Analytical importance of determination of lipase activity for the express analysis of contamination by heavy metals of sunflower seeds

During the last decades becoming more sharply there is a problem of chemical and environmental monitoring and industrial inspection the content of toxic elements in food raw materials and foodstuff. At the same time there is a need to develop rapid methods, informative, integral, reflecting not only the safety but also the ecological purity of food raw materials. The method of determination of content of toxic elements on activity of its own lipase of in situ (AОL-method) in seeds of oil-bearing crops, on the example of sunflower is offered. The system of mathematical assessment of analytical criteria of laboratory test used in clinical laboratory diagnostics was for this purpose adapted. Sunflower seeds in which established the maintenance of toxiferous elements served as an object of a research: Cd, Pb, As, Hg, by atomic absorption method on the KVANT-Z.ETA device. Further tests divided on clear, including high-quality and pollution-free, and polluted - naturally containing toxiferous elements and which are artificially contaminated. Definition of activity of a lipase of seeds was carried out by the standard titrimetric method. Decrease of the activity of enzyme was connected with the maintenance of toxiferous elements. Proceeding from the received results counted an analytical significance (sensitivity, specificity, and predictive value of descriptiveness the positive and negative results) of determination of level of maintenance of toxiferous elements in sunflower seeds by the AОL-method. The set values of analytical specificity of a method and predictive value of a positive take at the level of 77.3% and 71.4% respectively, do not allow to use the offered method for the quantitative analysis, however, analytical sensitivity at the level of 86.2% and the predictive value of the negative result at the level of 89.5%, allow to recommend the AOL-method for screening programs of chemical environmental monitoring and technological monitoring of a contamination of seeds of sunflower toxiferous elements

Gene polymorphisms of IL-1β (C511T), IL-17A (G197A), IL-12B (A1188C), TNFα (G308A) and IL-4 (C589T) associated with threat of early reproductive losses

Prevalence of threatened miscarriage is 16-25% of all pregnancies. The symptomatics of the threatened miscarriage in the first trimester may develop due to conversion of initial inflammatory reaction that disturbs the intersystemic and local interactions in the endometrium, followed by placental insufficiency, intrauterine fetal affection and spontaneous abortion. The aim of our work was to study the association between single nucleotide polymorphisms (SNPs) of some cytokine genes, e.g., IL-1β (C511T, rs16944), IL-17A (G197A, rs2275913), IL-12B (A1188C, rs3212227), TNFα (G308A, rs1800629), and IL-4 (C589T, rs2243250) and the risk of early reproductive losses among residents of Adyghe Republic (RA). The work was carried out at the Immunogenetic Laboratory of our Research Institute of Complex Problems. The allelic variants of cytokine genes were detected by SNP-method in 106 samples of genomic DNA in women with the threatened abortion in 1st trimester (n = 58) and the uncomplicated gestation (n = 48). SNP-typing of polymorphic variants of cytokine genes IL-1β, IL-17A, IL-12B, TNFα and IL-4 was carried out by PCR (polymerase chain reaction) with allele-specific primers and electrophoretic detection of results on test systems of (Litech, Moscow). Statistical analysis of experimental data was carried out by SPSS Statistical program 17.0. The correspondence of SNP distributions to expected values at Hardy—Weinberg equilibrium and comparison of allelic variants/ genotypes frequencies were performed using the χ2 criterion (Chi-square with Yates correction), odd ratios (OR) detected at significance level p < 0.05 and 95% confidence interval (95% CI). Reliability of the differences for the SNP frequencies for small samples was evaluated using the Fisher’s exact criterion. Heterozygous variant (C511T; OR = 3.46; 95% Cl: 1.04-11.54) and homozygous “mutant” genotypes (T511T; OR = 5.71; 95% Cl: 1.12-29.09) of the main proinflammatory IL-1β was significantly associated (p < 0.05) with the risk of developing threatening miscarriage in the Adygea residents. The -511Т allele of IL-1β gene, and -1188C variant of IL-12B gene increase the risk of the early termination of pregnancy, respectively, 5.8-fold (95% Cl: 2.4213.92; p = 0.00004), and 2.97-fold (95% Cl: 1.23-7.19; p = 0.01). The “mutant” -511T allelic variant of the IL-1 p gene is associated with the risk of developing a symptome complex of threatening miscarriage in Russian ethnic group (p = 0.0001; OR = 14.09), and in Adygea ethnic group (p = 0.02; OR = 8.17), which is almost undetectable in women with normal pregnancy in the first trimester. Thus, only С511Т (rs16944) in IL-1β gene and A1188C (rs3212227) in IL-12B gene of the five typed cytokine genes may be used as marker polymorphisms of gestational distress for the women in Adygea

Determination of heavy metals in sunflower seeds by atomic absorption spectroscopy based on fractional mineralization

The actual problem of food chemistry is the contamination of food raw materials and food products with heavy metals. Modern physico-chemical methods for the quantification of heavy metals are highly accurate and allow even trace amounts of elements to be determined, but the complexity of the sample preparation procedure is one of the most time-consuming steps, requiring high costs of reagents and time, especially for analysis of fat-containing samples. In this regard, development of new ways of determination of content of heavy metals in olive raw materials is relevant. Sunflower seeds in which they established the maintenance of toxic elements with use of different ways of sample preparation have acted as an object of a research: dry combustion and a wet mineralization and the developed new way of a fractional mineralization which distinctive features are: preliminary decomposition of test on fatty (oil) and fat-free (oilcake) of fraction with their independent mineralization. Oilcake is exposed to combustion at the maximum temperature 400 ?С for two full cycles, with convertion of the mineralizat in background solution and definition of metals by AAS-ETA method, oil – is dissolved in bipolar solvent in the ratio 1:6:2 from the mass of oil, with the subsequent direct input of mix in an analytical cell of the graphite tubular furnace of an electrothermal atomizer where there is a mineralization and simultaneous atomization of test. The use of fractional mineralization makes it possible to improve the metrological characteristics of the atomic absorption method of sunflower seeds: to increase the average openability of the investigated elements to 90.8%, to reduce the error of convergence (by 4.5%) and the reproducibility (by 4.9%) of the results of the study of cadmium and lead, and also to reduce the total time of sample preparation for 4–6 hours as compared with the test methods