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PubMed1998

Novel and transgenic food crops: overview of scientific versus public perception.

Recombinant DNA technology offers opportunities to develop new products in many different fields, including agriculture and the agro-food area. Transgenic plants with improved agronomic traits currently grow in field trials and a few varieties have already reached the European market. By and large, new technologies raise both concerns and expectations and modern biotechnology is no exception. Indeed, a voluntary moratorium on experiments involving recombinant DNA molecules was called for in 1974. At the present time, although a majority of academic and industrial scientists agree that transgenic food crops pose no risk for the environment or human health, some others believe that certain applications of modern plant biotechnology are hazardous. In particular, deliberate releases of genetically modified plants are regarded as risky. There is also a disparity between expert and lay perception of r-DNA technology applications to food crops, which makes public information a difficult task. This paper aims at exposing these conflicting points of view on the agricultural applications of modern biotechnology. We also propose some recommendations pertaining to public information in Europe. It appears that consensus conferences might be a good approach to stimulate public information and public debate in Europe, although this approach has to be adapted to the cultural context of each country.

Agriculture↗
PubMed2006

High-performance liquid chromatography and capillary electrophoresis for the analysis of maize proteins.

Methods for the analysis of maize proteins using HPLC and CE are reviewed. Most of the references cited in this review concern HPLC methods. Size-exclusion HPLC and especially RP-HPLC methods have been developed for characterization of normal and genetically modified maize, cultivar differentiation, and prediction of quality. Few CE methods for the analysis of maize proteins were found in the existing literature. Most of these methods focus on optimization of the separation of maize proteins using CZE and SDS-capillary gel electrophoresis.

Chromatography, High Pressure Liquid↗
PubMedDate not supplied

Management of herbicide-tolerant oilseed rape in Europe: a case study on minimizing vertical gene flow.

The potential commercialization of genetically modified herbicide-tolerant (GMHT) oilseed rape in Europe raises various concerns about their potential environmental and agronomic impacts, especially those associated with the escape of transgenes. Pollen of oilseed rape can be dispersed in space, resulting in the fertilization of sympatric compatible wild relatives (e.g. Brassica rapa) and oilseed rape cultivars grown nearby (GM and/or non-GM Brassica napus). The spatial and temporal dispersal of seeds of oilseed rape may lead to feral oilseed rape populations outside the cropped areas and oilseed rape volunteers in subsequent crops in the rotation. The incorporation of a HT trait(s) may increase the fitness of the recipient plants, making them more abundant and persistent, and may result in weeds that are difficult to control by the herbicide(s) to which they are tolerant. Vertical gene flow from transgenic oilseed rape to non-GM counterparts may also have an impact on farming and supply chain management, depending on labelling thresholds for the adventitious presence of GM material in non-GM products. Given the extent of pollen and seed dispersal in oilseed rape, it is obvious that the safe and sound integration of GMHT oilseed rape in Europe may require significant on-farm and off-farm management efforts. Crucial practical measures that can reduce vertical gene flow include (1) isolating seed production of Brassica napus, (2) the use of certified seed, (3) isolating fields of GM oilseed rape, (4) harvesting at the correct crop development stage with properly adjusted combine settings, (5) ensuring maximum germination of shed seeds after harvest, (6) controlling volunteers in subsequent crops, and (7) keeping on-farm records. The implementation of the recommended practices may, however, be difficult, entailing various challenges.

Agriculture↗
PubMed2003

Oryzacystatin I expressed in transgenic potato induces digestive compensation in an insect natural predator via its herbivorous prey feeding on the plant.

We observed recently that the rice cysteine proteinase inhibitor, oryzacystatin I (OCI) expressed in transgenic potato does not affect growth and development of the two-spotted stinkbug predator (Perillus bioculatus) via its herbivorous prey feeding on the plant. Here we monitored the inhibitory activity of recombinant OCI along this potato --> herbivore --> predator continuum, to determine if the absence of effect was associated with a digestive compensatory response of the predator following inhibition of its proteinases by the recombinant cystatin. After confirming that OCI is present in the plant, and ingested in an active form by potato beetle larvae, quantitative and electrophoretic assays allowed us to determine that the recombinant cystatin (representing about 0.8% of total soluble proteins in leaves) was entirely bound to a approximately 30-kDa target proteinase in the prey's midgut, forming a sodium dodecyl sulphate (SDS)-stable complex detected on immunoblots with an anti-OCI polyclonal antibody. Despite the apparent absence of free, residual OCI in the beetle's midgut, digestive protease activity in the predator, known to include OCI-sensitive activity, was altered negatively when the prey was fed the modified plant. This inhibitory process at the third trophic level was accompanied by a compensatory response in the predator, by which serine-type proteinases were synthesized de novo. Overall, our data suggest that the affinity between OCI and the predator's OCI-sensitive proteinases is: (i) as strong as (or stronger than) the affinity between OCI and the potato beetle 30-kDa-sensitive proteinase; and (ii) stronger than the affinity between these enzymes and the plant endogenous homologue of OCI, potato multicystatin, induced in the plant by potato beetle feeding. Our results also show that predatory organisms can adapt their digestive metabolism to the presence of plant antidigestive proteins ingested by their herbivorous preys. In a broader context, this study stresses the need to monitor the inhibitory effects of PI-expressing plants not only on the herbivorous insects targeted, but also on the organisms likely to consume these pests in the environment.

Animals↗
PubMed2003

Tracking genes from seed to supermarket: techniques and trends.

Analytical techniques to track plant genes in the environment and the food chain are essential for environmental risk assessment, government regulation and production and trade of genetically modified (GM) crops. Here, I review laboratory techniques to track plant genes during pre-commercialization research on gene flow and post-commercialization detection, identification and quantification of GM crops from seed to supermarket. At present, DNA- and protein-based assays support both activities but the demand for fast, inexpensive, sensitive methods is increasing. Part of the demand has been generated by stringent food labeling and traceability regulations for GM crops. The increase in GM crops, changes in GM crop design, evolution of government regulations and adoption of risk-assessment frameworks will continue to drive development of analytical techniques.

Enzyme-Linked Immunosorbent Assay↗
PubMedDate not supplied

Recombinant proteins in newly developed foods: identification of allergenic activity.

A number of agricultural crops are being modified for various purposes using recombinant DNA technology. Since transferred genes may code for proteins that are ordinarily not present, there is concern about the potential allergenicity of these new varieties. The safety evaluation of transgenic foods is relatively easy when the allergenicity of the gene source is known. Recombinant allergens in genetically engineered or altered foods can be identified using traditional immunological assays such as RAST or ELISA inhibition or immunoblotting procedures. Our recent studies of two corn proteins (10 kD and HSZ) used to alter grain amino acid composition and of transgenic soybeans with an altered fatty acid profile are examples of this approach. Both 10 kD and HSZ did not bind IgE antibodies from sera of corn-reactive subjects by immunoblotting. Studies of wild-type and transgenic soybeans with high oleic acidic content by RAST inhibition and immunoblotting with pooled sera of soy-allergic individuals demonstrated no difference in the allergen content of both extracts. In contrast to these studies, a recent investigation by Nordlee et al. (1996) of transgenic soybeans which expressed a methionine/cysteine-rich protein from Brazil nuts identified this protein as a major Brazil nut allergen. These studies indicate that, when the gene source is from a known allergen or if the recipient contains allergens, it is possible to determine whether the allergen content of the transgenic line is altered relative to the nontransgenic varieties.

Allergens↗
PubMed2005

The meaning of "natural": process more important than content.

The meaning of the desirable attribute "natural" was explored in two samples, American college students and adults in the Philadelphia jury pool. Participants rated the naturalness of a variety of "natural" entities, before and after they were transformed by operations such as freezing, adding or removing components, mixing with other natural or unnatural entities, domestication, and genetic engineering. Results support four hypotheses. First, the principle of contagion accounts for many aspects of the reduction of naturalness by contact with unnatural entities. Second, chemical transformations reduce naturalness much more than physical transformations do. Third, the history of an entity's processing is more important in determining its naturalness than is the nature of the entity's contents. Fourth, mixing like natural entities (e.g., water from different sources) does not markedly reduce naturalness. The insertion of a gene from another species, the process used in producing genetically modified organisms, produces the biggest drop in naturalness; domestication, a human-accomplished activity that changes genotype and phenotype in major ways, is considered much less damaging to naturalness.

Adult↗
PubMedDate not supplied

Controversy over genetically modified organisms: the governing laws and regulations.

Genetically Modified Organisms (GMOs) are increasingly becoming a topic of controversy in the U.S. and abroad. The public is questioning their safety and wanting the products labeled as genetically modified. There are other concerns from some of the scientific world and some government officials and organizations such as the Food & Agricultural Organization (FAO) that question whether adequate research has been done to qualify GMOs as safe for long-term use. Of particular concern are the allergenic properties, a GMO may impart, possible transfer effects of antibiotic resistance (given that antibiotic resistant marker genes are used for many GMOs), the expression of previously unexpressed traits, and the drift of pollen from genetically modified crops. It has also been noted that the laws and regulations governing the biotechnology world are outdated, are not comprehensive, and span too many agencies. The primary agencies currently regulating biotechnology are the U.S. Department of Agriculture (USDA), the Food and Drug Administration (FDA), and the Environmental Protection Agency (EPA).

Animals↗
PubMed2004

A transgenic dwarf rat model as a tool for the study of calorie restriction and aging.

We have previously reported a long-lived transgenic dwarf rat model, in which the growth hormone (GH)-insulin like growth factor (IGF)-1 axis was selectively suppressed by overexpression of antisense GH transgene. Rats heterozygous for the transgene (tg/-) manifest phenotypes similar to those in calorie-restricted (CR) rats. To further characterize the transgenic rat in comparison with CR rats, the present study evaluated glucose and insulin tolerance in tg/- and control Wistar (-/-) rats at 6-9 months of age. Rats were fed ad libitum (AL) or 30% CR from 6 weeks of age. In CR rats, glucose disposal after glucose load was facilitated without any significant surge of serum insulin, and insulin tolerance test also indicated increased insulin sensitivity. In transgenic rats, similar findings were observed after glucose and insulin load, and CR in tg/- rats further facilitated glucose disposal during glucose and insulin tolerance tests. These findings suggest the presence of both common and separate mechanisms regulating the glucose-insulin system between CR and the reduced GH-IGF-1 axis paradigms. The transgenic rat model is, therefore, a useful one for studies of CR and aging.

Aging↗
PubMed2005

Evaluation of an in vitro method for the measurement of specific IgE antibody responses: the rat basophilic leukemia (RBL) cell assay.

The evaluation of allergenic potential is a key parameter in the safety assessment of novel proteins, including those expressed in genetically modified crops and foodstuffs. The majority of allergic reactions to food proteins are immediate type hypersensitivity reactions in which the principal biological effector is IgE antibody; the accurate measurement of specific IgE antibody is therefore a critical factor in experimental systems designed to characterize protein allergenic potential. Due to the presence of much higher concentrations of other immunoglobulin isotypes, the assessment of specific serum IgE antibody poses substantial technical challenges. We have examined the utility of the rat basophilic leukemia (RBL) cell line for the measurement of murine IgE responses. RBL cells were sensitized with mouse monoclonal anti-dinitrophenyl (DNP) IgE antibody and challenged with DNP-albumin conjugates with various hapten substitution ratios (SR). Polyclonal anti-OVA IgE antisera were also assessed for activity in the RBL assay. Results were compared with titers measured in homologous passive cutaneous anaphylaxis (PCA) assay. Marked degranulation of RBL cells was induced by conjugates with SRs of between 16 and 32, whereas conjugates with lower SRs (of 10 or 3) failed to elicit significant serotonin release. All conjugates were able to induce mast cell degranulation in vivo in a PCA assay. Anti-OVA antisera with PCA titers of 1/32 to 1/64 failed to stimulate RBL cell degranulation, whereas high titer antibody (1/2048 to 1/4096 by PCA) induced a positive RBL cell response. Successful stimulation of RBL cell degranulation requires not only appropriate epitope densities but also high affinity antibody. These data indicate that this assay is inappropriate for the routine analysis of specific polyclonal IgE antibody responses such as those that are induced by exposure to complex protein allergens.

Allergens↗
PubMed2005

Relief for fish stocks: oceanic fatty acids in transgenic oilseeds.

Three recent reports (Baoxiu Qi et al., Amine Abbadi et al. and Anthony J. Kinney et al.) describe the production of very long-chain polyunsaturated fatty acids in transgenic plants. This might lead to a sustainable source of these valuable fatty acids for use in human food and animal feed. At present they are mainly available via consumption of fish, which is a limited and endangered resource.

Animal Feed↗