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PubMedDate not supplied

Genetically modified and wild soybeans: an immunologic comparison.

Most traits introduced into genetically engineered crops result from the expression of new proteins. As the first step toward assessing the allergenic potential of genetically modified organism (GMO) food, immunologic and physicochemical characterizations are needed. We prepared crude extract from GMO soybeans, wild soybeans, curd, and soy milk and then performed sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). After acidification with HCl, the samples were separated to globulin and whey. To evaluate changes in protein composition, either the samples were heated or pepsin was added. Polymerase chain reaction with primer encoding the 35S-promotor and the 3-enol-pyruvyl-shikimat-5-phosphat-synthase gene were performed, respectively, to detect the GMO component. SDS-PAGE results showed definite protein bands at 80 kDa in GMO soybean, 50 kDa in wild soybean, and a similar distribution of protein bands was noticed below 40 kDa. It was difficult to observe protein distribution because of modifications that occurred during processing in soybean-processed products. After heating, proteins of GMO and wild soybeans showed similar distributions and no distinct bands were detected at 50 and 80 kDa. Although SDS-PAGE analyses of raw GMO and wild soybeans differed, the same protein bands of 68, 37, and 20 kDa were observed in the globulin fraction after acidification. After adding pepsin, 20- and 68-kDa bands were found preserved in GMO and wild soybeans. The polymerase chain reaction procedures with primers specific to GMO soybeans showed that GMO soybeans and some curd samples included a GMO component. The skin test results of 49 patients showed 13 positive results to wild soybeans and 8 positive results to GMO soybeans. One patient had a positive skin test result to GMO soybeans only. Sera from nine patients with positive skin tests to the crude extract and a positive capsulated allergen product test to the soybean antigen were used for the immunoblotting of GMO and wild soybeans. GMO soybeans revealed a unique strong immunoglobulin E binding band at 25 kDa in some patients and wild soybeans showed a strong immunoglobulin E binding band at 30-36 kDa. To assess the allergenicity of GMO food, more research, including a selection of controlled sample materials and immunoassays of qualified sera, is needed.

Child↗
PubMed2006

Genetic modifiers of the phenotype of mice deficient in mitochondrial superoxide dismutase.

Sod2-/- mice, which are deficient in the mitochondrial form of superoxide dismutase (MnSOD), have a short survival time that is strongly affected by genetic background. This suggests the existence of genetic modifiers that are capable of modulating the degree of mitochondrial oxidative damage caused by the MnSOD deficiency, thereby altering longevity. To identify these modifier(s), we generated recombinant congenic mice with quantitative trait loci (QTL) containing the putative genetic modifiers on the short-lived C57BL/6J genetic background. MnSOD deficient C57BL/6J mice with a QTL from the distal region of chromosome 13 from DBA/2J were able to survive for as long as those generated on the long-lived DBA/2J background. Within this region, the gene encoding nicotinamide nucleotide transhydrogenase (Nnt) was found to be defective in C57BL/6J mice, and no mature NNT protein could be detected. The forward reaction of NNT, a nuclear-encoded mitochondrial inner membrane protein, couples the generation of NADPH to proton transport and provides NADPH for the regeneration of two important antioxidant compounds, glutathione and thioredoxin, in the mitochondria. This action of NNT could explain its putative protective role in MnSOD-deficient mice.

Alleles↗
PubMed2002

Investigations on genetically modified maize (Bt-maize) in pig nutrition: fattening performance and slaughtering results.

A grower finisher performance trial with forty-eight pigs was designed to compare the growth performance of pigs fed diets containing either genetically modified (GM) Bt-maize (NX6262) or its parental maize (Prelude) line. During the experiment, the pigs were fed with a grower and a finisher diet both containing 70% maize investigated in a previously study which showed that they contained similar ME values and digestibility of crude nutrients. The pigs with an initial live weight of 23.9 +/- 3.0 kg were allotted to single boxes. During a 91 days growing period the pigs of both groups recorded equal performance in daily weight gain (AW) 815 +/- 93 vs. 804 +/- 64 g/d depending on equal amounts of feed intake 1.95 +/- 0.15 vs. 1.94 +/- 0.15 kg/d (parental vs. transgenic). The results confirm equal performance among growing-finishing pigs fed parental or genetically modified maize containing diets. For slaughtering the pigs were divided into 4 groups with a different duration of the finishing period. After slaughtering the carcass characteristic were registered.

Animal Feed↗
PubMed2001

Induction of potent human immunodeficiency virus type 1-specific T-cell-restricted immunity by genetically modified dendritic cells.

A novel technology combining replication- and integration-defective human immunodeficiency virus type 1 (HIV-1) vectors with genetically modified dendritic cells was developed in order to induce T-cell immunity. We introduced the vector into dendritic cells as a plasmid DNA using polyethylenimine as the gene delivery system, thereby circumventing the problem of obtaining viral vector expression in the absence of integration. Genetically modified dendritic cells (GMDC) presented viral epitopes efficiently, secreted interleukin 12, and primed both CD4(+) and CD8(+) HIV-specific T cells capable of producing gamma interferon and exerting potent HIV-1-specific cytotoxicity in vitro. In nonhuman primates, subcutaneously injected GMDC migrated into the draining lymph node at an unprecedentedly high rate and expressed the plasmid DNA. The animals presented a vigorous HIV-specific effector cytotoxic-T-lymphocyte (CTL) response as early as 3 weeks after a single immunization, which later developed into a memory CTL response. Interestingly, antibodies did not accompany these CTL responses, indicating that GMDC can induce a pure Th1 type of immune response. Successful induction of a broad and long-lasting HIV-specific cellular immunity is expected to control virus replication in infected individuals.

Animals↗
PubMed2003

[Studies on protein-based identification method of genetically modified capsicum].

The detection system based on protein is a method to evaluate the safety of genetically modified foods (GMF). Using cecropin BD gene in capsicum, a detecting method was set up. It is a system of evaluating the real expressive condition and safety of the foreign target protein of GMF. In this studies, with the preformative technic method, a satisfactory results by making use of hemolymph of immunized pupae of Antheraea pernyi as standard experimental material was achieved, comparing with the realities of the goal protein expressive condition of cecropin D gene in capsicum. The detecting steps were as following: the goal protein from material was extracted roughly, then with CM-Sepharose-FF ion-exchange chromatography twice, the goal protein was purified moderately. The purified product was identified by detecting the anti-bacterial activity, electrophoresis, biological auto-photography of the goal protein and MADDI-TOF mass spectrum. The results showed that the expressive foreign target protein in transgenic capsicum was in accordance with standard protein in the physical and chemical property, anti-bacterial activity and molecular weight. It indicated that expression of the target gene in capsicum is real, it corresponded to expected value. The separation, purification and identification methods of cecropin D were established in the study. By means of the comparative experiments about anti-bacterial activity and molecular weight of anti-bacterial peptide(ABP) from GM-capsicum and hemolymph of immunized pupae of Antheraea pernyi, the identification method of target protein from GM-capsicum was set up. The method is easy to be operated, fast and feasible.

Capsicum↗
PubMed1998

Improved adherence of genetically modified endothelial cells to small-diameter expanded polytetrafluoroethylene grafts in a canine model.

PURPOSE: A significant limitation to using genetically modified endothelial cells (ECs) to seed prosthetic grafts before implantation has been poor cell adherence to the graft lumen. Methodologic changes to improve cell adherence were evaluated in a canine carotid interposition graft model using 4 mm interior diameter expanded polytetrafluoroethylene. METHODS: ECs harvested from external jugular veins were grown in culture, with 80% of the cells from each culture transduced by incubation with an LXSN-type retroviral vector carrying a gene for human prourokinase and a neomycin resistance gene for selection in antibiotic G418. Control grafts had passive luminal coating with fibronectin and were seeded with transduced ECs immediately after G418 selection; these grafts were incubated for 2 days before implantation. Experimental grafts had fibronectin forcefully squeezed through the interstices and were seeded with ECs that had recovered in culture for 5 days after G418 selection; these grafts were incubated for 4 days before implantation. For each control (n = 9) and experimental (n = 12) graft, a graft prepared in the same fashion but seeded with the remaining autologous nontransduced cells was placed in the contralateral carotid artery. Grafts were explanted after 30 days and were evaluated for patency, thrombus-free surface area, and cell-free surface area. RESULTS: No significant differences in patency rates were seen between any groups. The thrombus-free surface area was improved for experimental grafts (90%) compared with control grafts (76%), but this improvement did not achieve statistical significance. The cell-free surface area for transduced cells on experimental grafts was 65% compared with 96% for control grafts (p = 0.021) and was comparable with that for nontransduced cells on both control grafts (62%) and experimental grafts (51%; p = 0.201). CONCLUSIONS: Adherence of genetically modified endothelial cells to small-diameter expanded polytetrafluoroethylene grafts in an in vivo physiologic flow model is significantly improved when cells have a more prolonged recovery from G418 selection, when the graft lumen is more uniformly coated with fibronectin before EC seeding, and when seeded grafts are left longer in culture before implantation to develop cell lining stability. The short-term patency rate of these seeded grafts is not affected by increased cell retention; long-term graft patency data and luminal healing require further evaluation.

Animals↗
PubMed2005

Evaluation of detection methods for genetically modified traits in genotypes resistant to European corn borer and herbicides.

Detection of genetically modified (GM) traits in corn (Zea mays L.) is urgently needed for preservation of genetic identity and marketing GM products. A laboratory study was conducted to evaluate the efficiency, accuracy, and reliability of different analytical methods to detect GM traits in corn. Samples with known fractions of GM concentrations (Bacillus thuringiensis [Bt], Liberty Link [LL] and stacked [Bt/LL] genes) from commercial seed companies and those derived from yellow kernels in a white corn ear, outcrossed by pollen of neighboring Bt hybrid, were tested by lateral flow quick test kits and by enzyme-linked immunosorbent assay (ELISA)-based test strips purchased from different companies. Liberty Link hybrids are resistant to glufosinate (Liberty or Basta) herbicides, whereas Bt corn is developed for the control of European corn borer (Ostrinia nubilalis). Selected samples of GM concentrations were also tested in a commercial laboratory using DNA-based polymerase chain reaction (PCR) test. The results demonstrated that qualitative lateral flow quick tests could detect samples containing 1% or higher Bt and 2% or higher LL concentrations within the recommended time limit of the test. Faint test lines for samples containing 0.5 to 1% Bt or 1 to 2% LL concentrations appeared if samples remained in the test cup overnight. ELISA test strips detected the Bt content semiquantitatively in the range of 0.5 to 2.0%. Grain samples derived from non-Bt corn outcrossed by neighboring Bt pollen had usually lower GM concentrations than commercial GM seed samples. Both ELISA- and DNA-based PCR tests distinguished samples with GM concentrations between 0.1 to 0.5%, but the precision of quantification at this range was very low and results were highly inconsistent.

Enzyme-Linked Immunosorbent Assay↗
PubMed2003

Assessment of screening methods for the identification of genetically modified potatoes in raw materials and finished products.

Qualitative polymerase chain reaction methods for the detection of genetically modified potatoes have been investigated that can be used for screening purposes and identification of insect-resistant and virus-resistant potatoes in food. The presence of the nos terminator from Agrobacterium tumefaciens and the antibiotic marker gene nptII (neomycin-phosphotransferase II) was demonstrated in three commercialized Bt-potato lines (Monsanto Co., St. Louis, MO, USA) and one noncommercial GM-potato product (high amylopectin starch, AVEBE, Veendam, The Netherlands) and allows for general screening in foods. For further identification, specific primers for the FMV promoter derived from the figwort mosaic virus, the CryIIIA gene (delta-endotoxin from Bacillus thuringiensis subsp. tenebrionis), potato leafroll virus replicase gene, and the potato virus Y coat protein gene, were designed. The methods described were successfully applied to processed potato raw materials (dehydrated potato powders and flakes), starch samples, and finished products.

Agrobacterium tumefaciens↗
PubMed2002

Genetically modified foods: hazard identification and risk assessment. Session summary.

During a Joint Society of Toxicologic Pathology (STP)/International Federation of Societies of Toxicologic Pathologists (IFSTP) International Symposium, held between June 24 and 28, 2001, in Orlando, FL, USA, there was a session entitled as "Genetically Modified Foods: Hazard Identification and Risk Assessment". The purpose of this session was to present and discuss the current situations in the US, European Union and Japan for the public concerns, safety assessments and regulations on genetically modified (GM) products used as foods or food ingredients. Assuming the wide and fast growing of the usage of GM products, it is the duty for us as toxicologic pathologists, to supply reliable data on their safety and possible risks or hazards as a world-wide basis to not only governments or regulatory agencies but also general public of our countries.

Animals↗
PubMed2001

[Usefulness of an immunoassay test TRAIT for detection of genetically modified Roundup ready soybean in food products].

The test based on immunoassay TRAIT Test for the specific detection of Roundup Ready Soybean was used for reference material in the form of dried powdered soy beans contained 0, 0.3, 1.25, 2.5% of genetically modified material, for soy beans declared as Roundup Ready and for soy products from Warsaw market. The detection limit was approximately 0.1% GMO on dry weight basis. Experiment was also carried out on heated soybeans. The positive results was obtained since temperature was under 65 degrees C during 15 minutes of heating grounded beans; above this temperature specific protein was not recognisable by the antibody. The TRAIT Test should be regarded as a qualitative method and could be recommended for screening purposes. Investigation demonstrated that above mentioned test was useful for detection of protein of genetically modified soybean in unprocessed products.

Enzyme-Linked Immunosorbent Assay↗
PubMed2003

Yield effects of genetically modified crops in developing countries.

Onfarm field trials carried out with Bacillus thuringiensis (Bt) cotton in different states of India show that the technology substantially reduces pest damage and increases yields. The yield gains are much higher than what has been reported for other countries where genetically modified crops were used mostly to replace and enhance chemical pest control. In many developing countries, small-scale farmers especially suffer big pest-related yield losses because of technical and economic constraints. Pest-resistant genetically modified crops can contribute to increased yields and agricultural growth in those situations, as the case of Bt cotton in India demonstrates.

Agriculture↗
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↗
PubMed2000

Analysis of genetically modified plant gene expression using GUS fluorimetry.

A fluorimetric assay method for the analysis of beta-glucuronidase (GUS) reporter gene expression in genetically modified plants is described. Optimization of this method for woody plants and a statistical approach suitable for comparisons of gene expression in different transformants or tissues of the same plant is described. Example data from elm (Ulmus procera) SR4 regenerant plants, shown to be genetically modified by PCR and DNA-DNA hybridizations, in which higher GUS expression levels are found in stems than in leaves demonstrates the utility of this approach.

DNA, Plant↗
PubMed2004

[Study on the teratogenicity effects of genetically modified rice with Xa21 on rats].

OBJECTIVE: To observe the effects of genetically modified rice with Xa21 on the development of rat embryos. METHODS: According to sex, weanling rats were divided into four groups: transgenic rice group, non-transgenic rice group, AIN93G negative control group and MATDA positive control group. The rats were fed with corresponding food for 90 days and mated. The development of maternal rats and embryos were observed. RESULTS: The body weight gain of pregnant rats and the body weight, body length and tail length of fetal rats in transgenic rice group were significant increased than those in positive control group. The number of death embryos and adsorption embryos, the malformation rate (appearance, viscera, skeleton) in transgenic rice group were lower than those in positive control group. There were no significant difference of all indicators among transgenic rice group, non-transgenic rice group and AIN93G negative control group. CONCLUSION: Compared with the non-transgenic rice, transgenic rice modified with Xa21 gene showed no significant differences in rat pregnant rate and embryo development.

Animals↗
PubMed2005

Identification of genetically modified potato (Solanum tuberosum) cultivars using event specific polymerase chain reaction.

Several genetically modified (GM) cultivars are registered in Canada although they are not currently in commercial production. The GM cultivars can be distinguished from the non-GM and other GM cultivars by analyzing the DNA nucleotide sequence at the insertion site of the transgene corresponding to a single transformation event in the plant genome. Techniques based on modified polymerase chain reaction (PCR) strategies were used to generate sequence information from the plant genome flanking the insertion site of transgenic DNA for specific GM potato events. The plant genome sequence adjacent to the transgenic insertion was used to design PCR primers, which could be used in combination with a primer annealing to one of the nearby inserted genetic elements to amplify an event specific DNA fragment. The event specific PCR fragments generated were sequenced to confirm the specificity of the method.

DNA, Plant↗
PubMed2002

Sensitive dependencies and separation distances for genetically modified herbicide-tolerant crops.

The amount of land available for the coexistent growing of both organic and genetically modified herbicide-tolerant (GMHT) crops depends on the separation distance between the two types of crop. The form of the decline in the proportion of land available for growing one of these crop types due to increasing separation distance is linear on a suitable scale, but with a slope and intercept that are sensitively dependent on the proportion of the other crop already present. Spatially explicit simulations from realistic scenarios indicate that a major increase in separation distances, currently under review by the UK government, may have serious implications for the future coexistence of organic and GMHT crops in the UK.

Computer Simulation↗
PubMed1999

Experimental microcosms as models of natural ecosystems for monitoring survival of genetically modified microorganism.

An experimental approach for investigation of genetically modified microorganisms (GMMO) introduced into model ecosystems to evaluate potential risk of propagation of recombinant plasmids in surrounding medium has been developed. The object of modeling was Escherichia coli Z905 strain with a recombinant plasmid with bacterial luminescence genes, which was introduced into water microcosms of different structure. The approach involves comprehensive investigation of GMMO at four hierarchical levels: molecular (retaining the structure of the plasmid and expression of cloned genes); cellular (variation of metabolic activity); population (competitive power and metabolic interactions of GMMO with indigenous microflora, migration of recombinant and natural plasmids); ecosystem (effect of GMMO and cloned genes on ecosystem parameters). The experimental evidence and theoretical estimates are intended to form grounds to develop a basic version of an ecological certificate for different GMMO variants.

Adaptation, Biological↗
PubMed2006

Novel surface display system for proteins on non-genetically modified gram-positive bacteria.

A novel display system is described that allows highly efficient immobilization of heterologous proteins on bacterial surfaces in applications for which the use of genetically modified bacteria is less desirable. This system is based on nonliving and non-genetically modified gram-positive bacterial cells, designated gram-positive enhancer matrix (GEM) particles, which are used as substrates to bind externally added heterologous proteins by means of a high-affinity binding domain. This binding domain, the protein anchor (PA), was derived from the Lactococcus lactis peptidoglycan hydrolase AcmA. GEM particles were typically prepared from the innocuous bacterium L. lactis, and various parameters for the optimal preparation of GEM particles and binding of PA fusion proteins were determined. The versatility and flexibility of the display and delivery technology were demonstrated by investigating enzyme immobilization and nasal vaccine applications.

Administration, Intranasal↗