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. PartyzantГіw 57, 24-100 PuЕ‚awy, 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 UniversitГ¤t MГјnchen, 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 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] 4: 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. OcaГ±a 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] 5: 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] 6: 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. ClГ©ment, 92290 ChГўtenay Malabry, France; Bayer CropScience, 355 rue DostoГЇevski, 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] 7: 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] 8: 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] 9: 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] 10: 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] 11: 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] 12: 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] 13: 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] 14: 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] 15: 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] 16: 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, FeijГі TS, Boaventura GT. College of Medicine, Federal Fluminense University, NiterГіi, 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] 17: 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] 18: 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] 19: 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] 20: 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] 21: Food Chem Toxicol. 2008 Dec;46(12):3808-17. Epub 2008 Oct 8. Subchronic feeding study of high oleic acid soybeans (Event DP-3Г5423-1) 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-3Г5423-1 (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] 22: 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] 23: 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] 24: 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] 25: 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] 26: 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] 27: 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] 28: 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] 29: 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] 30: 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, GullГ¬ 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] 31: 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] 32: 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), CharitГ© 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] 33: 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] 34: 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] 35: 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, NaroditskiД 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] 36: 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] 37: 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] 38: 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] 39: 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 BaГ±os, 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] 40: 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, SepГєlveda Bravo O, Ruiz Fuentes D, Denegri Coria M. Departamento de ProducciГіn 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 (AraucanГa 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 AraucanГa Region, but a significant proportion accepts genetic modification in food (50.7%). Publication Types: English Abstract PMID: 18589572 [PubMed - indexed for MEDLINE] 41: 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] 42: 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] 43: 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 PГєblica, Universidad de Navarra, EspaГ±a. 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] 44: 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] 45: 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] 46: 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] 47: 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-TГ¤nikon 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] 48: 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, BermГєdez-HumarГЎn 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] 49: 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] 50: Food Chem Toxicol. 2008 Jun;46(6):2201-13. Epub 2008 Feb 29. Subchronic feeding study of herbicide-tolerant soybean DP-356Г43-5 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-356Г43-5 (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] 51: 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] 52: 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] 53: 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. BГёhn T, Primicerio R, Hessen DO, Traavik T. GenГёk-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] 54: Appetite. 2008 Jul;51(1):129-36. Epub 2008 Feb 7. Acceptance of genetically modified foods: the relation between technology and evaluation. TenbГјlt 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] 55: 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, PГ¶ting 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, KГ¤renlampi S, Kiss J, Kryspin-SГёrensen I, Kuiper H, Nes I, Panopoulos N, Perry J, PГ¶ting 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] 56: 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] 57: 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, SchГ¶n J, Martens H, Einspanier R. Department of Veterinary Biochemistry, Freie UniversitГ¤t 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] 58: 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] 59: Eur J Nutr. 2008 Mar;47(2):99-103. Epub 2008 Mar 4. Zeaxanthin is bioavailable from genetically modified zeaxanthin-rich potatoes. Bub A, MГ¶seneder 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] 60: 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] 61: 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. UnitГ© Etude du Polymorphisme des GГ©nomes VГ©gГ©taux (EPGV) UR1279, Centre National de GГ©notypage (CNG), 2 rue Gaston CrГ©mieux 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] 62: Rev Panam Salud Publica. 2008 Jan;23(1):52-8. [Academic production on food labeling in Brazil] [Article in Portuguese] CГўmara MC, Marinho CL, Guilam MC, Braga AM. FundaГ§ГЈo Oswaldo Cruz (Fiocruz), Centro de Estudos da SaГєde 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 "rГіtulo 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] 63: 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] 64: 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. RamГrez M, Ambrona J. Departamento de MicrobiologГa (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] 65: 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] 66: 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] 67: 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] 68: 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. MГ©decine interne, immunologie clinique et allergologique, HГґpital 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] 69: 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-DelГЁgue 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] 70: 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] 71: 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] 72: 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] 73: 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] 74: 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] 75: Med Pregl. 2007 May-Jun;60(5-6):295-8. [Diseases caused by viruses and toxins in biological warfare and bioterrorism] [Article in Serbian] BojiД‡ I, Vukadinov J, MiniД‡ S. Specijalisticka ordinacija Dr BojiД‡ 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] 76: 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] 77: 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] 78: 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] 79: 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] 80: 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] 81: 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] 82: 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] 83: 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] 84: 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] 85: 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] 86: 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] 87: 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] 88: 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] 89: 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] 90: 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., SchrГёder, 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., SchrГёder, 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; SchrГёder, 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] 91: 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, OberdГ¶rfer R, Samuels F. NestlГ© 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] 92: 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] 93: 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] 94: 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] 95: 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] 96: 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] 97: 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 InvestigaciГіn en BiologГa Celular y Molecular (CIBCM), Ciudad de la InvestigaciГіn, 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] 98: 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. MГјller 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] 99: 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] 100: 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 <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] 101: 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] GarcГa JE. Centro de EducaciГіn Ambiental de la Universidad Estatal a Distancia y Escuela de Biologia de la Universidad de Costa Rica, San JosГ©, 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] 102: 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] 103: 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] 104: 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] 105: 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 SaГєde 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] 106: 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] 107: 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] 108: 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 LiГЁge, Sart-Tilman, LiГЁge, Belgium. sdanyi@ulg.ac.be <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] 109: 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-UniversitГ 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] 110: 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] 111: 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. SГ©ralini 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 (MГјnster) 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] 112: 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] 113: 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 UniversitГ 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] 114: 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] 115: 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]