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  • Biochemistry and Biotechnology  (1)
  • cardiac hypertrophy  (1)
  • electrophoresis under nondenaturing conditions  (1)
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  • 1
    ISSN: 1435-1803
    Keywords: cardiac hypertrophy ; labetalol ; catecholamines ; spontaneously hypertensive turkeys ; ventricularmyosin pattern
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary In most animal species, left ventricular hypertrophy due to pressure overload is associated with an advantageous increase of the “slow” V3 isomyosin. In contrast, in spontaneously hypertensive turkeys, the development of left ventricular hypertrophy is associated with the synthesis of a “fast” V1-like isomyosin, with high incidence of cardiac failure. This could be related to the high catecholamine levels found in these animals. This is why we studied the ventricular myosin pattern after lowering of blood pressure and regression of cardiac hypertrophy obtained by means of labetalol, an α- and β-blocking drug which inhibits the effects of catecholamines. From the 2nd to the 32nd week of age, 22 turkeys were treated with increasing doses of p.o. labetalol (from 20 to 35 mg/kg body weight daily) and 16 other turkeys were given daily p.o. placebo. Blood pressure and heart rate were periodically measured by an indirect method. After sacrifice, the degree of cardiac hypertrophy was evaluated by the biventricular weight to body weight ratio, ventricular myosin was purified, Ca++-activated ATPase activity assessed, and ventricular myosin pattern was determined by two-dimensional gel electrophoresis of myosin heavy chains. Plasma and cardiac catecholamines were measured by high performance liquid chromatography. Throughout the study period, blood pressure and heart rate were significantly reduced in the labetalol-treated animals as compared to the untreated ones. At the end of the study period, the ventricular mass was significantly lower in the labetalol group. Nevertheless, no differences were obscrved in ventricular myosin pattern and Ca++-activated ATPase activity levels between the two groups. In the labetalol group, an increase in plasma catecholamines and only a slight, but not significant, increase in cardiac catecholamines was found. These data indicate that in spontancously hypertensive turkeys, the synthesis of the “fast” V1-like isomyosin is not influenced by known pathophysiological stimuli like blood pressure, cardiac hypertrophy and catecholamines.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1435-1803
    Keywords: ventricular myosin ; pressure overload ; peptide mapping ; electrophoresis under nondenaturing conditions
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Chronic pressure overload induces a redistribution in myosin isoenzymes as demonstrated by Ca++-activated ATPase activity, electrophoresis under non-denaturing conditions and immunohistochemistry. We compared, in two groups of renal hypertensive rats and control rats, the isoenzymic patterns obtained by electrophoresis under non-denaturing conditions with those observed after heavy chains digestion with S. Aureus V8 protease. In the hypertensive animals in which a shift towards the “slow” V2 and V3 isomyosins was evident, peptide mapping always gave origin to a band which was not present in the controls. Since we consider this peptide as a marker of the redistribution towards the “slow” isoforms, peptide mapping according to Cleveland appears to be a simple and useful method to assess differences in isomyosin composition, at least between hypertrophic pressure-overloaded and normal rat ventricles. Moreover, in our experience this technique is simple, the patterns obtained from highly purified substrates are very reproducible and the digestion allows easy and clear comparisons.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 0263-6484
    Keywords: Myoblast ; proliferation ; integrin ; gene therapy ; antisense ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Myoblasts gene-engineered in vitro and then injected in vivo are safe, efficient options for gene therapy. While isolation of satellite cells is routinely achieved, their proliferation potential in vitro remains a limiting factor for cell transplantation under clinical conditions. We have studied the role of reversible inhibition of gene expression by antisense oligonucleotides on the proliferation of the myogenic cells. Addition of antisense oligonucleotides to myoblast cultures has been used to inhibit specifically the expression of the β1-integrin subunit gene. Here we show that the effects of multiple pulses of a phosphorothioate oligodeoxinucleotide antisense on the attachment to substrata and on the proliferation of myoblasts are dose-dependent. The addition of antisense to rat myoblasts caused rounding up of the cells and most of the cells became detached after several days in culture. A single pulse did not show any consistent effect, while in the presence of continously administered antisense, the relative numbers of myoblasts in the treated muscle culture increased. We have no evidence of inhibition of myoblast fusion under these conditions. On the other hand, [3H]-TdR incorporation, total DNA and total number of cells decreased in antisense-treated cultures thus demonstrating an inhibitory effect of the phosphorothioate oligonucleotides on DNA synthesis. These side-effects could be overcome by substituting the phosphorothioate by unmodified oligonucleotides, so decreasing the half-life of the antisense, but also its toxicity. The overall results suggest a potential role of integrin antisense strategy in modulating the potential of myoblasts to proliferate.
    Additional Material: 1 Ill.
    Type of Medium: Electronic Resource
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