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101.
102.
The objective of this research were twofold: 1) to determine if decreased spermatozoal longevity, a previously reported heritable trait in chickens, was attributable to spermatozoal passage through the excurrent ducts, and 2) to document the morphology of the testicular excurrent ducts from affected roosters. Though spermatozoa were viable at ejaculation, as evidenced by their exclusion of ethidium bromide, fertility after intravaginal insemination of spermatozoa from affected roosters was less (p less than 0.001) than that observed with spermatozoa from nonaffected controls, 37 +/- 2.3 versus 58 +/- 1.5%, respectively, over a 21-day egg-collection interval. In contrast, fertility after intramagnal insemination of testicular spermatozoa from affected roosters was equivalent (p greater than 0.05) to that of nonaffected controls, 47 +/- 2.2 versus 41 +/- 3.6%, respectively. After intravaginal insemination, neither type of testicular spermatozoa fertilized oocytes. The ductuli efferentes proximales from affected roosters were characterized by a greater luminal cross-sectional area as well as a diminished height and number of longitudinal epithelial folds (p less than 0.005). It was concluded that heritable decreased spermatozoal longevity in the chicken is not attributable to an inherent spermatozoal defect. Rather, the defect is acquired during passage of spermatozoa through the extragonadal ducts of the rooster. 相似文献
103.
Non-covalent interactions result in aggregation of surface antigens of the parasitic nematode Trichinella spiralis. 总被引:4,自引:0,他引:4
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Surface antigens of three stages of the nematode worm Trichinella spiralis has been labelled with iodine and examined by sodium dodecyl sulphate (SDS)/polyacrylamide-gel electrophoresis under reducing and non-reducing conditions. A variety of interactions were defined: the infective larva surface antigens formed a spectrum of aggregates from 50kDa to greater than 1000kDa from subunits of 47kDa and 90kDa; in the adult worms of 60kDa complex arose fron interaction between two dissimilar molecules of 40kDa and 20kDa; the new-born larvae components formed homologous dimers from a 58kDa molecule. Aggregating molecules were adherent to lentil lectin-Sepharose and are therefore glycoproteins. The interactions observed were completely abolished by boiling in SDS/mercaptoethanol, but only partially destroyed by boiling in SDS/iodoacetamide. Based upon this, the associations can be characterized as non-covalent, but disulphide-bond-dependent. It is suggested, but not proved, that the aggregates arise from strong non-covalent hydrophobic interaction sites which are stabilized by intrachain disulphide bonds in the molecules concerned. 相似文献
104.
Increased Salsolinol Levels in Rat Striatum and Limbic Forebrain Following Chronic Ethanol Treatment 总被引:1,自引:1,他引:0
Abstract: Endogenous levels of salsolinol and its methylated metabolite were measured by combined gas chromatography and mass spectrometry in rats chronically exposed to ethanol for 150 days. The chronic ethanol administration produced a significant increase of salsolinol concentrations in dopamine-rich brain areas, e.g., the striatum and the limbic forebrain. A negative correlation was observed between plasma ethanol concentration and the level of salsolinol in the brain. A possible role for salsolinol in the regulation of ethanol drinking and/or in the development of ethanol dependence is discussed. 相似文献
105.
Demonstration of 2',3'-Cyclic Nucleotide 3'-Phosphohydrolase in Cultured Human Schwann Cells 总被引:6,自引:5,他引:1
Abstract: Schwann cell cultures were established from adult human sural nerve biopsies. 2'3'-Cyclic nucleotide 3'-phosphohydrolase (CNPase) activity was estimated in the homogenates of those cells by a sensitive isotope assay using [3 H]2',3'-cyclic AMP as substrate. A high level of CNPase activity was observed in cultured Schwann cells, whereas cultured human muscle and skin fibroblasts contained negligible levels of CNPase activity. CNPase of human Schwann cells followed typical enzyme-substrate kinetics, with an apparent K m of 1.6 m M for 2',3'-cyclic AMP, and the enzyme was stimulated by detergents such as Triton X-100 and deoxycholate. It was inhibited by p -chloromercuricbenzoate and 2'-AMP. These properties are typical of CNPase isolated from adult brain and spinal cord. CNPase can serve as a new biochemical marker of normal cultured human Schwann cells and can be useful in analyzing the properties of cultured Schwann cells from patients with dysschwannian neuropathies. 相似文献
106.
The beta-adrenergic receptor in human lymphocytes: subclassification by the use of a new radio-ligand, (+/-)-125 Iodocyanopindolol 总被引:5,自引:0,他引:5
(±)?125Iodocyanopindolol (ICYP), a new radio-ligand with high affinity and specificity to β-adrenoceptors was used to identify and characterized β-adrenergic receptors in human lymphocytes. Binding of ICYP was saturable with 1.56 ± 0.2 fmol ICYP specifically bound/106 cells at maximal occupancy of the sites and of high affinity (KD=57 ± 7.1pM, N=4. In contrast to 125Iodohydroxybenzylpindolol ICYP-binding was not affected by phentolamine (up to 10?4M) or serotin (up to 10?5M). Analysis of inhibition of ICYP-binding via a pseudo-Scatchard-plot (“Hofstee-plot”) by β1-selective (practocol, metaprolol) and β2-selective (IPS 339, zinterol) adrenergic drugs resulted in linear plots suggesting the existence of a homogeneous population of β-adrenergic receptorsin human lymphocytes. From the resulting KD-values for practolol (16.8 μM), metoprolol (4.11 μM), zinterol (0.08 μM) and IPS 339 (0.002 μM) is concluded that the β-adrenergic receptor present in human lymphocytes is of the β2-subtype. According to its low non-specific binding and its high specificity to β-adrenergic receptors ICYP appears to be an ideal ligand for long-term studies on the regulation of β-adrenergic receptors of human lymphocytes. 相似文献
107.
Summary H-Y antigen was determined in eight transsexual patients. Two of the four male-to-female transsexual patients typed as H-Y antigen-negative, while the other two typed as expected from their phenotypic and gonadal sex, namely H-Y antigen-positive. Of the four female-to-male transsexual patients, three typed as H-Y antigen-positive and one was H-Y antigen-negative, as expected. The presence of normal testes in H-Y antigen-negative males is assumed to result from a mutation of nucleotide sequences of the H-Y structural gene for antigenic determinants. Thus, an H-Y is produced with normal receptor-binding activity which can sustain the testis determination of the bipotent gonadal anlage. In the case of H-Y antigen-positive females with normal ovaries a deletion of the autosomally located H-Y structural gene is assumed. This deletion should affect sequences for repressor-binding (as was suggested for H-Y antigen-positive XX-males) and for receptor-binding activity of the H-Y antigen molecule. The resulting H-Y antigen is unable to bind to the gonadal receptor of the bipotent gonadal anlage. Thus an ovary is determined. The relevance of H-Y antigen for the aetiology of transsexualism is discussed. 相似文献
108.
U Quast J Engel E Steffen G Mair H Tschesche H Jering 《European journal of biochemistry》1975,52(3):505-510
Equilibrium measurements of the binding of reactive-site-cleaved (modified) bovine trypsin-kallikrein inhibitor (Kunitz) to alpha-chymotrypsin and beta-trypsin show a stoichiometric 1:1 association with high binding constants. At least in the case of chymotrypsin much evidence is presented that the reaction with modified inhibitor leads to the same complex as the reaction with virgin inhibitor does. The association rate constant of modified inhibitor with chymotrypsin at pH 7, 22.5 degrees C is 15.8 M-1 S-1. This is about 2 x 10(4) times slower than the binding of virgin inhibitor to that enzyme. In the analogous reaction of modified inhibitor with beta-trypsin, however, the association rate constant (1.2 x 10(4) M-1 s-1 at pH 6.9, 22.5 degrees C) is of about the same order of magnitude as it is in the reaction of virgin inhibitor and trypsin. These and analogous phenomena observed in the reactions of virgin and modified soybean trypsin inhibitor (Kunitz) with alpha-chymotrypsin and beta-trypsin suggest that the specificity of both inhibitors to trypsin is strongly reflected in the association rate constants of the modified forms. The dissociation rate constants of the complexes of trypsin-kallikrein inhibitor with chymotrypsin or with trypsin towards the modified inhibitor are estimated to be unmeasurably slow (half-life times of 45 or 1.5 x 10(4) years, respectively). 相似文献
109.
The association of alpha-chymotrypsin with basic pancreatic trypsin inhibitor was studied using extrinsic signals produced by fluorescent and nonfluorescent labels. The reactive dyes were covalently bound to the proteins in the complexed state, in which the binding region was protected. The signals were sufficiently large to measure the complex formation at protein concentrations of 10(-9)M by fluorescence and down to 10(-6)M by absorption. Therefore, the association and dissociation could be followed over a broad range of concentration. Good correspondence was observed between data which were obtained with different labels and with published values for the unlabeled proteins. Existing differences could be explained by different buffer conditions used by the different authors. Also the pH dependence of the dissociation rate constants was essentially unaltered by the introduction of the labels. The large signals allowed a direct measurement of the equilibrium constants of dissociation, even at high pH, at which they are in the range of 10(-8)M. The experimentally determined binding constants were in agreement with those calculated from the rate constants. The temperature dependence of the binding constants revealed a small positive and pH-dependent enthalpy change [deltaHo = 4.0 kcal/mol (16.7 kJ) at H 7.0[. The results prove that the labeling can be performed in such a way that the equilibrium and kinetic parameters of the system studied are not significantly influenced. 相似文献
110.
The time-course of inactivation of bovine liver glutamate dehydrogenase by pyridoxal 5'-phosphate was studied in the presence of varied amounts of 2-oxoglutarate or NADH. Pseudo-first-order analysis reveals that the protection by both these compounds is competitive with respect to the chemical modifier. The competition is only partial, however: saturation with either NADH or 2-oxoglutarate decreases the rate constant for inactivation to a finite minimum and not to zero. Similarly, the plot of activity at equilibrium as a function of the concentration of the protecting substrate or coenzyme reveals that neither NADH nor 2-oxoglutarate protects completely against inactivation. In initial-rate experiments, pyridoxal 5'-phosphate, used as an instantaneous inhibitor rather than a long-term inactivator, displayed non-competitive inhibition with respect to both 2-oxoglutarate and NADH. These results clearly indicate that, although there is mutual hindrance between the binding to the enzyme of pyridoxal 5'-phosphate, on the one hand, and 2-oxoglutarate or NADH on the other, binding is not mutually exclusive. These findings are discussed in terms of the two-step mechanism for inactivation by pyridoxal 5'-phosphate. It is concluded that lysine-126 cannot be solely responsible for binding either the substrate or the coenzyme, but could be essential for the catalytic step. 相似文献