Radoslav Židek

Phytoestrogens dietary intake and health status of retiree from middle-notrh Slovakia region

Phytoestrogens found in foods of plant origin presents chemical substances that possess a wide range of biochemical benefits. It has been found that they contribute in different health related problems. A wide range of commonly consumed foods contain appreciable amounts of phytoestrogens. Consumption of diet rich to phytoestrogen acts as a protective factor against many diseases such as cardiovascular diseases, post-menopausal symptoms in the context of osteoporosis, cancerous illnesses of colon, prostate and breast. Three main classes of phytoestrogens covers: isoflavones, lignans and coumestans. Selected nine major phytoestrogens had been analyzed simultaneously in the same foods. Questionnaire designed to determine intake frequency as well as amount of selected foods and the most common diseases presented in the population has been used to find relationships between dietary habits and health status. Evaluation of selected goals in the present study has been realized in cooperation with 140 respondents in retired age (divided into Males - covered by 34 individuals and Females - 106 individuals), comming from middle-north Slovakia region. On the base of collected data it can be concluded, that evaluated population is presented by high values of lignans intake and particularly secoisolariciresinol, mainly caused by relative high proportion of cereals and linseed in the diet. Furthermore, the relationship between phytoestrogens intake and eating habits as well as its contribution in protection against selected diseases was demonstrated. Normal 0 21 false false false CS JA X-NONE

Comparison of the sensitivity of determining soyeabean allergens by ELISA method and SYBR green I

Normal 0 false false false SK JA X-NONE The aim of this study was to compare the suitability of two methods for detecting defatted soybean powder; SYBR Green I Real-time PCR and enzyme-linked immunosorbent assay (ELISA). Analysis of 20 artificially contaminated samples prepared by simple dilution with wheat flour revealed that both techniques were able to detect defatted soybean powder, although there were significant differences between the two methods. Wheat flour contamination with defatted soybean powder was detected in samples 1-5, (0.012 %; 120 mg.kg -1 ), but not in samples with lower contamination with soybean powder saples 6-20 using SYBR Green I real-time PCR. Samples 1-10 could not be quantified by ELISA as the absorbance values were greater than the detection limit, and while samples 11-20 were measured, only the values of samples 16, 17 and 18 were within the guaranteed quantification range specified by the ELISA kit manufacturer. Defatted soybean powder contamination was detected in samples 19 and 20, but absorbance values were highly similar to those of the negative control sample.

Comparison of microsatellite and blood group diversity among different genotypes of cattle

Genetic variability and relationships among five cattle breeds (Holstein, Pinzgau, Limousin, Slovak Spotted and Charolais) bred in the Slovak Republic were investigated separately using 11 microsatellite markers and 61 blood group systems. Allele frequency, heterozygosity (Ho, HE) and PIC values were investigated. F-statistics were computed separately. For microsatellite markers FIS, FIT, FST and for blood groups HS, HT, GST parameters were calculated. Microsatellite and blood group comparison showed similar results by F-statistics but some differences were marked using the other methods. Both methods were able to detect close relation between Slovak Pinzgau and Slovak Spotted cattle breeds. Their relation was confirmed by genetic distance, principal component analysis (PCA) and coefficient of admixture (mY). Important divergences between different markers used in the study were observed by the characterisation of Limousin and Charolais breeds.

Authentication of caprine milk and cheese by commercial qPCR assay

The objective of the study was to investigate potential adulteration of commercial caprine milks and cheeses with bovine milk using commercial qPCR assay. The assay comprised of bovine-, ovine- and caprine-specific primers and TaqMan probe and mammalian internal control. Specificity, sensitivity, linearity, reproducibility and efficiency of the bovine assay were tested as well. Specificity was verified by running reaction on the DNA of other milk-producing species (caprine and ovine) and made-up bovine-caprine (v/v) milk mixes. In both experiments, a bovine DNA fragment was amplified whereas no amplification was obtained from the other species. Sensitivity, linearity, reproducibility and efficiency were tested on 10-fold dilution series of 10 ng bovine DNA. The assay has shown good linearity (R 2 = 0.983) within whole range, with efficiency of 86% and excellent reproducibility (SD around the C T for the technical replicates <0.5). The sensitivity was adequate, as calculated LOD and LOQ were 1.44 pg and 2.94 pg of bovine DNA, respectively. Finally, the assay was used to authenticate 5 caprine milk samples and 5 caprine cheese samples, purchased from local supermarkets. Totally, 1 milk sample has shown the fluorescence signal, which exceeded baseline in cycle 39.01 ±0.69. However, the signal was above LOD and LOQ suggesting that there could not be unambiguously declared any adulteration with bovine milk. Amplification of bovine-specific DNA was not observed in the other samples indicating products were not adulterated. The commercial qPCR assay has proved that real-time PCR assays, as well as DNA-based techniques in a general, are the excellent and reliable tools for fighting with frauds in the food industry and protecting the public health. Normal 0 21 false false false SK X-NONE X-NONE

Detection of ovine milk adulteration using taqman real-time pcr assay

Food safety, quality and composition have become the subjects of increasing public concern. To prevent fraud and enhance quality assurance, credible analysis of dairy products is crucial. Bovine milk is more widely available and cheaper than milk of sheep and goat. Bovine milk is also processed in large quantities to produce a range of dairy produce. DNA-based methods have proven to be more reliable, because of the stability of DNA under the conditions of high temperature, high pressure, and chemical treatment used during the processing of some food products. The commercial InnuDETECT cheese assay based on the principle TaqMan real-time PCR systems have been tested for the identification and quantification of bovine DNA in ovine milk samples. DNA was extracted using the InnuPREP DNA Mini Kit and quantified by the QuantiFluor dsDNA system. The assay showed good linearity, with correlation coefficient of R2 = 0.983 and efficiency of 86%. The internal control amplified fragment from different mammalian species (cow, sheep and goat), with similar CT values. Detection of bovine DNA in milk mixtures was achieved even in samples containing 0.5% of bovine milk. The InnuDETECT cheese assay has been successfully used to measure bovine DNA in ovine milk, and will prove useful for bovine species identification and quantitative authentication of animal-derived products.

Assessing expression of TAS2R16 receptor on the tongue of elderly persons

In conducted study, we assessed expression of TAS2R16 receptor gene on the tongue of elderly persons. The TAS2R16 receptor belongs to family of G-protein coupled bitter taste receptors and is expressed in type 2 taste cells, which are a part of taste buds. The taste buds are distributed across the tongues surface on the specialised structures called papillae. The TAS2R16 receptor mediates bitter taste in response to β-glucopyranosides such as salicin. The purpose of conducted study was to examine, whether the ageing process influence gene expression and hence the perception of taste at the molecular level. Ageing process is often related to either decreased or total lost perception of taste qualities. It is due to physiological changes in the oral cavity. The changes in taste cell membranes involve altered function of ion channels and receptors, which ultimately lead to decreased tasting ability of elderly people. In addition, various causes, such as oral and systemic diseases, drug administration, lifestyle (i.e. smoking) and some oral conditions (wearing dentures, dental caries and coated tongue), may extracerbate this issue. Loss of taste may become a large factor in reduction of appetite, which may lead to malnutrition. To accomplish the objective of this study, we recruited ten elderly persons. One 25-year old human was used as calibrator. We used non-invasive scrapping method for collecting taste cells from fungiform papillae of each subject. A multiplex TaqMan real-time PCR was performed to amplify cDNA of TAS2R16 and PGK1 genes, whereas the last one served as housekeeping gene. The TAS2R16 gene expression for elderly persons relative to that of young one was calculated according to the 2 -Δ C T formula. Results pointed out to increased expression of TAS2R16 gene by 2-fold in 5 th and 8 th seniors. It is assumed that they perceive more intense bitterness from salicin at the molecular level than 25-year old person. The 2 nd , 3 rd , 7 th and 10 th elderly persons have had decreased expression level about 70%, whereas in case of 6 th one that was even about 90%. It is supposed that these subjects, in particular last one, respond to salicin very weakly. This data may show evidence of almost total loss of taste. The causes and consequences are discussed in more detail. Normal 0 21 false false false SK X-NONE X-NONE

Determination of the species specificity of the primers for the detection of chicken and turkey meat by realtime PCR method

The aim of this work was to use TaqMan Real-Time PCR for quantitative authentication of chicken and turkey meat. To meet this purpose, a specific pair of primers and TaqMan probe was used. The test was aimed at identifying the reaction cycle of turkey and chicken meat using by two sets of primers. With first set of primer designed for chicken we obtained the following results: Cp = 16.18 for 100% chicken DNA Cp = 29, 18 100% turkey DNA It was also amplified DNA of pig that exceeded the detection threshold fluorescence intensities in the 31.07 cycle (Cp = 31.07). Using primers designed for turkey we obtained the following results Cp = 31.16 for 100% CHDNA, Cp =16.18 100% TDNA. It was also amplified the 100% DNA of rabbit in 31.63 cycle (Cp = 31.63) and deer in cycle 32 (Cp = 32). The DNA of all other animal species was amplificated after more than 35 cycles (Cp >35). It follows that the second detection primer pair is specific enough to unrelated species of animals by 30 cycles of the reaction. Species authentication based on DNA analysis from this perspective overcomes all the shortcomings of proteins. At present, DNA analysis use different types of PCR. Is the most progressive Real-time PCR, which is suitable for the specific use of detection (primers and TaqMan probe). The TaqMan Real-time PCR is within the sensitivity and specificity, clearly one of the best methods for identifying the species of chicken and turkey meat. The specificity of this method, however, depends primarily on the specificity of the primers and TaqMan probe. The 30 cycle reaction was chosen by us as the threshold for specificity using primers for authentication chicken and turkey meat.

Analysis of gene expression in rabbit muscle.

Increasing consumer knowledge of the link between diet and health has raised the demand for high quality food. Meat and meat products may be considered as irreplaceable in human nutrition. Breeding livestock to higher content of lean meat and the use of modern hybrids entails problems with the quality of meat. Analysing of livestock genomes could get us a great deal of important information, which may significantly affect the improvement process. Domestic animals are invaluable resources for study of the molecular architecture of complex traits. Although the mapping of quantitative trait loci (QTL) responsible for economically important traits in domestic animals has achieved remarkable results in recent decades, not all of the genetic variation in the complex traits has been captured because of the low density of markers used in QTL mapping studies. The genome wide association study (GWAS), which utilizes high-density single-nucleotide polymorphism (SNP), provides a new way to tackle this issue. New technologies now allow producing microarrays containing thousands of hybridization probes on a single membrane or other solid support. We used microarray analysis to study gene expression in rabbit muscle during different developmental age stages. The outputs from GeneSpring GX sotware are presented in this work. After the evaluation of gene expression in rabbits, will be selected genes of interest in relation to meat quality parameters and will be further analyzed by the available methods of molecular biology and genetics.