article in Japanese] Akiyama H, Watanabe T, Wakui C, Chiba Y, Shibuya M, Goda Y, Toyoda M. National Institute of Health Sciences: 1-18-1, Kamiyoga, Setagaya-ku, Tokyo 158-8501, Japan. A detection method using the polymerase chain reaction (PCR) was developed to detect genetically modified (GM) potato (NewLeaf Y potato; NL-Y), of which the mandatory assessment has not yet been completed in Japan. The potato sucrose synthase gene was used as an internal control. We designed a primer pair to specifically detect NL-Y without false-positive results in processed potato foods infected with the potato virus Y (PVY). The DNA introduced into NL-Y using the primer pair could be detected from potato powder samples containing 0.05% NL-Y. In addition, we designed primer pairs for recognizing the CryIIIA gene to detect the NewLeaf potato (NL), NewLeaf Plus potato (NL-P) and NL-Y and for recognizing p-FMV in order to detect NL-P and NL-Y. The proposed method was applied to the detection of NL-Y in 26 processed potato foods and NL-Y was not detected in any samples. Publication Types: English Abstract Research Support, Non-U.S. Gov't PMID: 12607929 [PubMed - indexed for MEDLINE] 1622: Shokuhin Eiseigaku Zasshi. 2002 Oct;43(5):273-9. Effect of subchronic feeding of genetically modified corn (CBH351) on immune system in BN rats and B10A mice. Teshima R, Watanabe T, Okunuki H, Isuzugawa K, Akiyama H, Onodera H, Imai T, Toyoda M, Sawada J. National Institute of Health Sciences: 1-18-1, Kamiyoga, Setagaya-ku, Tokyo 158-8501, Japan. Subchronic animal feeding studies to examine the effect on the immune system of genetically modified corn CBH351, which contains the Cry9C protein derived from Bacillus thuringiensis subspecies tolworthi, were conducted in female BN rats and B10A mice. The studies were designed to compare the effect of a line of genetically modified corn CBH351 (GM corn) with that of isoline corn (non-GM corn). Heat-treated corn meal was incorporated into the diets of the rats and mice at a concentration of 50%. The study duration was 13 weeks. Growth, food intake, and organ weights of the thymus, spleen, and liver were compared between animals fed the non-GM and GM lines. The histological findings in thymus, spleen, mesenteric lymph nodes, Peyer's patches, small intestines, liver, kidney, and bone marrow, and the presence of Cry9C-specific IgE, IgG, IgG1 and IgA antibodies in serum were also compared. The results showed no significant differences in growth, feeding value, or the histological findings in immunity-related organs between the animals fed the GM and non-GM lines. Production of Cry9 C-specific IgE and IgA was not detected in the serum of either group. Production of Cry9C-specific IgG and IgG1 was slightly increased in the 50% GM groups of BN rats. No Cry9C-specific IgG or IgG1 was detected in the serum of BN rats fed the diet containing 5% GM-corn In conclusion, no immunotoxic activity was detected in the GM-corn-fed rats and mice in this subchronic dietary study. Publication Types: Research Support, Non-U.S. Gov't PMID: 12607925 [PubMed - indexed for MEDLINE] 1623: Nature. 2003 Feb 27;421(6926):889. Comment on: Nature. 2003 Jan 23;421(6921):304. Objective assessment of transgenic salmon. Gray G. Publication Types: Comment Letter PMID: 12606971 [PubMed - indexed for MEDLINE] 1624: Plant Mol Biol. 2003 Jan;51(2):213-24. Abnormal 'wrinkled' cell walls and retarded development of transgenic Arabidopsis thaliana plants expressing endo-1,4-beta-glucanase (cell) antisense. Tsabary G, Shani Z, Roiz L, Levy I, Riov J, Shoseyov O. The Kennedy Leigh Centre for Horticultural Research and The Otto Warburg Center for Agricultural Biotechnology, The Faculty of Agricultural, Food and Environmental Quality Sciences, The Hebrew University of Jerusalem, PO. Box 12, Rehovot 76100, Israel. Transgenic Arabidopsis thaliana plants expressing cel1 antisense exhibit reduced levels of cel1 mRNA and protein compared with wild-type plants. The former display significant alterations in their phenotype. cel1 antisense plants have shorter stems and roots and are mechanically weaker than their wild-type counterparts. In cel1 antisense plants, the cell wall structure is markedly disrupted: both fluorescent confocal microscopy and scanning electron microscopy revealed 'wrinkled' cell walls, thus indicating that CEL1 plays an important role in cell wall relaxation during cell growth and expansion. In cel1 antisense plants, the number of xylem elements per bundle is smaller than in the wild-type. In addition, both xylem elements and interfascicular fibers are significantly less lignified in the former. It is suggested that in A. thaliana, abnormal cell wall deposition affected by CEL1 depletion is associated not only with cell growth, but also with the differentiation process in the vascular and supporting tissues. PMID: 12602880 [PubMed - indexed for MEDLINE] 1625: Plant Mol Biol. 2003 Feb;51(3):351-62. The cryptic enhancer elements of the tCUP promoter. Wu K, Hu M, Martin T, Wang C, Li XQ, Tian L, Brown D, Miki B. Eastern Cereal and Oilseed Research Centre, Agriculture and Agri-Food Canada, Ottawa, Ontario Canada K1A 0C6. Examination of the tCUP cryptic promoter from tobacco demonstrates that cryptic gene regulatory elements in the plant genome are functionally equivalent to elements responsible for the expression of plant genes. They are also organized in a similar fashion. Analysis of the expression pattern of the GUS reporter gene in transgenic Arabidopsis plants revealed that all of the information needed for strong constitutive expression was located in the truncated, -394tCUP promoter fragment. A series of 5' deletion and linker-scan mutagenesis constructs identified two separate enhancer elements. A long AT-rich region was identified between positions -350 and -161 bp relative to the transcription start site. 5' deletions that removed this A/T-rich fragment resulted in a significant decrease in promoter activity; whereas, oligomerization enhanced activity. A 21 bp sequence (TAGCCCCAATTTCAAATTCAA) spanning nucleotides -150 to -130 relative to transcription start site was also identified in a similar fashion and defined a novel cryptic constitutive enhancer element (Cce). Electrophoretic mobility-shift assays showed that tobacco nuclear proteins that interacted strongly with the tCUP promoter bound specifically to the 21-bp Cce element, suggesting that this sequence is probably a binding site(s) for transcription factors. The Cce element was dependent on the AT-rich element for activity indicating combinatorial control. The combined effects of the A/T rich and Cce elements appear to be responsible for the constitutive transcriptional activity of the tCUP promoter. Publication Types: Research Support, Non-U.S. Gov't PMID: 12602866 [PubMed - indexed for MEDLINE] 1626: Ned Tijdschr Geneeskd. 2003 Jan 11;147(2):56-60. Comment on: Ned Tijdschr Geneeskd. 2003 Jan 11;147(2):60-5. [Nutrition and health--genetically modified food] [Article in Dutch] Kuiper HA, Kleter GA, Kok EJ. Wageningen Universiteit & Research Centre, Rijks-Kwaliteitsinstituut voor Land- en Tuinbouwproducten, Bornsesteeg 45, 6708 PD Wageningen. h.a.kuiper@rikilt.wag-ur.nl The genetically modified (GM) crops cultivated at present have new properties of benefit to agriculture. It is expected that in the future GM crops will also be cultivated with more complex genetic modifications that are aimed at improving the nutritional and health value to the consumer. The safety assessment of GM foods before market approval is based on a comparison of the characteristics of the GM food with those of the conventional counterpart. Identified differences are thoroughly tested for their toxicological and nutritional consequences. Supplementary modern analytical techniques are being developed for the assessment of future complex GM foods. No cases of adverse health or nutritional effects in consumers have been reported for the existing generation of GM foods. The feasibility of post-market surveillance of (GM) foods, in order to identify small or chronic effects that have not been noticed in the pre-market phase, is being investigated, yet its value should not be overestimated. Surveillance can be informative in case of specific questions concerning certain products as long as the consumer intake is well documented. To this end traceability and labelling systems must be set up. Publication Types: Comment English Abstract Research Support, Non-U.S. Gov't Review PMID: 12602068 [PubMed - indexed for MEDLINE] 1627: J Food Prot. 2003 Feb;66(2):304-10. Model studies on the detectability of genetically modified feeds in milk. Poms RE, Hochsteiner W, Luger K, Glössl J, Foissy H. Department of Dairy Science and Bacteriology, University of Agricultural Sciences Vienna, A-1180 Vienna, Austria roland.poms@jrc.it Detecting the use of genetically modified feeds in milk has become important, because the voluntary labeling of milk and dairy products as "GMO free" or as "organically grown" prohibits the employment of genetically modified organisms (GMOs). The aim of this work was to investigate whether a DNA transfer from foodstuffs like soya and maize was analytically detectable in cow's milk after digestion and transportation via the bloodstream of dairy cows and, thus, whether milk could report for the employment of transgene feeds. Blood, milk, urine, and feces of dairy cows were examined, and foreign DNA was detected by polymerase chain reaction by specifically amplifying a 226-bp fragment of the maize invertase gene and a 118-bp fragment of the soya lectin gene. An intravenous application of purified plant DNA showed a fast elimination of marker DNA in blood or its reduction below the detection limit. With feeding experiments, it could be demonstrated that a specific DNA transfer from feeds into milk was not detectable. Therefore, foreign DNA in milk cannot serve as an indicator for the employment of transgene feeds unless milk is directly contaminated with feed components or airborne feed particles. Publication Types: Research Support, Non-U.S. Gov't PMID: 12597493 [PubMed - indexed for MEDLINE] 1628: Biochimie. 2002 Nov;84(11):1073-87. Genetically modified lactic acid bacteria: applications to food or health and risk assessment. Renault P. Génétique microbienne, Inra, domaine de Vilvert, 78352 Jouy-en-Josas, France. renault@jouy.inra.fr Lactic acid bacteria have a long history of use in fermented food products. Progress in gene technology allows their modification by introducing new genes or by modifying their metabolic functions. These modifications may lead to improvements in food technology (bacteria better fitted to technological processes, leading to improved organoleptic properties em leader ), or to new applications including bacteria producing therapeutic molecules that could be delivered by mouth. Examples in these two fields will be discussed, at the same time evaluating their potential benefit to society and the possible risks associated with their use. Risk assessment and expected benefits will determine the future use of modified bacteria in the domains of food technology and health. Publication Types: Review PMID: 12595135 [PubMed - indexed for MEDLINE] 1629: Nature. 2003 Feb 20;421(6925):775. Ministers back gene-crop advisers. Butler D. PMID: 12594469 [PubMed - indexed for MEDLINE] 1630: Biochem Biophys Res Commun. 2003 Feb 28;302(1):133-7. Reduction of antigenicity and allergenicity of genetically modified egg white allergen, ovomucoid third domain. Mine Y, Sasaki E, Zhang JW. Department of Food Science, University of Guelph, Ont., Guelph, Canada N1G2W1. ymine@uoguelph.ca Ovomucoid (Gal d1) is a major allergen in hen egg white, consisting of three tandem domains. In this study, five genetically modified third domain (DIII) mutants, which were substituted single or double amino acids within its IgE and IgG epitopes were compared with those prepared and their antigenicity and allergenicity with native analogue using Western immunoblot and enzyme-linked immunosorbent assay. The replacement of phenylalanine at 37 (F37) position with methionine caused drastical loss of IgG and IgE binding activities of human sera derived from egg allergic patients as well as disruption of the alpha-helix structure which comprises a part of the IgG and IgE epitopes. Substituting glycine at 32 position in conjunction with F37 showed a synergistic effect of decreasing antigenicity. The present study indicated that glycine 32 and phenylalanine 37 have an important role on its antigenicity and allergenicity as well as structural integrity of ovomucoid DIII. Publication Types: Research Support, Non-U.S. Gov't PMID: 12593859 [PubMed - indexed for MEDLINE] 1631: Croat Med J. 2003 Feb;44(1):102-6. Risks and benefits of genetically modified maize donations to southern Africa: views from Malawi. Muula AS, Mfutso-Bengo JM. University of Malawi College of Medicine, Department of Public Health, Private Bag 360, Chichiri, Blantyre 3, Malawi. amuula@commhealth.medcol.mw In 2001 and 2002, many countries in the Southern African Development Community (SADC) have suffered from severe food shortages resulting in an estimated 14 million people facing starvation due to inadequate quantities of the staple maize. The international community's response has been the donation of foodstuffs, including genetically modified maize. Reactions of the recipient countries of Zambia, Zimbabwe, and Malawi have been different. Zambia appealed to the donors not to send genetically modified maize, whereas Malawi accepted the maize donations. Malawi is currently facing many public health challenges because 10% of its 10-million population is HIV-positive, maternal mortality rate has almost doubled between 1992 and 2000, and there are also an estimated 1 million orphans due to HIV/AIDS. In the European Union, genetically modified maize falls under "Novel Foods" and its marketing and distribution are strictly regulated by law. This has never been the case in the southern African countries. In this article, we discuss the ethical challenges associated with genetically modified maize donations to southern Africa. Although genetically modified food offers a way to avoid many adverse effects of food shortages, we believe that some of the ethical questions of genetically modified food donations should be solved first, under the leadership of the donor countries and partnership of the developing countries. There are fears that consummation of genetically modified maize could have adverse health effects. These fears must be addressed if the confidence of developing countries in the donor community is to be maintained. PMID: 12590438 [PubMed - indexed for MEDLINE] 1632: Rapid Commun Mass Spectrom. 2003;17(5):479-83. Analysis of protein profiles of genetically modified potato tubers by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Careri M, Elviri L, Mangia A, Zagnoni I, Agrimonti C, Visioli G, Marmiroli N. Dipartimento di Chimica Generale ed Inorganica, Chimica Analitica, Chimica Fisica, Università degli Studi di Parma, Italy. Traceability of genetically modified (GM) foods demands the development of appropriate reliable techniques in order to identify and quantify peptide or nucleic acid residues in GM plants and food products through the food chain. In this study the applicability of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) was demonstrated for the characterization of proteins of transformed and untransformed potato (Solanum Tuberosum L.) tubers. In GM tubers the expression level of the G1-1 gene, which regulates transition from dormancy to sprouting tubers, was inhibited by antisense technology. The analysis of antisense transformed lines showed that several of them exhibited a significant delay in sprouting relative to the control lines, in accordance with a decrease in the transcript level. Preliminary attempts to compare the protein patterns obtained from transformed and control lines using traditional electrophoresis were not able to reveal differences in the low-kDa range. Instead, MALDI-TOFMS applied to total peptide extract without any purification was able to distinguish spectral patterns of transformed and untransformed lines. In particular, several characteristic peaks from m/z 4373 to 4932 were detected only in the mass spectra of GM tuber samples. Copyright 2003 John Wiley & Sons, Ltd. Publication Types: Research Support, Non-U.S. Gov't PMID: 12590397 [PubMed - indexed for MEDLINE] 1633: Phytochemistry. 2003 Mar;62(6):939-47. Identification and quantification of carotenoids, tocopherols and chlorophylls in commonly consumed fruits and vegetables. Burns J, Fraser PD, Bramley PM. School of Biological Sciences, Royal Holloway, University of London, Egham Surrey TW20 0EX, UK. The carotenoid, tocopherol and chlorophyll metabolic profiles and content of a selection of fruits and vegetables found commonly in the diet, have been determined using a rapid RP-HPLC technique with on-line PDA detection. Information gathered from the screening of secondary plant metabolites is vital for the accurate determination of the dietary intake of these micro-nutrients, and in the development of comprehensive food tables. Determination of basal levels is also necessary for the rational engineering of health-promoting phytochemicals in food crops. In addition this approach can also be applied to the routine screening of products to determine metabolic differences between varieties and cultivars, as well as between genetically modified and the corresponding non-genetically modified tissue. PMID: 12590121 [PubMed - indexed for MEDLINE] 1634: J Am Diet Assoc. 2003 Feb;103(2):208-14. Consumer perception of risk associated with eating genetically engineered soybeans is less in the presence of a perceived consumer benefit. Brown JL, Ping Y. Department of Food Science, Pennsylvania State University, University Park, PA 16802, USA. f9a@psu.edu OBJECTIVE: To determine whether perceived benefit alters personal risk perception associated with eating genetically engineered soybeans, consumer desire for labeling, preferred phrase on a label symbol, and desired information in an educational brochure. DESIGN: Comparison of responses of two consumer groups who completed one of two survey versions. SUBJECTS/SETTINGS: One hundred fifty supermarket shoppers, age 21 years and older, for each survey or n=300 total. Focus groups and a pilot test were used to develop the final survey in which consumers read a description of a genetically engineered soybean with either no obvious consumer benefit or an obvious consumer benefit and then completed a set of attitude questions and evaluated a voluntary label design and educational brochure content. Main outcome measures were mean opinion scores of personal risk and desire for labeling and ranking of desired label phrase and brochure topics. STATISTICAL ANALYSIS: Chi;(2) and t Tests were used. RESULTS: Consumers reading about the soybean with obvious consumer benefit were significantly more comfortable eating these than those reading about the soybean with no obvious consumer benefit (2.9+/-1.1 vs 3.4+/-1.0, respectively; Por=35% identity in an overlap of >or=80 amino acids, but only 6 of the 7 were similar across the length of the protein, or shared >50% identity to an allergen. CONCLUSIONS: An evaluation of a protein by the FASTA algorithm is the most predictive of a clinically relevant cross-reactive allergen. An additional search for matches of 8 amino acids may provide an added margin of safety when assessing the potential allergenicity of a protein, but a search with a 6-amino-acid window produces many random, irrelevant matches. Copyright 2002 S. Karger AG, Basel Publication Types: Comparative Study Evaluation Studies PMID: 12218366 [PubMed - indexed for MEDLINE] 1741: Int Arch Allergy Immunol. 2002 Aug;128(4):271-9. Allergies to cross-reactive plant proteins. Latex-fruit syndrome is comparable with pollen-food allergy syndrome. Yagami T. Division of Medical Devices, National Institute of Health Sciences, Kamiyoga, Setagaya-Ku, Tokyo, Japan. yagami@nihs.go.jp Both latex-fruit syndrome and oral allergy syndrome concomitant with pollinosis (pollen-food allergy syndrome) are considered to be caused by cross-reactivity between sensitizers and symptom elicitors. The cross-reactive food allergens relevant to these syndromes are mostly sensitive to heat and digestive enzymes. Such a vulnerable antigen cannot sensitize people perorally but provokes allergic reactions in already sensitized patients based on its cross-reactivity to the corresponding sensitizer. These types of food allergens are often called incomplete food allergens or nonsensitizing elicitors. Their features contrast with those of complete food allergens that have the capacity for peroral sensitization as well as symptom elicitation. Although highly antigenic and cross-reactive, carbohydrate epitopes do not generally elicit allergic reactions and often disturb in vitro IgE tests. Recent research has revealed that some of the cross-reactive allergens responsible for the two syndromes are proteins related to the defense responses of higher plants. Plant defense-related proteins are relatively conserved in the course of evolution and can supply cross-reactive epitopes. It is important to note that various stresses can stimulate the expression of these proteins, which implies that allergens increase in plants under stressful conditions like severe growing situations and exposure to some kinds of chemicals. Because defense-related proteins usually provide a plant with resistance to stresses, varieties that are apt to intensively induce such proteins are agriculturally valuable. Less toxic substances that cause crops to express defensive proteins are being investigated as a new type of agrochemical. Moreover, some defense-related proteins are going to be constantly produced in genetically modified plants. Even though these proteins can be useful agriculturally, their allergenicity should be evaluated carefully. Copyright 2002 S. Karger AG, Basel Publication Types: Comparative Study Review PMID: 12218365 [PubMed - indexed for MEDLINE] 1742: Nature. 2002 Sep 5;419(6902):7. Gene-bank expansion plan launched at Earth summit. Cherry M. Publication Types: News PMID: 12214203 [PubMed - indexed for MEDLINE] 1743: Nat Biotechnol. 2002 Sep;20(9):871. Comment on: Nat Biotechnol. 2002 Jun;20(6):527. Beyond gene containment. Johnson B, Dallimore R. Publication Types: Comment Letter PMID: 12205499 [PubMed - indexed for MEDLINE] 1744: Nat Biotechnol. 2002 Sep;20(9):868. Labeling GM foods--the ethical way forward. Reiss M. Science Education, University of London Institute of Education, UK. m.reiss@ioe.ac.uk PMID: 12205497 [PubMed - indexed for MEDLINE] 1745: Nat Biotechnol. 2002 Sep;20(9):862. US federal agencies add extra steps for handling GM plants. Fox JL. Publication Types: News PMID: 12205494 [PubMed - indexed for MEDLINE] 1746: J Biol Chem. 2002 Nov 15;277(46):44131-9. Epub 2002 Aug 29. Abrogation of upstream open reading frame-mediated translational control of a plant S-adenosylmethionine decarboxylase results in polyamine disruption and growth perturbations. Hanfrey C, Franceschetti M, Mayer MJ, Illingworth C, Michael AJ. Division of Food Safety Science, Institute of Food Research, Norwich Research Park, Colney, United Kingdom. S-Adenosylmethionine decarboxylase (AdoMetDC) is a key enzyme in polyamine biosynthesis. We show that the plant AdoMetDC activity is subject to post-transcriptional control by polyamines. A highly conserved small upstream open reading frame (uORF) in the AdoMetDC mRNA 5' leader is responsible for translational repression of a downstream beta-glucuronidase reporter cistron in transgenic tobacco plants. Elimination of the small uORF from an AdoMetDC cDNA led to increased relative translational efficiency of the AdoMetDC proenzyme in transgenic plants. The resulting increased activity of AdoMetDC caused disruption to polyamine levels with depletion of putrescine, reduction of spermine levels, and a more than 400-fold increase in the level of decarboxylated S-adenosylmethionine. These changes were associated with severe growth and developmental defects. The high level of decarboxylated S-adenosylmethionine was not associated with any change in 5'-methylcytosine content in genomic DNA and S-adenosylmethionine levels were more or less normal, indicating a highly efficient system for maintenance of S-adenosylmethionine levels in plants. This work demonstrates that uORF-mediated translational control of AdoMetDC is essential for polyamine homeostasis and for normal growth and development. Publication Types: Research Support, Non-U.S. Gov't PMID: 12205086 [PubMed - indexed for MEDLINE] 1747: Science. 2002 Aug 23;297(5585):1257. Animal biotechnology. Environmental impact seen as biggest risk. Stokstad E. Publication Types: News PMID: 12193761 [PubMed - indexed for MEDLINE] 1748: Nature. 2002 Aug 22;418(6900):805. Environmental impact tops list of fears about transgenic animals. Check E. Publication Types: News PMID: 12192374 [PubMed - indexed for MEDLINE] 1749: Clin Exp Allergy. 2002 Aug;32(8):1131-43. Report on the potential allergenicity of genetically modified organisms and their products. Lack G, Chapman M, Kalsheker N, King V, Robinson C, Venables K; BSACI working party. Department of Paediatric allergy & Immunology, St. Mary's Hospital, London, UK. gideon.lack@st-marys.nhs.uk PMID: 12190648 [PubMed - indexed for MEDLINE] 1750: Pol J Vet Sci. 2002;5(2):123-5. Contaminants in feed for food-producing animals. Moreno-López J. Department of Veterinary Microbiology, Swedish University of Agricultural Sciences (SLU), P.O. Box 585, Uppsala, 75 123 Sweden. Outbreaks of Bovine Spongiform Encephalopathy (BSE) and food borne microbial infections, dioxin contaminated animal products, the presence of veterinary drug residues, microbial resistance to antibiotics, mycotoxins, agricultural and industrial chemicals, etc. are serious concerns for the food industry in many countries. Since the direct links between feed safety and safety of foods of animal origin are obvious, feed production and manufacture should be considered as an integral part of the food production chain. Industry is responsible for the quality and safety of food and feed that is produced. This paper is a brief review of some microorganisms as source of infections for farm animals that could result in human illnesses. These include Salmonella enterica, Bacillus anthracis, Toxoplasma gondii, Trichinella spiralis, prions, Listeria monocytogenes, EHEC, Campylobacter, Clostridium botulinum, Hog Cholera virus, Foot and Mouth Disease virus, etc. as well as other contaminants associated with animal feed such as mycotoxins, veterinary drugs, dioxins and PCB and Genetically Modified Organisms. PMID: 12189948 [PubMed - indexed for MEDLINE] 1751: Plant J. 2002 Aug;31(4):477-86. Modifying the pollen coat protein composition in Brassica. Foster E, Schneiderman D, Cloutier M, Gleddie S, Robert LS. Eastern Cereal and Oilseed Research Centre, Agriculture and Agri-Food Canada, 2091 K.W. Neatby Bldg., 960 Carling Ave., Ottawa, ON K1A 0C6, Canada. The interactions between pollen and stigma are essential for plant reproduction and are made possible by compounds, such as proteins and lipids, located on their surfaces. The pollen coat is formed in part by compounds synthesized in, and released from, the tapetum, which become transferred to the pollen coat late in pollen development. In the Brassicaceae the predominant proteins of the mature pollen coat are the tapetal oleosin-like proteins, which are highly expressed in, and ultimately transferred from, the tapetum. Here we report the modification of the protein composition of the pollen coat by the addition of an active enzyme which was synthesized in the tapetum. The marker enzyme beta-glucuronidase (GUS) was successfully targeted to the pollen coat in transgenic Brassica carinata plants expressing GUS translationally fused to a B. napus tapetal oleosin-like protein (BnOlnB;4). To our knowledge this is the first demonstration of the targeting of an enzyme to the pollen coat. PMID: 12182705 [PubMed - indexed for MEDLINE] 1752: Plant J. 2002 Aug;31(4):423-30. Five years of Bt cotton in China - the benefits continue. Pray CE, Huang J, Hu R, Rozelle S. Department of Agricultural, Food and Resource Economics, Rutgers University, New Brunswick, NJ, USA. pray@aesop.rutgers.edu Bt cotton is spreading very rapidly in China, in response to demand from farmers for technology that will reduce both the cost of pesticide applications and exposure to pesticides, and will free up time for other tasks. Based on surveys of hundreds of farmers in the Yellow River cotton-growing region in northern China in 1999, 2000 and 2001, over 4 million smallholders have been able to increase yield per hectare, and reduce pesticide costs, time spent spraying dangerous pesticides, and illnesses due to pesticide poisoning. The expansion of this cost-saving technology is increasing the supply of cotton and pushing down the price, but prices are still sufficiently high for adopters of Bt cotton to make substantial gains in net income. Publication Types: Comparative Study Research Support, Non-U.S. Gov't Review PMID: 12182701 [PubMed - indexed for MEDLINE] 1753: Plant J. 2002 Aug;31(4):387-406. Insect-resistant transgenic plants in a multi-trophic context. Groot AT, Dicke M. Laboratory of Entomology, Wageningen University, PO Box 8031, 6700 EH Wageningen, The Netherlands. So far, genetic engineering of plants in the context of insect pest control has involved insertion of genes that code for toxins, and may be characterized as the incorporation of biopesticides into classical plant breeding. In the context of pesticide usage in pest control, natural enemies of herbivores have received increasing attention, because carnivorous arthropods are an important component of insect pest control. However, in plant breeding programmes, natural enemies of herbivores have largely been ignored, although there are many examples that show that plant breeding affects the effectiveness of biological control. Negative influences of modified plant characteristics on carnivorous arthropods may induce population growth of new, even more harmful pest species that had no pest status prior to the pesticide treatment. Sustainable pest management will only be possible when negative effects on non-target, beneficial arthropods are minimized. In this review, we summarize the effects of insect-resistant crops and insect-resistant transgenic crops, especially Bt crops, from a food web perspective. As food web components, we distinguish target herbivores, non-target herbivores, pollinators, parasitoids and predators. Below-ground organisms such as Collembola, nematodes and earthworms should also be included in risk assessment studies, but have received little attention. The toxins produced in Bt plants retain their toxicity when bound to the soil, so accumulation of these toxins is likely to occur. Earthworms ingest the bound toxins but are not affected by them. However, earthworms may function as intermediaries through which the toxins are passed on to other trophic levels. In studies where effects of insect-resistant (Bt) plants on natural enemies were considered, positive, negative and no effects have been found. So far, most studies have concentrated on natural enemies of target herbivores. However, Bt toxins are structurally rearranged when they bind to midgut receptors, so that they are likely to lose their toxicity inside target herbivores. What happens to the toxins in non-target herbivores, and whether these herbivores may act as intermediaries through which the toxins may be passed on to the natural enemies, remains to be studied. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 12182699 [PubMed - indexed for MEDLINE] 1754: Mol Plant Microbe Interact. 2002 Aug;15(8):753-63. Sequence-, tissue-, and delivery-specific targeting of RNA during post-transcriptional gene silencing. Balmori-Melian E, MacDiarmid RM, Beck DL, Gardner RC, Forster RL. The Horticulture and Food Research Institute of New Zealand Limited, Auckland. Transgenic Nicotiana benthamiana plants expressing an untranslatable version of the coat protein (CP) gene from the Tamarillo mosaic virus (TaMV) were either resistant to TaMV infection or recovered from infection. These phenotypes were the result of a post-transcriptional gene silencing (PTGS) mechanism that targeted TaMV-CP sequences for degradation. The TaMV-CP sequences were degraded when present in the wild-type TaMV potyvirus, in transgene mRNA, or in chimeric viral vectors based on White clover mosaic virus. The more efficiently targeted region was mapped to a 134-nt segment. Differences were observed in the efficiency of targeting during cell-to-cell and long-distance movement of the chimeric viruses. However, the TaMV-CP sequences do not appear to be targeted for degradation when delivered by biolistics. PMID: 12182332 [PubMed - indexed for MEDLINE] 1755: Exp Toxicol Pathol. 2002 Jul;54(1):57-9. Genetically modified foods: hazard identification and risk assessment. Session summary. Ito N. First Department of Pathology, Nagoya City University, Aichi, Japan. nobi-i@hkg.odn.ne.jp During a Joint Society of Toxicologic Pathology (STP)/International Federation of Societies of Toxicologic Pathologists (IFSTP) International Symposium, held between June 24 and 28, 2001, in Orlando, FL, USA, there was a session entitled as "Genetically Modified Foods: Hazard Identification and Risk Assessment". The purpose of this session was to present and discuss the current situations in the US, European Union and Japan for the public concerns, safety assessments and regulations on genetically modified (GM) products used as foods or food ingredients. Assuming the wide and fast growing of the usage of GM products, it is the duty for us as toxicologic pathologists, to supply reliable data on their safety and possible risks or hazards as a world-wide basis to not only governments or regulatory agencies but also general public of our countries. Publication Types: Congresses PMID: 12180803 [PubMed - indexed for MEDLINE] 1756: J AOAC Int. 2002 Jul-Aug;85(4):938-44. Real-time detection of genetically modified soya using Lightcycler and ABI 7700 platforms with TaqMan, Scorpion, and SYBR Green I chemistries. Terry CF, Shanahan DJ, Ballam LD, Harris N, McDowell DG, Parkes HC. Bioanalytical Innovations (F008), LGC (Teddington) Ltd., Middlesex, UK. cft@lgc.co.uk A comparative cross platform evaluation of real-time polymerase chain reaction detection of DNA sequences present in Roundup Ready soya was undertaken using the ABI 7700 and Roche Lightcycler detection systems in combination with 3 different detection chemistries: TaqMan, Scorpion primers, and SYBR Green I fluorescent dye. Various copy numbers of a plasmid containing the soya lectin sequence were used to determine the sensitivity and reproducibility of the different technology combinations and to examine both inter and intra machine variability. To examine the relative accuracy of each technology, the genetically modified soya content of baked products containing known amounts of Roundup Ready soya was determined by detection of lectin and the EPSPS transgene. It was determined that the combination of TaqMan detection chemistry and the ABI 7700 platform represented the best method for quantitative detection of genetically modified organisms in terms of both precision and accuracy. Publication Types: Research Support, Non-U.S. Gov't PMID: 12180691 [PubMed - indexed for MEDLINE] 1757: Trends Biotechnol. 2002 Sep;20(9):402-4. Agricultural biotechnology and the UK public. Moses V. CropGen, 31 St. Petersburgh Place, London, UK W2 4LL. v.moses@qmul.ac.uk It might be an exaggeration to claim that UK agriculture is in a state of crisis, but it is certainly a time for decisions. Uncertainties abound: the implications of growing free trade; a steady reduction in subsidies; the ongoing drift of people to towns and what some regard as the stranglehold of retailers both on food prices and on what the farmer gets. To all of this has to be added the backwash of bovine spongiform encephalopathy (BSE) and foot-and-mouth disease (FMD), and the advent of new technologies, especially those based on modern genetics. PMID: 12175772 [PubMed - indexed for MEDLINE] 1758: Trends Biotechnol. 2002 Sep;20(9):376-81. Biotechnology in the global agri-food system. Phillips PW. Managing Knowledge-based Agri-food Development, Agricultural Economics, University of Saskatchewan, 51 Campus Drive, Saskatoon, Canada S7N 5A8. phillips@duke.usask.ca The advent of biotechnology presents fundamental challenges to the global agri-food industry. While the scientific base for agri-food production is being revolutionised, it is not clear if or how the technology will be used. Proponents of biotechnology and a large portion of agri-food policy makers around the world project a positive future in which technology overcomes food shortages, improves the environment, heals or eliminates disease and leads to a prosperous and healthy society. A smaller but significant array of policy makers, citizens and consumers fear that the technology will exacerbate food insecurity, threaten the environment, endanger human health and ultimately impoverish society itself. Although scientists and industry are convinced the fears are unfounded, it is not clear that our social institutions will be able to adapt, adopt and use the technology in a way that will satisfy society and improve social welfare. PMID: 12175768 [PubMed - indexed for MEDLINE] 1759: Trends Biotechnol. 2002 Sep;20(9):374-5; author reply 375. India's wasteful war on biotechnology. Miller HI. Publication Types: Letter PMID: 12175766 [PubMed - indexed for MEDLINE] 1760: Physiol Behav. 2002 May 1;76(1):21-6. Food-anticipatory activity and liver per1-luc activity in diabetic transgenic rats. Davidson AJ, Stokkan KA, Yamazaki S, Menaker M. Collaborators: Menaker M. Department of Biology, University of Virginia, Charlottesville, VA 22904, USA. The mammalian Per1 gene is an important component of the core cellular clock mechanism responsible for circadian rhythms. The rodent liver and other tissues rhythmically express Per1 in vitro but typically damp out within a few cycles. In the liver, the peak of this rhythm occurs in the late subjective night in an ad lib-fed rat, but will show a large phase advance in response to restricted availability of food during the day. The relationship between this shift in the liver clock and food-anticipatory activity (FAA), the circadian behavior entrained by daily feeding, is currently unknown. Insulin is released during feeding in mammals and could serve as an entraining signal to the liver. To test the role of insulin in the shift in liver Per1 expression and the generation of FAA, per-luciferase transgenic rats were made diabetic with a single injection of streptozotocine. Following 1 week of restricted feeding and locomotor activity monitoring, liver was collected for per-luc recording. In two separate experiments, FAA emerged and liver Per1 phase-shifted in response to daytime 8-h food restriction. The results rule out insulin as a necessary component of this system. Publication Types: Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S. PMID: 12175585 [PubMed - indexed for MEDLINE] 1761: Plant Cell. 2002 Aug;14(8):1767-85. Discrete forms of amylose are synthesized by isoforms of GBSSI in pea. Edwards A, Vincken JP, Suurs LC, Visser RG, Zeeman S, Smith A, Martin C. John Innes Centre, Colney, Norwich NR4 7UH, United Kingdom. Amyloses with distinct molecular masses are found in the starch of pea embryos compared with the starch of pea leaves. In pea embryos, a granule-bound starch synthase protein (GBSSIa) is required for the synthesis of a significant portion of the amylose. However, this protein seems to be insignificant in the synthesis of amylose in pea leaves. cDNA clones encoding a second isoform of GBSSI, GBSSIb, have been isolated from pea leaves. Comparison of GBSSIa and GBSSIb activities shows them to have distinct properties. These differences have been confirmed by the expression of GBSSIa and GBSSIb in the amylose-free mutant of potato. GBSSIa and GBSSIb make distinct forms of amylose that differ in their molecular mass. These differences in product specificity, coupled with differences in the tissues in which GBSSIa and GBSSIb are most active, explain the distinct forms of amylose found in different tissues of pea. The shorter form of amylose formed by GBSSIa confers less susceptibility to the retrogradation of starch pastes than the amylose formed by GBSSIb. The product specificity of GBSSIa could provide beneficial attributes to starches for food and nonfood uses. Publication Types: Comparative Study Research Support, Non-U.S. Gov't PMID: 12172021 [PubMed - indexed for MEDLINE] 1762: Pathol Biol (Paris). 2002 Jul;50(6):380-7. [Transgenes and their medical applications] [Article in French] Houdebine LM. Unité de biologie du développement et biotechnologie, institut national de la recherche agronomique, 78352 Jouy-en-Josas, France. houdebine@jouy.inra.fr Transgenesis consists of introducing stably a foreign genetic information into the genome of a multicellular organism. These techniques used for the first time in 1980 for the animals and 1983 for plants have multiple applications of which many are directly or not related to medicine. Transgenesis has become one of the essential tools to study the role of genes in the control of biological functions. This approach is logically accompanied by the generation of transgenic animal lines for the study of human diseases and the test of new pharmaceuticals. Milk from transgenic animals as well as leaves and seeds from transgenic plants are ready to become an essential source of proteins having essential therapeutical effects. Genetically modified pigs are expected to be resistant to rejection mechanisms to become the source of organs or cells for patients. The application of transgenesis in agriculture and breeding is still in infancy. It may contribute quite significantly to provide human communities with food in sufficient amount, having improved nutritional properties and produced using a milder and less polluting methods. Publication Types: English Abstract Review PMID: 12168256 [PubMed - indexed for MEDLINE] 1763: Nature. 2002 Aug 8;418(6898):696-9. The present and future of the international wine industry. Bisson LF, Waterhouse AL, Ebeler SE, Walker MA, Lapsley JT. Department of Viticulture and Enology, University of California, Davis, One Shields Avenue, Davis, California 95616-8749, USA. Wine production is both art and science, a blend of individual creativity and innovative technology. But wine production is also business, with economic factors driving manufacturing practices. To be successful in the modern marketplace, a winemaker must integrate the artistic and economic aspects of wine production, and possess a solid understanding of the intrinsic and extrinsic factors that underlie purchase motivation. PMID: 12167877 [PubMed - indexed for MEDLINE] 1764: Nature. 2002 Aug 8;418(6898):685-8. Assessing the risks associated with new agricultural practices. Hails RS. NERC Centre for Ecology and Hydrology, Oxford, UK. rha@ceh.ac.uk One key challenge for the twenty-first century is how to produce the food we need, yet ensure the landscape we want. Genetically modified crops have focused our attention on how to answer this question for one part of agriculture. The same principles could be applied to assess environmental impacts of future land-use change in a much broader context. PMID: 12167875 [PubMed - indexed for MEDLINE] 1765: Nature. 2002 Aug 8;418(6898):678-84. Enhancing the crops to feed the poor. Huang J, Pray C, Rozelle S. Center for Chinese Agricultural Policy, Institute of Geographic Sciences and Natural Resource Research, Chinese Academy of Sciences, 917 Building, Anwai Datun Road, Beijing 100101, China. Solutions to the problem of how the developing world will meet its future food needs are broader than producing more food, although the successes of the 'Green Revolution' demonstrate the importance of technology in generating the growth in food output in the past. Despite these successes, the world still faces continuing vulnerability to food shortages. Given the necessary funding, it seems likely that conventional crop breeding, as well as emerging technologies based on molecular biology, genetic engineering and natural resource management, will continue to improve productivity in the coming decades. Publication Types: Review PMID: 12167874 [PubMed - indexed for MEDLINE] 1766: Nature. 2002 Aug 8;418(6898):668-70. Comment in: Nature. 2002 Oct 24;419(6909):777. Malthus foiled again and again. Trewavas A. Institute of Cell and Molecular Biology, University of Edinburgh, UK. trewavas@ed.ac.uk Throughout history, increasing population has driven the need to increase agricultural efficiency, so averting successive 'malthusian' disasters. In the twentieth century, the application of scientific knowledge to agriculture yielded tremendous dividends, enabling cereal yields to increase threefold since 1950. But with the world's population projected to reach nine billion by the middle of this century, new ways must be found to increase yields while preserving natural habitats and biodiversity. Publication Types: Historical Article PMID: 12167872 [PubMed - indexed for MEDLINE] 1767: Nature. 2002 Aug 8;418(6898):571. US calls for early data on transgenic crop safety. Check E. Publication Types: News PMID: 12167819 [PubMed - indexed for MEDLINE] 1768: Nature. 2002 Aug 8;418(6898):571-2. Comment in: Nature. 2002 Aug 8;418(6898):569. Africa hungry for conventional food as biotech row drags on. McDowell N. Publication Types: News PMID: 12167818 [PubMed - indexed for MEDLINE] 1769: Nature. 2002 Aug 8;418(6898):569. Comment on: Nature. 2002 Aug 8;418(6898):571-2. Poverty and transgenic crops. [No authors listed] Publication Types: Comment Editorial PMID: 12167817 [PubMed - indexed for MEDLINE] 1770: Trends Plant Sci. 2002 Aug;7(8):366-73. Strategies for transgenic manipulation of monoterpene biosynthesis in plants. Mahmoud SS, Croteau RB. Institute of Biological Chemistry, Washington State University, Pullman, WA 99164, USA. Monoterpenes, the C(10) isoprenoids, are a large family of natural products that are best known as constituents of the essential oils and defensive oleoresins of aromatic plants. In addition to ecological roles in pollinator attraction, allelopathy and plant defense, monoterpenes are used extensively in the food, cosmetic and pharmaceutical industries. The importance of these plant products has prompted the definition of many monoterpene biosynthetic pathways, the cloning of the relevant genes and the development of genetic transformation techniques for agronomically significant monoterpene-producing plants. Metabolic engineering of monoterpene biosynthesis in the model plant peppermint has resulted in yield increase and compositional improvement of the essential oil, and also provided strategies for manipulating flavor and fragrance production, and plant defense. Publication Types: Review PMID: 12167332 [PubMed - indexed for MEDLINE] 1771: Vaccine. 2002 Aug 19;20(25-26):3155-64. Expression in plants and immunogenicity of plant virus-based experimental rabies vaccine. Yusibov V, Hooper DC, Spitsin SV, Fleysh N, Kean RB, Mikheeva T, Deka D, Karasev A, Cox S, Randall J, Koprowski H. Biotechnology Foundation Laboratories at Thomas Jefferson University, 1020 Locust Street, Room 346 JAH, Philadelphia, PA 19107, USA. A new approach to the production and delivery of vaccine antigens is the use of engineered amino virus-based vectors. A chimeric peptide containing antigenic determinants from rabies virus glycoprotein (G protein) (amino acids 253-275) and nucleoprotein (N protein) (amino acids 404-418) was PCR-amplified and cloned as a translational fusion product with the alfalfa mosaic virus (AlMV) coat protein (CP). This recombinant CP was expressed in two plant virus-based expression systems. The first one utilized transgenic Nicotiana tabacum cv. Samsun NN plants providing replicative functions in trans for full-length infectious RNA3 of AlMV (NF1-g24). The second one utilized Nicotiana benthamiana and spinach (Spinacia oleracea) plants using autonomously replicating tobacco mosaic virus (TMV) lacking native CP (Av/A4-g24). Recombinant virus containing the chimeric rabies virus epitope was isolated from infected transgenic N. tabacum cv. Samsun NN plants and used for parenteral immunization of mice. Mice immunized with recombinant virus were protected against challenge infection. Based on the previously demonstrated efficacy of this plant virus-based experimental rabies vaccine when orally administered to mice in virus-infected unprocessed raw spinach leaves, we assessed its efficacy in human volunteers. Three of five volunteers who had previously been immunized against rabies virus with a conventional vaccine specifically responded against the peptide antigen after ingesting spinach leaves infected with the recombinant virus. When rabies virus non-immune individuals were fed the same material, 5/9 demonstrated significant antibody responses to either rabies virus or AlMV. Following a single dose of conventional rabies virus vaccine, three of these individuals showed detectable levels of rabies virus-neutralizing antibodies, whereas none of five controls revealed these antibodies. These findings provide clear indication of the potential of the plant virus-based expression systems as supplementary oral booster for rabies vaccinations. Copyright 2002 Elsevier Science Ltd. Publication Types: Evaluation Studies Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. PMID: 12163267 [PubMed - indexed for MEDLINE] 1772: Appetite. 2002 Aug;39(1):9-24. Consumer attitudes towards genetically modified foods. Magnusson MK, Koivisto Hursti UK. Department of Public Health and Caring Sciences, Section of Caring Sciences, University of Uppsala, Uppsala Science Park, S-751 83 Uppsala, Sweden. maria.magnusson@pubcare.uu.se The present study reports attitudes towards genetically modified (GM) foods among Swedish consumers. A random nation-wide sample of 2,000 addressees, aged 18-65 years, were mailed a questionnaire and 786 (39%) responded. Most of these consumers were rather negative about GM foods. However, males, younger respondents and those with higher level of education were more positive than were females, older respondents and those with lower level of education. A majority of the consumers had moral and ethical doubts about eating GM foods and did not perceive attributes like better taste or lower price beneficial enough to persuade them to purchase GM foods. However, tangible benefits, like being better for the environment or healthier, seemed to increase willingness to purchase GM foods. Publication Types: Research Support, Non-U.S. Gov't PMID: 12160561 [PubMed - indexed for MEDLINE] 1773: Nat Biotechnol. 2002 Aug;20(8):775-6. Comment on: Nat Biotechnol. 2002 Jun;20(6):537-41. Terminator no solution to gene flow. Shand H. Publication Types: Comment Letter PMID: 12147996 [PubMed - indexed for MEDLINE] 1774: Nat Biotechnol. 2002 Aug;20(8):758-9. Political will to lift the GMO moratorium emerging in Europe. Meldolesi A. Publication Types: News PMID: 12147986 [PubMed - indexed for MEDLINE] 1775: Nat Biotechnol. 2002 Aug;20(8):756-7. European Parliament vote encourages industry to proclaim green biotech. Hodgson J. Publication Types: News PMID: 12147985 [PubMed - indexed for MEDLINE] 1776: Nat Biotechnol. 2002 Aug;20(8):753. Bogged down in CAP reform. [No authors listed] Publication Types: Editorial PMID: 12147983 [PubMed - indexed for MEDLINE] 1777: Nat Biotechnol. 2002 Aug;20(8):753. Caution: may be harmless if swallowed. [No authors listed] Publication Types: Editorial PMID: 12147982 [PubMed - indexed for MEDLINE] 1778: Trends Biotechnol. 2002 Aug;20(8):338-43. Biological substitutes for pesticides. Gerhardson B. Plant Pathology & Biocontrol Unit, P.O. Box 7035, S-750 07 Uppsala, Sweden. Berndt.Gerhardson@vpat.slu.se In the 20th century an increasing number of pesticides, based on biocidal molecules, were the means for a substantial increase in food and fibre production and quality. Because of health and environmental concerns continued extensive use of such molecules is intensively debated and substitutes are often urgently required. Beside crop plant resistance, various biological control methods based on natural pest suppressing organisms are regarded as main alternatives. Several approaches and concepts also have been tested and commercial organism-based preparations are steadily increasing. However, further biotechnological efforts are required to give them status of being practical substitutes to pesticides. At present they are not comparable to pesticides in meeting efficacy, market and other expectations, but they still have a promising future, especially where genetically modified organisms can be used. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 12127281 [PubMed - indexed for MEDLINE] 1779: J Biotechnol. 2002 Sep 11;98(1):107-12. The application of biotechnological methods in authenticity testing. Popping B. Eurofins Scientific, 69a Kilnwick Road, Pocklington, YO42 2JY, UK. bertpopping@eurofins.com By counterfeiting brand names in the food and drink industry as well as fraudulently labelling and selling low quality products as premium products, this sector of the industry has lost significant amounts of money and the consumer has been deceived. While it was difficult to establish certain types of fraud before the advent of modern biotechnology, DNA-based methods make an important contribution to protect high-quality brand names and protect the consumer. Several years ago, DNA technologies were considered as methods used in universities, primarily for research purpose, not so much for 'real-life' applications. However, this has changed and a number of laboratories have specialised in offering such services to the industry. This article will review DNA-based techniques commonly used for authenticity testing. PMID: 12126809 [PubMed - indexed for MEDLINE] 1780: J Biotechnol. 2002 Sep 11;98(1):79-106. Assuring the safety of genetically modified (GM) foods: the importance of an holistic, integrative approach. Cockburn A. Monsanto UK Ltd, The Maris Centre, 45 Hauxton Road, Trumpington, Cambridge CB2 2LQ, UK. andrew.cockburn@monsanto.com Genes change continuously by natural mutation and recombination enabling man to select and breed crops having the most desirable traits such as yield or flavour. Genetic modification (GM) is a recent development which allows specific genes to be identified, isolated, copied and inserted into other plants with a high level of specificity. The food safety considerations for GM crops are basically the same as those arising from conventionally bred crops, very few of which have been subject to any testing yet are generally regarded as being safe to eat. In contrast a rigorous safety testing paradigm has been developed for GM crops, which utilises a systematic, stepwise and holistic approach. The resultant science based process, focuses on a classical evaluation of the toxic potential of the introduced novel trait and the wholesomeness of the transformed crop. In addition, detailed consideration is given to the history and safe use of the parent crop as well as that of the gene donor. The overall safety evaluation is conducted under the concept known as substantial equivalence which is enshrined in all international crop biotechnology guidelines. This provides the framework for a comparative approach to identify the similarities and differences between the GM product and its comparator which has a known history of safe use. By building a detailed profile on each step in the transformation process, from parent to new crop, and by thoroughly evaluating the significance from a safety perspective, of any differences that may be detected, a very comprehensive matrix of information is constructed which enables the conclusion as to whether the GM crop, derived food or feed is as safe as its traditional counterpart. Using this approach in the evaluation of more than 50 GM crops which have been approved worldwide, the conclusion has been that foods and feeds derived from genetically modified crops are as safe and nutritious as those derived from traditional crops. The lack of any adverse effects resulting from the production and consumption of GM crops grown on more than 300 million cumulative acres over the last 5 years supports these safety conclusions. PMID: 12126808 [PubMed - indexed for MEDLINE] 1781: J Biotechnol. 2002 Sep 11;98(1):9-24. Attitudes toward biotechnology in the European Union. Pardo R, Midden C, Miller JD. Fundacion BBVA, Principe de Vergara 51, 28006, Madrid, Spain. rpardo@fbbva.es Public attitudes toward biotechnology in the European Union have been characterized as negative using Eurobarometer data, but so far little attention has been paid to building a robust metric appropriate for emerging public opinion issues which combine high salience with very limited knowledge by the public. On the basis of the general literature about the formation and structure of attitudes and about public perceptions of science, this article presents a new metric and analysis: first, for estimating the level of awareness and knowledge of biotechnology in Europe; second, for assessing the stability and depth of these evaluative perceptions; and third, for exploring the roles of canonical socio-demographic variables, the knowledge variable and general attitudinal schemas for understanding the perceptions of both benefits and risks of biotech applications. The results show the importance of general value orientations or "worldviews" in shaping positive attitudes, and more of these general cognitive schemas should be measured in future research. The same multivariate model was unable to account for a significant percentage of the total variance in the perception of risks, suggesting that new measures are needed to tap this critical area in the acceptance of biotech in Europe. PMID: 12126802 [PubMed - indexed for MEDLINE] 1782: J Biotechnol. 2002 Sep 11;98(1):3-8. People's concerns about biotechnology: some problems and some solutions. Braun R. BIOLINK, Enggisteinstrasse 19, CH 3076, Worb, Switzerland. rdbraun@bluewin.ch Pharmaceuticals and vaccines made by genetic engineering are well accepted all over the world. In contrast, there are many people, particularly in Europe, who are worried that food, made by the same new technology, may harm their health or cause damage to the environment. This is despite the growing evidence that genetically modified crops have the potential to improve world food security and the fact that there have, as yet, been no adverse results of their use in the food chain. Because of these worries and the mechanisms of politics, agricultural biotechnology has become the target of concerns about food safety (BSE, Foot & Mouth Disease), along with globalisation and the power of multinational companies. These concerns will, hopefully, be overcome by a more open and well-informed dialogue between scientists, opinion leaders, educators and the public. If judiciously applied, genetically modified crops will help increase sustainability and the fight against hunger in the world. PMID: 12126801 [PubMed - indexed for MEDLINE] 1783: Am J Med Sci. 2002 Jul;324(1):14-30. Therapeutic applications of monoclonal antibodies. Berger M, Shankar V, Vafai A. Emory University School of Public Health, Atlanta, Georgia, USA. Researchers have sought therapeutic applications for monoclonal antibodies since their development in 1975. However, murine-derived monoclonal antibodies may cause an immunogenic response in human patients, reducing their therapeutic efficacy. Chimeric and humanized antibodies have been developed that are less likely to provoke an immune reaction in human patients than are murine-derived antibodies. Antibody fragments, bispecific antibodies, and antibodies produced through the use of phage display systems and genetically modified plants and animals may aid researchers in developing new uses for monoclonal antibodies in the treatment of disease. Monoclonal antibodies may have a number of promising potential therapeutic applications in the treatment of asthma, autoimmune diseases, cancer, poisoning, septicemia, substance abuse, viral infections, and other diseases. Publication Types: Review PMID: 12120821 [PubMed - indexed for MEDLINE] 1784: Plant Cell. 2002 Jul;14(7):1497-508. A surveillance system regulates selective entry of RNA into the shoot apex. Foster TM, Lough TJ, Emerson SJ, Lee RH, Bowman JL, Forster RL, Lucas WJ. Horticulture and Food Research Institute of New Zealand, Tennent Drive, Private Bag 11030, Palmerston North, New Zealand. Phloem-mobile endogenous RNA is trafficked selectively into the shoot apex. In contrast, most viruses and long-distance post-transcriptional gene silencing (PTGS) signals are excluded from the shoot apex. These observations suggest the operation of an underlying regulatory mechanism. To examine this possibility, a potexvirus movement protein, known to modify cell-to-cell trafficking and PTGS, was expressed ectopically in transgenic plants. These plants were found to be compromised in their capacity to exclude both viral RNA and silencing signals from the shoot apex. The transgenic plants also displayed various degrees of abnormal leaf polarity depending on transgene expression level. Normal patterns of organ development were restored by either virus- or Agrobacterium tumefaciens-mediated induction of PTGS. This revealed the presence of an RNA signal surveillance system that acts to allow the selective entry of RNA into the shoot apex. We propose that this surveillance system regulates signaling and protects the shoot apex, in particular the cells that give rise to reproductive structures, from viral invasion. Publication Types: Research Support, Non-U.S. Gov't PMID: 12119370 [PubMed - indexed for MEDLINE] 1785: Trends Plant Sci. 2002 Jul;7(7):309-15. A long way ahead: understanding and engineering plant metal accumulation. Clemens S, Palmgren MG, Krämer U. Leibniz Institute of Plant Biochemistry, Weinberg 3, D-06120 Halle, Germany. sclemens@ipb-halle.de Some plants can hyperaccumulate metal ions that are toxic to virtually all other organisms at low dosages. This trait could be used to clean up metal-contaminated soils. Moreover, the accumulation of heavy metals by plants determines both the micronutrient content and the toxic metal content of our food. Complex interactions of transport and chelating activities control the rates of metal uptake and storage. In recent years, several key steps have been identified at the molecular level, enabling us to initiate transgenic approaches to engineer the transition metal content of plants. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 12119168 [PubMed - indexed for MEDLINE] 1786: Transgenic Res. 2002 Jun;11(3):249-56. Expression of an antisense GIGANTEA (GI) gene fragment in transgenic radish causes delayed bolting and flowering. Curtis IS, Nam HG, Yun JY, Seo KH. Department of Life Science, Pohang University of Science and Technology, Kyungbuk, Republic of Korea. iscurtis@postman.riken.go.jp A late-flowering transgenic radish has been produced by the expression of an antisense GIGANTEA (GI) gene fragment using a floral-dip method. Twenty-five plants were dipped into a suspension of Agrobacterium carrying a 2.5 kb antisense GI gene fragment from Arabidopsis, along with the gusA and bar reporter genes, all under the control of a CaMV 35S promoter. From a total of 1462 seeds harvested from these floral-dipped plants, 16 Basta-resistant T1 plants were found to have GUS activity (transformation efficiency of 1.1%). Southern analysis confirmed the integration of one or two copies of the gusA gene in these herbicide-resistant plants. Expression of the GI gene in T1 plants was much reduced compared to both wildtype plants and plants transformed with pCAMBIA3301 (positive control). In the progenies of eleven T1 plants analysed (T2 generation), all lines showed a significant delay in both bolting and flowering times compared to wildtype and positive control plants, and that, the level of GI transcript was inversely proportional to the time of bolting and flowering. At a maximum, bolting and flowering times were delayed by 17 and 18 days respectively, compared to wildtype plants (in positive control plants, the delay was 23 and 26 days, respectively). Ten of the 11 lines exhibited a significant reduction in plant height compared to wildtype and positive control plants. This study provides evidence that down-regulation of the GI gene by co-suppression could delay bolting in a cold-sensitive long-day (LD) plant. Production of late-flowering germplasms of radish may allow this important crop to be cultivated over an extended period and also provide further food to the famine countries of S/E Asia. Publication Types: Research Support, Non-U.S. Gov't PMID: 12113457 [PubMed - indexed for MEDLINE] 1787: Nature. 2002 Jul 11;418(6894):114. Comment in: Nature. 2002 Sep 26;419(6905):337. Europe gets tough on labelling genetically modified foodstuffs. Butler D. Publication Types: News PMID: 12110851 [PubMed - indexed for MEDLINE] 1788: Nahrung. 2002 Jun;46(3):179-83. Quantitative and qualitative analysis of lipids in genetically modified potato tubers with varying rates of 14-3-3 protein synthesis. Prescha A, Biernat J, Szopa J. Department of Food Science and Nutrition, Wrocław Medical University, Poland. In six transgenic lines of potatoes with the varying rates of 14-3-3 protein synthesis as well as in control cultivar Desiree the content and composition of the lipids extracted from the mature tubers from three years field trials (1998-2000) were analyzed. The transgenic lines J2 and J1 are both overexpressing gene encoding 14-3-3 protein. The J2 exhibited an overexpression of the protein 14-3-3 derived from pumpkin (Cucurbita pepo) cDNA and in J1 the 14-3-3 overexpression resulted from modifying of ADP-ribosylation factor synthesis. In the remaining lines, synthesis of the protein 14-3-3 was modified by the antisense technology. In tubers from 1998, the content of total lipids was within the range of 0.45-0.88% of tuber dry matter. The highest amount of fat was in tubers of line J2 (69% more than in the control). The content of lipids in tubers from subsequent years ranged from 0.36 to 0.63% of dry matter. Consistently the highest amount of fat was in tubers of line J2, however, the increase was very slight (8.6% more than in the control). The fractionation of lipids into polar and nonpolar fractions showed that all transgenic lines from field trials 1998 and 2000 contained more nonpolar lipids than the control (up to 270% in line J2). The percentage of nonpolar fractions in fats of tubers from all transgenes harvested in 1999 were similar, but they were higher than in tubers from the previous years, and they amounted to 44.4-49.1%. Chromatographic separation of methyl esters of fatty acids demonstrated that cis-alpha-linoleic acid was the main fatty acid present in potato tubers. This acid composed the biggest part of all lipids in G2 line. In the nonpolar fraction of lipids, palmitic acid followed by cis-alpha-linoleic acid showed the highest amounts. Publication Types: Research Support, Non-U.S. Gov't PMID: 12108217 [PubMed - indexed for MEDLINE] 1789: Nutr Health. 2002;16(2):73-84. Can science give us the tools for recognizing possible health risks of GM food? Pusztai A. The Rowett Research Institute, Aberdeen, Scotland, UK. a.pusztai@freenet.co.uk Nearly ten years after the introduction of GM foodcrops there are still only a handful of published studies about their safety. Independent studies are even fewer, moreover, no peer-reviewed publications exist in which the results of clinical investigations on the possible effects of GM food on human health are described. Even though the evaluation of the safety or possible toxicity of GM foodstuffs is more difficult than that of drugs or food additives, this scarcity of data and the lack of a scientific database is curious particularly as descriptions of the results of chemical, nutritional and biological testing in some early (unpublished) studies or some more recent publications demonstrate the feasibility of carrying out proper and scientifically valid health risk assessment on GM foods. In this review, after critically examining some of the basic principles, past results and possible novel methods of future health safety assessment of GM foodstuffs, the conclusion appears to be that as the tools for the recognition and indeed for the elimination of the risks GM foods may present for us are available or can be developed, it is the will and the funding for such work that needs to be found. Publication Types: Review PMID: 12102369 [PubMed - indexed for MEDLINE] 1790: Shokuhin Eiseigaku Zasshi. 2002 Apr;43(2):68-73. Increased digestibility of two products in genetically modified food (CP4-EPSPS and Cry1Ab) after preheating. Okunuki H, Teshima R, Shigeta T, Sakushima J, Akiyama H, Goda Y, Toyoda M, Sawada J. National Institute of Health Sciences: 1-18-1, Kamiyoga, Setagaya-ku, Tokyo 158-8501, Japan. We performed experiments on in vitro digestion of newly expressed proteins by SGF (simulated gastric fluid) and SIF (simulated intestinal fluid) to assess the allergenicity of food components derived from biotechnological modification. For newly expressed proteins, we chose CP4-EPSPS (5-enolpyruvylshikimate-3-phosphate synthase from Agrobacterium sp. strain CP4) and Cry1Ab derived from Bacillus thuringiensis subsp. kurstaki strain HD-1. The former is expressed in GM-soybeans and the latter is expressed in GM-corns. Firstly, we examined the digestibility of purified CP4-EPSPS and Cry1Ab by SGF. Both proteins were rapidly digested within 60 sec. After preheating, the digestibility by SGF was slightly increased. Secondly, CP4-EPSPS in GM-soybean extracts and Cry1Ab in GM-corn extracts were digested by SGF. The digestion time of both proteins by SGF was almost the same as that of the purified proteins. Thirdly, the digestibility of CP4-EPSPS and Cry1Ab by SIF was examined. The digestion time of these proteins was 240 min or more. However, digestibility of these proteins by SIF was dramatically increased by preheating, and the digestion time was less than 5 sec. Fourthly, CP4-EPSPS in GM-soybean extracts and Cry1Ab in GM-corn extracts were digested by SIF. Digestion time of both proteins by SIF was almost the same as that of the purified proteins. From these results, we concluded that the digestibility of both CP4-EPSPS and Cry1Ab by SGF and SIF was increased by preheating. Therefore, we suggest that the allergenicity of both proteins should be extremely low because of the easy digestibility of these proteins by SGF and also by SIF with preheating. Publication Types: Research Support, Non-U.S. Gov't PMID: 12092415 [PubMed - indexed for MEDLINE] 1791: Duodecim. 2001;117(1):112-6. [Genetically modified and other novel food products] [Article in Finnish] Mannonen L. Elintarvikevirasto PL 5, 00531 Helsinki. Publication Types: Review PMID: 12092344 [PubMed - indexed for MEDLINE] 1792: Curr Drug Metab. 2002 Jun;3(3):311-9. Mutagenesis by environmental pollutants and bio-monitoring of environmental mutagens. Sato H, Aoki Y. Research Center for Environmental Risk, National Institute for Environmental Studies, Tsukuba, Japan. ybaoki@nies.go.jp There is serious concern about the adverse effects of environmental pollutants on human health. Various mutagens, which pollute air, water, and food, possibly induce mutations in humans, and are suspected of causing cancer. Environmental mutagens, such as polycyclic aromatic hydrocarbons (PAH) and heterocyclic amines are known to bind to nucleotides, resulting in the formation of DNA adducts. Some DNA adducts are fixed as mutations through replication of DNA. Reactive oxygen species generated by pollutants also induce the formation of DNA adducts. DNA adducts have been detected as a marker for the exposure of humans and wild life to mutagens. Because of its high sensitivity the 32P-postlabel-thin layer chromatography (TLC) method is widely used for the analysis of DNA adducts formed by PAH and related bulky compounds. However, new systems are required for detecting mutations induced in genomic DNA in vivo to monitor environmental mutagens. Recently, transgenic animals, in which a target gene for detecting mutations is integrated, have been developed. With these transgenic animals, not only mutant frequency but mutation spectra can be determined. We review here recent advances in the detection of DNA adducts formed by environmental pollutants and their application for biological monitoring of environmental mutagens. We also discuss transgenic animals as important tools for evaluating the total mutagenic potential of environmental chemicals. Publication Types: Review PMID: 12083323 [PubMed - indexed for MEDLINE] 1793: J AOAC Int. 2002 May-Jun;85(3):809-15. Validation studies and proficiency testing. Ankilam E, Heinze P, Kay S, Van den Eede G, Popping B. European Commission, Joint Research Centre, Food Products Unit, Ispra, Italy. elke.anklam@jrc.it Genetically modified organisms (GMOs) entered the European food market in 1996. Current legislation demands the labeling of food products if they contain <1% GMO, as assessed for each ingredient of the product. To create confidence in the testing methods and to complement enforcement requirements, there is an urgent need for internationally validated methods, which could serve as reference methods. To date, several methods have been submitted to validation trials at an international level; approaches now exist that can be used in different circumstances and for different food matrixes. Moreover, the requirement for the formal validation of methods is clearly accepted; several national and international bodies are active in organizing studies. Further validation studies, especially on the quantitative polymerase chain reaction methods, need to be performed to cover the rising demand for new extraction methods and other background matrixes, as well as for novel GMO constructs. Publication Types: Validation Studies PMID: 12083280 [PubMed - indexed for MEDLINE] 1794: J AOAC Int. 2002 May-Jun;85(3):801-8. Detection methods and performance criteria for genetically modified organisms. Bertheau Y, Diolez A, Kobilinsky A, Magin K. Institut National de la Recherche Agronomique, PMDV/MDO, Versailles, France. bertheau@versailles.inra.fr Detection methods for genetically modified organisms (GMOs) are necessary for many applications, from seed purity assessment to compliance of food labeling in several countries. Numerous analytical methods are currently used or under development to support these needs. The currently used methods are bioassays and protein- and DNA-based detection protocols. To avoid discrepancy of results between such largely different methods and, for instance, the potential resulting legal actions, compatibility of the methods is urgently needed. Performance criteria of methods allow evaluation against a common standard. The more-common performance criteria for detection methods are precision, accuracy, sensitivity, and specificity, which together specifically address other terms used to describe the performance of a method, such as applicability, selectivity, calibration, trueness, precision, recovery, operating range, limit of quantitation, limit of detection, and ruggedness. Performance criteria should provide objective tools to accept or reject specific methods, to validate them, to ensure compatibility between validated methods, and be used on a routine basis to reject data outside an acceptable range of variability. When selecting a method of detection, it is also important to consider its applicability, its field of applications, and its limitations, by including factors such as its ability to detect the target analyte in a given matrix, the duration of the analyses, its cost effectiveness, and the necessary sample sizes for testing. Thus, the current GMO detection methods should be evaluated against a common set of performance criteria. PMID: 12083279 [PubMed - indexed for MEDLINE] 1795: J AOAC Int. 2002 May-Jun;85(3):797-800. DNA methods: critical review of innovative approaches. Kok EJ, Aarts HJ, Van Hoef AM, Kuiper HA. State Institute for Quality Control of Agricultural Products (RIKILT), Department of Food Safety and Health, Wageningen, The Netherlands. e.j.kok@rikilt.wag-ur.nl The presence of ingredients derived from genetically modified organisms (GMOs) in food products in the market place is subject to a number of European regulations that stipulate which product consisting of or containing GMO-derived ingredients should be labeled as such. In order to maintain these labeling requirements, a variety of different GMO detection methods have been developed to screen for either the presence of DNA or protein derived from (approved) GM varieties. Recent incidents where unapproved GM varieties entered the European market show that more powerful GMO detection and identification methods will be needed to maintain European labeling requirements in an adequate, efficient, and cost-effective way. This report discusses the current state-of-the-art as well as future developments in GMO detection. Publication Types: Research Support, Non-U.S. Gov't PMID: 12083278 [PubMed - indexed for MEDLINE] 1796: J AOAC Int. 2002 May-Jun;85(3):792-6. State of the art and limitations of quantitative polymerase chain reaction. Wiseman G. RHM Technology Ltd, High Wycombe, Bucks, UK. gwiseman@rhmtech.co.uk Consequential to the implementation of European Commission (EC) Regulation 1139/98, EC Regulation 49/2000, and EC Regulation 50/2000 has been the need to measure accurately the levels of the genetically modified (GM) species Roundup Ready Soya and Bt 176 Maize that are present in food. Analytical methods to detect and quantitate these transgenic species have received much attention particularly with respect to the deminimus threshold of 1% for their presence in materials derived from non-GM identity-preserved (IP) supplies. The relative advantages and limitations of threshold analysis by double-competitive polymerase chain reaction (PCR) and quantitative real-time PCR are discussed in their application to the quantitative analysis of processed foods. Consideration is also given to other factors involved in the analyses that affect the performance of quantitative procedures, and to the many uncertainties involved in the precision of a reported analytical result. PMID: 12083277 [PubMed - indexed for MEDLINE] 1797: J AOAC Int. 2002 May-Jun;85(3):787-91. Detection of genetically modified organisms in foods by protein- and DNA-based techniques: bridging the methods. van Duijn GJ, van Biert R, Bleeker-Marcelis H, Van Boeijen I, Adan AJ, Jhakrie S, Hessing M. TNO Nutrition and Food Research, AJ Zeist, The Netherlands. vanDuijn@voeding.tno.nl According to European Commission (EC) Regulation 1139/98, foods and food ingredients that are to be delivered to the final consumer in which either protein or DNA resulting from genetic modification is present, shall be subject to additional specific labeling requirements. Since 1994, genetically altered tomatoes, squash, potatoes, canola, cotton, and soy have been on the market. Recently, insect-resistant and herbicide-tolerant maize varieties have been introduced. Soy and maize are 2 of the most important vegetable crops in the world. During the past 4 years, both protein- and DNA-based methods have been developed and applied for detection of transgenic soy and maize, and their derivatives. For protein-based detection, specific monoclonal and polyclonal antibodies have been developed; for immunochemical detection, Western blot analysis and enzyme-linked immunosorbent assays are the most prominent examples. For detection of genetically modified organisms (GMOs) at the level of DNA, polymerase chain reaction-based methods are mainly used. For these reactions, highly specific primer sets are needed. This study compares the principally different methods. Specificity of methods and the possible risks of false-positive or false-negative results are considered in relation to sampling, matrix effects, and food processing procedures. In addition, quantitative aspects of protein- and DNA-based GM detection methods are presented and discussed. This is especially relevant as EC regulation 49/2000, which defines a threshold for an unintentional comingling of 1%, came into force on April 10, 2000. PMID: 12083276 [PubMed - indexed for MEDLINE] 1798: J AOAC Int. 2002 May-Jun;85(3):780-6. Protein immunoassay methods for detection of biotech crops: applications, limitations, and practical considerations. Stave JW. Strategic Diagnostics, Inc, Newark, DE 19713, USA. jstave@sdix.com Immunoassay methods are available for detection and quantitation of proteins expressed by most biotechnology-derived crops in commercial production. The 2 most common test formats are enzyme-linked immunosorbent assay (ELISA) and immunochromatographic (lateral flow) strip tests. Two ELISA methods, one for Roundup Ready soybeans and one for MON810 CrylAb corn, were the subject of large international collaborative studies and were demonstrated to quantitatively determine the concentrations of biotech crops in samples of ground grain. Quantitative ELISA methods are also useful for analysis of processed fractions of agricultural commodities such as soybean toasted meal or corn flour. Both strip tests and ELISAs for biotech crops are currently being used on a large scale in the United States to manage the sale and distribution of grain. In these applications, tests are used to determine if the concentration of biotech grain is above or below specified threshold limits. Using existing U.S. Department of Agriculture sampling techniques, the reliability of the threshold determination is expressed in terms of statistical confidence rather than analytical precision. Combining the use of protein immunoassays with Identity Preservation systems provides an effective means of characterizing the raw and processed agricultural inputs to the food production system in a way that allows food producers to comply with labeling laws. PMID: 12083275 [PubMed - indexed for MEDLINE] 1799: J AOAC Int. 2002 May-Jun;85(3):775-9. Production of certified reference materials for the detection of genetically modified organisms. Trapmann S, Schimmel H, Kramer GN, Van den Eede G, Pauwels J. European Commission, Joint Research Centre, Institute for Reference Materials and Measurements, Geel, Belgium. trapmann@irmm.jrc.be Certified reference materials (CRMs) are an essenIial tool in the quality assurance of analytical measurements. They are produced, certified, and used in accordance with relevant ISO (International Organization for Standardization) and BCR (Community Bureau of Reference) guidelines. The Institute for Reference Materials and Measurements (IRMM; Geel, Belgium) has produced the first powdery genetically modified organism (GMO) CRMs in cooperation with the Institute for Health and Consumer Protection (Ispra, Italy). Until now, different weight percentages in the range of 0-5% for 4 GMOs in Europe were produced and certified: Bt (Bacillus thuringiensis)-11 and Bt-176 maize, Roundup Ready soybean, and MON810 maize. Bt-11 and Bt-176 maize and Roundup Ready soybean were produced by IRMM on behalf of Fluka Chemie AG (Buchs, Switzerland). Characterization of used base material is the first step in production and is especially important for GMO CRMs. The production of powdery GMO CRMs and methods used for production control are described. Thorough control of homogeneity and stability are essential for certification of reference materials and ensure validity of the certificate for each bottle of a batch throughout a defined shelf-life. Because production of reference materials and their maintenance are very labor- and cost-intensive tasks, the usefulness of new types of GMO CRMs must be estimated carefully. PMID: 12083274 [PubMed - indexed for MEDLINE] 1800: J AOAC Int. 2002 May-Jun;85(3):768-74. Detection of genetically modified crops and their derivatives: critical steps in sample preparation and extraction. Terry CF, Harris N, Parkes HC. BioAnalytical Innovation Team, LGC, Teddington, Middlesex, UK. catherine.terry@lgc.co.uk The detection of genetically modified crops in foodstuff relies on detection of transgenic DNA or protein material in the sample matrix. Purified DNA or proteins are used as analytical material for polymerase chain reaction technologies and immunodiagnostics. Successful sample preparation is critical to the validity of subsequent analysis. For routine analysis, a good sample preparation technique should be simple, safe, and inexpensive while reproducibly generating DNA/protein of sufficient quality and yield. The suitability of isolated DNA or protein as an analyte for a detection or characterization technique depends on amount or concentration, purity, and integrity, each of which may be influenced by sample matrix and the extraction technique, and, in turn, may impact the validity of analytical techniques. The key sample preparation steps of homogenization, pretreatment, extraction, and purification are discussed as well as typical analytical methods. Consideration is given to application of these steps for particular sample matrixes to maximize yield, reduce inhibition effects, and minimize contamination. The choice of the most appropriate and valid methods for sample preparation from particular foods is discussed with respect to DNA analysis. Attention is also given to ease of use, cost, and generic applicability of the procedures. Publication Types: Research Support, Non-U.S. Gov't PMID: 12083273 [PubMed - indexed for MEDLINE] 1801: J AOAC Int. 2002 May-Jun;85(3):762-7. Identity preservation of genetically modified organisms in the food chain: requirements, methods, and costs. Brookes G. PG Economics, Dorchester, Dorset, UK. Graham.Brookes@Btinternet.com The use of the technology of genetic modification (GM) in European agriculture and the food supply chain is currently controversial. Because of strong anti-GM technology sentiments, the use of ingredients derived from plants containing GM have largely been eliminated from foods manufactured for direct human consumption by the food supply chain in much of the European Union (EU). During the past year, the attention of those opposed to the technology has turned to the use of GM ingredients in livestock production systems by incorporation of GM soy and maize in animal feed. A discussion is presented of the key issues relating to this subject, focusing on how supplies of GM or non-GM products are segregated or how their identities are preserved. The discussion is centered on GM maize and soybeans into which agronomic traits, such as herbicide tolerance and/or insect resistance, have been incorporated. These are currently the only crops into which some varieties containing GM have been approved for use in the EU. PMID: 12083272 [PubMed - indexed for MEDLINE] 1802: J AOAC Int. 2002 May-Jun;85(3):757-61. Analytical challenges: bridging the gap from regulation to enforcement. Van den Eede G, Kay S, Anklam E, Schimmel H. European Commission, Joint Research Centre, Institute for Health and Consumer Protection, Food Products Unit, Ispra, VA, Italy. guy.van-den-eede@jrc.it An overview is presented of the analytical steps that may be needed to determine the presence of genetically modified organisms (GMOs) or for analysis of GMO-derived produce. The analytical aspects necessary for compliance with labeling regulations are discussed along with bottlenecks that may develop when a plant product or a food sample is analyzed for conformity with current European Union GMO legislation. In addition to sampling and testing, other topics deal with complications that arise from biological and agricultural realities that may influence testing capabilities. The issues presented are intended to serve as elements to examine the different challenges that enforcement laboratories might face. PMID: 12083271 [PubMed - indexed for MEDLINE] 1803: J AOAC Int. 2002 May-Jun;85(3):754-6. Method development in relation to regulatory requirements for detection of GMOs in the food chain. Anklam E, Neumann DA. European Commission, Joint Research Centre, Food Products Unit, Ispra, Italy. Elke.anklam@jrc.it This is a summary report of a joint workshop held in Brussels, Belgium, in December 2000. The workshop was organized by the ILSI Europe Novel Food Task Force in collaboration with the European Commission's Joint Research Centre (JRC) and ILSI International Food Biotechnology Committee. The purpose was to investigate progress in the development of analytical methods since the last workshop was held in June 1998. PMID: 12083270 [PubMed - indexed for MEDLINE] 1804: J AOAC Int. 2002 May-Jun;85(3):646-53. Real-time quantitative polymerase chain reaction methods for four genetically modified maize varieties and maize DNA content in food. Brodmann PD, Ilg EC, Berthoud H, Herrmann A. Kantonales Laboratorium Basel-Stadt, Basel, Switzerland. peter.brodmann@kl.bs.ch Quantitative detection methods are needed for enforcement of the recently introduced labeling threshold for genetically modified organisms (GMOs) in food ingredients. This labeling threshold, which is set to 1% in the European Union and Switzerland, must be applied to all approved GMOs. Four different varieties of maize are approved in the European Union: the insect-resistant Bt176 maize (Maximizer), Btl 1 maize, Mon810 (YieldGard) maize, and the herbicide-tolerant T25 (Liberty Link) maize. Because the labeling must be considered individually for each ingredient, a quantitation system for the endogenous maize content is needed in addition to the GMO-specific detection systems. Quantitative real-time polymerase chain reaction detection methods were developed for the 4 approved genetically modified maize varieties and for an endogenous maize (invertase) gene system. PMID: 12083257 [PubMed - indexed for MEDLINE] 1805: Amino Acids. 2002;22(3):259-78. Engineering of cysteine and methionine biosynthesis in potato. Nikiforova V, Kempa S, Zeh M, Maimann S, Kreft O, Casazza AP, Riedel K, Tauberger E, Hoefgen R, Hesse H. Department of Prof. Willmitzer, Max-Planck-Institut für Molekulare Pflanzenphysiologie, Golm, Germany. Methionine and cysteine, two amino acids containing reduced sulfur, are not only an important substrate of protein biosynthesis but are also precursors of various other metabolites such as glutathione, phytochelatines, S-adenosylmethionine, ethylene, polyamines, biotin, and are involved as methyl group donor in numerous cellular processes. While methionine is an essential amino acid due to an inability of monogastric animals and human beings to synthesise this metabolite, animals are still able to convert methionine consumed with their diet into cysteine. Thus, a balanced diet containing both amino acids is necessary to provide a nutritionally favourable food or feed source. Because the concentrations of methionine and cysteine are often low in edible plant sources, e.g. potato, considerable efforts in plant breeding and research have been and are still performed to understand the physiological, biochemical, and molecular mechanisms that contribute to their synthesis, transport, and accumulation in plants. During the last decade molecular tools have enabled the isolation of most of the genes involved in cysteine and methionine biosynthesis, and the efficient plant transformation technology has allowed the creation of transgenic plants that are altered in the activity of individual genes. The physiological analysis of these transgenic plants has contributed considerably to our current understanding of how amino acids are synthesised. We focused our analysis on potato (Solanum tuberosum cv. Désirée) as this plant provides a clear separation of source and sink tissues and, for applied purposes, already constitutes a crop plant. From the data presented here and in previous work we conclude that threonine synthase and not cystathionine gamma-synthase as expected from studies of Arabidopsis constitutes the main regulatory control point of methionine synthesis in potato. This article aims to cover the current knowledge in the area of molecular genetics of sulfur-containing amino acid biosynthesis and will provide new data for methionine biosynthesis in solanaceous plants such as potato. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 12083069 [PubMed - indexed for MEDLINE] 1806: Bull World Health Organ. 2002;80(4):336-8. UN to help developing countries assess safety of GM crops. Hagmann M. Publication Types: News PMID: 12075375 [PubMed - indexed for MEDLINE] 1807: Toxicol Sci. 2002 Jul;68(1):4-8. Approaches to assessment of the allergenic potential of novel proteins in food from genetically modified crops. Kimber I, Dearman RJ. Syngenta Central Toxicology Laboratory, Alderley Park, Macclesfield, Cheshire SK10 4TJ, United Kingdom. ian.kimber@syngenta.com The safety assessment of food derived from genetically modified plants continues to attract considerable attention. Among the important issues that need to be considered is whether the products of novel genes introduced into crop plants will have the potential to induce allergic sensitization or to elicit allergic disease. Hierarchical approaches to allergenicity testing have been proposed, and these incorporate evaluation of the structural and sequence homology and serological identity of novel proteins with known allergens, measurement of resistance to proteolytic digestion, and assessment of allergenic potential using animal models. Accounts of these approaches are available elsewhere, and it is not the purpose of this article to provide a detailed critique of specific methods. Our intention is rather to look more broadly at the strategy for assessment of allergenic potential, the challenges such assessments pose for the practicing toxicologist, and how some of these might best be addressed. Publication Types: Review PMID: 12075104 [PubMed - indexed for MEDLINE] 1808: Tex Med. 2002 Jun;98(6):50-2. Bon appétit. Ortolon K. ken.ortolon@texmed.org PMID: 12073902 [PubMed - indexed for MEDLINE] 1809: Appetite. 2002 Jun;38(3):189-97. Public perception of a range of potential food risks in the United Kingdom. Kirk SF, Greenwood D, Cade JE, Pearman AD. Nutrition Epidemiology Group, Nuffield Institute for Health, 71-75 Clarendon Road, University of Leeds LS2 9PL, UK. s.f.l.kirk@leeds.ac.uk This study aimed to use a standard questionnaire to obtain a nationally representative sample of opinions on a range of potential food risks. Participants were a national sample of 1182 subjects selected using three different approaches: random and sentinel postal samples and a telephone survey. A modified psychometric questionnaire (the Perceived Food Risk Index) was administered to subjects on three occasions, spanning five time-points. Baseline data collection was undertaken from October to December 1998 (phase 1). The second wave of data collection was undertaken over three time-points in February, April and July 1999 (one-third of respondents to phase 1 at each time-point - data combined as phase 2), and the final phase of data collection was between October and December 1999 (phase 3). Principal components analysis was used to assess the intercorrelations between the items on the questionnaire. Two main components were identified as 'dread' and 'knowledge'. Saturated fats were perceived as the least dreaded and the most known of the potential risks considered, while bovine spongiform encephalopathy and Salmonella were the risks dreaded the most. There was a slight perception that the potential risks had become more known over the year, especially for growth hormones. This study has raised a number of important issues for risk communicators. Despite current policy aimed at reducing fat intake, this will be difficult to achieve at a population level since people are not worried about its impact, yet food safety continues to be a significant concern to the public. Copyright 2002 Elsevier Science Ltd. All rights reserved. Publication Types: Research Support, Non-U.S. Gov't PMID: 12071684 [PubMed - indexed for MEDLINE] 1810: J Am Coll Nutr. 2002 Jun;21(3 Suppl):212S-217S. Foods as production and delivery vehicles for human vaccines. Korban SS, Krasnyanski SF, Buetow DE. Department of Natural Resources & Environmental Sciences, University of Illinois, Urbana 61801, USA. s-korban@uiuc.edu Vaccination is a great asset for eradication of infectious diseases in humans and animals. With the prevalence of antibiotic resistant bacterial strains and an alarming increase in new and re-emerging pathogens, the need for vaccination continues to be a high priority for mammalian diseases. In the last several years, a novel approach for developing improved mucosal subunit vaccines has emerged by exploiting the use of genetically modified plants. It has been demonstrated that plant-derived antigens are functionally similar to conventional vaccines and can induce neutralizing antibodies in mammalian hosts. Using genetically engineered plants for the production of immunogenic peptides also provides a new approach for the delivery of a plant-based subunit vaccine, i.e., oral delivery, provided these immunogenic peptides are expressed in an edible part of the plant, such as grain or fruit. Thus, food crops can play a significant new role in promoting human health by serving as vehicles for both production and delivery of vaccines. Publication Types: Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Review PMID: 12071307 [PubMed - indexed for MEDLINE] 1811: J Am Coll Nutr. 2002 Jun;21(3 Suppl):199S-204S. Increased production of nutriments by genetically engineered crops. Sévenier R, van der Meer IM, Bino R, Koops AJ. Business Unit Cell Cybernetics, Plant Research International, Wageningen, The Netherlands. Plants are the basis of human nutrition and have been selected and improved to assure this purpose. Nowadays, new technologies such as genetic engineering and genomics approaches allow further improvement of plants. We describe here three examples for which these techniques have been employed. We introduced the first enzyme involved in fructan synthesis, the sucrose sucrose fructosyltransferase (isolated from Jerusalem artichoke), into sugar beet. The transgenic sugar beet showed a dramatic change in the nature of the accumulated sugar, 90% of the sucrose being converted into fructan. The use of transgenic sugar beet for the production and isolation of fructans will result in a more efficient plant production system of fructans and should promote their use in human food. The second example shows how the over-expression of the key enzyme of flavonoid biosynthesis could increase anti-oxidant levels in tomato. Introduction of a highly expressed chalcone isomerase led to a seventyfold increase of the amount of quercetin glucoside, which is a strong anti-oxidant in tomato. We were also able to modify the essential amino acid content of potato in order to increase its nutritional value. The introduction of a feedback insensitive bacterial gene involved in biosynthesis of aspartate family amino acids led to a sixfold increase of the lysine content. Because the use of a bacterial gene could appear to be controversial, we also introduced a mutated form of the plant key enzyme of lysine biosynthesis (dihydrodipicolinate synthase) in potato. This modification led to a 15 times increase of the lysine content of potato. This increase of the essential amino acid lysine influences the nutritional value of potato, which normally has low levels of several essential amino acids. These three examples show how the metabolism of primary constituents of the plant cell such as sugar or amino acids, but also of secondary metabolites such as flavonoids, can be modified by genetic engineering. Producing fructan, a soluble fiber, increasing the level of flavonoids, an antioxidant, in tomato or increasing the level of essential amino acids in potato are all clear examples of plant genetic modifications with possible positive effects on human nutrition. Publication Types: Review PMID: 12071305 [PubMed - indexed for MEDLINE] 1812: J Am Coll Nutr. 2002 Jun;21(3 Suppl):184S-190S. Fighting iron deficiency anemia with iron-rich rice. Lucca P, Hurrell R, Potrykus I. Institute for Plant Science ETHZ, Zurich, Switzerland. paola.lucca@irb.unisi.ch OBJECTIVE: Iron deficiency is estimated to affect about 30% of the world population. Iron supplementation in the form of tablets and food fortification has not been successful in developing countries, and iron deficiency is still the most important deficiency related to malnutrition. Here we present experiments that aim to increase the iron content in rice endosperm and to improve its absorption in the human intestine by means of genetic engineering. METHODS: We first introduced a ferritin gene from Phaseolus vulgaris into rice grains, increasing their iron content up to twofold. To increase iron bioavailability, we introduced a thermo-tolerant phytase from Aspergillus fumigatus into the rice endosperm. In addition, as cysteine peptides are considered major enhancers of iron absorption, we over-expressed the endogenous cysteine-rich metallothionein-like protein. RESULTS: The content of cysteine residues increased about sevenfold and the phytase level in the grains about one hundred and thirtyfold, giving a phytase activity sufficient to completely degrade phytic acid in a simulated digestion experiment. CONCLUSIONS: This rice, with higher iron content, rich in phytase and cysteine-peptide has a great potential to substantially improve iron nutrition in those populations where iron deficiency is so widely spread. PMID: 12071303 [PubMed - indexed for MEDLINE] 1813: J Am Coll Nutr. 2002 Jun;21(3 Suppl):174S-177S. The impact of consumer food biotechnology training on knowledge and attitude. Santerre CR, Machtmes KL. Foods and Nutrition, 4-H Program, Purdue University, West Lafayette, Indiana 47907-1264, USA. santerre@purdue.edu OBJECTIVE: Consumer education is an important aspect in the adoption of any new technology. The objective of this work was to determine whether consumer's knowledge and attitudes would be influenced by a face-to-face presentation involving food biotechnology. MATERIALS AND METHODS: Participants (576) were requested to complete a pre-test prior to receiving a 45-80 minute presentation, which was then followed by a post-test. Participants included members from a community organization, undergraduate and graduate college students and cooperative extension educators (county agents). RESULTS: Following training, 98% to 99% correctly indicated that fruits and vegetables contain chromosomes and that foods from biotech crops were currently sold in grocery stores. Prior to training, only 31% felt that these crops were properly regulated by federal agencies, and only 25% were confident that bioengineering was unlikely to make an existing food allergenic. Following training, 83% felt that these crops were properly regulated, and 63% believed that biotechnology was unlikely to add new allergens to our food supply. In addition, 90% of those trained would eat or serve genetically-modified foods to their family, and 90% believed that they or their family would benefit from genetically-modified foods within the next five years. CONCLUSIONS: It is apparent from these results that when provided sound, science-based information, participants are more accepting of this technology and the regulatory process. Publication Types: Research Support, Non-U.S. Gov't PMID: 12071301 [PubMed - indexed for MEDLINE] 1814: US News World Rep. 2002 Jun 3;132(19):61. Engineered to run wild. Genetically altered animals could be released to fight pests and disease. Boyce N. Publication Types: News PMID: 12066459 [PubMed - indexed for MEDLINE] 1815: Science. 2002 Jun 14;296(5575):1948-9. Genetically modified food. TV drama sparks scientific backlash. Shouse B. Publication Types: News PMID: 12065809 [PubMed - indexed for MEDLINE] 1816: Trends Biotechnol. 2002 Jul;20(7):291-6. Where will the wood come from? Plantation forests and the role of biotechnology. Fenning TM, Gershenzon J. Max Planck Institute for Chemical Ecology, Winzerlaer Str. 10, D-07745 Jena, Germany. fenning@ice.mpg.de Wood is almost as important to humanity as food, and the natural forests from which most of it is harvested from are of enormous environmental value. However, these slow-growing forests are unable to meet current demand, resulting in the loss and degradation of forest. Plantation forests have the potential to supply the bulk of humanity's wood needs on a long-term basis, and so reduce to acceptable limits the harvest pressures on natural forests. However, if they are to be successful, plantation forests must have a far higher yield of timber than their natural counterparts, on much shorter rotation times. To achieve this in reasonable time, biotechnology must be applied to the tree-improvement process, for which large increases in public and private capital investment are needed. However, additional obstacles exist in the form of opposition to plantations, some forest ecocertification schemes, and concerns about aspects of forest biotechnology, especially genetic engineering. It is the intention of this article to explain, in detail, why plantation forests are needed to sustainably meet the world's demand for wood, why they are not being developed fast enough, and why the application of biotechnology to tree improvement is essential to speeding up this process. PMID: 12062973 [PubMed - indexed for MEDLINE] 1817: Trends Biotechnol. 2002 Jul;20(7):285-6. GM foods in Spanish newspapers. Ramón D, Dorcey E, Gil JV, Serrano A. Publication Types: Letter PMID: 12062971 [PubMed - indexed for MEDLINE] 1818: Proc Biol Sci. 2002 Jun 7;269(1496):1173-6. Sensitive dependencies and separation distances for genetically modified herbicide-tolerant crops. Perry JN. Plant and Invertebrate Ecology Division, IACR Rothamsted Experimental Station, Harpenden, Herts AL5 2JQ, UK. joe.perry@bbsrc.ac.uk The amount of land available for the coexistent growing of both organic and genetically modified herbicide-tolerant (GMHT) crops depends on the separation distance between the two types of crop. The form of the decline in the proportion of land available for growing one of these crop types due to increasing separation distance is linear on a suitable scale, but with a slope and intercept that are sensitively dependent on the proportion of the other crop already present. Spatially explicit simulations from realistic scenarios indicate that a major increase in separation distances, currently under review by the UK government, may have serious implications for the future coexistence of organic and GMHT crops in the UK. Publication Types: Research Support, Non-U.S. Gov't PMID: 12061962 [PubMed - indexed for MEDLINE] 1819: Am J Pathol. 2002 Jun;160(6):2259-65. Life span extension by reduction in growth hormone-insulin-like growth factor-1 axis in a transgenic rat model. Shimokawa I, Higami Y, Utsuyama M, Tuchiya T, Komatsu T, Chiba T, Yamaza H. Department of Respiratory and Digestive Medicine, Division of Experimental Medicine, Pathology, and Gerontology, Nagasaki University School of Medicine, Nagasaki City, Japan. shimo@net.nagasaki-u.ac.jp The longer life span in dwarf mice suggests that a reduction in the growth hormone (GH)-insulin-like growth factor (IGF)-1 axis retards aging and extends the life span in mammals. We tested this hypothesis in a transgenic strain of rats whose GH gene was suppressed by an anti-sense GH transgene. Male rats homozygous for the transgene (tg/tg) had a reduced number of pituitary GH cells, a lower plasma concentration of IGF-1, and a dwarf phenotype. Heterozygous rats (tg/-) had an intermediate phenotype in plasma IGF-1, food intake, and body weight between tg/tg and control (-/-) rats. The life span of tg/tg rats was 5 to 10% shorter than -/- rats. In contrast, the life span of tg/- rats was 7 to 10% longer than -/- rats. Pathological analysis suggested that neoplasms caused earlier death in tg/tg rats; in contrast, tg/- rats had reduced nonneoplastic diseases and a prolonged life span. Immunological analysis revealed a smaller population and lower activity of splenic natural killer cells in tg/tg rats. The results of the present study support the hypothesis, but suggest that there is an optimal level of the GH-IGF-1 axis to maximize survival in mammals. Publication Types: Research Support, Non-U.S. Gov't PMID: 12057928 [PubMed - indexed for MEDLINE] 1820: Tijdschr Diergeneeskd. 2002 May 15;127(10):322-30. [Biotechnology, especially genetic modification, and legislation] [Article in Dutch] de Sitter H, Peters PW. Biotechnology and genetic modification (GM) related legislation is not yet fully developed in the European Union (EU). New legislation has been recently issued ('Introduction of GMO's in the environment') and recently proposals from the European Commission ('GMO's in food and feed' and 'Traceability and labelling of GMO's') entered the decision-making process in the end of 2001. The proposals for the establishment of the European Food Authority play a role in this respect. GMO legislation is complex not in the least because of the demands for the dossiers, to be submitted with an application, while these procedures for admission must become more transparent. In this paper the relevant legislation will be discussed with the exception of that related to human health. Because of dissatisfaction with the present legislation, the European Commission in the past years granted no new approvals for introductions on the market of GMO's and for GM novel foods. New legislation should suspend the present de-facto moratorium. The tasks and position of the Inspectorate for the Health Protection and Veterinary Public Health is discussed. A provision has been made in the legislation with respect to adventitious or technically unavoidable contamination of raw materials with GMO's up to a maximum of 1%, of which the enforcement is not yet watertight. The analytical methods are being still developed. Publication Types: English Abstract PMID: 12056264 [PubMed - indexed for MEDLINE] 1821: Transgenic Res. 2002 Apr;11(2):199-214. Expression of biotin-binding proteins, avidin and streptavidin, in plant tissues using plant vacuolar targeting sequences. Murray C, Sutherland PW, Phung MM, Lester MT, Marshall RK, Christeller JT. The Horticulture and Food Research Institute of New Zealand, Palmerston North Research Centre. Tobacco plants have been developed which constitutively express high levels of the biotin-binding proteins, avidin and streptavidin. These plants were phenotypically normal and produced fertile pollen and seeds. The transgene was expressed and its product located in the vacuoles of most cell types in the plants. Targeting was achieved by use of N-terminal vacuolar targeting sequences derived from potato proteinase inhibitors which are known to target constitutively to vacuoles in potato tubers and, under wound-induction, in tomato leaves. Avidin was located in protein body-like structures within the vacuole and transgene protein levels remained relatively constant throughout the lifetime of the leaf. We describe two chimeric constructs with similar levels of expression. One comprised a potato proteinase inhibitor I signal peptide cDNA sequence attached to an avidin cDNA and the second a potato proteinase inhibitor II signal peptide genomic sequence (including an intron) attached to a core streptavidin synthetic sequence. We were unable to regenerate plants when transformation used constructs lacking the targeting sequences. The highest levels observed (up to 1.5% of total leaf protein) confirm the vacuole as the organelle of choice for stable storage of plant-toxic transgene products. The efficient targeting of these proteins did not result in any measured changes in plant biotin metabolism. Publication Types: Research Support, Non-U.S. Gov't PMID: 12054353 [PubMed - indexed for MEDLINE] 1822: Transgenic Res. 2002 Apr;11(2):185-98. Avidin expressed in transgenic tobacco leaves confers resistance to two noctuid pests, Helicoverpa armigera and Spodoptera litura. Burgess EP, Malone LA, Christeller JT, Lester MT, Murray C, Philip BA, Phung MM, Tregidga EL. The Horticulture and Food Research Institute of New Zealand Limited, Mt Albert Research Center, Auckland. eburgess@hort.cri.nz Fertile transgenic tobacco plants with leaves expressing avidin in the vacuole have been produced and shown to halt growth and cause mortality in larvae of two noctuid lepidopterans, Helicoverpa armigera and Spodoptera litura. Late first instar H. armigera larvae and neonate (< 12-h-old) S. litura larvae placed on leaves excised from T0 tobacco expressing avidin at 3.1-4.6 microM (micromoles/kg of fresh leaf tissue) had very poor growth over their first 8 days on the leaves, significant numbers had died by days 11 or 12 and all were dead by day 22 (H. armigera) or day 25 (S. litura). Similar results were obtained when late first instar H. armigera larvae were placed on leaves from T1 plants expressing avidin at six different average concentrations, ranging from 3.7 to 17.3 microM. Two larvae on the lowest expressing leaves survived to pupation, but there was total mortality among the other groups and no relationship between avidin concentration and the effects on the larvae. Synergistic effects between avidin-expressing tobacco plants and a purified Bt toxin, Cry1Ba, were demonstrated. Late instar H. armigera larvae fed with leaves from T2 plants expressing avidin at average concentrations of either <5.3 or > 12.9 microM, and painted with Cry1Ba protein at a rate equivalent to an expression level of 0.5% of total leaf protein, died significantly faster than larvae given either of the two treatments alone. Larvae fed with avidin-expressing leaves painted with the protease inhibitor, aprotinin, at a rate equivalent to 1% of total leaf protein had mortality similar to those given avidin-leaves alone. There was no evidence of antagonism between these two proteins. Publication Types: Research Support, Non-U.S. Gov't PMID: 12054352 [PubMed - indexed for MEDLINE] 1823: Transgenic Res. 2002 Apr;11(2):161-73. The expression of a mammalian proteinase inhibitor, bovine spleen trypsin inhibitor in tobacco and its effects on Helicoverpa armigera larvae. Christeller JT, Burgess EP, Mett V, Gatehouse HS, Markwick NP, Murray C, Malone LA, Wright MA, Philip BA, Watt D, Gatehouse LN, Lövei GL, Shannon AL, Phung MM, Watson LM, Laing WA. The Horticulture and Food Research Institute of New Zealand, Palmerston North Research Centre. jchristeller@hort.cri.nz The cDNA for bovine spleen trypsin inhibitor (SI), a homologue of bovine pancreatic trypsin inhibitor (BPTI), including the natural mammalian presequence was expressed in tobacco using Agrobacterium tumefaciens-mediated transformation. Stable expression required the N-terminal targeting signal presequence although subcellular localization was not proven. SI was found to exist as two forms, one coinciding with authentic BPTI on western blots and the second marginally larger due to retention of the C-terminal peptide. Both were retained on a trypsin-agarose affinity gel and had inhibitory activity. Newly emergent leaves contained predominantly the large form whereas senescent leaves had little except the fully processed form present. Intermediate-aged leaves showed a gradual change indicating that a slow processing of the inhibitor peptide was occurring. The stability of SI was shown by the presence of protein at high levels in completely senescent leaves. Modifications to the cDNA (3' and 5' changes and minor codon changes) resulted in a 20-fold variation in expression. Expression of modified SI in transgenic tobacco leaves at 0.5% total soluble protein reduced both survival and growth of Helicoverpa armigera larvae feeding on leaves from the late first instar. In larvae surviving for 8 days, midgut trypsin activity was reduced in SI-tobacco fed larvae, while chymotrypsin activity was increased. Activities of leucine aminopeptidase and elastase-like chymotrypsin remained unaltered. The use of SI as an insect resistance factor is discussed. Publication Types: Research Support, Non-U.S. Gov't PMID: 12054350 [PubMed - indexed for MEDLINE] 1824: Toxicol Lett. 2002 Feb 28;127(1-3):351-7. Experience with environmental issues in GM crop production and the likely future scenarios. Gaugitsch H. Federal Environment Agency, Spittelauer Laende 5, A 1090, Vienna, Austria. gaugitsch@ubavie.gv.at In the Cartagena Protocol on Biosafety, standards for risk assessment of genetically modified organisms (GMOs) have been set. The criteria and information basis for the risk assessment of GMOs have been modified by the EU Directive 2001/18/EC. Various approaches to further improve the criteria for environmental risk assessment of GMOs are described in this study. Reports on the ecological impacts of the cultivation of certain non-transgenic crop plants with novel or improved traits as analogy models to transgenic plants showed that the effects of agricultural practice can be at least equally important as the effects of gene transfer and invasiveness, although the latter currently play a major role in risk assessment of transgenic crops. Based on these results the applicability of the methodology of 'Life Cycle Analysis (LCA)' for genetically modified plants in comparison with conventionally bred and organically grown crop plants was evaluated. The methodology was regarded as applicable with some necessary future improvements. In current projects, the assessment of toxicology and allergenicity of GM crops are analysed, and suggestions for standardization are developed. Based on results and recommendations from these efforts there are still the challenges of how to operationalize the precautionary principle and how to take into account ecologically sensitive ecosystems, including centres of origin and centres of genetic diversity. Publication Types: Review PMID: 12052677 [PubMed - indexed for MEDLINE] 1825: Toxicol Lett. 2002 Feb 28;127(1-3):341-9. Regulatory control of genetically modified (GM) foods: likely developments. Schilter B, Constable A. Food Safety Group, Nestlé Research Center, Vers-chez-les-Blanc, CH-1000 Lausanne 26, Switzerland. benoit.schilter@rdls.nestle.com The placing of genetically modified (GM) crops on the European market requires a regulatory approval supported by a thorough safety evaluation. This approach has been applied to all GM crops presently on the market. Despite this stringent process there has been an increasing public concern about the impact of GM foods on human health and the environment. In this context, regulatory control may develop in several directions. One response to the public concern is to strengthen the data requirements for the risk assessment process. Several avenues have been proposed. They include the application of technologies such as proteomics and metabolomics to assess unintended changes, and the development of predictive methods to evaluate allergenicity. Obligations for post-launch surveillance have appeared in regulations. Criteria are required to define when and why such approaches are necessary. Significant challenges including feasibility and validation of the methods, and safety relevance of the data generated will have to be addressed before any general application of these new approaches. Effective monitoring requires the ability to identify the presence of GM products and trace their origin. Traceability and labeling are therefore important developments in the GM food regulatory arena. Both require the development of reliable analytical detection tools. PMID: 12052676 [PubMed - indexed for MEDLINE] 1826: Toxicol Lett. 2002 Feb 28;127(1-3):337-40. Clinical risk assessment of GM foods. Lack G. Department of Paediatric Allergy and Immunology, Imperial College at St Mary's Hospital, Praed Street, London W2 1NY, UK. gideon.lack@st-marys.nhs.uk The main concerns about adverse effects of genetically modified (GM) foods on health are the transfer of antibiotic resistance, toxicity and allergenicity. There are two issues from an allergic standpoint. First, the transfer of a known allergen may occur from a crop into a non-allergenic target crop. The second scenario is the creation of a neo-allergen where de novo sensitisation occurs in the population. The first scenario occurred in 1996 when the 2S albumen protein from Brazil nut was transferred into soy bean (N. Engl. J. Med. 334 (1996) 688). 2S albumen was found to be a major Brazil nut allergen and the newly expressed protein in transgenic soy retained its allergenicity. Patients allergic to Brazil nuts and not to soy bean now showed an IgE mediated response towards GM soy bean. We argue that it is possible to prevent such occurrences by doing IgE-binding studies and taking into account physico-chemical characteristics of proteins and referring to known allergen databases. The second possible scenario of de novo sensitisation does not easily lend itself to risk assessment. We compare GM technology to traditional plant breeding and food processing methods. There is no evidence that the technology used for the production of GM foods poses an allergic threat per se compared to other methodologies widely accepted in the food industry. We need to proceed cautiously in the future, assessing individual GM foods on the basis of their individual merits and risks prior to introducing them into the market. PMID: 12052675 [PubMed - indexed for MEDLINE] 1827: Toxicol Lett. 2002 Feb 28;127(1-3):329-36. Current and future benefits from the use of GM technology in food production. Engel KH, Frenzel T, Miller A. Technische Universität München, Lehrstuhl für Allgemeine Lebensmitteltechnologie, Am Forum 2, D-85350 Freising-Weihenstephan, Germany. k.h.engel@lrz.tu-muenchen.de For the current generation of genetically modified (GM) crops the improvement of agronomic traits (e.g. herbicide tolerance, insect resistance) has been a major objective. The lack of obvious and direct benefits for the consumer has been a main point of criticism. Future trends will increasingly encompass the modification of quality traits, such as the improvement of sensory and especially nutritional properties. Some of the ongoing developments try to meet the desire of consumers for 'healthy' or 'high-tech' foods in developed countries. Others are intended to assist in adjusting the nutritional status of foods to the needs of consumers in developing countries. Considering the increasing world population and the limited amount of arable land, GM technology may also become a valuable tool to ensure food security. The major prerequisite for the applicability of the technique is the safety of the resulting products. The increasing complexity of modifications intended might require adjustments and improvements of the strategies applied to the safety assessment of GM foods. Present research activities try to meet these new challenges. PMID: 12052674 [PubMed - indexed for MEDLINE] 1828: Toxicol Lett. 2002 Feb 28;127(1-3):307-13. Ethical issues for bioscientists in the new millennium. Purchase IF. Institute of Medicine, Law and Bioethics, School of Biological Sciences, The University of Manchester, Oxford Road, M13 9PT, UK. ifhp@chadzombe.u-net.com The scientific understanding of biological processes is developing extremely fast, providing opportunities for changing people's lives in many ways-through health care, food and the environment. The speed with which these changes are occurring means that even bioscientists can only keep up with their own narrow field of science. It is not surprising that members of the public are frightened about the rapidity and impact of the changes arising from the biological revolution. These concerns are often expressed in ethical terms. Decision making about the direction of research and its application is becoming more transparent. This means that bioscientists will have to engage in the debate about their work with members of the public, including those who are opposed to it, in order to create acceptance of their work and its products. At the moment, bioscientists are often ill equipped to enter this debate because of their lack of training in ethics and lack of understanding of the impact of ethics on their work. A better understanding of bioethics will be necessary for entering this debate with vigour. A comprehensive ethical analysis is outside the scope of this text. Some of the principal arguments about the ethics of two aspects of bioscience research-genetically modified crops and the use of experimental animals-will be discussed to illustrate a few of the issues that derive from ethical analyses. I hope that this will encourage toxicologists to take a greater interest in bioethics. Publication Types: Review PMID: 12052671 [PubMed - indexed for MEDLINE] 1829: Mutat Res. 2002 Jun;511(2):89-112. Modulation of heterocyclic amine-induced mutagenicity and carcinogenicity: an 'A-to-Z' guide to chemopreventive agents, promoters, and transgenic models. Dashwood RH. Department of Environmental and Molecular Toxicology, Linus Pauling Institute, and Department of Environmental and Molecular Toxicology, Oregon State University, Corvallis 97331-6512, USA. rod.dashwood@orst.edu A landmark report by Widmark in 1939 describing "cancer-producing substances in roasted food", and the seminal work of Sugimura and colleagues in the 1970s on the isolation of potent mutagens from cooked meat and fish stimulated a major international effort on the study of heterocyclic amines and their modulators. The latter term is used in its broadest context to mean agents or conditions that positively or negatively influence the mutagenic or carcinogenic activities of heterocyclic amines in vitro or in vivo. An 'A-to-Z' list of these modulators includes well over 150 natural or synthetic phytochemicals, micronutrients and antioxidants, as well as several large chemical classes (polyphenols, flavones, retinoids, porphyrins), food fractions, and food preparation methods. In many cases, the findings reported in the literature can be regarded as descriptive, but for a number of specific agents there is sufficient evidence to glean some understanding of the inhibitory or promotional mechanisms of action. These mechanisms can be divided into 11 separate sub-categories, arranged within a general classification scheme that encompasses such terms as 'blocking agents', 'suppressing agents', 'desmutagens', 'bioantimutagens', 'interceptor molecules' and 'tumor promoters'. In addition, new research directions, most notably during the past 2-3 years or so, have led to the use of novel dosing protocols and unique animal models (including transgenic species) that provide insight into exposure conditions and genetic background as modulators of heterocyclic amine activity in vitro and in vivo. Overall, the more than 250 citations on the subject give ample evidence of the growing interest in modulators of heterocyclic amine carcinogenesis and mutagenesis, and their possible importance in determining human cancer risk in defined populations. Publication Types: Research Support, U.S. Gov't, P.H.S. Review PMID: 12052429 [PubMed - indexed for MEDLINE] 1830: Cas Lek Cesk. 2002 Mar 1;141(4):107-11. [Genetically modified organisms--problems and legislation] [Article in Czech] Drobník J. drobnik@mbox.cesnet.cz Genetically modified organisms are defined by law as entities capable of replication and/or transmission of hereditary material that had been altered by the insertion or removal of a DNA fragment. By the EU legal regulation as well as by the Czech law, such organisms are considered risky whereas other products of breeding, though obtained by, e.g., induced mutagenesis, are claimed as safe. Organisms transferred from other ecosystems are also considered safe. The Czech law on the use of genetically modified organisms is based on registers of users and organisms for specific use. Application for the registration that is valid as an approval should be submitted to the Ministry of Environment. The applicant is obliged to present the risk assessment of the particular use of genetically modified organisms. Genetically modified organisms are connected with certain risk to ecology, however health risks are brought about almost exclusively by microorganisms. Modified organisms used for food production are thoroughly tested for substantial equivalency with standard crops and with respect to health parameters of the protein(s) newly introduced due to genetic modification. Detail tests as well as their cost are close to the testing of new drugs. European as well as Czech rules for food labelling are motivated by the psychology of consumers rather than by health impact. They result to absurdities but do not meet the task of public psychology. This is why the EU authorities are looking for measures to change the present situation that other wise would bring Europe well behind the developed countries. Publication Types: English Abstract PMID: 12046253 [PubMed - indexed for MEDLINE] 1831: Curr Opin Allergy Clin Immunol. 2002 Jun;2(3):249-52. Genetically engineered foods: implications for food allergy. Taylor SL, Hefle SL. University of Nebraska, Food Allergy Research and Resource Program, Lincoln, Nebraska 68583-0919, USA. staylor2@unl.edu The products of agricultural biotechnology, including such common foods as corn and soybeans, are already reaching the consumer marketplace. Consumer exposure to such foods is already fairly significant, particularly in the USA. Thus far, no reports exist regarding allergic reactions to the crops that have been approved for introduction into the food supply. These crops have been modified to only a minor extent by comparison with their traditional counterparts, and the level of expression of new and novel proteins is quite low. Thus, consumer exposure to these novel proteins is very low and unlikely to result in allergic sensitization. Nevertheless, foods produced through agricultural biotechnology must be assessed for safety, including their potential allergenicity, before they may be approved by worldwide regulatory agencies for entry into the food supply. However, the adequacy of the current approach to the assessment of the potential allergenicity of foods produced through agricultural biotechnology has been the subject of considerable scientific and regulatory debate. Publication Types: Review PMID: 12045422 [PubMed - indexed for MEDLINE] 1832: J Clin Invest. 2002 Jun;109(11):1429-36. Hypothalamic growth hormone secretagogue receptor regulates growth hormone secretion, feeding, and adiposity. Shuto Y, Shibasaki T, Otagiri A, Kuriyama H, Ohata H, Tamura H, Kamegai J, Sugihara H, Oikawa S, Wakabayashi I. Department of Medicine, Nippon Medical School, Tokyo, Japan. shuto@nms.ac.jp Growth hormone secretagogues (GHSs) stimulate GH secretion and food intake. GHS receptor (GHS-R) mRNA has been identified mainly in the arcuate nucleus (Arc) and ventromedial nucleus of the hypothalamus and in the pituitary. Ghrelin, an endogenous ligand for GHS-R, has recently been purified from rat stomach. Although ghrelin is also expressed in the hypothalamus, the physiological significance of the ghrelin/GHS-R system is still unknown. We have created transgenic (Tg) rats expressing an antisense GHS-R mRNA under the control of the promoter for tyrosine hydroxylase (TH), thus selectively attenuating GHS-R protein expression in the Arc. Tg rats had lower body weight and less adipose tissue than did control rats. Daily food intake was reduced, and the stimulatory effect of GHS treatment on feeding was abolished in Tg rats. GH secretion and plasma insulin-like growth factor-I levels were reduced in female Tg rats. These results suggest that GHS-R in the Arc is involved in the regulation of GH secretion, food intake, and adiposity. Publication Types: Research Support, Non-U.S. Gov't PMID: 12045256 [PubMed - indexed for MEDLINE] 1833: Nat Biotechnol. 2002 Jun;20(6):537-41. Comment in: Nat Biotechnol. 2002 Aug;20(8):775-6. Liabilities and economics of transgenic crops. Smyth S, Khachatourians GG, Phillips PW. University of Saskatchewan, Saskatoon, SK, Canada. sjs064@mail.usask.ca PMID: 12042844 [PubMed - indexed for MEDLINE] 1834: Nat Biotechnol. 2002 Jun;20(6):527. Comment in: Nat Biotechnol. 2002 Sep;20(9):871. Going with the flow. [No authors listed] Publication Types: Editorial PMID: 12042836 [PubMed - indexed for MEDLINE] 1835: J Nutr. 2002 Jun;132(6):1384-90. Food biotechnology: benefits and concerns. Falk MC, Chassy BM, Harlander SK, Hoban TJ 4th, McGloughlin MN, Akhlaghi AR. Life Sciences Research Office, 9650 Rockville Pike, Bethesda, MD 20814, USA. falkm@isro.faseb.org Recent advances in agricultural biotechnology have highlighted the need for experimental evidence and sound scientific judgment to assess the benefits and risks to society. Nutrition scientists and other animal biologists need a balanced understanding of the issues to participate in this assessment. To date most modifications to crop plants have benefited producers. Crops have been engineered to decrease pesticide and herbicide usage, protect against stressors, enhance yields and extend shelf life. Beyond the environmental benefits of decreased pesticide and herbicide application, consumers stand to benefit by development of food crops with increased nutritional value, medicinal properties, enhanced taste and esthetic appeal. There remains concern that these benefits come with a cost to the environment or increased risk to the consumer. Most U.S. consumers are not aware of the extent that genetically modified foods have entered the marketplace. Consumer awareness of biotechnology seems to have increased over the last decade, yet most consumers remain confused over the science. Concern over the impact on the safety of the food supply remains low in the United States, but is substantially elevated in Europe. Before a genetically engineered crop is introduced into commerce it must pass regulatory scrutiny by as many as four different federal regulatory bodies to ensure a safe food supply and minimize the risk to the environment. Key areas for more research are evaluation of the nutritional benefits of new crops, further investigation of the environmental impact, and development of better techniques to identify and track genetically engineered products. Publication Types: Research Support, Non-U.S. Gov't PMID: 12042463 [PubMed - indexed for MEDLINE] 1836: Nutr Rev. 2002 May;60(5 Pt 1):135-41. Genetically engineered crops: their potential use for improvement of human nutrition. Yan L, Kerr PS. Department of Nutritional Sciences, Du Pont Protein Technologies, St. Louis, Missouri 63188, USA. Great success has been achieved in increasing agriculture productivity to fulfill human needs during the second half of the 20th century. However, there will be much greater challenges in the future. Based on the current population growth rate of 1.4% per year, the world's population is forecast to increase from the current level of approximately six billion to nine to twelve billion in 50 years. In addition to continuously increasing demand for agricultural production, there is an urgent need to improve the nutritional quality of human diets for this rapidly growing human population. Malnutrition is still a worldwide health issue. Macronutrient and micronutrient deficiencies are prevalent in developing countries, and over-consumption of certain nutrients in developed countries (e.g., saturated fatty acids) is associated with high incidence of certain chronic diseases. Furthermore, there will be declining natural resources such as arable land and water, and the challenges to humans must be met without further degrading the environment. Biotechnology offers a valuable tool to help achieve these goals. This review focuses on the most recent advances in biotechnology, which promise to improve human nutrition by enhancing the nutrient density of plant foods. Issues relating to the safety of food products from genetically engineered crops are also discussed. Publication Types: Review PMID: 12030276 [PubMed - indexed for MEDLINE] 1837: Ann N Y Acad Sci. 2002 May;964:185-96. Prediction of allergenicity of gene-modified foods by serum-based testing. Poulsen LK. Laboratory of Medical Allergology, National University Hospital, DK-2100 Copenhagen, Denmark. lkpallgy@inet.uni2.dk On the basis of applying the IFBC/ILSI decision tree in a number of cases, a refinement of the scheme is suggested. Large differences in allergenic potential may be obtained by altering the route of administration of an allergen. Because an inhalation allergen can induce symptoms at different threshold doses depending on whether it is introduced via the inhalation or oral route, we propose that double-blind, placebo-controlled food challenges be performed in all cases of inhalation allergens being present in foods. Even proteins never previously ingested may cause IgE binding and elicit clinically relevant symptoms. Publication Types: Review PMID: 12023205 [PubMed - indexed for MEDLINE] 1838: Ann N Y Acad Sci. 2002 May;964:173-83. The dog as a model for food allergy. Buchanan BB, Frick OL. Department of Plant and Microbial Biology, University of California, Berkeley 94720, USA. view@nature.berkeley.edu Research during the past decade has shown the dog to be an excellent model for human food allergies. Humans and dogs share many of the same allergies to foods. Furthermore, the dog model shows clinical symptoms typical of humans, that is, both experience vomiting and diarrhea. Present results suggest that the dog may provide a means to test genetically modified foods for unsuspected allergens. Publication Types: Review PMID: 12023204 [PubMed - indexed for MEDLINE] 1839: Ann N Y Acad Sci. 2002 May;964:151-61. Assessment of protein allergenicity: studies in brown norway rats. Knippels LM, Penninks AH. Experimental Immunology, Department of Target Organ Toxicology, TNO Nutrition and Food Research, 3700 AJ Zeist, The Netherlands. knippels@voeding.tno.nl For the safety evaluation of genetically engineered crops, the potential allergenicity of the newly introduced protein(s) has become an important issue. There is, however, no universal and reliable test system for the evaluation of the allergic sensitizing ability of food proteins. Therefore, there is a growing interest in the development of animal models. This paper summarizes the results of a promising food allergy model developed in Brown Norway (BN) rats. The results demonstrate that BN rats can be sensitized via the relevant oral route of exposure. Daily gavage dosing of the animals with several food proteins, without the use of adjuvants, resulted in significant antigen-specific IgE responses. In addition, the profile of allergens recognized by the immune system of the BN rat, appeared comparable to the profile of allergens recognized by allergic humans. Besides oral sensitization, local and systemic immune-mediated effects, such as increased gastrointestinal permeability, decreased breathing frequency, and decreased blood pressure, could also be observed in the sensitized animals after an oral challenge. All together, these observations suggest that this BN rat model might provide a suitable animal model to study the allergenicity of food proteins in humans. Publication Types: Review PMID: 12023202 [PubMed - indexed for MEDLINE] 1840: Ann N Y Acad Sci. 2002 May;964:129-38. Phosphomannose isomerase, a novel plant selection system: potential allergenicity assessment. Privalle LS. Syngenta Seeds, Inc., Research Triangle Park, North Carolina 27705, USA. Laura.privalle@syngenta.com Phosphomannose isomerase (PMI), an enzyme not present in many plants, catalyzes the reversible interconversion of mannose 6-phosphate and fructose 6-phosphate. Plant cells lacking this enzyme are incapable of surviving on synthetic medium containing mannose. Thus PMI/mannose selection has utility in the identification of transformed plant cells. As part of the safety assessment transgenic plants undergo before commercialization, PMI has been evaluated for its potential allergenicity. Purified PMI protein was readily digestible in a simulated gastric environment. PMI has no sequence homology to known allergens, does not contain multiple disulfide bonds, and has no N-glycosylation consensus sequences. No detectable changes in glycoprotein profiles were detected in PMI-transformed plants as compared to nontransgenic controls. These results indicate that PMI lacks many of the attributes associated with known oral allergens. PMID: 12023200 [PubMed - indexed for MEDLINE] 1841: Curr Biol. 2002 May 14;12(10):R337. Kernel knowledge. Harris RF. Publication Types: News PMID: 12015127 [PubMed - indexed for MEDLINE] 1842: Planta. 2002 May;215(1):79-89. Epub 2002 Jan 23. 4-Hydroxycinnamoyl-CoA hydratase/lyase, an enzyme of phenylpropanoid cleavage from Pseudomonas, causes formation of C(6)-C(1) acid and alcohol glucose conjugates when expressed in hairy roots of Datura stramonium L. Mitra A, Mayer MJ, Mellon FA, Michael AJ, Narbad A, Parr AJ, Waldron KW, Walton NJ. Food Safety Science Division, Institute of Food Research, Norwich Resesrch Park, Colney, Norwich NR4 7UA, UK. 4-Hydroxycinnamoyl-CoA hydratase/lyase (HCHL), a crotonase homologue of phenylpropanoid catabolism from Pseudomonas fluorescens strain AN103, led to the formation of 4-hydroxybenzaldehyde metabolites when expressed in hairy root cultures of Datura stramonium L. established by transformation with Agrobacterium rhizogenes. The principal new compounds observed were the glucoside and glucose ester of 4-hydroxybenzoic acid, together with 4-hydroxybenzyl alcohol- O-beta- D-glucoside. In lines actively expressing HCHL, these together amounted to around 0.5% of tissue fresh mass. No protocatechuic derivatives were found, although a trace of vanillic acid-beta- D-glucoside was detected. There was no accumulation of 4-hydroxybenzaldehydes, whether free or in the form of their glucose conjugates. There was some evidence suggesting a diminished availability of feruloyl-CoA for the production of feruloyl putrescine and coniferyl alcohol. The findings are discussed in the context of a diversion of phenylpropanoid metabolism, and the ability of plants and plant cultures to conjugate phenolic compounds. Publication Types: Research Support, Non-U.S. Gov't PMID: 12012244 [PubMed - indexed for MEDLINE] 1843: Plant Physiol. 2002 May;129(1):122-33. Overexpression of polygalacturonase in transgenic apple trees leads to a range of novel phenotypes involving changes in cell adhesion. Atkinson RG, Schröder R, Hallett IC, Cohen D, MacRae EA. The Horticulture and Food Research Institute of New Zealand, Mount Albert Research Centre, Private Bag 92 169, Auckland, New Zealand. Polygalacturonases (PGs) cleave runs of unesterified GalUA that form homogalacturonan regions along the backbone of pectin. Homogalacturonan-rich pectin is commonly found in the middle lamella region of the wall where two adjacent cells abut and its integrity is important for cell adhesion. Transgenic apple (Malus domestica Borkh. cv Royal Gala) trees were produced that contained additional copies of a fruit-specific apple PG gene under a constitutive promoter. In contrast to previous studies in transgenic tobacco (Nicotiana tabacum) where PG overexpression had no effect on the plant (K.W. Osteryoung, K. Toenjes, B. Hall, V. Winkler, A.B. Bennett [1990] Plant Cell 2: 1239-1248), PG overexpression in transgenic apple led to a range of novel phenotypes. These phenotypes included silvery colored leaves and premature leaf shedding due to reduced cell adhesion in leaf abscission zones. Mature leaves had malformed and malfunctioning stomata that perturbed water relations and contributed to a brittle leaf phenotype. Chemical and ultrastructural analyses were used to relate the phenotypic changes to pectin changes in the leaf cell walls. The modification of apple trees by a single PG gene has offered a new and unexpected perspective on the role of pectin and cell wall adhesion in leaf morphology and stomatal development. Publication Types: Research Support, Non-U.S. Gov't PMID: 12011344 [PubMed - indexed for MEDLINE] 1844: Plant Physiol. 2002 May;129(1):95-102. Direct interference with rhamnogalacturonan I biosynthesis in Golgi vesicles. Skjøt M, Pauly M, Bush MS, Borkhardt B, McCann MC, Ulvskov P. Biotechnology Group, Danish Institute of Agricultural Sciences, Thorvaldsensvej 40, 1871 Copenhagen, Denmark. Pectin is a class of complex cell wall polysaccharides with multiple roles during cell development. Assigning specific functions to particular polysaccharides is in its infancy, in part, because of the limited number of mutants and transformants available with modified pectic polymers in their walls. Pectins are also important polymers with diverse applications in the food and pharmaceutical industries, which would benefit from technology for producing pectins with specific functional properties. In this report, we describe the generation of potato (Solanum tuberosum L. cv Posmo) tuber transformants producing pectic rhamnogalacturonan I (RGI) with a low level of arabinosylation. This was achieved by the expression of a Golgi membrane-anchored endo-alpha-1,5-arabinanase. Sugar composition analysis of RGI isolated from transformed and wild-type tubers showed that the arabinose content was decreased by approximately 70% in transformed cell walls compared with wild type. The modification of the RGI was confirmed by immunolabeling with an antibody recognizing alpha-1,5-arabinan. This is the first time, to our knowledge, that the biosynthesis of a plant cell wall polysaccharide has been manipulated through the action of a glycosyl hydrolase targeted to the Golgi compartment. Publication Types: Comparative Study Research Support, Non-U.S. Gov't PMID: 12011341 [PubMed - indexed for MEDLINE] 1845: J Agric Food Chem. 2002 May 22;50(11):3161-4. Detection of genetically modified coho salmon using polymerase chain reaction (PCR) amplification. Masri S, Rast H, Ripley T, James D, Green M, Jia X, Devlin RH. CFIA, Centre for Plant Health, 8801 East Saanich Road, Sidney, British Columbia V8L 1H3, Canada. masris@inspection.gc.ca A PCR-based protocol for the identification of genetically modified salmon carrying a growth hormone transgene was developed. Several primer pairs were examined, and the primers that gave consistent results were selected to conduct routine testing. Comparison among several DNA extraction procedures, as well as different buffer compositions, led to the adoption of TriZol as the method of choice. Low potassium and high magnesium chloride concentrations were very important in the overall success of the PCR reaction, whereas buffer pH, ranging from 8.3 to 9.2, had little impact on the amplification reaction. The optimal primer annealing temperature was 52 degrees C. Although fish muscle tissues were the primary source for DNA samples, detection of the transgene was also possible in bones, skin, fins, and other organs. No benefits were achieved by the addition of additives such as dimethyl sulfoxide and betaine to the PCR reaction. This optimized PCR method was used to identify all samples tested (61 samples and 17 controls) with 100% accuracy. PMID: 12009980 [PubMed - indexed for MEDLINE] 1846: Proc Nutr Soc. 2002 Feb;61(1):31-7. Mad cows, mad corn and mad communities: the role of socio-cultural factors in the perceived risk of genetically-modified food. Finucane ML. Decision Research, Eugene, Oregon 97401, USA. melissafinucane@hotmail.com The rapid globalization of the world economy has increased the need for a knowledge base of reliable socio-cultural differences in perceptions, values and ways of thinking about new food technologies. Awareness of socio-cultural differences is important because collaborative efforts to deal with food hazards presuppose some understanding of where, how and why the viewpoints of various stakeholders may differ. In the present paper factors that influence public perceptions of genetically-modified (GM) food are discussed, with a special focus on the unique circumstances of populations in the USA, Europe and developing countries. It is argued that effective communication and decision making about the risk of GM food depends critically on understanding how socio-cultural groups differ in their values and in the way they deal with the risks and benefits of new technologies. The implications of psychological aspects of perceived risk (including the roles of qualitative dimensions of risk, world views and trust) for public acceptance of new food technologies are highlighted. Publication Types: Review PMID: 12002793 [PubMed - indexed for MEDLINE] 1847: Proc Nutr Soc. 2002 Feb;61(1):25-9. Genetically modified foods, science, consumers and the media. Rowland IR. Northern Ireland Centre for Diet and Health, School of Biomedical Sciences, University of Ulster, Coleraine, UK. I.Rowlands@ulst.ac.uk In contrast to the situation in the USA, where a wide range of genetically modified (GM) foods is available, in Europe very few GM products have been approved for marketing as foods, and there is widespread public concern about their safety and environmental impact. The marketing of a GM crop for food use in Europe falls under the EC novel foods regulations, and applications require the submission of an extensive dossier of information. The safety evaluation of GM foods presents considerable problems both in the conduct and interpretation of experimental studies, because conventional toxicity tests used in the evaluation of simple chemicals may not be appropriate for whole foods. To rationalise the safety evaluation process and to circumvent the difficulties in toxicological assessment of food materials, the concept of substantial equivalence has been developed. The concept is that if it can be demonstrated that the novel food is essentially similar to its conventional counterpart in terms of critical nutritional or anutritional components, then it is likely to be no more or less toxic than the latter. The possible introduction of unintended effects by the genetic modification process is particularly problematic for the safety evaluation process. The new genomic and post-genomic techniques are potentially valuable in the safety evaluation of GM foods, although they are as yet in their infancy. Publication Types: Review PMID: 12002791 [PubMed - indexed for MEDLINE] 1848: Pediatr Allergy Immunol. 2002 Apr;13(2):73-4. Genetically modified food and the pediatric allergist. Warner JO. Publication Types: Editorial PMID: 12000476 [PubMed - indexed for MEDLINE] 1849: Shokuhin Eiseigaku Zasshi. 2002 Feb;43(1):24-9. [A detection method of recombinant DNA from genetically modified potato (NewLeaf Plus potato) and detection of NewLeaf Plus potato in snack] [Article in Japanese] Akiyama H, Sugimoto K, Matsumoto M, Isuzugawa K, Shibuya M, Goda Y, Toyoda M. National Institute of Health Sciences: 1-18-1, Kamiyoga, Setagaya-ku, Tokyo 158-8501, Japan. A detection method using polymerase chain reaction (PCR) was developed to detect the genetically modified (GM) potato (NewLeaf Plus potato; NL-P), which has not been authorized as safe in foods in Japan. The potato sucrose synthase gene was used as an internal control. The DNA from NL-P specifically provided an amplified band using PCR with a primer pair recognizing PLRV-rep gene. In addition, to prevent false-positive results in processed potato foods infected with PLRV, we designed a primer pair recognizing sequences derived from two organisms to detect specifically NL-P in processed potato. The PCR product obtained using the designed primer pair was specific for NL-P. The DNA introduced into NL-P could be detected from potato powder samples containing 0.05% NL-P. The proposed method was applied to the detection of NL-P in 25 processed potato foods. NL-P was detected in 3 snack products. Publication Types: English Abstract Research Support, Non-U.S. Gov't PMID: 11998315 [PubMed - indexed for MEDLINE] 1850: Cell Biol Toxicol. 2002;18(1):1-11. Toxicity of argemone oil: effect on hsp70 expression and tissue damage in transgenic Drosophila melanogaster (hsp70-lacZ) Bg9. Mukhopadhyay I, Nazir A, Mahmood K, Saxena DK, Das M, Khanna SK, Chowdhuri DK. Embryotoxicology Section, Industrial Toxicology Research Centre, Lucknow, India. The effect of argemone oil on hsp70 expression and tissue damage was investigated by studying beta-galactosidase activity, Western blotting and hybridization, and trypan blue staining in the larval tissues of transgenic Drosophila melanogaster (hsp70-lacZ)Bg9. Different concentrations of argemone oil were mixed with food and third-instar larvae were allowed to feed on them for different time intervals (2, 4, 24, and 48 h). Argemone oil was found to induce hsp70 even in the lowest concentration of the adulterant while maximum tissue damage was observed in the higher two treatment groups. Malpighian tubules and midgut tissue reflected maximum damage as evidenced by both high beta-galactosidase activity and trypan blue staining in these tissues. A prior temperature shock treatment to the larvae was enough to protect the larvae from argemone oil-induced tissue damage as evidenced by little or no trypan blue staining. The present study suggests the cytotoxic potential of argemone oil and further strengthens the evidence for the use of hsp70 as a biomarker in risk assessment. Publication Types: Research Support, Non-U.S. Gov't PMID: 11991082 [PubMed - indexed for MEDLINE] 1851: J Environ Sci Health B. 2002 Jan;37(1):93-101. Nutritional assessment of transgenic sweetpotato on body weight, lipid, and protein status in hamsters. Shireen KF, Pace RD. Department of Food and Nutritional Sciences, Tuskegee University, AL 36088, USA. Kanizs@hotmail.com The objectives of the present study were to evaluate the nutritional quality of genetically modified sweetpotato (genotype PI318846-3) on growth, lipid metabolism, and protein metabolism of hamsters. Three different diets made with transgenic and nontransgenic sweetpotato protein flour including a control diet with casein were fed to male Golden Syrian hamsters for 28 days. The protein efficiency ratio (1.35 +/- 0.01) of the transgenic sweetpotato protein diet was significantly higher (p<0.05) than the nontransgenic sweetpotato and control diets. Plasma albumin and plasma total protein concentrations of hamsters fed the sweetpotato diets were significantly lower (p<0.05) than that of the control. The casein diet (control) produced hypercholesterolemia in hamsters, whereas sweetpotato diets maintained lower plasma and liver total and LDL-cholesterol concentrations in hamsters. Sweetpotatoes contain less amount of protein to maintain the normal animal growth; however, transgenic sweetpotato has good quality protein that supported the growth of hamsters better than nontransgenic sweetpotato. PMID: 11990363 [PubMed - indexed for MEDLINE] 1852: Tech Vasc Interv Radiol. 2001 Jun;4(2):99-106. Alteplase and tenecteplase: applications in the peripheral circulation. Semba CP, Sugimoto K, Razavi MK; Society of Cardiovascular and Interventional Radiology (SCVIR). Cardiovascular Clinical Research, Genentech Inc., MS 59, 1 DNA Way, South San Francisco, CA 94080-4990, USA. Alteplase (t-PA), a recombinant analogue of human tissue plasminogen activator, became the first genetically engineered thrombolytic approved by the Food and Drug Administration in 1987 for acute myocardial infarction (AMI). In addition to AMI, alteplase is currently approved for the treatment of acute ischemic stroke and pulmonary embolism, and we anticipate approval for catheter clearance in late 2001 in a 2-mg vial configuration. With the withdrawal of human neonatal kidney cell-derived urokinase, alteplase has become an alternative agent in peripheral vascular applications. Because few interventionalists had prior experience with the handling and dosage of alteplase, the Advisory Panel to the Society of Cardiovascular and Interventional Radiology established practice guidelines for use in noncoronary applications. Emerging clinical experience with contemporary dosing regimens shows a safety and efficacy profile similar to urokinase but with significantly reduced drug costs. Tenecteplase (TNK) is a genetically modified version of alteplase. TNK is the only plasminogen activator available that has shown a significantly enhanced safety profile versus alteplase in AMI. Approved for a 5-second, single-bolus injection in AMI, TNK possesses a longer half-life, increased resistance to plasminogen activator inhibitor, and improved fibrin specificity compared with alteplase. Because of its enhanced safety profile, TNK may be a desirable agent for peripheral vascular applications. Initial clinical studies with TNK in acute arterial and venous disease are ongoing. This article outlines the Advisory Panel guidelines for using alteplase and highlights features of tenecteplase. Copyright 2001 by W.B. Saunders Company Publication Types: Guideline Practice Guideline Review PMID: 11981795 [PubMed - indexed for MEDLINE] 1853: QJM. 2002 May;95(5):335-6. Technophobia. Berry C. PMID: 11978907 [PubMed - indexed for MEDLINE] 1854: Curr Opin Allergy Clin Immunol. 2001 Jun;1(3):269-71. Hidden food allergens. Hefle SL. Food Allergy Research and Resource Program, University of Nebraska, Lincoln, Nebraska, USA. shefle1@unl.edu This review summarizes recent advances and findings in the area of 'hidden' food allergens, i.e. allergenic foods that can either contaminate other foods, or be 'disguised' as part of a food, and cause allergic reactions. Newly emerging allergenic foods of increasing importance, recently developed methods for the detection of allergenic residues, the potential allergenicity of genetically engineered foods, and some unexpected sources of food allergens are described. Publication Types: Review PMID: 11964700 [PubMed - indexed for MEDLINE] 1855: Expert Rev Mol Diagn. 2002 Jan;2(1):69-76. Traceability of genetically modified organisms. Aarts HJ, van Rie JP, Kok EJ. h.j.m.aarts@rikilt.wag-ur.nl EU regulations stipulate the labeling of food products containing genetically modified organisms (GMOs) unless the GMO content is due to adventitious and unintended 'contamination' and not exceeding the 1% level at ingredient basis. In addition, member states have to ensure full traceability at all stages of the placing on the market of GMOs. Both requirements ensure consumers 'right to know', facilitate enforcement of regulatory requirements and are of importance for environmental monitoring and postmarket surveillance. Besides administrative procedures, such as used in quality certification systems, the significance of adequate molecular methods becomes more and more apparent. During the last decade a considerable number of molecular methods have been developed and validated that enable the detection, identification and quantification of GMO impurities. Most of them rely on the PCR technology and can only detect one specific stretch of DNA. It can, however, be anticipated that in the near future the situation will become more complex. The number of GMO varieties, including 'stacked-gene' varieties, which will enter the European Market will increase and it is likely that these varieties will harbor more variable constructs. New tools will be necessary to keep up with these developments. One of the most promising techniques is microarray analysis. This technique enables the screening for a large number of different GMOs within a single experiment. Publication Types: Review PMID: 11963810 [PubMed - indexed for MEDLINE] 1856: Mol Biol Cell. 2002 Apr;13(4):1329-37. Regulated disruption of inositol 1,4,5-trisphosphate signaling in Caenorhabditis elegans reveals new functions in feeding and embryogenesis. Walker DS, Gower NJ, Ly S, Bradley GL, Baylis HA. Department of Zoology, University of Cambridge, Cambridge CB2 3EJ, United Kingdom. Inositol 1,4,5-trisphosphate (IP(3)) is an important second messenger in animal cells and is central to a wide range of cellular responses. The major intracellular activity of IP(3) is to regulate release of Ca(2+) from intracellular stores through IP(3) receptors (IP(3)Rs). We describe a system for the transient disruption of IP(3) signaling in the model organism Caenorhabditis elegans. The IP(3) binding domain of the C. elegans IP(3)R, ITR-1, was expressed from heat shock-induced promoters in live animals. This results in a dominant-negative effect caused by the overexpressed IP(3) binding domain acting as an IP(3) "sponge." Disruption of IP(3) signaling resulted in disrupted defecation, a phenotype predicted by previous genetic studies. This approach also identified two new IP(3)-mediated processes. First, the up-regulation of pharyngeal pumping in response to food is dependent on IP(3) signaling. RNA-mediated interference studies and analysis of itr-1 mutants show that this process is also IP(3)R dependent. Second, the tissue-specific expression of the dominant-negative construct enabled us to circumvent the sterility associated with loss of IP(3) signaling through the IP(3)R and thus determine that IP(3)-mediated signaling is required for multiple steps in embryogenesis, including cytokinesis and gastrulation. Publication Types: Research Support, Non-U.S. Gov't PMID: 11950942 [PubMed - indexed for MEDLINE] 1857: Curr Opin Biotechnol. 2002 Apr;13(2):142-5. Improving iron, zinc and vitamin A nutrition through plant biotechnology. Zimmermann MB, Hurrell RF. Laboratory of Human Nutrition, Institute of Food Science and Nutrition, Swiss Federal Institute of Technology, Zürich, PO Box 474, CH-8803 Rüschlikon, Switzerland. michael.zimmermann@ilw.agrl.ethz.ch Recent understanding of plant metabolism has made it possible to increase the iron, zinc and beta-carotene (provitamin A) content in staple foods by both conventional plant breeding and genetic engineering. Improving the micronutrient composition of plant foods may become a sustainable strategy to combat deficiencies in human populations, replacing or complementing other strategies such as food fortification or nutrient supplementation. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 11950566 [PubMed - indexed for MEDLINE] 1858: Nature. 2002 Apr 11;416(6881):576-8. Agricultural biotech: the rice squad. Surridge C. Publication Types: News PMID: 11948321 [PubMed - indexed for MEDLINE] 1859: Nature. 2002 Apr 11;416(6881):567. Rice must be perfectly cooked. [No authors listed] Publication Types: Editorial PMID: 11948309 [PubMed - indexed for MEDLINE] 1860: Trends Biotechnol. 2002 May;20(5):215-23. Detection of genetically modified organisms in foods. Ahmed FE. Dept of Radiation Oncology, Leo W. Jenkins Cancer Center, The Brody School of Medicine, LSB 014, East Carolina University, Greenville, NC 27858, USA. ahmedf@mail.ecu.edu Legislation enacted worldwide to regulate the presence of genetically modified organisms (GMOs) in crops, foods and ingredients, necessitated the development of reliable and sensitive methods for GMO detection. In this article, protein- and DNA-based methods employing western blots, enzyme-linked immunosorbant assay, lateral flow strips, Southern blots, qualitative-, quantitative-, real-time- and limiting dilution-PCR methods, are discussed. Where information on modified gene sequences is not available, new approaches, such as near-infrared spectrometry, might tackle the problem of detection of non-approved genetically modified (GM) foods. The efficiency of screening, identification and confirmation strategies should be examined with respect to false-positive rates, disappearance of marker genes, increased use of specific regulator sequences and the increasing number of GM foods. Publication Types: Comparative Study Review PMID: 11943377 [PubMed - indexed for MEDLINE] 1861: Anal Bioanal Chem. 2002 Jan;372(1):230-1. Epub 2001 Dec 11. The role of modern biochemical methods in the determination of hygienically relevant microorganisms in foodstuffs and (drinking) water. Weyandt RG. Publication Types: News PMID: 11939201 [PubMed - indexed for MEDLINE] 1862: BMC Biotechnol. 2002 Apr 4;2:4. Genetically modified parthenocarpic eggplants: improved fruit productivity under both greenhouse and open field cultivation. Acciarri N, Restaino F, Vitelli G, Perrone D, Zottini M, Pandolfini T, Spena A, Rotino G. Research Institute for Vegetable Crops, Monsampolo del Tronto, AP, Italy. acciarri@libero.it BACKGROUND: Parthenocarpy, or fruit development in the absence of fertilization, has been genetically engineered in eggplant and in other horticultural species by using the DefH9-iaaM gene. The iaaM gene codes for tryptophan monoxygenase and confers auxin synthesis, while the DefH9 controlling regions drive expression of the gene specifically in the ovules and placenta. A previous greenhouse trial for winter production of genetically engineered (GM) parthenocarpic eggplants demonstrated a significant increase (an average of 33% increase) in fruit production concomitant with a reduction in cultivation costs. RESULTS: GM parthenocarpic eggplants have been evaluated in three field trials. Two greenhouse spring trials have shown that these plants outyielded the corresponding untransformed genotypes, while a summer trial has shown that improved fruit productivity in GM eggplants can also be achieved in open field cultivation. Since the fruits were always seedless, the quality of GM eggplant fruits was improved as well. RT-PCR analysis demonstrated that the DefH9-iaaM gene is expressed during late stages of fruit development. CONCLUSIONS: The DefH9-iaaM parthenocarpic gene is a biotechnological tool that enhances the agronomic value of all eggplant genotypes tested. The main advantages of DefH9-iaaM eggplants are: i) improved fruit productivity (at least 30-35%) under both greenhouse and open field cultivation; ii) production of good quality (marketable) fruits during different types of cultivation; iii) seedless fruit with improved quality. Such advantages have been achieved without the use of either male or female sterility genes. Publication Types: Research Support, Non-U.S. Gov't PMID: 11934354 [PubMed - indexed for MEDLINE] 1863: FDA Consum. 2001 Jan-Feb;35(1):14-20. A new kind of fish story. The coming of biotech animals. Lewis C. PMID: 11930918 [PubMed - indexed for MEDLINE] 1864: FDA Consum. 2000 Jul-Aug;34(4):7. Plans for more rigorous review of bioengineered foods. [No authors listed] Publication Types: News PMID: 11924640 [PubMed - indexed for MEDLINE] 1865: Nat Biotechnol. 2002 Apr;20(4):324-5. European Union in disarray over GM seeds. Meldolesi A. Publication Types: News PMID: 11923823 [PubMed - indexed for MEDLINE] 1866: Kokuritsu Iyakuhin Shokuhin Eisei Kenkyusho Hokoku. 2001;(119):27-39. [Hypersensitivity about environmental chemicals--mainly about food allergy] [Article in Japanese] Teshima R. rteshima@nihs.go.jp The hypersensitivity of environmental chemicals and natural products has been reviewed. Among environmental chemicals, small molecular weight molecules work as hapten and cause immediate-type and delayed-type hypersensitivity. Among natural products, relatively lower molecular weight protein or glycoprotein (MW 10,000-70,000 kDa) work as allergen and cause mainly immediate-type hypersensitivity. In recent years, amino acid sequence of important natural allergens have been determined, and three-dimensional structure and IgE epitopes of some of these allergens have also been determined. The characteristics of both inhalation and food allergens have been summarized. As for food allergens, the stability of these proteins in simulated gastric fluid(SGF) was one of the most important characteristics. In the last parts, the approach to the assessment of allergenic potential of genetically modified foods has been summarized. Publication Types: English Abstract Review PMID: 11915282 [PubMed - indexed for MEDLINE] 1867: Nature. 2002 Mar 21;416(6878):252. Diplomats near pact in simmering debate over transgenic foods. Giles J. PMID: 11907542 [PubMed - indexed for MEDLINE] 1868: Chembiochem. 2001 Apr 2;2(4):289-90. Scientists and society--and never the twain shall meet? Wright K. Publication Types: Consensus Development Conference Review PMID: 11905466 [PubMed - indexed for MEDLINE] 1869: Tijdschr Diergeneeskd. 2002 Mar 1;127(5):157-60. [Can gene technology in agriculture prevent hunger in the world?] [Article in Dutch] Goewie EA. Departement Plant- en gewaswetenschappen, Universiteit Wageningen, Haarweg 333, 6709 RZ Wageningen. The world population grows rapidly: the number of mouths to feed increases. Is an agriculture without gene technology able to produce sufficiently in order to prevent hunger? Research indicates that hunger is not the result of short comings in agricultural outputs. It is however the result of poverty. This problem will not be solved by gene technology based agricultural production. This article explains the basic principles of mainstream and organic farming. Literature shows that the production potentials of both kinds of farming are, by far most, not yet exhausted. Gene technology is therefore unnecessary. Publication Types: English Abstract PMID: 11905236 [PubMed - indexed for MEDLINE] 1870: J Agric Food Chem. 2002 Mar 27;50(7):2100-9. Detection of recombinant DNA segments introduced to genetically modified maize (Zea mays). Matsuoka T, Kuribara H, Takubo K, Akiyama H, Miura H, Goda Y, Kusakabe Y, Isshiki K, Toyoda M, Hino A. National Food Research Institute, 2-1-12 Kannondai, Tsukuba, Ibaraki 305-8642, Japan. Polymerase Chain Reaction (PCR) techniques are increasingly used for the detection of genetically modified (GM) crops in foods. In this paper, recombinant DNAs introduced into the seven lines of GM maize, such as Event 176, Bt11, T25, MON810, GA21, DLL25, and MON802, are sequenced. On the basis of the obtained sequence, 14 primer pairs for the detection of the segments, such as promoter, terminator regions, and construct genes, were designed. To confirm the specificities of the designed primer pairs, PCR was performed on genomic DNAs extracted from GM and non-GM maize, GM and non-GM soy, and other cereal crops. Because the presence of the corresponding DNA segments was specifically detected in GM crops by the designed primer pairs, it was concluded that this method is useful for fast and easy screening of GM crops including unauthorized ones. Publication Types: Research Support, Non-U.S. Gov't PMID: 11902963 [PubMed - indexed for MEDLINE] 1871: Curr Allergy Asthma Rep. 2002 Jan;2(1):55-62. Biotechnology and food allergy. Helm RM. University of Arkansas for Medical Sciences, Arkansas Children's Hospital Research Institute, 1120 Marshall Street, Little Rock, AR 72202-3591, USA. HelmRickiM@uams.edu The production of genetically modified foods for an increasingly informed and selective consumer requires the coordinated activities of both the companies developing the transgenic food and regulatory authorities to ensure that these foods are at least as safe as the traditional foods they are supplementing in the diet. Although the size and complexity of the food sector ensures that no single player can control the process from seed production through farming and processing to final products marketed in a retail outlet, checks and balances are in place to ensure that transgenic foods will provide a convenient, wholesome, tasty, safe, affordable food source. Ultimately, it is the responsibility of companies developing the genetically modified food to provide relevant data to regulatory agencies, such as the US Department of Agriculture, Environmental Protection Agency, and Food and Drug Administration, to confirm that the transgenic product is reasonably safe for the consumer, as zero risk from allergen sensitization is nonexistent. Publication Types: Review PMID: 11895627 [PubMed - indexed for MEDLINE] 1872: Nature. 2002 Mar 14;416(6877):123. In risk assessment, one has to admit ignorance. Hoffmann-Riem H, Wynne B. Publication Types: Letter PMID: 11894070 [PubMed - indexed for MEDLINE] 1873: Toxicol Pathol. 2002 Jan-Feb;30(1):129-31. Safety assessment and public concern for genetically modified food products: the European view. Moseley BE. bevmos@bmoseley.fsnet.co.uk The safety assessment for marketing purposes of genetically modified (GM) foods in the 15 Member States of the European Union (EU) is based on the Novel Foods and Novel Food Ingredients Regulation adopted in May 1997. Before a GM food can be approved under the Regulation, it must satisfy three criteria: Gm food must be safe, it must not mislead the consumer and it must be nutritionally adequate. The EU Scientific Committee on Food has published a set of guidelines describing the type of information expected from a company in support of an application for approval of a GM food or food ingredient. Despite this rigorous procedure and there being no evidence of harm resulting from the consumption of GM foods worldwide, there is essentially no market in the EU for such products at present. Possible reasons for this are discussed and the view put forward that the market for GM foods will change only when there are more clearly perceived consumer benefits. Publication Types: Review PMID: 11892726 [PubMed - indexed for MEDLINE] 1874: Toxicol Pathol. 2002 Jan-Feb;30(1):132-4. Safety assessments and public concern for genetically modified food products: the American view. Harlander SK. BlOrational Consultants, Inc, New Brighton, Minnesota 55112, USA. SKHARLAND@aol.com In the relatively short time since their commercial introduction in 1996, genetically modified (GM) crops have been rapidly adopted in the United States GM crops are regulated through a coordinated framework developed in 1992 and administered by three agencies-the US Department of Agriculture (USDA) that ensures the products are safe to grow, the Environmental Protection Agency (EPA) that ensures the products are safe for the environment, and the Food and Drug Administration (FDA) that ensures the products are safe to eat. Rigorous food and environmental safety assessments must be completed before GM crops can be commercialized. Fifty-one products have been reviewed by the FDA, including several varieties of corn, soybeans, canola, cotton, rice, sugar beets, potatoes, tomatoes, squash, papaya, and flax. Because FDA considers these crops "substantially equivalent" to their conventional counterparts, no special labeling is required for GM crops in the United States and they are managed as commodities with no segregation or identity preservation. GM crops have thus made their way through commodity distribution channels into thousands of ingredients used in processed foods. It has been estimated that 70% to 85% of processed foods on supermarket shelves in the United States today contain one or more ingredients potentially derived from GM crops. The food industry and retail industry have been monitoring the opinions of their consumers on the GM issue for the past several years. Numerous independent groups have also surveyed consumer concerns about GM foods. The results of these surveys are shared and discussed here. Publication Types: Review PMID: 11890465 [PubMed - indexed for MEDLINE] 1875: Toxicol Pathol. 2002 Jan-Feb;30(1):126-8. Safety assessment and public concerns for genetically modified food products: the Japanese experience. Hino A. Gustatory Biology Laboratory, National Food Research Institute, Tsukuba, Ibaraki, Japan. akhino@nfri.affrc.go.jp The recombinant DNA (rDNA) technique is expected to bring about great progress in the improvement of breeding technology and the development of new plant varieties showing high quality and high yield, such as those with excellent pest and disease resistance, those with environmental stress tolerance, and so forth. In the United States and Canada, many genetically modified (GM) crop plants were commercialized as early as 1994. In Japan, 35 transgenic crop plants, such as herbicide tolerant soybean, cotton, and canola, and insect-resistant corn, cotton, and potatos, were authorized and considered marketable until April 2001. The general public, however, is not familiar with rDNA technology, and some people seem to feel uncomfortable with biotechnology, frequently because of the difficulty of the technology and lacking of sufficient information. New labeling systems were initiated in April 2001 in Japan to provide information regarding the use of GM crops as raw material. Publication Types: Review PMID: 11890464 [PubMed - indexed for MEDLINE] 1876: Toxicol Pathol. 2002 Jan-Feb;30(1):117-25. Current methods for assessing safety of genetically modified crops as exemplified by data on Roundup Ready soybeans. Nair RS, Fuchs RL, Schuette SA. Monsanto Company, St Louis, Missouri 63167, USA. rashmi.s.nair@Monsanto.com Several laboratories have used recombinant DNA technology in plant breeding to improve compositional, processing, and agronomic characteristics of plants. These transformed plants have been extensively tested in field trials, have gained full regulatory approvals and are currently being marketed in a number of countries around the world. This paper briefly summarizes the approach used to assure the safety of foods and feeds derived from these genetically modified crops, as exemplified by data on Roundup Ready soybeans that has been developed by Monsanto Company using biotechnology in order to confer tolerance to glyphosate, the active ingredient in Roundup herbicide, by the production of the CP4 enolpyruvylshikimate-3-phosphate synthase protein. The results of the studies demonstrate that Roundup Ready soybeans are as safe as traditional soybeans with respect to food and feed safety. Publication Types: Review PMID: 11890463 [PubMed - indexed for MEDLINE] 1877: Int J Vitam Nutr Res. 2002 Jan;72(1):7-12. Biotechnology: a solution for improving nutrient bioavailability. King JC. United States Department of Agriculture, Agricultural Research Service, Western Human Nutrition Research Center, University of California, Davis, CA 95616, USA. jking@whnrc.usda.gov Biotechnology strategies are now available to improve the amount and availability of nutrients in plant crops. Those strategies include simple plant selection for varieties with high nutrient density in the seeds, cross-breeding for incorporating a desired trait within a plant, and genetic engineering to manipulate the nutrient content of the plant. In plant cross-breeding, all genes of the parent plants are combined and the progeny have both desirable and undesirable traits. To eliminate undesirable traits, plant breeders "back-cross" the new plant varieties with other plants over several generations. This technique, called hybridization, has been used to create varieties of low-phytate corn, barley, and rice. Using the techniques of genetic engineering, the gene(s) encoding for a desired trait(s) in a plant are introduced in a precise and controlled manner within a relatively short period of time. Golden rice, containing carotenoids, and rice with higher amounts of iron, are two examples of genetically engineered plants for improved nutrition. Genetic engineering has tremendous potential for revolutionizing nutrition. However, public concerns regarding safety, appearance, and ethics must be overcome before these products can be effectively introduced into the food supply. Publication Types: Review PMID: 11887755 [PubMed - indexed for MEDLINE] 1878: Environ Health Perspect. 2002 Mar;110(3):A130-3. Allergies à la carte: is there a problem with genetically modified foods? Eubanks M. Publication Types: News PMID: 11882488 [PubMed - indexed for MEDLINE] 1879: J Agric Food Chem. 2002 Mar 13;50(6):1553-61. Glycoalkaloid content and chemical composition of potatoes improved with nonconventional breeding approaches. Esposito F, Fogliano V, Cardi T, Carputo D, Filippone E. Department of Food Science, University of Naples Federico II, Parco Gussone, 80055 Portici, Italy. This paper reports the results of chemical analyses performed on two distinct groups of new potato genotypes. The first group contained five clones transformed with the gene ech42 encoding for an endochitinase. The second included 21 interspecific hybrids between the cultivated potato Solanum tuberosum and the wild species S. commersonii, obtained either by somatic fusion or by sexual hybridization. Tubers from transgenic plants were analyzed for several morphological and biochemical parameters to ascertain the substantial equivalence between the transgenic genotypes and the original cultivar Désirée. The interspecific hybrids were analyzed for the same parameters in order to identify genotypes with novel improved chemical characteristics and with low levels of glycoalkaloids deriving from the wild species and potentially hazardous to human health. For transgenic tubers, the results provided evidence that indicates the substantial equivalence between the transgenic genotypes and the cultivated control for the considered traits. The results suggest that chitinase gene insertion did not alter other metabolic pathways of potato tubers and did not cause unintentional pleiotropic effects. As far as interspecific hybrids are concerned, wide variability for all of the parameters analyzed was found. For some useful traits (e.g., soluble solids and proteins, dry matter content) the interspecific hybrids performed better than both the cultivated control and the wild species. In a number of genotypes, glycoalkaloid levels were close to or lower than those of the control varieties, suggesting that selection for low glycoalkaloid content is possible. The results also indicated that glycoalkaloids from S. commersonii may be lost rapidly. Indeed, some hybrids were found to have the same glycoalkaloid profile as S. tuberosum. Finally, the results showed that among the parameters considered, glycoalkaloid content is the most sensitive to variation. Therefore, glycoalkaloid determination should be used for routine control of genotypes produced by interspecific hybridization. Publication Types: Research Support, Non-U.S. Gov't PMID: 11879036 [PubMed - indexed for MEDLINE] 1880: Rocz Panstw Zakl Hig. 2001;52(4):313-20. [Usefulness of an immunoassay test TRAIT for detection of genetically modified Roundup ready soybean in food products] [Article in Polish] Urbanek-Karłowska B, Fonberg-Broczek M, Sawilska-Rautenstrauch D, Badowski P, Jedra M. Zakład Badania Zywności i Przedmiotów Uzytku Państwowy Zakład Higieny 00-791 Warszawa. u.l. Chocimska 24 The test based on immunoassay TRAIT Test for the specific detection of Roundup Ready Soybean was used for reference material in the form of dried powdered soy beans contained 0, 0.3, 1.25, 2.5% of genetically modified material, for soy beans declared as Roundup Ready and for soy products from Warsaw market. The detection limit was approximately 0.1% GMO on dry weight basis. Experiment was also carried out on heated soybeans. The positive results was obtained since temperature was under 65 degrees C during 15 minutes of heating grounded beans; above this temperature specific protein was not recognisable by the antibody. The TRAIT Test should be regarded as a qualitative method and could be recommended for screening purposes. Investigation demonstrated that above mentioned test was useful for detection of protein of genetically modified soybean in unprocessed products. Publication Types: English Abstract PMID: 11878012 [PubMed - indexed for MEDLINE] 1881: Shokuhin Eiseigaku Zasshi. 2001 Dec;42(6):367-73. [Comparison of carotenoid components between GM and non-GM papaya] [Article in Japanese] Mutsuga M, Ohta H, Toyoda M, Goda Y. National Institute of Health Sciences: 1-18-1, Kamiyoga, Setagaya-ku, Tokyo 158-8501, Japan. We compared the carotenoid profile in GM papaya (Sunup) line to that of a non-GM one (Sunset). First, to identify major carotenoids in papaya, large-scale extraction was carried out with methanol. HPLC analyses of the methanol extracts revealed that both papayas mainly contained 5 pigments and no apparent difference was observed in the HPLC profiles. On the basis of LC/MS data and photodiode-array spectra, beta-carotene (3), lycopene (2), beta-cryptoxanthin (1), and beta-cryptoxanthin myristoyl and lauroyl esters (4 and 5) were identified as major carotenoids. It is well known that most carotenoids are labile, so a rapid analysis with precautions to avoid decomposition was developed to quantify their contents in the original fruits. Frozen samples of the fruits were sliced and a piece (about 2 g) of fruit was cut out and lyophilized. After extraction of the piece with methanol containing an anti-oxidant, BHT, the extract was further partitioned with hexane and methanol. Finally the contents of the main carotenoids in the hexane fraction were analyzed by HPLC. The contents of total carotenoids (sum of 1-5) and beta-cryptoxanthin (1, 4 and 5) in GM papaya fruit were estimated to be 0.764 +/- 0.056 and 0.604 +/- 0.051 (mumol/g), respectively and those in non-GM fruit were 0.883 +/- 0.145 and 0.705 +/- 0.098 (mumol/g), respectively. These differences are not statistically significant. Publication Types: Comparative Study English Abstract PMID: 11875821 [PubMed - indexed for MEDLINE] 1882: Nature. 2002 Feb 28;415(6875):948-9. Alleged flaws in gene-transfer paper spark row over genetically modified maize. Butler D. Publication Types: News PMID: 11875532 [PubMed - indexed for MEDLINE] 1883: Science. 2002 Mar 1;295(5560):1617-9. Has GM corn 'invaded' Mexico? Mann CC. Publication Types: News PMID: 11872805 [PubMed - indexed for MEDLINE] 1884: Camb Q Healthc Ethics. 2002 Winter;11(1):97-101. Chris Shaw on ethical issues in biotechnology. Interview by Thomasine Kushner. Shaw C. Pharmaceutical Biotechnology Research Group, University of Ulster, Northern Ireland. Publication Types: Interview PMID: 11868425 [PubMed - indexed for MEDLINE] 1885: Eur Rev Med Pharmacol Sci. 2001 Jan-Feb;5(1):25-9. Genetically modified foods and children potential health risks. Cantani A, Micera M. Pediatric Department, University of Roma La Sapienza. AIM: Professor Pusztai was publicly humiliated over claims that genetically modified (GM) Frankenstein food may be harmful. He was stripped of his post and described as 'muddled' by his superiors after he referred to experiments in which rats had been damaged when fed genetically-altered potatoes. Who is in an unsound scenario, supported by verbal expressions ("substantially"), should even more expend further effort in conducting scientific investigation into the safety of GM varieties of plants. OBSERVATIONS: Of particular concern is the exposure of infants and children to GM foods (GMFs) because of their possible increased susceptibility for untoward effects. Several examples stress that the ascertainment of human disease emerged after certain materials were widely used. Studies show that some compounds were not adequately tested for toxicity before their commercial introduction, whereas proper premarked testing would have prevented a prolonged exposure. CONCLUSIONS: Too often the toxicity of these substances is untested and the potential hazards that they may pose to children have not been examined. Nobody has evaluated whether intrauterine and infant exposure to GMFs may have profound permanent and irreversible consequences even in adult life. In this paper we analyse issues pertaining to children's health that have been largely ignored. Publication Types: Review PMID: 11860219 [PubMed - indexed for MEDLINE] 1886: Planta. 2002 Jan;214(3):356-64. High-level production of the non-cariogenic sucrose isomer palatinose in transgenic tobacco plants strongly impairs development. Börnke F, Hajirezaei M, Heineke D, Melzer M, Herbers K, Sonnewald U. Institut für Pflanzengenetik und Kulturpflanzenforschung (IPK), Gatersleben, Germany. boernke@ipk-gatersleben.de Palatinose (isomaltulose, 6-O-alpha-D-glucopyranosyl-D-fructose) is a structural isomer of sucrose which is produced from sucrose by some bacterial strains as a reserve material during periods of low carbon availability. The ability to synthesise palatinose is not only advantageous for the bacteria but is also of industrial interest since palatinose is used as a sucrose substitute in food production. To explore the possibility of palatinose production in plants a recently isolated sucrose isomerase gene (palI; EC 5.4.99.11) from Erwinia rhapontici [F. Börnke et al. (2001) J Bacteriol 183: 2425-2430] was cloned into a plant expression vector between the constitutive 35S CaMV promoter and the octopine synthase polyadenylation signal. To allow secretion of the protein into the apoplast the signal peptide of the potato proteinase inhibitor II was N-terminally fused to the pall coding region. Expression of the protein was verified by northern and western blot analyses. Efficient secretion of the protein was demonstrated by palI detection in intercellular fluids. Transgenic plants expressing palI accumulated high levels of palatinose. As a consequence, transgenic plants showed severe phenotypic alterations. Young leaves were curled and developed bleached areas during maturation. Flowers were misshapen and sterile. Based on nonaqueous fractionation experiments palatinose was found in several subcellular compartments, indicating limited membrane transport of the sugar. In contrast to results obtained with short-term feeding experiments, no evidence for palatinose-mediated regulation of photosynthetic or defence genes could be obtained in the transgenic palI-expressing tobacco plants. Based on our results we conclude that plants can efficiently be used as bioreactors for the production of palatinose. Furthermore, tissue-specific expression of palI should allow carbon allocation to specific tissues and/or cell-types to be modulated. Publication Types: Comparative Study PMID: 11855640 [PubMed - indexed for MEDLINE] 1887: J Agric Food Chem. 2002 Feb 27;50(5):1016-21. Detection of genetically modified maize by the polymerase chain reaction and capillary gel electrophoresis with UV detection and laser-induced fluorescence. García-Cañas V, González R, Cifuentes A. Institute of Industrial Fermentations (CSIC), Juan de la Cierva 3, 28006 Madrid, Spain. In this paper, the possibilities of capillary gel electrophoresis (CGE) to detect transgenic maize in flours are shown. The method is based on the extraction and amplification by the polymerase chain reaction (PCR) of a specific DNA fragment from transgenic maize and its subsequent analysis by CGE with UV detection or laser-induced fluorescence (LIF). Some useful considerations regarding the optimization of DNA extraction and amplification conditions are given. Also, a comparison is established between the two CGE protocols for DNA detection based on ultraviolet absorption (CGE-UV) and LIF (CGE-LIF). The requirements, advantages, and limitations of both CGE methods are discussed. To our knowledge, this is the first paper on the use of CGE-LIF to detect transgenic food. Publication Types: Research Support, Non-U.S. Gov't PMID: 11853473 [PubMed - indexed for MEDLINE] 1888: NCSL Legisbrief. 2001 Aug-Sep;9(33):1-2. International trade of new agricultural products from biotechnology. Naftzger D, Smith S. NCSL--Washington, D.C., USA. david.naftzger@ncsl.org PMID: 11850893 [PubMed - indexed for MEDLINE] 1889: J Food Prot. 2002 Feb;65(2):426-31. Analysis of flour and food samples for cry9C from bioengineered corn. Orlandi PA, Lampel KA, South PK, Assar SK, Carter L, Levy DD. Center for Food Safety and Applied Nutrition, Food and Drug Administration, Washington, DC 20204, USA. StarLink corn is a variety of yellow corn that has been genetically modified by the insertion of an altered cry9C gene into the plant genome. resulting in expression of the insecticidal Cry9C protein. The U.S. Environmental Protection Agency has approved StarLink corn for use in animal feed but not in food intended for human consumption. Therefore, under the U.S. Food, Drug, and Cosmetic Act, any food intended for human consumption in which the presence of StarLink corn is indicated by the presence of either the Cry9C protein or the cry9C gene would be considered adulterated. Extraction and PCR-based methods were used to detect the presence of the cry9C DNA initially in corn flour and corn meal, and then these methods were extended to the analysis of processed corn products, including taco shells, cereals, baby foods, party snacks, and chips, for the presence of this modified genetic material. In a survey of 63 products, the cry9C transgene was detected in 4 taco shells. PMID: 11848580 [PubMed - indexed for MEDLINE] 1890: Cell Mol Biol (Noisy-le-grand). 2001 Dec;47(8):1343-51. The future of transgenic plants in developing countries. Weil A. Cirad, Paris, France. alain.weil@cirad.fr Whatever their own policies may be, developing countries will inevitably be affected by the development of genetically-modified organisms in industrialized countries. While maintaining a cautious attitude, most of these countries wish to keep their options open, thus protecting themselves from the risk of being deprived of future technologies that might allow them to achieve self-sufficiency in food production, to resolve certain problems confronting their most vulnerable populations and to preserve the international competitiveness of their products. Companies should see that it is in their interest to help these countries implement their own policies, notably through an open attitude to industrial property. If the value of genetic engineering is thus confirmed, then it perhaps in this manner that GMOs will earn the legitimacy required to make them acceptable to the people of Northern countries where the majority of solvent markets are located. Publication Types: Review PMID: 11838954 [PubMed - indexed for MEDLINE] 1891: Cell Mol Biol (Noisy-le-grand). 2001 Dec;47(8):1329-42. Identity, traceability, acceptability and substantial equivalence of food. Pascal G, Mahé S. INRA, Direction Scientifique Nutrition Humaine et Sécurité des Aliments (DS-NHISA), Paris, France. hermet@paris.inra.fr The numerous food crises that Europe has experienced during the past five years have raised new consumer demands concerning the characterization, traceability, and safety of foods which are proposed on the market. The consumer has, at the same time, vigorously placed into question the modes of agricultural production in industrialized countries, as well as the structures and means of evaluating the food risks and the conditions of the consumer's participation in the public debate in these domains. For certain groups of consumers, one also attends a contestation of the expertise and the application to the food domain of the considerable progress that has taken place in the field of biotechnology. So it is that the development of genetically modified organisms (mainly plants, the raw material of food products) has experienced a slowing down in the European Union. The answers afforded to these new exigencies of consumers in matter of identity, traceability, and acceptability of the foods are dealt with in this paper, as well as the elements which may concur with the evaluation of their safety. The positive role that biotechnology can afford to the different domains is emphasized. A source of uneasiness, biotechnology is also a powerful tool for ameliorating the evaluation of the sanitary risks and for answering the hopes of the citizen in the food domain. Publication Types: Review PMID: 11838953 [PubMed - indexed for MEDLINE] 1892: Cell Mol Biol (Noisy-le-grand). 2001 Dec;47(8):1319-28. Genetically modified plants: the stakes. Riba G, Chupeau Y. Scientific Direction Plant and Plant Products, INRA, Paris, France. guy.riba@paris.inra.fr Generically modified plants (GMP) are massively used on the American continent in Australia and in China, since they represent an unquestionable potential for progress. New attributes are therefore devoted to the human and animal diet, to the facilitating of culture management, to the reducing of the chemical fertilizer and pesticide usage, and to the conquest of new cultural spaces. Considering itself to be flawed by a too hasty plunge into the market, concomitant with sagging evaluations of other innovations, Europe is confronted by a strong societal debate which blocks GMP cultures and orientates the research towards an evaluation of the environmental and public health risks and an evaluation of their economical and sociological impacts. The authors encourage this societal debate in order to arbitrate the presence of transgenes in conventional productions and products, to define the accepted rules of responsibility, to decide what is not acceptable, and to involve the more upstream actors and operators of the innovation process, all that keeping in mind the agronomical, ecological and economical repercussions of their decisions. Publication Types: Review PMID: 11838952 [PubMed - indexed for MEDLINE] 1893: Eur J Endocrinol. 2002 Feb;146(2):245-9. Effects of pair-feeding and growth hormone treatment on obese transgenic rats. Furuhata Y, Hirabayashi K, Yonezawa T, Takahashi M, Nishihara M. Department of Veterinary Physiology, Veterinary Medical Science, University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan. BACKGROUND: It has been shown that GH-deficient subjects tend to have fat accumulation. We have produced human GH (hGH) transgenic rats that exhibit low circulating hGH levels and hyperphagia. These rats are also characterized by severe obesity, hyperinsulinemia and hyperlipidemia. OBJECTIVE: The present study was conducted in order to elucidate how excess caloric intake and impaired GH secretion account for fat accumulation and metabolic abnormalities in the transgenic rats. DESIGN AND METHODS: The transgenic rats were subjected to either pair-feeding with non-transgenic controls or hGH treatment from 4 to 12 weeks of age, and the effects on fat accumulation and some metabolic parameters were assessed. RESULTS: At the age of 12 weeks, body weight and food intake were greater in transgenic than in control rats by 10% and 27% respectively. The ratio of epididymal white adipose tissue weight to body weight (WAT/BW) was more than three times greater in transgenic than in control rats. Although pair-feeding for 8 weeks decreased body weight, it did not affect the WAT/BW ratio. Treatment with hGH affected neither body weight nor food intake, while it reduced the WAT/BW ratio by 30%. Serum concentrations of triglyceride, free fatty acid, insulin and leptin were all significantly higher in the transgenic than in the control rats. Pair-feeding decreased serum triglyceride, insulin and leptin levels, but not serum free fatty acid levels. On the other hand, hGH treatment decreased only serum leptin concentrations. CONCLUSIONS: These results suggest that severe fat accumulation in the transgenic rats mainly resulted from the decreased lipolytic action of GH, while metabolic abnormalities mainly resulted from excess caloric intake. Publication Types: Research Support, Non-U.S. Gov't PMID: 11834436 [PubMed - indexed for MEDLINE] 1894: BMC Biotechnol. 2002;2:1. Epub 2002 Jan 11. Optimisation of transgene action at the post-transcriptional level: high quality parthenocarpic fruits in industrial tomatoes. Pandolfini T, Rotino GL, Camerini S, Defez R, Spena A. Dipartimento Scientifico Tecnologico, University of Verona, Verona, Italy. pandolfini@sci.univr.it BACKGROUND: Genetic engineering of parthenocarpy confers to horticultural plants the ability to produce fruits under environmental conditions that curtail fruit productivity and quality. The DefH9-iaaM transgene, whose predicted action is to confer auxin synthesis specifically in the placenta, ovules and derived tissues, has been shown to confer parthenocarpy to several plant species (tobacco, eggplant, tomato) and varieties. RESULTS: UC82 tomato plants, a typical cultivar used by the processing industry, transgenic for the DefH9-iaaM gene produce parthenocarpic fruits that are malformed. UC82 plants transgenic for the DefH9-RI-iaaM, a DefH9-iaaM derivative gene modified in its 5'ULR by replacing 53 nucleotides immediately upstream of the AUG initiation codon with an 87 nucleotides-long sequence derived from the rolA intron sequence, produce parthenocarpic fruits of high quality. In an in vitro translation system, the iaaM mRNA, modified in its 5'ULR is translated 3-4 times less efficiently than the original transcript. An optimal expressivity of parthenocarpy correlates with a reduced transgene mRNA steady state level in DefH9-RI-iaaM flower buds in comparison to DefH9-iaaM flower buds. Consistent with the known function of the iaaM gene, flower buds transgenic for the DefH9-RI-iaaM gene contain ten times more IAA than control untransformed flower buds, but five times less than DefH9-iaaM flower buds. CONCLUSIONS: By using an auxin biosynthesis transgene downregulated at the post-transcriptional level, an optimal expressivity of parthenocarpy has been achieved in a genetic background not suitable for the original transgene. Thus, the method allows the generation of a wider range of expressivity of the desired trait in transgenic plants. Publication Types: Comparative Study Research Support, Non-U.S. Gov't PMID: 11818033 [PubMed - indexed for MEDLINE] 1895: Shokuhin Eiseigaku Zasshi. 2001 Aug;42(4):231-6. [Detection of recombinant DNA from genetically modified papaya] [Article in Japanese] Goda Y, Asano T, Shibuya M, Hino A, Toyoda M. National Institute of Health Sciences: 1-18-1, Kamiyoga, Setagaya-ku, Tokyo 158-8501, Japan. A method using polymerase chain reaction (PCR) was developed to detect the genetically modified (GM) papaya (55-1 line), of which the mandatory safety assessment has not been finished in Japan because of insufficient data. The papaya intrinsic papain gene was used as an internal control. The results of PCR amplification of the papain gene segment indicated that a commercial silica membrane type kit (QIAGEN DNeasy plant mini) was useful for extraction of DNA from papaya fruit, but not for extraction from canned papaya fruit. On the other hand, a commercial ion-exchange type kit (QIAGEN Genomic-tip) provided enough purified DNA for PCR from canned papaya fruit. Compared with the parental line and other commercial non-GM papayas, the DNA from GM papaya fruit provided specific amplification bands in PCR with five primer pairs (Nos. 2-6) including beta-glucuronidase and neomycin phosphotransferase II gene-specific ones. On the other hand, the primer pairs recognizing these genes showed false-positive results when we used DNAs from canned papaya. Therefore, we recommend that the primer pairs (Nos. 5 and 6) recognizing the sequences derived from two different species of organism should be used in order to detect specifically the GM papaya in canned fruits. Publication Types: English Abstract Research Support, Non-U.S. Gov't PMID: 11817137 [PubMed - indexed for MEDLINE] 1896: Peptides. 2002 Jan;23(1):117-25. Pharmacological characterization of the nociceptin receptor which mediates reduction of alcohol drinking in rats. Ciccocioppo R, Polidori C, Antonelli L, Salvadori S, Guerrini R, Massi M. Department of Pharmacological Sciences and Experimental Medicine, University of Camerino, 62032, Camerino, Italy. r.ciccocioppo@ca Chronic intracerebroventricular (ICV) treatment with nociceptin/orphanin FQ (NC), the endogenous ligand for the opioid receptor-like 1 (ORL1) receptor, reduces ethanol intake in alcohol-preferring rats and abolishes the rewarding properties of ethanol in the place conditioning paradigm. To pharmacologically characterize the receptor involved, the present study evaluated the effect on ethanol drinking in genetically selected Marchigian Sardinian alcohol-preferring (msP) rats of ICV injections for 8 days of NC or of the NC analogs NC(1-17)NH(2), NC(1-13)NH(2), NC(1-12)NH(2) and [Nphe(1)]NC(1-13)NH(2). In vitro studies indicate that NC, NC(1-17)NH(2), NC(1-13)NH(2) and NC(1-12)NH(2) are agonists, while [Nphe(1)]NC(1-13)NH(2) is a selective antagonist at the ORL1 receptor. Freely feeding and drinking rats were offered 10% ethanol 30 min/day at the beginning of the dark phase of the light cycle. NC significantly attenuated ethanol intake at 500 or 1000 ng/rat (210 or 420 pmol/rat). NC(1-17)NH(2), markedly reduced ethanol intake, but its effect was statistically significant at 1000 (420 pmol/rat), not at 500 ng/rat (210 pmol/rat). After the end of treatment ethanol drinking promptly came back to baseline level. Ethanol consumption was also reduced by NC(1-13)NH(2); however, its effect was less potent and pronounced. NC(1-12)NH(2) did not modify ethanol intake at doses up to 4000 ng/rat (2339 pmol/rat). Water and food consumption were not modified. Treatment with [Nphe(1)]NC(1-13)NH(2), 66 or 99 microg/rat, did not modify ethanol intake; however, [Nphe(1)]NC(1-13)NH(2), 66 microg/rat, given just before 1000 ng/rat of NC(1-17)NH(2), abolished the effect of the agonist. The present results show that the 13 amino acid N-terminal sequence of NC is essential for the effect on ethanol intake and indicate that [Nphe(1)]NC(1-13)NH(2) acts as an antagonist to block the effect of NC. These findings provide further evidence that selective agonists at the ORL-1 receptor attenuate ethanol intake in alcohol-preferring rats and suggest that the NC/ORL1 system may represent an interesting target for treatment of alcohol abuse. Publication Types: Research Support, Non-U.S. Gov't PMID: 11814626 [PubMed - indexed for MEDLINE] 1897: Environ Sci Technol. 2002 Jan 1;36(1):8A. Comment on: Environ Sci Technol. 2001 Dec 1;35(23):472A-473A. Transgenes in native Mexican maize--still no need for concern. Martínez-Soriano J, Bailey AM, Lara-Reyna J, Leal-Klevezas DS. Publication Types: Comment Letter PMID: 11811496 [PubMed - indexed for MEDLINE] 1898: Nature. 2002 Jan 24;415(6870):353. UN attempts to boost biosafety in developing world. Adam D. Publication Types: News PMID: 11807509 [PubMed - indexed for MEDLINE] 1899: Food Drug Law J. 1998;53(1):181-91. No "killer tomatoes": easing federal regulation of genetically engineered plants. Beach JE. Hyman, Phelps & McNamara, P.C., Washington, D.C., USA. PMID: 11795332 [PubMed - indexed for MEDLINE] 1900: Metab Eng. 2002 Jan;4(1):12-21. Metabolic engineering of fatty acid biosynthesis in plants. Thelen JJ, Ohlrogge JB. Department of Plant Biology, Michigan State University, East Lansing, Michigan 48824, USA. Fatty acids are the most abundant form of reduced carbon chains available from nature and have diverse uses ranging from food to industrial feedstocks. Plants represent a significant renewable source of fatty acids because many species accumulate them in the form of triacylglycerol as major storage components in seeds. With the advent of plant transformation technology, metabolic engineering of oilseed fatty acids has become possible and transgenic plant oils represent some of the first successes in design of modified plant products. Directed gene down-regulation strategies have enabled the specific tailoring of common fatty acids in several oilseed crops. In addition, transfer of novel fatty acid biosynthetic genes from noncommercial plants has allowed the production of novel oil compositions in oilseed crops. These and future endeavors aim to produce seeds higher in oil content as well as new oils that are more stable, are healthier for humans, and can serve as a renewable source of industrial commodities. Large-scale new industrial uses of engineered plant oils are on the horizon but will require a better understanding of factors that limit the accumulation of unusual fatty acid structures in seeds. Publication Types: Review PMID: 11800570 [PubMed - indexed for MEDLINE] 1901: Metab Eng. 2002 Jan;4(1):3-11. Metabolic engineering of amino acids and storage proteins in plants. Galili G, Höfgen R. Department of Plant Sciences, The Weizmann Institute of Science, Rehovot 76100, Israel. gad.galili@weizmann.ac.il Amino acid pathways are important targets for plant metabolic engineering. Since plants represent the major global food supply, large efforts are devoted to increasing the content of "essential" amino acids, which are absolutely required in human foods and animal feeds. Engineering of amino acids is also undertaken to improve plant growth and stress tolerance. Many of the pathways of amino acid metabolism in plants have been elucidated, and genes encoding most of the enzymes are now available. The expression of recombinant genes in transgenic plants, coupled with genetic and biochemical approaches, has contributed significantly to the understanding of regulatory networks of the metabolism of amino acids and their incorporation into proteins. This knowledge is now being extensively applied to metabolic engineering of crops, and this is reflected by a large patent literature. The problems of engineering plant amino acid metabolism, and ways to solve them, are discussed using the essential amino acids lysine and methionine as examples. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 11800569 [PubMed - indexed for MEDLINE] 1902: J Allergy Clin Immunol. 2002 Jan;109(1):143-9. Linear IgE epitope mapping of the English walnut (Juglans regia) major food allergen, Jug r 1. Robotham JM, Teuber SS, Sathe SK, Roux KH. Department of Biological Science and Structural Biology Program, Florida State University, Tallahassee 32306-4370, USA. BACKGROUND: Peanut and tree nut allergies can be life-threatening, and they appear to be growing in prevalence. Jug r 1, a 2S albumin seed storage protein, was previously characterized as a major English walnut food allergen. OBJECTIVE: We sought to identify the linear IgE-binding epitopes of Jug r 1 and to determine which, if any, amino acids are necessary for this binding to occur. METHODS: Pools of sera from walnut-allergic patients and overlapping peptides synthesized on an activated cellulose membrane were used to screen for IgE-binding epitopes. Mutational analysis of the immunodominant epitope was carried out through single and multisite amino acid substitutions. Inhibition assays were performed through use of affinity-purified IgE, soluble forms of the epitope peptide, and the recombinant 2S albumin, rJug r 1. RESULTS: One immunodominant linear epitope was identified. Amino acid mutations to the epitope demonstrated that the residues RGEE, at positions 36 through 39, were minimally required for IgE binding. Probing of this epitope with sera from each of 20 patients revealed 15 of the sera to be positive. Binding of patients' IgE to the epitope was inhibited with a soluble form of the peptide; however, soluble peptide did not completely inhibit the binding of IgE to the intact rJug r 1. CONCLUSION: One major linear IgE-reactive epitope and its critical core amino acid residues have been identified. Mutation of any of these core amino acids resulted in loss of IgE binding to the epitope, and this points toward the feasibility of reducing allergenicity in genetically modified walnuts. However, strong evidence for the existence of conformational epitopes was also obtained. Publication Types: Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. PMID: 11799381 [PubMed - indexed for MEDLINE] 1903: J Virol. 2002 Feb;76(4):1999-2002. Human CD59 incorporation into porcine endogenous retrovirus particles: implications for the use of transgenic pigs for xenotransplantation. Takefman DM, Spear GT, Saifuddin M, Wilson CA. Division of Cellular and Gene Therapies, Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, Maryland 20892, USA. Transgenic pigs have been engineered to express human CD59 (hCD59) in order to suppress hyperacute rejection of xenotransplants in human recipients. In this study, porcine endogenous retrovirus (PERV) was produced in a porcine cell line expressing hCD59 in order to examine the effect of this complement control protein on PERV neutralization by human sera. hCD59 was found to be incorporated into PERV particles produced from engineered ST-IOWA cells. PERV incorporation of hCD59 resulted in a dramatic inhibition of complement-mediated virolysis by human serum. However, incorporation of hCD59 had no effect on neutralization of PERV by human serum, as measured in infectivity assays. Our results suggest that the use of organs from hCD59 transgenic pigs will inhibit complement-mediated virolysis, but will not compromise the protective effects of human sera on the neutralization of PERV particles. PMID: 11799196 [PubMed - indexed for MEDLINE] 1904: Ann Nutr Metab. 2001;45(6):235-54. Safety considerations of DNA in food. Jonas DA, Elmadfa I, Engel KH, Heller KJ, Kozianowski G, König A, Müller D, Narbonne JF, Wackernagel W, Kleiner J. Institute of Nutritional Sciences, University of Vienna, Vienna, Austria. Recombinant DNA techniques are capable of introducing genetic changes into food organisms that are more predictable than those introduced through conventional breeding techniques. This review discusses whether the consumption of DNA in approved novel foods and novel food ingredients derived from genetically modified organisms (GMOs) can be regarded as being as safe as the consumption of DNA in existing foods. It concludes that DNA from GMOs is equivalent to DNA from existing food organisms that has always been consumed with human diets. Any risks associated with the consumption of DNA will remain, irrespective of its origin, because the body handles all DNA in the same way. The breakdown of DNA during food processing and passage through the gastrointestinal tract reduces the likelihood that intact genes capable of encoding foreign proteins will be transferred to gut microflora. The review does not specifically address food safety issues arising from the consumption of viable genetically modified microorganisms but it shows that the likelihood of transfer and functional integration of DNA from ingested food by gut microflora and/or human cells is minimal. Information reviewed does not indicate any safety concerns associated with the ingestion of DNA per se from GMOs resulting from the use of currently available recombinant DNA techniques in the food chain. Copyright 2001 S. Karger AG, Basel Publication Types: Review PMID: 11786646 [PubMed - indexed for MEDLINE] 1905: Bioelectrochemistry. 2002 Jan;55(1-2):107-12. Recent biotechnological developments of electropulsation. A prospective review. Teissié J, Eynard N, Vernhes MC, Bénichou A, Ganeva V, Galutzov B, Cabanes PA. IPBS CNRS (UMR 5089), 205 Route de Narbonne, 31077 Toulouse Cedex, France. justin@ipbs.fr During the last 25 years, basic research has improved our knowledge on the molecular mechanisms triggered at the membrane level by electric pulses. Applied aspects may now be used under safe conditions. Electropulsation is known as a very efficient tool for obtaining gene transfer in many species to produce genetically modified organisms (GMO). This is routinely used for industrial purposes to transfer exogenous activities in bacteria, yeasts and plants. The method is simple and of a low cost. But electropulsation is not limited to this application for biotechnological purposes. It is known that the field-associated membrane alterations can be irreversible. The pulsed species cannot recover after the treatment. Their viability is strongly affected. This appears as a very promising technology for the eradication of pathogenic microorganisms. Recent developments are proposed for sterilization purposes. New flow technologies of field generation allow the treatment of large volumes of solution. When high flow rates are used, microorganisms are submitted both to a hydromechanical and to an electrical stress. The synergy of the two effects may be present when suitable pulsing conditions are chosen. Several examples for the treatment of domestic water and in the food industry are described. Walled microorganisms are affected not only at the membrane level. We observed that alterations are present on the cell wall. A very promising technology is the associated controlled leakage of the cytoplasmic soluble proteins. Large dimeric proteins such as beta-galactosidases can be extracted at a high yield. High volumes can be treated by using a flow process. Extraction of proteins is obtained with many systems including mammalian cells. Publication Types: Review PMID: 11786352 [PubMed - indexed for MEDLINE] 1906: Cell Mol Neurobiol. 2001 Aug;21(4):389-401. A potential role of central insulin in learning and memory related to feeding. Gerozissis K, Rouch C, Lemierre S, Nicolaidis S, Orosco M. Laboratoire de Physiopathologie de la Nutrition, Université Paris 7, France. gerozi@paris7.jussieu.fr 1. Hypothalamic insulin (HI) is well known for its role in feeding regulation. In addition, its concentration is modified in response to meals. Recent studies suggest that brain insulin participates in memory processes, possibly through stimulation by glucose. 2. The present microdialysis study focused on local in vivo regulation of HI by glucose and on the effects of aging on HI, since aging is characterized by deterioration of memory, body weight regulation, and central glucose utilization. Glucose (8 mM) infused for 5 min increased extracellular HI levels rapidly, by 4.6-fold, and cerebellar insulin levels by 0.4-fold only, suggesting a specific area-dependent regulation of HI by glucose. Neither insulinemia nor glycemia were affected, suggesting a central mechanism. The same dose of glucose induced a modest (0.4-fold), delayed (45 min) increase in hypothalamic serotonin, suggesting that the effect of glucose on HI is independent of a previously defined local serotonin-induced insulin release. HI levels in old normal weight rats were half the levels of young rats. In genetically old obese (fa/fa) Zucker rats, HI concentration was 30% of that in young normal rats, suggesting a deterioration of HI availability when aging and obesity are combined. 3. The above results, in line with recent considerations on a potential role of central insulin in learning and memory, suggest particular effects of HI on feeding and memory and probably on a specific "memory for food." Publication Types: Comparative Study PMID: 11775068 [PubMed - indexed for MEDLINE] 1907: Adv Exp Med Biol. 2001;495:299-303. Oral immunization of human with transgenic lettuce expressing hepatitis B surface antigen. Kapusta J, Modelska A, Pniewski T, Figlerowicz M, Jankowski K, Lisowa O, Plucienniczak A, Koprowski H, Legocki AB. Institute of Bioorganic Chemistry, Polish Academy of Sciences, Noskowskiego 12, 61-704 Poznań, Poland. Publication Types: Clinical Trial Research Support, Non-U.S. Gov't PMID: 11774582 [PubMed - indexed for MEDLINE] 1908: Pharmacogenetics. 2002 Jan;12(1):55-65. Implications of circadian gene expression in kidney, liver and the effects of fasting on pharmacogenomic studies. Kita Y, Shiozawa M, Jin W, Majewski RR, Besharse JC, Greene AS, Jacob HJ. Department of Physiology, Medical College of Wisconsin, Milwaukee, WI 53228, USA. Pharmacogenomics offers the potential to define metabolic pathways and to provide increased knowledge of drug actions. We studied relative levels of gene expression in the rat using a microarray with 8448 rat UniGenes (1928 known genes, 6520 unknown ESTs) in the liver and kidney as a function of time of day and then of feeding regime, which are common variables in preclinical pharmacogenomic studies. We identified 597 genes, including several key metabolic pathways, whose relative expression levels are significantly affected by time of day: expression of some was further modified by feeding state. These would have sparked interest in a pharmacogenomic study. Our study demonstrates that two common variables in pharmacogenomic studies can have dramatic effects on gene expression. This study provides investigators with baseline information for both kidney and liver with respect to 'normal' changes in gene expression influenced by time of day and feeding state. It also identifies 18 new genes that should be investigated for a role in circadian rhythms in peripheral tissues. Publication Types: Research Support, U.S. Gov't, P.H.S. PMID: 11773865 [PubMed - indexed for MEDLINE] 1909: Ying Yong Sheng Tai Xue Bao. 2000 Dec;11(6):856-60. [Lifetable of cotton bollworm under different control condition] [Article in Chinese] Yang Y, Pang X, Liang G. Laboratory Insect Ecology, South China Agricultural University, Guangzhou 510642. The lifetables of 3rd and 4th generation of cotton bollworm Helicoverpa armigera Hubner were established by action factors under four different control conditions. The results showed that under natural control, the egg parasitic rates of 3rd and 4th cotton bollworm were 4.39% and 1.19%, the predatory rates were 21.11% and 15.80%, and the larvae parasitic rates were 19.34% and 16.20% respectively. The indexes of population trend were only 0.0032 and 0.0014 respectively, after using selective pesticides IKI and Bt to control cotton bollworm. Though the index of population trend decreased to 48.66% and 61.67% after contantly praying chemical pesticides, the number of predators also decreased to 23.48% and 41.26% respectively. Transgenic Bt cotton could inhibit the population increase of cotton bollworm, but had no effect on predators. The parasitic rates of 3rd and 4th cotton bollworm on Transgenic Bt cotton were 51.14% and 0% respectively, as compared to control. Publication Types: English Abstract PMID: 11767558 [PubMed - indexed for MEDLINE] 1910: J AOAC Int. 2001 Nov-Dec;84(6):1941-6. Sampling grain shipments to detect genetically modified seed. Whitaker TB, Freese L, Giesbrecht FG, Slate AB. USDA/ARS, North Carolina State University, Raleigh 27695-7625, USA. Tom_Whitaker@ncsu.edu Using the binomial distribution, the effect of sample size on the variability among sample test results when sampling a lot with 1.0% genetically modified (GM) or biotech seed was evaluated. The coefficient of variation, cv, among 500-seed sample test results taken from a lot with truly 1.0% was computed to be 44.5%. Increasing sample size to 1000 seeds reduced the cv among sample test results to 31.5%. The effects of sample size and accept/reject limits on the buyer's risk (bad lots accepted) and the seller's risk (good lots rejected) was also evaluated assuming a tolerance of 1.0% GM seed. Increasing sample size decreases both the buyer's and seller's risks at the same time. Using an accept/reject limit below the regulatory tolerance decreases the buyer's risk, but increases the seller's risk. Using an accept/reject limit above the regulatory tolerance decreases the seller's risk but increases the buyer's risk. PMID: 11767166 [PubMed - indexed for MEDLINE] 1911: J AOAC Int. 2001 Nov-Dec;84(6):1891-901. Determination of Cry9C protein in corn-based foods by enzyme-linked immunosorbent assay: interlaboratory study. Trucksess NW. U.S. Food and Drug Administration, Center for Food Safety and Applied Nutrition, Washington, DC 20204, USA. mtruckse@cfsan.fda.gov The performance of a commercially available enzyme-linked immunosorbent assay kit (Enviro-Logix) was assessed for the determination of Cry9C protein, which is produced by the genetically modified corn StarLink, in 8 types of corn-based foods (starch, refined oil, soft tortillas, tortilla chips, corn flakes, corn puffs, corn muffins, and corn bread) in an interlaboratory study involving 7 laboratories in the United States. The assay kit is a double antibody sandwich and is based on the specific interaction between antibody and antigen. The Cry9C protein analyte is sandwiched between 2 antibodies, one to capture the analyte and the other is conjugated to the enzyme, horseradish peroxidase. The enzyme uses tetramethylbenzidine/peroxide for color development. A strong acid stopping reagent is then used to change the color from blue to a stable yellow. The intensity of the color is proportional to the concentration of the Cry9C protein. In this study blind duplicates of control samples (blank material prepared from non- StarLink corn), spiked samples (blank material with the addition of Cry9C protein), and samples containing incurred analyte (products prepared with StarLink corn) were analyzed. Cry9C protein from 2 different sources was used to spike the food products. Cry9C protein produced and purified from a bacterial host was used to prepare spiked test samples at 2.72 and 6.8 ng/g. Cry9C protein from StarLink corn flour was used to prepare spiked samples at 1.97 ng/g. Average recoveries for samples spiked with corn flour Cry9C protein at 1.97 ng/g ranged from 73 to 122%, within-laboratory relative standard deviations (RSDr) ranged from 6 to 22%, and between-laboratories relative standard deviations (RSDR) ranged from 16 to 56%. Average recoveries for samples spiked with bacterial Cry9C protein at 2.72 and 6.8 ng/g ranged from 27 to 96% and from 32 to 113%, respectively; RSDr values ranged from 10 to 35% and from 7 to 38%, respectively; and the RSDR ranged from 28 to 84% and 15 to 75%, respectively. The incurred test samples were found to contain Cry9C protein at levels ranging from 0.8 to 3187 ng/g depending on the product, RSDr values ranged from 5 to 16% and RSDR values ranged from 11 to 71%. Results of the statistical analysis indicate that this method is applicable to the determination of Cry9C protein in the 8 types of collaboratively studied corn-based products containing Cry9C protein (from StarLink) at levels of > or =2 ng/g. PMID: 11767159 [PubMed - indexed for MEDLINE] 1912: J AOAC Int. 2001 Nov-Dec;84(6):1855-64. Validation of PCR methods for quantitation of genetically modified plants in food. Hübner P, Waiblinger HU, Pietsch K, Brodmann P. Kantonales Labor Zürich, Switzerland. For enforcement of the recently introduced labeling threshold for genetically modified organisms (GMOs) in food ingredients, quantitative detection methods such as quantitative competitive (QC-PCR) and real-time PCR are applied by official food control laboratories. The experiences of 3 European food control laboratories in validating such methods were compared to describe realistic performance characteristics of quantitative PCR detection methods. The limit of quantitation (LOQ) of GMO-specific, real-time PCR was experimentally determined to reach 30-50 target molecules, which is close to theoretical prediction. Starting PCR with 200 ng genomic plant DNA, the LOQ depends primarily on the genome size of the target plant and ranges from 0.02% for rice to 0.7% for wheat. The precision of quantitative PCR detection methods, expressed as relative standard deviation (RSD), varied from 10 to 30%. Using Bt176 corn containing test samples and applying Bt176 specific QC-PCR, mean values deviated from true values by -7to 18%, with an average of 2+/-10%. Ruggedness of real-time PCR detection methods was assessed in an interlaboratory study analyzing commercial, homogeneous food samples. Roundup Ready soybean DNA contents were determined in the range of 0.3 to 36%, relative to soybean DNA, with RSDs of about 25%. Taking the precision of quantitative PCR detection methods into account, suitable sample plans and sample sizes for GMO analysis are suggested. Because quantitative GMO detection methods measure GMO contents of samples in relation to reference material (calibrants), high priority must be given to international agreements and standardization on certified reference materials. Publication Types: Validation Studies PMID: 11767156 [PubMed - indexed for MEDLINE] 1913: Dev Biol (Basel). 2001;106:123-31; discussion 131-2, 143-60. Transgenic animal models that might be useful in identifying unsuspected oncogenic factors in tumour cell substrates. Sistare FD. Center for Drug Evaluation and Research, Food and Drug Administration, Laurel, MD 20708, USA. The use of alternative short term models for the assessment of pharmaceutical carcinogenicity potential is expanding and evolving. Several mouse models of accelerated carcinogenesis have gained user confidence and are being applied to product development decision-making. These models have been highlighted as part of a consortium effort organized under ILSI to further evaluate their strengths and limitations compared to the standard two-years, two species cancer bioassay. Examples of trends that are developing in our understanding of applicability of these models to chemical carcinogenesis have been presented but a more comprehensive review of all the currently available data has been avoided. Such a review would be soon outdated as the anticipated data from the ILSI consortium studies will soon become available and greatly further impact our present appreciation of the capabilities of these models. As strengths and limitations of those models are being elucidated, mechanistic understanding of the models is expanding. In addition to those reviewed briefly in this paper, numerous other transgenic models with micro-injected oncogenes and disrupted tumour suppressor genes have demonstrated enhanced propensity for developing cancer. Critical experiments can be designed to test, conceptually, for the ability of any of these models to detect tumorigenic potential for a given dose of adventitious agent, or residual DNA sequence. The choice of model and of DNA sequence or adventitious agent to begin this evaluation must be carefully considered. From such data, risk may be better identified or prioritized, but such approaches may not be practical as general routine screens of finished vaccine product with unknown residual DNA sequence or adventitious agents. PMID: 11761226 [PubMed - indexed for MEDLINE] 1914: Food Addit Contam. 2001 Dec;18(12):1135-40. Contribution of European research to risk analysis. Boenke A. European Commission, DG Research, Directorate E-Life Sciences, Unit 2-Health, Food and Environment, Brussels, Belgium. Achim.Boenke@cec.eu.int The European Commission's, Quality of Life Research Programme, Key Action 1-Health, Food & Nutrition is mission-oriented and aims, amongst other things, at providing a healthy, safe and high-quality food supply leading to reinforced consumer confidence in the safety, of European food. Its objectives also include the enhancing of the competitiveness of the European food supply. Key Action 1 is currently supporting a number of different types of European collaborative projects in the area of risk analysis. The objectives of these projects range from the development and validation of prevention strategies including the reduction of consumers risks; development and validation of new modelling approaches, harmonization of risk assessment principles methodologies and terminology; standardization of methods and systems used for the safety evaluation of transgenic food; providing of tools for the evaluation of human viral contamination of shellfish and quality control; new methodologies for assessing the potential of unintended effects of genetically modified (genetically modified) foods; development of a risk assessment model for Cryptosporidium parvum related to the food and water industries, to the development of a communication platform for genetically modified organism, producers, retailers, regulatory authorities and consumer groups to improve safety assessment procedures, risk management strategies and risk communication; development and validation of new methods for safety testing of transgenic food; evaluation of the safety and efficacy of iron supplementation in pregnant women, evaluation of the potential cancer-preventing activity of pro- and pre-biotic ('synbiotic') combinations in human volunteers. An overview of these projects is presented here. PMID: 11761126 [PubMed - indexed for MEDLINE] 1915: Food Addit Contam. 2001 Dec;18(12):1130-4. Risk communication--the perceptions and realities. Trautman TD. General Mills, Minneapolis, Minnesota 55426-1350, USA. tom.trautman@genmills.com This paper defines risk communication and puts it into the perspective of risk analysis as a whole. Case studies originating for the food industry are described in the areas of food colours, pesticides and genetically, modified foods to exemplify both the difficulties of risk communication and the lessons that can be learned. This paper concludes by suggesting ways in which successful risk communication should be managed. PMID: 11761125 [PubMed - indexed for MEDLINE] 1916: Food Addit Contam. 2001 Dec;18(12):1099-107. Food allergy--towards predictive testing for novel foods. Oehlschlager S, Reece P, Brown A, Hughson E, Hird H, Chisholm J, Atkinson H, Meredith C, Pumphrey R, Wilson P, Sunderland J. Central Science Laboratory, Sand Hutton, York, UK. s.oehlschlager@csl.gov.uk The risks associated with IgE-mediated food allergy highlight the need for methods to screen for potential food allergens. Clinical and immunological tests are available for the diagnosis of food allergy to known food allergens, but this does not extend to the evaluation, or prediction of allergenicity in novel foods. This category, includes foods produced using novel processes genetically modified (GM) foods, and foods that might be used as alternatives to traditional foods. Through the collation and analysis of the protein sequences of known allergens and their epitopes, it is possible to identify related groups which correlate with observed clinical cross-reactivities. 3-D modelling extends the use of sequence data and can be used to display eptiopes on the surface of a molecule. Experimental models support sequence analysis and 3-D modelling. Observed cross-reactivities can be examined by Western blots prepared from native 2-D gels of a whole food preparation (e.g. hazelnut, peanut), and common proteins identified. IgEs to novel proteins can be raised in Brown Norway rat (a high IgE responder strain) and the proteins tested in simulated digest to determine epitope stability. Using the CSL serum bank, epitope binding can be examined through the ability of an allergen to cross-link the high affinity IgE receptor and thereby release mediators using in vitro cell-based models. This range of methods, in combination with data mining, provides a variety of screening options for testing the potential of a novel food to be allergenic, which does not involve prior exposure to the consumer. Publication Types: Research Support, Non-U.S. Gov't PMID: 11761121 [PubMed - indexed for MEDLINE] 1917: Nutr Cancer. 2001;39(2):259-66. Modulation of 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine-induced mutation in the cecum and colon of big blue rats by conjugated linoleic acid and 1,2-dithiole-3-thione. Yang H, Stuart GR, Glickman BW, de Boer JG. Centre for Environmental Health, University of Victoria, Victoria, BC, Canada V8W 3N5. 2-Amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) is a potent mutagen and suspected human carcinogen present in cooked protein-rich food. It preferentially induced colon tumors in male rats and mammary tumors in female rats. In the present study, the in vivo antimutagenic efficacy of two dietary compounds, conjugated linoleic acid (CLA) and 1,2-dithiole-3-thione (DTT), against PhIP was explored using 1acI transgenic Big Blue rats. Five- or six-week-old male Big Blue rats were fed a diet containing CLA (0.5%, wt/wt) or DTT (0.005%, wt/wt) starting one week before exposure to 200 ppm PhIP for 61 days. PhIP treatment induced a approximately 8- to 16-fold increase in the mutation frequency (MF) in the colon. The induced MF was significantly lower in the cecum than in the proximal and distal colon (approximately 52 x 10(-5) vs. 100 x 10(-5), p < 0.008). CLA and DTT significantly reduced the PhIP-induced MF in the distal colon (p < 0.05) by 14% and 24%, respectively. The frequency of -1 frameshift mutations was lower in the distal colon of CLA- or DTT-treated rats. This protective effect was not observed in the cecum or in the proximal colon. In contrast, the PhIP-induced MF in the cecum (specifically, the frequency of -1 frameshifts and GC-->TA transversions) was elevated by 43% after treatment with CLA. In conclusion, CLA and DTT modulate PhIP-induced mutagenesis in a tissue-specific manner, and different modulation pathways are employed by CLA and DTT. PMID: 11759290 [PubMed - indexed for MEDLINE] 1918: J Pharmacol Exp Ther. 2002 Jan;300(1):91-6. Erratum in: J Pharmacol Exp Ther 2002 Mar;300(3):1131. Effects of pharmacological agents upon a transgenic model of Parkinson's disease in Drosophila melanogaster. Pendleton RG, Parvez F, Sayed M, Hillman R. Temple University, Philadelphia, Pennsylvania, USA. rgp@mymailstation.com The human gene that codes for the protein alpha-synuclein has been transferred into the Drosophila melanogaster genome. The transgenic flies recapitulate some of the essential features of Parkinson's disease. These include the degeneration of certain dopaminergic neurons in the brain accompanied by the appearance of age-dependent abnormalities in locomotor activity. In the present study, we tested the locomotor response of these transgenic flies to prototypes of the major classes of drugs currently used to treat this disorder. A time course study was first conducted to determine when impaired locomotor activity appeared relative to normal "wild-type" flies. A climbing or negative geotaxis assay measuring the ability of the organisms to climb up the walls of a plastic vial was used. Based on the results obtained, normal and transgenic flies were treated with each of the drugs in their food for 13 days and then assayed. The activity of transgenic flies treated with L-DOPA was restored to normal. Similarly, the dopamine agonists pergolide, bromocriptine, and 2,3,4,5-tetrahydro-7,8-dihydroxy- 1-phenyl-1H-3-benzazepine (SK&F 38393) were substantially effective. Atropine, the prototypical muscarinic cholinergic receptor antagonist, was also effective but to a lesser extent than the other antiparkinson compounds. p-Chlorophenylalanine, an inhibitor of serotonin synthesis, was without beneficial effect as was alpha-methyl-p-tyrosine, an inhibitor of tyrosine hydroxylase, the rate-limiting step in catecholamine biosynthesis. This behavioral study further demonstrates the utility of this model in studying Parkinson's disease and reinforces the concept that inhibition of the action of alpha-synuclein may be useful in its treatment as may dopamine D(1) receptor agonists. Publication Types: Research Support, U.S. Gov't, P.H.S. PMID: 11752102 [PubMed - indexed for MEDLINE] 1919: Lancet. 2001 Dec 8;358(9297):1970. African biotechnologists debate GM foods. Wendo C. Publication Types: News PMID: 11747931 [PubMed - indexed for MEDLINE] 1920: Plant Physiol. 2001 Dec;127(4):1383-9. Floral fragrance. New inroads into an old commodity. Vainstein A, Lewinsohn E, Pichersky E, Weiss D. The Kennedy Leigh Centre for Horticultural Research, Faculty of Agriculture, Food and Environmental Quality Sciences, The Hebrew University of Jerusalem, Rehovet 76100, Israel. vain@agri.huji.ac.il Publication Types: Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Review PMID: 11743078 [PubMed - indexed for MEDLINE] 1921: Nature. 2001 Nov 29;414(6863):541-3. Comment in: Nature. 2002 Apr 11;416(6881):601-2; discussion 600, 602. Nature. 2002 Apr 11;416(6881):602. Nature. 2002 Jun 27;417(6892):897; author reply 897-8. Nature. 2002 Jun 27;417(6892):897; author reply 897-8. Nature. 2002 Oct 24;419(6909):772-6. Nature. 2005 Aug 11;436(7052):760. Transgenic DNA introgressed into traditional maize landraces in Oaxaca, Mexico. Quist D, Chapela IH. Department of Environmental Science, Policy and Management, University of California, Berkeley 94720-3110, USA. Concerns have been raised about the potential effects of transgenic introductions on the genetic diversity of crop landraces and wild relatives in areas of crop origin and diversification, as this diversity is considered essential for global food security. Direct effects on non-target species, and the possibility of unintentionally transferring traits of ecological relevance onto landraces and wild relatives have also been sources of concern. The degree of genetic connectivity between industrial crops and their progenitors in landraces and wild relatives is a principal determinant of the evolutionary history of crops and agroecosystems throughout the world. Recent introductions of transgenic DNA constructs into agricultural fields provide unique markers to measure such connectivity. For these reasons, the detection of transgenic DNA in crop landraces is of critical importance. Here we report the presence of introgressed transgenic DNA constructs in native maize landraces grown in remote mountains in Oaxaca, Mexico, part of the Mesoamerican centre of origin and diversification of this crop. PMID: 11734853 [PubMed - indexed for MEDLINE] 1922: Annu Rev Entomol. 2002;47:845-81. Economic, ecological, food safety, and social consequences of the deployment of bt transgenic plants. Shelton AM, Zhao JZ, Roush RT. Department of Entomology, Cornell University, New York State Agricultural Experiment Station, Geneva, New York 14456, USA. ams5@cornell.edu Transgenic plants expressing insecticidal proteins from the bacterium, Bacillus thuringiensis (Bt), are revolutionizing agriculture. Bt, which had limited use as a foliar insecticide, has become a major insecticide because genes that produce Bt toxins have been engineered into major crops grown on 11.4 million ha worldwide in 2000. Based on the data collected to date, generally these crops have shown positive economic benefits to growers and reduced the use of other insecticides. The potential ecological and human health consequences of Bt plants, including effects on nontarget organisms, food safety, and the development of resistant insect populations, are being compared for Bt plants and alternative insect management strategies. Scientists do not have full knowledge of the risks and benefits of any insect management strategies. Bt plants were deployed with the expectation that the risks would be lower than current or alternative technologies and that the benefits would be greater. Based on the data to date, these expectations seem valid. Publication Types: Review PMID: 11729093 [PubMed - indexed for MEDLINE] 1923: Plant J. 2001 Oct;28(2):181-90. The role of SPY and its TPR domain in the regulation of gibberellin action throughout the life cycle of Petunia hybrida plants. Izhaki A, Swain SM, Tseng TS, Borochov A, Olszewski NE, Weiss D. The Institute of Plant Sciences and Genetics in Agriculture, Faculty of Agricultural, Food and Environmental Quality Sciences, The Hebrew University of Jerusalem, PO Box 12, Rehovot 76100, Israel. SPY acts as a negative regulator of gibberellin (GA) action in Arabidopsis, but its mode of action and regulation are still unknown. SPY over-expression in transgenic petunia plants affected various GA-regulated processes, including seed germination, shoot elongation, flower initiation, flower development and the expression of a GA-induced gene, GIP. A similar phenotype was obtained when wild-type petunia plants were treated with the GA-biosynthesis inhibitor, paclobutrazol. The N-terminus of SPY contains tetratricopeptide repeats (TPR). TPR motifs participate in protein-protein interactions, suggesting that SPY is part of a multiprotein complex. To test this hypothesis, we over-expressed the SPY's TPR region without the catalytic domain in transgenic petunia and generated a dominant-negative SPY mutant. The transgenic seeds were able to germinate on paclobutrazol, suggesting an enhanced GA signal. We cloned the petunia SPY homologue, PhSPY, and showed that its mRNA level is not affected by GA or ABA. The results of this study support the role of SPY as a negative regulator of GA action, suggest that the TPR domain is required for the interaction with other proteins to form an active complex and indicate that different plants use similar mechanisms to transduce the GA signal. Publication Types: Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. PMID: 11722761 [PubMed - indexed for MEDLINE] 1924: Vopr Pitan. 2001;70(5):47-8. [New educational programs for thematic improvement of sanitary doctors in the area of state regulation modified food sources] [Article in Russian] Korolev AA, Elizarova EV. Publication Types: News PMID: 11715701 [PubMed - indexed for MEDLINE] 1925: Qual Assur. 2000 Jan-Mar;8(1):33-6. Controversy over genetically modified organisms: the governing laws and regulations. Keatley KL. Gilead Sciences, 2860 Wilderness Place, Boulder, CO 80301, USA. klkeat@juno.com Genetically Modified Organisms (GMOs) are increasingly becoming a topic of controversy in the U.S. and abroad. The public is questioning their safety and wanting the products labeled as genetically modified. There are other concerns from some of the scientific world and some government officials and organizations such as the Food & Agricultural Organization (FAO) that question whether adequate research has been done to qualify GMOs as safe for long-term use. Of particular concern are the allergenic properties, a GMO may impart, possible transfer effects of antibiotic resistance (given that antibiotic resistant marker genes are used for many GMOs), the expression of previously unexpressed traits, and the drift of pollen from genetically modified crops. It has also been noted that the laws and regulations governing the biotechnology world are outdated, are not comprehensive, and span too many agencies. The primary agencies currently regulating biotechnology are the U.S. Department of Agriculture (USDA), the Food and Drug Administration (FDA), and the Environmental Protection Agency (EPA). PMID: 11710306 [PubMed - indexed for MEDLINE] 1926: Asia Pac J Clin Nutr. 2001;10 Suppl:S4-12. Comment in: Asia Pac J Clin Nutr. 2001;10 Suppl:S1-3. Achieving household nutrition security in societies in transition: an overview. Gopalan C. Nutrition Foundation of India, New Delhi. nfi@ren02.nic.in The achievement of nutrition security at the household level involves adequacy of food supply at the national level and equitable distribution of food among the population in accordance with their physiological needs. The emergence of globalization and market liberalization and the increasing power of some transnational corporations that are advocating pharmaceutical shortcuts have raised concerns in many developing countries. In order to achieve adequacy of food production, earlier mistakes (such as a reliance on unsustainable new technologies) need to be corrected and the resultant imbalances with respect to food production need to be reversed. Emerging new technologies, including genetic modifications, need to be effectively harnessed and adapted with due consideration to safety and sustainability. There is a need to collect convincing evidence of the efficacy and safety of genetically modified foods before they can gain general public acceptance. Information technology will play an important role in future programmes of food production and developing countries must strive to achieve access to this technology. There is considerable scope and need for the expansion of agro-based industries in villages and townships. This could create job opportunities and could also lead to better production and more effective utilization of local food resources by the community and reduce the present considerable loss of perishable food items. Household nutrition security means more than avoidance of chronic starvation. Policy makers of developing countries should set, as their target in the next century, the achievement of adequate nutrition rather than mere survival. Publication Types: Review PMID: 11708581 [PubMed - indexed for MEDLINE] 1927: Plant Physiol. 2001 Nov;127(3):1256-65. Enhanced levels of the aroma and flavor compound S-linalool by metabolic engineering of the terpenoid pathway in tomato fruits. Lewinsohn E, Schalechet F, Wilkinson J, Matsui K, Tadmor Y, Nam KH, Amar O, Lastochkin E, Larkov O, Ravid U, Hiatt W, Gepstein S, Pichersky E. Newe Ya'ar Research Center, Agricultural Research Organization, Ramat Yishay 30095, Israel. twefraim@netvision.net.il The aromas of fruits, vegetables, and flowers are mixtures of volatile metabolites, often present in parts per billion levels or less. We show here that tomato (Lycopersicon esculentum Mill.) plants transgenic for a heterologous Clarkia breweri S-linalool synthase (LIS) gene, under the control of the tomato late-ripening-specific E8 promoter, synthesize and accumulate S-linalool and 8-hydroxylinalool in ripening fruits. Apart from the difference in volatiles, no other phenotypic alterations were noted, including the levels of other terpenoids such as gamma- and alpha-tocopherols, lycopene, beta-carotene, and lutein. Our studies indicate that it is possible to enhance the levels of monoterpenes in ripening fruits by metabolic engineering. Publication Types: Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. PMID: 11706204 [PubMed - indexed for MEDLINE] 1928: Nat Neurosci. 2001 Dec;4(12):1182-6. A Gr receptor is required for response to the sugar trehalose in taste neurons of Drosophila. Dahanukar A, Foster K, van der Goes van Naters WM, Carlson JR. Department of Molecular, Cellular, and Developmental Biology, Yale University, PO Box 208103, New Haven, Connecticut 06520-8103, USA. We recently identified from the Drosophila genome database a large family of G protein-coupled receptor genes, the Gr genes, and predicted that they encode taste receptors on the basis of their structure and specificity of expression. The expression of Gr genes in gustatory neurons has subsequently been confirmed and 56 family members have been reported. Here we provide functional evidence that one Gr gene, Gr5a, encodes a taste receptor required for response to the sugar trehalose. In two different mutants that carry deletions in Gr5a, electrophysiological and behavioral responses to trehalose were diminished but the response to sucrose was unaffected. Transgenic rescue experiments showed that Gr5a confers response to trehalose. The results correlate a particular taste ligand with a Gr receptor and indicate a role for G protein-mediated signaling in the transduction of sweet taste in Drosophila. Publication Types: Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. PMID: 11704765 [PubMed - indexed for MEDLINE] 1929: J Nutr. 2001 Nov;131(11 Suppl):3082S-6S. Protection by dietary compounds against mutation in a transgenic rodent. de Boer JG. Centre for Environmental Health, University of Victoria, Victoria, British Columbia, Canada. jdboer@uvic.ca One of the most relevant biomarkers of genotoxicity and, potentially, carcinogenesis is the occurrence of mutations. Data indicate that carcinogens are highly specific with regard to their target tissue in inducing both tumors and mutations. This specificity may reflect the dependence on tissue-specific metabolic activation, the organ-specific environment or both. Ideally, therefore, mutation should be determined in a real animal rather than in a cell culture system. The lacI transgenic rodent model provides such a system. We have used this model to investigate tissue, species and sex specificity of mutation induced by selected dietary carcinogens and to examine how some compounds may alter the induction of mutation. We have studied mutation using several chemicals, including the dietary heterocyclic amine 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP), the environmentally important aromatic hydrocarbon benzo[a]pyrene and the food contaminant aflatoxin B1. We have shown that the mutagenic potency of these chemicals can be modulated by other dietary compounds, including green tea and conjugated linoleic acid, and the dioxin 2,3,7,8-tetrachlorodibenzo[b,e][1,4]dioxin (TCDD). These results demonstrate that the lacI transgenic rodent is a useful model for the study of chemoprevention in vivo. Publication Types: Review PMID: 11694652 [PubMed - indexed for MEDLINE] 1930: Nature. 2001 Nov 1;414(6859):1. Comment in: Nature. 2001 Dec 20-27;414(6866):843. Finding a future for GM crops. [No authors listed] Publication Types: Editorial PMID: 11689897 [PubMed - indexed for MEDLINE] 1931: Biotechnol Annu Rev. 2001;7:239-60. Transgenics in crops. Li Y, Wu YH, McAvoy R, Duan H. Collaborators: Li Y. Department of Plant Science, University of Connecticut, Storrs, CT 06269, USA. yi.li@uconn.edu With rapid world population growth and declining availability of fresh water and arable land, a new technology is urgently needed to enhance agricultural productivity. Recent discoveries in the field of crop transgenics clearly demonstrate the great potential of this technology for increasing food production and improving food quality while preserving the environment for future generations. In this review, we briefly discuss some of the recent achievements in crop improvement that have been made using gene transfer technology. Publication Types: Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Review PMID: 11686046 [PubMed - indexed for MEDLINE] 1932: New Sci. 1999 May 22;162(2187):3. The name of the game: the battle over genetically modified foods is not what it seems. [No authors listed] Publication Types: Editorial PMID: 11657971 [PubMed - indexed for MEDLINE] 1933: Smithsonian. 1998 Jul;29(4):22-30. New breeds down on the pharm: plain old barnyard animals -- with genes from other species added -- are producing medicines that keep people alive. Rosenfeld A. PMID: 11657703 [PubMed - indexed for MEDLINE] 1934: J Law Med Ethics. 1999 Summer;27(2):153-7. Commentary: a critique of Clark's frightening xenotransplantion scenario. Vanderpool HY. PMID: 11657463 [PubMed - indexed for MEDLINE] 1935: J Law Med Ethics. 1999 Summer;27(2):137-52. This little piggy went to market: the xenotransplantation and xenozoonose debate. Clark MA. PMID: 11657462 [PubMed - indexed for MEDLINE] 1936: J Biolaw Bus. 1998 Spring;1(3):3-7. Regulation of genetic modification of organisms: science or emotion? Malinowski MJ. PMID: 11657280 [PubMed - indexed for MEDLINE] 1937: Hastings Cent Rep. 1998 Sep-Oct;28(5):6. Power corrupts. Nicholson RH. PMID: 11656771 [PubMed - indexed for MEDLINE] 1938: NY Times (Print). 1998 Feb 3:F1, F6. Animals as organ donors? Not until they're germ-free. Stolberg SG. Publication Types: Newspaper Article PMID: 11647603 [PubMed - indexed for MEDLINE] 1939: Nat Biotechnol. 1998 Dec;16(13):1298. Europe's ethics standards. [No authors listed] PMID: 11645139 [PubMed - indexed for MEDLINE] 1940: Nature. 2001 Oct 18;413(6857):661. Bid to end EU's transgenic impasse. Schiermeier Q. PMID: 11606987 [PubMed - indexed for MEDLINE] 1941: Curr Opin Biotechnol. 2001 Oct;12(5):510-5. Edible genetically modified microorganisms and plants for improved health. Mercenier A, Wiedermann U, Breiteneder H. Laboratory of Bacteriology of Ecosystems, Institut Pasteur de Lille, 1 rue du Pr. Calmette, BP 245, F-59019 Cedex, Lille, France. annick.mercenier@ibl.fr The development of new strategies for the delivery of vaccine antigens or immune modulators to the mucosal tissue includes innovative approaches such as the use of genetically modified food microorganisms and plants. Even though the 'proof-of-concept' has recently been established for these two systems, key questions mainly related to efficacy and risk of breaking oral tolerance remain to be critically addressed in the immediate future. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 11604330 [PubMed - indexed for MEDLINE] 1942: Curr Opin Biotechnol. 2001 Oct;12(5):488-92. Plant-based raw material: improved food quality for better nutrition via plant genomics. van der Meer IM, Bovy AG, Bosch D. Plant Research International BV, Droevendaalsesteeg 1, PO Box 16, 6700AA, Wageningen, The Netherlands. Plants form the basis of the human food chain. Characteristics of plants are therefore crucial to the quantity and quality of human food. In this review, it is discussed how technological developments in the area of plant genomics and plant genetics help to mobilise the potential of plants to improve the quality of life of the rapidly growing world population. Publication Types: Review PMID: 11604326 [PubMed - indexed for MEDLINE] 1943: Biomed Environ Sci. 2001 Jun;14(1-2):66-74. Perspectives on nutrition needs for the new millennium for South Asian regions. Krishnaswamy K. National Institute of Nutrition, Indian Council of Medical Research, Hyderabad, AP, India. South Asia is the most populated region of the world with several nutritional challenges. Though per capita food energy supply, child survival and life expectancy have improved, and even today large segments of the population are below the poverty line with high infant and maternal mortality rates. It is important to recognize the crucial role of nutrition throughout the life cycle-from conception to old age. It is very necessary now to move from food security to nutrition security and improve the quality of foods both in macro- and micronutrients in order to break the transgenerational effects of malnutrition. The key solutions to the problems should address the issue of social development, population stabilization, environmental degradation and inadequate health and nutritional services. Strategies for empowering women and actuating community participation as sustainable programmes for human development, measures to reduce underweight and stunting in children and prevention of micronutrient malnutrition across the population are required. Enhancing food and nutrition security through innovative diversified agriculture and dietary practices, prevention and control of infection, promotion of food safety and fortification of staples with appropriate attention on emerging chronic disorders are essential. Population control measures to stabilize the fertility rates, biotechnological approaches for genetically modified foods, nutrition surveillance based on assessment, analysis and action to address the logistic, technical and compliance issues with emphasis on promotion of breast feeding and complementary foods with adequate attention on the reproductive needs of adolescent girls, pregnant mothers and lactating women would eliminate low birth weight, stunting, and chronic energy deficiency in vulnerable groups. Focused studies on bioavailability of micronutrients and its enhancement, innovative horticulture interventions, fortifications, social marketing strategies would promote the intake of micronutrient and phytonutrient rich foods. In-depth epidemiological research, an insight into foetal origins of adult disease and nutrition-genes interaction and life style alterations will avert the emerging epidemic of chronic diet related disorders. An investment in preventing foetal malnutrition improves nutrition of women in reproductive age, infant and child nutrition and prevents the onset of chronic disease in adult life. Human resource development, IEC measures, technology transfer, operational and logistic research, building of databases, integrated, intersectoral, multidisciplinary plans and sound management information system and surveillance with net working and experience sharing in the region will help to overcome the common challenges and lay the foundation for a better scenario in these regions in the near future. PMID: 11594482 [PubMed - indexed for MEDLINE] 1944: Biomed Environ Sci. 2001 Jun;14(1-2):40-3. Modern biotechnology-current standards in Japan. Nakamura Y. Standards Division, Department of Food Sanitation, Ministry of Health, Labour and Welfare, Tokyo, Japan. PMID: 11594478 [PubMed - indexed for MEDLINE] 1945: Biomed Environ Sci. 2001 Jun;14(1-2):14-20. Practical approaches to risk assessment. Brooke-Taylor S. Australia New Zealand Food Authority, Canberra, ACT. The importance of using risk assessment in developing food regulations is growing with the globalization of our food supply. The World Trade Organization has entrenched the principles of science-based risk assessment in the Agreement on Sanitary and Phytosanitary Measures. The relevant international organization for food standards, the Codex Alimentarius Commission, recognises risk analysis, and its component parts risk assessment, risk management and risk communication, as the basis for scientific decision-making. Risk assessment comprises two activities: hazard evaluation; and exposure estimation. A hazard may be chemical, microbiological or nutritional in origin. The practical application of risk assessment in Australia is illustrated in this presentation by four examples involving: (1) food additives, (2) microbiological safety of imported raw milk cheeses, (3) genetically modified foods and (4) imported food inspection. PMID: 11594473 [PubMed - indexed for MEDLINE] 1946: J Toxicol Environ Health A. 2001 Sep;64(1-2):1-210. Elements of precaution: recommendations for the regulation of food biotechnology in Canada. Royal Society of Canada. Publication Types: Consensus Development Conference Review PMID: 11592267 [PubMed - indexed for MEDLINE] 1947: J Allergy Clin Immunol. 2001 Oct;108(4):654. Comment on: J Allergy Clin Immunol. 2001 May;107(5):765-71. Allergenic risk from novel plants is very important and difficult to predict. Bucchini L. Publication Types: Comment Letter PMID: 11590400 [PubMed - indexed for MEDLINE] 1948: Biologicals. 2001 Jun;29(2):81-95. A quantitative analysis for the ADP-ribosylation activity of pertussis toxin: an enzymatic-HPLC coupled assay applicable to formulated whole cell and acellular pertussis vaccine products. Cyr T, Menzies AJ, Calver J, Whitehouse LW. Research Services Division, Bureau of Biologics and Radiopharmaceuticals, Biologics and Genetic Therapies Directorate, Health Products and Food Branch, Health Canada, Tunney's Pasture, Ottawa, K1A 0L2, Canada. terry_cyr@hc-sc.gc.ca The majority of the biological effects of pertussis toxin (PT) are the result of a toxin-catalyzed transfer of an adenosine diphosphate-ribose (ADP-ribose) moiety from NAD(+)to the alpha-subunits of a subset of signal-transducing guanine-nucleotide-binding proteins (G-proteins). This generally leads to an uncoupling of the modified G-protein from the corresponding receptor and the loss of effector regulation. This assay is based on the PT S1 subunit enzymatic transfer of ADP-ribose from NAD to the cysteine moiety of a fluorescent tagged synthetic peptide homologous to the 20 amino acid residue carboxyl-terminal sequence of the alpha-subunit of the G(i3)protein. The tagged peptide and the ADP-ribosylated product were characterized by HPLC/MS and MS/MS for structure confirmation. Quantitation of this characterized ADP-ribosylated fluorescently tagged peptide was by HPLC fluorescence using Standard Addition methodology. The assay was linear over a five hr incubation period at 20 degrees C at PT concentrations between 0.0625 and 4.0 microg/ml and the sensitivity of the assay could be increased several fold by increasing the incubation time to 24 h. Purified S1 subunit of PT exhibited 68.1+/-10.1% of the activity of the intact toxin on a molar basis, whereas the pertussis toxin B oligomer, the genetically engineered toxoid, (PT-9K/129G), and several of the other components of the Bordetella pertussis organism possessed little (<0.6%) or no detectable ribosylation activity. Commonly used pertussis vaccine reference materials, US PV Lot #11, BRP PV 66/303, and BRP PV 88/522, were assayed by this method against Bordetella pertussis Toxin Standard 90/518 and demonstrated to contain, respectively, 0.323+/-0.007, 0.682+/-0.045, and 0.757+/-0.006 microg PT/ml (Mean+/-SEM) or in terms of microg/vial: 3.63, 4.09 and 4.54, respectively. A survey of several multivalent pertussis vaccine products formulated with both whole cell as well as acellular components indicated that products possessed a wide range of ribosylation activities. The pertussis toxin S1 subunit catalyzed ADP- ribosylation of the FAC-Galpha(i3)C20 peptide substrate and its subsequent quantitation by HPLC was demonstrated to be a sensitive and quantitative method for measuring intrinsic pertussis toxin activity. This methodology not only has the potential to be an alternative physicochemical method to replace existing bioassay methodology, but has the added advantage of being a universal method applicable to the assay of pertussis toxin in both whole cell and acellular vaccines as well as bulk and final formulated vaccine products. Acceptance of this method by regulatory agencies and industry as a credible alternative to existing methods would, however, require validation in an international collaborative study against the widely accepted bioassay methods. Copyright 2001 The International Association for Biologicals. Publication Types: In Vitro PMID: 11580213 [PubMed - indexed for MEDLINE] 1949: Shokuhin Eiseigaku Zasshi. 2001 Jun;42(3):197-201. [A detection method for recombinant DNA from genetically modified maize CBH351] [Article in Japanese] Matsuoka T, Kuribara H, Suefuji S, Miura H, Kusakabe Y, Akiyama H, Goda Y, Isshiki K, Toyoda M, Hino A. National Food Research Institute, MAFF: 2-1-2, Kannondai, Tsukuba, Ibaraki 305-8642, Japan. A method using polymerase chain reaction (PCR) was designed for the detection of genetically modified maize CBH351, which has not authorized as safe for use in foods and feeds in Japan yet. We analyzed a recombinant DNA (r-DNA) sequence introduced into CBH351 maize and designed specific primer pairs to amplify a segment including part of the r-DNA. The PCR products obtained by using the designed primer pairs are specific for CBH351 and should prevent false positive results caused by other maizes and other main cereal crops. The r-DNA introduced into CBH351 could be detected from maize samples containing 0.05-0.1% CBH351 maize. This sensitivity is theoretically equivalent to a level of several genome copies and so this technique is a very efficient means to detect CBH351 maize. Publication Types: English Abstract Research Support, Non-U.S. Gov't PMID: 11577393 [PubMed - indexed for MEDLINE] 1950: Plant J. 2001 Sep;27(6):503-28. Assessment of the food safety issues related to genetically modified foods. Kuiper HA, Kleter GA, Noteborn HP, Kok EJ. National Institute for Quality Control of Agricultural Products (RIKILT), Wageningen University and Research Centre, PO Box 230, NL 6700 AE Wageningen, The Netherlands. h.a.kuiper@rikilt.wag-ur.nl International consensus has been reached on the principles regarding evaluation of the food safety of genetically modified plants. The concept of substantial equivalence has been developed as part of a safety evaluation framework, based on the idea that existing foods can serve as a basis for comparing the properties of genetically modified foods with the appropriate counterpart. Application of the concept is not a safety assessment per se, but helps to identify similarities and differences between the existing food and the new product, which are then subject to further toxicological investigation. Substantial equivalence is a starting point in the safety evaluation, rather than an endpoint of the assessment. Consensus on practical application of the principle should be further elaborated. Experiences with the safety testing of newly inserted proteins and of whole genetically modified foods are reviewed, and limitations of current test methodologies are discussed. The development and validation of new profiling methods such as DNA microarray technology, proteomics, and metabolomics for the identification and characterization of unintended effects, which may occur as a result of the genetic modification, is recommended. The assessment of the allergenicity of newly inserted proteins and of marker genes is discussed. An issue that will gain importance in the near future is that of post-marketing surveillance of the foods derived from genetically modified crops. It is concluded, among others that, that application of the principle of substantial equivalence has proven adequate, and that no alternative adequate safety assessment strategies are available. Publication Types: Review PMID: 11576435 [PubMed - indexed for MEDLINE] 1951: Toxicol Sci. 2001 Oct;63(2):153-6. Genetically modified plants and human health risks: Can additional research reduce uncertainties and increase public confidence? Hodgson E. Department of Environmental and Molecular Toxicology, Box 7633, North Carolina State University, Raleigh, North Carolina 27695, USA. ernest_hodgson@ncsu.edu So long as the risks to human health from transgenic plants remain potential rather than actual, and, in any event, appear lower than those from traditional plant breeding, hazard assessment need not be extensive. However, in view of current public attitudes to transgenic plants, it is necessary that those tests that are required, be based on logic, on sound science, and in accordance with the best scientific methodology. This is particularly the case with testing for food allergenicity. Current testing is largely indirect and based on comparisons with other known food allergens. Development of direct tests that involve interaction between the actual transgenic protein in question and the immune system is essential if confidence in the regulatory system is to be restored. Publication Types: Comparative Study PMID: 11568357 [PubMed - indexed for MEDLINE] 1952: EMBO Rep. 2001 Sep;2(9):744-5. Comment on: EMBO Rep. 2001 Aug;2(8):644-7. EMBO Rep. 2001 Jun;2(6):455-9. Comment on Stefan Flothmann and Jan van Aken's article 'Of maize and men' in EMBO reports, August 2001. Leaver CJ, Trewavas AJ. Publication Types: Comment Letter PMID: 11559579 [PubMed - indexed for MEDLINE] 1953: Plant Physiol. 2001 Sep;127(1):222-9. Overexpression of acyl carrier protein-1 alters fatty acid composition of leaf tissue in Arabidopsis. Branen JK, Chiou TJ, Engeseth NJ. University of Illinois, Department of Food Science and Human Nutrition, 259 ERML, 1201 West Gregory, Urbana, Illinois 61801, USA. Acyl carrier protein (ACP) is a small (9 kD) acidic protein that is an essential cofactor in plant fatty acid biosynthesis. Most plants have several isoforms of ACP, some of which are expressed constitutively and others that appear to be more tissue specific. Although the critical role of ACP in fatty acid biosynthesis has been established, the role of the diverse number of isoforms has yet to be elucidated. We have generated transgenic Arabidopsis plants that express high levels of ACP-1, a seed-predominant ACP isoform, in leaf tissue under control of the cauliflower mosaic virus 35S promoter. Western and northern analysis of these plants demonstrate 3- to 8-fold increased expression of this isoform in leaf tissue, but no significant changes in seed. Analysis of the fatty acid composition of leaf tissue revealed that overexpression of ACP-1 in leaf tissue alters fatty acid composition. Significant decreases in levels of 16:3 were noted along with increases in 18:3. These findings represent the first in vivo report that overexpression of an ACP isoform results in changes in fatty acid composition in plants. Publication Types: Research Support, Non-U.S. Gov't PMID: 11553750 [PubMed - indexed for MEDLINE] 1954: Virology. 2001 Sep 15;288(1):18-28. Trans-complementation of long-distance movement of White clover mosaic virus triple gene block (TGB) mutants: phloem-associated movement of TGBp1. Lough TJ, Emerson SJ, Lucas WJ, Forster RL. Plant Health and Development Group, Horticulture and Food Research Institute of New Zealand, Palmerston North, New Zealand. t.lough@genesis.co.nz The triple gene block proteins (TGBp1-3) and coat protein (CP) of potexviruses are required for cell-to-cell movement. Both cell-to-cell and long-distance movement of White clover mosaic virus in which individual, combinations, or all movement functions were mutated could be rescued by transgenic Nicotiana benthamiana expressing complementary viral products. To address the importance of TGB functions in vascular transport, we used an experimental system based on grafted plants and trans-complementation, to define co-translocated viral products and the minimal requirements for viral exit from the plant vasculature. Evidence is presented that TGBp1 is co-translocated with viral RNA and CP and that, once viral RNA is loaded into the phloem translocation stream, it can exit in sink tissues and replicate in the absence of TGBp2-3. These results are discussed in the context of the recent finding that TGBp1 can mediate the suppression of signaling involved in systemic gene silencing. Copyright 2001 Academic Press. Publication Types: Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. PMID: 11543654 [PubMed - indexed for MEDLINE] 1955: Nat Biotechnol. 2001 Sep;19(9):870-5. Comment in: Nat Biotechnol. 2001 Sep;19(9):826-7. Stable genetic transformation of tomato plastids and expression of a foreign protein in fruit. Ruf S, Hermann M, Berger IJ, Carrer H, Bock R. Institut für Biologie III, Universität Freiburg, Schänzlestrasse 1, D-79104 Freiburg, Germany. Transgenic chloroplasts offer unique advantages in plant biotechnology, including high-level foreign protein expression, absence of epigenetic effects, and gene containment due to the lack of transgene transmission through pollen. However, broad application of plastid genome engineering in biotechnology has been largely hampered by both the lack of chloroplast transformation systems for major crop plants and the usually low plastid gene expression levels in nongreen tissues such as fruits, tubers, and other storage organs. Here we describe the development of a plastid transformation system for tomato, Lycopersicon esculentum. This is the first report on the generation of fertile transplastomic plants in a food crop with an edible fruit. We show that chromoplasts in the tomato fruit express the transgene to approximately 50% of the expression levels in leaf chloroplasts. Given the generally very high foreign protein accumulation rates that can be achieved in transgenic chloroplasts (>40% of the total soluble protein), this system paves the way to efficient production of edible vaccines, pharmaceuticals, and antibodies in tomato. Publication Types: Research Support, Non-U.S. Gov't PMID: 11533648 [PubMed - indexed for MEDLINE] 1956: J Hazard Mater. 2001 Sep 14;86(1-3):205-22. Democracy and the governance of uncertainty. The case of agricultural gene technologies. Pellizzoni L. Department of Human Sciences, University of Trieste, Piazzale Europa 1, 34127, Trieste, Italy. pellizzonil@sp.univ.trieste.it The use of genetically modified organisms (GMOs) in agriculture and food production is the object of an intense and divisive debate. Drawing on a study on the public perception of agricultural gene technologies carried out in five European countries, the article deals with the policy aspects of the issue, and more precisely on the relation between institutions, experts and the public in a context of deep uncertainty. A theoretical framework is developed and compared with the study findings, suggesting that issues like the GMOs one represent a strong case for a more participatory policy-making. My conclusions suggest a style of governance based on the principles of deliberative democracy, as a suitable approach to the confrontation of different viewpoints and forms of knowledge. This appears to be the best way to improve the overall quality of policy-making: in this I include its legitimacy, the degree of public trust, and also the actual quality of its products. Strengthening the role of the public sphere seems more effective than simply increasing direct decision-making by the populace, and it offers an alternative to the 'elitist' solutions to the crisis of representative democracy. PMID: 11532367 [PubMed - indexed for MEDLINE] 1957: Aust N Z J Public Health. 2001 Aug;25(4):371-5. Health and safety issues pertaining to genetically modified foods. Goodyear-Smith F. Division of General Practice and Primary Health Care, Faculty of Medical and Health Sciences, University of Auckland, New Zealand. f.goodyear-smith@auckland.ac.nz Genetic modification involves the insertion of genes from other organisms (within or between species) into host cells to select for desirable qualities. Potential benefits of GM foods include increased nutritional value; reduced allergenicity; pest and disease-resistance; and enhanced processing value. Possible detrimental outcomes include producing foods with novel toxins, allergens or reduced nutritional value, and development of antibiotic resistance or herbicide-resistant weeds. Benefits to individuals or populations need to be weighed against adverse health and environmental risks, and may differ between developing and Westernised countries. Whether testing and monitoring should exceed requirements for conventional foods is under debate. While not necessarily scientifically justifiable, consumer concerns have resulted in Australian and New Zealand requirements to label foods containing GM-produced proteins. Dissatisfied consumer advocacy groups are calling for all foods involving GM technology to be labelled, irrelevant of whether the final product contains novel protein. Goals to improve the quantity, quality and safety of foods are laudable; however, the primary aim of the bio-food industry is financial gain. GM foods may be as safe as conventional foods but public distrust runs high. It is important that discussion is informed by science and that claims of both benefits and risks are evidence-based, to ensure that the process is driven neither by the vested interest of the bio-technical multinational companies on the one hand, nor ill-informed public fears on the other. PMID: 11529622 [PubMed - indexed for MEDLINE] 1958: J Environ Monit. 1999 Dec;1(6):108N-110N. Genetically modified organisms and monitoring. Diamand E. The genetic modification of organisms for food use has raised serious concern about the potential for adverse effects on the environment, ecosystems and on the health of humans and animals. As a relatively new technology, its impacts remain uncertain but could range from disturbances to the genetic functioning of individual organisms to a reduction in the biodiversity of farmland. As a result, the question of how to monitor for potential impacts is beset with problems. The fact that genetic modification can be used on a range of organisms for a variety of purposes means that those developing monitoring systems will need to be as imaginative as those developing GMOs. In the case of genetically modified organisms (GMOs) for food use, concern has focussed on the transfer of genes to other organisms, the potential for effects on non-target organisms, or on the health of humans and animals, and the likelihood of adverse effects on wildlife due to changes in farming practice. As with other new and unfamiliar technologies, genetic modification is also plagued by the problem of uncertainty. Novel genes are inserted randomly into the genome of the host organisms, and this leads to the possibility of unexpected effects. Unanticipated environmental disasters, such as the concentration of persistent organic pollutants in ecosystems at high latitudes, have highlighted the need for monitoring despite the obvious difficulties inherent in monitoring for unexpected effects. PMID: 11529177 [PubMed - indexed for MEDLINE] 1959: Trends Biotechnol. 2001 Sep;19(9):333. Is seed-contamination with GMOs a problem for food safety and the environment? Halsberger AG. Institute of Microbiology and Genetics, University of Vienna, Austria. hasi1@via.at Publication Types: Letter PMID: 11525200 [PubMed - indexed for MEDLINE] 1960: Mol Genet Genomics. 2001 Jul;265(5):763-70. Enhancers and core promoter elements are essential for the activity of a cryptic gene activation sequence from tobacco, tCUP. Wu K, Malik K, Tian L, Hu M, Martin T, Foster E, Brown D, Miki B. Eastern Cereal and Oilseed Research Centre, Agriculture and Agri-Food Canada, Ottawa, Ontario. Cryptic gene regulatory elements are sequences that are inactive at their native locations in the genome but have the ability to become functional when positioned adjacent to genes. We have recently isolated such a cryptic sequence from tobacco, tCUP, that can act as a promoter. A 135-bp fragment spanning extending from position -197 to -62, relative to the transcription start site, was found to promote GUS expression in all of the major organs of transgenic Arabidopsis plants. Furthermore, this 135-bp fragment complemented the -46 minimal promoter of CaMV 35S and conferred constitutive expression on transgenic Arabidopsis plants. An electrophoretic mobility-shift assay showed that nuclear proteins prepared from tobacco leaves interact with the 135-bp fragment. tCUP has a core promoter that lacks the TATA consensus sequence but addition of a TATA-box sequence increased the core promoter activity by three-fold. The sequence surrounding the transcription start site of tCUP has sequence similarity with the initiator element (Inr), and deletion of this sequence significantly reduced promoter activity, suggesting that an essential Inr element may exist in the tCUP core promoter. Fusion of the GCC-box enhancer element from pathogenesis-related genes to the core promoter elevated tCUP core promoter activity. Our study indicates that cryptic promoters are similar in composition and organization to promoters associated with expressed genes and that their promoter elements can be combined to create composite promoters that are fully functional. This data provides direct evidence that the expression pattern of plant genes can be influenced by cryptic gene regulatory elements when they are brought into juxtaposition with genes through DNA rearrangements. Publication Types: Research Support, Non-U.S. Gov't PMID: 11523793 [PubMed - indexed for MEDLINE] 1961: Proc Natl Acad Sci U S A. 2001 Sep 11;98(19):10775-80. Epub 2001 Aug 21. Tetracycline-inducible systems for Drosophila. Stebbins MJ, Urlinger S, Byrne G, Bello B, Hillen W, Yin JC. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY 11724, USA. Since their inception, tetracycline (Tet)-inducible systems have become the method of choice for transgenic research. The Tet-Off systems have a number of advantages, including robust target induction using a relatively benign effector molecule. However, use of the Tet-On system has been fraught with difficulties, including high background expression in the absence of effector molecules and inconsistent gene induction. Recently, second generation Tet-On transactivators (TAs) have been described. In HeLa cells, they are far more efficient than the original reverse TA protein, and they exhibit lower background activity in the absence of effectors. Here we examine the most promising TA in transgenic Drosophila and characterize its in vivo properties. We report that low levels of doxycycline, when added to normal fly food, efficiently and rapidly induce target transgenes in adults, larvae, and embryos. This TA is superior to all other Tet-On proteins, and its performance is comparable to that of the widely used Tet-Off TA. In addition, combining the improved Tet-On TA with the Gal4-UAS (upstream-activating sequence) system produces robust, spatially restricted, temporally controlled transgene induction. Because this Tet-On TA is significantly more efficient than previous ones used in Drosophila, it is also possible to modulate gene induction by controlling the dosage of the antibiotic in the food. Publication Types: Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. PMID: 11517299 [PubMed - indexed for MEDLINE] 1962: Plant Mol Biol. 2001 Jul;46(5):531-8. Identification of pronp1, a tobacco profilin gene activated in tip-growing cells. Swoboda I, Bhalla PL, Xu H, Zhang Y, Mittermann I, Valenta R, Singh MB. Plant Molecular Biology and Biotechnology Laboratory, Institute of Land & Food Resources, University of Melbourne, Parkville, Vic., Australia. In plant cells, several cellular processes depend on rapid reorganization of a dynamic network of actin cytoskeletal elements in response to internal and environmental stimuli. Profilins, ubiqitous eukaryotic actin monomer-binding proteins with highly conserved three-dimensional structures, regulate the actin cytoskeleton and are considered to link the microfilament system with signal transduction pathways. Plant profilins have been grouped into two distinct classes, gametophytic (pollen-specific) and sporophytic. Here we report the isolation of a profilin gene that seems to be activated during tip growth of specialized cells of gametophytic as well as sporophytic origin. Identification of a genomic DNA clone containing a tobacco profilin gene, pronp1, and analysis of the pronp1 promoter-uidA fusion gene in transgenic Nicotiana tabacum plants revealed a prominent expression of pronp1 in mature pollen and elongating pollen tubes and significant activity in root hairs of developing seedlings. This expression pattern was distinct from that of any other profilin gene isolated so far. Pronp1 thus represents a unique profilin gene that is activated at the transcriptional level in two kinds of tip-growing cells, pollen tubes and root hairs, both of which require rapid organization of the actin cytoskeleton. The isolation of such a gene has fundamental importance for our understanding of modulation of the actin cytoskeleton at the molecular level. Publication Types: Research Support, Non-U.S. Gov't PMID: 11516146 [PubMed - indexed for MEDLINE] 1963: Biotechniques. 2001 Aug;31(2):426-9. PCR-based detection of genetically modified soybean and maize in raw and highly processed foodstuffs. Tengel C, Schüssler P, Setzke E, Balles J, Sprenger-Haussels M. QIAGEN GmbH, Hilden, Germany. The PCR method has proved to be an invaluable tool for the specific and sensitive detection of genetically modified material (e.g., Roundup Ready Soybean and Bt-176 "Maximizer" Maize) in foodstuffs. The first step in the procedure, namely the purification of nucleic acids from the sample, is often the deciding factor in the production of meaningful results. In this study, we present two procedures that enable an efficient isolation of trace amounts of genetic material from both raw and highly processed foodstuffs. We show that for optimal, PCR-ready DNA purification from highly processed foodstuffs and PCR inhibitor-rich substances--such as cocoa-containing products--adapted protocols for the QIAGEN QIAamp DNA Stool Mini Kit can be utilized. For complete DNA isolation from raw foodstuffs, a protocol using the DNeasy Plant Mini Kit is presented. PMID: 11515380 [PubMed - indexed for MEDLINE] 1964: J Agric Food Chem. 2001 Aug;49(8):3638-43. Increase in lipid content in potato tubers modified by 14-3-3 gene overexpression. Prescha A, Swiedrych A, Biernat J, Szopa J. Department of Food Science and Nutrition, Medical University, Nankiera 1, 50-140 Wrocław, Poland. Recently, transgenic potato plants were created with overexpression of the 14-3-3 protein derived from Cucurbita pepo. Detailed analysis of those plants suggested that the function of the isolated 14-3-3 isoform is in the control of carbohydrate and lipid metabolism in the plants. 14-3-3 protein overexpression gave rise to an increase in soluble sugar and catecholamine contents in both leaves and tubers. It is proposed that 14-3-3 protein affects carbohydrate metabolism in potato plants via regulation of catecholamine synthesis. Furthermore, genetically modified potato tubers with 14-3-3 protein overexpression showed changes in lipid content and composition. The transgenic potato tubers contained 69% more total fat compared to the wild-type plant. Separation of tuber lipids into polar and nonpolar fractions revealed that the transgenic potato tubers contained almost 3 times more nonpolar lipids than the control. Analysis of fatty acid composition, conducted by the means of gas chromatography, showed that linoleic acid was the main fatty acid present in the tubers of both modified and control potato plants. In the nonpolar fraction of the fat of the transgenic tubers the unsaturated fatty acids exhibited a higher participation in the sum of all fatty acids. Publication Types: Research Support, Non-U.S. Gov't PMID: 11513640 [PubMed - indexed for MEDLINE] 1965: J Agric Food Chem. 2001 Aug;49(8):3622-7. A rapeseed-specific gene, acetyl-CoA carboxylase, can be used as a reference for qualitative and real-time quantitative PCR detection of transgenes from mixed food samples. Hernández M, Río A, Esteve T, Prat S, Pla M. Instituto de Biología Molecular de Barcelona-Consejo Superior de Investigaciones Científicas, Jordi Girona 18-26, 08034 Barcelona, Spain. Polymerase chain reaction (PCR) methods are very useful techniques for the detection and quantification of genetically modified organisms (GMOs) in food samples. These methods rely on the amplification of transgenic sequences and quantification of the transgenic DNA by comparison to an amplified reference gene. Reported here is the development of specific primers for the rapeseed (Brassica napus) BnACCg8 gene and PCR cycling conditions suitable for the use of this sequence as an endogenous reference gene in both qualitative and quantitative PCR assays. Both methods were assayed with 20 different rapeseed varieties, and identical amplification products were obtained with all of them. No amplification products were observed when DNA samples from other Brassica species, Arabidopsis thaliana, maize, and soybean were used as templates, which demonstrates that this system is specific for rapeseed. In real-time quantitative PCR analysis, the detection limit was as low as 1.25 pg of DNA, which indicates that this method is suitable for use in processed food samples which contain very low copies of target DNA. Publication Types: Research Support, Non-U.S. Gov't PMID: 11513638 [PubMed - indexed for MEDLINE] 1966: Ital J Biochem. 2000 Sep-Dec;49(3-4):64-72. Biosensors as a new analytical tool for detection of genetically modified organisms. Minunni M, Mascini M, Motti C, Dainese E, Di Matteo A, Chen B, Cozzani I. University of Teramo, Faculty Agriculture and Food Science Mosciano Stazione, TE, 64023, Italy. PMID: 11508061 [PubMed - indexed for MEDLINE] 1967: Ital J Biochem. 2000 Sep-Dec;49(3-4):61-3. Quantitative analysis of GMO food contaminations using real time PCR. Locatelli G, Urso V, Malnati M. Pharmacia & Upjohn, Via Pasteur 10, 20014 Nerviano, MI. PMID: 11508060 [PubMed - indexed for MEDLINE] 1968: Ital J Biochem. 2000 Sep-Dec;49(3-4):56-61. Support to the implementation of European Commission policy in the area of GMOs. Querci M, van den Eede G, Eyquem F, Anklam E. PMID: 11508059 [PubMed - indexed for MEDLINE] 1969: Ital J Biochem. 2000 Sep-Dec;49(3-4):52-6. The use of biotechnology in agriculture and the methods for the detection of genetically modified organisms (GMOs) in food. Motti C, Dainese E, Mascini M, Minunni M, De Santis P, Cozzani I. Department of Structure, Function and Pathology of Animals and Biotechnology, University of Teramo, Italy. PMID: 11508058 [PubMed - indexed for MEDLINE] 1970: Mutagenesis. 2001 Sep;16(5):431-7. Mutations induced by 2-amino-1-methyl-6-phenylimidazo [4,5-b]pyridine (PhIP) in cecum and proximal and distal colon of lacI transgenic rats. Stuart GR, de Boer JG, Haesevoets R, Holcroft J, Kangas J, Sojonky K, Thorleifson E, Thornton A, Walsh DF, Yang H, Glickman BW. Centre for Environmental Health and Department of Biology, University of Victoria, PO Box 3020 STN CSC, Victoria, British Columbia, Canada V8W 3N5. stuart@niehs.nih.gov 2-Amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) is a food-borne mutagen and carcinogen that induces tumors of the colon and the prostate gland in male rats and of the mammary gland in female rats. In this study we describe the frequency and specificity of PhIP-induced mutations in the cecum, proximal colon and distal colon of male and female lacI transgenic rats. This is the first report of mutational data from discrete regions of the colon. After 61 days of treatment with 200 p.p.m. PhIP mixed into the diet, PhIP-induced mutant frequencies were elevated 7-fold in the cecum and 14- to 21-fold in the colon of male and female rats compared with untreated controls. PhIP-induced mutant frequencies increased significantly (overall trend, P < 10(-4)) along the length of the colon of both males and females, with cecum < proximal colon < distal colon. A total of 754 PhIP mutants (363 male, 391 female) were sequenced to provide the mutational spectra for each of the three tissue sections from males and females. These mutational spectra consisted predominantly of G:C-->T:A and G:C-->C:G transversions and deletions of G:C base pairs. There were no significant differences between the mutational spectra with respect to sex or position in the colon. Therefore, we surmise that following induction of mutations by PhIP in male and female colons, non-mutagenic factors, possibly hormonal, preferentially influence the formation of tumors in the colon of male rats. PMID: 11507243 [PubMed - indexed for MEDLINE] 1971: Br J Nutr. 2001 Aug;86(2):123-39. Uncoupling proteins: their roles in adaptive thermogenesis and substrate metabolism reconsidered. Dulloo AG, Samec S. Institute of Physiology, Department of Medicine, University of Fribourg, Rue du Musée 5, Fribourg, Switzerland. abdul.dulloo@unifr.ch During the past few years, there have been two major developments, if not revolutions, in the field of energy balance and weight regulation. The first at the molecular level, which was catalysed by developments in DNA screening technology together with the mapping of the human genome, has been the tremendous advances made in the identification of molecules that play a role in the control of food intake and metabolic rate. The second, at the systemic level, which centered upon the use of modern technologies or more robust analytical techniques for assessing human energy expenditure in response to starvation and overfeeding, has been the publication of several papers providing strong evidence that adaptive thermogenesis plays a much more important role in the regulation of body weight and body composition than previously thought. Within these same few years, several new members of the mitochondrial carrier protein family have been identified in a variety of tissues and organs. All apparently possess uncoupling properties in genetically-modified systems, with two of them (uncoupling protein (UCP) 2 and UCP3) being expressed in adipose tissues and skeletal muscles, which are generally recognised as important sites for variations in thermogenesis and/or in substrate oxidation. Considered as breakthrough discoveries, the cloning of these genes has generated considerable optimism for rapid advances in our molecular understanding of adaptive thermogenesis, and for the identification of new targets for pharmacological management of obesity and cachexia. The present paper traces first, from a historical perspective, the landmark events in the field of thermogenesis that led to the identification of these genes encoding candidate UCP, and then addresses the controversies and on-going debate about their physiological importance in adaptive thermogenesis, in lipid oxidation or in oxidative stress. The general conclusion is that UCP2 and UCP3 may have distinct primary functions, with UCP3 implicated in regulating the flux of lipid substrates across the mitochondria and UCP2 in the control of mitochondrial generation of reactive oxygen species. The distinct functions of these two UCP1 homologues have been incorporated in a conceptual model to illustrate how UCP2 and UCP3 may act in concert in the overall regulation of lipid oxidation concomitant to the prevention of lipid-induced oxidative damage. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 11502224 [PubMed - indexed for MEDLINE] 1972: Mol Plant Microbe Interact. 2001 Aug;14(8):939-46. Dark green islands in plant virus infection are the result of posttranscriptional gene silencing. Moore CJ, Sutherland PW, Forster RL, Gardner RC, MacDiarmid RM. Horticulture and Food Research Institute of New Zealand Limited, Auckland. Dark green islands (DGIs) are a common symptom of plants systemically infected with a mosaic virus. DGIs are clusters of green leaf cells that are free of virus but surrounded by yellow, virus-infected tissue. We report here on two lines of evidence showing that DGIs are caused by posttranscriptional gene silencing (PTGS). First, transcripts of a transgene derived from the coat protein of Tamarillo mosaic potyvirus (TaMV) were reduced in DGIs relative to adjacent yellow tissues when the plants were infected with TaMV. Second, nontransgenic plants coinfected with TaMV and a heterologous virus vector carrying TaMV sequences showed reduced titers of the vector in DGIs compared with surrounding tissues. DGIs also were compared with recovered tissue at the top of transgenic plants because recovery has been shown previously to involve PTGS. Cytological analysis of the cells at the junction between recovered and infected tissue was undertaken. The interface between recovered and infected cells had very similar features to that surrounding DGIs. We conclude that DGIs and recovery are related phenomena, differing in their ability to amplify or transport the silencing signal. Publication Types: Research Support, Non-U.S. Gov't PMID: 11497465 [PubMed - indexed for MEDLINE] 1973: Vopr Pitan. 2001;70(2):3-7. [Hygiene and standards aspects of registration, marking and labeling of food products prepared from genetically modified sources] [Article in Russian] Onishchenko GG. The basic requirements to problems of registration, marking and labelling of the foodstuff prepared from generically modified sources are stated. The comparative analysis of the above-named problems in the different countries (USA, countries of EU, Russia etc.) is given. Publication Types: Comparative Study English Abstract PMID: 11494669 [PubMed - indexed for MEDLINE] 1974: EMBO Rep. 2001 Aug;2(8):644-7. Comment in: EMBO Rep. 2001 Sep;2(9):744-5. Comment on: EMBO Rep. 2001 Jun;2(6):455-9. Of maize and men. Is the endorsement of GM crops science or politics? Flothmann S, van Aken J. Greenpeace, Germany. stefan.flothmann@greenpeace.de Publication Types: Comment PMID: 11493587 [PubMed - indexed for MEDLINE] 1975: Shokuhin Eiseigaku Zasshi. 2001 Apr;42(2):J143-6. [Unique identification system for the safety assessment of genetically modified organisms; unit and transparency] [Article in Japanese] Maekawa T. PMID: 11486391 [PubMed - indexed for MEDLINE] 1976: Science. 2001 Sep 7;293(5536):1826-8. Epub 2001 Jul 26. Resistance to an herbivore through engineered cyanogenic glucoside synthesis. Tattersall DB, Bak S, Jones PR, Olsen CE, Nielsen JK, Hansen ML, Høj PB, Møller BL. Plant Biochemistry Laboratory, Department of Plant Biology, Centre for Molecular Plant Physiology, Department of Chemistry, Royal Veterinary and Agricultural University, 40 Thorvaldsensvej, DK-1871, Frederiksberg C, Denmark. The entire pathway for synthesis of the tyrosine-derived cyanogenic glucoside dhurrin has been transferred from Sorghum bicolor to Arabidopsis thaliana. Here, we document that genetically engineered plants are able to synthesize and store large amounts of new natural products. The presence of dhurrin in the transgenic A. thaliana plants confers resistance to the flea beetle Phyllotreta nemorum, which is a natural pest of other members of the crucifer group, demonstrating the potential utility of cyanogenic glucosides in plant defense. Publication Types: Research Support, Non-U.S. Gov't PMID: 11474068 [PubMed - indexed for MEDLINE] 1977: Appl Environ Microbiol. 2001 Aug;67(8):3434-9. Location effects of a reporter gene on expression levels and on native protein synthesis in Lactococcus lactis and Saccharomyces cerevisiae. Thompson A, Gasson MJ. Institute of Food Research, Colney, Norwich NR4 7UA, United Kingdom. arthur.thompson@bbsrc.ac.uk The engineering of industrially important genetically modified organisms by the integration of heterologous genes into the chromosome is often the method of choice for several reasons concerned with long-term stability, homogeneous population distribution, and the enabling of selection without the addition of antibiotics. However, integration may disrupt endogenous gene expression, giving rise to increased levels of toxic metabolic byproducts or activating otherwise silent genes. The position of integration of a foreign gene in the chromosome can also influence its expression levels, and this effect will be of relevance in terms of optimizing protein production parameters. In this study, we determine how the random integration of a foreign reporter gene might affect expression levels and assess the use of proteome analysis to investigate possible effects on synthesis of endogenous proteins in two important food-relevant microorganisms, Saccharomyces cerevisiae and Lactococcus lactis. Eleven L. lactis integrants carrying the gusA gene were analyzed, and expression levels were found to vary by a factor of threefold in contrast to expression levels of lacZ in 18 S. cerevisiae integrants, which showed a 14-fold variation. Of relevance to industry is whether any changes in expression levels might occur as a consequence of storage of the modified strains. Here it is also shown that the above differences in expression levels were not significantly affected by storage of frozen cultures over a period of several months. Analysis of the protein composition of the yeast and lactococcal integrant strains by separation on one-dimensional (1D) and 2D gels showed no significant variations in position beyond those observed in control samples. Publication Types: Research Support, Non-U.S. Gov't PMID: 11472915 [PubMed - indexed for MEDLINE] 1978: Nature. 2001 Jul 19;412(6844):257-8. Britain seeks transgenics deal to fend off transatlantic trade war. Dickson D. Publication Types: Congresses News PMID: 11460116 [PubMed - indexed for MEDLINE] 1979: Minerva Pediatr. 2001 Jun;53(3):199-210. Transgenic foods, pesticides, dioxin, passive smoke. Consequences on breast milk. [Article in English, Italian] Cantani A, Micera M. Division of Allergy and Clinical Immunology, Pediatric Department, University La Sapienza, Rome, Italy. acantani@pelagus.it In recent years the efforts of companies manufacturing cow milk (CM) formulas have led to the development and availability of special formulas, which have dramatically reduced the morbidity of infants with food allergy (FA). However, the safety of several infants and children with food allergy is put to a severe test by transgenic foods, pesticides, and dioxin entering the scenario, also provoking breast milk contamination. Regarding transgenic foods, it is significant the attitude taken against people attempting to call them Frankenstein food, whereas pesticides and dioxin present in dietary foods for infants and young children, after a first arising of alarmed and inflamed controversies, have almost fallen into oblivion. Several of these substances are able to trigger immune alterations. Recent reports have shown that pears can contain pesticides in 54% of cases, a finding which obliges us to review elimination diets devised for allergic babies. However, these foods are far from being ideal both from the nutritional adequacy and hypoallergenicity; moreover, passive smoke is now a genetic factor. We would like to stress, according to the Latin wisdom that stands on the portal of our Clinic in puero homo, which means. In infant is the seed of the future man, that our goal is not only to reduce morbidity and mortality, but mainly to insure the best quality of life both to infants and adults. Publication Types: Review PMID: 11455307 [PubMed - indexed for MEDLINE] 1980: Vopr Pitan. 2000;69(6):37-40. [Organization of State sanitary inspection of food products from genetically modified sources] [Article in Russian] Petukhov AI. Publication Types: Comparative Study PMID: 11452373 [PubMed - indexed for MEDLINE] 1981: Fresenius J Anal Chem. 2001 Jun;370(2-3):142-6. Recent production of candidate reference materials at IRMM. Kramer GN, Pauwels J, Le Guern L, Schimmel H, Trapmann S. European Commission, Joint Research Centre, Institute for Reference Materials and Measurements, Geel, Belgium. kramer@jrc.irmm.be In the execution of its mission to promote a common European measurement system in support of EU policies, IRMM's Reference Materials Unit is currently involved in preparation of proficiency-testing samples and candidate reference materials. Recent work related to bovine spongiform encephalopathy in cows, genetically modified organisms, and a variety of environmental materials is described. Publication Types: Review PMID: 11451225 [PubMed - indexed for MEDLINE] 1982: Plant Cell. 2001 Jul;13(7):1669-82. Rerouting the plant phenylpropanoid pathway by expression of a novel bacterial enoyl-CoA hydratase/lyase enzyme function. Mayer MJ, Narbad A, Parr AJ, Parker ML, Walton NJ, Mellon FA, Michael AJ. Division of Food Safety Science, Institute of Food Research, Norwich Research Park, Colney, Norwich NR4 7UA, United Kingdom. The gene for a bacterial enoyl-CoA hydratase (crotonase) homolog (HCHL) previously shown to convert 4-coumaroyl-CoA, caffeoyl-CoA, and feruloyl-CoA to the corresponding hydroxybenzaldehydes in vitro provided an opportunity to subvert the plant phenylpropanoid pathway and channel carbon flux through 4-hydroxybenzaldehyde and the important flavor compound 4-hydroxy-3-methoxybenzaldehyde (vanillin). Expression of the Pseudomonas fluorescens AN103 HCHL gene in two generations of tobacco plants caused the development of phenotypic abnormalities, including stunting, interveinal chlorosis and senescence, curled leaf margins, low pollen production, and male sterility. In second generation progeny, the phenotype segregated with the transgene and transgenic siblings exhibited orange/red coloration of the vascular ring, distorted cells in the xylem and phloem bundles, and lignin modification/reduction. There was depletion of the principal phenolics concomitant with massive accumulation of novel metabolites, including the glucosides and glucose esters of 4-hydroxybenzoic acid and vanillic acid and the glucosides of 4-hydroxybenzyl alcohol and vanillyl alcohol. HCHL plants exhibited increased accumulation of transcripts for phenylalanine ammonia-lyase, cinnamate-4-hydroxylase, and 4-coumarate:CoA ligase, whereas beta-1,3-glucanase was suppressed. This study, exploiting the ability of a bacterial gene to divert plant secondary metabolism, provides insight into how plants modify inappropriately accumulated metabolites and reveals the consequences of depleting the major phenolic pools. Publication Types: Research Support, Non-U.S. Gov't PMID: 11449058 [PubMed - indexed for MEDLINE] 1983: FDA Consum. 2001 Mar-Apr;35(2):9-11. Proposed rules issued for bioengineered foods. Formanek R Jr. PMID: 11444252 [PubMed - indexed for MEDLINE] 1984: Arch Pharm Res. 2001 Jun;24(3):256-61. Allergenicity test of genetically modified soybean in Sprague Dawley rats. Chang HS, Bae YK, Lim SK, Jeong TC, Kim HS, Chung ST, Kim DS, Nam DH. College of Pharmacy, Yeungnam University, Kyongsan, Korea. Allergenicity of genetically-modified (GM) soybean was evaluated in male Sprague Dawley rats. To confirm the GM soybean used in this study, the polymerase chain reaction (PCR) was performed using the chromosomal DNA of soybeans. The PCR result provided the clear discrimination of genetically-modified (GM) soybeans. To evaluate the allergenicity of GM soybean and non-GM control one, the soybean homogenate was sensitized subcutaneously 3 times a week for 3 weeks. The doses of soybean were 0, 2 and 20 mg/kg in the protein basis. A week after the last sensitization, antisera were recovered from individual animals. When the sera were injected intradermally on the clipped back of unsensitized rats with various dilutions, followed by a challenge with 20 mg/kg of soybean homogenate containing 1% Evans blue, no sign of passive cutaneous anaphylaxis reaction was detected. In addition, when the sera were treated in the cultures of peritoneal mast cells, the increase of histamine release by anti-(GM soybean) sera was not observed when compared to that by anti-(non-GM soybean) sera. The present results indicate that the GM soybean might not act as a strong allergen in male Sprague Dawley rats. Publication Types: Research Support, Non-U.S. Gov't PMID: 11440087 [PubMed - indexed for MEDLINE] 1985: Plant Mol Biol. 2001 May;46(1):109-20. Activation of tomato PR and wound-related genes by a mutagenized tomato MAP kinase kinase through divergent pathways. Xing T, Malik K, Martin T, Miki BL. Agriculture and Agri-Food Canada, Eastern Cereal and Oilseed Research Centre, Ottawa, Ontario. A mitogen-activated protein kinase kinase (MAPKK) gene, tMEK2, was isolated from tomato cv. Bonny Best. By mutagenesis, a permanently active variant, tMEK2MUT, was created. Both wild-type tMEK2 and mutant tMEK2MUT were driven by a newly described strong plant constitutive promoter, tCUP, in a tomato protoplast transient gene expression system. Pathogenesis-related genes, PRlb1, PR3 and Twi1, and a wound-inducible gene, ER5, were activated by tMEK2MUT. Specific inhibitors of p38 class MAPK inhibited tMEK2MUT-induced activation of PR3 and ER5 genes but not that of the PRlb1 or Twi1 gene. Arabidopsis dual-specificity protein tyrosine phosphatase 1 (DsPTP1) and maize protein phosphatase 1 (PP1) inhibited tMEK2MUT-induced activation of the ER5 gene and the Twi1 gene, respectively, whereas PRlb1 and PR3 were not affected by either AtDsPTP1, or maize PP1, or Arabidopsis protein phosphatase 2A (PP2A). We have demonstrated for the first time that a single MAPKK activates an array of PR and wound-related genes. Our observation indicates that the activation of the genes downstream of tMEK2 occurs through divergent pathways and that tMEK2 may play an important role in the interaction of signal transduction pathways that mediate responses to both biotic (e.g. disease) and abiotic stresses (e.g. wound responsiveness). Publication Types: Research Support, Non-U.S. Gov't PMID: 11437246 [PubMed - indexed for MEDLINE] 1986: Mar Pollut Bull. 2001 May;42(5):335. Opposition to release of G.M. salmon. [No authors listed] Publication Types: News PMID: 11436812 [PubMed - indexed for MEDLINE] 1987: J Chromatogr B Biomed Sci Appl. 2001 May 25;756(1-2):327-35. Genetic modification and plant food allergens: risks and benefits. Shewry PR, Tatham AS, Halford NG. Department of Agricultural Sciences, University of Bristol, UK. peter.shewry@bbsrc.ac.uk Plant genetic engineering has the potential to both introduce new allergenic proteins into foods and remove established allergens. A number of allergenic plant proteins have been characterized, showing that many are related to proteins which have potentially valuable properties for use in nutritional enhancement, food processing and crop protection. It is therefore important to monitor the allergenic potential of proteins used for plant genetic engineering and major biotechnology companies have established systems for this. Current technology allows gene expression to be down-regulated using antisense or co-suppression and future developments may allow targeted gene mutation or gene replacement. However, the application of this technology may be limited at least in the short term by the presence of multiple allergens and their contribution to food processing or other properties. Furthermore, the long-term stability of these systems needs to be established as reversion could have serious consequences. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 11419724 [PubMed - indexed for MEDLINE] 1988: EMBO Rep. 2001 Jun;2(6):455-9. Comment in: EMBO Rep. 2001 Aug;2(8):644-7. EMBO Rep. 2001 Sep;2(9):744-5. Is opposition to GM crops science or politics? An investigation into the arguments that GM crops pose a particular threat to the environment. Trewavas A, Leaver C. Institute of Cell and Molecular Biology at the University of Edinburgh. trewavas@srv0.bio.ed.ac.uk PMID: 11415971 [PubMed - indexed for MEDLINE] 1989: Curr Opin Biotechnol. 2001 Jun;12(3):308-11. The regulation of biologic products derived from bioengineered plants. Stein KE, Webber KO. Division of Monoclonal Antibodies, Office of Therapeutics Research and Review, Center for Biologics Evaluation and Research, 29 Lincoln Drive, 20892-4555, Bethesda, MD, USA. stein@cber.fda.gov Recently, there has been a large increase in the number and types of biological products--from therapeutic antibodies to vaccines for the prevention of infectious diseases--that are produced in bioengineered plant systems. We anticipate that this technology will be used increasingly on a commercial scale for the manufacture of human and animal products. These production systems have the capacity to produce very large quantities of products at lower costs and with reduced risks compared with mammalian systems. Publication Types: Review PMID: 11404111 [PubMed - indexed for MEDLINE] 1990: Gene. 2001 May 30;270(1-2):103-11. Adaptable doxycycline-regulated gene expression systems for Drosophila. Stebbins MJ, Yin JC. Cold Spring Harbor Laboratory, 1 Bungtown Road, Cold Spring Harbor, NY 11724, USA. We have engineered two new versions of the doxycycline (dox) inducible system for use in Drosophila. In the first system, we have used the ubiquitously expressed Drosophila actin5C promoter to express the Tet-Off transactivator (tTA) in all tissue. Induction of a luciferase target transgene begins 6 h after placing the flies on dox-free food. Feeding drug-free food to mothers results in universal target gene expression in their embryos. Larvae raised on regular food also show robust expression of a target reporter gene. In the second version, we have used the Gal4-UAS system to spatially limit expression of the transactivator. Dox withdrawal results in temporally- and spatially-restricted, inducible expression of luciferase in the adult head and embryo. Both the actin5C and Gal4-UAS versions produce more than 100-fold induction of luciferase in the adult, with virtually no leaky expression in the presence of drug. Reporter gene expression is also undetectable in larvae or embryos from mothers fed dox-containing food. Such tight control may be due to the incorporation of Drosophila insulator elements (SCS and SCS') into the transgenic vectors. These systems offer a practical, effective alternative to currently available expression systems in the Drosophila research community. Publication Types: Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. PMID: 11404007 [PubMed - indexed for MEDLINE] 1991: Med Law. 2001;20(1):133-41. Ethical acceptability, health policy and foods biotechnology based foods: is there a third way between the precaution principle and an overly enthusiastic dissemination of GMO? Meningaud JP, Moutel G, Hervé C. Department of Medical Ethics and Public Health, Necker University Hospital, Paris, France. The demand for consumer safety with regard to the food-processing industry is becoming, legitimately, more and more urgent. If ingested drugs can carry deleterious effects that exceed the beneficial effect that the research was initially undertaken for, then the same can only be the case for foods that stem from the same new biotechnologies, zero risk being non existent. There are two conflicting viewpoints about the possible risks linked to genetically modified organisms: a posteriori protection (based on vigilance once the product is on the market) and an a priori protection (at present usually supported by the precaution principle). We suggest a third way, which ensures consumer safety, but doesn't hinder scientific progress. Just as there are regulations for the protection of human subjects in biomedical research and regulations for the use of drugs after they are marketed, so should such regulations be introduced in the domains of food production that use biotechnologies. We therefore suggest that the scientific community and the food-processing industry develop evaluation protocols for new foods like the ones that exist for drugs. We thus offer thirteen regulations, based on the Helsinki declaration, in order to establish these protocols. These proposals, applied to food-processing research, would enable the industry to return confidence to consumers and thus avoid the random blocking of scientific progress, which is a source of health for the greater population. PMID: 11401233 [PubMed - indexed for MEDLINE] 1992: Novartis Found Symp. 2001;236:233-9; discussion 240-1. Developing transgenic grains with improved oils, proteins and carbohydrates. Mazur BJ. DuPont Agricultural Enterprise, PO Box 80402, Wilmington, DE 19880-0402, USA. DuPont has developed cereals and oilseeds with improved proteins, carbohydrates, and oils for food, feed, and industrial applications. Products which have been or will be introduced include corn and soybeans with increased oil content, improved oil composition, increased amino acid content, altered protein content and functional qualities, altered starch composition, reduced oligosaccharide content, increased sucrose content, and combinations of these traits. These products have been developed using both mutation breeding and molecular biology-based transgenic approaches. We have also worked on improving the underlying technologies in order to accelerate product introductions. Gene discovery has been expedited through a genomics program that now has a database of more than two million sequences from a variety of plants, insects and microbes. Plant cell transformation for elite lines of crop species is being addressed through production laboratories with high throughput processes and through technology improvements. High-throughput, rapid and small-scale assays for biochemical parameters are used to identify plants carrying traits of interest. Small-scale functionality analyses, in which grains are broken down into their component parts and assayed for functional properties, indicate which seeds carry a trait of commercial value. Finally, a number of DNA marker systems are being used to accelerate trait introgression timelines. Publication Types: Review PMID: 11387983 [PubMed - indexed for MEDLINE] 1993: Nat Biotechnol. 2001 Jun;19(6):548-52. A plant-based multicomponent vaccine protects mice from enteric diseases. Yu J, Langridge WH. Department of Biochemistry and Center for Molecular Biology and Gene Therapy, Loma Linda University, Loma Linda, CA 92350, USA. Cholera toxin (CT) B and A2 subunit complementary DNAs (cDNAs) were fused to a rotavirus enterotoxin and enterotoxigenic Escherichia coli fimbrial antigen genes and transferred into potato. Immunoblot and enzyme-linked immunosorbent assay (ELISA) results indicated that the fusion antigens were synthesized in transformed tuber tissues and assembled into cholera holotoxin-like structures that retained enterocyte-binding affinity. Orally immunized mice generated detectable levels of serum and intestinal antibodies against the pathogen antigens. Elevated levels of interleukin 2 (IL2) and interferon gamma (INFgamma) detected in immunogen-challenged spleen cells from the immunized mice indicated the presence of a strong Th1 immune response to the three plant-synthesized antigens. This result was supported by flow cytometry analysis of immunized mouse spleen cells that showed a significant increase in CD4+ lymphocyte numbers. Diarrhea symptoms were reduced in severity and duration in passively immunized mouse neonates following rotavirus challenge. The results suggest that food plants can function as vaccines for simultaneous protection against infectious virus and bacterial diseases. Publication Types: Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. PMID: 11385459 [PubMed - indexed for MEDLINE] 1994: Shokuhin Eiseigaku Zasshi. 2001 Feb;42(1):24-32. A multiplex PCR method of detecting recombinant DNAs from five lines of genetically modified maize. Matsuoka T, Kuribara H, Akiyama H, Miura H, Goda Y, Kusakabe Y, Isshiki K, Toyoda M, Hino A. National Food Research Institute, Ministry of Agriculture, Forestry and Fisheries, 2-1-2, Kannondai, Tsukuba, Ibaraki 305-8642, Japan. Seven lines of genetically modified (GM) maize have been authorized in Japan as foods and feeds imported from the USA. We improved a multiplex PCR method described in the previous report in order to distinguish the five lines of GM maize. Genomic DNA was extracted from GM maize with a silica spin column kit, which could reduce experimental time and improve safety in the laboratory and potentially in the environment. We sequenced recombinant DNA (r-DNA) introduced into GM maize, and re-designed new primer pairs to increase the specificity of PCR to distinguish five lines of GM maize by multiplex PCR. A primer pair for the maize intrinsic zein gene (Ze1) was also designed to confirm the presence of amplifiable maize DNA. The lengths of PCR products using these six primer pairs were different. The Ze1 and the r-DNAs from the five lines of GM maize were qualitatively detected in one tube. The specific PCR bands were distinguishable from each other on the basis of the expected length. The r-DNA could be detected from maize samples containing 0.5% of each of the five lines of GM maize. The sensitivity would be acceptable to secure the verification of non-GMO materials and to monitor the reliability of the labeling system. Publication Types: Research Support, Non-U.S. Gov't PMID: 11383153 [PubMed - indexed for MEDLINE] 1995: FEBS Lett. 2001 May 18;497(1):50-4. Design and production of genetically modified soybean protein with anti-hypertensive activity by incorporating potent analogue of ovokinin(2-7). Matoba N, Doyama N, Yamada Y, Maruyama N, Utsumi S, Yoshikawa M. Research Institute For Food Science, Kyoto University, Uji, 611-0011, Kyoto, Japan. The potent anti-hypertensive peptide, RPLKPW, has been designed based on the structure of ovokinin(2-7). The sequence encoding this peptide was introduced into three homologous sites in the gene for soybean beta-conglycinin alpha' subunit. The native alpha' subunit as well as the modified, RPLKPW-containing alpha' subunit were expressed in Escherichia coli, recovered from the soluble fraction and then purified by ion-exchange chromatography. The RPLKPW peptide was released from recombinant RPLKPW-containing alpha' subunit after in vitro digestion by trypsin and chymotrypsin. Moreover, the undigested RPLKPW-containing alpha' subunit given orally at a dose of 10 mg/kg exerted an anti-hypertensive effect in spontaneously hypertensive rats, unlike the native alpha' subunit. These results provide evidence for the first time that a physiologically active peptide introduced into a food protein by site-directed mutagenesis could practically function in vivo even at a low dose. Publication Types: Research Support, Non-U.S. Gov't PMID: 11376661 [PubMed - indexed for MEDLINE] 1996: Arh Hig Rada Toksikol. 2001 Mar;52(1):49-59. Electrochemical biosensors for evaluation of contaminants in food. Mascini M, Palchetti I. University of Florence, Department of Public Health, Epidemiology, and Environmental Analytical Chemistry, Florence, Italy. mascini@unifi.it This paper describes the application of electrochemical disposable biosensors in food analysis, which have recently been developed in our laboratory. Disposable biosensors, based on acetylcholinesterase inhibition activity, were exploited for testing the presence of organophosphorus and carbamate pesticides in water, fruit, and vegetable samples. The paper further describes preliminary tests for the detection of genetically modified organisms and hybridisation by coupling the DNA biosensors with the polymerase chain reaction. PMID: 11370299 [PubMed - indexed for MEDLINE] 1997: Arh Hig Rada Toksikol. 2001 Mar;52(1):11-21. Quality of Life Programme--food, nutrition, and health--projects promotion. Boenke A. European Commission, Brussels, Belgium. achim.boenke@cec.eu.int The EC Quality of Life Programme (QoL), Key Action 1--Food, Nutrition & Health aims at providing a healthy, safe, and high-quality food supply leading to reinforced consumer's confidence in the safety of the European food. Key Action 1 is currently supporting several European projects investigating analytical methods for food control including sensors, risk analysis, and food safety standardisation. Their objectives range from the development and validation of prevention strategies for mycotoxin formation via the development of a communication platform for Genetically Modified Organisms (GMO), validation and standardisation of diagnostic Polymerase Chain Reaction (PCR) for food-borne pathogens, up to the evaluation of the potential cancer-preventing activity of pro- and pre-biotic ("SYNBIOTIC") combinations in human volunteers. This paper also informs on future research needs in food safety. PMID: 11370294 [PubMed - indexed for MEDLINE] 1998: J Environ Monit. 2001 Feb;3(2):26N-32N. Environmental and food safety issues of genetically modified crops. Kuiper HA. Publication Types: News PMID: 11354743 [PubMed - indexed for MEDLINE] 1999: J Allergy Clin Immunol. 2001 May;107(5):765-71. Comment in: J Allergy Clin Immunol. 2001 Oct;108(4):654. J Allergy Clin Immunol. 2001 Oct;108(4):655-6. Will genetically modified foods be allergenic? Taylor SL, Hefle SL. University of Nebraska, Food Allergy Research and Resource Program, Lincoln, USA. Foods produced through agricultural biotechnology, including such staples as corn, soybeans, canola, and potatoes, are already reaching the consumer marketplace. Agricultural biotechnology offers the promise to produce crops with improved agronomic characteristics (eg, insect resistance, herbicide tolerance, disease resistance, and climatic tolerance) and enhanced consumer benefits (eg, better taste and texture, longer shelf life, and more nutritious). Certainly, the products of agricultural biotechnology should be subjected to a careful and complete safety assessment before commercialization. Because the genetic modification ultimately results in the introduction of new proteins into the food plant, the safety, including the potential allergenicity, of the newly introduced proteins must be assessed. Although most allergens are proteins, only a few of the many proteins found in foods are allergenic under the typical circumstances of exposure. The potential allergenicity of the introduced proteins can be evaluated by focusing on the source of the gene, the sequence homology of the newly introduced protein to known allergens, the expression level of the novel protein in the modified crop, the functional classification of the novel protein, the reactivity of the novel protein with IgE from the serum of individuals with known allergies to the source of the transferred genetic material, and various physicochemical properties of the newly introduced protein, such as heat stability and digestive stability. Few products of agricultural biotechnology (and none of the current products) will involve the transfer of genes from known allergenic sources. Applying such criteria provides reasonable assurance that the newly introduced protein has limited capability to become an allergen. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 11344340 [PubMed - indexed for MEDLINE] 2000: Trends Biotechnol. 2001 Apr;19(4):130-1. The biotechnology industry's Frankensteinian creation. Miller HI. Publication Types: Letter PMID: 11335992 [PubMed - indexed for MEDLINE] 2001: Int J Parasitol. 2001 May 1;31(5-6):621-7. Bacterial symbiosis and paratransgenic control of vector-borne Chagas disease. Beard CB, Dotson EM, Pennington PM, Eichler S, Cordon-Rosales C, Durvasula RV. Division of Parasitic Diseases, Centers for Disease Control and Prevention, Atlanta, GA, USA. cbeard@cdc.gov The triatomine vectors of Chagas disease are obligate haematophagous insects, feeding on vertebrate blood throughout their entire developmental cycle. As a result of obtaining their nutrition from a single food source, their diet is devoid of certain vitamins and nutrients. Consequently, these insects harbour populations of bacterial symbionts within their intestinal tract, which provide the required nutrients that are lacking from their diet. We have isolated and characterised symbiont cultures from various triatomine species and developed a method for genetically transforming them. We can then reintroduce them into their original host species, thereby producing stable paratransgenic insects in which we are able to express heterologous gene products. Using this methodology, we have generated paratransgenic Rhodnius prolixus that are refractory for infection with Trypanosoma cruzi. Two examples of potentially refractory genes are currently being expressed in paratransgenic insects. These include the insect immune peptide cecropin A and active single chain antibody fragments. We have also developed an approach that would allow introduction of genetically modified bacterial symbionts into natural populations of Chagas disease vectors. This approach utilises the coprophagic behaviour of these insects, which is the way in which the symbionts are transmitted among bug populations in nature. The production and ultimate release of transgenic or paratransgenic insects for public health applications is potentially very promising but also worthy of much careful consideration with respect to environmental, political, and human safety concerns. Publication Types: Review PMID: 11334952 [PubMed - indexed for MEDLINE] 2002: Risk Anal. 2001 Feb;21(1):189-98. Limits of knowledge and the limited importance of trust. Sjöberg L. Center for Risk Research, Stockholm School of Economics, Sweden. pls@hhs.se Perceived risk and related attitudes have been implicated as major factors in many of the difficult policy problems that face modern society (nuclear power, genetically modified food, etc). Experts often argue that no or very small risks are involved; people are still worried. Why? The standard answer is lack of trust. Data on trust and risk perception, however, point to only a weak relationship between the two (r approximately 0.3). It is suggested here that the reason for the surprisingly minor importance of trust is that people believe that there are clear limits to how much science and experts know. Results are presented from studies of risk perception of the public, experts, and politicians. Politicians and members of the public believe that there are many unknown effects of technology and such beliefs were strongly related to their perceived risk. Experts on nuclear waste, on the other hand, seemed to believe that little is unknown in their field of expertise. Regression analyses of risk perception showed the unknown-effects factor to be a more important explanatory factor than trust for the public and politicians. PMID: 11332547 [PubMed] 2003: Proc Natl Acad Sci U S A. 2001 May 22;98(11):6511-5. Epub 2001 May 1. Tomato Ve disease resistance genes encode cell surface-like receptors. Kawchuk LM, Hachey J, Lynch DR, Kulcsar F, van Rooijen G, Waterer DR, Robertson A, Kokko E, Byers R, Howard RJ, Fischer R, Prufer D. Lethbridge Research Centre, Agriculture and Agri-Food Canada, P.O. Box 3000, Lethbridge, AB, Canada T1J 4B1. kawchuk@em.agr.ca In tomato, Ve is implicated in race-specific resistance to infection by Verticillium species causing crop disease. Characterization of the Ve locus involved positional cloning and isolation of two closely linked inverted genes. Expression of individual Ve genes in susceptible potato plants conferred resistance to an aggressive race 1 isolate of Verticillium albo-atrum. The deduced primary structure of Ve1 and Ve2 included a hydrophobic N-terminal signal peptide, leucine-rich repeats containing 28 or 35 potential glycosylation sites, a hydrophobic membrane-spanning domain, and a C-terminal domain with the mammalian E/DXXXLphi or YXXphi endocytosis signals (phi is an amino acid with a hydrophobic side chain). A leucine zipper-like sequence occurs in the hydrophobic N-terminal signal peptide of Ve1 and a Pro-Glu-Ser-Thr (PEST)-like sequence resides in the C-terminal domain of Ve2. These structures suggest that the Ve genes encode a class of cell-surface glycoproteins with receptor-mediated endocytosis-like signals and leucine zipper or PEST sequences. Publication Types: Research Support, Non-U.S. Gov't PMID: 11331751 [PubMed - indexed for MEDLINE] 2004: Nat Biotechnol. 2001 May;19(5):470-4. Comment in: Nat Biotechnol. 2001 Sep;19(9):811. Overexpression of petunia chalcone isomerase in tomato results in fruit containing increased levels of flavonols. Muir SR, Collins GJ, Robinson S, Hughes S, Bovy A, Ric De Vos CH, van Tunen AJ, Verhoeyen ME. Unilever Research, Colworth House, Sharnbrook, Bedfordshire MK44 1LQ, UK. Tomatoes are an excellent source of the carotenoid lycopene, a compound that is thought to be protective against prostate cancer. They also contain small amounts of flavonoids in their peel ( approximately 5-10 mg/kg fresh weight), mainly naringenin chalcone and the flavonol rutin, a quercetin glycoside. Flavonols are very potent antioxidants, and an increasing body of epidemiological data suggests that high flavonoid intake is correlated with a decreased risk for cardiovascular disease. We have upregulated flavonol biosynthesis in the tomato in order to generate fruit with increased antioxidant capacity and a wider range of potential health benefit properties. This involved transformation of tomato with the Petunia chi-a gene encoding chalcone isomerase. Resulting transgenic tomato lines produced an increase of up to 78 fold in fruit peel flavonols, mainly due to an accumulation of rutin. No gross phenotypical differences were observed between high-flavonol transgenic and control lines. The phenotype segregated with the transgene and demonstrated a stable inheritance pattern over four subsequent generations tested thus far. Whole-fruit flavonol levels in the best of these lines are similar to those found in onions, a crop with naturally high levels of flavonol compounds. Processing of high-flavonol tomatoes demonstrated that 65% of flavonols present in the fresh fruit were retained in the processed paste, supporting their potential as raw materials for tomato-based functional food products. PMID: 11329019 [PubMed - indexed for MEDLINE] 2005: Nat Biotechnol. 2001 May;19(5):417-8. Comment on: Nat Biotechnol. 2001 May;19(5):466-9. Improving rice yields--ironing out the details. Guerinot ML. Publication Types: Comment News PMID: 11329003 [PubMed - indexed for MEDLINE] 2006: Nat Biotechnol. 2001 May;19(5):405. The limits of GMO detection. Kay S, Van den Eede G. Publication Types: Letter PMID: 11328998 [PubMed - indexed for MEDLINE] 2007: Toxicol Lett. 2001 Mar 31;120(1-3):181-6. Determination of protein allergenicity: studies in mice. Dearman RJ, Kimber I. Syngenta Central Toxicology Laboratory, Alderley Park, Cheshire SK10 4TJ, Macclesfield, UK. rebecca.dearman@syngenta.com There is a need to identify and characterize the allergenic potential of novel proteins introduced into genetically-modified crop plants. Although several approaches have already been described, none of these measures directly the ability of proteins to cause allergic sensitization. For this reason there has been a growing interest in the development of suitable animal models. This article describes experience to date with a method based upon assessment of serological (IgG and IgE antibody) responses induced in BALB/c strain mice by proteins. Comparisons have been made between intraperitoneal (i.p.) administration and exposure by gavage using both allergenic and non-allergenic proteins. The available data indicate that responses provoked by i.p. exposure permit the identification of proteins that have the inherent potential to induce IgE antibody production and allergic sensitization. Moreover, this approach also provides a rank order of proteins with respect to allergenic potency that apparently reflects what is known of their relative sensitizing activity in humans. By comparison, oral exposure of mice by gavage is somewhat less sensitive. On this basis it is proposed that the inherent sensitizing potential of novel proteins can be evaluated as a function of IgE antibody responses stimulated by parenteral (i.p.) exposure of BALB/c mice. PMID: 11323176 [PubMed - indexed for MEDLINE] 2008: Toxicol Lett. 2001 Mar 31;120(1-3):165-70. Food allergy: what are the issues? Kimber I, Dearman RJ. Syngenta Central Toxicology Laboratory, Alderley Park, Cheshire SK10 4TJ, Macclesfield, UK. ian.kimber@syngenta.com With a growing interest in the development of genetically modified crop plants there is a need for appropriate approaches to safety assessment. Among the issues that have to be addressed is consideration of whether the products of novel genes have the potential to cause allergic sensitization. Resulting from a collaboration between the International Food Biotechnology Council and the International Life Sciences Institute recommendations have been made for a step-wise approach to the assessment of allergenic potential based upon considerations of serological identity, and sequence or structural homology, with known allergens and examination of the stability of the test protein in a simulated gastric fluid. In parallel there has been interest in the development of animal models, which would permit a more direct evaluation of potential allergenic activity. Progress in these areas is reviewed briefly in the context of what is known of food allergy and some of the important issues, which must be addressed in designing safety assessment strategies identified. Publication Types: Review PMID: 11323174 [PubMed - indexed for MEDLINE] 2009: J Agric Food Chem. 2000 Dec;48(12):5936-45. Compositional analysis of tubers from insect and virus resistant potato plants. Rogan GJ, Bookout JT, Duncan DR, Fuchs RL, Lavrik PB, Love SL, Mueth M, Olson T, Owens ED, Raymond PJ, Zalewski J. Monsanto Company, 700 Chesterfield Village Parkway, St. Louis, Missouri 63198, USA. j.rogan@monsanto.com Genetically modified potato plants that are resistant to the Colorado potato beetle, plus either the potato leaf roll virus or potato virus Y, have recently been commercialized. As part of the safety assessment for plants produced by modern biotechnology, the composition of the food/feed must be compared to that of the food/feed produced by an equivalent plant variety from a conventional source. The composition of important nutritional and antinutritional factors in tubers produced by virus- and insect-resistant potato plants were compared to tubers produced by conventional potato plants. Key nutritional, quality, and antinutritional components measured were total solids, vitamin C, dextrose, sucrose, soluble protein, and glycoalkaloids. Proximate analyses included fat, ash, calories, total protein, and crude fiber. Minor nutrients measured were vitamin B6, niacin, copper, magnesium, potassium, and amino acids. The results from these analyses confirm that tubers produced by insect- and virus-protected varieties are substantially equivalent to tubers produced by conventional potato varieties. PMID: 11312768 [PubMed - indexed for MEDLINE] 2010: Int Arch Allergy Immunol. 2001 Jan-Mar;124(1-3):51-4. Reduction in allergenicity of grass pollen by genetic engineering. Bhalla PL, Swoboda I, Singh MB. Plant Molecular Biology and Biotechnology Laboratory, Institute of Land and Food Resources, University of Melbourne, Parkville, Australia. p.bhalla@landfood.unimelb.edu.au BACKGROUND: Hay fever and allergic asthma triggered by grass pollen allergens affect approximately 20% of the population in cool temperate climates. Ryegrass is the dominant source of allergens due to its prodigious airborne pollen production. Lol p 5 or group 5 is among the most important and widespread grass pollen allergen because it reacts with IgE antibodies of more than 90% of grass pollen-allergic patients, contains most of the grass pollen-specific IgE epitopes and elicits strong biological responses. Significant efforts have been made in developing diagnostic and therapeutic reagents for designing new and more effective immunotherapeutic strategies for treatment of allergic diseases. An alternative approach to this problem could be to reduce the amount of allergen content in the source plant. METHODS: High velocity microprojectile bombardment was used to genetically engineer ryegrass. Antisense construct targeted to one of major allergen, Lol p 5, was introduced. The expression of antisense RNA was regulated by a pollen-specific promoter. Pollen was analysed for IgE reactivity. RESULTS: Analysis of proteins with allergen-specific monoclonal and polyclonal antibodies did not detect Lol p 5 in the transgenic pollen. The transgenic pollen showed remarkably reduced allergenicity as reflected by low IgE binding capacity of pollen extract as compared to control pollen. The transgenic ryegrass plants in which Lol p 5 gene expression is perturbed showed normal fertile pollen development. CONCLUSIONS: Our studies showed that it is possible to selectively 'switch off' allergen production in pollen of ryegrass demonstrating feasibility of genetic engineering of plants for reduced allergenicity. Copyright 2001 S. Karger AG, Basel Publication Types: Research Support, Non-U.S. Gov't PMID: 11306924 [PubMed - indexed for MEDLINE] 2011: EMBO Rep. 2001 Apr;2(4):256-8. Comment in: EMBO Rep. 2001 Jul;2(7):545-8. Genetically modified crops: hope for developing countries? The current GM debate widely ignores the specific problems of farmers and consumers in the developing world. Herrera-Estrella L, Alvarez-Morales A. Departamento de Ingeniería Genética de plantas, Centro de Investigación y Estudios Avanzados del IPN in Irapuato, Guanajuato, México. lherrera@ira.cinvestav.mx PMID: 11306538 [PubMed - indexed for MEDLINE] 2012: Biosci Biotechnol Biochem. 2001 Feb;65(2):383-8. A transgenic apple callus showing reduced polyphenol oxidase activity and lower browning potential. Murata M, Nishimura M, Murai N, Haruta M, Homma S, Itoh Y. Department of Nutrition and Food Science, Ochanomizu University, Tokyo, Japan. murata@cc.ocha.ac.jp Polyphenol oxidase (PPO) is responsible for enzymatic browning of apples. Apples lacking PPO activity might be useful not only for the food industry but also for studies of the metabolism of polyphenols and the function of PPO. Transgenic apple calli were prepared by using Agrobacterium tumefaciens carrying the kanamycin (KM) resistant gene and antisense PPO gene. Four KM-resistant callus lines were obtained from 356 leaf explants. Among these transgenic calli, three calli grew on the medium containing KM at the same rate as non-transgenic callus on the medium without KM. One callus line had an antisense PPO gene, in which the amount and activity of PPO were reduced to half the amount and activity in non-transgenic callus. The browning potential of this line, which was estimated by adding chlorogenic acid, was also half the browning potential of non-transgenic callus. Publication Types: Research Support, Non-U.S. Gov't PMID: 11302173 [PubMed - indexed for MEDLINE] 2013: Curr Biol. 2001 Mar 20;11(6):R201. GM crops under new US scrutiny. Bonetta L. Publication Types: News PMID: 11301260 [PubMed - indexed for MEDLINE] 2014: Curr Biol. 2001 Mar 20;11(6):R199-200. Europe opens the door to GM crops. Williams N. Publication Types: News PMID: 11301259 [PubMed - indexed for MEDLINE] 2015: Allergy. 2001;56 Suppl 67:61-3. How to make foods safer--genetically modified foods. Moseley BE. Reading, Berkshire, UK. It is the responsibility of companies developing genetically modified foods, and of regulatory authorities that approve their marketing, to ensure that they are at least as safe as the traditional foods they are intended to replace in the diet. This requires that any novel material introduced into the food material should not be allergenic. If the novel gene has come from an allergenic source, e.g. nuts, it is necessary to demonstrate using immunological procedures applied to the IgE fractions of pooled sera from individuals with confirmed allergies that the novel protein is non-allergenic. When the novel gene is from a non-allergenic source then it is necessary to demonstrate lack of significant amino acid sequence homology to known allergens together with sensitivity to food manufacturing and digestive processes. Consumer confidence in genetically modified foods would be significantly improved if hypoallergenic varieties of crops and food products that are currently allergenic could be developed. Techniques such as antisense technology and single site amino acid substitution have been shown to have such potential. Publication Types: Review PMID: 11298012 [PubMed - indexed for MEDLINE] 2016: Med Law. 2000;19(4):703-11. Biotechnology-based foods: is there a third way between the precaution principle and an overly enthusiastic dissemination of GMO? Meningaud JP, Moutel G, Herv C. Laboratoire d'ethique médicale et de santé publique, Faculté de médecine Necker-Université, Paris. The demand for consumer safety with regard to the food-processing industry is becoming, legitimately, more and more urgent. If ingested drugs can carry deleterious effects that exceed the beneficial effect that the research was initially undertaken for, then the same can only be the case for foods that stem from the same new biotechnologies, zero risk being non existent. PMID: 11289641 [PubMed - indexed for MEDLINE] 2017: Med Anthropol Q. 2001 Mar;15(1):29-30. Comment on: Med Anthropol Q. 2001 Mar;15(1):9-19; discussion 20-1. Health, environment, and transgenic agriculture. Diamond J. Publication Types: Comment PMID: 11288614 [PubMed - indexed for MEDLINE] 2018: Med Anthropol Q. 2001 Mar;15(1):22-5. Comment on: Med Anthropol Q. 2001 Mar;15(1):9-19; discussion 20-1. Toward a political economy of opinion formation on genetically modified foods. Rosset PM. Publication Types: Comment PMID: 11288612 [PubMed - indexed for MEDLINE] 2019: Sci Am. 2001 Apr;284(4):64-5. Does the world need GM foods? No. Interview by Sasha Nemecek. Mellon M. Publication Types: Interview PMID: 11285823 [PubMed - indexed for MEDLINE] 2020: Sci Am. 2001 Apr;284(4):62-3. Does the world need GM foods? Yes. Interview by Sasha Nemecek. Horsch RB. Publication Types: Interview PMID: 11285822 [PubMed - indexed for MEDLINE] 2021: Sci Am. 2001 Apr;284(4):60-1. The risks on the table. Hopkin K. PMID: 11285821 [PubMed - indexed for MEDLINE] 2022: Sci Am. 2001 Apr;284(4):52-7. Seeds of concern. Brown K. Publication Types: Review PMID: 11285820 [PubMed - indexed for MEDLINE] 2023: Nature. 2001 Mar 29;410(6828):503. Critics claim 'sight-saving' rice is over-rated. Schnapp N, Schiermeier Q. Publication Types: News PMID: 11279451 [PubMed - indexed for MEDLINE] 2024: Nature. 2001 Mar 29;410(6828):501. Americans perplexed by GM food. Lok C. Publication Types: News PMID: 11279448 [PubMed - indexed for MEDLINE] 2025: CMAJ. 2001 Mar 20;164(6):853. Feds' approach to genetically modified products criticized. Sibbald B. Publication Types: News PMID: 11276555 [PubMed - indexed for MEDLINE] 2026: Mol Cells. 2001 Feb 28;11(1):7-12. Generation of a murine single chain Fv (scFv) antibody specific for cucumber mosaic virus (CMV) using a phage display library. Chae JS, Choi JK, Lim HT, Cha SH. Division of Food Science & Biotechnology, College of Agriculture & Life Sciences, Kangwon National University, Chunchon, Korea. With the long-term goal of generating CMV-resistant transgenic plants using antibody genes, a single-chain variable fragment (scFv) antibody that binds to the cucumber mosaic virus was isolated from a scFv phage display library by four rounds of affinity selection with CMV-Mf as an antigen. The scFv has the identical binding specificity to CMV as a monoclonal antibody that is generated by the hybridoma fusion technique, and recognized purified preparations of CMV isolates belonging to either subgroup I or II in immunoblotting. The nucleotide sequences of the recombinant antibody showed that a heavy chain variable region (V(H)) gene belonged to the VH3 subgroup and the kappa light chain variable region (V kappa) came from the Vkappa4 subgroup. Our results demonstrate that the scFv phage display library, an alternative approach to the traditional hybridoma fusion technique, has a potential applicability in the study of plant virus and plant pathology. Publication Types: Research Support, Non-U.S. Gov't PMID: 11266124 [PubMed - indexed for MEDLINE] 2027: J Agric Food Chem. 2001 Feb;49(2):652-7. Control of enzymatic browning in potato (Solanum tuberosum L.) by sense and antisense RNA from tomato polyphenol oxidase. Coetzer C, Corsini D, Love S, Pavek J, Tumer N. Biotechnology Center for Agriculture and the Environment and Department of Plant Pathology, Rutgers University, Cook College, P.O. Box 231, New Brunswick, New Jersey 08903-0231, USA. Polyphenol oxidase (PPO) activity of Russet Burbank potato was inhibited by sense and antisense PPO RNAs expressed from a tomato PPO cDNA under the control of the 35S promoter from the cauliflower mosaic virus. Transgenic Russet Burbank potato plants from 37 different lines were grown in the field. PPO activity and the level of enzymatic browning were measured in the tubers harvested from the field. Of the tubers from 28 transgenic lines that were sampled, tubers from 5 lines exhibited reduced browning. The level of PPO activity correlated with the reduction in enzymatic browning in these lines. These results indicate that expression of tomato PPO RNA in sense or antisense orientation inhibits PPO activity and enzymatic browning in the major commercial potato cultivar. Expression of tomato PPO RNA in sense orientation led to the greatest decrease in PPO activity and enzymatic browning, possibly due to cosuppression. These results suggest that expression of closely related heterologous genes can be used to prevent enzymatic browning in a wide variety of food crops without the application of various food additives. PMID: 11262007 [PubMed - indexed for MEDLINE] 2028: Bull Acad Natl Med. 2000;184(7):1477-86; discussion 1487-90. [Endocrine disruption agents: environment, health, public policies, and the precautionary principle] [Article in French] Vandelac L. Département de sociologie Institut des sciences de l'environnement de l'Université du Québec à Montréal. The already substantial body of evidence and growing web of suspicions as to the scale and severity of the cascade effects of endocrine disrupters (related to persistent organic pollutants or POPs) on the health of ecosystems and humans have sparked such concern that in June 1998, representatives of 94 countries meeting in Montreal under the aegis of UNEP signed a draft international agreement to phase out the most harmful POPs. Related to particular persistent organic pollutants--toxic semi-volatile and persistent chemical compounds now found everywhere in the environment, such as BPCs, organochlorine pesticides, dioxins and furans, that build up in the bodies of organisms that consume other contaminated organisms along the food chain--endocrine disrupters are strongly suspected of affecting the health of animals and adversely impacting the health, fertility and even intellectual faculties of humans. For example, very low-level exposure to some POPs is associated with some hormone-dependent cancers, damage to the central and peripheral nervous systems, impaired immune system function, reproductive disorders and developmental disruptions in newborns and infants, who can be affected in utero or through breast-feeding. Considering the extreme complexity of the scientific and socio-economic effects of POP-related endocrine disrupters, there are those who, advocate a wait-and-see approach, claiming that there is not enough formal scientific proof. There are others who use the available evidence to advance the research, press for bans on incriminated substances and look for global, integrated and viable alternatives. And there are other still who, with careless disregard for the Precautionary Principle, are quite prepared to talk about the perverse effects of POPs in order to justify the increased use of artificial means of reproduction or the replacement of chemical pesticides by pest-resistant genetically modified organisms (GMOs), thereby opening the door to "solutions" that are potentially more biologically and ethically dangerous than the problems they purport to remedy. This paper provides an overview of the current understanding of the main sources and suspected effects of POPs on animal life and human health, explores the complexity of the scientific, economic and political issues involved in any international process to do away with the incriminated products, discusses the risks and perverse consequences of some of the proposed alternatives, and stresses the importance, in the light of these risks and consequences, of placing renewed emphasis on public and environmental health approaches based on the Precautionary Principle. Publication Types: English Abstract Review PMID: 11261252 [PubMed - indexed for MEDLINE] 2029: Nat Rev Genet. 2001 Mar;2(3):217-22. Scientific perspectives on regulating the safety of genetically modified foods. Gasson M, Burke D. Food Science Division, Institute of Food Research, Norwich Research Park, Colney, Norwich NR4 7UA, UK. Regulation is often seen as the dull end of science. The recent storm over the introduction of genetically modified foods and the calls to regulate their consumption have had a negative effect on development of the science. Assuring the safety of genetically modified foods might raise questions where existing scientific data is limited and underline the need for further research. Publication Types: Review PMID: 11256073 [PubMed - indexed for MEDLINE] 2030: Biotechnol Genet Eng Rev. 2000;17:327-52. Genetically modified food crops: current concerns and solutions for next generation crops. Daniell H. Department of Molecular Biology and Microbiology, 12722 Research Parkway, University of Central Florida, Orlando, FL 32826-3227, USA. daniell@mail.ucf.edu Publication Types: Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S. Review PMID: 11255672 [PubMed - indexed for MEDLINE] 2031: EMBO Rep. 2001 Jan;2(1):10-1. A doorman to keep foreign genes out. A new genetic barrier can protect corn from unwanted genetic modification. Brower V. vickibrower@cs.com PMID: 11252714 [PubMed - indexed for MEDLINE] 2032: Transgenic Res. 2001;10(1):1-12. Antibody-based resistance to plant pathogens. Schillberg S, Zimmermann S, Zhang MY, Fischer R. FraunhoferAbteilung für Molekulare Biotechnologie, IUCT, Grafschaft, Schmallenberg, Germany. schillberg@bio1.rwth-aachen.de Plant diseases are a major threat to the world food supply, as up to 15% of production is lost to pathogens. In the past, disease control and the generation of resistant plant lines protected against viral, bacterial or fungal pathogens, was achieved using conventional breeding based on crossings, mutant screenings and backcrossing. Many approaches in this field have failed or the resistance obtained has been rapidly broken by the pathogens. Recent advances in molecular biotechnology have made it possible to obtain and to modify genes that are useful for generating disease resistant crops. Several strategies, including expression of pathogen-derived sequences or anti-pathogenic agents, have been developed to engineer improved pathogen resistance in transgenic plants. Antibody-based resistance is a novel strategy for generating transgenic plants resistant to pathogens. Decades ago it was shown that polyclonal and monoclonal antibodies can neutralize viruses, bacteria and selected fungi. This approach has been improved recently by the development of recombinant antibodies (rAbs). Crop resistance can be engineered by the expression of pathogen-specific antibodies, antibody fragments or antibody fusion proteins. The advantages of this approach are that rAbs can be engineered against almost any target molecule, and it has been demonstrated that expression of functional pathogen-specific rAbs in plants confers effective pathogen protection. The efficacy of antibody-based resistance was first shown for plant viruses and its application to other plant pathogens is becoming more established. However, successful use of antibodies to generate plant pathogen resistance relies on appropriate target selection, careful antibody design, efficient antibody expression, stability and targeting to appropriate cellular compartments. Publication Types: Review PMID: 11252378 [PubMed - indexed for MEDLINE] 2033: Curr Opin Mol Ther. 2000 Feb;2(1):80-6. Development of oral vaccines for human use. Fooks AR. Centre for Applied Microbiology and Research (CAMR), Salisbury, Wiltshire SP4 0JG, UK. anthony.fooks@camr.org.uk In developed and developing countries, oral vaccine formulations that elicit protection at mucosal surfaces are attractive vaccine candidates. Research has shown that vaccine delivery using either viral or non-viral vector delivery of heterologous proteins via the oral route is highly effective. Improvements in non-viral vector uptake, specific targeting, antigen presentation and antigen release times will be required to overcome differences in the immune response following delivery. In contrast, the use of plant and animal viruses as vectors provides an effective method for targeted gene delivery. Recently, recombinant plant viruses grown in plants or transgenic plants have been proposed as edible vaccines for human use. Food plants offer many advantages as affordable, oral vaccines, especially for use in developing countries. Further research is necessary to develop strategies to improve immunological memory following oral vaccination and to avoid immunological tolerance in the host. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 11249655 [PubMed - indexed for MEDLINE] 2034: Nature. 2001 Feb 15;409(6822):749. Call for tighter controls on transgenic foods. Spurgeon D. Publication Types: News PMID: 11236963 [PubMed - indexed for MEDLINE] 2035: Nat Biotechnol. 2001 Mar;19(3):183-4. US Presidential transition prompts biotech policy frenzy. Fox JL. Publication Types: News PMID: 11231515 [PubMed - indexed for MEDLINE] 2036: Arch Insect Biochem Physiol. 2000 Dec;45(4):175-9. Modulation of a lectin insecticidal activity by carbohydrates. Triguéros V, Wang M, Père D, Paquereau L, Chavant L, Fournier D. Université Paul Sabatier, Laboratoire de Synthèse et Physicochimie des Molécules d'Intérêt Biologique, UMR CNRS 5068, Groupe de Biochimie des Protéines, Toulouse, France. Lectins from plants present an insecticidal activity most probably through their carbohydrate binding properties; as a consequence, their toxicity should vary with the presence of a competitive sugar in the ingested food. In order to test this hypothesis, we performed competition experiments between insecticidal activity and carbohydrate binding. For this purpose, we used a lectin from Lathyrus ochrus and the specific carbohydrate for this protein, glucose. In toxicological tests with Drosophila melanogaster, we observed a decrease of lectin toxicity when glucose was added to the larva-rearing medium. This result suggests that the toxicity of the lectin is correlated to its ability to bind sugar in the insect digestive tract and stresses the importance of sugar composition of the nutriment used for toxicological testing of lectins or in genetically modified plants. PMID: 11223937 [PubMed - indexed for MEDLINE] 2037: Biologist (London). 2000 Jun;47(3):113. Comment on: Biologist (London). 2000 Feb;47(1):7-10. Genetically modified crops and factual information. Griffiths M. Publication Types: Comment Letter PMID: 11190239 [PubMed - indexed for MEDLINE] 2038: Biologist (London). 2000 Apr;47(2):81-4. Is there a future for GMOs? Beringer JE. School of Biological Sciences, University of Bristol, Woodland Road, Bristol, BS8 1UG. Despite strict regulation and a clean safety record, research and development of genetically modified (GM) crops and other organisms has been confronted with tremendous public hostility. Why has this happened, and how can scientists try to guide the debate into more rational channels? The answers may determine the future of GM technology and our ability to provide for a growing world population. Publication Types: Review PMID: 11190234 [PubMed - indexed for MEDLINE] 2039: Biologist (London). 2000 Apr;47(2):74-6. GM frenzy: a lesson in communication. Dixon B. Hysteria rather than rationality has characterised the debate over genetically modified foods, which erupted in Britain in 1998 and has since spread to other countries. This episode holds wider lessons for biologists, politicians and all concerned with the prudent assessment and application of novel technology. Publication Types: Review PMID: 11190232 [PubMed - indexed for MEDLINE] 2040: Science. 2000 Dec 8;290(5498):1867. Toxicology. Panel urges further study of biotech corn. Kaiser J. Publication Types: News PMID: 11187032 [PubMed - indexed for MEDLINE] 2041: Science. 2000 Oct 20;290(5491):457-9. Genetic technologies. Bioengineered food--safety and labeling. Goldman KA. Georgetown University Law Center, Washington, DC 20001, USA. goldmank@law.georgetown.edu The safety and labeling of genetically engineered foods are two areas that have elicited considerable public concern and debate. This Policy Forum provides a legal analysis of these issues in the context of two bills that have been recently proposed in The U.S. Congress, the Genetically Engineered Food Safety Act and the Genetically Engineered Food Right to Know Act. Most transgenic components of foods currently on the market are plant-incorporated protectants or their inert ingredients. Therefore, they have been evaluated for safety by the Environmental Protection Agency (as well as the Food and Drug Administration), and their disclosure in labeling should not be required. If plant-incorporated protectants are considered safer than chemical pesticides, and chemical pesticides do not have to be disclosed in labels, then bioengineered foods should not be subject to stricter regulation, nor should they be required to be labeled. The two bills are inconsistent, in many respects, with well-established principles of food regulation. PMID: 11183767 [PubMed - indexed for MEDLINE] 2042: Trends Biotechnol. 2000 Oct;18(10):404-5. Transgenic crops from another perspective. Auberson L. Agency BATS, Basel, Switzerland. auberson@bats.ch Publication Types: Congresses PMID: 11183126 [PubMed - indexed for MEDLINE] 2043: Nat Biotechnol. 2001 Feb;19(2):117-20. Betting on biogenerics. Dove A. PMID: 11175723 [PubMed - indexed for MEDLINE] 2044: Nat Biotechnol. 2001 Feb;19(2):93. 'Twixt cup and lip--biotechnology and resource-poor farmers. Tripp R. Publication Types: Editorial PMID: 11175684 [PubMed - indexed for MEDLINE] 2045: J Agric Food Chem. 2001 Jan;49(1):395-401. Prolamin aggregation, gluten viscoelasticity, and mixing properties of transgenic wheat lines expressing 1Ax and 1Dx high molecular weight glutenin subunit transgenes. Popineau Y, Deshayes G, Lefebvre J, Fido R, Tatham AS, Shewry PR. Unité de Biochimie et de Technologie des Protéines and Unité de Physicochimie des Macromolécules, INRA, B.P. 71627, Rue de la Géraudière, 44316 Nantes Cedex 03, France. popineau@nantes.inra.fr The composition of high molecular weight (HMW) subunits of glutenin determines the gluten strength and influences the baking quality of bread wheat. Here, the effect of transgenes coding for subunits 1Ax1 and 1Dx5 was studied in two near-isogenic wheat lines differing in their HMW subunit compositions and mixing properties. The subunits encoded by the transgenes were overexpressed in the transformed lines and accounted for 50-70% of HMW subunits. Overexpression of 1Ax1 and 1Dx5 subunits modified glutenin aggregation, but glutenin properties were much more affected by expression of the 1Dx5 transgene. This resulted in increased cross-linking of glutenin polymers. In dynamic assay, the storage and loss moduli of hydrated glutens containing 1Dx5 transgene subunits were considerably enhanced, whereas expression of the 1Ax1 transgene had a limited effect. The very high strength of 1Dx5 transformed glutens resulted in abnormal mixing properties of dough. These results are discussed with regard to glutenin subunit and glutenin polymer structures. Publication Types: Research Support, Non-U.S. Gov't PMID: 11170604 [PubMed - indexed for MEDLINE] 2046: J Biotechnol. 2001 Feb 13;85(2):227-39. Biosafety: future priorities for research in health care. Doblhoff-Dier O, Collins CH. Institute for Applied Microbiology, University of Agricultural Sciences, Nussdorfer Lände 11, A-1190, Vienna, Austria. doblhoff@boku.ac.at Currently the public interest in biosafety issues has focussed on the discussions surrounding the use of genetically modified organisms, very specifically on the use of transgenic plants in agriculture. Although many of the questions raised in connection with genetically modified organisms are of legitimate scientific interest, attention should be drawn back to a number of other more classical biosafety research areas, namely the problem of control of new and reemerging infectious diseases, the need for new vaccines, control of transport and routes of dissemination, biosafety information exchange and networking, where research results are dearly needed. In the area of modern biotechnology new applications such as gene therapy and transgenic animals will be on the list of future priorities for biosafety related activities and research. PMID: 11165365 [PubMed - indexed for MEDLINE] 2047: J Biotechnol. 2001 Jan 23;85(1):49-56. Expression and purification of polyhistidine-tagged firefly luciferase in insect cells--a potential alternative for process scale-up. Michel P, Torkkeli T, Karp M, Oker-Blom C. VTT Biotechnology and Food Research, FIN-02044 VTT, Espoo, Finland. The coleopteran firefly, Photinus pyralis, luciferase was produced in lepidopteran Trichoplusia ni insect cells using a baculovirus expression vector. The recombinant protein was equipped with a polyhistidine affinity tag at the carboxyl terminus and purified by immobilized metal-ion affinity chromatography in combination with an expanded bed adsorption system. This approach enabled an efficient, one-step purification protocol of a genetically modified luciferase with properties similar to those of the authentic counterpart. According to light emission measurements, the final yield of highly purified protein was 23 mg l(-1) of cell culture. In addition, no specific interaction of interfering substances, such as, ATP, adenylate kinase, nucleoside diphosphokinase, as well as, creatine kinase of the final preparation were identified. Together, the results presented here clearly show that the baculovirus expression system in combination with immobilized metal-ion affinity chromatography is a potential strategy for process scale-up of polyhistidine tagged insect luciferase. Publication Types: Research Support, Non-U.S. Gov't PMID: 11164962 [PubMed - indexed for MEDLINE] 2048: Trends Biotechnol. 2001 Feb;19(2):67-73. Metal-binding proteins and peptides in bioremediation and phytoremediation of heavy metals. Mejáre M, Bülow L. Dept of Pure and Applied Biochemistry, Centre for Chemistry and Chemical Engineering, P.O. Box 124, S-221 00, Lund, Sweden. The expression of metal-binding proteins or peptides in microorganisms and plants in order to enhance heavy metal accumulation and/or tolerance has great potential. Several different peptides and proteins have been explored. This review focuses on cadmium (Cd) because of the significant importance of this metal and because of its global presence in many food materials. Publication Types: Review PMID: 11164556 [PubMed - indexed for MEDLINE] 2049: Trends Biotechnol. 2001 Feb;19(2):43-8. Irish public perceptions and attitudes to modern biotechnology: an overview with a focus on GM foods. Morris SH, Adley CC. Irish Visiting Researcher, Center for Safe Food, Department of Plant Agriculture, University of Guelph, Ontario, N1G 2W1, Guelph, Canada. morris@uoguelph.ca This article summarizes the current situation pertaining to modern biotechnology in Ireland, with a particular focus on genetically modified (GM) crops. It briefly examines some important results of the major national surveys carried out in Ireland since 1989, highlights the recent upsurge in media (newspaper) coverage of GM related stories in three Irish opinion leader publications and it allows for an insight into the Irish public's relationship with modern biotechnology. PMID: 11164552 [PubMed - indexed for MEDLINE] 2050: Trends Plant Sci. 2001 Jan;6(1):14-7. Genetic manipulation of carotenoid biosynthesis: strategies, problems and achievements. Sandmann G. Biosynthesis Group, Botanical Institute 213, Goethe Universität Frankfurt, PO Box 111932, 60054, Frankfurt, Germany. sandmann@em.uni-frankfurt.d400.de Carotenoids, some of which are provitamin A, have a range of diverse biological functions and actions, especially in relation to human health. For example, carotenoids are known to be crucial for normal vision and have been associated with reducing the risk of several degenerative diseases including cancer. The putative advantage of modifying and engineering the carotenoid biosynthetic pathways is obvious: to provide sources for the isolation of desired carotenoids or to generate food plants with increased carotenoid content. This article reviews the studies of carotenoid production in heterologous microorganisms and the engineering of crop plants using manipulated carotenoid biosynthesis. PMID: 11164372 [PubMed - indexed for MEDLINE] 2051: Science. 2001 Jan 19;291(5503):490-3. Entrainment of the circadian clock in the liver by feeding. Stokkan KA, Yamazaki S, Tei H, Sakaki Y, Menaker M. National Science Foundation Center for Biological Timing and Department of Biology, University of Virginia, P.O. Box 400328, Charlottesville, VA 22904-4328, USA. mm7e@virginia.edu Circadian rhythms of behavior are driven by oscillators in the brain that are coupled to the environmental light cycle. Circadian rhythms of gene expression occur widely in peripheral organs. It is unclear how these multiple rhythms are coupled together to form a coherent system. To study such coupling, we investigated the effects of cycles of food availability (which exert powerful entraining effects on behavior) on the rhythms of gene expression in the liver, lung, and suprachiasmatic nucleus (SCN). We used a transgenic rat model whose tissues express luciferase in vitro. Although rhythmicity in the SCN remained phase-locked to the light-dark cycle, restricted feeding rapidly entrained the liver, shifting its rhythm by 10 hours within 2 days. Our results demonstrate that feeding cycles can entrain the liver independently of the SCN and the light cycle, and they suggest the need to reexamine the mammalian circadian hierarchy. They also raise the possibility that peripheral circadian oscillators like those in the liver may be coupled to the SCN primarily through rhythmic behavior, such as feeding. Publication Types: Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. PMID: 11161204 [PubMed - indexed for MEDLINE] 2052: Lancet. 2000 Mar 11;355(9207):931. Comment on: Lancet. 2000 Jan 29;355(9201):414. Toxins and genetically modified food. Trewavas A. Publication Types: Comment Letter PMID: 10752731 [PubMed - indexed for MEDLINE] 2053: J Agric Food Chem. 2000 Dec;48(12):5924-8. Fast apple (Malus x domestica) and tobacco (Nicotiana tobacum) leaf polyphenol oxidase activity assay for screening transgenic plants. Broothaerts W, McPherson J, Li B, Randall E, Lane WD, Wiersma PA. Pacific Agri-Food Research Centre, Agriculture and Agri-Food Canada, Summerland, British Columbia V0H 1Z0, Canada. wim.broothaerts@agr.kuleuven.ac.be A spectrophotometric assay method for the analysis of polyphenol oxidase (PPO), in apple and tobacco leaves, has been optimized to increase efficiency in the screening of large numbers of transgenic plants. Crude protein extracts from leaf punches were prepared in a FastPrep homogenizer. The addition of Triton X-100 during extraction resulted in 44 and 74% increases in the PPO activity recovered, from apple and tobacco, respectively. The enzyme kinetics differed markedly between apple and tobacco. Apple leaf PPO was isolated in a latent state and was activated by the addition of SDS. In contrast, tobacco PPO activity was inhibited by SDS, particularly at acidic pH. Apple PPO showed a pronounced pH optimum around pH 6, whereas the pH profile for tobacco PPO was much flatter, with a broad optimum around pH 4. The calculated Km' value for apple PPO, using 4-methylcatechol as substrate, was 8.1, and for tobacco the Km was 4.3. The PPO reaction was strongly inhibited by tropolone, a Cu competitor, and restored by the addition of Cu2+. Several factors affecting variability in leaf PPO activity levels in plants are discussed. PMID: 11141262 [PubMed - indexed for MEDLINE] 2054: Berl Munch Tierarztl Wochenschr. 2000 Nov-Dec;113(11-12):454-8. [Detection of genetically modified soy (Roundup-Ready) in processed food products] [Article in German] Hagen M, Beneke B. Staatliches Veterinäruntersuchungsamt Detmold. In this study, the application of a qualitative and a quantitative method of analysis to detect genetically modified RR-Soy (Roundup-Ready Soy) in processed foods is described. A total of 179 various products containing soy such as baby food and diet products, soy drinks and desserts, tofu and tofu products, soy based meat substitutes, soy protein, breads, flour, granules, cereals, noodles, soy bean sprouts, fats and oils as well as condiments were investigated following the pattern of the section 35 LMBG-method L 23.01.22-1. The DNA was extracted from the samples and analysed using a soybean specific lectin gene PCR as well as a PCR, specific for the genetic modification. Additional, by means of PCR in combination with fluorescence-detection (TaqMan 5'-Nuclease Assay), suspicious samples were subjected to a real-time quantification of the percentage of genetically modified RR-Soy. The methods of analysis proved to be extremely sensitive and specific in regard to the food groups checked. The fats and oils, as well as the condiments were the exceptions in which amplifiable soy DNA could not be detected. The genetic modification of RR-Soy was detected in 34 samples. Eight of these samples contained more than 1% of RR-Soy. It is necessary to determine the percentage of transgenic soy in order to assess whether genetically modified ingredients were deliberately added, or whether they were caused by technically unavoidable contamination (for example during transportation and processing). Publication Types: English Abstract PMID: 11153227 [PubMed - indexed for MEDLINE] 2055: Newsweek. 2000 Dec;136(26A):56-8. DNA on the dinner table. Cowley G. Publication Types: News PMID: 11147308 [PubMed - indexed for MEDLINE] 2056: Nat Biotechnol. 2001 Jan;19(1):15. GM vines: focal point for EC power struggle. Meldolesi A. Publication Types: News PMID: 11135541 [PubMed - indexed for MEDLINE] 2057: Nat Biotechnol. 2001 Jan;19(1):3. GM crop data-agronomy and ecology in tandem. Stewart CN Jr, Wheaton SK. Department of Biology, University of North Carolina-Greensboro, Greensboro, NC 27402, USA. nstewart@uncg.edu PMID: 11135511 [PubMed - indexed for MEDLINE] 2058: Med J Malaysia. 2000 Aug;55 Suppl B:62-4. Ethical issues of genetically modified foods (GMF). Shamsinar BA. Food Quality Control Division, Ministry of Health, 4th Floor, Block E, Komplek Pejabat-Pejabat, Jalan Dungun, 50490 Kuala Lumpur. Publication Types: Evaluation Studies PMID: 11125525 [PubMed - indexed for MEDLINE] 2059: Am J Clin Nutr. 2001 Jan;73(1):80-5. Absorption of iron from unmodified maize and genetically altered, low-phytate maize fortified with ferrous sulfate or sodium iron EDTA. Mendoza C, Viteri FE, Lönnerdal B, Raboy V, Young KA, Brown KH. Institute of Nutrition of Central America and Panama, Guatemala City, Guatemala. cmendoza@ucdavis.edu BACKGROUND: Reducing the phytate content in grains by genetic manipulation is a novel approach to increasing nonheme-iron absorption from mixed diets. Fractional iron absorption from a genetically modified strain of low-phytate maize (LPM) increased significantly, by 50%. OBJECTIVE: We assessed iron absorption from porridges prepared from the same LPM (lpa-1-1 mutant) and unmodified wild-type maize (WTM), both of which were fortified with either ferrous sulfate or sodium iron EDTA. DESIGN: Porridges providing 3.4 mg Fe were fortified with either ferrous sulfate or sodium iron EDTA to provide an additional 1 mg Fe/serving. In 14 nonanemic women, iron absorption was measured as the amount of radioiron incorporated into red blood cells (extrinsic tag method) 12 d after consumption of the study diets. RESULTS: No significant effect of phytate content on iron absorption was found when porridge was fortified with either sodium iron EDTA or ferrous sulfate. Fractional absorption of iron from WTM porridge fortified with sodium iron EDTA (5.73%) was 3.39 times greater than that from the same porridge fortified with ferrous sulfate (1.69%). Fractional absorption of iron from the sodium iron EDTA-fortified LPM porridge (5.40%) was 2.82 times greater than that from LPM porridge fortified with ferrous sulfate (1.91%) (P<0.0001 for both comparisons, repeated-measures analysis of variance). Thus, the previously identified benefit of LPM was no longer detectable when maize porridge was fortified with additional iron. CONCLUSION: Iron was absorbed more efficiently when the fortificant was sodium iron EDTA rather than ferrous sulfate, regardless of the type of maize. Publication Types: Research Support, Non-U.S. Gov't PMID: 11124754 [PubMed - indexed for MEDLINE] 2060: Int Microbiol. 2000 Mar;3(1):1-2. Genetically modified foods: a case of information or misinformation? Ramón D. Publication Types: Editorial PMID: 10963326 [PubMed - indexed for MEDLINE] 2061: Prog Lipid Res. 2000 Nov;39(6):477-506. Analysis of genetically modified oils. Hazebroek JP. Trait and Technology Development, Analytical/Biochemistry Department, Pioneer Hi-Bred International, Inc., PO Box 1004, 7300 NW 62nd Avenue, 50131, Johnston, IA, USA. Genetically modified oils with altered functional or nutritional characteristics are being introduced into the marketplace. A wide array of analytical techniques has been utilized to facilitate developing these oils. This article attempts to review the utilization of these analytical procedures for characterizing both the chemistry and some functionality of these oils. Although techniques to assess oxidative stability in frying and food applications are covered, measurement of nutritional characteristics are not. Publication Types: Review PMID: 11106811 [PubMed - indexed for MEDLINE] 2062: J Am Diet Assoc. 2000 Nov;100(11):1308-11. Solving global nutrition challenges requires more than new biotechnologies. Babcock BC, Francis CA. Department of Agronomy, University of Nebraska, Lincoln, USA. bcbabcock@navix.net PMID: 11103651 [PubMed - indexed for MEDLINE] 2063: J Am Diet Assoc. 2000 Nov;100(11):1306-8. Comment in: J Am Diet Assoc. 2001 Mar;101(3):289-90. Biotechnology: mobilizing dietitians to be a resource. Greger JL. Department of Nutritional Sciences, University of Wisconsin-Madison 53706, USA. PMID: 11103650 [PubMed - indexed for MEDLINE] 2064: Curr Opin Immunol. 2000 Dec;12(6):647-53. Mechanisms of food allergy. Helm RM, Burks AW. University of Arkansas for Medical Sciences, Arkansas Children's Hospital Research Institute, 1120 Marshall Street, Little Rock, AR 72202, USA. helmrickim@exchange.uams.edu The prevalence of food allergy continues to rise, particularly in 'westernized' societies; it has been linked to the 'hygiene hypothesis' and the increased diversity of food consumption worldwide. The pathogenic mechanisms and Th1/Th2 paradigm are being closely examined with respect to the occurrence of inflammatory and injury/repair responses at different mucosal sites. Genetically modified plants as potential food sources and allergenicity are current topics of controversy. Publication Types: Review PMID: 11102767 [PubMed - indexed for MEDLINE] 2065: J Agric Food Chem. 2000 Nov;48(11):5243-8. Transgenic apple (Malus x domestica) shoot showing low browning potential. Murata M, Haruta M, Murai N, Tanikawa N, Nishimura M, Homma S, Itoh Y. Department of Nutrition and Food Science, Ochanomizu University, 2-1-1 Otsuka, Bunkyo-ku, Tokyo 112-8610, Japan. murata@cc.ocha.ac.jp Transgenic apple shoots were prepared from leaf disks by using Agrobacterium tumefaciens carrying the kanamycin (KM) resistance gene and antisense polyphenol oxidase (PPO) DNA. Four transgenic apple lines that grew on the medium containing 50 microgram/mL KM were obtained. They contained the KM resistance gene and grew stably on the medium for >3 years. Two transgenic shoot lines containing antisense PPO DNA in which PPO activity was repressed showed a lower browning potential than a control shoot. Publication Types: Research Support, Non-U.S. Gov't PMID: 11087467 [PubMed - indexed for MEDLINE] 2066: EMBO J. 2000 Nov 15;19(22):6150-61. Transcriptional repression by AtMYB4 controls production of UV-protecting sunscreens in Arabidopsis. Jin H, Cominelli E, Bailey P, Parr A, Mehrtens F, Jones J, Tonelli C, Weisshaar B, Martin C. John Innes Centre, Colney, Norwich NR4 7UH, Institute of Food Research, Colney, Norwich NR4 7UH, UK. An Arabidopsis thaliana line that is mutant for the R2R3 MYB gene, AtMYB4, shows enhanced levels of sinapate esters in its leaves. The mutant line is more tolerant of UV-B irradiation than wild type. The increase in sinapate ester accumulation in the mutant is associated with an enhanced expression of the gene encoding cinnamate 4-hydroxylase, which appears to be the principal target of AtMYB4 and an effective rate limiting step in the synthesis of sinapate ester sunscreens. AtMYB4 expression is downregulated by exposure to UV-B light, indicating that derepression is an important mechanism for acclimation to UV-B in A.thaliana. The response of target genes to AtMYB4 repression is dose dependent, a feature that operates under physiological conditions to reinforce the silencing effect of AtMYB4 at high activity. AtMYB4 works as a repressor of target gene expression and includes a repression domain. It belongs to a novel group of plant R2R3 MYB proteins involved in transcriptional silencing. The balance between MYB activators and repressors on common target promoters may provide extra flexibility in transcriptional control. Publication Types: Research Support, Non-U.S. Gov't PMID: 11080161 [PubMed - indexed for MEDLINE] 2067: Nat Biotechnol. 2000 Nov;18(11):1137-8. Italian GMO ban could spread. Meldolesi A. Publication Types: News PMID: 11062425 [PubMed - indexed for MEDLINE] 2068: Nat Biotechnol. 2000 Nov;18(11):1136-7. Taco dispute underscores need for standardized tests. Dorey E. Publication Types: News PMID: 11062423 [PubMed - indexed for MEDLINE] 2069: Pediatr Int. 2000 Oct;42(5):461-3. Child health in the new millennium. Gracey M. School of Public Health, Curtin University of Technology, Perth, WA, Australia. In today's modern, industrialized and affluent countries, like Japan and Australia, better living conditions and hygiene, plentiful nutritious food and rapid advances in biology and medical technologies have helped to bring about dramatic improvements in child health. The previous heavy burdens of infections and undernutrition have been eliminated or can now be controlled or effectively treated. In these countries, child health standards are higher than ever and expectation of life at birth is much higher than in the past. Some of the technological advances that have helped bring about this transformation are immunization, antimicrobial therapy, successful treatment of childhood leukemias, transplantation of vital organs and implementation of genetic diagnosis and gene therapy. The use of genetically modified foods and the prospects for cloning of humans are areas of intense interest and controversy. However, these advances have their disadvantages (e.g. antibiotic-induced drug resistance). Urbanization has encouraged the 'westernization' of dietary patterns and the long-term 'lifestyle diseases' that can follow in adults. Accidents, violence and drug abuse are major problems in many parts of the world. Changes in attitudes to sexuality and the spread of HIV/AIDS is another major problem, especially in Africa and Asia. Environmental pollution and the degradation of agricultural lands, rivers and seas are also important. Ironically, standards of child health and the prospects for long life in countries like Japan are better than ever before, but social and environmental changes are presenting children and their carers with new and unanswered challenges as we enter the 21 st century and the new millennium. PMID: 11059531 [PubMed - indexed for MEDLINE] 2070: Biotechniques. 2000 Oct;29(4):832-6, 838-43. Transgenic plants and biosafety: science, misconceptions and public perceptions. Stewart CN Jr, Richards HA 4th, Halfhill MD. Department of Biology, University of North Carolina, Greensboro 27402-6174, USA. nstewart@uncg.edu One usually thinks of plant biology as a non-controversial topic, but the concerns raised over the biosafety of genetically modified (GM) plants have reached disproportionate levels relative to the actual risks. While the technology of changing the genome of plants has been gradually refined and increasingly implemented, the commercialization of GM crops has exploded. Today's commercialized transgenic plants have been produced using Agrobacterium tumefaciens-mediated transformation or gene gun-mediated transformation. Recently, incremental improvements of biotechnologies, such as the use of green fluorescent protein (GFP) as a selectable marker, have been developed. Non-transformation genetic modification technologies such as chimeraplasty will be increasingly used to more precisely modify germplasm. In spite of the increasing knowledge about genetic modification of plants, concerns over ecological and food biosafety have escalated beyond scientific rationality. While several risks associated with GM crops and foods have been identified, the popular press, spurred by colorful protest groups, has left the general public with a sense of imminent danger. Reviewed here are the risks that are currently under research. Ecological biosafety research has identified potential risks associated with certain crop/transgene combinations, such as intra- and interspecific transgene flow, persistence and the consequences of transgenes in unintended hosts. Resistance management strategies for insect resistance transgenes and non-target effects of these genes have also been studied. Food biosafety research has focused on transgenic product toxicity and allergenicity. However, an estimated 3.5 x 10(12) transgenic plants have been grown in the U.S. in the past 12 years, with over two trillion being grown in 1999 and 2000 alone. These large numbers and the absence of any negative reports of compromised biosafety indicate that genetic modification by biotechnology poses no immediate or significant risks and that resulting food products from GM crops are as safe as foods from conventional varieties. We are increasingly convinced that scientists have a duty to conduct objective research and to effectively communicate the results--especially those pertaining to the relative risks and potential benefits--to scientists first and then to the public. All stakeholders in the technology need more effective dialogues to better understand risks and benefits of adopting or not adopting agricultural biotechnologies. Publication Types: Research Support, U.S. Gov't, Non-P.H.S. PMID: 11056815 [PubMed - indexed for MEDLINE] 2071: Biosci Biotechnol Biochem. 2000 Sep;64(9):1881-6. Safety assessment of rice genetically modified with soybean glycinin by feeding studies on rats. Momma K, Hashimoto W, Yoon HJ, Ozawa S, Fukuda Y, Kawai S, Takaiwa F, Utsumi S, Murata K. Research Institute for Food Science, Kyoto University, Uji, Japan. momma@food2.food.kyoto-u.ac.jp Feeding studies on rice genetically modified with soybean glycinin were performed on rats for four weeks. The rats were divided into three groups, each being fed on (I) only a commercial diet, (II) this diet plus control rice and (III) this diet plus rice genetically modified with glycinin. The rats were fed with 10 g/kg-weight of rice every day by oral administration. During the test period, the rats in every group grew well without marked differences in appearance, food intake, body weight, or cumulative body weight gain. There were also no significant differences in the blood count, blood composition or internal organ weights among the rats. Necropsy at the end of the experiment indicated neither pathological symptoms nor histopathological abnormalities in the liver and kidney. Judging from these results, the rice genetically modified with glycinin is considered to have been essentially the same in nutritional and biochemical characteristics as the control rice. Publication Types: Research Support, Non-U.S. Gov't PMID: 11055391 [PubMed - indexed for MEDLINE] 2072: Nature. 2000 Oct 12;407(6805):765-7. Messenger RNA targeting of rice seed storage proteins to specific ER subdomains. Choi SB, Wang C, Muench DG, Ozawa K, Franceschi VR, Wu Y, Okita TW. Institute of Biological Chemistry, Washington State University, Pullman 99164, USA. Rice seeds, a rich reserve of starch and protein, are a major food source in many countries. Unlike the seeds of other plants, which typically accumulate one major type of storage protein, rice seeds use two major classes, prolamines and globulin-like glutelins. Both storage proteins are synthesized on the endoplasmic reticulum (ER) and translocated to the ER lumen, but are then sorted into separate intracellular compartments. Prolamines are retained in the ER lumen as protein bodies whereas glutelins are transported and stored in protein storage vacuoles. Mechanisms responsible for the retention of prolamines within the ER lumen and their assembly into intracisternal inclusion granules are unknown, but the involvement of RNA localization has been suggested. Here we show that the storage protein RNAs are localized to distinct ER membranes and that prolamine RNAs are targeted to the prolamine protein bodies by a mechanism based on RNA signal(s), a process that also requires a translation initiation codon. Our results indicate that the ER may be composed of subdomains that specialize in the synthesis of proteins directed to different compartments of the plant endomembrane system. Publication Types: Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. PMID: 11048726 [PubMed - indexed for MEDLINE] 2073: J Allergy Clin Immunol. 2000 Oct;106(4):752-62. Digestibility of allergens extracted from natural rubber latex and vegetable foods. Yagami T, Haishima Y, Nakamura A, Osuna H, Ikezawa Z. Division of Medical Devices, National Institute of Health Sciences, Tokyo, Japan. BACKGROUND: Several cross-reactive allergens are now known to be involved in the defense responses of higher plants. Such proteins are drawing the attention of plant breeders because of their antimicrobial or stress-alleviating activities. Plants genetically modified to express defense-related proteins are being developed. The current concern is focused on the allergenicity of these intentionally expressed proteins. OBJECTIVE: It is believed that food allergens are proteins resistant to digestion. Digestibility tests have been accepted as an appropriate method for evaluating the allergenicity of newly introduced proteins. In this study we investigated the usefulness of this method for detecting allergens from natural rubber latex and vegetable foods. METHODS: Proteins were extracted from rubber latex, potato, and 5 kinds of fruits. Simulated gastric fluid (SGF) and simulated intestinal fluid (SIF) were used for the digestibility test. An aliquot of each digest was periodically withdrawn and analyzed. Allergens were detected with pooled sera from individuals with latex allergy or patients given a diagnosis of oral allergy syndrome. RESULTS: Most latex and vegetable food proteins were digested by the SGF within 4 minutes. Numerous allergens were also decomposed by the SGF within 8 minutes. Although vegetable food allergens were relatively stable in the SIF, kiwi allergens were substantially degraded by the SIF within 16 hours. CONCLUSION: The pronounced lability of the plant-derived allergens was thought to reflect the discrete sensitization and elicitation processes of patients with latex-fruit syndrome or oral allergy syndrome. These results indicate that the allergenicity of a newly expressed protein should be carefully evaluated according to not only its digestibility but also other important properties. PMID: 11031347 [PubMed - indexed for MEDLINE] 2074: Plant Physiol. 2000 Oct;124(2):487-90. Ending world hunger. The promise of biotechnology and the threat of antiscience zealotry. Borlaug NE. Publication Types: Editorial PMID: 11027697 [PubMed - indexed for MEDLINE] 2075: Curr Opin Biotechnol. 2000 Oct;11(5):505-8. Gene transfer from genetically modified food. Gasson MJ. Food Safety Science Division, Institute of Food Research, Norwich Research Park, Colney, Norfolk, NR4 7UA, Norwich, UK. mike.gasson@bbsrc.ac.uk The current debate about the safety of genetically modified food includes some important scientific issues where more scientific data would aid the robustness of safety evaluation. One example is the possibility of gene transfer, especially from genetically modified plant material. Publication Types: Review PMID: 11024371 [PubMed - indexed for MEDLINE] 2076: Eur J Endocrinol. 2000 Oct;143(4):535-41. Development of obesity in transgenic rats with low circulating growth hormone levels: involvement of leptin resistance. Furuhata Y, Kagaya R, Hirabayashi K, Ikeda A, Chang KT, Nishihara M, Takahashi M. Department of Veterinary Physiology, Veterinary Medical Science, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan. BACKGROUND: Human growth hormone (hGH) transgenic (TG) rats have been produced in our laboratory. These TG rats are characterized by low circulating hGH levels, virtually no endogenous rGH secretion, and massive obesity. OBJECTIVE: To elucidate how energy balance and leptin sensitivity contributed to the establishment of this obesity. DESIGN AND METHODS: Food intake, locomotor activity and leptin concentrations in serum and cerebrospinal fluid were measured in TG rats and their non-transgenic littermates (control). The effect of intraperitoneal and intracerebroventricular injection of leptin on food intake and body weight gain was also examined. RESULTS: An increase in food intake and a decrease in locomotor activity were observed from 4 and 7 weeks of age, respectively, in the transgenic rats compared with control. Serum leptin concentrations of the transgenic rats were more than twice as high as those of control rats and were associated with an increased white adipose tissue mass and ob gene expression. Intraperitoneal injection of leptin significantly decreased food intake and body weight gain in control rats, but not in transgenic rats. Leptin concentration in the cerebrospinal fluid of transgenic rats was not different from that of control rats, and intracerebroventricular injection of leptin was similarly effective in reducing food intake and body weight gain as it was in control rats. CONCLUSIONS: These results suggest that the transgenic rats, whose GH secretion is suppressed, develop obesity due to early onset of an increase in food intake and a decrease in locomotor activity with leptin resistance resulting from deteriorating leptin transport from peripheral blood to cerebrospinal fluid. Publication Types: Research Support, Non-U.S. Gov't PMID: 11022201 [PubMed - indexed for MEDLINE] 2077: Nat Biotechnol. 2000 Oct;18(10):1018-9. Uninformation and the choice paradox. McHughen A. University of Saskatchewan, Saskatoon, SK S7N 5A8, Canada mchughen@duke.usask.ca PMID: 11016993 [PubMed - indexed for MEDLINE] 2078: Endocrinology. 2000 Oct;141(10):3912-5. Estrogen agonist and antagonist action on the human estrogen receptor in Drosophila. Thackray VG, Young RH, Hooper JE, Nordeen SK. Program in Molecular Biology, University of Colorado Health Sciences Center, Denver 80262, USA. The estrogen receptor (ER) regulates the expression of genes involved in the growth, proliferation and differentiation of skeletal, cardiovascular, neural and reproductive tissues. A basic scheme for the mechanism for ER action has been developed, but precise details on the interactions between ER and the cellular signaling and transcription machinery required for receptor-mediated regulation of specific target genes are still lacking. We have developed a genetic approach to explore the functional interactions of ER. In this work, we describe the development of an estrogen responsive system in the fruit fly, Drosophila melanogaster. Transgenic flies carrying the human ER alpha and an estrogen responsive green fluorescent protein (GFP) reporter gene were constructed. In vivo expression of the GFP reporter gene was observed when larvae were grown on a food source containing steroidal or nonsteroidal estrogens. The induction of the reporter gene by estrogens was blocked upon treatment with tamoxifen, an estrogen antagonist. However, we failed to recapitulate ligand-independent activation of the receptor in vivo or in cultured Drosophila cells. An estrogen responsive Drosophila system could be used to identify and characterize the complex functional interactions between ER and the other components of the cellular transcriptional apparatus. Publication Types: Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. PMID: 11014249 [PubMed - indexed for MEDLINE] 2079: EMBO J. 2000 Oct 2;19(19):5148-56. A Caenorhabditis elegans MAP kinase kinase, MEK-1, is involved in stress responses. Koga M, Zwaal R, Guan KL, Avery L, Ohshima Y. Department of Biology, Faculty of Sciences, Kyushu University Graduate School, Fukuoka 812-8581, Japan. The c-Jun N-terminal kinase (JNK), a member of the mitogen-activated protein kinase (MAPK) family, was shown to be involved in the response to various stresses in cultured cells. However, there is little in vivo evidence indicating a role for a JNK pathway in the stress response of an organism. We identified the Caenorhabditis elegans mek-1 gene, which encodes a 347 amino acid protein highly homologous to mammalian MKK7, an activator of JNK. Mek-1 reporter fusion proteins are expressed in pharyngeal muscle, uterus, a portion of intestine, and neurons. A mek-1 deletion mutant is hypersensitive to copper and cadmium ions and to starvation. A wild-type mek-1 transgene rescued the hypersensitivity to the metal ions. Double mutants of mek-1 with an eat-5, eat-11 or eat-18 mutation, which are characterized by a limited feeding defect, showed distinct growth defects under normal conditions. Expression of an activated form of MEK-1 in the whole animal or specifically in the pharynx inhibited pharyngeal pumping. These results suggest a role for mek-1 in stress responses, with a focus in the pharynx and/or intestine. Publication Types: Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. PMID: 11013217 [PubMed - indexed for MEDLINE] 2080: Behav Brain Res. 2000 Oct;115(1):1-8. Temporal and spatial adaptation to food restriction in mice under naturalistic conditions. Dell'Omo G, Ricceri L, Wolfer DP, Poletaeva II, Lipp H. Institute of Anatomy and Center for Neuroscience, University of Zürich-Irchel, Winterhurerstrasse 190, CH-8057, Zurich, Switzerland. Free-living female laboratory mice, adapted to outdoor life in large pens providing a naturalistic environment, were tested for their ability to modify their foraging habits to controlled food supply. An automatic feeder box delivered a small portion of the daily quantity of seeds to each individual mouse. Eight such boxes were placed into an outdoor pen. Each day, mice had to visit all boxes to gather the daily amount of food and were rewarded only at the first visit to each box. Mice were individually recognised by an implanted microchip. Throughout a 16-day period, feeding activity concentrated in an interval time around the beginning of the daily session. During the same period, the number of different feeders visited every day by mice increased irrespective of variation in exploratory activity. The experimental set-up allowed detecting temporal and spatial adaptations to the food restriction, as well as behavioural differences due to territorial and social factors. These data permit the design of novel tests assessing behavioural changes, memory and learning in normal and genetically modified mice, both in the laboratory and in naturalistic settings. Publication Types: Research Support, Non-U.S. Gov't PMID: 10996402 [PubMed - indexed for MEDLINE] 2081: FEBS Lett. 2000 Sep 15;481(2):164-8. DNA stability in plant tissues: implications for the possible transfer of genes from genetically modified food. Chiter A, Forbes JM, Blair GE. School of Biochemistry and Molecular Biology, Room 8.10a Garstang Building, University of Leeds, Mount Preston Street, Leeds, UK. The potential for transfer of antibiotic resistance genes from genetically modified (GM) plant material to microbes through genetic recombination in the human or animal gut is a consideration that has engendered caution in the use of GM foods. This study was aimed at defining the optimal physical and chemical conditions necessary to ensure sufficient fragmentation of DNA in plant tissues to a size where it would be unlikely to be stably transferred to bacterial gut microflora. The ribulose 1,5-bisphosphate carboxylase/oxygenase small subunit (Rubisco SS) genes are of similar size (approximately 1.4 kb) to transgenes present in GM plants. DNA analysis and PCR amplification of Rubisco SS genes showed that fresh maize and maize silage contained high molecular weight DNA and intact Rubisco SS genes. Relatively high temperatures and pressurised steam were necessary to degrade fully genomic DNA and Rubisco SS genes in maize and wheat grains, the source of most animal feedstuffs. Furthermore, chemical expulsion and extrusion of oilseeds resulted in residues with completely degraded genomic DNA. These results imply that stringent conditions are needed in the processing of GM plant tissues for feedstuffs to eliminate the possibility of transmission of transgenes. Publication Types: Research Support, Non-U.S. Gov't PMID: 10996317 [PubMed - indexed for MEDLINE] 2082: J AOAC Int. 2000 Jul-Aug;83(4):919-27. Validation of an immunoassay for detection and quantitation of a genetically modified soybean in food and food fractions using reference materials: interlaboratory study. Lipp M, Anklam E, Stave JW, Lipp M, Anklam E, Stave JW. European Commission, Joint Research Center, Institute for Health and Consumer Protection, Food Products and Consumer Goods Unit, Ispra (Va), Italy. An immunoassay for detection of a specific genetically modified soybean (Roundup-Ready) was validated on dried soybean powder in an interlaboratory study. Different percentages of genetically modified soybeans in nonmodified soybean matrix were evaluated in a blind study. Thirty-eight laboratories from 13 countries participated. The immunoassay was evaluated for 2 endpoints: (1) To give a semiquantitative result, i.e., determination of a given sample above or below a given threshold, or (2) to compute a quantitative result, i.e., percentage of genetically modified soybeans in the sample. Semiquantitative results showed that a given sample which contained <2% genetically modified soybeans was identified as below 2% with a 99% confidence level. Quantitative use of the assay resulted in a repeatability (r) and reproducibility (R) that were computed to be RSDr = 7% and RSDR = 10%, respectively, for a sample containing 2% genetically modified soybeans. Application of this method depends on availability of appropriate reference materials for a specific food matrix. Only matrix-matched reference materials can be used for analysis of food or food fractions. PMID: 10995116 [PubMed - indexed for MEDLINE] 2083: Science. 2000 Aug 25;289(5483):1279-81. Comment in: Science. 2000 Nov 24;290(5496):1505-6. Agrobiotechnology. Asia gets a taste of genetic food fights. Normile D. Despite the need to feed growing populations in Asia, controversy over genetically modified foods is putting a damper on efforts to develop and test new crops. The signs of opposition are scattered and vary from country to country. But they are enough to be worrisome to the region's leaders. Publication Types: News PMID: 10979850 [PubMed - indexed for MEDLINE] 2084: Med J Aust. 2000 Aug 7;173(3):167. Comment on: Med J Aust. 2000 Feb 21;172(4):170-3. Genetically modified foods--safety and regulatory issues. Liehne PF. Publication Types: Comment Letter PMID: 10979390 [PubMed - indexed for MEDLINE] 2085: Med J Aust. 2000 Aug 7;173(3):166-7. Comment on: Med J Aust. 2000 Feb 21;172(4):170-3. Genetically modified foods--safety and regulatory issues. Clinch-Jones CA. Publication Types: Comment Letter PMID: 10979389 [PubMed - indexed for MEDLINE] 2086: Sci Am. 2000 Sep;283(3):66-71. Edible vaccines. Langridge WH. Publication Types: Review PMID: 10976468 [PubMed - indexed for MEDLINE] 2087: Mol Plant Microbe Interact. 2000 Sep;13(9):962-74. Cell-to-cell movement of potexviruses: evidence for a ribonucleoprotein complex involving the coat protein and first triple gene block protein. Lough TJ, Netzler NE, Emerson SJ, Sutherland P, Carr F, Beck DL, Lucas WJ, Forster RL. Horticulture and Food Research Institute of New Zealand, Plant Health and Development Group, Auckland. tlough@hort.cri.nz The triple gene block proteins (TGBp1-3) and coat protein (CP) of potexviruses are required for cell-to-cell movement. Separate models have been proposed for intercellular movement of two of these viruses, transport of intact virions, or a ribonucleoprotein complex (RNP) comprising genomic RNA, TGBp1, and the CP. At issue therefore, is the form(s) in which RNA transport occurs and the roles of TGBp1-3 and the CP in movement. Evidence is presented that, based on microprojectile bombardment studies, TGBp1 and the CP, but not TGBp2 or TGBp3, are co-translocated between cells with viral RNA. In addition, cell-to-cell movement and encapsidation functions of the CP were shown to be separable, and the rate-limiting factor of potexvirus movement was shown not to be virion accumulation, but rather, the presence of TGBp1-3 and the CP in the infected cell. These findings are consistent with a common mode of transport for potexviruses, involving a non-virion RNP, and show that TGBp1 is the movement protein, whereas TGBp2 and TGBp3 are either involved in intracellular transport or interact with the cellular machinery/docking sites at the plasmodesmata. Publication Types: Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. PMID: 10975653 [PubMed - indexed for MEDLINE] 2088: Nat Biotechnol. 2000 Sep;18(9):920. Withdrawn Greenpeace Bt suit enters spin cycle. Fox JL. Publication Types: News PMID: 10973201 [PubMed - indexed for MEDLINE] 2089: Nat Biotechnol. 2000 Sep;18(9):919-20. Comment in: Nat Biotechnol. 2000 Dec;18(12):1227. Green ag minister wreaks havoc on Italy's agbiotech. Meldolesi A. Publication Types: News PMID: 10973200 [PubMed - indexed for MEDLINE] 2090: Nat Biotechnol. 2000 Sep;18(9):918-9. National politicians block GM progress. Hodgson J. Publication Types: News PMID: 10973199 [PubMed - indexed for MEDLINE] 2091: Arch Latinoam Nutr. 1999 Sep;49(3 Suppl 2):47S-51S. [New alternatives in the prevention of iron deficiency. Use of genetic engineering in food modification] [Article in Spanish] García-Casal MN. Centro de Medicina Experimental, Instituto Venezolano de Investigaciones Científicas (IVIC), Caracas. This article reviews the possible applications of new food biotechnology techniques to introduce some compounds into plants or animals. The potential for these plant modification methods has ample applications ranging from improvements in food production and development for human consumption, production of antibodies or therapeutic proteins, inclusion of nutrients to improve nutritional value of the food to production of vaccines. It must be clear though that currently the scope and consequences of such modifications are not completely clear. There is some concern about potential secondary effects and the hypothesis of the appearance of new viruses due to recombinant genetical transformations that have not been totally rejected. However the tendency is towards considering the process as safe. Finally some evidence is presented about the possibility of introducing the capacity to synthesize vitamin A in vegetables or produce rice with high content of iron as real alternatives to fight some of the nutritional deficiencies most common worldwide. Publication Types: English Abstract PMID: 10971836 [PubMed - indexed for MEDLINE] 2092: Science. 2000 Sep 1;289(5484):1554-7. Comment in: Science. 2000 Sep 1;289(5484):1481-2. Predictions of biodiversity response to genetically modified herbicide-tolerant crops. Watkinson AR, Freckleton RP, Robinson RA, Sutherland WJ. Schools of Environmental and Biological Sciences, University of East Anglia, Norwich NR4 7TJ, UK. a.watkinson@uea.ac.uk We simulated the effects of the introduction of genetically modified herbicide-tolerant (GMHT) crops on weed populations and the consequences for seed-eating birds. We predict that weed populations might be reduced to low levels or practically eradicated, depending on the exact form of management. Consequent effects on the local use of fields by birds might be severe, because such reductions represent a major loss of food resources. The regional impacts of GMHT crops are shown to depend on whether the adoption of GMHT crops by farmers covaries with current weed levels. Publication Types: Research Support, Non-U.S. Gov't PMID: 10968791 [PubMed - indexed for MEDLINE] 2093: Appl Microbiol Biotechnol. 2000 Aug;54(2):180-5. A genetically modified solvent-tolerant bacterium for optimized production of a toxic fine chemical. Wery J, Mendes da Silva DI, de Bont JA. Department of Food Technology and Nutritional Sciences, Wageningen University, The Netherlands. Jan.Wery@imb.ftns.wau.nl The aim of the study was to investigate whether toxic fine chemical production can be improved using the solvent-tolerant Pseudomonas putida S12 in a two-liquid-phase system consisting of aqueous media and a water-immiscible octanol phase with production of 3-methylcatechol from toluene as the model conversion. For this purpose the genes involved in this conversion, todC1C2BAD from P. putida F1, were introduced into P. putida S12 with high stable expression. Production of 3-methylcatechol was monitored in batch incubations with different media using a single medium and a two-liquid medium-octanol system. The maximum concentration of 3-methylcatechol increased two-fold using the two-liquid medium-octanol system, irrespective of the selected medium. PMID: 10968630 [PubMed - indexed for MEDLINE] 2094: Int Arch Occup Environ Health. 2000 Jun;73 Suppl:S14-8. Safety evaluation of genetically modified foods. Martens MA. Monsanto Europe SA, Louvain-la-Neuve, Belgium. The concept of substantial equivalence has been accepted as the cornerstone of the health hazard assessment of genetically modified (GM) foods (OECD 1993). Substantial equivalence is the most practical approach to address the safety of foods or food components derived from GM crops and is based on comparison of the phenotypic and compositional characteristics of the parent crop and the GM crop. Basically, three categories of GM crops can be considered (FAO/WHO 1996; EU 1997): (a) GM crops which have the same composition as the parent crop, (b) GM crops which have the same composition as the parent crop with the exception of a well-defined trait, and (c) GM crops which are different from the parent crop. For the safety assessment of the first category of GM foods only a molecular characterisation of the genetic insert is sufficient, whereas for the second category a safety assessment of the expressed protein(s) is also required. For the last category an extensive evaluation including bioavailability and wholesomeness studies are required, beside the molecular characterisation and safety assessment of the expressed protein(s) and their products. By molecular characterisation is meant the position, nature, stability and number of copies of the inserted DNA. Substantial equivalence is established by the determination of the phenotypic characteristics (e.g. resistance against diseases, agronomic properties) and the complete chemical composition of the plant including nutrients, toxicants, antinutrients, and allergens. The toxicity of the expressed protein(s) is assessed by their homology with known protein toxins, degradation in the gastro-intestinal tract, stability to food processing and acute toxicity in rodents. The possible allergenicity of the expressed proteins is evaluated by comparison of their amino acid sequence with that of known allergens and determination of their stability to digestion and food processing. If the source of the genetic insert is allergenic then the use of solid-state immunoassays, skin prick tests and even food challenge tests can be considered. Publication Types: Review PMID: 10968554 [PubMed - indexed for MEDLINE] 2095: Nature. 2000 Aug 10;406(6796):560. Comment on: Nature. 2000 Apr 13;404(6779):689. Environment regulations hinder biotech industry. Miller HI. Publication Types: Comment Letter PMID: 10949276 [PubMed - indexed for MEDLINE] 2096: Rev Esp Salud Publica. 2000 May-Jun;74(3):255-61. [Health risks of genetically modified foods: a literature review] [Article in Spanish] Domingo Roig JL, Gómez Arnáiz M. Laboratorio de Toxicología y Salud Medioambiental, Facultad de Medicina, Universidad Rovira i Virgili, Tarragona. In 1999, there has been much concern on the safety of genetically modified (GM) foods, an important and complex area of safety research, which demands rigorous standards. Various groups, including consumers and environmental Non Governmental Organizations (NGO) have suggested that all GM foods should be subjected to long-term animal feeding studies before approval for human consumption. The main goal of this review has been to know which is the state-of-the art regarding to the potential adverse effects of GM foods. Two databases, MEDLINE and TOXLINE, as well as a number of URL from INTERNET were used for references. Although commentaries, general news and letters to the Editor have been frequently published in reputable journals, papers about experimental studies on the safety of GM foods are surprisingly very scant. If data on toxicological assessment of GM foods have been obtained, these have not been reported in scientific journals and subjected to the scientific judgement. Publication Types: English Abstract Review PMID: 10918812 [PubMed - indexed for MEDLINE] 2097: Rev Esp Salud Publica. 2000 May-Jun;74(3):211-4. [Genetically modified foods and current epidemiology] [Article in Spanish] Tormo Díaz MJ. Publication Types: Editorial PMID: 10918810 [PubMed - indexed for MEDLINE] 2098: Int J Epidemiol. 2000 Aug;29(4):730-3. Have lessons been learned from the UK bovine spongiform encephalopathy (BSE) epidemic? O'Brien M. History is full of examples of public health, commerce and politics in conflict. In recent years attempts to protect UK egg producers, after the discovery of Salmonella enteritidis (phage type 4) in hens' eggs, strained previously good working relationships between medical and veterinary epidemiologists and ended the political career of a government minister who spoke out in defence of the public health. Against the background lessons of earlier high profile public health problems in the UK conflict should have been avoided when bovine spongiform encephalopathy (BSE) started. It might have been expected that its significance for human health could have been recognized and researched earlier. Public announcements about it could have been timely and clear. Unfortunately this was not the case and it looks as though similar mistakes are going to be repeated over genetically modified foods. PMID: 10922352 [PubMed - indexed for MEDLINE] 2099: Public Health. 1999 May;113(3):103-4. Having your cake (genetically modified) and eating it. Beaver MW. Publication Types: Editorial PMID: 10910405 [PubMed - indexed for MEDLINE] 2100: Nature. 2000 Jul 13;406(6792):115. Scientists back GM for Third World. Dickson D. Publication Types: News PMID: 10910326 [PubMed - indexed for MEDLINE] 2101: Nutrition. 2000 Jul-Aug;16(7-8):709-11. Food risks from transgenic crops in perspective. Conner AJ, Jacobs JM. New Zealand Institute for Crop & Food Research Ltd., Christchurch. connert@crop.cri.nz Publication Types: Review PMID: 10906607 [PubMed - indexed for MEDLINE] 2102: Nutrition. 2000 Jul-Aug;16(7-8):706-9. Labeling of gene-spliced foods: a label we don't need. Miller HI. Hoover Institution, Stanford University, Stanford, CA 94305-6010, USA. miller@hoover.stanford.edu Publication Types: Review PMID: 10906606 [PubMed - indexed for MEDLINE] 2103: Anal Chem. 2000 Jul 1;72(13):454A-459A. Detecting genetically modified products in food. Erickson BE. PMID: 10905286 [PubMed - indexed for MEDLINE] 2104: Nat Biotechnol. 2000 Jul;18(7):705-6. EC struggles to unravel regulations. Hodgson J. Publication Types: News PMID: 10888826 [PubMed - indexed for MEDLINE] 2105: Br Med Bull. 2000;56(1):254-68. The future relationship between the media, the food industry and the consumer. Anderson WA. Food Safety Authority of Ireland, Dublin, Ireland. The relationship between the media, the food industry and the consumer is probably at its lowest point as we start the new millennium. The frequency of food scares appears to be increasing and news reports sometimes seem both sensational and polarised. High profile issues like the development of bovine spongiform encephalopathy in the UK and the dioxin contamination of poultry products in Belgium have undermined consumer confidence in the food industry. The recent genetically modified foods' debate has served to demonstrate the gulf that has grown between the food industry, food safety experts and the public. This is a rift that has been exploited by environmental pressure groups and fuelled by the media. This paper examines some of the underlying causes of the current air of mistrust that seems to exist between the media, the food industry and the consumer. Also, by examining the projected trends in these root causes, it draws some conclusions for the future relationship between the parties involved and suggests some changes that may improve the present situation. Publication Types: Review PMID: 10885119 [PubMed - indexed for MEDLINE] 2106: Br Med Bull. 2000;56(1):62-73. Genetically modified crops: methodology, benefits, regulation and public concerns. Halford NG, Shewry PR. IACR-Long Ashton Research Station, Department of Agricultural Sciences, University of Bristol, UK. The genetic modification of crop plants from the methodology involved in their production through to the current debate on their use in agriculture are reviewed. Techniques for plant transformation by Agrobacterium tumefaciens and particle bombardment, and for the selection of transgenic plants using marker genes are described. The benefits of currently available genetically modified (GM) crops in reducing waste and agrochemical use in agriculture, and the potential of the technology for further crop improvement in the future are discussed. The legal requirements for containment of novel GM crops and the roles of relevant regulatory bodies in ensuring that GM crops and food are safe are summarized. Some of the major concerns of the general public regarding GM crops and food: segregation of GM and non-GM crops and cross-pollination between GM crops and wild species, the use of antibiotic resistance marker genes, the prevention of new allergens being introduced in to the food chain and the relative safety of GM and non-GM foods are considered. Finally, the current debate on the use of GM crops in agriculture and the need for the government, scientists and industry to persevere with the technology in the face of widespread hostility is studied. Publication Types: Review PMID: 10885105 [PubMed - indexed for MEDLINE] 2107: Genetics. 2000 Jul;155(3):1161-74. Abnormal turning behavior in Drosophila larvae. Identification and molecular analysis of scribbler (sbb). Yang P, Shaver SA, Hilliker AJ, Sokolowski MB. Department of Zoology, University of Toronto, Mississauga, Ontario L5L 1C6, Canada. Our genetic dissection of behavior has isolated scribbler (sbb), a vital gene that encodes a novel protein expressed in the embryonic and larval nervous systems and in the imaginal discs. Larvae with mutations in sbb exhibit abnormally high amounts of turning behavior in the absence of food. sbb is a large gene spanning >50 kb of genomic DNA with four major developmentally regulated transcripts. Transgenic rescue of scribbler behavior was demonstrated by targeting expression of a normal sbb transgene (sbb(+)) expressing one of the major transcripts to the nervous system. The vital function of sbb was restored by ubiquitous expression of this transgene throughout development. PMID: 10880478 [PubMed - indexed for MEDLINE] 2108: Nature. 2000 Jun 22;405(6789):881. Comment on: Nature. 2000 Apr 20;404(6780):799. Biotech pioneers have duties as well as rights. Zhang BH. Publication Types: Comment Letter PMID: 10879507 [PubMed - indexed for MEDLINE] 2109: World Health Organ Tech Rep Ser. 2000;891:i-viii, 1-168. Evaluation of certain food additives. Fifty-first report of the Joint FAO/WHO Expert Committee on Food Additives. [No authors listed] This report presents the conclusions of a Joint FAO/WHO Expert Committee convened to evaluate the safety of various food additives, with a view to recommending Acceptable Daily Intakes (ADIs) for humans, and to prepare specifications for the identity and purity of food additives. The first part of the report contains a general discussion of the principles governing the toxicological evaluation of food additives, the assessment of intake of food additives, and the establishment and revision of specifications, including comments concerning enzyme preparations derived from genetically modified microorganisms and limits for heavy metals. A summary follows of the Committee's evaluations of toxicological data on specific food additives, including enzyme preparations (alpha-acetolactate decarboxylase and maltogenic amylase), flavouring agents (trans-anethole, furfural and menthol), food colours (curcumin and riboflavin from genetically modified Bacillus subtilis), glazing agents (medium- and low-viscosity mineral oils), preservatives (sulfur dioxide and sulfites), a sweetening agent (stevioside), thickening agents (carrageenan, processed Eucheuma seaweed and enzymatically hydrolysed sodium carboxymethyl cellulose), gamma-cyclodextrin, glucono-delta-lactone and the calcium, magnesium, potassium and sodium salts of gluconic acid, and polyglycitol syrup. The Committee also evaluated the safety of various groups of flavouring agents and assessed the intake of specific food additives, including benzoates, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), sulfites and tert-butylhydroquinone (TBHQ). Annexed to the report are tables summarizing the Committee's recommendations for ADIs of the food additives considered, changes in the status of specifications for these substances and specific flavouring agents, and further toxicological studies and other information required or desired. Publication Types: Technical Report PMID: 10876377 [PubMed - indexed for MEDLINE] 2110: Science. 2000 Jun 9;288(5472):1748-9. Comment on: Science. 1998 Aug 21;281(5380):1124-5. Science. 1999 Feb 19;283(5405):1094-5. Science. 1999 Oct 22;286(5440):656. Health risks of GM foods: many opinions but few data. Domingo JL. Publication Types: Comment Letter PMID: 10877692 [PubMed - indexed for MEDLINE] 2111: Yeast. 2000 Jun 15;16(8):675-729. Tailoring wine yeast for the new millennium: novel approaches to the ancient art of winemaking. Pretorius IS. Institute for Wine Biotechnology, University of Stellenbosch, Stellenbosch, ZA-7600, South Africa. isp@maties.sun.ac.za Yeasts are predominant in the ancient and complex process of winemaking. In spontaneous fermentations, there is a progressive growth pattern of indigenous yeasts, with the final stages invariably being dominated by the alcohol-tolerant strains of Saccharomyces cerevisiae. This species is universally known as the 'wine yeast' and is widely preferred for initiating wine fermentations. The primary role of wine yeast is to catalyze the rapid, complete and efficient conversion of grape sugars to ethanol, carbon dioxide and other minor, but important, metabolites without the development of off-flavours. However, due to the demanding nature of modern winemaking practices and sophisticated wine markets, there is an ever-growing quest for specialized wine yeast strains possessing a wide range of optimized, improved or novel oenological properties. This review highlights the wealth of untapped indigenous yeasts with oenological potential, the complexity of wine yeasts' genetic features and the genetic techniques often used in strain development. The current status of genetically improved wine yeasts and potential targets for further strain development are outlined. In light of the limited knowledge of industrial wine yeasts' complex genomes and the daunting challenges to comply with strict statutory regulations and consumer demands regarding the future use of genetically modified strains, this review cautions against unrealistic expectations over the short term. However, the staggering potential advantages of improved wine yeasts to both the winemaker and consumer in the third millennium are pointed out. Copyright 2000 John Wiley & Sons, Ltd. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 10861899 [PubMed - indexed for MEDLINE] 2112: Curr Opin Microbiol. 2000 Jun;3(3):276-82. Lactic acid bacteria: the bugs of the new millennium. Konings WN, Kok J, Kuipers OP, Poolman B. Department of Molecular Microbiology, Groningen Biomolecular Sciences and Biotechnology Institute, Biology Centre, University of Groningen, Haren, 9751 NN, The Netherlands. W.N.Konings@Biol.Rug.nl Lactic acid bacteria (LABs) are widely used in the manufacturing of fermented food and are among the best-studied microorganisms. Detailed knowledge of a number of physiological traits has opened new potential applications for these organisms in the food industry, while other traits might be beneficial for human health. Important new developments have been made in the research of LABs in the areas of multidrug resistance, bacteriocins and quorum sensing, osmoregulation, proteolysis, autolysins and bacteriophages. Recently, progress has been made in the construction of food-grade genetically modified LABs. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 10851157 [PubMed - indexed for MEDLINE] 2113: Anim Reprod Sci. 2000 Jul 2;60-61:161-72. Reproductive biotechnologies: current status in porcine reproduction. Day BN. Animal Science Unit, University of Missouri, College of Agriculture, Columbia, MO 65211, USA. dayb@missouri.edu During the past decade, considerable attention has been directed toward the development of reproductive technologies for both research purposes and for more controlled swine reproduction. Artificial insemination is an example of a technology that has continued to be expanded from early use in European countries to the USA and Canada where it is now estimated that a majority of the sows bred are artificially inseminated. In addition, several significant technological advancements have been made in the genetic modification of swine and interest has been generated in the possible use of swine as donors of specific tissues and of organs for the improvement of human health. At the same time, the systems for production of swine for human food continue to undergo major changes including, in some countries, the consolidation of swine into large, integrated units. These swine operations are very receptive to the use of technologies to reduce labor costs as well as a basis for increased production efficiency. Therefore, the combined interest in swine reproductive technologies by both the medical field and the swine industry creates an increased effort for the development of new technologies as well as for the implementation of existing ones. One of the more rapid technological advancements this decade has been the progress in in vitro production (IVP) of swine embryos. Major advancements have been made on the development of procedures for production of large numbers of embryos from oocytes collected at slaughter houses which are then matured (IVM) and fertilized (IVF) in the laboratory. Success in IVP has stimulated increased research in other areas that can be enhanced by the availability of embryos without a requirement for surgical collection from gilts or sows. One example is the combined use of IVF, gender-sorted sperm cells, and embryo transfer to produce offspring of a predicted sex. In a related area, instrumentation for non-surgical embryo transfer has recently been developed that results in significant improvement in this technology. Similar achievements have been gained in cryopreservation of embryos by vitrification. These developments will be reviewed with emphasis on the in vitro production of embryos from immature oocytes. Publication Types: Review PMID: 10844192 [PubMed - indexed for MEDLINE] 2114: Cancer Biother Radiopharm. 1999 Feb;14(1):1-4. Herceptin: breaking new ground. Cohen RL. Publication Types: Editorial PMID: 10850280 [PubMed - indexed for MEDLINE] 2115: Nat Biotechnol. 2000 Jun;18(6):670-4. Transgenic avidin maize is resistant to storage insect pests. Kramer KJ, Morgan TD, Throne JE, Dowell FE, Bailey M, Howard JA. Grain Marketing Production Research Center, Agricultural Research Service, US Department of Agriculture, Manhattan, KS 66502, USA. Avidin is a glycoprotein found in chicken egg white, that sequesters the vitamin biotin. Here we show that when present in maize at levels of > or =100 p.p.m., avidin is toxic to and prevents development of insects that damage grains during storage. Insect toxicity is caused by a biotin deficiency, as shown by prevention of toxicity with biotin supplementation. The avidin maize is not, however, toxic to mice when administered as the sole component of their diet for 21 days. These dates suggest that avidin expression in food or feed grain crops can be used as a biopesticide against a spectrum of stored-produce insect pests. PMID: 10835608 [PubMed - indexed for MEDLINE] 2116: Biotechnol Prog. 2000 May-Jun;16(3):435-41. Process options in hepatitis B surface antigen extraction from transgenic potato. Dogan B, Mason HS, Richter L, Hunter JB, Shuler ML. Department of Food Science, Boyce Thompson Institute, Cornell University, Ithaca, New York 14853, USA. The process conditions for recombinant hepatitis B surface antigen (HBsAg) extraction from transgenic potato were examined. The effects of temperature, the reducing agent beta-mercaptoethanol (BME), and proteinase inhibitors on the level of antigenic activity of recovered HBsAg were determined. Sedimentation profiles were performed to characterize HBsAg assembly into virus-like particles. Increasing the temperature of the sample for about 1 min increased the measured HBsAg antigenic activity. The optimum temperature was around 50 degrees C. A 3-fold enhancement of the antigenic activity was obtained in extract from transgenic potato expressing HBsAg, when monoclonal antibodies were used to assay for HBsAg. When antigenic activity was determined by polyclonal antibodies, no enhancement in the antigenic activity was obtained. Temperature may affect the conformation of the a epitope to which the monoclonal antibodies bind or alter the fluidity of surface lipid regions. BME increased the antigenic activity of HBsAg up to 4-fold when monoclonal antibodies directed against the a determinant were used, but there was no increase with polyclonal antibodies. This observation suggests that BME affects the structure or presentation of the a epitope. In the presence of BME and leupeptin, a proteinase inhibitor, higher antigenic activity was obtained. Leupeptin might protect the antigen, which might become more susceptible to proteolytic degradation after reduction, as a result of stimulation of sulfhydryl proteases. Although both temperature and BME increased the antigenic activity of HBsAg individually, when combined their interaction was antagonistic, resulting in reduced antigenic activity. Different proteinase inhibitors, including leupeptin, aprotinin, E-64, pefabloc, and pepstatin, had no significant effect on HBsAg from potato extract in a 2 h period in the absence of BME. The sedimentation profile of potato-produced HBsAg was determined in 5-30% sucrose gradients. Yeast-derived recombinant HBsAg was used as a positive control. The HBsAg from transgenic potato showed sedimentation and density properties that are very similar to the yeast-produced antigen, indicating assembly into virus-like particles. BME treatment did not change the sedimentation profile. Publication Types: Research Support, U.S. Gov't, Non-P.H.S. PMID: 10835246 [PubMed - indexed for MEDLINE] 2117: Food Chem Toxicol. 2000 Jun;38(6):473-83. Substantial equivalence of antinutrients and inherent plant toxins in genetically modified novel foods. Novak WK, Haslberger AG. Institute for Nutritional Sciences, University of Vienna, Austria. For a safety evaluation of foodstuff derived from genetically modified crops, the concept of the substantial equivalence of modified organisms with their parental lines is used following an environmental safety evaluation. To assess the potential pleiotropic effect of genetic modifications on constituents of modified crops data from US and EC documents were investigated with regard to inherent plant toxins and antinutrients. Analysed were documents of rape (glucosinolates, phytate), maize (phytate), tomato (tomatine, solanine, chaconine, lectins, oxalate), potato (solanine, chaconine, protease-inhibitors, phenols) and soybean (protease-inhibitors, lectins, isoflavones, phytate). In several documents used for notifications no declarations even on essential inherent plant toxins and antinutrients could be found, for instance data on phytate in modified maize were provided only in one of four documents. Significant variations in the contents of these compounds in parental and modified plants especially due to environmental influences were observed: drought stress, for example, was made responsible for significantly increased glucosinolate levels of up to 72.6micromol/g meal in modified and parental rape plants in field trials compared to recommended standard concentrations of less than 30micromol/g. Taking into account these wide natural variations generally the concentrations of inherent plant toxins and antinutrients in modified products were in the range of the concentrations in parental organisms. The results presented indicate that the concept of the substantial equivalence is useful for the risk assessment of genetically modified organisms (GMOs) used for novel foods but possible environmental influences on constituents of modified crops need more attention. Consistent guidelines, specifying data of relevant compounds which have to be provided for notification documents of specific organisms have to be established. Because of the importance of inherent plant toxins and antinutrients on nutritional safety, also coherent databases of standard parental lines and clear criteria for mandatory declarations are necessary. PMID: 10828499 [PubMed - indexed for MEDLINE] 2118: Med Confl Surviv. 2000 Jan-Mar;16(1):104-7. Feeding the world in the new millennium. Waterlow J. Department of Public Health and Policy, London School of Hygiene and Tropical Medicine. Food production per head in the world as a whole has begun to level off in the last decade, while the world population continues to grow, risking malnutrition, perhaps even starvation, civil unrest and environmental damage. Very little more land suitable for agriculture is available, and the factors behind the 'green revolution' may not produce further increases. Genetically modified crops offer the possibility of increased yields, but also present major problems. In developing countries, where yields are well below what is theoretically possible, the best approach may be better management by small farmers through improvements in their traditional methods. Much more financial support for and research in agriculture is needed, together with more equitable distribution of existing production and an end to trade practices designed to make the rich richer. PMID: 10824526 [PubMed - indexed for MEDLINE] 2119: Nature. 2000 May 11;405(6783):108. Comment in: Nature. 2000 Jun 15;405(6788):733. US reforms rules for telling public about GM food. Macilwain C. Publication Types: News PMID: 10821240 [PubMed - indexed for MEDLINE] 2120: Proc Nutr Soc. 1999 Nov;58(4):807-12. Social determinants of food choice. Shepherd R. Department of Psychology, University of Surrey, Guildford, UK. R.Shepherd@surrey.ac.uk Food choice is influenced by a large number of factors, including social and cultural factors. One method for trying to understand the impact of these factors is through the study of attitudes. Research is described which utilizes social psychological attitude models of attitude-behaviour relationships, in particular the Theory of Planned Behaviour. This approach has shown good prediction of behaviour, but there are a number of possible extensions to this basic model which might improve its utility. One such extension is the inclusion of measures of moral concern, which have been found to be important both for the choice of genetically-modified foods and also for foods to be eaten by others. It has been found to be difficult to effect dietary change, and there are a number of insights from social psychology which might address this difficulty. One is the phenomenon of optimistic bias, where individuals believe themselves to be at less risk from various hazards than the average person. This effect has been demonstrated for nutritional risks, and this might lead individuals to take less note of health education messages. Another concern is that individuals do not always have clear-cut attitudes, but rather can be ambivalent about food and about healthy eating. It is important, therefore, to have measures for this ambivalence, and an understanding of how it might impact on behaviour. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 10817147 [PubMed - indexed for MEDLINE] 2121: Plant Mol Biol. 2000 Mar;42(4):623-33. MDH1: an apple homeobox gene belonging to the BEL1 family. Dong YH, Yao JL, Atkinson RG, Putterill JJ, Morris BA, Gardner RC. Horticulture and Food Research Institute of New Zealand, Auckland. Differential display was used to isolate genes differentially expressed early in fruit development of apple (Malus domestica Borkh.). This approach resulted in the isolation of MDH1, a homeobox gene with a homeodomain similar to that of BELL1 (BEL1), which is involved in regulation of ovule development in Arabidopsis. However, outside the homeodomain MDH1 is quite different from BEL1. In apple, MDH1 mRNA was predominantly found in flowers, expanding leaves and expanding fruit. In pre-anthesis flowers, in situ hybridization showed that MDH1 mRNA accumulated in ovules. To further investigate the function of this new homeobox gene, MDH1 was transformed into Arabidopsis thaliana under the control of the cauliflower mosaic virus 35S promoter. The transgenic Arabidopsis plants showed dwarfing, reduced fertility and changes in carpel and fruit (silique) shape. The size and shape of the cells in the transgenic fruit was irregular. Both the transgenic phenotypes in Arabidopsis and the expression pattern of this gene in apple are consistent with the idea that MDH1 is likely to play an important role in control of plant fertility. Publication Types: Research Support, Non-U.S. Gov't PMID: 10809008 [PubMed - indexed for MEDLINE] 2122: Nat Biotechnol. 2000 May;18(5):551-4. Production of very-high-amylose potato starch by inhibition of SBE A and B. Schwall GP, Safford R, Westcott RJ, Jeffcoat R, Tayal A, Shi YC, Gidley MJ, Jobling SA. Unilever Research Colworth, Colworth House, Sharnbrook, Bedford MK44 1LQ, UK. High-amylose starch is in great demand by the starch industry for its unique functional properties. However, very few high-amylose crop varieties are commercially available. In this paper we describe the generation of very-high-amylose potato starch by genetic modification. We achieved this by simultaneously inhibiting two isoforms of starch branching enzyme to below 1% of the wild-type activities. Starch granule morphology and composition were noticeably altered. Normal, high-molecular-weight amylopectin was absent, whereas the amylose content was increased to levels comparable to the highest commercially available maize starches. In addition, the phosphorus content of the starch was increased more than fivefold. This unique starch, with its high amylose, low amylopectin, and high phosphorus levels, offers novel properties for food and industrial applications. PMID: 10802625 [PubMed - indexed for MEDLINE] 2123: Trends Biotechnol. 2000 Jun;18(6):232-3. Biotechnology food standards hunger for scientific rationale. Miller HI. Hoover Institution, Stanford University, Stanford, CA 94305-6010, USA. miller@hoover.stanford.edu PMID: 10802557 [PubMed - indexed for MEDLINE] 2124: Plant J. 2000 Apr;22(1):19-27. Functional analysis of HD2 histone deacetylase homologues in Arabidopsis thaliana. Wu K, Tian L, Malik K, Brown D, Miki B. Eastern Cereal and Oilseed Research Centre, Agriculture and Agri-Food Canada, Ottawa, Ontario, Canada K1A 0C6. Post-translational modification of histones, in particular acetylation, is an important mechanism in the regulation of eukaryotic gene expression. Histone deacetylases are enzymes that remove acetyl groups from the core histones and play a key role in the repression of transcription. HD2 is a maize histone deacetylase, which shows no sequence homology to the histone deacetylases identified from other eukaryotes. We have identified two putative HD2-like histone deacetylase cDNA clones, AtHD2A and AtHD2B, from Arabidopsis thaliana by screening the expressed sequence tag database. AtHD2A and AtHD2B encode putative proteins of 246 and 305 amino acids, and share 44% and 46% amino acid identity to the maize HD2, respectively. Northern blot analysis indicated that AtHD2A was highly expressed in flowers and young siliques of Arabidopsis plants, whereas AtHD2B was widely expressed in stems, leaves, flowers and young siliques. AtHD2A repressed transcription when directed to a promoter containing GAL4-binding sites as a GAL4 fusion protein. Deletion of the extended acidic domain or the domain containing predicted catalytic residues of AtHD2A resulted in the loss of gene repression activity, revealing the importance of both domains to AtHD2A function. Arabidopsis plants were transformed with a gene construct comprising an AtHD2A cDNA in the antisense orientation driven by a strong constitutive promoter, -394tCUP. Silencing of AtHD2A expression resulted in aborted seed development in transgenic Arabidopsis plants, suggesting that the AtHD2A gene product was important in the reproductive development of Arabidopsis thaliana. Publication Types: Research Support, Non-U.S. Gov't PMID: 10792817 [PubMed - indexed for MEDLINE] 2125: Sci Am. 2000 Apr;282(4):42-3. Rules of the game. Beardsley T. PMID: 10789246 [PubMed - indexed for MEDLINE] 2126: Poult Sci. 2000 Apr;79(4):525-7. Protein and energy evaluation of soybean meals processed from genetically modified high-protein soybeans. Edwards HM 3rd, Douglas MW, Parsons CM, Baker DH. Department of Animal Sciences, University of Illinois at Urbana-Champaign, 61801, USA. A conventional and two genetically modified soybean samples were processed to dehulled soybean meal (SBM) at a pilot plant and were compared with SBM from a commercial processing plant. Crude protein levels (%) of the experimental SBM samples were M700, 52.5; M702, 53.4; and M703, 62.7. The commercial SBM sample (UI) contained 47.5% protein. Amino acid, gross energy, lipid, and fiber analyses were carried out, and true metabolizable energy and true amino acid digestibility were determined with adult cecectomized cockerels. Digestible Lys, Met, Cys, Thr, and Val, and also TMEn, were higher (P < 0.05) and NDF, fat, and phospholipids were lower in M703 than in the other SBM samples. The results of this study indicate that M703 has considerable advantages over conventional SBM as a feed ingredient for broiler chickens. Publication Types: Research Support, Non-U.S. Gov't PMID: 10780648 [PubMed - indexed for MEDLINE] 2127: West J Med. 2000 Apr;172(4):220-1. The genetically modified food fight. Jacobson M. Center for Science in the Public Interest, Washington, DC 20009, USA. mjacobson@cspinet.org PMID: 10778361 [PubMed - indexed for MEDLINE] 2128: Science. 2000 Apr 14;288(5464):245-7. Comment in: Science. 2000 May 26;288(5470):1343. Biotechnology. Transgenic crops report fuels debate. Kaiser J. Publication Types: News PMID: 10777400 [PubMed - indexed for MEDLINE] 2129: Med J Aust. 2000 Feb 21;172(4):173-4. Comment in: Med J Aust. 2000 Feb 21;172(4):148-9. Genetically modified foods--food for thought. Leeder SR. Faculty of Medicine, University of Sydney, NSW. steve@medicine.usyd.edu.au We would be wise to hold off until we know more about the health, ecological and economic effects of genetically modified food. Publication Types: Review PMID: 10772590 [PubMed - indexed for MEDLINE] 2130: Med J Aust. 2000 Feb 21;172(4):170-3. Comment in: Med J Aust. 2000 Aug 7;173(3):166-7. Med J Aust. 2000 Aug 7;173(3):167. Med J Aust. 2000 Feb 21;172(4):148-9. Genetically modified foods--safety and regulatory issues. Huppatz JL, Fitzgerald PA. CSIRO Plant Industry, Canberra, ACT. Gene technology is a new form of biotechnology with much greater potential applications. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 10772589 [PubMed - indexed for MEDLINE] 2131: Med J Aust. 2000 Feb 21;172(4):148-9. Comment on: Med J Aust. 2000 Feb 21;172(4):170-3. Med J Aust. 2000 Feb 21;172(4):173-4. Genetically modified food: consternation, confusion, and crack-up. Horton R. Publication Types: Comment Editorial Review PMID: 10772580 [PubMed - indexed for MEDLINE] 2132: Trends Biotechnol. 2000 May;18(5):188-90. Genetically modified foods: economic aspects and public acceptance in Brazil. Oda LM, Soares BE. FIOCRUZ Biosafety Office, Department of Science and Biotechnology, Oswaldo Cruz Foundation, Rio de Janeiro, Brazil. oda@fiocruz.br Publication Types: Research Support, Non-U.S. Gov't Review PMID: 10758511 [PubMed - indexed for MEDLINE] 2133: J Biotechnol. 2000 Mar 31;78(3):251-8. Genomics, molecular genetics and the food industry. Pridmore RD, Crouzillat D, Walker C, Foley S, Zink R, Zwahlen MC, Brüssow H, Pétiard V, Mollet B. Nestec Ltd., Nestlé Research Center, Vers-chez-les-Blanc, 1000, Lausanne, Switzerland. raymond-david.primore@rdls.nestle.com The production of foods for an increasingly informed and selective consumer requires the coordinated activities of the various branches of the food chain in order to provide convenient, wholesome, tasty, safe and affordable foods. Also, the size and complexity of the food sector ensures that no single player can control a single process from seed production, through farming and processing to a final product marketed in a retail outlet. Furthermore, the scientific advances in genome research and their exploitation via biotechnology is leading to a technology driven revolution that will have advantages for the consumer and food industry alike. The segment of food processing aids, namely industrial enzymes which have been enhanced by the use of biotechnology, has proven invaluable in the production of enzymes with greater purity and flexibility while ensuring a sustainable and cheap supply. Such enzymes produced in safe GRAS microorganisms are available today and are being used in the production of foods. A second rapidly evolving segment that is already having an impact on our foods may be found in the new genetically modified crops. While the most notorious examples today were developed by the seed companies for the agro-industry directed at the farming sector for cost saving production of the main agronomical products like soya and maize, its benefits are also being seen in the reduced use of herbicides and pesticides which will have long term benefits for the environment. Technology-driven advances for the food processing industry and the consumer are being developed and may be divided into two separate sectors that will be presented in greater detail: 1. The application of genome research and biotechnology to the breeding and development of improved plants. This may be as an aid for the cataloging of industrially important plant varieties, the rapid identification of key quality traits for enhanced classical breeding programs, or the genetic modification of important plants for improved processing properties or health characteristics. 2. The development of advanced microorganisms for food fermentations with improved flavor production, health or technological characteristics. Both yeasts and bacteria have been developed that fulfill these requirements, but are as yet not used in the production of foods. PMID: 10751686 [PubMed - indexed for MEDLINE] 2134: CMAJ. 2000 Mar 21;162(6):874. Precautionary principle leads to "may contain" clause for genetically modified foods. Pinker S. Publication Types: News PMID: 10750480 [PubMed - indexed for MEDLINE] 2135: Nat Biotechnol. 2000 Apr;18(4):375. GM food singled out for labeling in the US. Fox JL. Publication Types: News PMID: 10748509 [PubMed - indexed for MEDLINE] 2136: Plant Mol Biol. 1999 Dec;41(6):777-83. Expression of fungal thermotolerant endo-1,4-beta-glucanase in transgenic barley seeds during germination. Nuutila AM, Ritala A, Skadsen RW, Mannonen L, Kauppinen V. VTT Biotechnology and Food Research, Espoo, Finland. The malting quality of two barley cultivars, Kymppi and Golden Promise, was modified to better meet the requirements of the brewing process. The egl1 gene, coding for fungal thermotolerant endo-1,4-beta-glucanase (EGI, cellulase), was transferred to the cultivars using particle bombardment, and transgenic plants were regenerated on bialaphos selection. Integration of the egl1 gene was confirmed by Southern blot hybridization. The transgenic seeds were screened for the expression of the heterologous EGI. Under the high-pI alpha-amylase promoter, the egl1 gene was expressed during germination. The heterologous enzyme was thermotolerant at 65 degrees C for 2 h, thus being suitable for mashing conditions. The amount of heterologous EGI produced by the seeds (ca. 0.025% of soluble seed protein), has been shown to be sufficient to reduce wort viscosity by decreasing the soluble beta-glucan content. A decrease in the soluble beta-glucan content in the wort improves the filtration rate of beer. PMID: 10737142 [PubMed - indexed for MEDLINE] 2137: Adv Exp Med Biol. 1999;467:749-55. Approaches in novel feed--EU legislation. Petersen U. Bundesministerium für Ernährung, Landwirtschaft und Forsten, Bonn, Germany. The relevant EC-legislation in the field of novel feed, inclusive the legislation about genetically modified micro-organisms is presented and discussed. PMID: 10721128 [PubMed - indexed for MEDLINE] 2138: Science. 2000 Feb 25;287(5457):1390. Ecology. New corn plant draws fire from GM food opponents. Ferber D. Publication Types: News PMID: 10722381 [PubMed - indexed for MEDLINE] 2139: J Biotechnol. 2000 Jan 28;77(1):103-14. Chemical fingerprinting for the evaluation of unintended secondary metabolic changes in transgenic food crops. Noteborn HP, Lommen A, van der Jagt RC, Weseman JM. DLO-State Institute for Quality Control of Agricultural Products (RIKILT-DLO), Wageningen, The Netherlands. h.p.j.m.noteborn@rikilt.dlo.nl A common element in designed guidelines for assessment of the food safety of transgenic crops is centred on a comparative analytical analysis with conventionally bred crop plants, assuming that these products have a long history of safe use (i.e. OECD-principle of substantial equivalence). In this study we examine the utility of an off-line combination of 400 MHz proton (1H)-NMR spectroscopy and liquid chromatography (LC) for the multi-component comparison of low-molecular weight compounds (i.e. chemical fingerprinting) in complex plant matrices. The developed NMR-methodology can contribute to the demonstration of substantial equivalence by its ability to compare possible compositional alterations in a novel food crop with respect to related non-transgenic reference lines. In this respect a hierarchical approach is proposed by comparing the chemical fingerprints of the transgenic crop plant to those of: (1) isogenic parental or closely related lines bred at identical and multiple sites; (2) extended ranges of commercial varieties of that plant; and (3) downstream processing effects. This is of importance to assess the likelihood that some of the statistical differences in a transgenic crop plant may be false positives due to chance alone or arose from natural genetic and/or physiologic variations. Publication Types: Research Support, Non-U.S. Gov't PMID: 10674217 [PubMed - indexed for MEDLINE] 2140: Dermatol Online J. 1999 Nov;5(2):1. Insulin-like growth factor 1 and hair growth. Su HY, Hickford JG, Bickerstaffe R, Palmer BR. Division of Molecular Neurobiology, University of Cincinnati College of Medicine, P.O. Box 670559, Cincinnati, OH 45267, USA. Insulin-like growth factor 1 (IGF-1) has been identified as an important growth factor in many biological systems.[1] It shares considerable structural homology with insulin and exerts insulin-like effects on food intake and glucose metabolism. Recently it has been suggested to play a role in regulating cellular proliferation and migration during the development of hair follicles. [2,3] To exert its biological effects, the IGF-1 is required to activate cells by binding to specific cell-surface receptors. The type I IGF receptor (IGF-1R) is the only IGF receptor to have IGF-mediated signaling functions.[1] In circulation, this growth factor mediates endocrine action of growth hormone (GH) on somatic growth and is bound to specific binding proteins (BPs). The latter control IGF transport, efflux from vascular compartments and association with cell surface receptors.[4] In tissues, IGF-1 is produced by mesenchymal type cells and acts in a paracrine and autocrine fashion by binding to the IGF-1R. This binding activates the receptor tyrosine kinase (RTK) that triggers the downstream responses and finally stimulates cell division.[5] IGF-1 may therefore be able to stimulate the proliferation of hair follicle cells through cellular signaling pathways of its receptors. Local infusion of IGF-1 into sheep has been reported to be capable of stimulating protein synthesis in the skin.[6] It may also increase the production of wool keratin. Recently, transgenic mice overexpressing IGF-1 in the skin have been shown to have earlier hair follicle development than controls.[7] In addition, this growth factor plays an important role in many cell types as a survival factor to prevent cell death.[8] This anti-apoptotic function of IGF-1 may be important to the development of follicle cells as follicles undergo a growth cycle where the regressive, catagen phase is apoptosis driven. In this review, the effects of IGF-1 on follicle cell proliferation and differentiation are discussed. In particular, the paracrine versus endocrine action of IGF-1 on hair growth and the targeting of expression of the growth factor to the follicles of transgenic animals will be emphasized. The anti-apoptotic role of IGF-1 in hair follicles is also reviewed. Prospects for future studies on hair and fiber growth by IGF-1 are discussed. PMID: 10673454 [PubMed - indexed for MEDLINE] 2141: Science. 2000 Jan 21;287(5452):431-2. Policy forum: genetic technologies. Monitoring and labeling for genetically modified products. Haslberger AG. Federal Chancellory, Vienna, Austria. hasi1@via.at PMID: 10671171 [PubMed - indexed for MEDLINE] 2142: An Esp Pediatr. 1999 Dec;51(6):617-21. [Transgenic food] [Article in Spanish] Ballabriga A, Moya M. PMID: 10666893 [PubMed - indexed for MEDLINE] 2143: Lancet. 2000 Jan 29;355(9201):414. Comment in: Lancet. 2000 Mar 11;355(9207):931. Do genetically modified foods affect human health? Godfrey J. Publication Types: Letter PMID: 10665592 [PubMed - indexed for MEDLINE] 2144: Time. 1999 Nov 29;154(22):49-50. Who's afraid of Frankenfood? Golden F. Publication Types: News PMID: 10662098 [PubMed - indexed for MEDLINE] 2145: Nat Biotechnol. 2000 Feb;18(2):143. Erratum in: Nat Biotechnol 2000 May;18(5):473. FDA, researchers consider first transgenic fish. Niiler E. Publication Types: News Research Support, U.S. Gov't, Non-P.H.S. PMID: 10657111 [PubMed - indexed for MEDLINE] 2146: Carcinogenesis. 2000 Feb;21(2):265-73. DNA adduct formation and molecular analysis of in vivo lacI mutations in the mammary tissue of Big Blue rats treated with 7, 12-dimethylbenz[a]anthracene. Manjanatha MG, Shelton SD, Culp SJ, Blankenship LR, Casciano DA. Department of Health and Human Services, Food and Drug Administration, National Center for Toxicological Research, Divisions of Genetic and Biochemical Toxicology, Jefferson, AR 72079, USA. mmanjanatha@nctr.fda.gov Recently we compared the lacI and Hprt mutant frequencies (MFs) and types of mutations in lymphocytes of Big Blue((R)) (BB) rats exposed to 7,12-dimethylbenz[a]anthracene (DMBA) under conditions that result in mammary gland tumors. In this study, we have examined the target mammary tissue for DMBA-induced DNA adducts, lacI MF and types of lacI mutations. Seven-week-old female BB rats were given single doses of 0, 20 or 130 mg/kg DMBA by gavage and the DNA adducts and lacI MFs in the mammary tissue were measured over a period of 14 days and 18 weeks, respectively, following treatment. The lacI MF in the mammary tissue increased for 10 weeks and then remained relatively constant; 130 mg/kg DMBA produced a 14-fold increase in the MF (255 +/- 50 x 10(-6) p.f.u.) over control MF (18. 3 +/- 4 x 10(-6) p.f.u.). (32)P-post-labeling analysis of DNA from mammary tissue and splenic lymphocytes of treated rats revealed two major adducts. Comparison of these adducts with DMBA standards indicated that the adducts formed by DMBA involved both G:C and A:T base pairs. DNA sequencing revealed that the majority of DMBA-induced lacI mutations were base pair substitutions and that A:T-->T:A (44% of the independent mutations) and G:C-->T:A (24% of the independent mutations) transversions were the predominant types. Furthermore, the mutational results revealed a 'hotspot' for a G-->T mutation in codon 95 (GTG-->TTG) of the lacI gene in mammary tissue. These results suggest that DMBA is highly mutagenic to lacI in mammary tissue and that adducts with both G:C and A:T base pairs participate in forming mutations in DMBA-treated BB rats. Publication Types: Comparative Study PMID: 10657967 [PubMed - indexed for MEDLINE] 2147: Vopr Pitan. 1999;68(5-6):3-8. [Current approaches to the evaluation of genetically modified food products. Soybean 40-3-2 data] [Article in Russian] Onishchenko GG, Tutel'ian VA, Petukhov AI, Korolev AA, Aksiuk IN, Sorokina EIu. Different methodological approaches were elaborated to evaluate quality and safety of genetically modified food products. The new engineering is proposed to rate medical, biological, genetic and technological advantage of these products. Using the same engineering, a complete analysis of the genetically modified soybean 40-3-2 ("Monsanto Co", USA) has been performed. Publication Types: Comparative Study English Abstract PMID: 10641272 [PubMed - indexed for MEDLINE] 2148: Biosci Biotechnol Biochem. 1999 Nov;63(11):1942-6. Safety assessment of transgenic potatoes with soybean glycinin by feeding studies in rats. Hashimoto W, Momma K, Yoon HJ, Ozawa S, Ohkawa Y, Ishige T, Kito M, Utsumi S, Murata K. Research Institute for Food Science, Kyoto University, Japan. hasimoto@food2.food.kyoto-u.ac.jp Feeding studies of transgenic potatoes with native and designed soybean glycinins in rats were done for four weeks. The designed glycinin has four additional methioninyl residues in the middle of the glycinin molecule. Rats were divided into four groups fed (I) only a commercial diet, (II) the diet plus non-transgenic potatoes, (III) the diet plus transgenic potatoes with native glycinin, and (IV) the diet plus transgenic potatoes with designed glycinin. Rats were fed 2,000 mg/kg-weight potatoes every day by oral administration. During the period tested, rats in each group (groups II, III, and IV) grew well without marked differences in appearance, food intake, body weight, or in cumulative body weight gain. No significant differences were also found in blood count, blood composition, and in internal organ weights among the rats after feeding potatoes (groups II, III, and IV) for four weeks. Necropsy at the end of experiment indicated neither pathologic symptoms in all rats tested nor histopathological abnormalities in liver and kidney. Judging from these results, the transgenic potatoes with glycinins are confirmed to have nearly the same nutritional and biochemical characteristics as non-transgenic one. Publication Types: Comparative Study Research Support, Non-U.S. Gov't PMID: 10635558 [PubMed - indexed for MEDLINE] 2149: Science. 2000 Jan 14;287(5451):303-5. Comment in: Science. 2000 Jan 14;287(5451):241-3. Science. 2000 Jun 16;288(5473):1966-7. Engineering the provitamin A (beta-carotene) biosynthetic pathway into (carotenoid-free) rice endosperm. Ye X, Al-Babili S, Klöti A, Zhang J, Lucca P, Beyer P, Potrykus I. Institute for Plant Sciences, Swiss Federal Institute of Technology, CH-8092 Zurich, Switzerland. University of Freiburg, Center for Applied Biosciences, D-79104 Freiburg, Germany. Rice (Oryza sativa), a major staple food, is usually milled to remove the oil-rich aleurone layer that turns rancid upon storage, especially in tropical areas. The remaining edible part of rice grains, the endosperm, lacks several essential nutrients, such as provitamin A. Thus, predominant rice consumption promotes vitamin A deficiency, a serious public health problem in at least 26 countries, including highly populated areas of Asia, Africa, and Latin America. Recombinant DNA technology was used to improve its nutritional value in this respect. A combination of transgenes enabled biosynthesis of provitamin A in the endosperm. Publication Types: Research Support, Non-U.S. Gov't PMID: 10634784 [PubMed - indexed for MEDLINE] 2150: JAMA. 2000 Jan 12;283(2):188-90. Genetically modified crops feed ongoing controversy. Marwick C. Publication Types: News PMID: 10634325 [PubMed - indexed for MEDLINE] 2151: Trends Biotechnol. 2000 Jan;18(1):8-10. Ethical issues in biotechnology. Polkinghorne JC. Queens' College, University of Cambridge, UK. New ethical questions have arisen from our ability to intervene in the structure of the genome. Responsible use of this technique requires ethical evaluation in which experts, potential beneficiaries and the general public should all participate. The examples of genetically modified food and of human genetics help to illustrate the issues involved. PMID: 10631772 [PubMed - indexed for MEDLINE] 2152: Appetite. 1999 Dec;33(3):343-60. Consumers' cognitions with regard to genetically modified foods. Results of a qualitative study in four countries. Bredahl L. MAPP, The Aarhus School of Business, Denmark. lone.bredahl@mar.hha.dk The objective of this research was to gain insight into consumers>> attitudes towards genetic modification in food production. With means-end chain theory as the theoretical basis, laddering interviews were conducted with 400 consumers in Denmark, Germany, the United Kingdom and Italy. Perceived risks and benefits of genetic modification in foods were investigated using beer and yoghurt as examples. German and Danish responses revealed more complex cognitive structures than did the results from the United Kingdom and Italy. In all four countries, however, applying genetic modification was associated with unnaturalness and low trustworthiness of the resulting products, independently of whether the genetically modified material was traceable in the product. Moral considerations were voiced as well, as were a number of other consequences that were perceived to conflict with both individual and social values. Copyright 1999 Academic Press. Publication Types: Multicenter Study Research Support, Non-U.S. Gov't PMID: 10625527 [PubMed - indexed for MEDLINE] 2153: Biochem Biophys Res Commun. 2000 Jan 7;267(1):466-72. Growth efficiency in transgenic tilapia (Oreochromis sp.) carrying a single copy of an homologous cDNA growth hormone. Martínez R, Juncal J, Zaldívar C, Arenal A, Guillén I, Morera V, Carrillo O, Estrada M, Morales A, Estrada MP. Division of Mammalian Cell Genetics, Centro de Ingeniería Genética y Biotecnología, Havana, Cuba. Growth hormone (GH) has been shown to have a profound impact on fish physiology and metabolism. However, detailed studies in transgenic fish have not been conducted. We have characterized the food conversion efficiency, protein profile, and biochemical correlates of growth rate in transgenic tilapia expressing the tilapia GH cDNA under the control of human cytomegalovirus regulatory sequences. Transgenic tilapia exhibited about 3.6-fold less food consumption than nontransgenic controls (P < 0.001). The food conversion efficiency was significantly (P < 0.05) higher (290%) in transgenic tilapia (2.3 +/- 0.4) than in the control group (0.8 +/- 0.2). Efficiency of growth, synthesis retention, anabolic stimulation, and average protein synthesis were higher in transgenic than in nontransgenic tilapia. Distinctive metabolic differences were found in transgenic juvenile tilapia. We had found differences in hepatic glucose, and in agreement with previous results we observed differences in the level of enzymatic activities in target organs. We conclude that GH-transgenic juvenile tilapia show altered physiological and metabolic conditions and are biologically more efficient. Copyright 2000 Academic Press. PMID: 10623643 [PubMed - indexed for MEDLINE] 2154: Genome Res. 1999 Dec;9(12):1159-62. Public concerns over transgenic crops. Dale PJ. John Innes Centre, Norwich NR4 7UH, UK. phil.dale@bbsrc.ac.uk Publication Types: Review PMID: 10613837 [PubMed - indexed for MEDLINE] 2155: Science. 1999 Nov 26;286(5445):1662-6. Comment in: Science. 2000 Feb 4;287(5454):803-4. Science. 2000 Jan 7;287(5450):41. GM crops in the cross hairs. Ferber D. Publication Types: News PMID: 10610557 [PubMed - indexed for MEDLINE] 2156: Science. 1999 Nov 26;286(5445):1666-8. Ag biotech moves to mollify its critics. Enserink M. Publication Types: News PMID: 10610558 [PubMed - indexed for MEDLINE] 2157: J Agric Food Chem. 1999 Dec;47(12):5261-6. Real-time quantitative PCR detection of genetically modified Maximizer maize and Roundup Ready soybean in some representative foods. Vaïtilingom M, Pijnenburg H, Gendre F, Brignon P. Tepral, Beverage Division Research Center of Danone Group, 68 Route d'Oberhausbergen, 67037 Strasbourg Cedex, France. A fast and quantitative method was developed to detect transgenic "Maximizer" maize "event 176" (Novartis) and "Roundup Ready" soybean (Monsanto) in food by real-time quantitative PCR. The use of the ABI Prism 7700 sequence detection system allowed the determination of the amplified product accumulation through a fluorogenic probe (TaqMan). Fluorescent dyes were chosen in such a way as to coamplify total and transgenic DNA in the same tube. Using real-time quantitative PCR, 2 pg of transgenic or total DNA per gram of starting sample was detected in 3 h after DNA extraction and the relative amounts of "Maximizer" maize and "Roundup Ready" soybean in some representative food products were quantified. PMID: 10606606 [PubMed - indexed for MEDLINE] 2158: J Agric Food Chem. 1999 Dec;47(12):5038-43. Erratum in: J Agric Food Chem 2001 Jul;49(7):3508. Genetically modified organisms in food-screening and specific detection by polymerase chain reaction. Vollenhofer S, Burg K, Schmidt J, Kroath H. Austrian Research Centers Seibersdorf, Biotechnology Unit, A-2444 Seibersdorf, Austria. sabine.vollenhofer@arcs.ac.at PCR methods for the detection of genetically modified organisms (GMOs) were developed that can be used for screening purposes and for specific detection of glyphosate-tolerant soybean and insect-resistant maize in food. Primers were designed to amplify parts of the 35S promoter derived from Cauliflower Mosaic Virus, the NOS terminator derived from Agrobacterium tumefaciens and the antibiotic marker gene NPTII (neomycin-phosphotransferase II), to allow for general screening of foods. PCR/hybridization protocols were established for the detection of glyphosate-tolerant RoundUp Ready soybean and insect-resistant Bt-maize. Besides hybridization, confirmation of the results using restriction analysis was also possible. The described methods enabled a highly sensitive and specific detection of GMOs and thus provide a useful tool for routine analysis of raw and processed food products. PMID: 10606569 [PubMed - indexed for MEDLINE] 2159: Nature. 1999 Dec 9;402(6762):571. US food-safety body hears protests over genetically modified food. Macilwain C. Publication Types: News PMID: 10604452 [PubMed - indexed for MEDLINE] 2160: Nature. 1999 Dec 9;402(6762):566. Indian patent law will protect crops 'unless injurious to health'. Jayaraman KS. Publication Types: News PMID: 10604445 [PubMed - indexed for MEDLINE] 2161: Mol Gen Genet. 1999 Oct;262(3):453-61. Isolation and functional characterisation of a novel type of carotenoid biosynthetic gene from Xanthophyllomyces dendrorhous. Verdoes JC, Krubasik KP, Sandmann G, van Ooyen AJ. Department of Food Technology and Nutritional Sciences, Wageningen University, The Netherlands. Jan.Verdoes@imb.ftns.wau.nl The red heterobasidiomycetous yeast Xanthophyllomyces dendrorhous (perfect state of Phaffia rhodozyma) contains a novel type of carotenoid biosynthetic enzyme. Its structural gene, designated crtYB, was isolated by functional complementation in a genetically modified, carotenogenic Escherichia coli strain. Expression studies in different carotenogenic E. coli strains demonstrated that the crt YB gene encodes a bifunctional protein involved both in synthesis of phytoene from geranylgeranyl diphosphate and in cyclisation of lycopene to beta-carotene. By sequence comparison with other phytoene synthases and complementation studies in E. coli with various deletion derivatives of the crtYB gene, the regions responsible for phytoene synthesis and lycopene cyclisation were localised within the protein. Publication Types: Comparative Study Research Support, Non-U.S. Gov't PMID: 10589832 [PubMed - indexed for MEDLINE] 2162: Nature. 1999 Nov 18;402(6759):231-2. Comment in: Nature. 2000 Mar 16;404(6775):222. Much food, many problems. Trewavas A. Institute of Cell and Molecular Biology, University of Edinburgh, UK. PMID: 10580487 [PubMed - indexed for MEDLINE] 2163: Nature. 1999 Nov 18;402(6759):229. Comment on: Nature. 1999 Oct 7;401(6753):525-6. Genetically modified foods face rigorous safety evaluation. Gasson MJ. Publication Types: Comment Letter PMID: 10580485 [PubMed - indexed for MEDLINE] 2164: Nature. 1999 Oct 28;401(6756):831-2. Developing countries look for guidance in GM crops debate... Macilwain C. Publication Types: Congresses News PMID: 10553887 [PubMed - indexed for MEDLINE] 2165: Nature. 1999 Oct 28;401(6756):829. Collaborations essential for food in the developing world. [No authors listed] Publication Types: Congresses Editorial PMID: 10553886 [PubMed - indexed for MEDLINE] 2166: Science. 1999 Oct 22;286(5440):656. Comment in: Science. 2000 Jun 9;288(5472):1748-9. Transgenic food debate. The Lancet scolded over Pusztai paper. Enserink M. Publication Types: News PMID: 10577214 [PubMed - indexed for MEDLINE] 2167: Lancet. 1999 Nov 13;354(9191):1729. Comment on: Lancet. 1999 May 29;353(9167):1811. Lancet. 1999 Oct 16;354(9187):1314-5. Lancet. 1999 Oct 16;354(9187):1353-4. GM food debate. Klug A. Publication Types: Comment Letter PMID: 10568596 [PubMed - indexed for MEDLINE] 2168: Lancet. 1999 Nov 13;354(9191):1729. Comment on: Lancet. 1999 Oct 16;354(9187):1314-5. GM food debate. Fisken RA. Publication Types: Comment Letter PMID: 10568595 [PubMed - indexed for MEDLINE] 2169: Lancet. 1999 Nov 13;354(9191):1729. Comment on: Lancet. 1999 Oct 16;354(9187):1314-5. Lancet. 1999 Oct 16;354(9187):1353-4. GM food debate. Feldbaum CB. Publication Types: Comment Letter PMID: 10568594 [PubMed - indexed for MEDLINE] 2170: Lancet. 1999 Nov 13;354(9191):1728; author reply 1728-9. Comment on: Lancet. 1999 Oct 16;354(9187):1353-4. Lancet. 1999 Oct 16;354(9187):1354-5. GM food debate. Kilpatrick DC. Publication Types: Comment Letter PMID: 10568593 [PubMed - indexed for MEDLINE] 2171: Lancet. 1999 Nov 13;354(9191):1727-8; author reply 1728-9. Comment on: Lancet. 1999 Oct 16;354(9187):1354-5. GM food debate. Munro S. Publication Types: Comment Letter PMID: 10568592 [PubMed - indexed for MEDLINE] 2172: Lancet. 1999 Nov 13;354(9191):1726; author reply 1726-7. Comment on: Lancet. 1999 Oct 16;354(9187):1353-4. GM food debate. Lachmann P. Publication Types: Comment Letter PMID: 10568591 [PubMed - indexed for MEDLINE] 2173: Plant Mol Biol. 1999 Sep;41(1):45-55. A tobacco cryptic constitutive promoter, tCUP, revealed by T-DNA tagging. Foster E, Hattori J, Labbé H, Ouellet T, Fobert PR, James LE, Iyer VN, Miki BL. Eastern Cereal and Oilseed Research Centre, Agriculture and Agri-Food Canada, Ottawa, Ontario. We have isolated a constitutive promoter sequence, tCUP, from tobacco by T-DNA tagging using a promoterless GUS-nos3' reporter gene construct. The T-DNA integration event produced a translational fusion with the GUS gene that is expressed widely in organs, at both the mRNA and enzyme activity levels. In tobacco transformed with a tCUP-GUS-nos3' gene, GUS specific activity in leaves was within a range of values similar to those of plants transformed with the widely used constitutive promoter gene fusion, CaMV 35S promoter-GUS-nos3'. Characteristics of the tCUP promoter sequence differ from those of other plant constitutive promoters; for instance, the tCUP sequence lacks a TATA box. Transcription initiates at a single site within the tCUP sequence which is similar to a transcriptional start site consensus sequence determined for plant genes. The tCUP promoter is cryptic as RNA accumulation at the transcriptional start site is not detected in untransformed tobacco. Thus, tCUP is the first example of a cryptic, constitutive promoter isolated from plants. The tCUP-GUS-nos3' gene fusion produced GUS activity in tissues of all species tested suggesting that tCUP may utilize fundamental transcription mechanisms found in plants. PMID: 10561067 [PubMed - indexed for MEDLINE] 2174: Development. 1999 Dec;126(23):5387-98. Control of DAF-7 TGF-(alpha) expression and neuronal process development by a receptor tyrosine kinase KIN-8 in Caenorhabditis elegans. Koga M, Take-uchi M, Tameishi T, Ohshima Y. Department of Biology, Graduate School of Sciences, Kyushu University, Fukuoka, Japan. KIN-8 in C. elegans is highly homologous to human ROR-1 and 2 receptor tyrosine kinases of unknown functions. These kinases belong to a new subfamily related to the Trk subfamily. A kin-8 promoter::gfp fusion gene was expressed in ASI and many other neurons as well as in pharyngeal and head muscles. A kin-8 deletion mutant was isolated and showed constitutive dauer larva formation (Daf-c) phenotype: about half of the F(1) progeny became dauer larvae when they were cultivated on an old lawn of E. coli as food. Among the cells expressing kin-8::gfp, only ASI sensory neurons are known to express DAF-7 TGF-(beta), a key molecule preventing dauer larva formation. In the kin-8 deletion mutant, expression of daf-7::gfp in ASI was greatly reduced, dye-filling in ASI was specifically lost and ASI sensory processes did not completely extend into the amphid pore. The Daf-c phenotype was suppressed by daf-7 cDNA expression or a daf-3 null mutation. ASI-directed expression of kin-8 cDNA under the daf-7 promoter or expression by a heat shock promoter rescued the dye-filling defect, but not the Daf-c phenotype, of the kin-8 mutant. These results show that the kin-8 mutation causes the Daf-c phenotype through reduction of the daf-7 gene expression and that KIN-8 function is cell-autonomous for the dye-filling in ASI. KIN-8 is required for the process development of ASI, and also involved in promotion of daf-7 expression through a physiological or developmental function. Publication Types: Research Support, Non-U.S. Gov't PMID: 10556063 [PubMed - indexed for MEDLINE] 2175: Nat Biotechnol. 1999 Nov;17(11):1137-8. Quantitation of genetically modified organisms in food. Hübner P, Studer E, Lüthy J. Official food control authority of the Canton of Zurich, Switzerland. klzh@bluewin.ch PMID: 10545927 [PubMed - indexed for MEDLINE] 2176: Nat Biotechnol. 1999 Nov;17(11):1047. GMO roundup. Hodgson J. Publication Types: News PMID: 10545872 [PubMed - indexed for MEDLINE] 2177: Nat Biotechnol. 1999 Nov;17(11):1042-3. Substantial equivalence: its uses and abuses. Miller HI. Hoover Institution, Stanford University, Stanford, CA 94305-6010, USA. miller@hoover.stanford.edu PMID: 10545866 [PubMed - indexed for MEDLINE] 2178: US News World Rep. 1999 Jul 26;127(4):38-41. The curse of Frankenfood. Genetically modified crops stir up controversy at home and abroad. Longman PJ. Publication Types: News PMID: 10539693 [PubMed - indexed for MEDLINE] 2179: Lancet. 1999 Oct 16;354(9187):1315-6. Comment in: Lancet. 1999 Oct 16;354(9187):1314-5. Comment on: Lancet. 1999 Oct 16;354(9187):1353-4. Adequacy of methods for testing the safety of genetically modified foods. Kuiper HA, Noteborn HP, Peijnenburg AA. RIKILT (National Institute for Quality Control of Agricultural Products), Wageningen University and Research Centre, The Netherlands. Publication Types: Comment PMID: 10533854 [PubMed - indexed for MEDLINE] 2180: Lancet. 1999 Oct 16;354(9187):1314-5. Comment in: Lancet. 1999 Nov 13;354(9191):1729. Lancet. 1999 Nov 13;354(9191):1729. Lancet. 1999 Nov 13;354(9191):1729. Lancet. 2001 Jan 27;357(9252):309-10. Comment on: Lancet. 1999 Oct 16;354(9187):1315-6. Lancet. 1999 Oct 16;354(9187):1353-4. Lancet. 1999 Oct 16;354(9187):1354-5. Genetically modified foods: "absurd" concern or welcome dialogue? Horton R. The Lancet, London, UK. Publication Types: Comment PMID: 10533853 [PubMed - indexed for MEDLINE] 2181: Harv Mens Health Watch. 1999 Oct;4(3):1-3. High-tech dining: is America ready? [No authors listed] PMID: 10529293 [PubMed - indexed for MEDLINE] 2182: Alcohol Alcohol. 1999 Sep-Oct;34(5):690-8. Effects of Hypericum perforatum extraction on alcohol intake in Marchigian Sardinian alcohol-preferring rats. Perfumi M, Ciccocioppo R, Angeletti S, Cucculelli M, Massi M. Department of Pharmacological Sciences and Experimental Medicine, University of Camerino, Italy. The present study investigated the effect of acute intragastric (i.g.) administration of dry Hypericum perforatum extract (HPE), containing 0.3% hypericin, on ethanol intake in genetically selected Marchigian Sardinian alcohol-preferring (msP) rats. The i.g. administration of HPE, 125 or 250 mg/kg, induced a 30-40% reduction in ethanol intake in rats offered 10% (v/v) ethanol for 2 h/day. The effect of these doses was selective, since they modified neither food intake nor food-associated drinking; neither did the same doses modify the rat's gross behaviour in the open-field test. A dose of 500 mg/kg frequently induced immobility and a general suppression of ingestive behaviour. In rats offered 10% ethanol for 12 h/day, ethanol intake following treatment with 250 mg/kg HPE was significantly lower than that of controls for up to 10 h. The effect on ethanol intake was not related to the antidepressant-like effect of HPE revealed in the forced swimming test. In this regard, the effect on ethanol intake was observed after a single administration of 125 mg/kg, whereas the antidepressant effect was observed only after repeated treatment with doses higher than 125 mg/kg HPE. The i.g. administration of HPE, 250 mg/kg, did not affect blood-alcohol levels following i.g. treatment with 0.7 g/kg ethanol, the amount usually ingested in a single drinking episode; thus, the effect is not related to changes in the pharmacokinetics of ethanol. The present study shows that HPE markedly reduces ethanol intake in msP rats, without significantly modifying food intake. Publication Types: Research Support, Non-U.S. Gov't PMID: 10528811 [PubMed - indexed for MEDLINE] 2183: Proc Natl Acad Sci U S A. 1999 Sep 28;96(20):11676-80. Antisense-mediated silencing of a gene encoding a major ryegrass pollen allergen. Bhalla PL, Swoboda I, Singh MB. Plant Molecular Biology and Biotechnology Laboratory, Institute of Land and Food Resources, University of Melbourne, Parkville, Victoria 3052, Australia. p.bhalla@landfood.unimelb.edu.au Type 1 allergic reactions, such as hay fever and allergic asthma, triggered by grass pollen allergens are a global health problem that affects approximately 20% of the population in cool, temperate climates. Ryegrass is the dominant source of allergens because of its prodigious production of airborne pollen. Lol p 5 is the major allergenic protein of ryegrass pollen, judging from the fact that almost all of the individuals allergic to grass pollen show presence of serum IgE antibodies against this protein. Moreover, nearly two-thirds of the IgE reactivity of ryegrass pollen has been attributed to this protein. Therefore, it can be expected that down-regulation of Lol p 5 production can significantly reduce the allergic potential of ryegrass pollen. Here, we report down-regulation of Lol p 5 with an antisense construct targeted to the Lol p 5 gene in ryegrass. The expression of antisense RNA was regulated by a pollen-specific promoter. Immunoblot analysis of proteins with allergen-specific antibodies did not detect Lol p 5 in the transgenic pollen. The transgenic pollen showed remarkably reduced allergenicity as reflected by low IgE-binding capacity of pollen extract as compared with that of control pollen. The transgenic ryegrass plants in which Lol p 5 gene expression is perturbed showed normal fertile pollen development, indicating that genetic engineering of hypoallergenic grass plants is possible. Publication Types: Research Support, Non-U.S. Gov't PMID: 10500236 [PubMed - indexed for MEDLINE] 2184: Science. 1999 Sep 3;285(5433):1491; author reply 1492. Comment on: Science. 1999 Jul 16;285(5426):335. Redesigning evolution? Nader C, Herbert MR, Billings PR, Bereano PL, Hubbard R, King J, Krimsky S, Newman SA, Stabinsky D. Publication Types: Comment Letter PMID: 10498533 [PubMed - indexed for MEDLINE] 2185: Science. 1999 Sep 3;285(5433):1489-91; author reply 1492. Comment on: Science. 1999 Jul 16;285(5426):335. Redesigning evolution? Lovins AB. Publication Types: Comment Letter PMID: 10498532 [PubMed - indexed for MEDLINE] 2186: J AOAC Int. 1999 Jul-Aug;82(4):923-8. IUPAC collaborative trial study of a method to detect genetically modified soy beans and maize in dried powder. Lipp M, Brodmann P, Pietsch K, Pauwels J, Anklam E, Börchers T, Braunschweiger G, Busch U, Eklund E, Eriksen FD, Fagan J, Fellinger A, Gaugitsch H, Hayes D, Hertel C, Hörtner H, Joudrier P, Kruse L, Meyer R, Miraglia M, Müller W, Phillipp P, Pöpping B, Rentsch R, Wurtz A, et al. Institute for Health and consumer protection, Food Products unit, Ispra(Va), Italy. This paper presents results of a collaborative trial study (IUPAC project No. 650/93/97) involving 29 laboratories in 13 countries applying a method for detecting genetically modified organisms (GMOs) in food. The method is based on using the polymerase chain reaction to determine the 35S promotor and the NOS terminator for detection of GMOs. reference materials were produced that were derived from genetically modified soy beans and maize. Correct identification of samples containing 2% GMOs is achievable for both soy beans and maize. For samples containing 0.5% genetically modified soy beans, analysis of the 35S promotor resulted also in a 100% correct classification. However, 3 false-negative results (out of 105 samples analyzed) were reported for analysis of the NOS terminator, which is due to the lower sensitivity of this method. Because of the bigger genomic DNA of maize, the probability of encountering false-negative results for samples containing 0.5% GMOs is greater for maize than for soy beans. For blank samples (0% GMO), only 2 false-positive results for soy beans and one for maize were reported. These results appeared as very weak signals and were most probably due to contamination of laboratory equipment. PMID: 10490320 [PubMed - indexed for MEDLINE] 2187: Int J Food Microbiol. 1999 Sep 15;50(1-2):25-31. The safety and social acceptance of novel foods. Moseley BE. The regulatory processes employed in the UK and the European Union to assess the safety of novel foods and novel food ingredients, including those resulting from the application of recombinant DNA technology (genetically modified foods), are described. Examples are given of yeasts that have been genetically modified and can be used in food and drink manufacture and food enzymes derived from genetically modified microorganisms that have been deemed safe for use by the UK regulatory system. Social acceptance of such novel foods or food ingredients is not uniform in countries of the developed world. Consumer concerns can be based on ethical considerations (scientists "playing God") or safety worries ("more testing needs to be done"). The general acceptance of such foods and food ingredients in Europe is still unclear. PMID: 10488841 [PubMed - indexed for MEDLINE] 2188: Science. 1999 Aug 20;285(5431):1221-2. Comment on: Science. 1999 Aug 20;285(5431):1236-41. Xenografts and retroviruses. Weiss RA. Wohl Virion Centre, Windeyer Institute of Medical Sciences, Univesity College, London W1) 6DB, UK. r.weiss@ucl.ac.uk Publication Types: Comment PMID: 10484730 [PubMed - indexed for MEDLINE] 2189: Lancet. 1999 Aug 21;354(9179):684. Comment on: Lancet. 1999 May 29;353(9167):1811. Health risks of genetically modified foods. Ewen SW, Pusztai A. Publication Types: Comment Letter PMID: 10466701 [PubMed - indexed for MEDLINE] 2190: Transgenic Res. 1998 May;7(3):157-63. Biosafety of E. coli beta-glucuronidase (GUS) in plants. Gilissen LJ, Metz PL, Stiekema WJ, Nap JP. Department of Molecular Biology, CPRO-DLO, Wageningen, The Netherlands. The beta-glucuronidase (GUS) gene is to date the most frequently used reporter gene in plants. Marketing of crops containing this gene requires prior evaluation of their biosafety. To aid such evaluations of the GUS gene, irrespective of the plant into which the gene has been introduced, the ecological and toxicological aspects of the gene and gene product have been examined. GUS activity is found in many bacterial species, is common in all tissues of vertebrates and is also present in organisms of various invertebrate taxa. The transgenic GUS originates from the enterobacterial species Escherichia coli that is widespread in the vertebrate intestine, and in soil and water ecosystems. Any GUS activity added to the ecosystem through genetically modified plants will be of no or minor influence. Selective advantages to genetically modified plants that posses and express the E. coli GUS transgene are unlikely. No increase of weediness of E. coli GUS expressing crop plants, or wild relatives that might have received the transgene through outcrossing, is expected. Since E. coli GUS naturally occurs ubiquitously in the digestive tract of consumers, its presence in food and feed from genetically modified plants is unlikely to cause any harm. E. coli GUS in genetically modified plants and their products can be regarded as safe for the environment and consumers. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 10461390 [PubMed - indexed for MEDLINE] 2191: Toxicology. 1999 Feb 15;132(2-3):99-110. In vitro screening of food peptides toxic for coeliac and other gluten-sensitive patients: a review. Silano M, De Vincenzi M. III Scuola di Specializzazione in Pediatria, IV Clinica Pediatrica, Ospedale S. Paolo, Università di Milano, Milan, Italy. Experience gained through investigations on coeliac disease makes it possible to propose a screening method based on agglutination of isolated K562(S) cells to evaluate the occurrence in food protein of amino acid sequences that are able to adversely affect coeliac and related gluten-sensitive patients. The method consists of in vitro sequential peptic and tryptic digestion of food protein fractions under optimal pH, temperature and time conditions and in vitro incubation of the digest with K562(S) cells; the toxic potential is detected as an agglutination of K 562 (S) cells after a short incubation. Other in vitro test systems, including atrophic coeliac intestinal mucosa and rat fetal intestine, can be used to confirm the results obtained with the isolated cells. A fractionation step of the proteolytic digest on a sepharose-mannan column before exposure of the in vitro systems to the separated peptide fractions adds to the sensitivity of the method. This screening method is not only very useful to investigate action mechanisms in coeliac disease, but also to assess the safety of genetically-modified plant foods and novel foods for gluten-sensitive patients. Publication Types: Review PMID: 10433373 [PubMed - indexed for MEDLINE] 2192: Nature. 1999 Jul 22;400(6742):375-8. Circadian rhythms in olfactory responses of Drosophila melanogaster. Krishnan B, Dryer SE, Hardin PE. Department of Biology and Biochemistry and Biological Clocks Program, University of Houston, Texas 77204-5513, USA. The core mechanism of circadian timekeeping in arthropods and vertebrates consists of feedback loops involving several clock genes, including period (per) and timeless (tim). In the fruitfly Drosophila, circadian oscillations in per expression occur in chemosensory cells of the antennae, even when the antennae are excised and maintained in isolated organ culture. Here we demonstrate a robust circadian rhythm in Drosophila in electrophysiological responses to two classes of olfactory stimuli. These rhythms are observed in wild-type flies during light-dark cycles and in constant darkness, but are abolished in per or tim null-mutant flies (per01 and tim01) which lack rhythms in adult emergence and locomotor behaviour. Olfactory rhythms are also abolished in the per 7.2:2 transgenic line in which per expression is restricted to the lateral neurons of the optic lobe. Because per 7.2:2 flies do not express per in peripheral oscillators, our results provide evidence that peripheral circadian oscillators are necessary for circadian rhythms in olfactory responses. As olfaction is essential for food acquisition, social interactions and predator avoidance in many animals, circadian regulation of olfactory systems could have profound effects on the behaviour of organisms that rely on this sensory modality. Publication Types: Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. PMID: 10432117 [PubMed - indexed for MEDLINE] 2193: Science. 1999 Jul 16;285(5426):387-9. Biotechnology and food security in the 21st century. Serageldin I. Consultative Group on International Agricultural Research, World Bank, 1818 H Street, NW, Washington, DC 20433, USA. Biotechnology can contribute to future food security if it benefits sustainable small-farm agriculture in developing countries. Presently, agrobiotechnology research cites ethical, safety, and intellectual property rights issues. Protection of intellectual property rights encourages private sector investment in agrobiotechnology, but in developing countries the needs of smallholder farmers and environmental conservation are unlikely to attract private funds. Public investment will be needed, and new and imaginative public-private collaboration can make the gene revolution beneficial to developing countries. This is crucial for the well-being of today's hungry people and future generations. PMID: 10411497 [PubMed - indexed for MEDLINE] 2194: Science. 1999 Jul 16;285(5426):384-7. Retraction in: Gaskell G, Bauer M, Durant J, Allum N. Science. 2000 Jun 9;288(5472):1751. Worlds apart? The reception of genetically modified foods in Europe and the U.S. Gaskell G, Bauer MW, Durant J, Allum NC. Methodology Institute, Department of Social Psychology, London School of Economics, London WC2A 2AE, UK. G.Gaskell@lse.ac.uk Recent controversies about genetically modified foods in the United Kingdom and several other European countries highlight the apparent differences that exist in public opinion on this subject across the Atlantic. Why are people in the United States seemingly untroubled by a technology that causes Europeans so many difficulties? The results of survey research on public perceptions of biotechnology in Europe and the United States during 1996-1997, together with an analysis of press coverage and policy formation from 1984 to 1996, can help to answer this question. Publication Types: Research Support, Non-U.S. Gov't Retracted Publication PMID: 10411496 [PubMed - indexed for MEDLINE] 2195: Science. 1999 Jul 16;285(5426):372-5. Gene discovery and product development for grain quality traits. Mazur B, Krebbers E, Tingey S. DuPont Agricultural Products Experimental Station, Post Office Box 80402, Wilmington, DE 19880-0402, USA. The composition of oils, proteins, and carbohydrates in seeds of corn, soybean, and other crops has been modified to produce grains with enhanced value. Both plant breeding and molecular technologies have been used to produce plants carrying the desired traits. Genomics-based strategies for gene discovery, coupled with high-throughput transformation processes and miniaturized, automated analytical and functionality assays, have accelerated the identification of product candidates. Molecular marker-based breeding strategies have been used to accelerate the process of moving trait genes into high-yielding germplasm for commercialization. These products are being tested for applications in food, feed, and industrial markets. Publication Types: Review PMID: 10411493 [PubMed - indexed for MEDLINE] 2196: Nat Biotechnol. 1999 Jul;17(7):631-2. EuropaBio unit created to boost agbio defense. Dorey E. Publication Types: News PMID: 10409346 [PubMed - indexed for MEDLINE] 2197: Nat Biotechnol. 1999 Jul;17(7):628. US agbio controversies continue. Fox JL. Publication Types: News PMID: 10409343 [PubMed - indexed for MEDLINE] 2198: Lancet. 1999 Jul 3;354(9172):71. Comment in: Lancet. 1999 Aug 21;354(9179):684. Comment on: Lancet. 1999 May 22;353(9166):1769. Health risks of genetically modified foods. Brunner E, Millstone E. Publication Types: Comment Letter PMID: 10406385 [PubMed - indexed for MEDLINE] 2199: Lancet. 1999 Jul 3;354(9172):70-1. Comment on: Lancet. 1999 May 29;353(9167):1811. Health risks of genetically modified foods. Schellekens H. Publication Types: Comment Letter PMID: 10406384 [PubMed - indexed for MEDLINE] 2200: Lancet. 1999 Jul 3;354(9172):70. Comment on: Lancet. 1999 May 29;353(9167):1811. Health risks of genetically modified foods. Feldbaum CB. Publication Types: Comment Letter PMID: 10406383 [PubMed - indexed for MEDLINE] 2201: Lancet. 1999 Jul 3;354(9172):69-70. Comment on: Lancet. 1999 May 29;353(9167):1811. Health risks of genetically modified foods. Malcom AD. Publication Types: Comment Letter PMID: 10406382 [PubMed - indexed for MEDLINE] 2202: Lancet. 1999 Jul 3;354(9172):69. Comment on: Lancet. 1999 May 29;353(9167):1811. Health risks of genetically modified foods. Lachmann P. Publication Types: Comment Letter PMID: 10406381 [PubMed - indexed for MEDLINE] 2203: Nature. 1999 Jul 1;400(6739):14. Comment on: Nature. 1999 Apr 22;398(6729):651-6. Making sense of GM tomatoes. Genetically modified. Bright S, Schuch W. Publication Types: Comment Letter PMID: 10403240 [PubMed - indexed for MEDLINE] 2204: Nahrung. 1999 Jun;43(3):168-74. Assessment of allergic potential of (novel) foods. Wal JM. INRA-CEA/SPI, Laboratoire d'Immuno-Allergie Alimentaire, Gif sur Yvette, France. In safety assessment of Novel Foods such as functional foods, allergy is a special issue on which particular emphasis has been placed. The reason for such concern is that incidence of food allergies is constantly and rapidly increasing. The severity of the reported incidents and the number of foods incriminated are also on the rise. The outstanding challenge is to understand what makes a common innocuous protein or peptide behave as an allergen for some groups of people, or why it may suddenly or progressively become a much more potent allergen than usual. It is therefore necessary to consider the risks of creating or unmasking new immunoreactive structures, or of overexposure to already reactive substances, as a result of new food-production and processing technologies. No test such as the use of animal models, the analysis of structure, function and physico-chemical properties is as yet available to evaluate or predict the allergenicity of a "novel" protein in a wholly reliable and objective manner. No indication has yet suggested that novel foods, and particularly recombinant proteins or genetically modified foods, would be more (or less) allergenic than the corresponding conventional foods. No particular structure can be described as being solely and intrinsically allergenic. The predictive approaches to determining the allergenic potential of NFs should therefore be subject to case-by-case critical appraisal allied to mandatory implementation of monitoring of the potential postmarketing impact of these new foodstuffs on public health. Publication Types: Review PMID: 10399350 [PubMed - indexed for MEDLINE] 2205: Int Arch Allergy Immunol. 1999 Jun;119(2):75-85. Genetically engineered plant allergens with reduced anaphylactic activity. Singh MB, de Weerd N, Bhalla PL. Plant Molecular Biology and Biotechnology Laboratory, Institute of Land and Food Resources, University of Melbourne, Parkville, Vic., Australia. Allergy immunotherapy is based on the administration of increasing amounts of the disease-eliciting allergens in order to yield allergen-specific non-responsiveness. Success of this therapy is associated with modulation of the immune response to allergenic molecules at the level of T-helper cells and the induction of blocking antibodies. The extracts used for immunotherapy are highly heterogenous preparations from natural sources and contain additional components, mostly proteins which are not well defined. Recombinant DNA technology offers novel tools for production of pure and well-characterised allergens for specific immunotherapy. However, high IgE reactivity of pure recombinant allergens is associated with an increased risk of potentially life-threatening anaphylactic reactions. A major improvement in allergen-specific immunotherapy may be achieved by using genetically engineered recombinant allergens with reduced anaphylactic activity. Recently the site- directed mutagenesis technique has been applied successfully to produce variants of major grass, birch and oilseed rape allergens with reduced IgE reactivity but retained T-cell reactivity. These modified allergens with reduced anaphylactic potential are novel candidates for safer and more effective allergen-specific immunotherapy. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 10394098 [PubMed - indexed for MEDLINE] 2206: Regul Toxicol Pharmacol. 1999 Jun;29(3):327-57. A cancer risk assessment of di(2-ethylhexyl)phthalate: application of the new U.S. EPA Risk Assessment Guidelines. Doull J, Cattley R, Elcombe C, Lake BG, Swenberg J, Wilkinson C, Williams G, van Gemert M. University of Kansas Medical Center, Kansas City, Kansas, USA. The current United States Environmental Protection Agency (EPA) classification of di(2-ethylhexyl)phthalate (DEHP) as a B2 "probable human" carcinogen is based on outdated information. New toxicology data and a considerable amount of new mechanistic evidence were used to reconsider the cancer classification of DEHP under EPA's proposed new cancer risk assessment guidelines. The total weight-of-evidence clearly indicates that DEHP is not genotoxic. In vivo administration of DEHP to rats and mice results in peroxisome proliferation in the liver, and there is strong evidence and scientific consensus that, in rodents, peroxisome proliferation is directly associated with the onset of liver cancer. Peroxisome proliferation is a transcription-mediated process that involves activation by the peroxisome proliferator of a nuclear receptor in rodent liver called the peroxisome proliferator-activated receptor (PPARalpha). The critical role of PPARalpha in peroxisomal proliferation and carcinogenicity in mice is clearly established by the lack of either response in mice genetically modified to remove the PPARalpha. Several mechanisms have been proposed to explain how, in rodents, peroxisome proliferation can lead to the formation of hepatocellular tumors. The general consensus of scientific opinion is that PPARalpha-induced mitogenesis and cell proliferation are probably the major mechanisms responsible for peroxisome proliferator-induced hepatocarcinogenesis in rodents. Oxidative stress appears to play a significant role in this increased cell proliferation. It triggers the release of TNFalpha by Kupffer cells, which in turn acts as a potent mitogen in hepatocytes. Rats and mice are uniquely responsive to the morphological, biochemical, and chronic carcinogenic effects of peroxisome proliferators, while guinea pigs, dogs, nonhuman primates, and humans are essentially nonresponsive or refractory; Syrian hamsters exhibit intermediate responsiveness. These differences are explained, in part, by marked interspecies variations in the expression of PPARalpha, with levels of expression in humans being only 1-10% of the levels found in rat and mouse liver. Recent studies of DEHP clearly indicate a nonlinear dose-response curve that strongly suggests the existence of a dose threshold below which tumors in rodents are not induced. Thus, the hepatocarcinogenic effects of DEHP in rodents result directly from the receptor-mediated, threshold-based mechanism of peroxisome proliferation, a well-understood process associated uniquely with rodents. Since humans are quite refractory to peroxisomal proliferation, even following exposure to potent proliferators such as hypolipidemic drugs, it is concluded that the hepatocarcinogenic response of rodents to DEHP is not relevant to human cancer risk at any anticipated exposure level. DEHP should be classified an unlikely human carcinogen with a margin of exposure (MOE) approach to risk assessment. The most appropriate and conservative point of reference for assessing MOEs should be 20 mg/kg/day, which is the mouse NOEL for peroxisome proliferation and increased liver weight. Exposure of the general human population to DEHP is approximately 30 microg/kg body wt/day, the major source being from residues in food. Higher exposures occur occupationally [up to about 700 microg/kg body wt/day (mainly by inhalation) based on current workplace standards] and through use of certain medical devices [e.g., up to 457 microg/kg body wt/day for hemodialysis patients (intravenous)], although these have little relevance because the routes of exposure bypass critical activation enzymes in the gastrointestinal tract. Copyright 1999 Academic Press. Publication Types: Review PMID: 10388618 [PubMed - indexed for MEDLINE] 2207: Endocrinology. 1999 Jul;140(7):3183-7. Selective dependence of intracerebroventricular neuropeptide Y-elicited effects on central glucocorticoids. Zakrzewska KE, Sainsbury A, Cusin I, Rouru J, Jeanrenaud B, Rohner-Jeanrenaud F. Laboratoires de Recherches Métaboliques, Geneva University, School of Medicine, Switzerland. katerina_z@hotmail.com It has been reported that hyperphagia and excessive body weight gain of genetically obese rodents were abolished by adrenalectomy. High hypothalamic levels of neuropeptide Y (NPY) were found in obese rodents. A chronic intracerebroventricular (icv) infusion of NPY in normal rats was shown to produce most hormono-metabolic abnormalities of genetically obese animals, and to be inefficient in doing so in adrenalectomized (ADX) rats. The combined presence of NPY and of glucocorticoids thus appeared to be necessary for inducing obesity. This study, therefore, was aimed at determining the consequences of a chronic i.c.v. NPY infusion in ADX rats receiving or not i.c.v. glucocorticoids. It was found that the combined i.c.v. infusion of NPY and dexamethasone in ADX rats increased food intake, body weight, plasma insulin, leptin, and triglyceride levels relative to vehicle-infused ADX controls. The infusion of NPY alone, or of dexamethasone alone in ADX rats failed to produce these effects. In contrast, the icv infusion of NPY alone greatly decreased the expression of brown adipose tissue uncoupling protein-1 and -3. This was not modified by the superimposed infusion of dexamethasone. It is concluded that, although many of centrally elicited NPY effects require the central presence of glucocorticoids, those bearing on the inhibition of uncoupling proteins expression (energy dissipation) do not require central glucocorticoids. Publication Types: Research Support, Non-U.S. Gov't PMID: 10385413 [PubMed - indexed for MEDLINE] 2208: Science. 1999 May 28;284(5419):1471-2. A rational approach to labeling biotech-derived foods. Miller HI. Hoover Institution, Stanford University, Stanford, CA 94305, USA. miller@hoover.stanford.edu PMID: 10383324 [PubMed - indexed for MEDLINE] 2209: Science. 1999 May 28;284(5419):1442-4. Genetically modified food. Britain struggles to turn anti-GM tide. Gavaghan H. Publication Types: News PMID: 10383316 [PubMed - indexed for MEDLINE] 2210: Curr Probl Dermatol. 1999;28:81-7. Genetically modified food: a danger or a benefit for atopics? Ebner C. Institute of General and Experimental Pathology, University of Vienna, Austria. Publication Types: Review PMID: 10374054 [PubMed - indexed for MEDLINE] 2211: BMJ. 1999 Jun 19;318(7199):1694-5. Comment on: BMJ. 1999 Feb 27;318(7183):547-8. The paradoxes of genetically modified foods. Summary of electronic responses. [No authors listed] Publication Types: Comment Letter PMID: 10438210 [PubMed - indexed for MEDLINE] 2212: BMJ. 1999 Jun 19;318(7199):1694. Comment on: BMJ. 1999 Feb 27;318(7183):547-8. The paradoxes of genetically modified foods. Protection of the public health should underpin all decisions. Plugge E. Publication Types: Comment Letter PMID: 10373182 [PubMed - indexed for MEDLINE] 2213: Curr Opin Biotechnol. 1999 Jun;10(3):298-302. Regulation of foods derived from genetically engineered crops. Mitten DH, MacDonald R, Klonus D. AgrEvo USA, 414 Fourth Street, Suite A, Woodland, CA 95695, USA. Publication Types: Review PMID: 10361083 [PubMed - indexed for MEDLINE] 2214: Lancet. 1999 May 29;353(9167):1811. Comment in: Lancet. 1999 Aug 21;354(9179):684. Lancet. 1999 Jul 3;354(9172):69. Lancet. 1999 Jul 3;354(9172):69-70. Lancet. 1999 Jul 3;354(9172):70. Lancet. 1999 Jul 3;354(9172):70-1. Lancet. 1999 Nov 13;354(9191):1729. Health risks of genetically modified foods. [No authors listed] Publication Types: Editorial PMID: 10359398 [PubMed - indexed for MEDLINE] 2215: Curr Opin Plant Biol. 1999 Apr;2(2):121-2. Comment on: Curr Opin Plant Biol. 1999 Apr;2(2):123-7. Curr Opin Plant Biol. 1999 Apr;2(2):128-34. Curr Opin Plant Biol. 1999 Apr;2(2):135-8. Curr Opin Plant Biol. 1999 Apr;2(2):139-44. Plant metabolic engineering: are we ready for phase two? Ohlrogge J. Publication Types: Comment Editorial PMID: 10357608 [PubMed - indexed for MEDLINE] 2216: BMJ. 1999 May 29;318(7196):1441. Chief medical officer clears genetically modified foods. Beecham L. Publication Types: News PMID: 10346764 [PubMed - indexed for MEDLINE] 2217: Transgenic Res. 1998 Nov;7(6):437-47. Stable production of human insulin-like growth factor 1 (IGF-1) in the milk of hemi- and homozygous transgenic rabbits over several generations. Zinovieva N, Lassnig C, Schams D, Besenfelder U, Wolf E, Müller S, Frenyo L, Seregi J, Müller M, Brem G. Research Center for Milk and Food Weihenstephan, TU Munich, Freising-Weihenstephan. One transgenic rabbit line was generated carrying a fusion gene consisting of the cDNA for human IGF-1 fused to a mammary gland specific expression cassette derived from bovine alpha-S1-casein sequences. Transgene expression was shown to be strictly tissue and lactation period specific. The transgenic rabbit line was bred for six generations. All transgenic animals showed stable production of biologically active IGF-1 over the generations and no apparent effect on the physiological or reproductive performance was observed. The absence of adverse effects on homozygous transgenic rabbits suggested the absence of insertional mutagenesis. Eight hemizygous transgenic offspring analysed produced on average 363 +/- 12 micrograms/ml (ranging from 223 +/- 61 to 484 +/- 39 micrograms/ml) mature human IGF-1 in their milk, whereas three homozygous animals produced on average 543 +/- 41 micrograms/ml (ranging from 360 +/- 15 to 678 +/- 80 micrograms/ml). Homozygous hulGF-1 females clearly showed a significantly increased production performance of the recombinant protein. PMID: 10341452 [PubMed - indexed for MEDLINE] 2218: Proc Natl Acad Sci U S A. 1999 May 25;96(11):5978-81. Transgenic plants for tropical regions: some considerations about their development and their transfer to the small farmer. Herrera-Estrella L. Departamento de Ingeniería Genética, Centro de Investigación y Estudios Avanzados, Apartado Postal 629 C.P. 36500 Irapuato, Guanajuato, Mexico. 1herrera@irapuato.ira.cinvestav.mx Biotechnological applications, especially transgenic plants, probably hold the most promise in augmenting agricultural production in the first decades of the next millennium. However, the application of these technologies to the agriculture of tropical regions where the largest areas of low productivity are located, and where they are most needed, remains a major challenge. In this paper, some of the important issues that need to be considered to ensure that plant biotechnology is effectively transferred to the developing world are discussed. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 10339527 [PubMed - indexed for MEDLINE] 2219: Proc Natl Acad Sci U S A. 1999 May 25;96(11):5973-7. Use of plant roots for phytoremediation and molecular farming. Gleba D, Borisjuk NV, Borisjuk LG, Kneer R, Poulev A, Skarzhinskaya M, Dushenkov S, Logendra S, Gleba YY, Raskin I. Biotech Center, Foran Hall, Cook College, Rutgers University, 59 Dudley Road, New Brunswick, NJ 08901-8520, USA. Alternative agriculture, which expands the uses of plants well beyond food and fiber, is beginning to change plant biology. Two plant-based biotechnologies were recently developed that take advantage of the ability of plant roots to absorb or secrete various substances. They are (i) phytoextraction, the use of plants to remove pollutants from the environment and (ii) rhizosecretion, a subset of molecular farming, designed to produce and secrete valuable natural products and recombinant proteins from roots. Here we discuss recent advances in these technologies and assess their potential in soil remediation, drug discovery, and molecular farming. Publication Types: Review PMID: 10339526 [PubMed - indexed for MEDLINE] 2220: Adv Exp Med Biol. 1999;464:179-94. Autoantigens produced in plants for oral tolerance therapy of autoimmune diseases. Ma S, Jevnikar AM. John P. Robarts Research Institute, University of Western Ontario, London, Canada. Oral administration of protein antigens can induce antigen-specific immune hyporesponsiveness and may be useful in treating autoimmune diseases or preventing transplant rejection. However, the therapeutic value of oral tolerance may be limited when candidate autoantigens cannot be produced by conventional system in quantities sufficient for clinical studies. Plants may be ideally suited for this purpose, as they can produce hugh quantities of functional mammalian proteins at extremely competitive cost. Furthermore, transgenic food plants could provide a simple and direct method of autoantigen delivery for oral tolerance. Here we show that the diabetes-associated autoantigen glutamic acid decarboxylase (GAD) is efficiently expressed in both tobacco and potato plants, and that mice, when fed with fresh transgenic potato tubers, are fully protected from diabetes. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 10335394 [PubMed - indexed for MEDLINE] 2221: Adv Exp Med Biol. 1999;464:161-78. Food plant-delivered cholera toxin B subunit for vaccination and immunotolerization. Arakawa T, Yu J, Langridge WH. Center for Molecular Biology and Gene Therapy, School of Medicine, Loma Linda University, California 92350, USA. Developments in recombinant DNA technology have enabled molecular biologists to introduce a variety of novel genes into plant species for specific purposes. From crop improvement to vaccine antigen and antibody production, plants are attractive bioreactors for production of recombinant proteins, as their eukaryotic nature often permits appropriate post-translational modification of recombinant proteins to retain native biological activity. The autotrophic growth of plants requires only soil minerals, water, nitrogen, sunlight energy and carbon dioxide for the synthesis of constituent proteins. Furthermore, production of biologically active proteins in food plants provides the advantage of direct delivery through consumption of edible transformed plant tissues. The production of cholera toxin B subunit in potato plants and applications for prevention of infectious and autoimmune disease are explained in this contribution. Publication Types: Review PMID: 10335393 [PubMed - indexed for MEDLINE] 2222: Adv Exp Med Biol. 1999;464:149-59. Improvements in human health through production of human milk proteins in transgenic food plants. Arakawa T, Chong DK, Slattery CW, Langridge WH. Center for Molecular Biology and Gene Therapy, Loma Linda University, California 92350, USA. Plants are particularly suitable bioreactors for the production of proteins, as their eukaryotic nature frequently directs the appropriate post-translational modifications of recombinant proteins to retain native biological activity. The autotrophic growth of plants makes this in vivo biosynthesis system economically competitive for supplementation or replacement of conventional production systems in the future. For the production of biologically active proteins, food plants provide the advantage of direct delivery via consumption of transformed plant tissues. Here we describe the production of recombinant human milk proteins in food plants for improvements in human nutrition and health, with emphasis on enhanced nutrition for non-breast fed infants as well as children and adults. Nutritional improvements in edible plants generated through advancements in recombinant DNA technology are rapidly repositioning the world for enjoyment of a more healthful diet for humans in all age groups. Publication Types: Review PMID: 10335392 [PubMed - indexed for MEDLINE] 2223: Adv Exp Med Biol. 1999;464:127-47. Molecular farming of industrial proteins from transgenic maize. Hood EE, Kusnadi A, Nikolov Z, Howard JA. Department of Food Science and Human Nutrition, Iowa State University, Ames 50101, USA. eha105@aol.com Recombinant egg white avidin and bacterial B-glucuronidase (GUS) from transgenic maize have been commercially produced. High levels of expression were obtained in seed by employing the ubiquitin promoter from maize. The recombinant proteins had activities that were indistinguishable from their native counterparts. We have illustrated that down-stream activities in the production of these recombinant proteins, such as stabilizing the germplasm and processing for purification, were accomplished without any major obstacles. Avidin (A8706) and GUS (G2035) are currently marketed by Sigma Chemical Co. Publication Types: Review PMID: 10335391 [PubMed - indexed for MEDLINE] 2224: Curr Opin Plant Biol. 1999 Apr;2(2):135-8. Comment in: Curr Opin Plant Biol. 1999 Apr;2(2):121-2. Commentary: economic aspects of transgenic crops which produce novel products. Hitz B. DuPont Agricultural Products, PO Box 80402, Wilmington, DE 19880-0402, USA. william.d.hitz@usa.dupont.com PMID: 10322200 [PubMed - indexed for MEDLINE] 2225: Praxis (Bern 1994). 1999 Apr 1;88(14):609-14, 616-8. [Food additives and genetically modified food--a risk for allergic patients?] [Article in German] Wüthrich B. Dermatologische Klinik und Poliklinik, Universitätsspital Zürich. Adverse reactions to food and food additives must be classified according to pathogenic criteria. It is necessary to strictly differentiate between an allergy, triggered by a substance-specific immunological mechanism, and an intolerance, in which no specific immune reaction can be established. In contrast to views expressed in the media, by laymen and patients, adverse reactions to additives are less frequent than is believed. Due to frequently "alternative" methods of examination, an allergy to food additives is often wrongly blamed as the cause of a wide variety of symptoms and illness. Diagnosing an allergy or intolerance to additives normally involves carrying out double-blind, placebo-controlled oral provocation tests with food additives. Allergic reactions to food additives occur particularly against additives which are organic in origin. In principle, it is possible that during the manufacture of genetically modified plants and food, proteins are transferred which potentially create allergies. However, legislation exists both in the USA (Federal Drug Administration, FDA) and in Switzerland (Ordinance on the approval process for GM food, GM food additives and GM accessory agents for processing) which require a careful analysis before a genetically modified product is launched, particularly where foreign genes are introduced. Products containing genetically modified organisms (GMO) as additives must be declared. In addition, the source of the foreign protein must be identified. The "Round-up ready" (RR) soya flour introduced in Switzerland is no different from natural soya flour in terms of its allergenic potential. Genetically modified food can be a blessing for allergic individuals if gene technology were to succeed in removing the allergen (e.g. such possibilities exist for rice). The same caution shown towards genetically modified food might also be advisable for foreign food in our diet. Luckily, the immune system of the digestive tract in healthy people tolerates foreign antigens. Food allergies in adults occur mainly among those allergic to pollen. Publication Types: English Abstract Review PMID: 10321121 [PubMed - indexed for MEDLINE] 2226: Science. 1999 Apr 9;284(5412):261. Public openness. Sarewitz D. Publication Types: Letter PMID: 10232970 [PubMed - indexed for MEDLINE] 2227: Lancet. 1999 May 1;353(9163):1531. Comment on: Lancet. 1999 Feb 20;353(9153):605-6. Genetically modified foods. Crawford MA. Publication Types: Comment Letter PMID: 10232352 [PubMed - indexed for MEDLINE] 2228: BMJ. 1999 May 8;318(7193):1284. Comment on: BMJ. 1999 Apr 17;318(7190):1023-4. BMJ. 1999 Feb 27;318(7183):581-4. Genetically modified foods and the Pusztai affair. Rhodes JM. Publication Types: Comment Letter PMID: 10231269 [PubMed - indexed for MEDLINE] 2229: Food Addit Contam. 1998 Oct;15(7):767-74. Development and application of a selective detection method for genetically modified soy and soy-derived products. Hoef AM, Kok EJ, Bouw E, Kuiper HA, Keijer J. Department of Food Safety and Health, State Institute for Quality Control of Agricultural Products (RIKILT-DLO), Wageningen, The Netherlands. A method has been developed to distinguish between traditional soy beans and transgenic Roundup Ready soy beans, i.e. the glyphosate ('Roundup') resistant soy bean variety developed by Monsanto Company. Glyphosate resistance results from the incorporation of an Agrobacterium-derived 5-enol-pyruvyl-shikimate-3-phosphatesynthase (EPSPS) gene. The detection method developed is based on a nested Polymerase Chain Reaction (PCR) procedure. Ten femtograms of soy bean DNA can be detected, while, starting from whole soy beans, Roundup Ready DNA can be detected at a level of 1 Roundup Ready soy bean in 5000 non-GM soy beans (0.02% Roundup Ready soy bean). The method has been applied to samples of soy bean, soy-meal pellets and soy bean flour, as well as a number of processed complex products such as infant formula based on soy, tofu, tempeh, soy-based desserts, bakery products and complex meat and meat-replacing products. The results obtained are discussed with respect to practical application of the detection method developed. Publication Types: Research Support, Non-U.S. Gov't PMID: 10211183 [PubMed - indexed for MEDLINE] 2230: Curr Opin Biotechnol. 1999 Apr;10(2):203-8. Public reactions and scientific responses to transgenic crops. Dale PJ. John Innes Centre, Colney Lane, Norwich NR4 7UH, UK. phil.dale@bbsrc.ac.uk There is currently intense debate in parts of Europe about the commercial production of transgenic food crops. Information from the press and lobbying groups has not encouraged an informed and balanced consideration of the issues. In marked contrast, there is widespread acceptance of transgenic food crops in North America. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 10209143 [PubMed - indexed for MEDLINE] 2231: Nat Biotechnol. 1999 Apr;17(4):311. Comment in: Nat Biotechnol. 1999 Jun;17(6):517. Genetically modified muddle. [No authors listed] Publication Types: Editorial PMID: 10207857 [PubMed - indexed for MEDLINE] 2232: Chin J Biotechnol. 1998;14(2):75-84. Cloning of 1-aminocyclopropane-1-carboxylate (ACC) synthetase cDNA and the inhibition of fruit ripening by its antisense RNA in transgenic tomato plants. Liu C, Tian Y, Shen Q, Jiang H, Ju R, Yan T, Liu C, Mang K. Institute of Botany, Chinese Academy of Sciences, Beijing, China. A 1.7 kb fragment of ACC synthetase cDNA, one member of the ACC synthetase multigene family, was amplified from total tomato cDNA through a polymerase chain reaction (PCR) and cloned in E. coli. Restriction mapping and sequencing analysis confirmed its fidelity and correctness. The cloned ACC synthetase gene was then inserted into a binary vector pBin437, in an inverted orientation between the CaMV 35S promoter with duplicated enhancers and the Nos 3' transcriptional termination sequence, to construct an expression vector pBACC. Transgenic tomato plants were obtained by A. tumefaciens-mediated transformation of cotyledons. PCR detection and Southern blot analysis confirmed the integration of the antisense ACC synthetase gene in the transformed tomato genome. The results from RT-PCR of RNAs isolated from transgenic tomato leaves confirmed that antisense ACC synthetase RNA was synthesized in these transgenic plants. The amount of ethylene released from transgenic tomato fruits was reduced significantly to about 30% of that released by non-transformed controls. The inhibition effect of antisense RNA on fruit ripening was observed in transgenic plants and their progeny (T1). The shelf life of transgenic tomato fruits was at least 60 days at room temperature without significant change in hardness and color. After 15-20 days of treatment of the transgenic fruits with ethylene, most of them reached the ripe stage. The antisense ACC synthetase gene was inherited as a single gene in the progenies of transgenic tomatoes determined by T1 progeny analysis, consistent with the results of Southern blot analysis. Transgenic homozygotes expressing antisense ACC synthetase RNA showed prolonged shelf life in the T2 progeny. Publication Types: Research Support, Non-U.S. Gov't PMID: 10196631 [PubMed - indexed for MEDLINE] 2233: Biosci Biotechnol Biochem. 1999 Feb;63(2):314-8. Quality and safety evaluation of genetically engineered rice with soybean glycinin: analyses of the grain composition and digestibility of glycinin in transgenic rice. Momma K, Hashimoto W, Ozawa S, Kawai S, Katsube T, Takaiwa F, Kito M, Utsumi S, Murata K. Research Institute for Food Science, Kyoto University, Japan. The composition of nutritionally and physiologically important molecules in transgenic rice with the soybean glycinin gene was determined and compared with that of a non-transgenic control. Except for the levels of protein, amino acids and moisture, no marked differences were found between the two kinds of rice. The protein content of the transgenic rice was about 20% higher than the control (control, 6.5 g/100 g; transgenic, 8.0 g/100 g) with a concomitantly lower moisture content. This increased protein content mainly resulted from the increased glycinin expressed in the transgenic rice, and the protein was susceptible to gastric and intestinal digestion juices. In parallel with the increased protein content, some important amino acids lacking in quantity in normal rice were replenished. Publication Types: Research Support, Non-U.S. Gov't PMID: 10192912 [PubMed - indexed for MEDLINE] 2234: Biotechnol Bioeng. 1998 Oct 5;60(1):44-52. Processing of transgenic corn seed and its effect on the recovery of recombinant beta-glucuronidase. Kusnadi AR, Evangelista RL, Hood EE, Howard JA, Nikolov ZL. Department of Food Science and Human Nutrition, 2312 Food Science Building, Iowa State University, Ames, Iowa 50011, USA. The tools of plant biotechnology that have been developed to improve agronomic traits are now being applied to generate recombinant protein products for the food, feed, and pharmaceutical industry. This study addresses several processing and protein recovery issues that are relevant to utilizing transgenic corn as a protein production system. The gus gene coding for beta-glucuronidase (rGUS) was stably integrated and expressed over four generations. The accumulation level of rGUS reached 0.4% of total extractable protein. Within the kernel, rGUS was preferentially accumulated in the germ even though a constitutive ubiquitin promoter was used to direct gus expression. Fourth-generation transgenic seed was used to investigate the effect of seed processing on the activity and the recovery of rGUS. Transgenic seed containing rGUS could be stored at an ambient temperature for up to two weeks and for at least three months at 10 degrees C without a significant loss of enzyme activity. rGUS exposed to dry heat was more stable in ground than in whole kernels. The enzyme stability was correlated with the moisture loss of the samples during the heating. Transgenic seed was dry-milled, fractionated, and hexane extracted to produce full-fat and defatted germ fractions. The results of the aqueous extraction of rGUS from ground kernels, full-fat germ, and defatted-germ samples revealed that approximately 10 times more rGUS per gram of solids could be extracted from the ground full-fat germ and defatted-germ than from the kernel samples. The extraction of corn oil from ground germ with hot hexane (60 degrees C) did not affect the extractable rGUS activity. rGUS was purified from ground kernels and full-fat germ extracts by ion exchange, hydrophobic interaction, and size exclusion chromatography. Similar purity and yield of rGUS were obtained from both extracts. Biochemical properties of rGUS purified from transgenic corn seed were similar to those of E. coli GUS. Copyright 1998 John Wiley & Sons, Inc. PMID: 10099404 [PubMed - indexed for MEDLINE] 2235: Anim Biotechnol. 1998;9(3):vii, 161-228. Proceedings of the Food Animal Biotechnology (FAB) Center Symposium on Transgenic Animals and Food Production. St. Paul, Minnesota, USA. June 26, 1998. [No authors listed] Publication Types: Congresses Overall PMID: 10094593 [PubMed - indexed for MEDLINE] 2236: BMJ. 1999 Feb 20;318(7182):483. Scientists call for moratorium on genetically modified foods. Christie B. Publication Types: News PMID: 10094530 [PubMed - indexed for MEDLINE] 2237: Science. 1999 Feb 19;283(5405):1094-5. Comment in: Science. 2000 Jun 9;288(5472):1748-9. Preliminary data touch off genetic food fight. Enserink M. Publication Types: News PMID: 10075564 [PubMed - indexed for MEDLINE] 2238: Curr Biol. 1999 Mar 11;9(5):R154. Potato hash. Dixon B. PMID: 10074459 [PubMed - indexed for MEDLINE] 2239: Appl Environ Microbiol. 1999 Mar;65(3):1202-6. A general method for selection of alpha-acetolactate decarboxylase-deficient Lactococcus lactis mutants to improve diacetyl formation. Curic M, Stuer-Lauridsen B, Renault P, Nilsson D. Research and Development, Chr. Hansen A/S, DK-2970 Horsholm, Denmark. The enzyme acetolactate decarboxylase (Ald) plays a key role in the regulation of the alpha-acetolactate pool in both pyruvate catabolism and the biosynthesis of the branched-chain amino acids, isoleucine, leucine, and valine (ILV). This dual role of Ald, due to allosteric activation by leucine, was used as a strategy for the isolation of Ald-deficient mutants of Lactococcus lactis subsp. lactis biovar diacetylactis. Such mutants can be selected as leucine-resistant mutants in ILV- or IV-prototrophic strains. Most dairy lactococcus strains are auxotrophic for the three amino acids. Therefore, the plasmid pMC004 containing the ilv genes (encoding the enzymes involved in the biosynthesis of IV) of L. lactis NCDO2118 was constructed. Introduction of pMC004 into ILV-auxotrophic dairy strains resulted in an isoleucine-prototrophic phenotype. By plating the strains on a chemically defined medium supplemented with leucine but not valine and isoleucine, spontaneous leucine-resistant mutants were obtained. These mutants were screened by Western blotting with Ald-specific antibodies for the presence of Ald. Selected mutants lacking Ald were subsequently cured of pMC004. Except for a defect in the expression of Ald, the resulting strain, MC010, was identical to the wild-type strain, as shown by Southern blotting and DNA fingerprinting. The mutation resulting in the lack of Ald in MC010 occurred spontaneously, and the strain does not contain foreign DNA; thus, it can be regarded as food grade. Nevertheless, its application in dairy products depends on the regulation of genetically modified organisms. These results establish a strategy to select spontaneous Ald-deficient mutants from transformable L. lactis strains. Publication Types: Research Support, Non-U.S. Gov't PMID: 10049884 [PubMed - indexed for MEDLINE] 2240: BMJ. 1999 Feb 27;318(7183):581-4. Comment in: BMJ. 1999 May 8;318(7193):1284. Science, medicine, and the future. Genetically modified foods. Jones L. Campden and Chorleywood Food Research Association (CCFRA), Chipping Campden GL55 6LD. Leighton@campden.co.uk Publication Types: Review PMID: 10037638 [PubMed - indexed for MEDLINE] 2241: BMJ. 1999 Feb 27;318(7183):547-8. Comment in: BMJ. 1999 Jun 19;318(7199):1694. BMJ. 1999 Jun 19;318(7199):1694-5. The paradoxes of genetically modified foods. Dixon B. Publication Types: Editorial PMID: 10037608 [PubMed - indexed for MEDLINE] 2242: Lancet. 1999 Feb 20;353(9153):605-6. Comment in: Lancet. 1999 May 1;353(9163):1531. Keeping watch over genetically modified crops and foods. Beringer J. School of Biological Sciences, University of Bristol, UK. PMID: 10030321 [PubMed - indexed for MEDLINE] 2243: Rocz Panstw Zakl Hig. 1998;49(3):253-63. [Genetically modified organisms (GMO): toxicological aspects] [Article in Polish] Ludwicki JK. Zakład Toksykologii Srodowiskowej, Państwowy Zakład Higieny, Warszawa. The genetically modified organisms (GMO) are one of the major public concerns partially due to the activity of the non-governmental organizations which believe that public opinion must be duly informed on what leaves the laboratories and enters the environment or is proposed as food. This article discusses some major toxicological and nutritional aspects of GMO designed as food for humans. The range of current use of GMOs, potential hazards for humans, safety assessment, allergenic concerns, and some aspects of the use of marker genes are discussed in regard to human safety. The need for relevant regulations is stressed. Publication Types: English Abstract PMID: 9930018 [PubMed - indexed for MEDLINE] 2244: Transgenic Res. 1998 Sep;7(5):379-86. Novel and transgenic food crops: overview of scientific versus public perception. Ruibal-Mendieta NL, Lints FA. European Interuniversity Association on Society, Science and Technology (ESST), Université Catholique de Louvain. Recombinant DNA technology offers opportunities to develop new products in many different fields, including agriculture and the agro-food area. Transgenic plants with improved agronomic traits currently grow in field trials and a few varieties have already reached the European market. By and large, new technologies raise both concerns and expectations and modern biotechnology is no exception. Indeed, a voluntary moratorium on experiments involving recombinant DNA molecules was called for in 1974. At the present time, although a majority of academic and industrial scientists agree that transgenic food crops pose no risk for the environment or human health, some others believe that certain applications of modern plant biotechnology are hazardous. In particular, deliberate releases of genetically modified plants are regarded as risky. There is also a disparity between expert and lay perception of r-DNA technology applications to food crops, which makes public information a difficult task. This paper aims at exposing these conflicting points of view on the agricultural applications of modern biotechnology. We also propose some recommendations pertaining to public information in Europe. It appears that consensus conferences might be a good approach to stimulate public information and public debate in Europe, although this approach has to be adapted to the cultural context of each country. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 9859226 [PubMed - indexed for MEDLINE] 2245: Transgenic Res. 1998 Sep;7(5):357-69. Expression of a novel piscine growth hormone gene results in growth enhancement in transgenic tilapia (Oreochromis niloticus). Rahman MA, Mak R, Ayad H, Smith A, Maclean N. Division of Cell Science, School of Biological Sciences, University of Southampton, UK. Several lines of transgenic G1 and G2 tilapia fish (Oreochromis niloticus) have been produced following egg injection with gene constructs carrying growth hormone coding sequences of fish origin. Using a construct in which an ocean pout antifreeze promoter drives a chinook salmon growth hormone gene, dramatic growth enhancement has been demonstrated, in which the mean weight of the 7 month old G2 transgenic fish is more than three fold that of their non transgenic siblings. Somewhat surprisingly G1 fish transgenic for a construct consisting of a sockeye salmon metallothionein promoter spliced to a sockeye salmon growth hormone gene exhibited no growth enhancement, although salmon transgenic for this construct do show greatly enhanced growth. The growth enhanced transgenic lines were also strongly positive in a radio-immuno assay for the specific hormone in their serum, whereas the non growth enhanced lines were negative. Attempts to induce expression from the metallothionein promoter by exposing fish to increased levels of zinc were also unsuccessful. Homozygous transgenic fish have been produced from the ocean pout antifreeze/chinook salmon GH construct and preliminary trials suggest that their growth performance is similar to that of the hemizygous transgenics. No abnormalities were apparent in the growth enhanced fish, although minor changes to skull shape and reduced fertility were noted in some fish. There is also preliminary evidence for improved food conversion ratios when growth enhanced transgenic tilapia are compared to their non-transgenic siblings. The long term objective of this study is to produce lines of tilapia which are both growth enhanced and sterile, so offering improved strains of this important food fish for aquaculture. Publication Types: Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. PMID: 9859224 [PubMed - indexed for MEDLINE] 2246: Indian J Exp Biol. 1998 Aug;36(8):811-5. Effect of hexachlorocyclohexane on hsp26 expression in transgenic Drosophila melanogaster. Chowdhuri DK, Saxena DK, Viswanathan PN. Embryotoxicology Section, Industrial Toxicology Research Centre, Lucknow, India. intox@itrc.sirnetd.ernet.in Hexachlorocyclohexane (HCH) at different concentrations (0.5 to 5.0 ng/ml) mixed with food was fed to third instar larvae of hsp26-lacZ transgenic Drosophila for 2 hr and hsp26 gene expression was examined by beta-galactosidase staining. Puffing in salivary gland polytene chromosomes was also studied. Brain and midgut of larvae showed dark blue staining at 2.0 and 5.0 ng/ml of HCH. Absence of induced puffing at 67B and 84D in the salivary gland polytene chromosomes indicated that these glands are not affected. The study suggests presence of the vulnerable sites in larvae to the higher concentrations of HCH. PMID: 9838884 [PubMed - indexed for MEDLINE] 2247: Lett Appl Microbiol. 1998 Nov;27(5):279-82. Survival and biological activity of heat damaged DNA. Masters CI, Miles CA, Mackey BM. Institute of Food Research, Reading, Earley Gate, UK. The thermal degradation of plasmid pUC18 held at temperatures between 100 and 135 degrees C was examined by measuring the ability of heat-treated plasmid preparations to transform Escherichia coli to ampicillin resistance using electroporation. Substantial protection against loss of transforming ability during heating was provided by concentrations of NaCl between 0.25 and 2.0 mol l-1. For example, the addition of 1.0 mol l-1 NaCl to samples heated at 100 degrees C for 15 min increased transformation frequency about 200-fold compared with samples heated without NaCl. In the presence of 0.5-2.0 mol l-1 NaCl, transforming capacity was not destroyed even by heating at 121 degrees C for 15 min, i.e. after a typical sterilization treatment. These findings may have implications for the safe disposal of genetically modified micro-organisms and recombinant DNA preparations. Publication Types: Research Support, Non-U.S. Gov't PMID: 9830145 [PubMed - indexed for MEDLINE] 2248: Am J Clin Nutr. 1998 Nov;68(5):1123-7. Erratum in: Am J Clin Nutr 1999 Apr;69(4):743. Effect of genetically modified, low-phytic acid maize on absorption of iron from tortillas. Mendoza C, Viteri FE, Lönnerdal B, Young KA, Raboy V, Brown KH. Institute of Nutrition of Central America and Panama, Guatemala City, Guatemala. cmendoza@ucdavis.edu BACKGROUND: Genetically modified, low-phytic acid strains of maize were developed to enhance mineral absorption, but have not been tested previously in humans. OBJECTIVES: We evaluated the mineral and phytic acid contents of a low-phytic acid "flint" maize (LPM, the lpa-1-1 mutant) and its parent, wild-type strain (WTM) and measured iron absorption from tortillas prepared with each type of maize and from a reference dose of ferrous ascorbate. DESIGN: Proximate composition and mineral and phytic acid contents were measured by standard techniques. Iron absorption from tortillas was evaluated by using the extrinsic tag method and was measured as the incorporation of radiolabeled iron into the red blood cells of 14 nonanemic men 2 wk after intake. RESULTS: The phytic acid content of LPM was 3.48 mg/g, approximately 35% of the phytic acid content of WTM; concentrations of macronutrients and most minerals were not significantly different between strains. Iron absorption results were adjusted to 40% absorption of ferrous ascorbate. Iron absorption was 49% greater from LPM (8.2% of intake) than from WTM (5.5% of intake) tortillas (P < 0.001, repeated-measures analysis of variance). CONCLUSION: Consumption of genetically modified, low-phytic acid strains of maize may improve iron absorption in human populations that consume maize-based diets. Publication Types: Comparative Study Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. PMID: 9808232 [PubMed - indexed for MEDLINE] 2249: J Food Prot. 1998 Oct;61(10):1265-8. Removal of Escherichia coli O157:H7 from surface tissues of beef carcasses inoculated with wet and dry manure. Delazari I, Iaria ST, Riemann H, Cliver DO, Jothikumar N. Sadia Concordia S.A. Com. Ind., São Paulo, Brazil. Beef tissues were contaminated with wet and dry manure. The manure was previously inoculated with Escherichia coli O157:H7 GFP, genetically modified with a plasmid encoding a protein that fluoresces green when exposed to long-wave ultraviolet light. After incubation at 37 degrees C for 5 days, the wet manure was spread on the surface of beef tissues at an average E. coli O157:H7 GFP level of 6.62 log CFU/cm2. Dry manure was obtained by subjecting wet manure to natural drying (simulating dry manure adhering to the hides of cattle) and was also applied to the surfaces of beef tissues. The degree of removal of E. coli O157:H7 GFP by washing was compared to the removal of cells of the same strain that had been inoculated as a suspension. The E. coli O157:H7 mixed into feces of cattle adhered more strongly to meat surfaces than that applied as a suspension, complicating the removal by conventional washing procedures. The fate of the bacterium mixed into wet or dry manure was evaluated. An initial decrease of the inoculated population was observed; this was probably an effect of the changed environment represented by the manure. After adaptation, the inoculated bacteria grew in the wet manure; a maximum population was reached in 5 days at 37 degrees C; levels declined with drying. The use of the GFP marker was of great value, since it allowed enumeration of E. coli O157:H7 in the presence of the natural flora of manure. PMID: 9798139 [PubMed - indexed for MEDLINE] 2250: Science. 1998 Oct 30;282(5390):943-6. Comment in: Science. 1998 Oct 30;282(5390):856. Extended life-span and stress resistance in the Drosophila mutant methuselah. Lin YJ, Seroude L, Benzer S. Division of Biology, California Institute of Technology, Pasadena, CA 91125, USA. Toward a genetic dissection of the processes involved in aging, a screen for gene mutations that extend life-span in Drosophila melanogaster was performed. The mutant line methuselah (mth) displayed approximately 35 percent increase in average life-span and enhanced resistance to various forms of stress, including starvation, high temperature, and dietary paraquat, a free-radical generator. The mth gene predicted a protein with homology to several guanosine triphosphate-binding protein-coupled seven-transmembrane domain receptors. Thus, the organism may use signal transduction pathways to modulate stress response and life-span. Publication Types: Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S. PMID: 9794765 [PubMed - indexed for MEDLINE] 2251: Nat Biotechnol. 1998 Oct;16(10):934-8. A plant-based cholera toxin B subunit-insulin fusion protein protects against the development of autoimmune diabetes. Arakawa T, Yu J, Chong DK, Hough J, Engen PC, Langridge WH. Center for Molecular Biology and Gene Therapy, Department of Microbiology and Molecular Genetics, School of Medicine, Loma Linda University, CA 92350, USA. Oral administration of disease-specific autoantigens can prevent or delay the onset of autoimmune disease symptoms. We have generated transgenic potato plants that synthesize human insulin, a major insulin-dependent diabetes mellitus autoantigen, at levels up to 0.05% of total soluble protein. To direct delivery of plant-synthesized insulin to the gut-associated lymphoid tissues, insulin was linked to the C-terminus of the cholera toxin B subunit (CTB). Transgenic potato tubers produced 0.1% of total soluble protein as the pentameric CTB-insulin fusion, which retained GM1-ganglioside binding affinity and native antigenicity of both CTB and insulin. Nonobese diabetic mice fed transformed potato tuber tissues containing microgram amounts of the CTB-insulin fusion protein showed a substantial reduction in pancreatic islet inflammation (insulitis), and a delay in the progression of clinical diabetes. Feeding transgenic potato tissues producing insulin or CTB protein alone did not provide a significant reduction in insulitis or diabetic symptoms. The experimental results indicate that food plants are feasible production and delivery systems for immunotolerization against this T cell-mediated autoimmune disease. Publication Types: Research Support, Non-U.S. Gov't PMID: 9788349 [PubMed - indexed for MEDLINE] 2252: Nat Biotechnol. 1998 Oct;16(10):902-3. Costs drive acceptance of transgenic bioassays. Swan N. Publication Types: News PMID: 9788336 [PubMed - indexed for MEDLINE] 2253: Nat Biotechnol. 1998 Oct;16(10):889. New regulation for labeling genetically modified foods: a solution or a problem? Ramón D, Calvo MD, Peris J. Publication Types: Letter PMID: 9788327 [PubMed - indexed for MEDLINE] 2254: Environ Mol Mutagen. 1998;32(2):106-9. Transgenic animal models for mutagenesis studies: role in mutagenesis research and regulatory testing. MacGregor JT. Center for Drug Evaluation and Research, Food and Drug Administration, Rockville, Maryland 20857, USA. Publication Types: Review PMID: 9776171 [PubMed - indexed for MEDLINE] 2255: Nurs Stand. 1998 Jun 24-30;12(40):18. Fit for a king. Dallard D. PMID: 9739653 [PubMed - indexed for MEDLINE] 2256: Environ Health Perspect. 1998 Sep;106(9):A432-7. Erratum in: Environ Health Perspect 1998 Nov;106(11):A528. Natural born killers. Schmidt CW. This year, 30 million acres of the corn, cotton, and potatoes planted in the United States will have been genetically engineered to produce an endotoxin normally found in the microbe Bacillus thuringiensis (Bt), a self-contained pesticide that will be toxic only to target insects. Transgenic pest-resistant crops are a cost-effective alternative to chemical pesticides, and may offer a way to help feed the world's growing population with minimal environmental impact. PMID: 9721262 [PubMed - indexed for MEDLINE] 2257: Biosci Biotechnol Biochem. 1998 Jul;62(7):1461-4. Safety assessment of genetically engineered food: detection and monitoring of glyphosate-tolerant soybeans. Shirai N, Momma K, Ozawa S, Hashimoto W, Kito M, Utsumi S, Murata K. Research Institute for Food Science, Kyoto University, Japan. A detection technique for the genetically engineered food, glyphosate-tolerant soybean (GTS), was designed. Commercial soybeans imported from North America were cultured in pots and genomic DNA was isolated from their leaves. To detect the genes, promoter and terminator, involved in the expression of glyphosate tolerance, PCR was done using the genomic DNA and chemically synthesized primers specific to the genes. DNAs with predicted sizes were amplified and confirmed by DNA sequencing to be the genes responsible for the expression of glyphosate tolerance. Glyphosate-tolerant soybeans were found to form approximately 1.1% of the commercial soybeans, when commercially available soybeans were cultivated and number of soybeans resistant to glyphosate was found. This level is somewhat lower than an estimated value announced officially on the basis of the cultivation area of the glyphosate-tolerant soybeans. Publication Types: Research Support, Non-U.S. Gov't PMID: 9720233 [PubMed - indexed for MEDLINE] 2258: J Appl Microbiol. 1998 Jun;84(6):969-80. Common DNA sequences with potential for detection of genetically manipulated organisms in food. MacCormick CA, Griffin HG, Underwood HM, Gasson MJ. Institute of Food Research, Colney, UK. caroline.maccormick@bbsrc.ac.uk Foods produced by genetic engineering technology are now appearing on the market and many more are likely to emerge in the future. The safety aspects, regulation, and labelling of these foods are still contentious issues in most countries and recent surveys highlight consumer concerns about the safety and labelling of genetically modified food. In most countries it is necessary to have approval for the use of genetically manipulated organisms (GMOs) in the production of food. In order to police regulations, a technology to detect such foods is desirable. In addition, a requirement to label approved genetically modified food would necessitate a monitoring system. One solution is to 'tag' approved GMOs with some form of biological or genetic marker, permitting the surveillance of foods for the presence of approved products of genetic engineering. While non-approved GMOs would not be detected by such a surveillance, they might be detected by a screen for DNA sequences common to all or most GMOs. This review focuses on the potential of using common DNA sequences as detection probes for GMOs. The identification of vector sequences, plant transcription terminators, and marker genes by PCR and hybridization techniques is discussed. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 9717281 [PubMed - indexed for MEDLINE] 2259: Nat Biotechnol. 1998 Aug;16(8):712-3. Science-free GM good tests advance. Peerenboom E. Publication Types: News PMID: 9702761 [PubMed - indexed for MEDLINE] 2260: Nat Biotechnol. 1998 Aug;16(8):698-9. Comment in: Nat Biotechnol. 1998 Aug;16(8):697-8. A baroque solution to a nonproblem. Miller HI, Huttner SL. Stanford University's Hoover Institution, USA. miller@hoover.stanford.edu PMID: 9702753 [PubMed - indexed for MEDLINE] 2261: Nat Biotechnol. 1998 Aug;16(8):697-8. Comment on: Nat Biotechnol. 1998 Aug;16(8):698-9. Socioeconomics and the protocol on biosafety. Crompton T, Wakeford T. Liaison Office, International Centre for Genetic Engineering and Biotechnology, Vienna. tcrompton@unido.org Publication Types: Comment PMID: 9702752 [PubMed - indexed for MEDLINE] 2262: Mol Cell. 1998 Jun;1(7):1057-64. Comment in: Mol Cell. 1998 Jun;1(7):1065-6. The Drosophila Polycomb group gene pleiohomeotic encodes a DNA binding protein with homology to the transcription factor YY1. Brown JL, Mucci D, Whiteley M, Dirksen ML, Kassis JA. Laboratory of Developmental Biology, Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, Maryland 20892, USA. Genes of the Polycomb group (PcG) of Drosophila encode proteins necessary for the maintenance of transcriptional repression of homeotic genes. PcG proteins are thought to act by binding as multiprotein complexes to DNA through Polycomb group response elements (PREs); however, specific DNA binding has not been demonstrated for any of the PcG proteins. We have identified a sequence-specific DNA binding protein that interacts with a PRE from the Drosophila engrailed gene. This protein (PHO) is a homolog of the ubiquitous mammalian transcription factor Yin Yang-1 and is encoded by pleiohomeotic, a known member of the PcG. We propose that PHO acts to anchor PcG protein complexes to DNA. Publication Types: Research Support, Non-U.S. Gov't PMID: 9651589 [PubMed - indexed for MEDLINE] 2263: Tanpakushitsu Kakusan Koso. 1998 Apr;43(5):634-48. [Molecular mechanism of defense system against photooxidative damage in photosynthetic organisms--possibility of creation of plants with tolerance to photooxidative damage] [Article in Japanese] Shigeoka S, Ishikawa T, Takeda T, Tamoi M. Department of Food and Nutrition, Kinki University, Nara, Japan. Publication Types: Review PMID: 9564781 [PubMed - indexed for MEDLINE] 2264: Mutat Res. 1998 Jun 5;401(1-2):165-78. Comparison of in vivo mutagenesis in the endogenous Hprt gene and the lacI transgene of Big Blue(R) rats treated with 7, 12-dimethylbenz[a]anthracene. Manjanatha MG, Shelton SD, Aidoo A, Lyn-Cook LE, Casciano DA. Department of Health and Human Services, Food and Drug Administration, National Center for Toxicological Research, Division of Genetic Toxicology, Jefferson, AR 72079, USA. mmanjanatha@nctr.fda.gov The lacI transgene of Big Blue(R) (BB) rats was evaluated as a reporter of in vivo mutation by comparing mutant frequencies (MFs) in it and in the endogenous Hprt gene. Seven-week old female BB rats were given single doses of 0, 20, 75 and 130 mg/kg of 7, 12-dimethylbenz(a)anthracene (DMBA) by gavage, and Hprt and lacI MFs in splenic lymphocytes were measured over a period of 18 weeks. The Hprt MFs in treated rats increased for 10 weeks and then declined; 130 mg/kg of DMBA produced a maximum Hprt MF of 168+/-11.4x10-6 clonable lymphocytes, while the MF in control rats was 7.4+/-1. 5x10-6. DMBA exposure of generic F344 rats resulted in a similar time-course of mutant induction but produced about 50% higher Hprt MFs with the 75 and 130 mg/kg doses. In contrast, the lacI MFs increased for 6 weeks and then remained relatively constant; 130 mg/kg of DMBA produced a maximum increase in lacI MF of 341+/-83x10-6 PFU compared with 25+/-5x10-6 PFU in control rats. The Hprt mutant frequencies in DMBA-treated BB and F344 rats were significantly increased over control values for every dose-time combination examined, while only the 130 mg/kg dose consistently produced lacI MFs that were significantly above the controls. In addition, the fold-increase in MF for treated vs. control rats was two times higher for the Hprt gene than the lacI gene due to the higher MFs in the lacI gene of control rats. Differences between the lacI and Hprt genes in the kinetics of mutant induction, in the frequency of induced mutants, and in the sensitivity of mutant detection could be explained at least partially by the properties of these two genes. Copyright 1998 Elsevier Science B.V. All rights reserved. Publication Types: Comparative Study PMID: 9639698 [PubMed - indexed for MEDLINE] 2265: BMJ. 1998 Jun 20;316(7148):1845-6. Comment in: BMJ. 1999 Jan 30;318(7179):332. Why all the fuss about genetically modified food?. Much depends on who benefits. Burke D. Publication Types: Editorial PMID: 9632400 [PubMed - indexed for MEDLINE] 2266: Adv Food Nutr Res. 1998;42:63-92. Sequence databases for assessing the potential allergenicity of proteins used in transgenic foods. Gendel SM. Biotechnology Studies Branch, Food and Drug Administration, National Center for Food Safety and Technology, Summit-Argo, Illinois 60501, USA. Publication Types: Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. Review PMID: 9597725 [PubMed - indexed for MEDLINE] 2267: Adv Food Nutr Res. 1998;42:45-62. The use of amino acid sequence alignments to assess potential allergenicity of proteins used in genetically modified foods. Gendel SM. Biotechnology Studies Branch, Food and Drug Administration, National Center for Food Safety and Technology, Summit-Argo, Illinois 60501, USA. Publication Types: Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. Review PMID: 9597724 [PubMed - indexed for MEDLINE] 2268: Crit Rev Food Sci Nutr. 1998 Apr;38(3):173-258. Textural modification of processing tomatoes. Barrett DM, Garcia E, Wayne JE. Department of Food Science & Technology, University of California, Davis 95616-8598, USA. Knowledge of the textural properties of processing tomatoes is crucial to ensuing product acceptability; measurement, control, and optimization of these properties through judicious selection of varieties and control of unit operations results in products that the consumer prefers. It is important to first define the terms texture, rheology, consistency, and viscosity prior to discussing principles of their measurement. The textural properties of processing tomatoes may be measured using both sensory and objective tests, and the latter may be either destructive or nondestructive in nature. The unique anatomy of tomato fruit (peel, pericarp, columella, and locules) in part dictates the method of texture measurement. Numerous factors, including variety, maturity, genetic modification, cultural particles, and environmental conditions, processing conditions, and calcium addition affect the textural integrity of tomatoes. Textural properties of raw tomatoes and most processed tomato products are reviewed in this article. Publication Types: Review PMID: 9595227 [PubMed - indexed for MEDLINE] 2269: Nat Biotechnol. 1998 May;16(5):401-2. Comment on: Nat Biotechnol. 1997 Dec;15(13):1331-2. Genetic ID. Fagan J. Publication Types: Comment Letter PMID: 9592378 [PubMed - indexed for MEDLINE] 2270: Nat Med. 1998 May;4(5 Suppl):502-3. Pharmaceutical foodstuffs--oral immunization with transgenic plants. Arntzen CJ. Publication Types: News PMID: 9585197 [PubMed - indexed for MEDLINE] 2271: Development. 1998 Jun;125(12):2193-202. Spatial and temporal targeting of gene expression in Drosophila by means of a tetracycline-dependent transactivator system. Bello B, Resendez-Perez D, Gehring WJ. Biozentrum, University of Basel, Klingelbergstrasse 70, CH-4056 Basel, Switzerland. bbello@nimr.mrc.ac.uk In order to evaluate the efficiency of the tetracycline-regulated gene expression system in Drosophila, we have generated transgenic lines expressing a tetracycline-controlled transactivator protein (tTA), with specific expression patterns during embryonic and larval development. These lines were used to direct expression of a tTA-responsive promoter fused to the coding region of either the beta-galactosidase or the homeotic protein Antennapedia (ANTP), under various conditions of tetracycline treatment. We found that expression of beta-galactosidase can be efficiently inhibited in embryos and larvae with tetracycline provided in the food, and that a simple removal of the larvae from tetracycline exposure results in the induction of the enzyme in a time- and concentration-dependent manner. Similar treatments can be used to prevent the lethality associated with the ectopic expression of ANTP in embryos and, subsequently, to control the timing of expression of the homeoprotein ANTP specifically in the antennal imaginal disc. Our results show that the expression of a gene placed under the control of a tetracycline-responsive promoter can be tightly controlled, both spatially by the regulatory sequences driving the expression of tTA and temporally by tetracycline. This provides the basis of a versatile binary system for controlling gene expression in Drosophila, with an additional level of regulation as compared to the general method using the yeast transcription factor GAL4. Publication Types: Research Support, Non-U.S. Gov't PMID: 9584119 [PubMed - indexed for MEDLINE] 2272: Food Addit Contam. 1998 Jan;15(1):1-9. The EC novel foods Regulation--a UK perspective. Tomlinson N. Ministry of Agriculture, Fisheries and Food, London, UK. On 15 May 1997 the EC novel foods Regulation came into effect introducing a statutory pre-market approval system for novel foods across the whole of the European Union. A novel food is defined as a food which has not been consumed to a significant degree and includes foods containing or obtained from genetically modified organisms. The Regulation envisages an initial safety assessment at Member State level, although centralized procedures are available to resolve any objections between Member States. The most controversial aspect of the Regulation relates to the provisions for labelling genetically modified foods. Within the framework of the Regulation there is scope for labelling to be considered on a case by case basis, although the UK is pressing for all foods which may contain genetically modified material to be clearly labelled. To ensure that all Member States follow a consistent approach to the safety assessment of novel foods, the Commission has published a series of guidelines to accompany the regulation. The UK already has a well-developed system for assessing the safety of novel foods dating back to the approval of the first novel food in the UK in 1983. PMID: 9534867 [PubMed - indexed for MEDLINE] 2273: Nat Biotechnol. 1998 Mar;16(3):292-7. Erratum in: Nat Biotechnol 1998 May;16(5):478. Efficacy of a food plant-based oral cholera toxin B subunit vaccine. Arakawa T, Chong DK, Langridge WH. Department of Microbiology and Molecular Genetics, School of Medicine, Loma Linda University, CA 92350, USA. Transgenic potatoes were engineered to synthesize a cholera toxin B subunit (CTB) pentamer with affinity for GMI-ganglioside. Both serum and intestinal CTB-specific antibodies were induced in orally immunized mice. Mucosal antibody titers declined gradually after the last immunization but were restored following an oral booster of transgenic potato. The cytopathic effect of cholera holotoxin (CT) on Vero cells was neutralized by serum from mice immunized with transgenic potato tissues. Following intraileal injection with CT, the plant-immunized mice showed up to a 60% reduction in diarrheal fluid accumulation in the small intestine. Protection against CT was based on inhibition of enterotoxin binding to the cell-surface receptor GMI-ganglioside. These results demonstrate the ability of transgenic food plants to generate protective immunity in mice against a bacterial enterotoxin. Publication Types: Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. PMID: 9528012 [PubMed - indexed for MEDLINE] 2274: Nat Biotechnol. 1998 Mar;16(3):214. Genetically engineered organic food? Hoyle R. PMID: 9527990 [PubMed - indexed for MEDLINE] 2275: Nippon Yakurigaku Zasshi. 1997 Oct;110 Suppl 1:1P-6P. [Transgenic plants as medicine production systems] [Article in Japanese] Okada Y. School of Science and Engineering, Teikyo University, Toyosatodai, Tochigi. Transgenic plants are emerging as an important system for the expression of many recombinant proteins, especially those intended for therapeutic purpose. The production of foreign proteins in plants has several advantages. In terms of required equipment and cost, mass production in plants is far easier to achieve than techniques involving animal cells. Successful production of several proteins in plants, including human serum albumin, haemoglobin, monoclonal antibodies, viral antigens (vaccines), enkephalin, and trichosanthin, has been reported. Particularly, the demonstration that vaccine antigens can be produced in plants in their native, immunogenic forms opens exciting possibilities for the "bio-farming" of vaccines. If the antigens are orally active, food-based "edible vaccines" could allow economical production. In this review, I will discuss the progress that has been made by several groups in what is now an expanding area of medicine research that utilizes transgenic plants. Publication Types: English Abstract Review PMID: 9503395 [PubMed - indexed for MEDLINE] 2276: Allerg Immunol (Paris). 1998 Jan;30(1):9-13. Modifications of allergenicity linked to food technologies. Moneret-Vautrin DA. Service de Médecine D, Hôpital Central, Nancy. The prevalence of food allergies (FA) has increased over the past fifteen years. The reasons suggested are changes in dietary behaviour and the evolution of food technologies. New cases of FA have been described with chayote, rambutan, arguta, pumpkin seeds, custard apple, and with mycoproteins from Fusarium.... Additives using food proteins are at high risk: caseinates, lysozyme, cochineal red, papaïn, alpha-amylase, lactase etc. Heating can reduce allergenicity or create neo-allergens, as well as storage, inducing the synthesis of allergenic stress or PR proteins. Aeroallergens (miles, moulds) contaminate foods and can induce allergic reactions. Involuntary contamination by peanut proteins on production lines is a problem which is not yet solved. Genetically modified plants are at risk of allergenicity, requiring methodological steps of investigations: the comparison of the amino-acid sequence of the transferred protein with the sequence of known allergens, the evaluation of thermo degradability and of the denaturation by pepsin and trypsin are required, as well as the study with sera from patients allergic to the plant producing the gene. The combination of enzymatic hydrolysis, heating, or the development of genetically modified plants may offer new alternatives towards hypoallergenic foods (57 references). Publication Types: Review PMID: 9503097 [PubMed - indexed for MEDLINE] 2277: Biotechnol Prog. 1998 Jan-Feb;14(1):149-55. Production and purification of two recombinant proteins from transgenic corn. Kusnadi AR, Hood EE, Witcher DR, Howard JA, Nikolov ZL. Department of Food Science and Human Nutrition, Iowa State University, Ames 50011, USA. This study reports the production, purification, and characterization of recombinant Escherichia coli beta-glucuronidase (GUS) and chicken egg-white avidin from transgenic corn seed. The avidin and gus genes were stably integrated in the genome and expressed over seven generations. The accumulation levels of avidin and GUS in corn kernel were 5.7% and 0.7% of extractable protein, respectively. Within the kernel, avidin and GUS accumulation was mainly localized to the germ, indicating possible tissue preference of the ubiquitin promoter. The storage-stability studies demonstrated that processed transgenic seed containing GUS or avidin can be stored at 10 degrees C for at least 3 months and at 25 degrees C for up to 2 weeks without a significant loss of activity. The heat-stability experiments indicated that GUS and avidin in the whole kernels were stable at 50 degrees C for up to 1 week. The buffer composition also had an affect on the aqueous extraction of avidin and GUS from ground kernels. Avidin was purified in one step by using 2-iminobiotin agarose, whereas GUS was purified in four steps consisting of adsorption, ion-exchange, hydrophobic interaction, and size-exclusion chromatography. Biochemical properties of purified avidin and GUS were similar to those of the respective native proteins. Publication Types: Research Support, U.S. Gov't, Non-P.H.S. PMID: 9496680 [PubMed - indexed for MEDLINE] 2278: Transgenic Res. 1997 Nov;6(6):403-13. Expression of cholera toxin B subunit oligomers in transgenic potato plants. Arakawa T, Chong DK, Merritt JL, Langridge WH. Department of Microbiology and Molecular Genetics, School of Medicine, Loma Linda University, CA 92350, USA. A gene encoding the cholera toxin B subunit protein (CTB), fused to an endoplasmic reticulum (ER) retention signal (SEKDEL) was inserted adjacent to the bi-directional mannopine synthase P2 promoter in a plant expression vector containing a bacterial luciferase AB fusion gene (luxF) linked to the P1 promoter. Potato leaf explants were transformed by Agrobacterium tumefaciens carrying the vector and kanamycin-resistant plants were regenerated. The CTB-SEKDEL fusion gene was identified in the genomic DNA of bioluminescent plants by polymerase chain reaction amplification. Immunoblot analysis indicated that plant-derived CTB protein was antigenically indistinguishable from bacterial CTB protein, and that oligomeric CTB molecules (M(r) approximately 50 kDa) were the dominant molecular species isolated from transgenic potato leaf and tuber tissues. Similar to bacterial CTB, plant-synthesized CTB dissociated into monomers (M(r) approximately 15 kDa) during heat or acid treatment. The maximum amount of CTB protein detected in auxin-induced transgenic potato leaf and tuber tissues was approximately 0.3% of total soluble plant protein. Enzyme-linked immunosorbent assay methods indicated that plant-synthesized CTB protein bound specifically to GM1-ganglioside, the natural membrane receptor of cholera toxin. In the presence of the SEKDEL signal, CTB protein accumulates in potato tissues and is assembled into an oligomeric form that retains native biochemical and immunological properties. The expression of oligomeric CTB protein with immunological and biochemical properties identical to native CTB protein in edible plants opens the way for preparation of inexpensive food plant-based oral vaccines for protection against cholera and other pathogens in endemic areas throughout the world. Publication Types: Research Support, Non-U.S. Gov't PMID: 9423288 [PubMed - indexed for MEDLINE] 2279: Nat Biotechnol. 1997 Dec;15(13):1332. Rhône-Poulenc rounds up DeKalb and Monsanto. Fox JL. Publication Types: News PMID: 9415877 [PubMed - indexed for MEDLINE] 2280: Nat Biotechnol. 1997 Dec;15(13):1331-2. Comment in: Nat Biotechnol. 1998 May;16(5):401-2. Firms sleuth out transgenic foods. Steinberg D. Publication Types: News PMID: 9415876 [PubMed - indexed for MEDLINE] 2281: Ecotoxicol Environ Saf. 1997 Oct;38(1):63-70. Infectivity and effects of gypsy moth and spruce budworm nuclear polyhedrosis viruses ingested by rainbow trout. Kreutzweiser DP, Ebling PM, Holmes SB. Canadian Forest Service, Great Lakes Forestry Centre, Sault Ste. Marie, Ontario, Canada. Rainbow trout fingerlings were fed dried krill injected with gypsy moth or spruce budworm nuclear polyhedrosis virus (LdNPV and CfNPV, respectively) at a total dose of 1.4 x 10(7) occlusion bodies (OBs) per fish. By the end of the 21-day experimental period there were no adverse effects on fish survival or behavior and no significant differences in feeding rates or growth between treated and control fish. The internal organs of all fish were examined at the end of the experiment and there were no signs of lesions, discoloration, swelling, hemorrhaging, or other aberrations. Visceral tissues were analyzed with a horseradish peroxidase-labeled whole genomic DNA probe (enhanced chemiluminescence procedure) to detect infection by the NPVs. There were no indications of NPV infection (no positive signals) in stomach and intestinal tract tissues of treated fish. High background signals were obtained from liver samples, but further analyses indicated that these were not due to the presence of LdNPV or CfNPV. The protocols outlined here should be applicable to determining infectivity and effects of genetically modified insect viruses on fish. Publication Types: Research Support, Non-U.S. Gov't PMID: 9352215 [PubMed - indexed for MEDLINE] 2282: J Dairy Sci. 1997 Sep;80(9):2213-24. Transgenic dairy cattle: genetic engineering on a large scale. Wall RJ, Kerr DE, Bondioli KR. Gene Evaluation and Mapping Laboratory, USDA-ARS-Livestock and Poultry Science Institute, Beltsville, MD 20705, USA. Amid the explosion of fundamental knowledge generated from transgenic animal models, a small group of scientists has been producing transgenic livestock with goals of improving animal production efficiency and generating new products. The ability to modify mammary-specific genes provides an opportunity to pursue several distinctly different avenues of research. The objective of the emerging gene "pharming" industry is to produce pharmaceuticals for treating human diseases. It is argued that mammary glands are an ideal site for producing complex bioactive proteins that can be cost effectively harvested and purified. Consequently, during the past decade, approximately a dozen companies have been created to capture the US market for pharmaceuticals produced from transgenic bioreactors estimated at $3 billion annually. Several products produced in this way are now in human clinical trials. Another research direction, which has been widely discussed but has received less attention in the laboratory, is genetic engineering of the bovine mammary gland to alter the composition of milk destined for human consumption. Proposals include increasing or altering endogenous proteins, decreasing fat, and altering milk composition to resemble that of human milk. Initial studies using transgenic mice to investigate the feasibility of enhancing manufacturing properties of milk have been encouraging. The potential profitability of gene "pharming" seems clear, as do the benefits of transgenic cows producing milk that has been optimized for food products. To take full advantage of enhanced milk, it may be desirable to restructure the method by which dairy producers are compensated. However, the cost of producing functional transgenic cattle will remain a severe limitation to realizing the potential of transgenic cattle until inefficiencies of transgenic technology are overcome. These inefficiencies include low rates of gene integration, poor embryo survival, and unpredictable transgene behavior. Publication Types: Review PMID: 9313167 [PubMed - indexed for MEDLINE] 2283: Science. 1997 Aug 8;277(5327):834-6. Comment on: Science. 1997 Aug 8;277(5327):763-4. Natural behavior polymorphism due to a cGMP-dependent protein kinase of Drosophila. Osborne KA, Robichon A, Burgess E, Butland S, Shaw RA, Coulthard A, Pereira HS, Greenspan RJ, Sokolowski MB. Department of Biology, York University, 4700 Keele Street, North York, Toronto, Ontario, Canada, M3J 1P3. Naturally occuring polymorphisms in behavior are difficult to map genetically and thus are refractory to molecular characterization. An exception is the foraging gene (for), a gene that has two naturally occurring variants in Drosophila melanogaster food-search behavior: rover and sitter. Molecular mapping placed for mutations in the dg2 gene, which encodes a cyclic guanosine monophosphate (cGMP)-dependent protein kinase (PKG). Rovers had higher PKG activity than sitters, and transgenic sitters expressing a dg2 complementary DNA from rover showed transformation of behavior to rover. Thus, PKG levels affected food-search behavior, and natural variation in PKG activity accounted for a behavioral polymorphism. Publication Types: Comment Research Support, Non-U.S. Gov't PMID: 9242616 [PubMed - indexed for MEDLINE] 2284: Biochem J. 1997 Jul 15;325 ( Pt 2):331-7. Overexpression of arginine decarboxylase in transgenic plants. Burtin D, Michael AJ. Department of Genetics and Microbiology, Institute of Food Research, Norwich Research Park, Colney, Norwich NR47UA, U.K. The activity of arginine decarboxylase (ADC), a key enzyme in plant polyamine biosynthesis, was manipulated in two generations of transgenic tobacco plants. Second-generation transgenic plants overexpressing an oat ADC cDNA contained high levels of oat ADC transcript relative to tobacco ADC, possessed elevated ADC enzyme activity and accumulated 10-20-fold more agmatine, the direct product of ADC. In the presence of high levels of the precursor agmatine, no increase in the levels of the polyamines putrescine, spermidine and spermine was detected in the transgenic plants. Similarly, the activities of ornithine decarboxylase and S-adenosylmethionine decarboxylase were unchanged. No diversion of polyamine metabolism into the hydroxycinnamic acid-polyamine conjugate pool or into the tobacco alkaloid nicotine was detected. Activity of the catabolic enzyme diamine oxidase was the same in transgenic and control plants. The elevated ADC activity and agmatine production were subjected to a metabolic/physical block preventing increased, i.e. deregulated, polyamine accumulation. Overaccumulation of agmatine in the transgenic plants did not affect morphological development. Publication Types: Research Support, Non-U.S. Gov't PMID: 9230111 [PubMed - indexed for MEDLINE] 2285: Hastings Cent Rep. 1997 Jul-Aug;27(4):34-8. Food biotechnology's challenge to cultural integrity and individual consent. Thompson PB. Texas A&M University, USA. Consumer response to genetically altered foods has been mixed in the United States. While transgenic crops have entered the food supply with little comment, other foods, such as the bioengineered tomato, have caused considerable controversy. Objections to genetically engineered food are varied, ranging from the religious to the aesthetic. One need not endorse these concerns to conclude that food biotechnology violates procedural protections of consumer sovereignty and religious liberty. Consumer sovereignty, a principle especially valued in this country, requires that information be made available so each individual or group may make food choices based on their own values. And as yet, there is no policy provision for informing consumers about the degree to which food has been genetically engineered. Publication Types: Review PMID: 9271720 [PubMed - indexed for MEDLINE] 2286: Transgenic Res. 1997 Jul;6(4):289-96. Expression of the human milk protein beta-casein in transgenic potato plants. Chong DK, Roberts W, Arakawa T, Illes K, Bagi G, Slattery CW, Langridge WH. Center for Molecular Biology and Gene Therapy, School of Medicine, Loma Linda, CA, USA. A 1177 bp cDNA fragment encoding the human milk protein beta-casein was introduced into Solanum tuberosum cells under control of the auxin-inducible, bidirectional mannopine synthase (mas1',2') promoters using Agrobacterium tumefaciens-mediated leaf disc transformation methods. Antibiotic-resistant plants were regenerated and transformants selected based on luciferase activity carried by the expression vector containing the human beta-casein cDNA. The presence of human beta-casein cDNA in the plant genome was detected by PCR and DNA hybridization experiments. Human beta-casein mRNA was identified in leaf tissues of transgenic plants by RT-PCR analysis. Human beta-casein was identified in auxin-induced leaf and tuber tissues of transformed potato plants by immunoprecipitation and immunoblot analysis. Human beta-casein produced in transgenic plants migrated in polyacrylamide gels as a single band with an approximate molecular mass of 30 kDa. Immunoblot experiments identified approximately 0.01% of the total soluble protein of transgenic potato leaf tissue as beta-casein. The above experiments demonstrate the expression of human milk beta-casein as part of an edible food plant. These findings open the way for reconstitution of human milk in edible plants for replacement of bovine milk in baby foods for general improvement of infant nutrition, and for prevention of gastric and intestinal diseases in children. Publication Types: Research Support, Non-U.S. Gov't PMID: 9232029 [PubMed - indexed for MEDLINE] 2287: Cas Lek Cesk. 1997 May 29;136(11):331-6. [The safety and usefulness of transgenic plants] [Article in Czech] Ondrej M, Drobník M. Ustav molekulární biologie rostlin AV CR, Ceské Budĕjovice. Transgenic crop plants, used in food and feed production, carry different beneficial transgenes, mostly for resistance to pests, herbicides and diseases. All new transgenic plant varieties, the genes they carry and their products have been thoroughly tested before released for agriculture and even more for marketing. Genetically modified organisms carry the same risk as any other organism. Food derived from genetically modified organisms due to legal regulation is most controlled and therefore most safe food ever placed on the market. In future, transgenic plants offer many new possibilities also for medical use, like plant vaccines, antibiotics and rare proteins of clinical importance produced by plants. Publication Types: English Abstract PMID: 9333501 [PubMed - indexed for MEDLINE] 2288: Plant J. 1997 May;11(5):1071-8. Transgenic rice (Oryza sativa) endosperm expressing daffodil (Narcissus pseudonarcissus) phytoene synthase accumulates phytoene, a key intermediate of provitamin A biosynthesis. Burkhardt PK, Beyer P, Wünn J, Klöti A, Armstrong GA, Schledz M, von Lintig J, Potrykus I. Swiss Federal Institute of Technology, Institute for Plant Sciences, Zürich, Switzerland. peter.burkhardt@ipw.biol.ethz.ch Rice (Oryza sativa L.), the major food staple for more than two billion people, contains neither beta-carotene (provitamin A) nor C40 carotenoid precursors thereof in its endosperm. To improve the nutritional value of rice, genetic engineering was chosen as a means to introduce the ability to make beta-carotene into rice endosperm tissue. Investigation of the biochemical properties of immature rice endosperm using [14C]-labelled substrates revealed the presence of geranyl geranyl diphosphate, the C20 general isoprenoid precursor necessary for C40 carotenoid biosynthesis. Phytoene synthase, which condenses two molecules of geranyl geranyl diphosphate, is the first of four specific enzymes necessary for beta-carotene biosynthesis in plants. Therefore, the Japonica rice model variety Taipei 309 was transformed by microprojectile bombardment with a cDNA coding for phytoene synthase from daffodil (Narcissus pseudonarcissus) under the control of either a constitutive or an endosperm-specific promoter. In transgenic rice plants, the daffodil enzyme is active, as measured by the in vivo accumulation of phytoene in rice endosperm. Thus, it is demonstrated for the first time that it is in principle possible to engineer a critical step in provitamin A biosynthesis in a non-photosynthetic, carotenoid-lacking plant tissue. These results have important implications for long-term prospects of overcoming worldwide vitamin A deficiency. Publication Types: Research Support, Non-U.S. Gov't PMID: 9193076 [PubMed - indexed for MEDLINE] 2289: J Nutr. 1997 May;127(5 Suppl):943S-947S. Genetic influences on the response of body fat and fat distribution to positive and negative energy balances in human identical twins. Bouchard C, Tremblay A. Physical Activity Sciences Laboratory, Laval University, Ste-Foy, Québec, Canada. This article summarizes a series of intervention studies conducted with pairs of young adult male identical twins and designed to determine whether there is any evidence for genotype x overfeeding or genotype x negative energy balance interaction effects in the changes in body weight, body composition, fat distribution, computerized tomography-assessed abdominal visceral fat, resting metabolic rate and thermic response to a standardized meal of mixed composition brought about by chronic exposure to appropriate experimental treatments. These studies demonstrated that individual differences in response to chronic alterations in energy balance are common. The comparison of the heterogeneity in response between the pairs of twins in contrast to the variance within pairs revealed that members of the same twin pair are significantly more alike than individuals who are not genetically related by descent. The intrapair resemblance in response was particularly strong for the changes in body mass, body composition, subcutaneous fat distribution and abdominal visceral fat. In contrast, the results of two long-term intervention studies showed that variations in resting metabolic rate following exposure to chronic overfeeding or negative energy balance induced by exercise were accounted for primarily by the changes in body mass. Finally, the thermic response to food was not modified by any of the experimental treatments. On the basis of these observations, we conclude that there are individuals at risk of gaining weight and body fat or who are resistant to weight loss. These differences in susceptibility to chronic overfeeding or in sensitivity to negative energy balance seem to be largely explained by genetic factors whose exact nature remains to be determined. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 9164270 [PubMed - indexed for MEDLINE] 2290: Int Arch Allergy Immunol. 1997 May-Jul;113(1-3):122-4. Recombinant proteins in newly developed foods: identification of allergenic activity. Lehrer SB, Reese G. Department of Medicine, Tulane University School of Medicine, New Orleans, La. 70012, USA. A number of agricultural crops are being modified for various purposes using recombinant DNA technology. Since transferred genes may code for proteins that are ordinarily not present, there is concern about the potential allergenicity of these new varieties. The safety evaluation of transgenic foods is relatively easy when the allergenicity of the gene source is known. Recombinant allergens in genetically engineered or altered foods can be identified using traditional immunological assays such as RAST or ELISA inhibition or immunoblotting procedures. Our recent studies of two corn proteins (10 kD and HSZ) used to alter grain amino acid composition and of transgenic soybeans with an altered fatty acid profile are examples of this approach. Both 10 kD and HSZ did not bind IgE antibodies from sera of corn-reactive subjects by immunoblotting. Studies of wild-type and transgenic soybeans with high oleic acidic content by RAST inhibition and immunoblotting with pooled sera of soy-allergic individuals demonstrated no difference in the allergen content of both extracts. In contrast to these studies, a recent investigation by Nordlee et al. (1996) of transgenic soybeans which expressed a methionine/cysteine-rich protein from Brazil nuts identified this protein as a major Brazil nut allergen. These studies indicate that, when the gene source is from a known allergen or if the recipient contains allergens, it is possible to determine whether the allergen content of the transgenic line is altered relative to the nontransgenic varieties. Publication Types: Research Support, Non-U.S. Gov't PMID: 9130499 [PubMed - indexed for MEDLINE] 2291: Rev Sci Tech. 1997 Apr;16(1):83-90. [The risk of transmission of salmonellae in poultry farming: detection and prevention in Europe] [Article in French] Humbert F, Salvat G. Centre national d'études vétérinaires et alimentaires, Unité de recherche et d'appui technique Hygiène et qualité des produits avicoles et porcins, Ploufragan, France. While salmonellas can cause disease problems among poultry, they remain essentially a concern for public health, as a cause of outbreaks of food poisoning. The principal site of multiplication of these bacteria is the digestive tract, particularly the caecum, which may result in widespread contamination of the environment. The pathogenicity of salmonellas depends on the invasive properties and the ability of the bacteria to survive and multiply within cells, particularly macrophages. These properties are the source of vertical transmission which, in the case of survival of the embryo, can result in contamination of a flock or, in the case of embryonic mortality, can result in an explosion of contaminated eggs. Salmonella infection can be diagnosed by isolating the bacteria and/or serological testing of the flock. European Union Directive 92/117/EC, modified by Directive 97/22/EC, stipulates either the destruction of infected flocks of breeding birds, or decontamination of the flock in an effective way, before normal trade in products can be resumed. Noteworthy examples of effective measures suitable for prophylaxis of Salmonella infection in poultry flocks include the slaughter of infected breeding stock, the creation of sanitary barriers at building entrances, heat treatment of feed, the use of competitive exclusion, selection of breeds genetically resistant to Salmonella, and occasional vaccination and antibiotic treatment. However, the most effective means of reducing food poisoning remains adequate cooking of food and maintenance of the cold chain. Publication Types: English Abstract Review PMID: 9537745 [PubMed - indexed for MEDLINE] 2292: Schweiz Med Wochenschr. 1997 Mar 29;127(13):554-60. [Is molecular biology useful to the practitioner?] [Article in French] Waeber G, Haefliger JA. Département de Médecine Interne B, CHUV, Lausanne. The relative importance of molecular biology in clinical practice is often underestimated. However, numerous procedures in clinical diagnosis and new therapeutic drugs have resulted from basic molecular research. Furthermore, understanding of the physiological and physiopathological mechanisms underlying several human diseases has been improved by the results of basic molecular research. For example, cloning of the gene encoding leptin has provided spectacular insights into the understanding of the mechanisms involved in the control of food intake and body weight maintenance in man. In cystic fibrosis, the cloning and identification of several mutations in the gene encoding the chloride channel transmembrane regulator (CFTR) have resolved several important issues in clinical practice: cystic fibrosis constitutes a molecular defect of a single gene. There is a strong correlation between the clinical manifestations or the severity of the disease (phenotype) with the type of mutations present in the CFTR gene (genotype). More recently, identification of mutations in the gene encoding a subunit of the renal sodium channel in the Liddle syndrome has provided important insight into the physiopathological understanding of mechanisms involved in this form of hereditary hypertension. Salt retention and secondary high blood pressure are the result of constitutive activation of the renal sodium channel by mutations in the gene encoding the renal sodium channel. It is speculated that less severe mutations in this channel could result in a less severe form of hypertension which may correspond to patients suffering from high blood pressure with low plasma renin activity. Several tools, most notably PCR, are derived from molecular research and are used in everyday practice, i.e. in prenatal diagnosis and in the diagnosis of several infectious diseases including tuberculosis and hepatitis. Finally, the production of recombinant proteins at lower cost and with fewer side effects is used in everyday clinical practice. Gene therapy remains an extraordinary challenge in correcting severe hereditary or acquired diseases. The use of genetically modified animal cell lines producing growth factors, insulin or erythropoetin, which are subsequently encapsulated and transferred to man, represents an attractive approach for gene therapy. Publication Types: English Abstract Review PMID: 9190668 [PubMed - indexed for MEDLINE] 2293: Lakartidningen. 1997 Feb 19;94(8):612-7. [Does genetically altered food cause allergy and poisoning? High safety requirements could eliminate health hazards] [Article in Swedish] Wilhelmson B. PMID: 9072649 [PubMed - indexed for MEDLINE] 2294: Fed Regist. 1997 Feb 14;62(31):7108-23. Recombinant DNA research: proposed actions under the guidelines; notice. U.S. National Institutes of Health. Publication Types: Government Publications PMID: 11654870 [PubMed - indexed for MEDLINE] 2295: Int J Biol Macromol. 1997 Feb;20(1):1-7. Effect of o-acyl substituents on the functional behaviour of Rhizobium meliloti succinoglycan. Ridout MJ, Brownsey GJ, York GM, Walker GC, Morris VJ. Institute of Food Research, Norwich Laboratory, Colney, UK. The effects of selective removal of acetyl or succinyl substituents on the functionality of succinoglycan polysaccharide have been studied by comparing the behaviour of the polysaccharides isolated from native Rhizobium meliloti strain Rm1021, and genetically modified R. meliloti species. Removal of the succinyl groups was found to dramatically improve pseudoplasticity of the aqueous succinoglycan samples and also increase the cooperativity of the order-disorder transition exhibited by the polysaccharide. Removal of the acetyl substituent led to a decrease in the order-disorder transition temperature, whereas the removal of the succinyl groups led to an increase. Publication Types: Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. PMID: 9110180 [PubMed - indexed for MEDLINE] 2296: Nat Biotechnol. 1997 Feb;15(2):111. The end of big government? Miller HI. Hoover Institution, USA. miller@hoover.stanford.edu Publication Types: Research Support, Non-U.S. Gov't PMID: 9035118 [PubMed - indexed for MEDLINE] 2297: Wien Klin Wochenschr. 1997 Jan 31;109(2):33-9. [Transgenic foods--advantages and risks] [Article in German] Mannhalter C. Klinisches Institut für Medizinische und Chemische Labordiagnostik, Bereich Molekularbiologie, Medizinische Fakultät, Universität Wien, Osterreich. Publication Types: Review PMID: 9123942 [PubMed - indexed for MEDLINE] 2298: Nature. 1997 Jan 23;385(6614):290. Comment in: Nature. 1997 Mar 13;386(6621):111. Comment on: Nature. 1996 Dec 12;384(6609):502-3. Pros and cons of foreign genes in crops. Ort DR. Publication Types: Comment Letter PMID: 9002507 [PubMed - indexed for MEDLINE] 2299: Tidsskr Nor Laegeforen. 1997 Jan 10;117(1):84. "Contamination" with genes?] [Article in Norwegian] Naess K. PMID: 9064819 [PubMed - indexed for MEDLINE] 2300: Sci Technol Human Values. 1997 Winter;22(1):98-124. Public concerns in the United Kingdom about general and specific applications of genetic engineering: risk, benefit, and ethics. Frewer LJ, Howard C, Shepherd R. PMID: 11654686 [PubMed - indexed for MEDLINE] 2301: Food Drug Law J. 1997;52(1):49-60. The food label and the right-to-know. Degnan FH. King & Spalding, Washington, DC, USA. PMID: 10346709 [PubMed - indexed for MEDLINE] 2302: Bioseparation. 1997;7(1):31-7. The application of aqueous two-phase systems to the purification of pharmaceutical proteins from transgenic sheep milk. Harris DP, Andrews AT, Wright G, Pyle DL, Asenjo JA. Dept. Food Science & Technology University of Reading, UK. Transgenic sheep milk containing the protein human alpha 1-Antitrypsin (AAT) was partitioned in Poly(ethylene glycol) (PEG)-Sulphate and PEG-Phosphate biphasic systems. Individual partition coefficients for AAT and some of the milk proteins were determined in these systems. The effects of PEG molecular weight, pH and the inclusion of NaCl on the partitioning of the proteins were also studied. It was found that increasing the concentration of NaCl and decreasing the molecular weight of the PEG resulted in an increase of the partition coefficients of the proteins to the upper (PEG) phase. This partitioning effect was greater for the more hydrophobic proteins and particularly in systems having a pH close to the isoelectric point of the protein. Solubilities of the proteins in increasing concentrations of ammonium sulphate were measured in order to investigate the effects of hydrophobic and electrostatic interactions on the partitioning of these proteins in aqueous two-phase systems. Those proteins that precipitated at low levels of ammonium sulphate showed an increase in partition coefficient at low concentrations of NaCl, or they were precipitated at the interface of the phase at low concentrations of NaCl. Proteins that had low salting out constants in ammonium sulphate solutions were relatively unaffected by NaCl in ATPS. It is probable however that conformational changes and the state of aggregation of proteins are also important and should be invoked in describing the partitioning behavior observed for beta-Lg for example. Comparison of theoretical and experimental values for AAT yield and purity showed clearly that partition coefficients are influenced by the degree of purity and values obtained with purified standards are not necessarily the same as for the same protein present in a complex mixture. Under the most favourable conditions using a 4% w/w loading of transgenic ovine milk, we obtained a 91% yield of AAT in the PEG phase with a purity of 73%. PMID: 9615611 [PubMed - indexed for MEDLINE] 2303: Peptides. 1997;18(6):825-33. Further evidence that the tachykinin PG-KII is a potent agonist at central NK-3, but not NK-1, receptors. Polidori C, Panocka I, Ciccocioppo R, Broccardo M, Improta G, Regoli D, Massi M. Department of Pharmacological sciences and Experimental Medicine, University of Camerino, Italy. polidori@cambio.unicam.it Intracerebroventricular (i.c.v.) injection of tachykinins (TKs) inhibits ethanol intake and angiotensin II-induced water intake; the effects are apparently mediated by NK-3 and NK-1 receptors, respectively. The present study evaluated the effect of the TK PG-KII, a novel kassinin-like peptide isolated from the skin of the Australian frog Pseudophryne güntheri, in these in vivo tests for central activity. PG-KII, given by i.c.v. injection, potently inhibited alcohol intake in genetically selected alcohol-preferring rats, being about 3 times more potent than the selective NK-3 receptor agonist NH2-SENK. The dose of 100 ng/rat, that markedly inhibited ethanol intake, did not inhibit food intake and prandial drinking in food deprived rats, providing evidence that the effect of PG-KII on ethanol intake is behaviorally selective. The effect on ethanol intake was inhibited by i.c.v. injection of the NK-3 receptor antagonist R820, but was not modified by the NK-1 receptor antagonist SR 140333. PG-KII inhibited drinking induced by angiotensin II only at doses of 300 or 1000 ng/rat, being about 5 times less potent than the selective NK-1 receptor agonist [Sar9, Met(O2)11] substance P. These doses of PG-KII produced also marked increase in competing behaviors, such as grooming and locomotion. The dose of 1000 ng/rat evoked a general inhibition of the ingestive behavior, reducing also food intake. The i.c.v. injection of the NK-1 receptor antagonist SR 140,333 only slightly inhibited the effect of PG-KII on angiotensin II-induced drinking, while it markedly reduced that of [Sar9, Met(O2)11] substance P. These findings, in accordance with those of previous studies, indicate that PG-KII is endowed with marked activity at central NK-3 receptors, and low activity at NK-1 receptors. Publication Types: Research Support, Non-U.S. Gov't PMID: 9285931 [PubMed - indexed for MEDLINE] 2304: Nat Biotechnol. 1997 Jan;15(1):2. Commissioner Kessler's agbio legacy. Hoyle R. PMID: 9035091 [PubMed - indexed for MEDLINE] 2305: Econ Polit Wkly. 1996 Dec 14;31(50):3231-2. Bogus debate on bioethics. Sahai S. PMID: 11660559 [PubMed - indexed for MEDLINE] 2306: Nat Biotechnol. 1996 Dec;14(13):1628. Taking the hex off transgenic plant exports. Hoyle R. PMID: 9634830 [PubMed - indexed for MEDLINE] 2307: Endocrinol Metab Clin North Am. 1996 Dec;25(4):781-800. Lessons from animal models of obesity. York DA. Experimental Obesity Research Program, Pennington Biomedical Research Center, Louisiana State University, Baton Rouge, USA. Obesity in animals may result from genetic, dietary, or neuroendocrine perturbations. Study of these models has identified the central systems that regulate food intake and energy expenditure and identified the interdependence of feeding behavior, the autonomic nervous system, and adrenal glucocorticoids in the development of obesity. The animal models of obesity have been influential in showing that adipose tissue is an important secretory tissue. The recent identification of five genes that cause obesity will provide new insight into the physiologic systems that regulate energy balance. Publication Types: Research Support, U.S. Gov't, P.H.S. Review PMID: 8977045 [PubMed - indexed for MEDLINE] 2308: Science. 1996 Nov 22;274(5291):1389-91. Control of C. elegans larval development by neuronal expression of a TGF-beta homolog. Ren P, Lim CS, Johnsen R, Albert PS, Pilgrim D, Riddle DL. Molecular Biology Program and Division of Biological Sciences, 311 Tucker Hall, University of Missouri, Columbia, MO 65211, USA. riddle@biosci.mbp.missouri.edu The Caenorhabditis elegans dauer larva is specialized for dispersal without growth and is formed under conditions of overcrowding and limited food. The daf-7 gene, required for transducing environmental cues that support continuous development with plentiful food, encodes a transforming growth factor-beta (TGF-beta) superfamily member. A daf-7 reporter construct is expressed in the ASI chemosensory neurons. Dauer-inducing pheromone inhibits daf-7 expression and promotes dauer formation, whereas food reactivates daf-7 expression and promotes recovery from the dauer state. When the food/pheromone ratio is high, the level of daf-7 mRNA peaks during the L1 larval stage, when commitment to non-dauer development is made. Publication Types: Research Support, U.S. Gov't, P.H.S. PMID: 8910282 [PubMed - indexed for MEDLINE] 2309: Mutat Res. 1996 Nov 11;372(1):53-64. Molecular analysis of lacI mutations in Rat2 cells exposed to 7,12-dimethylbenz[a]anthracene: evidence for DNA sequence and DNA strand biases for mutation. Manjanatha MG, Chen JB, Shaddock JG Jr, Harris AJ, Shelton SD, Casciano DA. Department of Health and Human Services, Food and Drug Administration, National Center for Toxicological Research, Jefferson, AR 72079, USA. mmanjanatha@nctr.fda.gov The Rat2 cell line carries 50-70 stably integrated copies per cell of a lambda/lacI shuttle vector as a target for mutagenicity testing. Rat2 cells were exposed to 1 and 10 micrograms/ml of 7,12-dimethylbenz[a]anthracene (DMBA) for 24 h at 37 degrees C in the presence of primary rat hepatocytes, and grown to confluence. The shuttle vector was rescued from untreated and mutagen-treated cells and mutant frequencies were determined. The low and high doses of DMBA induced mutant frequencies that were 7-fold (25 +/- 4.9 x 10(-5)) and 33-fold (127 +/- 19.9 x 10(-5)) higher, respectively, than the spontaneous mutant frequency (3.8 +/- 0.7 x 10(-5)). DNA sequence analysis of the DMBA-induced lacI- mutants indicated that they contained mainly basepair substitution mutations at A:T and G:C, and that A:T-->T:A and G:C-->T:A transversions were the predominant types. In addition, 23 of 28 (82%) A:T basepair substitution mutations occurred with the mutated dA, the putatively adducted base, on the coding strand. Furthermore, 20 of the 28 (71%) A:T mutations had the mutated dA flanked 5' by a dC, and 17 of these were A:T-->T:A transversions, suggesting a sequence preference for this mutation. Except for a higher proportion of G:C-->A:T transitions in the low dose data, the mutational profiles from low and high doses of DMBA were similar. These results indicate that DMBA mutagenesis in the lacI gene of Rat2 cells displays distinct DNA sequence and DNA strand preferences. Publication Types: Comparative Study PMID: 9003531 [PubMed - indexed for MEDLINE] 2310: Med Law Rev. 1996 Autumn;4(3):250-69. The pig, the transplant surgeon and the Nuffield Council. Cartwright W. PMID: 11657151 [PubMed - indexed for MEDLINE] 2311: Nat Biotechnol. 1996 Oct;14(10):1269-73. Stability of food allergens to digestion in vitro. Astwood JD, Leach JN, Fuchs RL. Monsanto Company, St. Louis, MO 63198, USA. An integral part of the safety assessment of genetically modified plants is consideration of possible human health effects, especially food allergy. Prospective testing for allergenicity of proteins obtained from sources with no prior history of causing allergy has been difficult because of the absence of valid methods and models. Food allergens may share physicochemical properties that distinguish them from nonallergens, properties that may be used as a tool to predict the inherent allergenicity of proteins newly introduced into the food supply by genetic engineering. One candidate property is stability to digestion. We have systematically evaluated the stability of food allergens that are active via the gastrointestinal tract in a simple model of gastric digestion, emphasizing the major allergens of plant-derived foods such as legumes (peanuts and soybean). Important food allergens were stable to digestion in the gastric model (simulated gastric fluid). For example, soybean beta-conglycinin was stable for 60 min. In contrast, nonallergenic food proteins, such as spinach ribulose bis-phosphate carboxylase/oxygenase, were digested in simulated gastric fluid within 15 sec. The data are consistent with the hypothesis that food allergens must exhibit sufficient gastric stability to reach the intestinal mucosa where absorption and sensitization (development of atopy) can occur. Thus, the stability to digestion is a significant and valid parameter that distinguishes food allergens from nonallergens. Publication Types: In Vitro PMID: 9631091 [PubMed - indexed for MEDLINE] 2312: Allerg Immunol (Paris). 1996 Oct;28(8):288-9. [Questions about transgenic foods--importance and/or dangers-- importance of research on unexpected cross-reactions] [Article in French] Sabbah A. Laboratoire d'Immuno-Allergologie, CHU, Angers. Publication Types: Review PMID: 9011168 [PubMed - indexed for MEDLINE] 2313: Antonie Van Leeuwenhoek. 1996 Oct;70(2-4):299-316. Barriers to application of genetically modified lactic acid bacteria. Verrips CT, van den Berg DJ. Unilever Research Laboratorium Vlaardingen, Vlaardingen The Netherlands. To increase the acceptability of food products containing genetically modified microorganisms it is necessary to provide in an early stage to the consumers that the product is safe and that the product provide a clear benefit to the consumer. To comply with the first requirement a systematic approach to analyze the probability that genetically modified lactic acid bacteria will transform other inhabitants of the gastro- intestinal (G/I) tract or that these lactic acid bacteria will pick up genetic information of these inhabitants has been proposed and worked out to some degree. From this analysis it is clear that reliable data are still missing to carry out complete risk assessment. However, on the basis of present knowledge, lactic acid bacteria containing conjugative plasmids should be avoided. Various studies show that consumers in developed countries will accept these products when they offer to them health or taste benefits or a better keepability. For the developing countries the biggest challenge for scientists is most likely to make indigenous fermented food products with strongly improved microbiological stability due to broad spectra bacteriocins produced by lactic acid bacteria. Moreover, these lactic acid bacteria may contribute to health. Publication Types: Review PMID: 8879412 [PubMed - indexed for MEDLINE] 2314: Nat Biotechnol. 1996 Sep;14(9):1155-9. Integration and expression of the high-molecular-weight glutenin subunit 1Ax1 gene into wheat. Altpeter F, Vasil V, Srivastava V, Vasil IK. Laboratory of Plant Cell and Molecular Biology, University of Florida, Gainesville 32611-0690, USA. The unique bread-making characteristic of wheat flour is closely related to the elasticity and extensibility of the gluten proteins stored in the starchy endosperm, particularly the high-molecular-weight glutenin subunits (HMW-GS), which are important in determining gluten and dough elasticity. The quality of wheat cultivars depends on the number and composition of the HMW-GS present. We have introduced the HMW-GS 1Ax1 gene, known to be associated with good bread-making quality, into the Bob White cultivar of wheat (Triticum aestivum L.), in which it is not present in nature, by the biolistic bombardment of cultured immature embryos. Of the 21 independent transformed lines selected, 20 expressed the selectable bar gene, and nine the 1Ax1 gene. The amount of HMW-GS 1Ax1 protein produced in the different transgenic lines varied from 0.6% to 2.3% of the total protein, resulting in an increase of up to 71% in total HMW-GS proteins. The transgenic plants were normal, fertile, and showed Mendelian segregation of the transgenes. The accumulation of HMW-GS 1Ax1 was consistent and stable up to the R3 seed generation. These results demonstrate that it is possible to manipulate both the quantity and quality of HMW-GS, which influence the bread-making quality of wheat. Publication Types: Research Support, Non-U.S. Gov't PMID: 9631070 [PubMed - indexed for MEDLINE] 2315: Eur J Biochem. 1996 Jul 15;239(2):281-93. Bean ribonuclease-like pathogenesis-related protein genes (Ypr10) display complex patterns of developmental, dark-induced and exogenous-stimulus-dependent expression. Walter MH, Liu JW, Wünn J, Hess D. Universität Hohenheim, Institut für Pflanzenphysiologie, Stuttgart, Germany. The intracellular pathogenesis-related (PR) proteins of common bean (Phaseolus vulgaris L.) are encoded by a highly polymorphic family of at least 20 genes. One member, the Ypr10*c gene, has been isolated and characterised. The deduced amino acid sequence of the encoded protein, PR-10, exhibits similarities to tree-pollen allergens, to food allergens from celery and apple and to ginseng ribonuclease peptide sequences. We show by RNA blot analysis that the Ypr10 gene family, including Ypr10*c, is strongly expressed in bean roots. In leaves Ypr10 transcript levels are low in young and mature stages but are elevated during senescence and in diseased states. Dark treatment of leaves causes strong induction of Ypr10 transcripts, which is reversible by light, and diurnal rhythms of transcript accumulation during the night are observed. Ypr10 genes are responsive to external stimuli related to pathogen-defence such as glutathione or salicylic acid. Transcriptional activity of a Ypr10*c promoter-beta-glucuronidase fusion gene in transgenic tobacco was observed in roots, in developing xylem and phloem of stems, and in the blade of senescent leaves, with highest levels at the onset of senescence. The most striking characteristic of developmental expression was the specific localisation of beta-glucuronidase activity in the transmitting tract of styles in flowers at anthesis. Feeding of various pathogen-related and stress-related stimuli to young tobacco leaves led to accumulation of GUS activity in leaf blades. We identify considerable spatio-temporal similarities between reported expression patterns of Ypr10 genes and ribonuclease genes, which, together with the significant sequence similarity to the ginseng ribonuclease, support the hypothesis of a ribonuclease function for PR-10 proteins and allow the prediction of possible biological roles. Publication Types: Comparative Study Research Support, Non-U.S. Gov't PMID: 8706731 [PubMed - indexed for MEDLINE] 2316: Genethics News. 1996 Jul-Aug;13:6-7. Genethics in the mid-1990s. [No authors listed] Publication Types: News PMID: 11660393 [PubMed - indexed for MEDLINE] 2317: Risk. 1996 Summer;7(3):253-66. Impact of the Human Genome Project at the interface between patent and FDA laws. Cunningham BC. PMID: 11655078 [PubMed - indexed for MEDLINE] 2318: Int J Food Microbiol. 1996 Jul;30(3):315-24. Genetically modified starter and protective cultures. Geisen R, Holzapfel WH. Federal Research Centre for Nutrition, Institute of Hygiene and Toxicology, Karlsruhe, Germany. Modern approaches towards starter and protective culture improvement rely on advances in molecular biology. For most microorganisms used for food production, gene technological methods have been well developed. By recombinant DNA technology, 'tailor-made' starter and protective cultures may be constructed so as to combine technically desirable features. A single strain which normally would fail to accomplish a given 'task' may now be improved so as to meet a set of requirements necessary for a specific production or preservation process (e.g. wholesomeness, no off-flavour production, overproduction of bacteriocins or particular enzymes). In addition, undesirable properties (e.g. mycotoxin or antibiotic production by cheese moulds) may be eliminated by techniques such as 'gene disruption'. PMID: 8854184 [PubMed - indexed for MEDLINE] 2319: Nat Biotechnol. 1996 Jun;14(6):736-40. Genetic transformation of cassava (Manihot esculenta Crantz). Li HQ, Sautter C, Potrykus I, Puonti-Kaerlas J. Inst. of Plant Sciences, Swiss Federal Institute of Technology, ETH Zentrum, Zürich, Switzerland. Genetic engineering can be used to complement traditional breeding methods in crop plant improvement. Transfer of genes from heterologous species provides the means of selectively introducing new traits into crop plants and expanding the gene pool beyond what has been available to traditional breeding systems. The prerequisites for genetic engineering are efficient transformation and tissue culture systems that allow selection and regeneration of transgenic plants. Cassava, an integral plant for food security in developing countries, has until now been recalcitrant to transformation approaches. We report here a method for regenerating stably transformed cassava plants after cocultivation with Agrobacterium tumefaciens, which opens cassava for future improvement via biotechnology. Publication Types: Research Support, Non-U.S. Gov't PMID: 9630981 [PubMed - indexed for MEDLINE] 2320: Proc Biol Sci. 1996 May 22;263(1370):647-51. Growth hormone increases predation exposure of rainbow trout. Jönsson E, Johnsson JI, Björnsson BT. Department of Zoophysiology, Göteborg University, Sweden. The energetic state of an animal strongly influences decisions that balances feeding against predation risk. Growth hormone increases the metabolic demands, which should elevate the feeding motivation of an animal. This, in turn, may increase the willingness to risk exposure to predators during feeding. To test this hypothesis, we studied the effect of growth hormone on the behavioural response of rainbow trout (Oncorhynchus mykiss) to simulated attacks from a model heron. After attacks, growth hormone treated trout foraged closer to the water surface, resumed feeding earlier, and ate more food than did control trout. Such behaviour should increase the susceptibility to aerial predation. Thus, predation may select against high endogenous growth hormone secretion in wild fish. Furthermore, genetic manipulations to increase growth hormone levels, intended to improve growth performance in aquaculture, may result in individuals with substantially altered behavioural patterns. In light of the increasing potential for interactions between farmed and wild fish, growth hormone transgenic fish may pose a threat to wild fish populations. Publication Types: Research Support, Non-U.S. Gov't PMID: 8677262 [PubMed - indexed for MEDLINE] 2321: Artif Organs. 1996 May;20(5):396-402. Ex vivo manipulation of cell subsets for cell therapies. Nordon RE, Schindhelm K. Graduate School of Biomedical Engineering, University of New South Wales, Sydney, Australia. Large-scale cell separation and ex vivo expansion technologies will form the basis for development of new cellular products for the treatment of cancer and fatal viral diseases. The cell subsets that are likely to play a significant role in cellular therapy include hematopoietic stem cells, platelet and granulocyte precursors, cytotoxic lymphocytes, and genetically modified hematopoietic or lymphoid precursors. Cell enrichment techniques are required to eliminate tumor cells from autologous stem cell grafts and to reduce the size of culture systems required for expansion or gene transfection. The consumption of expensive culture components such as cytokines and serum may be reduced by the use of perfusion bioreactor devices. Methods that have been developed for the production of cell subsets for cellular therapy are reviewed. Publication Types: Review PMID: 8725617 [PubMed - indexed for MEDLINE] 2322: Transfus Med Rev. 1996 Apr;10(2):131-43. Blood proteins from transgenic animal bioreactors. Luboń H, Paleyanda RK, Velander WH, Drohan WN. Jerome H. Holland Laboratory, Plasma Derivatives Department, American Red Cross, Rockville, MD, USA. Publication Types: Review PMID: 8721970 [PubMed - indexed for MEDLINE] 2323: Am J Clin Nutr. 1996 Apr;63(4):651S-6S. Genetic modification of plants: significant issues and hurdles to success. Day PR. Center for Agricultural Molecular Biology, Rutgers, The State University of New Jersey, New Brunswick, 08902, U.S.A. Transformation and regeneration is routine for many crop plants. A genetically engineered tomato with a longer shelf life at full ripeness was introduced in the United States in 1994, and other soon-to-be-released products, both foods and fibers, incorporate genes for resistance to pests, diseases, and environmentally benign herbicides. Other possibilities are altered plant fats and oils, methionine- and lysine-enhanced grain and legume proteins, plant foods that can deliver immunizing antigens, and other ways of controlling fruit ripening. Food safety concerns include the inadvertent production of toxicants and allergens. Foreign DNA can be introduced into plants by bacterial vectors, direct uptake by protoplasts, and mechanical introduction on metal particles or other materials. Limitations include little or no control of copy number or site of integration of the introduced DNA, dependence on selectable markers for recovery of traits, and inadequate knowledge of how to control key metabolic steps to maximize desirable traits. Directed genetic change still requires conventional crop breeding to deliver benefits to farmers and consumers. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 8599334 [PubMed - indexed for MEDLINE] 2324: Am J Clin Nutr. 1996 Apr;63(4):622S-6S. Recombinant human milk proteins - an opportunity and a challenge. Lönnerdal B. Departnemt of Nutrition, University of California at Davis, 95616, U.S.A. Several human milk proteins have physiologic functions in infants. These proteins are involved in defense against infectious agents and in the optimization of nutrient uptake from milk. Therefore, interest in producing recombinant human milk proteins to use in infant formula has been growing. Microorganisms and transgenic animals can now be used for the production of bioactive proteins. However, the benefits of each protein must be evaluated in cells, animal models, and infants before claims can be made that adding them to formula improves the health or nutrition of infants. Once benefits are shown, proper manufacturing conditions must be developed for introducing the protein or proteins into formula. Processing conditions must be evaluated to ensure that biologic activity is maintained. Dry blending, aseptic processing, sterile filtration, and other techniques will likely be necessary for introducing proteins that require specific tertiary structure for activity. The importance of posttranslational modifications must also be considered: some proteins may require proper glycosylation or phosphorylation for physiologic activity. Publication Types: Review PMID: 8599329 [PubMed - indexed for MEDLINE] 2325: N Engl J Med. 1996 Mar 14;334(11):726-8. Comment on: N Engl J Med. 1996 Mar 14;334(11):688-92. Allergies to transgenic foods--questions of policy. Nestle M. Publication Types: Comment Editorial PMID: 8594435 [PubMed - indexed for MEDLINE] 2326: N Engl J Med. 1996 Mar 14;334(11):688-92. Comment in: N Engl J Med. 1996 Mar 14;334(11):726-8. Identification of a Brazil-nut allergen in transgenic soybeans. Nordlee JA, Taylor SL, Townsend JA, Thomas LA, Bush RK. Department of Food Science and Technology, University of Nebraska, Lincoln, 68583-0919, USA. BACKGROUND. The nutritional quality of soybeans (Glycine max) is compromised by a relative deficiency of methionine in the protein fraction of the seeds. To improve the nutritional quality, methionine-rich 2S albumin from the Brazil nut (Betholletia excelsa) has been introduced into transgenic soybeans. Since the Brazil nut is a known allergenic food, we assessed the allergenicity of the 2S albumin. METHODS. The ability of proteins in transgenic and non-transgenic soybeans, Brazil nuts, and purified 2S albumin to bind to IgE in serum from subjects allergic to Brazil nuts was determined by radioallergosorbent tests (4 subjects) and sodium dodecyl sulfate-polyacrylamide-gel electrophoresis (9 subjects) with immunoblotting and autoradiography. Three subjects also underwent skin-prick testing with extracts of soybean, transgenic soybean, and Brazil nut. RESULTS. On radioallergosorbent testing of pooled serum from four subjects allergic to Brazil nuts, protein extracts of transgenic soybean inhibited binding of IgE to Brazil-nut proteins. On immunoblotting, serum IgE from eight of nine subjects bound to purified 2S albumin from the Brazil nut and the transgenic soybean. On skin-prick testing, three subjects had positive reactions to extracts of Brazil nut and transgenic soybean and negative reactions to soybean extract. CONCLUSIONS. The 2S albumin is probably a major Brazil-nut allergen, and the transgenic soybeans analyzed in this study contain this protein. Our study show that an allergen from a food known to be allergenic can be transferred into another food by genetic engineering. Publication Types: Research Support, Non-U.S. Gov't PMID: 8594427 [PubMed - indexed for MEDLINE] 2327: Politics Life Sci. 1996 Mar;15(1):117-9. Ethical aspects of the labelling of foods derived from modern biotechnology. Lenoir N; European Commission. Group of Advisers on the Ethical Implications of Biotechnology, et al. PMID: 11655011 [PubMed - indexed for MEDLINE] 2328: Transgenic Res. 1996 Mar;5(2):105-113. A system for tissue-specific copper-controllable gene expression in transgenic plants: nodule-specific antisense of aspartate aminotransferase-P2. Mett VL, Podivinsky E, Tennant AM, Lochhead LP, Jones WT, Reynolds PH. Plant Improvement Division, Horticulture and Food Research Institute of New Zealand, Palmerston North. A vector system, based on copper controllable gene expression, has been developed to give control over place as well as time of expression of an introduced gene. This system consists of two elements: (1) the yeast ace1 gene encoding a metallo-regulatory transcription factor, ACE1, under control of either an organ-specific or a constitutive promoter; and (2) a gene of interest under control of a chimaeric promoter consisting of the 46 bp TATA fragment of the CaMV 35S RNA promoter linked to four repeats of the ACE1 binding site. The functioning of the system in an organ-specific manner was tested in nodulated Lotus corniculatus plants which consisted of non-transformed shoots plus transformed hairy root tissue 'wild-type tops/transgenic roots'. After addition of copper ions to the plant nutrient solution, beta-glucuronidase (GUS) expression was visualized either specifically in nodules or in both roots and nodules when the ace1 gene was placed under control of the nod45 promoter or the CaMV 35S RNA promoter, respectively. The nodule-specific system was used to express antisense constructs of aspartate aminotransferase-P2 in transgenic Lotus corniculatus plants. When expression was induced by the addition of copper ions to the plant nutrient solution aspartate aminotransferase-P2 activity declined dramatically, and a decrease of up to 90% was observed in nodule asparagine concentration. PMID: 8866892 [PubMed - indexed for MEDLINE] 2329: J Nutr. 1996 Mar;126(3):717-27. The feeding value of soybeans fed to rats, chickens, catfish and dairy cattle is not altered by genetic incorporation of glyphosate tolerance. Hammond BG, Vicini JL, Hartnell GF, Naylor MW, Knight CD, Robinson EH, Fuchs RL, Padgette SR. Monsanto Company, St. Louis, MO 63167, USA. Animal feeding studies were conducted with rats, broiler chickens, catfish and dairy cows as part of a safety assessment program for a soybean variety genetically modified to tolerate in-season application of glyphosate. These studies were designed to compare the feeding value (wholesomeness) of two lines of glyphosate-tolerant soybeans (GTS) to the feeding value of the parental cultivar from which they were derived. Processed GTS meal was incorporated into the diets at the same concentrations as used commercially; diary cows were fed 10 g/100 g cracked soybeans in the diet, a level that is on the high end of what is normally fed commercially. In a separate study, laboratory rats were fed 5 and 10 g unprocessed soybean meal 100 g diet. The study durations were 4 wk (rats and dairy cows), 6 wk (broilers) and 10 wk (catfish). Growth, feed conversion (rats, catfish, broilers), fillet composition (catfish), and breast muscle and fat pad weights (broilers) were compared for animals fed the parental and GTS lines. Milk production, milk composition, rumen fermentation and nitrogen digestibility were also compared for dairy cows. In all studies, measured variables were similar for animals fed both GTS lines and the parental line, indicating that the feeding value of the two GTS lines is comparable to that of the parental line. These studies support detailed compositional analysis of the GTS seeds, which showed no meaningful differences between the parental and GTS lines in the concentrations of important nutrients and antinutrients. They also confirmed the results of other studies that demonstrated the safety of the introduced protein, a bacterial 5-enolpyruvyl-shikimate-3-phosphate synthase from Agrobacterium sp. strain CP4. PMID: 8598557 [PubMed - indexed for MEDLINE] 2330: J Nutr. 1996 Mar;126(3):702-16. The composition of glyphosate-tolerant soybean seeds is equivalent to that of conventional soybeans. Padgette SR, Taylor NB, Nida DL, Bailey MR, MacDonald J, Holden LR, Fuchs RL. Monsanto Company, St. Louis, MO 63198, USA. One important aspect of the safety assessment of genetically engineered crops destined for food and feed uses is the characterization of the consumed portion of the crop. One crop currently under development, glyphosate-tolerant soybeans (GTS), was modified by the addition of a glyphosate-tolerance gene to a commercial soybean cultivar. The composition of seeds and selected processing fractions from two GTS lines, designated 40-3-2 and 61-67-1, was compared with that of the parental soybean cultivar, A5403. Nutrients measured in the soybean seeds included macronutrients by proximate analyses (protein, fat, fiber, ash, carbohydrates), amino acids and fatty acids. Antinutrients measured in either the seed or toasted meal were trypsin inhibitor, lectins, isoflavones, stachyose, raffinose and phytate. Proximate analyses were also performed on batches of defatted toasted meal, defatted nontoasted meal, protein isolate, and protein concentrate prepared from GTS and control soybean seeds. In addition, refined, bleached, deodorized oil was made, along with crude soybean lecithin, from GTS and control soybeans. The analytical results demonstrated the GTS lines are equivalent to the parental, conventional soybean cultivar. PMID: 8598556 [PubMed - indexed for MEDLINE] 2331: Crit Rev Food Sci Nutr. 1996;36 Suppl:S165-86. Assessment of the allergenic potential of foods derived from genetically engineered crop plants. Metcalfe DD, Astwood JD, Townsend R, Sampson HA, Taylor SL, Fuchs RL. ILSI Allergy and Immunology Institute, USA. This article provides a science-based, decision tree approach to assess the allergenic concerns associated with the introduction of gene products into new plant varieties. The assessment focuses on the source from which the transferred gene was derived. Sources fall into three general categories: common allergenic food proteins; less common allergenic foods or other known allergen sources; and sources with no history of allergenicity. Information concerning the amino acid sequence identity to known allergenic proteins, in vitro and/or in vivo immunologic assays, and assessment of key physiochemical properties are included in reaching a recommendation on whether food derived from the genetically modified plant variety should be labeled as to the source of the transferred gene. In the end, a balanced judgement of all the available data generated during allergenicity assessment will assure the safety of foods derived from genetically engineered crops. Using the approaches described here, new plant varieties generated by genetic modification should be introduced into the marketplace with the same confidence that new plant varieties developed by traditional breeding have been introduced for decades. Publication Types: Review PMID: 8959382 [PubMed - indexed for MEDLINE] 2332: Monogr Allergy. 1996;32:105-20. Allergenicity of foods derived from transgenic plants. Astwood JD, Fuchs RL. Ceregen, Monsanto Company, St. Louis, MO, USA. PMID: 8813190 [PubMed - indexed for MEDLINE] 2333: Planta. 1996;198(2):246-52. Soluble acid invertase determines the hexose-to-sucrose ratio in cold-stored potato tubers. Zrenner R, Schüler K, Sonnewald U. Institut für Pflanzengenetik und Kulturpflanzenforschung, Gatersleben, Germany. Cold storage of potato (Solanum tuberosum L.) tubers is known to cause accumulation of reducing sugars. Hexose accumulation has been shown to be cultivar-dependent and proposed to be the result of sucrose hydrolysis via invertase. To study whether hexose accumulation is indeed related to the amount of invertase activities, two different approaches were used: (i) neutral and acidic invertase activities as well as soluble sugars were measured in cold-stored tubers of 24 potato cultivars differing in the cold-induced accumulation of reducing sugars and (ii) antisense potato plants with reduced soluble acid invertase activities were created and the soluble sugar accumulation in cold-stored tubers was studied. The cold-induced hexose accumulation in tubers from the different potato cultivars varied strongly (up to eightfold). Large differences were also detected with respect to soluble acid (50-fold) and neutral (5-fold) invertase activities among the different cultivars. Although there was almost no correlation between the total amount of invertase activity and the accumulation of reducing sugars there was a striking correlation between the hexose/sucrose ratio and the extractable soluble invertase activity. To exclude the possibility that other cultivar-specific features could account for the obtained results, the antisense approach was used to decrease the amount of soluble acid invertase activity in a uniform genetic background. To this end the cDNA of a cold-inducible soluble acid invertase (EMBL nucleic-acid database accession no. X70368) was cloned from the cultivar Desirée, and transgenic potato plants were created expressing this cDNA in the antisense orientation under control of the constitutive 35S cauliflower mosaic virus promotor. Analysis of the harvested and cold-stored tubers showed that inhibition of the soluble acid invertase activity leads to a decreased hexose and an increased sucrose content compared with controls. As was already found for the different potato cultivars the hexose/sucrose ratio decreased with decreasing invertase activities but the total amount of soluble sugars did not significantly change. From these data we conclude that invertases do not control the total amount of soluble sugars in cold-stored potato tubers but are involved in the regulation of the ratio of hexose to sucrose. Publication Types: Comparative Study Research Support, Non-U.S. Gov't PMID: 8580777 [PubMed - indexed for MEDLINE] 2334: J Toxicol Environ Health. 1996 Jan;47(1):1-30. Role of rat strain in the differential sensitivity to pharmaceutical agents and naturally occurring substances. Kacew S, Festing MF. Department of Pharmacology, University of Ottawa, Ontario, Canada. The development of drugs to combat diseases, chemicals to improve food production, or compounds to enhance the quality of life necessitates, by law, the use of laboratory animals to test their safety. In order to simulate the human condition it is necessary to choose a species in which pharmacokinetic and toxicokinetic mechanisms are established and resemble those of humans. The advantages of the use of the rat in drug and chemical toxicity testing include (a) metabolic pathway similarities to humans; (b) numerous similar anatomical and physiological characteristics; (c) a large database, which is extremely important for comparative purposes; and (d) the ease of breeding and maintenance of animals at relatively low cost. However, the choice of rat can be complicated, especially when over 200 different strains of rat are known to exist. The aim of this review is to summarize genetically determined differences in the responsiveness of rat strains to drugs and naturally occurring chemicals and to show that susceptibility is dependent on the target organ sensitivities, which may also be strain dependent. It is suggested that detailed studies of strain differences may help to clarify toxic mechanisms. Such studies are usually best conducted using inbred strains in which the genetic characteristics have been fixed, rather than in outbred stocks in which individual samples of animals may differ, the phenotype is variable, and the stocks are subject to substantial genetic drift. The fact that strains may differ also needs to be taken into account in assessing the potential hazard of the chemical, particularly when a study involves only a single strain and therefore provides no assessment of likely strain variation. Publication Types: Review PMID: 8568909 [PubMed - indexed for MEDLINE] 2335: Hum Reprod Update. 1995 Nov;1(6):523-42. Genetic engineering in plants. Simoens C, Van Montagu M. Laboratorium voor Genetica, Universiteit Gent, Belgium. Until now most research, and its funding, has been focused on animal and human health care as well as simple microbiological model systems such as Escherichia coli and yeast. Molecular plant studies have generally lagged behind, often simply adapting discoveries from the animal field to plants. Clearly, good health and the efficient tackling of diseases is crucial for the well-being of humans, and good remedies have a high economic value for the pharmaceutical industry. However, one should not forget that plants are an essential component of the large ecosystem that is our planet. They are not only the basic food producers but they are also necessary for a balanced atmosphere (oxygen production) and stable and viable climates. Especially in this period of demographic explosion and growing environmental deterioration, there is a need to rebuild our agricultural systems. Plants also have a wide variety of 'non-food' uses, for instance as energy sources, construction materials, or cosmetics. Last, but not least, they produce a lot of chemicals that can be used as pharmaceuticals. The growing awareness of the importance of plants has coincided with the development of plant molecular biology. Specific features make them ideally suited for gene engineering and genetic studies in general. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 9079394 [PubMed - indexed for MEDLINE] 2336: Biosci Biotechnol Biochem. 1995 Nov;59(11):2151-2. Assessment of the mutagenicity of extracts of TMV-coat-protein-gene induced transgenic tomato by the umu-test. Shinmoto H, Tomizawa A, Kobori M, Tsushida T, Shinohara K. National Food Research Institute, Ibaraki, Japan. We examined the mutagenicity of extracts (juice and ethanol extract) from a transgenic tomato that was established by transfection of a gene encoding the coat protein of tobacco mosaic virus (TMV) to the F1 hybrid between Lycopersicon esculentum LA1000 and L. peruvianum PI128650, by the umu-test with Salmonella typhimurium TA1535/pSK1002 as the test organism. The extracts showed no detectable mutagenicity. The extracts from the above-mentioned F1 hybrids and wild tomatoes and cultivars (L. peruvianum PI128650, L. peruvianum PI126944, L. pimpinellifolium LS1524, L. pimpinellifolium LA722, L. hirsutum LS503, Mini-carol, Sun-cherry, Momotaro, Odoriko, Kagome77, and Ponderosa) also showed no detectable mutagenicity. PMID: 8541659 [PubMed - indexed for MEDLINE] 2337: Invest Ophthalmol Vis Sci. 1995 Nov;36(12):2531-41. Macular pigment density in monozygotic twins. Hammond BR Jr, Fuld K, Curran-Celentano J. Schepens Eye Research Institute, Boston, Massachusetts 02114, USA. PURPOSE. Research shows wide variation in macular pigment density between individuals. As are other ocular pigments, this variation may be genetic. To test this hypothesis, the authors measured macular pigment density, serum carotenoid concentrations, and general dietary patterns in 10 pairs of identical twins. METHODS. Macular pigment was measured psychophysically by a 1 degree test stimulus. Foveal and parafoveal sensitivities to 460-nm and 530-nm light were compared. Determining the difference in log sensitivity to the 460-nm light for the fovea (where macular pigment is most dense) and the parafovea (where macular pigment is optically immeasurable), after normalizing with respect to 530 nm, yields a measurement of the optical density of macular pigment. Concentrations of carotenoids within the serum were measured using reverse-phase, high-performance liquid chromatography. Dietary patterns were determined using a food-frequency questionnaire. RESULTS. Statistically significant differences in macular pigment optical density were found for 5 of the 10 twin pairs. For these five pairs, differences in macular pigment density were moderately related to differences in the intake of dietary fat, iron, linoleic and oleic acid, fiber, and total calories (P < 0.10, individually; P < 0.05, for an equally weighted composite of these variables). There was no significant relationship, however, found between macular pigment density and carotenoids in the blood and diet. CONCLUSIONS. Given the putative protective role of macular pigment, variations in macular pigment density may have clinical significance. The conclusion that macular pigment is not completely determined genetically allows the possibility that macular pigment density may be modified for the protective purposes. The current data suggest that dietary fat, iron, and fiber may influence macular pigment levels (perhaps through their influence on carotenoid metabolism). These data suggest that the eventual deposition of macular pigment in the retina is complex and probably is influenced by a number of variables. Publication Types: Research Support, Non-U.S. Gov't Twin Study PMID: 7591643 [PubMed - indexed for MEDLINE] 2338: Trends Biotechnol. 1995 Sep;13(9):388-92. Transgenic plants as vaccine production systems. Mason HS, Arntzen CJ. Boyce Thompson Institute for Plant Research, Ithaca, NY 14853-1801, USA. Transgenic plants that express foreign proteins with industrial or pharmaceutical value represent an economical alternative to fermentation-based production systems. Specific vaccines have been produced in plants as a result of the transient or stable expression of foreign genes. It has recently been shown that genes encoding antigens of bacterial and viral pathogens can be expressed in plants in a form in which they retain native immunogenic properties. Transgenic potato tubers expressing a bacterial antigen stimulated humoral and mucosal immune responses when they were provided as food. These results provide 'proof of concept' for the use of plants as a vehicle to produce vaccines. Publication Types: Review PMID: 7546570 [PubMed - indexed for MEDLINE] 2339: Can Vet J. 1995 Aug;36(8):494-502. The pathogenesis and diagnosis of canine hip dysplasia: a review. Fries CL, Remedios AM. Department of Veterinary Anesthesiology, Radiology and Surgery, Western College of Veterinary Medicine, University of Saskatchewan, Saskatoon. Hip dysplasia is a common developmental problem affecting the canine population. Despite extensive research into the condition, many questions remain unanswered and numerous misconceptions are present among the general public. The purpose of this paper is to review the current knowledge on the development of hip dysplasia, factors modifying its development, and current diagnostic techniques.A computerized literature search was conducted for the period of January 1983 to April 1985 using the MEDLINE and CAB databases, and the keywords hip dysplasia, hip, dog, and canine. Other articles,wherever possible original research articles, published before 1983 were also reviewed.Animals affected by hip dysplasia are born with normal hips, but quickly develop subluxation of the femoral head. Degenerative joint disease follows.Hip dysplasia is a complex, inherited, polygenic trait. Selective breeding of only normal dogs with normal littermates, parents, and grandparents is there commended method of reducing the incidence in the general population.Gene expression in affected individuals may be modified by a number of environmental factors.These factors do not cause hip dysplasia, but they alter manifestations of the trait and its severity.Nutrition is a major environmental factor. Excess energy consumption increases the frequency and severity of hip dysplasia in genetically predisposed dogs. Food intake should be regulated to maintain a slender figure with the ribs and dorsal vertebral spines easily palpable, but not visible. Excess dietary calcium and vitamin D contribute to hip dysplasia in genetically predisposed individuals and should be avoided. High dose vitamin C supplementation ingrowing puppies does not prevent hip dysplasia, and this practice should be discontinued.Animals must be 2 years old before they can be certified as normal, but the disease may be diagnosed earlier. Earlier diagnosis of the condition would be very useful for the selection of breeding stock, but palpation techniques and the standard extended view radiographs have unacceptably high rates of error in young puppies. Stress radiography techniques may improve the accuracy of early diagnosis in the future. Publication Types: Review PMID: 7585436 [PubMed - indexed for MEDLINE] 2340: Mol Gen Genet. 1995 Jul 22;248(1):25-32. Nutritional response in a Drosophila yolk protein gene promoter. Søndergaard L, Mauchline D, Egetoft P, White N, Wulff P, Bownes M. Department of Genetics, University of Copenhagen, Denmark. The yolk protein genes (yps) of Drosophila melanogaster are only expressed in the ovary and fat body of female flies if they are supplied with proteinaceous food. This nutritional response is specific to the yp genes. We have used transgenic flies transformed with a series of constructs bearing deletions in the upstream region of the yp1 and yp2 genes attached to a reporter gene to search for DNA sequences responsible for the nutritional induction specific for yp1 and yp2 genes. Several regions were shown independently of each other to confer nutritional regulation on the expression of the yp1 and yp2 genes. This regulation can be induced both on the yp promoter and the heterologous Drosophila heat-shock 70 (hsp 70) promoter. The redundancy of sequences conferring a nutritional response on the yp genes is similar to that observed for the female specificity of these genes and suggest that several DNA binding proteins interact to provide the correct regulation of these genes. These results suggest that nutrition acts to modify the level of a trans-acting factor in the fat body. Northern blot analysis showed that the transcript levels from the dsx gene are not affected by nutrition, indicating that the response is not mediated via the dsx gene. Publication Types: Research Support, Non-U.S. Gov't PMID: 7651324 [PubMed - indexed for MEDLINE] 2341: Psychopharmacology (Berl). 1995 Jul;120(2):227-35. Effects of the dopamine D1 receptor antagonist SCH 39166 on the ingestive behaviour of alcohol-preferring rats. Panocka I, Ciccocioppo R, Mosca M, Polidori C, Massi M. Institute of Pharmacology, University of Camerino, Italy. The present study evaluated the effect of the selective and long-acting dopamine D1 receptor antagonist SCH 39166 on several aspects of the ingestive behaviour of genetically selected alcohol-preferring rats, bred from Sardinian alcohol-preferring rats. The effect of subchronic (8 days) subcutaneous drug administration was evaluated on the simultaneous daily intake of 10% ethanol, food and water. SCH 39166, 0.1 mg/kg, did not significantly modify the intake of the three ingesta. The dose of 1 mg/kg differentially modified rat ingestive behaviour, inhibiting intake of 10% ethanol, without modifying total fluid and food intake. The higher dose of 5 mg/kg produced a non-selective suppression of ingestive behaviour, which was accompanied by behavioural impairment. Acute drug injection was tested on 2-h intake of 10% sucrose, 0.1% saccharin, water or food. The doses of 0.1 and 1 mg/kg markedly inhibited the 2-h intake of 10% sucrose and 0.1% saccharin, but they did not modify either the 2-h water intake in water deprived and water sated rats or the 2-h food intake in food deprived rats. These findings suggest an important role of mechanisms mediated by D1 receptors in the control of alcohol and sweet solution intake, but not in water and food intake. Moreover, they indicate that SCH 39166, in relation to its selectivity and long-lasting activity, is an interesting pharmacological tool to investigate further the role of D1 receptor mechanisms in the control of ingestive behaviour. PMID: 7480557 [PubMed - indexed for MEDLINE] 2342: Poult Sci. 1995 May;74(5):852-63. Influence of the alv6 recombinant avian leukosis virus transgene on production traits and infection with avian tumor viruses in chickens. Gavora JS, Benkel B, Spencer JL, Gagnon C, Crittenden LB. Centre for Food and Animal Research, Agriculture and Agri-Food Canada, Ottawa, Ontario. The biological costs of the alv6 recombinant transgene that in chickens induces dominant resistance to the subgroup A avian leukosis virus (ALV), in terms of effects on production traits, were studied. Four generations of White Leghorn chickens of Line TR, segregating for alv6 but free of endogenous viral genes, as well as two generations of crosses between TR and Ottawa Line WG (WGTR) were tested under a specific-pathogen-free environment. In the birds studied, the transgene appeared unchanged compared to the original alv6: No major changes in alv6 DNA were detected by restriction analysis, the transgene did not express the group-specific antigen of ALV, and its presence was associated with absence of immune response to ALV. In most test years, and both TR and WGTR genomic backgrounds, alv6 was associated with delayed sexual maturity by 4 to 6 d, reduced egg production to 497 d of age by 20 to 46 eggs, and a 3.6 to 15% decline in egg production rate. No consistent effects on other traits, including mortality, were detected. When inoculated with the AC-1 isolate of Marek's disease virus in a separate experiment, TR birds with alv6 had a significantly lower body weight gain to 10 d of age than their sibs without the transgene. Thus, transgenesis has biological costs that have to be assessed against desirable effects of transgenes. Publication Types: Research Support, Non-U.S. Gov't PMID: 7603962 [PubMed - indexed for MEDLINE] 2343: J Environ Pathol Toxicol Oncol. 1995;14(3-4):133-57. Yeasts: from genetics to biotechnology. Russo S, Berkovitz Siman-Tov R, Poli G. Institute of Microbiology and Immunology, Faculty of Veterinary Medicine, University of Milan, Italy. Yeasts have been known and used in food and alcoholic fermentations ever since the Neolithic Age. In more recent times, on the basis of their peculiar features and history, yeasts have become very important experimental models in both microbiological and genetic research, as well as the main characters in many fermentative production processes. In the last 40 years, advances in molecular biology and genetic engineering have made possible not only the genetic selection of organisms, but also the genetic modification of some of them, especially the simplest of them, such as bacteria and yeasts. These discoveries have led to the availability of new yeast strains fit to fulfill requests of industrial production and fermentation. Moreover, genetically modified and transformed yeasts have been constructed that are able to produce large amounts of biologically active proteins and enzymes. Thus, recombinant yeasts make it easier to produce drugs, biologically active products, diagnostics, and vaccines, by inexpensive and relatively simple techniques. Yeasts are going to become more and more important in the "biotechnological revolution" by virtue of both their features and their very long and safe use in human nutrition and industry. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 9003692 [PubMed - indexed for MEDLINE] 2344: Methods Mol Biol. 1995;49:439-51. ELISA detection of foreign proteins. Mills EN, Plumb GW, Morgan MR. Department of Food Molecular Biochemistry, AFRC Institute of Food Research, Norwich, UK. PMID: 8563827 [PubMed - indexed for MEDLINE] 2345: Adv Exp Med Biol. 1995;369:221-8. Genetic engineering of foods to reduce the risk of heart disease and cancer. Knauf VC, Facciotti D. Calgene, Inc., Davis, California 95616, USA. Gene manipulation techniques can be used to increase, decrease, or add specific proteins to the edible parts of transgenic crop plants. With some basic understanding of plant biosynthetic pathways, then, the targeting of genes encoding specific enzymes allows the direct modification of the biochemical composition of foods. At Calgene, we have engineered the chemical composition of canola vegetable oils. Transgenic canola which are otherwise exactly like regular canola plants produce seed with oils a) that are modified in average fatty acid carbon chain length, b) that are modified in content of saturated fatty acids (both lower and higher), or c) that contain structured lipids. In principle, although the gene target may not be obvious, the relative amount of lipids compared to other nutrients can be decreased or increased in foods like peanut or soybean. The oil content of some food products might be modified to enhance levels of medium chain triglycerides or to contain "fish oil" fatty acids--without the cost or olfactory disadvantages. On a broader scope, the amino acid composition of proteins in basic grains is being pursued by several groups. Specific vitamin contents such as Vitamin A or E might be enhanced in basic foodstuffs; type and content of fiber may eventually be manipulated. Specific components such as caffeine or phytic acid conceivably can be eliminated in the source plant, negating the need for processing steps that add cost and that lessen flavor and nutrition. Clearly, the biochemical compositions of foods is complex and varied. Moreover, tailoring foods to better meet our needs is an expensive and lengthy process. The best possible understanding of how nutrition promotes good health is necessary to direct and prioritize our efforts to improve plants as food sources.(ABSTRACT TRUNCATED AT 250 WORDS) Publication Types: Review PMID: 7598011 [PubMed - indexed for MEDLINE] 2346: Methods Mol Biol. 1995;44:207-22. The plant oncogenes rolA, B, and C from Agrobacterium rhizogenes. Effects on morphology, development, and hormone metabolism. Michael T, Spena A. AFRC Institute of Food Research, Norwich Research Park, Colney, UK. Publication Types: Comparative Study PMID: 7581667 [PubMed - indexed for MEDLINE] 2347: Peptides. 1995;16(3):533-7. Subcutaneous injections of the tachykinin senktide reduce alcohol intake in alcohol-preferring rats. Ciccocioppo R, Panocka I, Pompei P, Polidori C, de Caro G, Massi M. Institute of Pharmacology, Faculty of Pharmacy, University of Camerino, Italy. The present study evaluated the effect of SC injections of the selective NK3 tachykinin agonist, Suc-[Asp6,MePhe8]substance P(6-11), also referred to as senktide (SENK), on 8% alcohol intake in genetically selected alcohol-preferring rats. Animals were offered access to 8% ethanol for 2 h/day (between 1800 and 2000 h) and to tap water for 4 h/day (between 1800 and 2200 h); SENK was injected 10 min before access to fluids. The peptide significantly reduced alcohol intake at doses of 125 and 250 micrograms/kg, but not at 62.5 micrograms/kg. The reduction in alcohol intake was accompanied by a sharp increase in water intake, so that total fluid intake was never significantly modified. The same SC doses of SENK did not modify water intake in rats with access to water, as the only fluid, for 4 h/day. In food-deprived rats food intake was not altered by 125 micrograms/kg, whereas 250 micrograms/kg produced a reduction in food intake that was smaller in intensity and shorter lasting than the reduction in alcohol intake. The same doses of SENK did not modify 0.1% saccharin intake, nor did they elicit major competing behaviors. The results of the present study are in keeping with those obtained following central injection of NK3 agonists, and show that a behaviorally selective reduction of alcohol intake can be evoked also by peripheral administration of SENK. Publication Types: Research Support, Non-U.S. Gov't PMID: 7544465 [PubMed - indexed for MEDLINE] 2348: Proc Natl Acad Sci U S A. 1994 Oct 25;91(22):10310-4. Disruption of virus movement confers broad-spectrum resistance against systemic infection by plant viruses with a triple gene block. Beck DL, Van Dolleweerd CJ, Lough TJ, Balmori E, Voot DM, Andersen MT, O'Brien IE, Forster RL. Molecular Genetics Group, Horticultural and Food Research Institute of New Zealand Ltd., Auckland. White clover mosaic virus strain O (WClMV-O), species of the Potexvirus genus, contains a set of three partially overlapping genes (the triple gene block) that encodes nonvirion proteins of 26 kDa, 13 kDa, and 7 kDa. These proteins are necessary for cell-to-cell movement in plants but not for replication. The WClMV-O 13-kDa gene was mutated (to 13*) in a region of the gene that is conserved in all viruses known to possess triple-gene-block proteins. All 10 13* transgenic lines of Nicotiana benthamiana designed to express the mutated movement protein were shown to be resistant to systemic infection by WClMV-O at 1 microgram of WClMV virions per ml, whereas all plants from susceptible control lines became systemically infected. Of the 13* transgenic lines, 3 selected for their abundant seed supply were shown to be resistant to systemic infection when challenged by inoculation with three different WClMV strains (O, M, and J) or with WClMV-O RNA at 10 micrograms/ml. Most plants were also resistant to systemic infection at inoculum concentrations up to 250 micrograms of WClMV virions per ml. In addition, the three 13* transgenic plant lines were found to be resistant to systemic infection with two other members of the Potexvirus group, potato virus X and narcissus mosaic virus, and the Carlavirus potato virus S but not to be resistant to tobacco mosaic virus of the Tobamovirus group. These results indicate that virus resistance can be engineered into transgenic plants by expression of dominant negative mutant forms of triple-gene-block movement proteins. PMID: 7937946 [PubMed - indexed for MEDLINE] 2349: Politics Life Sci. 1994 Aug;13(2):195-203. Transgenesis in farm animals: ethical implications for public policy. Mepham TB. Centre for Applied Bioethics, University of Nottingham, Sutton Bonington Campus, Loughborough LE12 5RD, UK. There is currently considerable investment in research aimed at producing transgenic farm animals with enhanced productive capacities. This article submits these prospective technologies to critical ethical evaluation. The analysis provided, focusing on issues relevant to public policy, suggests the need to introduce additional regulations governing the use of these technologies. Should their use be permitted in principle, specific legislation would seem to be required: (1) to protect the welfare of transgenic animals produced/kept in commercial enterprises; (2) to ensure the freedom of choice of purchasers of food and other products derived from transgenic animals by provision of adequate information on the source of the products; and (3) to provide a mechanism for limiting the use of transgenic technologies to those that are in the public interest. PMID: 15702517 [PubMed - indexed for MEDLINE] 2350: J Anim Sci. 1994 Jun;72(6):1641-53. Quantitative- and molecular-genetic effects on animal well-being: adaptive mechanisms. Newman S. Fort Keogh Livestock and Range Research Laboratory, ARS, Miles City, MT 59301. Domestic farm animals play an important role in meeting some basic needs of humankind, especially food and clothing. The aspects of genetic improvement programs in livestock production pertinent to animal welfare and animal well-being are reviewed. A link is made between the evolutionary processes of adaptation and domestication and animal well-being. Animal behavior is a component of all these. Thus, the genetics of behavior may provide clues to the well-being of farm animal populations, and it will also be of relevance to public opinion issues of animal welfare. Many expressions of behavior by domestic livestock may be influenced by those processes that change gene as well as genotypic frequencies such as inbreeding, drift, and artificial selection. The environment in which the individual lives will also play a role, along with the interaction between genotype and environment. Selection for or against such behaviors as aggressiveness, docility, response to stress, and certain sexual behaviors in some livestock species has often been successful. This points to the existence of additive genetic variation for behavior, and scope for the inclusion of behavioral traits into selection programs, if these measures are shown to be related to welfare. Negative relationships between behaviors associated with well-being and traits of economic importance have been reported in most livestock species. However, estimates of genetic parameters, especially genetic correlations between objective measures of well-being and production traits, are scarce. There have been no comprehensive studies of the welfare of transgenic animals reported in the scientific literature. Increased use of biotechnology in animal agriculture, coupled with greater public scrutiny of livestock industries, may precipitate decisions concerning the interface of behavior and genetics that need to be addressed before scientists can conduct appropriate experimental evaluations. Publication Types: Review PMID: 8071192 [PubMed - indexed for MEDLINE] 2351: Presse Med. 1994 Mar 26;23(12):558-60. [Etiology of Crohn disease. Current data] [Article in French] Colombel JF, Gower-Rousseau C. Both genetic and environmental factors appear to play an aetiologic role in Crohn's disease. The hypothesis of genetic susceptibility is based on the frequency of familial forms (6-33%) and higher relative risk in first degree relatives (x10 to x21). Three genetic segregation analyses have defined a genetic model favouring autosomal recessive transmission and incomplete penetration. The frequency of the deleterious allele would be 0.01 in the general population. One-third of all patients would be homozygous. To date, no genetic marker has been found for Crohn's disease although in one study, molecular biology techniques revealed a significant association between the HLA-DR1, DQW5 genotypes, and the association DR1/DQW5, and Crohn's disease. There are also many arguments in favour of an environmental cause, including: increased incidence since the Second World War, a north-south incidence gradient (established in the US and probable in Europe), predominance in urban areas. In addition, 21 conjugal forms were recently reported in the US, including 14 which appeared after marriage. We have also observed 10 conjugal forms in a limited area in northern France and in Belgium. Smoking has also been shown to have a detrimental effect, perhaps via modifications in the microcirculation of the intestinal wall. Despite contradictory results, oestrogen-progesterone treatment does not appear to increase the risk of Crohn's disease and there is no convincing evidence that any particular food could be incriminated. In contradiction with a widely believed myth, there is no evidence showing that psychiatric factors have an effect on appearance or aggravation of the disease. Perinatal infection has however been recently shown to affect incidence and subjects born during periods of flu epidemics have a higher relative risk. Still other studies have emphasized the dual role of genetic/environmental factors. Based on current knowledge, the sequence leading to the appearance of Crohn's disease would include one or more perinatal event(s) such as viral infection acting on a genetically susceptible subject and leading to modified immune response in a vulnerable system (perinatal period). Later in life an aggression (environment) would lead to inappropriate immune response. Current research is directed towards identifying infectious agents capable of triggering the disease and on markers of genetic susceptibility. Publication Types: Editorial English Abstract Review PMID: 8066056 [PubMed - indexed for MEDLINE] 2352: Genetics. 1994 Mar;136(3):1025-38. Unusual properties of regulatory DNA from the Drosophila engrailed gene: three "pairing-sensitive" sites within a 1.6-kb region. Kassis JA. Laboratory of Developmental Biology, Food and Drug Administration, Bethesda, Maryland 20892. We have previously shown that a 2-kb fragment of engrailed DNA can suppress expression of a linked marker gene, white, in the P element vector CaSpeR. This suppression is dependent on the presence of two copies of engrailed DNA-containing P elements (P[en]) in proximity in the Drosophila genome (either in cis or in trans). In this study, the 2-kb fragment was dissected and found to contain three fragments of DNA which could mediate white suppression [called "pairing-sensitive sites" (PS)]. A PS site was also identified in regulatory DNA from the Drosophila escargot gene. The eye colors of six different P[en] insertions in the escargot gene suggest an interaction between P[en]-encoded and genome-encoded PS sites. I hypothesize that white gene expression from P[en] is repressed by the formation of a protein complex which is initiated at the engrailed PS sites and also requires interactions with flanking genomic DNA. Genes were sought which influence the function of PS sites. Mutations in some Polycomb and trithorax group genes were found to affect the eye color from some P[en] insertion sites. However, different mutations affected expression from different P[en] insertion sites and no one mutation was found to affect expression from all P[en] insertion sites examined. These results suggest that white expression from P[en] is not directly regulated by members of the Polycomb and trithorax group genes, but in some cases can be influenced by them. I propose that engrailed PS sites normally act to promote interactions between distantly located engrailed regulatory sites and the engrailed promoter. PMID: 8005412 [PubMed - indexed for MEDLINE] 2353: Biotechnology (N Y). 1994 Mar;12(3):272-5. Accumulation of fructose polymers in transgenic tobacco. Ebskamp MJ, van der Meer IM, Spronk BA, Weisbeek PJ, Smeekens SC. Department of Molecular Cell Biology, University of Utrecht, The Netherlands. Fructan, a polyfructose molecule, is a storage compound in a limited number of plant species. Usually these species accumulate fructan with a low degree of polymerization (DP) and most of these plants have properties which preclude their use as a fructan source. With the eventual aim of allowing the accumulation of high DP fructans in non-fructan storing plants, we have investigated whether carbohydrate flow in the plant cell can be directed to produce this polymer. For this purpose the SacB gene from Bacillus subtilis, which encodes levansucrase, was modified and introduced into tobacco plants. Transgenic plants containing the sacB gene accumulate fructans. The size and properties of this fructan are similar to fructan produced by Bacillus subtilis, and is stable in plants. Although the level of fructan accumulation in the transgenic tobacco plants ranged from 3-8 percent of the dry weight, no levansucrase mRNA or protein could be detected in these plants. Extension of this work should permit the production of this high molecular weight biopolymer in crop plants for applications in food and non-food products. Publication Types: Research Support, Non-U.S. Gov't PMID: 7764488 [PubMed - indexed for MEDLINE] 2354: Transgenic Res. 1994 Jan;3(1):3-12. Opportunities for manipulating the seed protein composition of wheat and barley in order to improve quality. Shewry PR, Tatham AS, Halford NG, Barker JH, Hannappel U, Gallois P, Thomas M, Kreis M. Department of Agricultural Sciences, University of Bristol, AFRC Institute of Arable Crops Research, Long Ashton, UK. Wheat and barley are the major temperate cereals, being used for food, feed and industrial raw material. However, in all cases the quality may be limited by the amount, composition and properties of the grain storage proteins. We describe how a combination of biochemical and molecular studies has led to an understanding of the molecular basis for breadmaking quality in wheat and feed quality in barley, and also provided genes encoding key proteins that determine quality. The control of expression of these genes has been studied in transgenic tobacco plants and by transient expression in cereal protoplasts, providing the basis for the production of transgenic cereals with improved quality characteristics. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 8142950 [PubMed - indexed for MEDLINE] 2355: Arch Pediatr. 1994 Jan;1(1):5-10. [Nutrition, a model of interaction between genetic and environmental factors] [Article in French] Rey J, Bresson JL, Abadie V. Publication Types: Editorial Review PMID: 8087221 [PubMed - indexed for MEDLINE] 2356: Microb Releases. 1993 Dec;2(3):121-5. Gene transfer from engineered Lactococcus lactis strains to Enterococcus faecalis in the digestive tract of gnotobiotic mice. Gruzza M, Langella P, Duval-Iflah Y, Ducluzeau R. Unité d'Ecologie et Physiologie du Système Digestive, Institut National de la Recherche Agronomique, Jouy-en-Josas, France. The introduction of genetically modified organisms into food products requires an evaluation of the behaviour and the dissemination of foreign genes of such organisms among the human intestinal microflora. The conjugal transfer, both in vitro and in vivo (in mice digestive tract) of DNA from Lactococcus lactis donor strains to an Enterococcus faecalis strain isolated from human faecal flora was studied. We followed the transfer of (1) the self-transmissible plasmid pIL205; (2) two non-self-transmissible but mobilizable plasmids, pIL252 and pIL253; (3) one plasmid, pMS1.5B, integrated into the chromosome of L. lactis. In vitro, the transfer frequency of pIL205 (expressed as the number of transconjugants per donor cell) was 9.6 x 10(-4); mobilization of one of the non-self-transmissible plasmids, pIL253, was observed (4.9 x 10(-7)). In vivo, only transfer of pIL205 and pIL253 occurred, but the frequency was not determined. The transfer of pMS1.5B was not detected in vitro or in vivo. Publication Types: Research Support, Non-U.S. Gov't PMID: 8111532 [PubMed - indexed for MEDLINE] 2357: Philos Trans R Soc Lond B Biol Sci. 1993 Nov 29;342(1301):287-91. Commercialization of genetically engineered crops. Horsch RB. Monsanto Company, St Louis, Missouri 63198. More abundant harvests from insect- and disease-resistant crops, vine-ripened tomatoes, or less oily potato chips or french fries are some of the benefits that will result from single gene improvements under development today. These single gene traits will be combined with the best new varieties produced by traditional plant breeding and will accelerate the pace and the scope of our ability to develop even better and more productive crops in the future. The initial group of genetic improvements were first field tested in 1987, improved upon over the past 6 years, and are finally approaching the first commercial sales over the next 3 to 4 years. The key hurdles from discovery of a promising lead to a commercial product trait include: (i) gene cloning and expression; (ii) product development; (iii) field testing; (iv) breeding into multiple elite varieties; (v) product characterization and regulatory review; (vi) public acceptance; and (vii) marketing. The expense and risk to bring transformed crops to market successfully is significantly higher than for traditionally developed new varieties. The high value of some single gene targets and the possibility for patent protection of the processes and final products provide the incentive for private investment in this area. The value to farmers, consumers, the environment and society in general is very high because the problems being solved are those that have resisted previous attempts through conventional means. Public investment in basic plant science research and private investment in product development is a powerful combination for continual improvement in lowering the cost and improving the quality of the world's food supply. PMID: 8115451 [PubMed - indexed for MEDLINE] 2358: Hum Exp Toxicol. 1993 Nov;12(6):516-21. Implications of animal welfare on toxicity testing. Meyer O. National Food Agency of Denmark, Søborg. The testing strategy for chemical substances is discussed with regard to obtaining improved quality of data for health assessment while respecting the ethical responsibility for consideration of the welfare of the animals involved. Ensuring animal welfare without indulging too much in anthropomorphism leads to better research/testing. Current trends in toxicity testing will result in tests involving more sophisticated techniques, better quality of laboratory animals, and eventually the use of fewer animals. PMID: 7904468 [PubMed - indexed for MEDLINE] 2359: Genome. 1993 Oct;36(5):954-61. Expression of an amylase--alcohol dehydrogenase chimeric gene in transgenic strains of Drosophila melanogaster. Grunder AA, Loverre-Chyurlia A, Hickey DA. Centre for Food and Animal Research, Agriculture Canada, Ottawa, ON. A chimeric gene, consisting of 428 bp of the promoter sequences of the alpha-amylase gene of Drosophila melanogaster, fused to the transcribed region of the alcohol dehydrogenase (Adh) gene, was introduced into the genome of an Adhnull stock of Drosophila via P element mediated transformation. DNA analysis (Southern blotting) of three transformant strains confirmed the insertion of either one or two copies of the chimeric gene per strain. A histochemical study of ADH enzyme activity in dissected tissues of the transgenic larvae revealed that the chimeric Amy-Adh gene was expressed only in the posterior larval midgut and that this expression was repressed by dietary glucose, thus representing an expression pattern characteristic of the Amy gene. This indicates that the Amy upstream promoter sequences contain signals mediating both tissue specificity and glucose repression of the Adh structural gene in the transgenic larvae. The level of ADH activity expressed in transgenic flies was relatively low. This was paralleled by a low level of Adh mRNA, indicating a reduction in the transcriptional rate of the chimeric gene. Publication Types: Research Support, Non-U.S. Gov't PMID: 7505762 [PubMed - indexed for MEDLINE] 2360: J Exp Anim Sci. 1993 Sep;35(5-6):221-31. Multiple levels of response in carcinogenicity bioassays: regulational variation among viable yellow (Avy/-) mice. Wolff GL. Division of Nutritional Toxicology, National Center for Toxicological Research, Food and Drug Administration, Jefferson, Arkansas. Within genetically identical inbred and F1 hybrid test animal populations there exist subpopulations with different levels of sensitivity to induction of toxic endpoints, e.g., neoplasms, in response to toxicant exposure. These subpopulations differ from each other by other phenotypic characteristics as well. Presumably, these differences reflect alterations in the quantitative expression of some genes. These alterations are most probably directly or indirectly induced by subtle prenatal, neonatal or postnatal changes in endogenous microenvironmental conditions or factors. Such subpopulations have been identified within a population of agouti A/a and mottled yellow Avy/A (C3H x VY)F1 hybrid male mice. In these subpopulations differential body weight gain and formation of multiple liver adenomas in response to phenobarbital treatment were correlated with altered constitutive and inducible hepatic drug-metabolizing isozyme activities. These subpopulations could be identified by body weight as early as weaning age. In a different population of (YS x VY)F1 hybrid female mice, three phenotypes with different patterns of sensitivity to liver and lung tumor formation form visually separable phenotypic subpopulations by postnatal day 7. Two of these phenotypes, obese mottled yellow and lean pseudoagouti, are genetically identical (genotype: Avy/a), while the third phenotype, lean black a/a, differs from the others by just the Avy allele. The yellow and pseudoagouti mice, fed lindane (gamma-hexachlorocyclohexane) for 24 months, differed with respect to liver tumor incidence but had similar incidences of lung tumors. In contrast, the black mice, fed lindane, were totally resistant to both types of tumors. Analyses of phenotypic variation within other inbred or F1 hybrid populations have identified analogous subpopulations. In carcinogenicity assays this phenotypic variability can be used to mimic differential sensitivity to toxicant exposure among individuals in human populations. Data obtained from such phenotypic subpopulations should assist in improving risk assessment efforts. Publication Types: Review PMID: 8218437 [PubMed - indexed for MEDLINE] 2361: Plant Mol Biol. 1993 Jul;22(4):589-602. Identification and genetic analysis of normal and mutant phytoene synthase genes of tomato by sequencing, complementation and co-suppression. Fray RG, Grierson D. Department of Physiology and Environmental Science, University of Nottingham, Faculty of Agricultural and Food Science, Sutton Bonington, Loughborough, UK. A tomato phytoene synthase gene, Psy1, has recently been isolated as the clone GTOM5 and shown by sequence identity to be the gene from which the major fruit-ripening cDNA clone TOM5 was derived. Sequence analysis of transcripts from two allelic yellow-fruited tomato mutants, mapped to chromosome 3, has shown the lack of carotenoids in fruit of these mutants to be due to the production of aberrant TOM5 transcripts which are unlikely to encode a functional phytoene synthase enzyme. In one mutant (yellow flesh) the aberrant transcript contained a sequence that, by its strong hybridization to a wide size range of genomic fragments, appeared to be repeated many times within the genome. Southern and PCR analysis of the phytoene synthase genes in the mutant revealed restriction fragment length polymorphisms, suggesting that the production of altered mRNAs was associated with specific genomic rearrangements. Constitutive over-expression of a TOM5 cDNA clone in transgenic mutant plants restored synthesis of the carotenoid lycopene in ripening fruit and also led to unscheduled pigment production in other cell types. In some mutant plants transformed with the TOM5 cDNA construct, inhibition of carotenoid production in immature green fruit, leaves and flowers was observed, due to the phenomenon of co-suppression, indicating that different insertion events with the same gene construct can lead to overexpression or co-suppression in transgenic plants. Green organs of these plants were susceptible to photobleaching, due to the lack of carotenoids. These results suggest the existence of separate Psy genes for carotenoid synthesis in green organs. Publication Types: Comparative Study PMID: 8343597 [PubMed - indexed for MEDLINE] 2362: Eur J Pharmacol. 1993 Jun 15;246(1):67-71. Metformin and brown adipose tissue thermogenetic activity in genetically obese Zucker rats. Rouru J, Isaksson K, Santti E, Huupponen R, Koulu M. Department of Pharmacology, University of Turku, Finland. The effect of chronic metformin treatment on brown adipose tissue thermogenetic activity was investigated in young genetically obese Zucker rats. The binding of [3H]GDP to brown adipose tissue mitochondria, expression of uncoupling protein mRNA in brown adipose tissue, weight gains and cumulative food intakes were measured in metformin (320 mg/kg orally for 12 days)-treated obese Zucker rats as well as in pair-fed--and in ad libitum--fed control obese rats. The weight gains were identically reduced in the metformin- and pair-fed control group compared to the ad libitum--fed rats. Metformin also significantly reduced cumulative food intake. The binding of [3H]GDP to brown adipose tissue mitochondria and the expression of uncoupling protein mRNA in brown adipose tissue were not modified by metformin. It is concluded that the weight gain reducing effect of metformin in obese Zucker rats is mainly due to reduced food intake and does not involve an effect of metformin on brown adipose tissue thermogenetic activity. Publication Types: Research Support, Non-U.S. Gov't PMID: 8354343 [PubMed - indexed for MEDLINE] 2363: Br Poult Sci. 1993 May;34(2):383-91. Comparative utilisation of sulphur-containing amino acids by genetically lean or fat chickens. Leclercq B, Chagneau AM, Cochard T, Hamzaoui S, Larbier M. Station de Recherches Avicoles, INRA, Nouzilly, France. 1. Genetically lean (LL) or fat (FL) male chickens were fed from 28 to 47 days of age on 5 experimental diets differing by their methionine+cystine content (5.4, 5.8, 6.2, 6.6 and 7.0 g/kg, respectively). 2. Growth rate of LL chickens was reduced by the lower sulphur-containing amino acid (SAA) concentrations whereas that of FL was not modified. 3. LL chickens exhibited a larger feather protein gain than FL, which was stimulated by SAA intake. 4. SAA retention, when plotted against SAA consumption, was always greater in LL than in FL. 5. Large differences were observed between genotypes for plasma-free amino acids. Lysine, glutamic acid, histidine and serine were found at significantly higher concentrations in LL birds. Branched amino acids, aromatic amino acids, SAA and arginine were found at higher concentrations in FL. No differences were observed for aspartic acid, glycine, alanine and total amino acids. Methionine supplementation decreased free amino acid concentrations, with the exceptions of arginine and leucine. 6. It is concluded that lean chickens require a higher dietary concentration of SAA than FL. This is mainly caused by their lower food consumption and their greater feather synthesis. However, LL use SAA more efficiently than FL. Publication Types: Comparative Study PMID: 8513412 [PubMed - indexed for MEDLINE] 2364: New Sci. 1993 Apr 24;138(1870):26-31. Engineering the therapies of tomorrow. Coghlan A. PMID: 11656235 [PubMed - indexed for MEDLINE] 2365: J Anim Sci. 1993;71 Suppl 3:41-2. Food safety and inspection service update on food safety of animals derived from biotechnology experiments. Basu P, Masters B, Patel B, Urban O. United States Department of Agriculture, Food Safety and Inspection Service, Washington, DC. Recent progress in the field of biotechnology and the production of transgenic livestock has raised a question regarding the need for the regulation of these animals. There is also the need to regulate nontransgenic animals resulting from transgenic animal research. It is anticipated that several governmental agencies will be involved in regulatory issues pertaining to these animals. The United States Department of Agriculture (USDA), Food Safety and Inspection Service (FSIS) will ultimately be responsible for ensuring that transgenic animals intended for human consumption are wholesome, unadulterated, and properly labeled. The FSIS has implemented a program for the regulation of slaughtering nontransgenic animals resulting from transgenic animal experiments. However, the FSIS has not yet approved any transgenic livestock for slaughter. Scientists from the FSIS, in conjunction with other government agencies, are currently developing guidelines for the slaughter of transgenic animals. PMID: 8505269 [PubMed - indexed for MEDLINE] 2366: J Anim Sci. 1993;71 Suppl 3:43-6. The food safety of transgenic animals: implications from traditional breeding. Berkowitz DB. Office of Biotechnology HF-6, Food and Drug Administration, Rockville, MD 20857. The genetic events associated with traditional selection have implications for the food safety of transgenic animals. Selection has been empirical, relying on the use of the best animals for breeding. Molecular techniques are now being used to identify the genes selected and to describe the differences between alleles that are important in selection to improve quantitative traits. The results of such analyses provide background details of the genetic and physiological effects of the traditional selection of animal lines. Examples of the kinds of genes that may be subject to selection are those coding for peptide hormones, steroid metabolic enzymes, the calcium-channel gating protein, and genes of the major histocompatibility complex. Unselected genes, sometimes with undesirable alleles, may be carried along as "hitchhikers" if they are closely linked to the selected gene. In spite of this potential for physiologically dangerous genetic changes in selected animals, hereditary food toxicity has never been associated with a selected line of the common food animals. This is probably because the allowable physiological range of results of selection is limited by the requirement for healthy, productive animals. Based on these limitations, foods from healthy transgenic animals produced for the purpose of herd improvement are likely to be as safe as the foods from the untransformed parental line. Animals are important indicators of their own food safety. Publication Types: Review PMID: 8099351 [PubMed - indexed for MEDLINE] 2367: Biotechnol Genet Eng Rev. 1993;11:31-56. Biotechnology in aquaculture, with special reference to transgenic salmon. Male R, Lorens JB, Nerland AH, Slinde E. Center of Biotechnology, Norwegian Food Research Institute. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 7840846 [PubMed - indexed for MEDLINE] 2368: Australas Biotechnol. 1992 Dec;2(6):355-60. Genetic manipulation of milk proteins and its consequences for the dairy industry. Boland MJ, Hill JP, Creamer LK. Protein Chemistry Section, New Zealand Dairy Research Institute, Palmerston North. Genetic selection of cattle by selective breeding patterns dates back to prehistoric times and has resulted in the diversity of breeds we see today. Selection in New Zealand has been for fat production earlier in the century, and more recently for protein production as well as fat. There is a lot of interest today in the naturally occurring variants of the milk proteins, as these can confer interesting differences in the molecular behaviour of the proteins as well as being correlated with compositional differences in the milk. Genetic modification holds great promise for the future in the dairy industry, but present constraints due to cost, lack of basic knowledge, and difficulty in producing genetically-modified calves, mean that only the biopharmaceutical area is likely to be affected in the near future. Coupled to this is an apparent lack of acceptance of food from genetically-modified animals by consumers. It will therefore need a change in public attitude as well as some development in science and technology before dairy products from genetically modified cattle become a commercial reality. Publication Types: Review PMID: 1369112 [PubMed - indexed for MEDLINE] 2369: New Sci. 1992 Nov 14;136(1847):13-4. Guess what's coming to dinner? Vines G. Publication Types: News Personal Name as Subject: Rifkin J PMID: 11656186 [PubMed - indexed for MEDLINE] 2370: New Sci. 1992 Oct 3;135(1841):5. Danes wary about engineered animals. Radin P. Publication Types: News PMID: 11656161 [PubMed - indexed for MEDLINE] 2371: Mol Mar Biol Biotechnol. 1992 Aug-Oct;1(4-5):376-9. Fish gonadotropin-releasing hormone gene and molecular approaches for control of sexual maturation: development of a transgenic fish model. Aleström P, Kisen G, Klungland H, Andersen O. Department of Food and Dairy Industries, Agricultural University of Norway. The prepro-GnRH gene and mRNA primary structure were fully established from Atlantic salmon (Salmo salar) and partially from rainbow trout (Oncorhynchus mykiss). Results show that the GnRH coding region of 30 base pairs is well conserved during evolution. In contrast, the GnRH-associated peptide (GAP) sequence shows very limited homology when the GnRH genes from mammalian and teleost species are compared. A simple method for selecting transgenic fish after transfer of the firefly luciferase gene was developed. The method involves bioluminescent measurement of live animals in a scintillation counter. Publication Types: Research Support, Non-U.S. Gov't PMID: 1308825 [PubMed - indexed for MEDLINE] 2372: J Am Vet Med Assoc. 1992 Jul 15;201(2):228-34. Biotechnology and food safety. Kopchick JJ. Edison Animal Biotechnology Center, Ohio University, Athens 45701. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 1500314 [PubMed - indexed for MEDLINE] 2373: Science. 1992 Jun 26;256(5065):1747-9, 1832. Comment in: Science. 1992 Dec 4;258(5088):1561-2. The safety of foods developed by biotechnology. Kessler DA, Taylor MR, Maryanski JH, Flamm EL, Kahl LS. Food and Drug Administration (FDA), Rockville, MD 20857. PMID: 1615315 [PubMed - indexed for MEDLINE] 2374: Australas Biotechnol. 1992 Jun;2(3):152-66. Ten reactions to the government inquiry report. A summary. [No authors listed] PMID: 1368373 [PubMed - indexed for MEDLINE] 2375: Rural Sociol. 1992 Winter;57(4):476-93. Public opposition to genetic engineering. Hoban T, Woodrum E, Czaja R. PMID: 11654067 [PubMed - indexed for MEDLINE] 2376: Adv Food Nutr Res. 1992;36:89-208. Plant food protein engineering. Utsumi S. Research Institute for Food Science, Kyoto University, Japan. Publication Types: Research Support, Non-U.S. Gov't Review PMID: 1497851 [PubMed - indexed for MEDLINE] 2377: Free Radic Res Commun. 1992;17(1):73-85. Stress resistance of Drosophila transgenic for bovine CuZn superoxide dismutase. Reveillaud I, Kongpachith A, Park R, Fleming JE. Linus Pauling Institute of Science and Medicine, Palo Alto, CA 94306. Several oxidative and non-oxidative stresses were applied to two transgenic strains of Drosophila melanogaster (designated P(bSOD)5 and P(bSOD)11) that express superoxide dismutase (SOD) at elevated levels, and control strains that express normal SOD levels. Transgenic strain P(bSOD)5 exposed to paraquat (1,1'-dimethyl-4,4'-bipyridinium dichloride), a redox cycling agent that generates superoxide anion when metabolized in vivo, was significantly more resistant to this xenobiotic than control flies. When test flies were subjected to 100% oxygen for 20 min each day, the mean lifespan was 3.62 days for control strain 25, but 4.35 days for both transgenic strains. The mortality curves of strains fed 1% H2O2 were similar, but the median lifespan of 72 h for controls and 64 h for transgenics suggests that the transgenic flies were slightly more sensitive to H2O2. The activity of catalase was the same for all strains. Using starvation resistance as a non-oxidative stress, flies maintained on water without any food had identical survival curves; for all strains, the median lifespan was 72 h. Throughout the lifespan, no statistically significant difference in physical activity was displayed for transgenic versus control flies. Collectively, these data suggest that the increased lifespan previously observed in SOD transgenics is specifically related to resistance to oxidative stresses. Publication Types: Research Support, Non-U.S. Gov't PMID: 1332918 [PubMed - indexed for MEDLINE] 2378: J Am Vet Med Assoc. 1991 Dec 15;199(12):1714-21. Implications of biotechnology, risk assessment, and communications for the safety of foods of animal origin. Acuff GR, Albanese RA, Batt CA, Berndt DL, Byers FM, Dale BE, Denton JH, Fuchs RL, Gastel B, Heidelbaugh ND, et al. College of Agriculture and Life Sciences, Texas A&M University, College Station 77843. Publication Types: Congresses PMID: 1687575 [PubMed - indexed for MEDLINE] 2379: J Endocrinol Invest. 1991 Dec;14(11):919-25. The effects of glucose ingestion and fasting on plasma immunoreactive beta-endorphin, adrenocorticotropic hormone and cortisol in obese subjects. Balon-Perin S, Kolanowski J, Berbinschi A, Franchimont P, Ketelslegers JM. Division of Endocrinology and Nutrition, University of Louvain Medical School (UCL), Brussels, Belgium. It has been demonstrated that opioid peptides are involved in the stimulation of food intake in rats and that the circulating beta-endorphin levels are increased in genetically obese rodents. Therefore, to assess whether the changes in food intake may influence circulating beta-endorphin levels in obese subjects, plasma beta-endorphin, ACTH and cortisol concentrations were determined in obese patients after an oral glucose load and during a 7-day total starvation. Baseline plasma beta-endorphin concentrations were significantly higher in obese patients than in control normal-weight subjects, while ACTH and cortisol levels were similar in both groups. Plasma beta-endorphin, ACTH and cortisol concentrations were not affected by the ingestion of 75 g glucose, neither were plasma beta-endorphin concentrations modified during prolonged starvation. Moreover, the lack of nycthemeral variations in beta-endorphin levels, documented before and during starvation while plasma ACTH and cortisol were significantly reduced in the evening, suggests that some extra anterior pituitary sources or some obesity-related changes in beta-endorphin metabolism may contribute to the pool of circulating beta-endorphin in obese subjects. On the other hand, even the extreme changes in nutritional conditions, such as total food deprivation or glucose ingestion, are devoid of any detectable influence on circulating beta-endorphin levels. PMID: 1666898 [PubMed - indexed for MEDLINE] 2380: Biotechnol Bioeng. 1991 May;37(11):1076-86. Kinetic analysis of the effects of plasmid multimerization on segregational instability of CoIE1 type plasmids in Escherichia coli B/r. Kim BG, Shuler ML. Department of Food Science and Technology, Cornell University, Ithaca, New York 14853. The effect of plasmid multimerization on segregational instability was investigated using a structured, segregated model of genetically modified Escherichia coli cells. By including the multimerization of plasmids, the model can predict the proportion of each multimer in the total plasmid population. Simulation results suggest that the plasmid copy number is controlled by the total plasmid content (i.e., total number of plasmid origins) in the host cell and that multimerization reduces the total number of independent, monomeric segregation units. However, multimerization is found to have a minor effect on decreasing plasmid segregational stability for multicopy plasmids with average copy number per cell greater than about 25. Also model predictions were used to test whether or not a nonrandom plasmid distribution at cell fission could cause segregational instability. Even in the case of severely biased partitioning, plasmids whose copy number is above 45 per cell do not show significant segregational instability. The results suggest that when the ColE1-type plasmid does not encode and express any large or disruptive foreign proteins, the copy number of 45 per cell may be the threshold at which only growth rate-dependent instability is responsible for overall plasmid instability. PMID: 18597339 [PubMed - in process] 2381: JAMA. 1991 Mar 20;265(11):1429-36. Comment in: JAMA. 1991 Jul 17;266(3):362-3. Biotechnology and the American agricultural industry. Council on Scientific Affairs, American Medical Association. [No authors listed] To meet the needs of a rapidly growing population and minimize the toxic influences of traditional farming practices on the environment, the American agricultural industry has applied molecular technology to the development of food crops and livestock. By placing genes specific for highly desirable phenotypes into the DNA of plants, animals, and bacteria, farmers have increased crop and livestock survival, enhanced the nutritional quality of foods, increased industry productivity, and reduced the need for toxic pesticides and herbicides. However, introduction of genetically modified foods into the marketplace has raised a spectrum of public health issues. Physicians, as the most proximal scientific resource for most individuals, are uniquely positioned to address patient concerns regarding the safety of genetically altered foods. This report provides an overview of the inherent risks and benefits of "agrogenetics" and offers a series of recommendations designed to promote the education of the medical community and dispel public misconception regarding genetic manipulation. KIE: The application of molecular technology to agriculture and animal husbandry and the introduction of genetically modified foods into the marketplace have raised questions in the minds of regulators and consumers. Physicians, as the scientifically-trained individuals most accessible to the general public, are in a position to influence the acceptance of genetically altered foods. This report summarizes the risks and benefits of agricultural genetics and offers five recommendations concerning American Medical Association activities that will enable physicians to educate the public and government officials about the benefits of agricultural biotechnology. A glossary of terms concludes the report. Publication Types: Review PMID: 1999885 [PubMed - indexed for MEDLINE] 2382: Growth Dev Aging. 1991 Summer;55(2):81-9. Body weight and longevity in genetically obese and non-obese mice fed fat-modified diets. Smith BA, Edwards MS, Ballachey BE, Cramer DA, Sutherland TM. Department of Food Science and Human Nutrition, Colorado State University, Fort Collins 80523. Polygenically obese and genetically related non-obese male mice were fed ad libitum purified diets differing in quantity or type of fat from weaning onward to determine effects upon weight gain and life span. Non-obese mice lived 71% longer than mice in the obese line (mean = 753 d versus 441 d). Obese mice fed a low-fat diet containing 1% corn oil by weight lived 26% longer than their obese counterparts fed high fat diets containing 20% additional fat as either corn oil or beef tallow. The low fat diet had more of a normalizing effect upon longevity than upon body weight in polygenically obese mice. Conversely, dietary fat concentration had little effect upon longevity in non-obese mice and more effect upon body weight than in genetically obese mice. Type of dietary fat had little effect upon body weight and no effect upon longevity. The results of this study suggest that life span was only partially explained by body weight, and furthermore, that genetics play a greater role than body weight or dietary fat concentration in determining life span. Publication Types: Research Support, U.S. Gov't, P.H.S. PMID: 1938048 [PubMed - indexed for MEDLINE] 2383: Trends Biotechnol. 1991 Jan;9(1):5-7. The proof of the cloning is in the eating. Geisow M. Biodigm, East Bridgeford, Nottingham, UK. Publication Types: Review PMID: 1366925 [PubMed - indexed for MEDLINE] 2384: Proc Nutr Soc. 1990 Oct;49(3):459-66. Growth promotion in farm animals. Buttery PJ, Dawson JM. Department of Applied Biochemistry and Food Science, University of Nottingham, School of Agriculture, Loughborough. Publication Types: Review PMID: 1981934 [PubMed - indexed for MEDLINE] 2385: Biotechnol Bioeng. 1990 Sep;36(6):581-92. A structured, segregated model for genetically modified Escherichia coli cells and its use for prediction of plasmid stability. Kim BG, Shuler ML. Department of Food Science and Technology, Cornell University, Ithaca, New York 14853. A structured, segregated model is presented for an asynchronously growing population of genetically modified Escherichia coli cells. A finite representation method was modified so that 272 cells could be used to represent a microbial population. The concept of a "limbo" compartment was introduced to allow random plasmid distribution to daughter cells upon cell division while restricting the number of computer cells included in the calculation. This scheme enabled us to predict plasmid instability and distribution of plasmid-originated properties in a population without a priori determination of growth rates or probability of forming plasmid-free cells from plasmid-containing cells. Predictions of population behavior using a single-cell model requires no adjustable parameters. The results comparing different induction strategies suggest that in continuous culture, there exists an optimum efficiency of partial induction that maximizes the long-term productivity of the gene product due to plasmid stability. With the optimum efficiency of partial induction, constant induction appears to prove more stable than cycling induction. PMID: 18595116 [PubMed - in process] 2386: Biotechnology (N Y). 1990 Sep;8(9):819-25. The food safety of transgenic animals. Berkowitz DB. Food Safety and Inspection Service, U.S. Department of Agriculture, Washington, DC 20250. PMID: 1366792 [PubMed - indexed for MEDLINE] 2387: Protein Eng. 1990 Aug;3(8):725-31. Improvement of nutritional value and functional properties of soybean glycinin by protein engineering. Kim CS, Kamiya S, Sato T, Utsumi S, Kito M. Research Institute for Food Science, Kyoto University, Japan. Glycinin is one of the predominant storage proteins of soybean. To improve its functional properties (heat-induced gelation and emulsification) and/or nutritional value, the A1aB1b proglycinin subunit was modified on the basis of genetically variable domains suggested from the comparison of amino acid sequences of glycinin-type globulins from various legumes and nonlegumes and the relationships between the structure and the functional properties of glycinin. Thus, nucleotide sequences corresponding to each of the variable domains were deleted from the cDNA encoding the A1aB1b proglycinin, and a synthetic DNA encoding four continuous methionines was inserted into the cDNA region corresponding to each of the variable domains. Expression plasmids carrying the modified cDNAs were constructed and expressed in Escherichia coli strain JM105. Some of the modified proteins were accumulated as soluble proteins in the cells at a high level and self-assembled. They exhibited functional properties superior to those of the native glycinin from soybean, which establishes the possibility of creating theoretically designed novel glycinins with high food qualities. Publication Types: Research Support, Non-U.S. Gov't PMID: 2217146 [PubMed - indexed for MEDLINE] 2388: Tijdschr Diergeneeskd. 1990 Jun 15;115(12):570-4. [Transgenesis; applications, welfare and ethics] [Article in Dutch] de Boer HA. Rijksuniversiteit Leiden, Faculteit der Wiskunde en Natuurwetenschappen. Biomedical proteins and proteins having a nutritional function can be produced in large quantities (as well as cheap) in the mammary gland of mammals such as cows. Several of these proteins are of major economic importance and will result in several new uses for milk. For example, milk of genetically modified cows will be produced especially for branches of the pharmaceutical industry or for the baby food industry. Production of these new proteins will not have any injurious effects on the animal as proteins in the mammary gland are characterised by the fact that they follow a one-way course. Publication Types: English Abstract PMID: 2368085 [PubMed - indexed for MEDLINE] 2389: Nutr Cancer. 1990;14(1):15-26. Enhancement of immune status by high levels of dietary vitamin B-6 without growth inhibition of human malignant melanoma in athymic nude mice. Gebhard KJ, Gridley DS, Stickney DR, Shulz TD. Department of Nutrition, School of Public Health, Loma Linda University, CA 92350. The effects of dietary vitamin B-6 supplementation on the development of human malignant melanoma (M21-HPB) xenografts and on in vitro responses of leukocytes were examined. Male athymic nude mice, five weeks old, were divided into two groups of 48 each and fed 20% casein diets containing pyridoxine (PN) at 4.1 (control diet) and 61.6 mg/kg diet for 10 weeks. After four weeks of dietary treatment, 20 animals from each dietary group were injected subcutaneously with 3 x 10(7) melanoma cells. After 4, 8, and 10 weeks of dietary regimen, animals from each group were killed and blood, liver, and spleen samples were obtained. Food consumption and mouse body weights were similar between groups, and no difference was noted in tumor incidence or volume. Noninjected and tumor-bearing mice given the PN 61.6 diet generally exhibited greater oxygen radical production by phagocytic cells from blood and spleen than did animals fed the PN 4.1 diet. Spleen and blood B lymphocyte proliferation in response to lipopolysaccharide (LPS) was enhanced (10 and 30%) in the noninjected animals given the PN 61.6 diet. In addition, tumor-bearing mice fed the PN 61.6 diet had significantly greater LPS-induced spleen cell proliferation at eight weeks when compared with mice consuming the PN 4.1 diet. Despite immune enhancement, tumor incidence and progression was not modified by a high level of dietary vitamin B-6. Therefore, it is tempting to speculate that tumor inhibition by high dietary vitamin B-6 may be mediated by T lymphocyte-dependent mechanisms that are lacking in these genetically immuno-deficient mice. Publication Types: Research Support, Non-U.S. Gov't PMID: 2367233 [PubMed - indexed for MEDLINE] 2390: Psychopharmacology (Berl). 1990;101(2):178-86. Free-choice responding for ethanol versus water in alcohol preferring (P) and unselected Wistar rats is differentially modified by naloxone, bromocriptine, and methysergide. Weiss F, Mitchiner M, Bloom FE, Koob GF. Research Institute of Scripps Clinic, Department of Neuropharmacology, La Jolla, CA 92037. The role of opioids, dopamine and serotonin in ethanol (EtOH) reward and preference was investigated in non-deprived, Alcohol-Preferring (P), and genetically heterogenous Wistar rats. Operant responding for ethanol was initiated using sweet-solution substitution procedures. The rats were then trained in 30-min daily sessions to respond for ethanol (10% v/v) versus water under a two-lever, free-choice contingency. All testing was conducted in the absence of water and food deprivation or addition of sweeteners to the ethanol drinking solution. Rats of both strains developed stable preferences in responding for ethanol over water and consumed ethanol at quantities sufficient to produce pharmacologically relevant mean blood alcohol concentrations (P-Rats: 98 +/- 19.6 mg%; unselected Wistars: 41.7 +/- 8.5 mg%). In P-rats, systemic naloxone (NAL; 0.125, 0.25 and 0.5 mg/kg) pretreatments resulted in a dose-dependent suppression in responding for both ethanol and water, but did not alter ethanol preference (expressed as percent ethanol of total intake). In contrast, bromocriptine (BRO; 1.0, 2.0 and 4.0 mg/kg) produced a significant, dose-dependent shift in preference from ethanol toward water by inhibiting responding for ethanol while enhancing water consumption. In unselected Wistar rats, NAL and BRO treatments produced changes in ethanol preference patterns similar to those observed in P-rats. However, compared to P-rats, these changes were smaller and not consistently dose dependent. No changes in ethanol preference and water or ethanol intake were observed with methysergide (MET; 2.5, 5.0, 10.0 mg/kg) in either strain of rat. Together, the results suggest a possible involvement of dopaminergic mechanisms in the reinforcing properties of ethanol.(ABSTRACT TRUNCATED AT 250 WORDS) Publication Types: Comparative Study Research Support, U.S. Gov't, P.H.S. PMID: 2349359 [PubMed - indexed for MEDLINE] 2391: Domest Anim Endocrinol. 1990 Jan;7(1):1-18. Characterization of transgenic livestock production. Pinkert CA, Dyer TJ, Kooyman DL, Kiehm DJ. DNX Incorporated, Animal Biology Research Center, Athens, OH 45701. The objective of transgenic livestock improvement projects is to develop and bring to market superior breeding stock, as well as germplasm for the artificial insemination and embryo transfer industries. Livestock animal biotechnology programs hold the promise of achieving, in a single generation, improvements in commercially important livestock species previously possible only through long-term traditional selective breeding practices or by chance mutation. Transgenic farm animals harboring growth hormone or metabolically related structural genes have been created. Studies of these animals demonstrate the effects of inadequate regulation of transgene expression. Research continues to explore the intricacies of developmental regulation of such genes and phenotypic consequences of mammalian gene transfer. Ultimately, genetically engineered livestock will provide producers with the benefit of increased production efficiencies while the consumer will have healthier animal food products. Conceivably, products will be produced with lower levels of fat, cholesterol, feed additives and pharmaceutical residues from animals with altered carcass composition that will result in greater nutritional benefit for the consumer. Publication Types: Review PMID: 2178860 [PubMed - indexed for MEDLINE] 2392: Plant Physiol. 1989 Nov;91(3):1020-1024. Inheritance and Expression of the Mouse Metallothionein Gene in Tobacco: Impact on Cd Tolerance and Tissue Cd Distribution in Seedlings. Maiti IB, Wagner GJ, Yeargan R, Hunt AG. Plant Physiology/Biochemistry/Molecular Biology Program, Department of Agronomy, University of Kentucky, Lexington, Kentucky 40546-0091. Genetically engineered seedlings obtained from self-fertilized transgenic tobacco (Nicotiana tabacum) contained and expressed the mouse metallothionein and kanamycin resistance marker genes and were more tolerant to cadmium stress than untransformed controls. Cadmium accumulation in leaves of transgenic seedlings exposed to a low, field-like Cd concentration (0.02 micromolar) was about 20% lower than that in untransformed controls. Genetic analysis of R1 and R2 progeny showed inheritance of the marker gene to be as a dominant Mendelian trait. These results suggest the possibility of developing transgenic plants with modified tolerance to heavy metal stress and food crops having lower Cd content. PMID: 16667104 [PubMed - as supplied by publisher] 2393: Drake Law Rev. 1988-1989;38(3):471-550. The biotechnology revolution and its regulatory evolution. Hoffman DE. University of Maryland School of Law, Baltimore, MD, USA. PMID: 16086466 [PubMed - indexed for MEDLINE] 2394: J Nutr. 1987 Mar;117(3):428-35. In vivo lipogenesis of genetically lean and fat chickens: effects of nutritional state and dietary fat. Saadoun A, Leclercq B. In vivo lipogenesis was estimated in liver and carcass of male chickens selected for leanness (LL) or fatness (FL) by use of tritiated water. Effects of nutritional state and of a high fat diet (90 g/kg) were examined. Different classes of liver lipids were also measured. Fed birds exhibited enhanced hepatic lipogenesis, which was more pronounced in FL than in LL birds (+73%, mean of three experiments). Extrahepatic lipogenesis was poorly influenced by nutritional state in LL birds and not modified in FL ones. The high fat diet induced a fall in liver triacylglycerol and reduced de novo lipogenesis more drastically in liver than in carcass of both lines. Reduction of carcass lipogenesis was less pronounced in FL than in LL. Starvation led to a fall in liver triacylglycerols, nonesterified fatty acids and free cholesterol. The fat diet had similar but less pronounced effects. In most circumstances FL chickens exhibited a higher liver triacylglycerol content than LL ones. Comparison of fed and refed birds 100 min after the beginning of refeeding showed that de novo lipogenesis did not reach a plateau and differences between lines were still not significant. Publication Types: Comparative Study PMID: 3572556 [PubMed - indexed for MEDLINE] 2395: Pharmacol Ther. 1987;35(1-2):163-215. Adaptive changes in thermoregulation and their neuropharmacological basis. Brück K, Zeisberger E. Justus-Liebig-University of Giessen, F.R.G. Adaptive changes of the thermoregulatory system include morphological and functional modifications. The morphological modifications such as changes in body shape and insulation need time periods of months to years to develop, unless they are genetically fixed and appear seasonally. In general, they are preceded by functional modifications, including changes in capacity of the effector systems and changes in regulatory characteristics, which need much less time to develop. These early changes in regulatory characteristics, which can be defined as deviations in threshold and gain of the thermoregulatory responses, have been described and subdivided into short-term (minutes) and long-term (weeks) modifications. Evidence for the participation of monoaminergic brain stem systems in these modifications has been reviewed. On the basis of recent insights into the organization of the thermoregulatory system, and of evaluation of experimental evidence from electrophysiological, neuropharmacological, and neuroanatomical studies it can be concluded that these systems are involved in adaptive modifications. Receiving information from several sensory systems they seem to deliver additional modulatory signals, which may interfere with the processing of specific thermal information at several sites. Theoretically, the central monoamines may participate in the control of thermal input, in the central integration of thermal signals, and in modification of output signals to thermoregulatory effectors. Best documented is their modulatory action on thermosensitive and thermointegrative hypothalamic neurons. There, the monoamines 5-hydroxytryptamine and noradrenaline act as antagonists, which enhance or diminish the effects of thermal afferents mediated by other transmitters. Moreover, the antagonistic monoaminergic systems are interconnected and can influence each other at the level of lower brain stem. The activity in central monoaminergic systems can also be modified by neurohumoral feedback mechanisms from the periphery. By means of these interrelations the vegetative responses of the organism can be corrected and optimized. These interrelations can explain also some cross-adaptive changes in the thermoregulatory threshold for shivering evoked by nonthermal factors such as food intake or long-distance running. Publication Types: Review PMID: 3321099 [PubMed - indexed for MEDLINE] 2396: Ann Allergy. 1984 Dec;53(6 Pt 2):643-8. Enzymatic maturation of the gastrointestinal tract and its relevance to food allergy and intolerance in infancy. McNeish AS. The biologic clock that determines the temporal sequence of maturation of digestive and absorptive processes in the gastrointestinal tract is genetically predetermined, but may be modified by dietary, hormonal, or other factors. In general it may be said that the gastrointestinal tract of full-term neonates is capable of digesting and absorbing a nutritionally adequate quantity of dietary protein but capacity is limited. Very low birth weight preterm infants, who are surviving the early neonatal period in increasing numbers, have immaturity of a wide range of digestive and membrane-associated absorptive processes; in addition macromolecular absorption may be increased. Whether a limited capacity to digest food protein results in increased or altered antigenic stimulation of these immature infants remains speculative with present knowledge. Immaturity of intestinal lactase may lead to problems of lactose intolerance, but there is recent evidence that lactase activity may be inducible by milk feeding. Publication Types: Review PMID: 6439078 [PubMed - indexed for MEDLINE] 2397: Appl Environ Microbiol. 1981 Dec;42(6):944-950. Evidence for Plasmid Linkage of Restriction and Modification in Streptococcus cremoris KH. Sanders ME, Klaenhammer TR. Department of Food Science, North Carolina State University, Raleigh, North Carolina 27650. Restriction and modification have been demonstrated in Streptococcus cremoris KH cells when infected by Streptococcus lactis C2 phage (designated c2) at an efficiency of plating of 2 x 10. The growth of c2 phage through KH cells produces modified progeny phage capable of unrestricted growth on KH cells. The ability of single-colony isolates of S. cremoris KH cultures to restrict and modify c2 phage was found to be variable. From 2 to 6.5% of colonies isolated were partially deficient in restrictive capacity, permitting a greater plaquing ability by c2 phage of 1.8 to 2.9 log cycles. No completely restrictionless mutants were isolated from 1,000 colonies examined. Mutants were shown to be deficient in both restriction and modification capabilities of the same specificity. The frequent occurrence of a genotypic change that resulted in the loss of both restriction and modification capacities indicated the involvement of plasmid deoxyribonucleic acid in genetically determining this specific restriction and modification system. S. cremoris KH was found to harbor 11 plasmid molecules, with molecular weights (x10) estimated to be 50, 41, 24, 18, 10, 7.4, 3.3, 3.0, 2.8, 2.5, and 1.5. Of the 27 mutants examined, 25 were missing the 10-megadalton plasmid. This consistent plasmid difference among the majority of mutants isolated supports the involvement of this plasmid in restriction and modification. Plasmid linkage of restriction and modification systems provides a genetic mechanism for the rapid development of phage-sensitive starter cultures due to the inherent instability of extrachromosomal elements. PMID: 16345908 [PubMed - as supplied by publisher]