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DNA methods: critical review of innovative approaches.

The presence of ingredients derived from genetically modified organisms (GMOs) in food products in the market place is subject to a number of European regulations that stipulate which product consisting of or containing GMO-derived ingredients should be labeled as such. In order to maintain these labeling requirements, a variety of different GMO detection methods have been developed to screen for either the presence of DNA or protein derived from (approved) GM varieties. Recent incidents where unapproved GM varieties entered the European market show that more powerful GMO detection and identification methods will be needed to maintain European labeling requirements in an adequate, efficient, and cost-effective way. This report discusses the current state-of-the-art as well as future developments in GMO detection.

DNA, Plant↗

Analysis and interpretation of data from real-time PCR trace detection methods using quantitation of GM soya as a model system.

Recent years have seen an increased interest in DNA trace detection methods involved in many areas of bioanalytical research, such as quantitation of genetically modified (GM) ingredients in food products. There is little in the way of standardisation of data handling from these methods, and the data generated needs to be analysed appropriately if the results are to be interpreted correctly. This paper describes particular aspects of real-time PCR trace detection methods in order to increase the understanding of data generated using this bioanalytical technique. Using the specific example of GM soya detection and quantitation, it focuses on the production of calibration curves based on the mean and individual data values, the interpretation of correlation coefficients, regression techniques, and discusses suitable data analysis arising from simple and more complex experimental designs following transformation. By using the approaches outlined in this paper, more accurate analysis of data from real-time PCR and GM trace detection methods could be achieved.

Calibration↗

GM foods and the misperception of risk perception.

Public opposition to genetically modified (GM) food and crops is widely interpreted as the result of the public's misperception of the risks. With scientific assessment pointing to no unique risks from GM crops and foods, a strategy of accurate risk communication from trusted sources has been advocated. This is based on the assumption that the benefits of GM crops and foods are self-evident. Informed by the interpretation of some qualitative interviews with lay people, we use data from the Eurobarometer survey on biotechnology to explore the hypothesis that it is not so much the perception of risks as the absence of benefits that is the basis of the widespread rejection of GM foods and crops by the European public. Some respondents perceive both risks and benefits, and may be trading off these attributes along the lines of a rational choice model. However, for others, one attribute-benefit-appears to dominate their judgments: the lexicographic heuristic. For these respondents, their perception of risk is of limited importance in the formation of attitudes toward GM food and crops. The implication is that the absence of perceived benefits from GM foods and crops calls into question the relevance of risk communication strategies for bringing about change in public opinion.

Biotechnology↗

Distortion of genetically modified organism quantification in processed foods: influence of particle size compositions and heat-induced DNA degradation.

Milling fractions from conventional and transgenic corn were prepared at laboratory scale and used to study the influence of sample composition and heat-induced DNA degradation on the relative quantification of genetically modified organisms (GMO) in food products. Particle size distributions of the obtained fractions (coarse grits, regular grits, meal, and flour) were characterized using a laser diffraction system. The application of two DNA isolation protocols revealed a strong correlation between the degree of comminution of the milling fractions and the DNA yield in the extracts. Mixtures of milling fractions from conventional and transgenic material (1%) were prepared and analyzed via real-time polymerase chain reaction. Accurate quantification of the adjusted GMO content was only possible in mixtures containing conventional and transgenic material in the form of analogous milling fractions, whereas mixtures of fractions exhibiting different particle size distributions delivered significantly over- and underestimated GMO contents depending on their compositions. The process of heat-induced nucleic acid degradation was followed by applying two established quantitative assays showing differences between the lengths of the recombinant and reference target sequences (A, deltal(A) = -25 bp; B, deltal(B) = +16 bp; values related to the amplicon length of the reference gene). Data obtained by the application of method A resulted in underestimated recoveries of GMO contents in the samples of heat-treated products, reflecting the favored degradation of the longer target sequence used for the detection of the transgene. In contrast, data yielded by the application of method B resulted in increasingly overestimated recoveries of GMO contents. The results show how commonly used food technological processes may lead to distortions in the results of quantitative GMO analyses.

DNA, Plant↗

Assessment of usual dietary intake in population studies of gene-diet interaction.

AIMS: Dietary intake is a critical environmental exposure when considering the effect of many genetic factors on disease risk. However, dietary intake is a complex and changing measure that requires particular care in assessment. DATA SYNTHESIS: Although weighed diet records can theoretically provide the most accurate assessment of intake, they are usually not realistic in large population studies due to heavy respondent burden, likelihood of poor compliance, and the cost of data entry. Multiple 24-h dietary recalls can provide excellent detail, allowing for diverse dietary practices, but they are costly and require multiple contacts with participants. Food frequency questionnaires are the most cost-effective tool for assessing usual intake, particularly for micronutrients with high day-to-day variability. However, they have limitations for diverse populations and recent studies have questioned their ability to measure macronutrient intakes for assessing diet and disease relationships. CONCLUSION: At the present time, food frequencies remain the most cost-effective tool for large population studies. However, their limitations must be fully appreciated and demonstration of validity for nutrients of concern in the populations under study is essential. When macronutrients are of key interest, consideration should be given to the use of multiple recalls. Records may be used only in educated and compliant populations. Continued efforts to improve dietary assessment methodology must be investigated.

Attitude to Health↗

Biotechnology in the global agri-food system.

The advent of biotechnology presents fundamental challenges to the global agri-food industry. While the scientific base for agri-food production is being revolutionised, it is not clear if or how the technology will be used. Proponents of biotechnology and a large portion of agri-food policy makers around the world project a positive future in which technology overcomes food shortages, improves the environment, heals or eliminates disease and leads to a prosperous and healthy society. A smaller but significant array of policy makers, citizens and consumers fear that the technology will exacerbate food insecurity, threaten the environment, endanger human health and ultimately impoverish society itself. Although scientists and industry are convinced the fears are unfounded, it is not clear that our social institutions will be able to adapt, adopt and use the technology in a way that will satisfy society and improve social welfare.

Agriculture↗

Industrial dimensions of food allergy.

Serious attempts to estimate the impact of allergic reactions to foods on public health did not begin until the 1980s. Until about 15 years ago food allergy was considered a minor aspect of food safety. Two developments probably prompted a radical re-appraisal of that situation. The first was the apparently inexorable rise in the prevalence of atopic diseases, of which food allergy forms a part, with its possible consequences highlighted by some well-publicised severe reactions. The second was the growth of genetic modification technology, manifested by the commercialisation of transgenic crops. Each of these developments impacted on the food industry in distinct ways. On the one hand, consumers with food allergies had to be enabled to avoid specific allergens in products formulated with existing ingredients. Food manufacturers therefore had to identify those specific allergens down to trace amounts in all the ingredients forming the product and label or remove them. On the other hand, the introduction of products using ingredients from novel sources required an assessment of the allergenicity of these ingredients as an integral part of safety assurance. The approaches used by the food industry to protect existing consumers who have food allergies and those at potential risk of sensitisation from novel proteins will be illustrated, emphasising how they need to be built into every stage of the life cycle of a product.

Consumer Product Safety↗

[Impact of genetic modifications on infectious diseases].

Genetic engineering offers the theoretical possibility to transfer any natural or modified gene into any living organism. This generates new and diverse situations which may contribute to the spreading of infectious diseases or on the contrary to control them. Problems may theoretically come from uncontrolled genes providing resistance to antibiotics, from the activation of genomic retroviral sequences, from enhanced sensitivity of the organism to pathogens, as well as from the generation of mutated microorganisms with a higher pathogenecity. On the contrary, various genetic modifications may create organisms resistant to infectious diseases, generate safe and efficient recombinant vaccines, or provide patients with proteins which stimulate their defense mechanisms. The major impacts of genetic modifications in the development of infectious diseases or on the contrary in their eradication are analyzed in this article.

Animals↗

Transgenes for tea?

So far, no compelling scientific evidence has been found to suggest that the consumption of transgenic or genetically modified (GM) plants by animals or humans is more likely to cause harm than is the consumption of their conventional counterparts. Despite this lack of scientific evidence, the economic prospects for GM plants are probably limited in the short term and there is public opposition to the technology. Now is a good time to address several issues concerning GM plants, including the potential for transgenes to migrate from GM plants to gut microbes or to animal or human tissues, the consequences of consuming GM crops, either as fresh plants or as silage, and the problems caused by current legislation on GM labelling and beyond.

Animals↗

Public perceptions of transgenic animals.

The field of animal biotechnology has been rapidly expanding and the development of transgenic animals has been part of this research expansion. How the public perceives such developments is an important component of policy considerations. In general, biotechnology applications have been judged with evident hierarchies of acceptability. There appearto be hierarchies in terms of the type of organism being modified, the purpose of the application, the means to attain particular ends, and the nature of the benefits obtained. While general awareness of biotechnology and its specific applications remains low to moderate, this article presents data regarding public acceptance of a variety of applications. These range from the use of animals as disease models and as sources for tissues and organs, to the use of transgenic animals for disease control, for food, and for the production of pharmaceutical and industrial products. Case-by-case judgments are evident, but at the same time, the application of criteria such as the nature of the organism being modified, the animal welfare aspects and the ethical-moral concerns are additional criteria for public judgments. These findings are discussed in the context of their implications for public policy.

Animals↗

Unintended effects in genetically modified crops: revealed by metabolomics?

In Europe the commercialization of food derived from genetically modified plants has been slow because of the complex regulatory process and the concerns of consumers. Risk assessment is focused on potential adverse effects on humans and the environment, which could result from unintended effects of genetic modifications: unintended effects are connected to changes in metabolite levels in the plants. One of the major challenges is how to analyze the overall metabolite composition of GM plants in comparison to conventional cultivars, and one possible solution is offered by metabolomics. The ultimate aim of metabolomics is the identification and quantification of all small molecules in an organism; however, a single method enabling complete metabolome analysis does not exist. Given a comprehensive extraction method, a hierarchical strategy--starting with global fingerprinting and followed by complementary profiling attempts--is the most logical and economic approach to detect unintended effects in GM crops.

Consumer Product Safety↗

The relevance of gene transfer to the safety of food and feed derived from genetically modified (GM) plants.

In 2000, the thematic network ENTRANSFOOD was launched to assess four different topics that are all related to the testing or assessment of food containing or produced from genetically modified organisms (GMOs). Each of the topics was linked to a European Commission (EC)-funded large shared cost action (see http://www.entransfood.com). Since the exchange of genetic information through horizontal (lateral) gene transfer (HGT) might play a more important role, in quantity and quality, than hitherto imagined, a working group dealing with HGT in the context of food and feed safety was established. This working group was linked to the GMOBILITY project (GMOBILITY, 2003) and the results of the deliberations are laid down in this review paper. HGT is reviewed in relation to the potential risks of consuming food or feed derived from transgenic crops. First, the mechanisms for obtaining transgenic crops are described. Next, HGT mechanisms and its possible evolutionary role are described. The use of marker genes is presented in detail as a special case for genes that may pose a risk. Furthermore, the exposure to GMOs and in particular to genetically modified (GM) deoxyribonucleic acid (DNA) is discussed as part of the total risk assessment. The review finishes off with a number of conclusions related to GM food and feed safety. The aim of this paper is to provide a comprehensive overview to assist risk assessors as well as regulators and the general public in understanding the safety issues related to these mechanisms.

Animal Feed↗

Ethical issues in livestock cloning.

Although cloning may eventually become an important technology for livestock production, four ethical issues must be addressed before the practice becomes widespread. First, researchers must establish that the procedure is not detrimental to the health or well-being of affected animals. Second, animal research institutions should evaluate the net social benefits to livestock producers by weighing the benefits to producers against the opportunity cost of research capacity lost to biomedical projects. Third, scientists should consider the indirect effects of cloning research on the larger ethical issues surrounding human cloning. Finally, the market structure for products of cloned animals should protect individual choice, and should recognize that many individuals find the prospect of cloning (or consuming cloned animals) repugnant. Analysis of these four issues is complicated by spurious arguments alleging that cloning will have a negative impact on environment and genetic diversity.

Agriculture↗

Elliot Entis.

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Animals↗