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PubMed2006

ELISA method for monitoring human serum IgE specific for Cry1Ab introduced into genetically modified corn.

Enzyme-linked immunosorbent assay (ELISA) is the most convenient method of monitoring the occurrence of IgE antibodies specific for novel proteins in genetically modified (GM) foods. The levels of IgE specific for a recombinant protein, Cry1Ab, were determined using an ELISA method. A soluble form of the Cry1Ab protein purified from pCold1 vector-transformed Escherichia coli pTf16/BL21 was used as the ELISA coating antigen, and 1M NaCl was used as the washing buffer to remove IgE non-specifically bound to the coated antigen. Sera from 44 patients allergic to major food allergens were obtained, diluted 20-fold, tested, and found no identifiable IgE above background levels. We also tested sera from patients with corn allergy against whole extracts of non-GM and GM-corn (MON 810) using immunoblotting. The staining patterns were similar for the two types of corn. These results indicate that significant levels of IgE antibodies specific to Cry1Ab were not found in the sera of Japanese patients with food allergies.

Bacillus thuringiensis Toxins↗
PubMed2003

Analysis of protein profiles of genetically modified potato tubers by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.

Traceability of genetically modified (GM) foods demands the development of appropriate reliable techniques in order to identify and quantify peptide or nucleic acid residues in GM plants and food products through the food chain. In this study the applicability of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) was demonstrated for the characterization of proteins of transformed and untransformed potato (Solanum Tuberosum L.) tubers. In GM tubers the expression level of the G1-1 gene, which regulates transition from dormancy to sprouting tubers, was inhibited by antisense technology. The analysis of antisense transformed lines showed that several of them exhibited a significant delay in sprouting relative to the control lines, in accordance with a decrease in the transcript level. Preliminary attempts to compare the protein patterns obtained from transformed and control lines using traditional electrophoresis were not able to reveal differences in the low-kDa range. Instead, MALDI-TOFMS applied to total peptide extract without any purification was able to distinguish spectral patterns of transformed and untransformed lines. In particular, several characteristic peaks from m/z 4373 to 4932 were detected only in the mass spectra of GM tuber samples.

Food, Genetically Modified↗
PubMed2004

Trust, the asymmetry principle, and the role of prior beliefs.

Within the risk literature there is an ongoing debate on whether trust is vulnerable or enduring. Previous research on nuclear energy by Slovic in 1993 has shown that negative events have much greater impact on self-reported trust than do positive events. Slovic attributes this to the asymmetry principle: specifically, that trust is much easier to destroy than to create. In a questionnaire survey concerning genetically modified (GM) food in Britain (n= 396) we similarly find that negative events have a greater impact on trust than positive events. Because public opinion in Britain is skewed in the direction of opposition toward GM food, the pattern of results could either be caused by the fact that negative information is more informative than positive information (a negativity bias) or reflect the influence of people's prior attitudes toward the issue (a confirmatory bias). The results were largely in line with the confirmatory bias hypothesis: participants with clear positive or negative beliefs interpreted events in line with their existing attitude position. However, for participants with intermediate attitudes, negative items still had greater impact than the positive. This latter finding suggests that, congruent with the negativity bias hypothesis, negative information may still be more informative than positive information for undecided people. The study also identified the labeling of GM products, consulting the public, making biotechnology companies liable for any damage, and making a test available to detect GM produce as being particularly important preconditions for maintaining trust in the regulation of agricultural biotechnology.

Attitude↗
PubMed2002

Ultrastructural morphometrical and immunocytochemical analyses of hepatocyte nuclei from mice fed on genetically modified soybean.

No direct evidence that genetically modified (GM) food may represent a possible danger for health has been reported so far; however, the scientific literature in this field is still quite poor. Therefore, we carried out an ultrastructural morphometrical and immunocytochemical study on hepatocytes from mice fed on GM soybean, in order to investigate eventual modifications of nuclear components of these cells involved in multiple metabolic pathways related to food processing. Our observations demonstrate significant modifications of some nuclear features in GM-fed mice. In particular, GM fed-mice show irregularly shaped nuclei, which generally represents an index of high metabolic rate, and a higher number of nuclear pores, suggestive of intense molecular trafficking. Moreover, the roundish nucleoli of control animals change in more irregular nucleoli with numerous small fibrillar centres and abundant dense fibrillar component in GM-fed mice, modifications typical of increased metabolic rate. Accordingly, nucleoplasmic (snRNPs and SC-35) and nucleolar (fibrillarin) splicing factors are more abundant in hepatocyte nuclei of GM-fed than in control mice. In conclusion, our data suggest that GM soybean intake can influence hepatocyte nuclear features in young and adult mice; however, the mechanisms responsible for such alterations remain unknown.

Aging↗
PubMed2005

Past challenges faced: an overview of current educational activities of IUTOX.

Over the past decade, educational programmes have been the main focus of the activities of the International Union of Toxicology (IUTOX). The IUTOX educational programmes are dynamic and have been growing in scope and frequency each year. It is envisaged that this growth will continue with guidance from our member societies and the continuing support of our sponsors. Presently, IUTOX is engaged in the following educational programmes: (1) International congresses that provide the opportunity for direct communication of current toxicological information. Fellowships are sponsored to facilitate attendance at these congresses for toxicologists in need. (2) Workshops that permit interaction on a more localised level of topics of more regional interest. Workshops have served to help stimulate formation of toxicology societies by bringing together sufficient scientists to facilitate these discussions. (3) Continuing educational (CE) programmes at member society meetings. Topics are prioritized based on input received from the local societies. Programmes often are those from CE courses given at meetings, such as conferences of the US Society of Toxicology (US SOT) and EUROTOX from the previous year. (4) Biennial Risk Assessment Summer School (RASS), an intensive week-long interaction between senior toxicologists who serve as faculty with attendees providing individual training. (5) Dissemination of donated printed toxicological books from publishers and syllabi from continuing education courses to regional locations. (6) Web-based interactive training programmes in regions where formal toxicological educational programmes are limited or lacking. (7) Preparation and distribution of monographs on selected topics of very current interest. Monographs on environmental oestrogens and genetically-modified foods have been published. The recent activities in each of these programmes are reviewed in this paper.

Education↗
PubMed2004

A multi-laboratory evaluation of a common in vitro pepsin digestion assay protocol used in assessing the safety of novel proteins.

Rationale. Evaluation of the potential allergenicity of proteins derived from genetically modified foods has involved a weight of evidence approach that incorporates an evaluation of protein digestibility in pepsin. Currently, there is no standardized protocol to assess the digestibility of proteins using simulated gastric fluid. Potential variations in assay parameters include: pH, pepsin purity, pepsin to target protein ratio, target protein purity, and method of detection. The objective was to assess the digestibility of a common set of proteins in nine independent laboratories to determine the reproducibility of the assay when performed using a common protocol. Methods. A single lot of each test protein and pepsin was obtained and distributed to each laboratory. The test proteins consisted of Ara h 2 (a peanut conglutin-like protein), beta-lactoglobulin, bovine serum albumin, concanavalin A, horseradish peroxidase, ovalbumin, ovomucoid, phosphinothricin acetyltransferase, ribulose diphosphate carboxylase, and soybean trypsin inhibitor. A ratio of 10U of pepsin activity/microg test protein was selected for all tests (3:1 pepsin to protein, w:w). Digestions were performed at pH 1.2 and 2.0, with sampling at 0.5, 2, 5, 10, 20, 30, and 60min. Protein digestibility was assessed from stained gels following SDS-PAGE of digestion samples and controls. Results. Results were relatively consistent across laboratories for the full-length proteins. The identification of proteolytic fragments was less consistent, being affected by different fixation and staining methods. Overall, assay pH did not influence the time to disappearance of the full-length protein or protein fragments, however, results across laboratories were more consistent at pH 1.2 (91% agreement) than pH 2.0 (77%). Conclusions. These data demonstrate that this common protocol for evaluating the in vitro digestibility of proteins is reproducible and yields consistent results when performed using the same proteins at different laboratories.

Clinical Laboratory Techniques↗
PubMed2003

The perceived healthiness of functional foods. A conjoint study of Danish, Finnish and American consumers' perception of functional foods.

Functional foods presumably enable the consumer to lead a healthier life without changing eating habits. Whether consumers accept this proposition or not is potentially influenced by their perceptions of the healthiness of the processing methods, enrichment components, food-types, and health claims used in the production and marketing of functional foods. Because consumers may perceive functional enrichment as interfering with nature, cultural values pertaining to man's manipulation of nature may also influence consumer acceptance of functional foods. The purpose of the study described here is to clarify to which extent Danish, Finnish and American consumers' perceptions of the healthiness of functional foods are explained by the factors mentioned above. The general results indicate that values pertaining to man's manipulation of nature is only modestly related to the acceptance of functional foods, whereas the use of different health claims, processing methods, enrichments, product types, and especially the interactions between the two latter, are important determinants of consumers' perceptions of the healthiness of functional foods.

Consumer Behavior↗
PubMed2004

Supervised identification of allergen-representative peptides for in silico detection of potentially allergenic proteins.

MOTIVATION: Identification of potentially allergenic proteins is needed for the safety assessment of genetically modified foods, certain pharmaceuticals and various other products on the consumer market. Current methods in bioinformatic allergology exploit common features among allergens for the detection of amino acid sequences of potentially allergenic proteins. Features for identification still unexplored include the motifs occurring commonly in allergens, but rarely in ordinary proteins. In this paper, we present an algorithm for the identification of such motifs with the purpose of biocomputational detection of amino acid sequences of potential allergens. RESULTS: Identification of allergen-representative peptides (ARPs) with low or no occurrence in proteins lacking allergenic properties is the essential component of our new method, designated DASARP (Detection based on Automated Selection of Allergen-Representative Peptide). This approach consistently outperforms the criterion based on identical peptide match for predicting allergenicity recommended by ILSI/IFBC and FAO/WHO and shows results comparable to the alignment-based criterion as outlined by FAO/WHO. AVAILABILITY: The detection software and the ARP set needed for the analysis of a query protein reported here are properties of the Swedish National Food Agency and are available upon request. The protein sequence sets used in this work are publicly available on http://www.slv.se/templatesSLV/SLV_Page____9343.asp. Allergenicity assessment for specific protein sequences of interest is also possible via ulfh@slv.se

Algorithms↗
PubMed2000

Have lessons been learned from the UK bovine spongiform encephalopathy (BSE) epidemic?

History is full of examples of public health, commerce and politics in conflict. In recent years attempts to protect UK egg producers, after the discovery of Salmonella enteritidis (phage type 4) in hens' eggs, strained previously good working relationships between medical and veterinary epidemiologists and ended the political career of a government minister who spoke out in defence of the public health. Against the background lessons of earlier high profile public health problems in the UK conflict should have been avoided when bovine spongiform encephalopathy (BSE) started. It might have been expected that its significance for human health could have been recognized and researched earlier. Public announcements about it could have been timely and clear. Unfortunately this was not the case and it looks as though similar mistakes are going to be repeated over genetically modified foods.

Animal Husbandry↗
PubMed2001

Research needs for human nutrition in the post-genome-sequencing era.

The sequencing and annotation of the human genome and the genomes of other model organisms offer new tools and new opportunities for human nutrition research in the 21st century. Basic research continues to be the key foundation for formulating solid nutrition recommendations for the public, but the basis for establishing human nutrient recommendations today suffers because of lack of good biomarkers and because of weak federal funding for nutrition research. In the context of this post-human genome-sequencing era, tantalizing opportunity exists in seven areas-four basic and three applied: 1) identification of molecular biomarkers for nutrient status; 2) characterization of single polynuclear polymorphisms (SNPs) associated with nutrition; 3) development of a national genome array nutrition database; 4) use of models at all phylogenetic levels for nutrition research; 5) application of post-genome-sequencing tools to study diet and human health, including phytonutrients and genetically modified foods; 6) application of these tools to the role of nutrition in pathogenesis of human disease; and 7) development of a funding base for research on exercise and human health.

Biomarkers↗
PubMed2003

Diet, nutrition, and cancer prevention: the postgenomic era.

The genomic era of human nutrition is upon us: the human genome and several plant genomes have been characterized, and genetically modified foods are now abundantly available in the marketplace. The link between diet and cancer is well established, and new genomic technologies have made possible the investigation of nutritional modulation of the carcinogenesis pathway with nutrients, micronutrients, and phytochemicals. Current study of nutrient-modulated carcinogenesis involves exploring the effect of nutrients on DNA damage and repair mechanisms; DNA methylation, which influences gene expression and cellular phenotypes; antioxidant rearranging and oxidative stress; target receptors and signal transduction pathways; cell cycle controls and check points; apoptosis; and antiangiogenic processes. With nutritional genomics, proteomics, and metabolomics, scientists are able to simultaneously elucidate the biological effects of dietary constituents on cell function and global gene expression. This generation of new knowledge on nutrient-gene interactions provides the justification for a research framework for diet and cancer prevention that is focused on identifying and developing new biomarkers as well as a novel and contemporary paradigm for dietary intervention.

Diet↗
PubMed2006

Computational detection of allergenic proteins attains a new level of accuracy with in silico variable-length peptide extraction and machine learning.

The placing of novel or new-in-the-context proteins on the market, appearing in genetically modified foods, certain bio-pharmaceuticals and some household products leads to human exposure to proteins that may elicit allergic responses. Accurate methods to detect allergens are therefore necessary to ensure consumer/patient safety. We demonstrate that it is possible to reach a new level of accuracy in computational detection of allergenic proteins by presenting a novel detector, Detection based on Filtered Length-adjusted Allergen Peptides (DFLAP). The DFLAP algorithm extracts variable length allergen sequence fragments and employs modern machine learning techniques in the form of a support vector machine. In particular, this new detector shows hitherto unmatched specificity when challenged to the Swiss-Prot repository without appreciable loss of sensitivity. DFLAP is also the first reported detector that successfully discriminates between allergens and non-allergens occurring in protein families known to hold both categories. Allergenicity assessment for specific protein sequences of interest using DFLAP is possible via ulfh@slv.se.

Algorithms↗
PubMed2005

A perspective on DNA microarray technology in food and nutritional science.

PURPOSE OF REVIEW: The functions of nutrients and other foods have been revealed at the level of gene regulation. The advent of DNA microarray technology has enabled us to analyze the body's response to these factors in a much more holistic manner than before. This review is intended to overview the present status of this DNA microarray technology, hoping to provide food and nutrition scientists, especially those who are planning to introduce this technology, with hints and suggestions. RECENT FINDINGS: The number of papers examining transcriptomics analysis in food and nutrition science has expanded over the last few years. The effects of some dietary conditions and administration of specific nutrients or food factors are studied in various animal models and cultured cells. The target food components range from macronutrients and micronutrients to other functional food factors. Such studies have already yielded fruitful results, which include discovery of novel functions of a food, uncovering hitherto unknown mechanisms of action, and analyses of food safety. SUMMARY: The potency of DNA microarray technology in food and nutrition science is broadly recognized. This technique will surely continue to provide researchers and the public with valuable information on the beneficial and adverse effects of food factors. It should also be acknowledged, however, that there remain problems such as standardization of the data and sharing of the results among researchers in this field.

Consumer Product Safety↗
PubMed2001

Limits of knowledge and the limited importance of trust.

Perceived risk and related attitudes have been implicated as major factors in many of the difficult policy problems that face modern society (nuclear power, genetically modified food, etc). Experts often argue that no or very small risks are involved; people are still worried. Why? The standard answer is lack of trust. Data on trust and risk perception, however, point to only a weak relationship between the two (r approximately 0.3). It is suggested here that the reason for the surprisingly minor importance of trust is that people believe that there are clear limits to how much science and experts know. Results are presented from studies of risk perception of the public, experts, and politicians. Politicians and members of the public believe that there are many unknown effects of technology and such beliefs were strongly related to their perceived risk. Experts on nuclear waste, on the other hand, seemed to believe that little is unknown in their field of expertise. Regression analyses of risk perception showed the unknown-effects factor to be a more important explanatory factor than trust for the public and politicians.

Journal Article↗
PubMed2004

The precautionary principle and/or risk assessment in World Trade Organization decisions: a possible role for risk perception.

Risk analysis has been recognized and validated in World Trade Organization (WTO) decision processes. In recent years the precautionary principle has been proposed as an additional or alternative approach to standard risk assessment. The precautionary principle has also been advocated by some who see it as part of postmodern democracy in which more power is given to the public on health and safety matters relative to the judgments of technocrats. A more cynical view is that the precautionary principle is particularly championed by the European Community as a means to erect trade barriers. The WTO ruling against the European Community's trade barrier against beef from hormone-treated cattle seemed to support the use of risk assessment and appeared to reject the argument that the precautionary principle was a legitimate basis for trade barriers. However, a more recent WTO decision on asbestos contains language suggesting that the precautionary principle, in the form of taking into account public perception, may be acceptable as a basis for a trade barrier. This decision, if followed in future WTO trade disputes, such as for genetically modified foods, raises many issues central to the field of risk analysis. It is too early to tell whether the precautionary principle will become accepted in WTO decisions, either as a supplement or a substitute for standard risk assessment. But it would undermine the value of the precautionary principle if this principle were misused to justify unwarranted trade barriers.

Animals↗