Authors:E. Takács, A. Nagy, É. Gelencsér, and A. Székács
Reliable determination of microbial or transgenic Cry toxins is an essential issue in food and feed analyses, and enzyme-linked immunosorbent assays (ELISAs) are the method of choice for quantifying these toxins currently in food and environmental analysis. Internal Quality Control (IQC) is an indispensable method to assess accuracy, precision, and reproducibility of analytical measurements. To assess the utility of the ELISA method, IQC was performed on EnviroLogix Cry1Ab/Cry1Ac QualiPlate ELISA with manufacturer supplied analytical standards. Applicability of negative and positive controls (C− and C+) was examined by Shewhart Control Charts for bias and Control Chart of the Range of Duplicates for precision. Linear regression (up to 5 ng ml−1 Cry1Ab concentration) of the commercial ELISA kit was compared to sigmoid calibration (up to 60 ng ml−1 Cry1Ab concentration). For immunoassay optimization process, possible matrix effects in different liquid and solid vertebrate tissues were examined by determination of the limit of detection values in these matrices.
Authors:S. Hussein, É. Gelencsér, M. Polgár, and György Hajós
Buffalo and cow milk caseins were submitted to hydrolysis either with á -chymotrypsin or with pepsin. Enzymatic peptide modification (EPM) was carried out by using L-methionine ethyl ester in the reaction mixture. As catalyst, á -chymotrypsin or pepsin was used. The incorporation of methionine in to the peptide chains in the presence of á -chymotrypsin showed an optimum value at 0.14 g Met added to the reaction mixture/1 g hydrolysate in both cases. In the case of pepsin used as catalyst, the optimal Met-enrichment was at 0.14 g Met added to the reaction mixture/1 g buffalo casein hydrolysate and at 0.34 g Met/1 g cow casein hydrolysate. The covalent nature of the amino acid incorporation was confirmed by SDS - polyacryl amide gel electrophoresis in the presence of urea. Electrophoretic patterns of the products indicate that transpeptidation plays an essential role in the EPM reaction. Antigenic character of the EPM- products was investigated in vitro by competitive indirect ELISA. Enzymatic peptide modification with methionine enrichment seems to be an efficient method for the reduction of the antigenic/potential allergenic character and for the improvement of the nutritive value of buffalo and cow milk caseins.
Authors:A. Nagy, Á. Baráth, J. Pauk, and É. Gelencsér
The authors have developed transgenic wheat lines with broad range of herbicide resistance. The transgenic wheat, containing bacterial derived alien gene (bar) regulated under the maize ubiquitin promoter, is resistant to glyphosinate (phosphinotrichin) agent family. The presence of bar gene expression product (phosphinotrichin acetyl transpherase enzyme, PAT) was confirmed by PAT-specific ELISA (Enzyme Linked Immuno Sorbent Assay). The qualitative and quantitative chemical composition of the transgenic wheat lines in comparison with their non-transgenic counterpart (year 2000-2002) and protein utilization of the wheat wholemeal flours (year 2002) were summarized. Nutritional evaluation of the protein was based on a rat model by using N-balance experiments. Among the protein sources, heat-treated samples were also introduced into the experimental diets. It was found that heat denaturation of the proteins led to results with somewhat increased biological value indices. The introduction of GM technology did not affect food intake or nutritional performance of the rats.
Authors:A. Nagy, J. Pauk, K. Takács, and É. Gelencsér
The expression levels of two marker proteins (phosphinotrichin acetyltransferase, PAT and wheat germ agglutinin, WGA) in the transgenic wheat lines and their resistance to digestion in small intestine of rats were studied in comparison with their non-transgenic counterpart obtained from green house and field experiments of two subsequent years. The marker proteins were quantified by ELISA. It was found that the expression of PAT and WGA markedly increased when the wheat was grown in the field compared to that in the greenhouse. There were no significant differences between the WGA contents of the parent and transgenic wheat lines, but a broad range of expression of PAT and WGA was observed in the transgenic lines. PAT did not survive in the small intestine of the rats, while WGA was resistant to digestion in different ratios and was bound to the intestinal epithelium.
Authors:A. Nagy, L. Jędrychowski, É. Gelencsér, B. Wróblewska, and A. Szymkiewicz
In recent years, research related to studying the effect of gut microflora on the human health has become of major economic importance. The main objective of our study was to examine whether or not the orally administered Lactobacillusstrains (LB) as an oral adjuvant can improve the mucosal immune protectionviaan enhanced IgA secretion to a co-administered marker antigen ovalbumin (OVA). We adapted a murine (BALB/c) model to demonstrate beneficial adjuvant effects of probiotic LB strains. Orally sensitised mice with OVA, which were prefed with native or heat denatured (HD) Lactobacillus salivarius (Ls) or Lactobacillus casei (Lc) responded better or with the same efficiency to a vaccination with antigen (OVA) than mice that had been sensitised only with OVA or not sensitised at all. Antibody (IgA) responses in the gut were increased in response to vaccination with OVA in mice that had been prefed with native or heat denatured Ls or Lc followed by Ls or Lc and OVA feeding. In prefed groups, the OVA feeding alone primed for specific immune response, while adjuvanted OVA has increased the immune exclusion potential of the gut.
Authors:E. Szabó, É. Gelencsér, E. Kovács, A. Jánosi, K. Takács, and E. Kiss
In our research we studied the occurrence of the main apple allergen coding gene-families (Mal d 1, Mal d 2, Mal d 3, Mal d 4) in 16 different and most preferably consumed apple varieties. After the DNA isolation by Wizard method the simple PCR reaction was used to examine the apple allergen-coding genes. To identify the presence of the four allergenic protein-coding genes two primer pairs were chosen. The presence of these allergens in most apple varieties could be confirmed. According to our results two varieties — Jonathan and Granny Smith — were found to contain the lowest amount of the coding genes of the allergenic apple proteins studied by us. Besides this, polymorph pattern was obtained by the use of Mal d 1 primer, which may lead to determine apple varieties with small amount of Mal d 1 allergens.The confirmation study of the presence of potential apple allergens by RNA and protein techniques is our plan in the near future.
Authors:K. Takács, E. Szerdahelyi, A. Nagy, and É. Gelencsér
Enzymatic hydrolysates of mechanically deboned meat (MDM) for a long time have been used as flavouring and functional food ingredients in the food industry and also as the bases of formula foods for special dietary uses.
The aim of the present study was to produce MDM hypo-antigenic products with improved digestibility and high biological value to be used as a milk protein alternative. turkey MDM was treated with digestive enzymes (trypsin and/or α-chymotrypsin, or pancreatin), followed by freeze drying. The optimised reaction conditions of hydrolysis were at 6% (w/v) of meat protein in 0.1% NaHCO3 buffer, pH 7.5; pancreatin enzyme with 50 TAME units/g meat protein substrate, 37 °C and 60 min). Hydrolysates (MDMH) were assessed for degree of hydrolyses (DH, %) by using trinitrobenzenesulphonic acid method and MW distribution by SDS-PAGE. Modification of immune reactive binding sites in MDMHs was monitored by immunoblot with cow’s milk, chicken egg or meat allergic human patients’ sera. Biological value indices (True Digestibility (TD), Net Protein Utilisation (NPU), Biological Value (BV)) were determined using rat feeding trials. Among the MDMH products, the pancreatic hydrolysate proved to be the most favourable in terms of biological value and digestibility as well as hypoallergenic property.
Authors:M. Hódsági, Á. Jámbor, E. Juhász, S. Gergely, T. Gelencsér, and A. Salgó
Resistant starches (RSs) are broadly investigated as appropriate additives in starch-based products due to their well-known and proved health benefits. However, it was shown in previous studies that these starches are sensitive of the different heat treatments used in the food processing, which can cause changes, especially in the resistance. There is an increasing trend to use microwave (MW) energy in food processing; therefore, our aim was to investigate the changes of RSs compared to native starches caused by MW heating. Maize, wheat, RS2 and RS4 starches were MW-treated according to a 2×2 experimental design (300 and 600 W of power, 30 and 150 s of time). The changes of in vitro digestibility, rheological properties (rapid visco analyser, RVA) and near infrared (NIR) spectroscopic characteristic were studied. Two spectrophotometers were applied (dispersive and Fourier-transform (FT)) to compare their sensitivity in the analysis of the MW-treated starches.Results showed that the digestibility of starches did not show any tendencies when increasing the microwave energy of treatments, the characteristics of the kinetic curves remained unchanged. The RVA analysis showed that the RSs did not gelatinize after the heat-treatments. The MW heating weakened the rheological properties of all starches. The NIR analysis was the most sensitive device for the detection of the effects of MW treatments. The analysis of the most characteristic carbohydrate regions (2080–2130 and 2270–2290 nm) highlighted structural alterations of the starches; moreover, the dispersive spectrophotometer was found to be more sensitive in the analysis of starches than the FT-one.
Authors:C. Cuadrado, É. Gelencsér, M. M. Perdosa, G. Ayet, M. Muzquiz, A. Puszati, György Hajós, and C. Burbano
The effect of germination conditions on the lectin of Lens culinaris var. Magda 20 seeds was studied. The seeds were germinated at 20 °C under different conditions of watering and light and for different periods of time. The seed lectin was assayed by haemagglutination and quantified by competitive ELISA. Changes in lectin content during germination were also monitored by SDS-PAGE and immunoblotting. Haemagglutinating activity and lectin content in the seeds were not changed during the first three days regardless of the conditions of the germination. However, lectin concentration was significantly higher after six days of germination; relative lectin levels being particularly high when germination was carried out in the light and with daily watering. The results of SDS-PAGE and immunoblotting have also shown that the lectin was not degraded during the first six days of germination however, other storage-proteins were broken down by proteolysis.