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The aim of the present study was to verify the hypothesis that α-lactalbumin (LALBA) gene polymorphism may be a factor differentiating defence mechanisms in cattle. The association between LALBA polymorphism and the levels of blood indices characterising defence functions were examined in a group of 129 female calves and young heifers, produced by random mating of parental couples. The levels of blood diagnostic indices were determined four times: at 15 to 30 d, 3 months, 6 months, and 12 months of age. The genotypes distribution within the progeny was uneven (LALBA AA - 59.7%, LALBA AB - 35.7%, LALBA BB - 4.6%). At all times of the analysing, statistically significant differences between LALBA genotype groups were observed in the number of leukocytes (LALBA AA>LALBA BB), and the percentage of eosinophils (LALBA BB> LALBA AA). LALBA polymorphism also differentiated the percentages of lymphocytes (15 to 30 d, 3 months), neutrophils (15 to 30 d, 3 months, 12 months), basophils (6 and 12 months), and monocytes (6 months). Differences in the levels of total protein and γ-globulin were relatively low in animals aged 15 to 30 d, 3 months, and 6 months, but high in those aged 12 months (LALBA AB>LALBA AA>LALBA BB). There was a correlation between LALBA polymorphism and lysozyme activity (LALBA AA>LALBA AB> LALBA BB). Higher values of the NBT reduction tests were recorded in calves aged 15 to 30 d, as compared with older ones.
The aim of the study was to determine the relationship between the polymorphism of the ALA gene and the size of lymphocyte subpopulation expression of viral protein in cows naturally infected with bovine leukaemia virus. The studies were performed on a population of 97 Black-and-White breed cows, aged 3-8 years, from three large herds. The cows were examined three times in monthly intervals beginning from the second half of the first month after calving. Enzootic bovine leukosis diagnosis was based on the identification of p24 viral protein in the lymphocytes of the examined animals and on the white blood cell counts. The fluorescent antibody technique was used to detect p24 protein in the B (B-B2) and T CD4+ and CD8+ lymphocytes. The ALA gene polymorphism at the - 1689 position was assayed with the PCR-RFLP/Sdu I method. It was found that the ALA gene polymorphism differentiated the size of subpopulations of B-B2 lymphocytes and the T CD4+ lymphocytes expressing the p24 protein. This protein was more frequently registered in the B-B2 lymphocytes and in the T CD4+ and CD8 lymphocytes in cows with persistent lymphocytosis than in cows with the aleukaemic form.
A study was undertaken to analyse the effect of polymorphism of α-lactalbumin (ALA) gene at position -1689 on the susceptibility of lymphocytes to the infection with bovine leukaemia virus (BLV) and to undergoing apoptosis during the first three months after calving. The experiments covered 97 Black-and-White breed cows, aged 3-8 years, from three herds. The infection with BLV was diagnosed on the basis of the presence of the p24 protein of the virus identified by means of the fluorescent antibody test in peripheral blood lymphocytes. The BLV-positive and apoptotic cells were registered with the use of a fluorescence microscope. Analyses of white blood cell counts enabled separating the BLV- infected cows into groups with the aleukaemic form and with persistent lymphocytosis. The results obtained demonstrated a relationship between polymorphism of ALA gene and the susceptibility of lymphocytes to BLV infections and to undergoing apoptosis. The current study, together with the authors' previous research, suggests the possibility of ALA gene participation in mechanisms of BLV infection pathogenesis.
Limited proteolysis experiments can be successfully used to probe conformational features of proteins. In a number of studies it has been demonstrated that the sites of limited proteolysis along the polypeptide chain of a protein are characterized by enhanced backbone flexibility, implying that proteolytic probes can pinpoint the sites of local unfolding in a protein chain. Limited proteolysis was used to analyze the partly folded (molten globule) states of several proteins, such as apomyoglobin, a-lactalbumin, calcium-binding lysozymes, cytochrome c and human growth hor­mone. These proteins were induced to acquire the molten globule state under spe­cific solvent conditions, such as low pH. In general, the protein conformational fea­tures deduced from limited proteolysis experiments nicely correlate with those deriv­ing from other biophysical and spectroscopic techniques. Limited proteolysis is also most useful for isolating protein fragments that can fold autonomously and thus be­have as protein domains. Moreover, the technique can be used to identify and pre­pare protein fragments that are able to associate into a native-like and often func­tional protein complex. Overall, our results underscore the utility of the limited pro- teolysis approach for unravelling molecular features of proteins and appear to prompt its systematic use as a simple first step in the elucidation of structure-dynamics- function relationships of a novel and rare protein, especially if available in minute amounts.
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