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The study was conducted to evaluate the effect of addition of L-cysteine to tris-citric acid (TCA) extender on the post-thaw quality of Sahiwal bull semen. For this purpose, two consecutive ejaculates were collected from three Sahiwal bulls using artificial vagina at weekly intervals for a period of three weeks (three replicates). Qualifying semen ejaculates were diluted (50×106 motile spermatozoa ml-1) in TCA extender having L-cysteine either 0.0 (control) or 0.5, 1.0 or 2.0 mM. Diluted semen was cooled to 4°C for 2 h, equilibrated for 4 h at 4°C, filled in straws at 4°C, kept in liquid nitrogen vapours for 10 min and then stored in the liquid nitrogen. Thawing was performed after 24 h of storage, at 37°C for 30 s. and the sperm motility, viability, plasma membrane and acrosomal integrity were assessed. Higher (P<0.05) sperm motility, viability, plasma membrane and acrosomal integrity were observed using extenders containing 1.0 or 2.0 mM compared to those containing 0.5 or 0.0 mM L-cysteine. It is concluded that addition of L-cysteine (to reach 1.0-2.0 mM) in TCA extender improves the post-thaw quality of Sahiwal bull semen.
The differences in properties of pyruvate kinase (EC 2.1.7.40) from normal tissues and animal or human tumors are described and their significance for various metabolic abnormalities is reviewed. The tumor variant gamma3 from M2 isoenzyme of pyruvate kinase sensitive to L-cysteine inhibition, when over-expressed, can be used as a marker of tumorigenic transformation. It seems that this variant represents a tumor-specific oncoprotein, involved in a novel metabolic strategy of energy generation during increased cell proliferation.
The present work was aimed to obtain information about age-dependent changes of γ-glutamyltransferase (GGT) activity and the levels of non-protein sulfhydryl com­pounds (NPSH) in rat kidneys. In addition, protein-bound cysteine (PB-Cys), sulfane sulfur compounds and reactive oxygen species (ROS) were estimated. The results indicate that the activity of GGT and NPSH levels in the kidneys are re­duced with age. At the same time, a significant increase in the level of protein-bound cysteine was observed. Simultaneously, the content of sulfane sulfur compounds was increased in the group of the oldest animals. These findings indicate that the capacity for extracellular glutathione degradation and, in consequence, the availability of cysteine for intracellular glutathione biosynthesis may be impaired. The increased PB-Cys level indicates potentiation of the thiolation reaction, i.e. development of pro­tein-mixed disulfides. These results reveal age dependent disturbances in the thiol-disulfide equilibrium in the kidneys which leads to an imbalance between pro- and antioxidatory processes.
Cytosolic fractions B (salted out between 51-70% ammonium sulphate saturation) from rat liver and Morris hepatoma 7777, containing pyruvate kinase (EC 2.7.1.40) M2 isoenzymes, were purified by affinity chromatography on Blue Sepharose CL-6B. When compared by polyacrylamide gel electrophoresis at pH 8.3, all three M2 pyruvate kinase variants from Morris hepatoma 7777 had lower mobilities (α2, β2, γ3) than the three corresponding variants (α1, β1, γ2) from normal rat liver. Using an automatic amino-acid analyser, significant differences in selected amino- acid content have been found in corresponding highly purified γ3 and γ2 variants from Morris hepatoma and normal rat liver, respectively. The γ3 -variant of the Morris hepatoma M2 isoenzyme had twice the amount of L-tyrosine and L-cysteine, and a content of L-serine higher by 20% than the corresponding γ2 variant of the normal rat liver M2 isoenzyme. It contained, however, significantly less dicarboxylic amino acids which explains its lower electrophoretic mobility. It showed also a decrease (by ca. 10%) in several other amino-acid content, corresponding to a 10% decrease in the tumour enzyme molecular mass.
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