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[Strategies for safety assessment of genetically modified crops: current and future development].

Gene recombinant technologies supply agriculture product with great vitality. But the risk of genetically modified crops cannot be ignored. The international organizations such as WHO, FAO and OECD have reached common agreement: the safety of transgenic crops should be thoroughly evaluated based on "substantial equivalence"principle. The relevant strategies including: substantial equivalent analysis, toxic tests, protein allergenic study, nutritional assessment, etc. With the development of new technologies, the approaches of genomic, proteomics, metabolomics would be applied to detect the unintended effects. The EU recently adopted legislation on the cultivation GM crops requiring the post market surveillance for any unanticipated adverse effects in the long term. In conclusion, the strategies of the safety assessment of GM crop are very strict and likely development.

Animals↗

DNA separation by capillary electrophoresis with hydrophilic substituted celluloses as coating and sieving polymers. Application to the analysis of genetically modified meals.

A coating procedure based on the physical adsorption of hydroxypropyl cellulose onto the wall of a capillary column has been successfully used for the separation of DNA fragments up to 500 bp. The method uses a running Tris-phosphate-EDTA buffer containing 2-hydroxyethyl cellulose as sieving polymer. The separation procedure shows good reproducibility (measured as RSD%) for consecutive runs (<0.64), for different days (< 1.15) and capillaries (<2.15), short analysis times, and a long coating lifetime. Good reproducibility and efficiency are even achieved by performing the separation in the presence of additives such as ethidium bromide and mannitol. The method is applied to the detection of GMOs in soybean and maize meals with an accurate evaluation of the length of DNA sequences, previously amplified by polymerase chain reaction.

Adsorption↗

How Japanese students reason about agricultural biotechnology.

Many have claimed that education of the ethical issues raised by biotechnology is essential in universities, but there is little knowledge of its effectiveness. The focus of this paper is to investigate how university students assess the information given in class to make their own value judgments and decisions relating to issues of agricultural biotechnology, especially over genetically modified organisms (GMOs). Analysis of homework reports related with agricultural biotechnology after identification of key concepts and ideas in each student report is presented. The ideas were sorted into different categories. The ideas were compared with those in the reading materials using the same categories. These categories included: concern about affects on humans, affects on the environment, developing countries and starvation, trust in industry, responsibility of scientists, risk perception, media influence, need for (international) organizations or third parties, and information dissemination. What was consistent through the different years was that more than half of the students took a "neutral" position. A report was scored as "neutral" when the report included both the positive and negative side of an issue, or when the student could not make a definite decision about the use of GMOs and GM food. While it may be more difficult to defend a strong ''for" or "against" position, some students used logical arguments successfully in doing so. Sample comments are presented to depict how Japanese students see agricultural technology, and how they value its application, with comparisons to the general social attitudes towards biotechnology.

Agriculture↗

Extraction and PCR analysis of soy DNA in chocolate.

The manner in which to discriminate between genetically modified (GM) versus non-modified foodstuffs is based on the presence of newly introduced genes at the protein or DNA level. Current available methods are almost exclusively based on the polymerase chain reaction (PCR). These methods consist of three steps: DNA isolation, amplification of the desired DNA fragment and visualisation of the obtained amplification products. The first and crucial step is the DNA isolation. In this study three different methods are described for the isolation of DNA from chocolate, two of which are commercial kits. The results indicate that both kits, in contrast with the non-commercial method, are suitable for the isolation of DNA from chocolate, provided that the adapted PCR conditions are applied.

Cacao↗

[Laboratory-performance study of the notified methods to detect genetically modified papaya (55-1)].

To investigate important factors affecting the reliability of the analytical results, proficiency tests were attempted for the histochemical method (GUS method) and the qualitative PCR method (PCR method) to detect genetically modified papaya (55-1) in the Japanease official method. The test samples were distributed to twenty-three laboratories that participated in the study and were examined according to the protocol. All the data collected from participating laboratories were statistically analyzed. In the PCR method, one negative sample was detected as positive using detection primers in one laboratory, though the sample was negative when checked using confirmation primers. Contamination might have occurred in the step of the preparation of the PCR sample solution using detection primers. In the GUS method, all the test samples were identified as expected. Thus, all the laboratories reported correct results overall.

Carica↗

Environmental biosafety and transgenic potato in a centre of diversity for this crop.

The Nuffield Council on Bioethics suggests that introgression of genetic material into related species in centres of crop biodiversity is an insufficient justification to bar the use of genetically modified crops in the developing world. They consider that a precautionary approach to forgo the possible benefits invokes the fallacy of thinking that doing nothing is itself without risk to the poor. Here we report findings relevant to this and other aspects of environmental biosafety for genetically modified potato in its main centre of biodiversity, the central Andes. We studied genetically modified potato clones that provide resistance to nematodes, principal pests of Andean potato crops. We show that there is no harm to many non-target organisms, but gene flow occurs to wild relatives growing near potato crops. If stable introgression were to result, the fitness of these wild species could be altered. We therefore transformed the male sterile cultivar Revolucion to provide a genetically modified nematode-resistant potato to evaluate the benefits that this provides until the possibility of stable introgression to wild relatives is determined. Thus, scientific progress is possible without compromise to the precautionary principle.

Agriculture↗

Multiplex polymerase chain reaction/membrane hybridization assay for detection of genetically modified organisms.

To improve detection efficiency and result accuracy, four screening primer pairs, four identifying primer pairs, one common primer pair and corresponding probes were designed for the development of multiplex polymerase chain reaction/membrane hybridization assay (MPCR-MHA) for detection of the foreign genes insert in genetically modified organisms (GMOs). After detecting condition and parameter were optimized and determined, MPCR reactions were developed for amplifying several target genes simultaneously in one tube. Primers were labeled with biotin at the 5'-end; biotinylated MPCR products were detected by hybridization to the oligonucleotide probes immobilized on a membrane with subsequent colorimetric detection to confirm hybridization. The testing of screening primers can judge whether the sample contains GMOs, and that of identifying primers can further judge what kinds of trait genes are contained in the sample. We detected nine soybean samples, six maize samples, seven potato samples and two rice samples by the MPCR-MHA method; at the same time we also detected them with single PCR-MHA method. The results between two methods have good consistency.

DNA Primers↗

[Opinions of a group of university students about science and technology].

OBJECTIVE: To learn the opinions of university students of four different areas on the impact of science and technology on society. SUBJECTS: One Hundred and sixty three close to graduate students of the Universidad Autonoma Metropolitana campus Iztapalapa, distributed as follows: Administration 59, Biology 50, Social Sciences 36 and Engineering 18. METHODS: For the survey we translated into spanish part of a questionnaire employed in several countries to explore ideas on the impact of science and technology on society of several groups. It contained general questions such as. Do you believe that science and technology are equally good or bad to society, or degree of knowledge of several technologies such as computation or in vitro fertilization. It includes also more specific questions, such as would your have problems with the use of genetically modified vegetables? RESULTS AND DISCUSSION: The results suggested that Administration and Social Sciences students had less interest in Science and Technology than the other, and that in general, the knowledge of all students is rather limited including biotechnology, genetic enginering and gene therapy. We compared the results with those obtained previously in a group of Mexican Physicians and Biology students from India, Thailand and Singapor.

Acquired Immunodeficiency Syndrome↗

Detection of genetically modified crops and their derivatives: critical steps in sample preparation and extraction.

The detection of genetically modified crops in foodstuff relies on detection of transgenic DNA or protein material in the sample matrix. Purified DNA or proteins are used as analytical material for polymerase chain reaction technologies and immunodiagnostics. Successful sample preparation is critical to the validity of subsequent analysis. For routine analysis, a good sample preparation technique should be simple, safe, and inexpensive while reproducibly generating DNA/protein of sufficient quality and yield. The suitability of isolated DNA or protein as an analyte for a detection or characterization technique depends on amount or concentration, purity, and integrity, each of which may be influenced by sample matrix and the extraction technique, and, in turn, may impact the validity of analytical techniques. The key sample preparation steps of homogenization, pretreatment, extraction, and purification are discussed as well as typical analytical methods. Consideration is given to application of these steps for particular sample matrixes to maximize yield, reduce inhibition effects, and minimize contamination. The choice of the most appropriate and valid methods for sample preparation from particular foods is discussed with respect to DNA analysis. Attention is also given to ease of use, cost, and generic applicability of the procedures.

Crops, Agricultural↗

Functional dynamics of living systems and genetic engineering.

The discussion on Genetically Modified Organisms (GMO's) has been centred mainly on the nature and effects on economy, human health, environment, of the few transgenic plant lines present in the market in the last eight years. On the contrary, the present paper starts with a discussion of some of the relevant changes in our basic knowledge of the structure and dynamics of living systems in the last twenty years. Contemporary Biology is then compared with what may be called the "modern paradigm" of life sciences on which present day GMO's are conceptually based. Technical, environmental, social and economic problems deriving from the unexpected, persistent prevalence of the old fashioned modern vision of life in the "spirit of time" will be thoroughly discussed with a particular attention to the virtualisation process of GMO's and the effects of the prevalence over economic, social, environmental reality of their symbolic values.

Animals↗

Sampling grain shipments to detect genetically modified seed.

Using the binomial distribution, the effect of sample size on the variability among sample test results when sampling a lot with 1.0% genetically modified (GM) or biotech seed was evaluated. The coefficient of variation, cv, among 500-seed sample test results taken from a lot with truly 1.0% was computed to be 44.5%. Increasing sample size to 1000 seeds reduced the cv among sample test results to 31.5%. The effects of sample size and accept/reject limits on the buyer's risk (bad lots accepted) and the seller's risk (good lots rejected) was also evaluated assuming a tolerance of 1.0% GM seed. Increasing sample size decreases both the buyer's and seller's risks at the same time. Using an accept/reject limit below the regulatory tolerance decreases the buyer's risk, but increases the seller's risk. Using an accept/reject limit above the regulatory tolerance decreases the seller's risk but increases the buyer's risk.

Analysis of Variance↗

Ultrastructural analysis of testes from mice fed on genetically modified soybean.

We have considered the possible effects of a diet containing genetically modified (GM) soybean on mouse testis. This organ, in fact, is a well known bioindicator and it has already been utilized, for instance, to monitor pollution by heavy metals. In this preliminary study, we have focussed our attention on Sertoli cells, spermatogonia and spermatocytes by means of immunoelectron microscopy. Our results point out that the immunolabelling for Sm antigen, hnRNPs, SC35 and RNA Polymerase II is decreased in 2 and 5 month-old GM-fed mice, and is restored to normal at 8 months. In GM-fed mice of all ages considered, the number of perichromatin granules is higher and the nuclear pore density lower. Moreover, we found enlargements in the smooth endoplasmic reticulum in GM-fed mice Sertoli cells. A possible role played by traces of the herbicide to which the soybean is resistant is discussed.

Animals↗

Regulatory and associated political issues with respect to Bt transgenic maize in the European union.

Legislation at the national level in Europe as well as that developed by the European Union (EU) generally permits release and commercialization of genetically modified organisms (GMOs). However, only 10 plant/event combinations were registered as of 2002: three maize events (Bt176, Mon810, and Bt11), with the other seven divided among carnation (3), oil-seed rape (2), tobacco (1), and raddiccio (1). Of these, only one maize event (Bt176) has been registered as a legal variety, and this was in Spain, where 22,000ha have been planted annually since 1998. In this paper, we first provide an overview on the complexity of EU GMO legislation. Then we discuss the minor role that results of EU-funded biosafety research have had on governmental policy. Finally, we provide information about initiatives for post-commercialization monitoring plans of Bt maize in Europe. As a result of the slow progress to date, we conclude that commercialization of GMOs will be seriously delayed in the EU for the next several years.

Animals↗

Coherence between legal requirements and approaches for detection of genetically modified organisms (GMOs) and their derived products.

Analytical methods for the qualitative and quantitative detection of genetically modified (GM) products may serve multiple purposes. Legal requirements differ among jurisdictions, ranging from no requirements to mandatory use of event-specific quantitation and implementation of production chain traceability. Although efforts have been taken to harmonize the analytical methodology at national, regional, and international levels, no normative international standards have yet been established. Lack of coherence between analytical methodologies and their applicabilities, on the one hand, and legislation, on the other hand, is a major problem. Here, key points where coherence is lacking are discussed. These include the definition of units of measurements, expression of GM material quantities, terminology, and inconsistent legal status of products derived from related but slightly different transformation routes. Finally, recommendations to improve the coherence are brought forward, including guidance to stakeholders for prediction of product-specific GM material quantities from gene ratios in the originating seed.

Base Sequence↗

In vitro screening of food peptides toxic for coeliac and other gluten-sensitive patients: a review.

Experience gained through investigations on coeliac disease makes it possible to propose a screening method based on agglutination of isolated K562(S) cells to evaluate the occurrence in food protein of amino acid sequences that are able to adversely affect coeliac and related gluten-sensitive patients. The method consists of in vitro sequential peptic and tryptic digestion of food protein fractions under optimal pH, temperature and time conditions and in vitro incubation of the digest with K562(S) cells; the toxic potential is detected as an agglutination of K 562 (S) cells after a short incubation. Other in vitro test systems, including atrophic coeliac intestinal mucosa and rat fetal intestine, can be used to confirm the results obtained with the isolated cells. A fractionation step of the proteolytic digest on a sepharose-mannan column before exposure of the in vitro systems to the separated peptide fractions adds to the sensitivity of the method. This screening method is not only very useful to investigate action mechanisms in coeliac disease, but also to assess the safety of genetically-modified plant foods and novel foods for gluten-sensitive patients.

Amino Acid Sequence↗

Effects of feeding calves genetically modified corn bt11: a clinico-biochemical study.

Genetically modified corn Bt11 is insect-resistant and expresses Cry1Ab toxin, an insecticidal protein, in kernels. Although Bt11 corn is considered safe based on animal performance, there are no reports available on the clinico-biochemical effects of feeding it to cattle. In this study, we evaluated the effects of feeding Bt11 to calves, using blood and ruminal clinico-biochemical parameters. Our three-month-long feeding experiment demonstrated that calves (n=6), fed with a ration containing 43.3% of Bt11 corn kernels as dry matter, did not develop any discernible clinical, hematological, biochemical, or ruminal abnormalities as compared with control calves (n=6) fed non-Bt11 corn. The results suggest that the transgenic Bt11 has no negative clinico-biochemical effects on calves.

Alanine Transaminase↗

Explaining public resistance to genetically modified corn: an analysis of the distribution of benefits and risks.

Genetically modified (GM) crops have met with widespread approval among scientists and policy makers in the United States, but public approval of GM crops, both domestically and abroad, is progressing much more slowly. An underlying cause of public wariness may be that both nations and individual consumers do not perceive significant benefits to themselves from GM crops, while fearing the risks they may incur. In this study, an economic analysis is conducted to determine whether the benefits of one type of GM corn, Bt corn (genetically modified to resist damage from the ECB and Southwestern corn borer), outweigh the potential risks; and who the "winners" and "losers" are among stakeholder groups that may be affected by Bt corn. It is found that Bt corn growers, consumers, and industry all benefit from Bt corn adoption, though the purported health and environmental benefits of reducing chemical pesticide usage through Bt corn are negligible. Though the aggregated public benefit is large, the welfare gain to individual consumers is small and may not make up for perceived risks. While environmental and health risks of Bt corn are unlikely, the potential market risks-impacting both the organic corn market and total U.S. corn exports-are found to be significant. Currently, distributional analysis is not a part of regulatory decision making of Bt corn in the United States; yet it may help to explain why decision makers at both the government and individual-consumer levels have failed to embrace Bt corn and other GM crops.

Food, Genetically Modified↗

A detection method of CryIAc protein for identifying genetically modified rice using the lateral flow strip assay.

We examined the lateral flow strip assay for identifying unauthorized genetically modified (GM) rice. The GM rice expresses the Bacillus thuringiensis (Bt) toxin, CryIAc protein, which confers tolerance to insects. The recombinant CryIAc protein was prepared from the inclusion bodies of an E. coli. strain into which the CryIAc gene had been inserted, using gel filtration chromatography. The lateral flow strip assay for the identification of GM cotton which also expresses CryIAc protein, was applied to unpolished rice and polished rice spiked with recombinant CryIAc protein. The spiked recombinant CryIAc protein was clearly detected at the level of 0.012 microg/g in both the unpolished and polished rice. After loading of the extract on the strip, a 60 -minute stand time is necessary to clearly detect CryIAc protein. The detection limit was approximately 12 ng CryIAc protein per gram of rice. These results suggest that the lateral flow strip assay for GM cotton can be used to detect CryIAc protein expressed in GM rice.

Bacillus thuringiensis Toxins↗