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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Journal of molecular medicine 64 (1986), S. 786-792 
    ISSN: 1432-1440
    Keywords: Erythrocyte ; Heart muscle ; Receptor regulation ; (Na++K+)-ATPase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The assumption that the red blood cell can be used as a model for ouabain receptor regulation in heart muscle has been tested using isolated tissues from humans, guinea pigs, and chickens. The following results were obtained: 1. The affinity of the ouabain receptor was similar in both human erythrocytes and right atrial appendage, but the density of binding sites was much lower on the erythrocytes. There was no correlation between the binding capacity in both tissues. 2. Ouabain receptor occupation was closely correlated with inhibition of Na+/K+-transport in human erythrocytes and chick heart nonmuscle cells in culture. In contrast, in chick heart muscle cells, an occupation of 40% of the receptors decreased the Na+/K+-transport rate by only 10%. 3. In hypokalemia, the ouabain binding capacity was increased in human and guinea pig erythrocytes but not in guinea pig heart muscle. Such increases were seen in chick heart nonmuscle cells in moderate hypokalemia but in heart muscle cells only after severe hypokalemia. Incubation of chick heart muscle cells in toxic but not in “therapeutic” ouabain concentrations increased the number of ouabain receptors. Increases in receptor number attenuated the positive inotropic and toxic actions of ouabain. These variations between ouabain receptor regulation in red blood cells and heart muscle of several species may be attributable to the lack of a “sodium pump reserve” in erythrocytes and heart nonmuscle cells. Such variations indicate that the human erythrocyte is not a suitable model for the ouabain receptor in the human heart.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Journal of molecular medicine 63 (1985), S. 1253-1264 
    ISSN: 1432-1440
    Keywords: Cardiac glycosides ; Tolerance ; Heart cells
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary In cultured heart muscle cells from 10–13 day-old chicken embryos, the effects of acute (4 h) and chronic (3 days) exposure of the cells to varying concentrations of ouabain have been studied. In these cells, the cardiac glycoside ouabain binds to a specific cardiac glycoside receptor (KD=4 × 10−7 M; 750,000 receptors/cell). Binding to this receptor results in inhibition of active Na+/K+-transport [EC50 for active (86Rb+ + K+)-influx=4 × 10−6 M], and in an increase in beating velocity (“positive inotropic effect”;; EC50=4 × 10−7 M); toxic signs (arrhythmias) appear at concentrations ≥ 6 × 10−7 M. During exposure of the cells to 3 × 10−6 M ouabain for 3 days, tolerance develops with respect to both the positive inotropic and the toxic effect. The mechanism underlying this tolerance is identified as an increase in the number of active sodium pump molecules per cell, while the binding properties of the cardiac glycoside receptor remain unchanged. The development of cardiac glycoside tolerance is only observed in the presence of severe impairment of Na+/K+-homeostasis, due to cardiac glycoside-induced inhibition of active Na+/K+-transport. This, however, only occurs in the presence of toxic (receptor occupation ≥ 60%), but not in the presence of positive inotropic, non-toxic (receptor occupation 20–60%), ouabain concentrations. We conclude that the development of cardiac glycoside tolerance during long-term treatment in patients with heart failure should not occur with submaximal dose regimens, when toxic signs (arrhythmias) are absent.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Journal of molecular medicine 65 (1987), S. 308-316 
    ISSN: 1432-1440
    Keywords: Beta-blocker ; Withdrawal phenome-non ; Endogenous noradrenaline ; Heart cells
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary An in vitro model to evaluate the role of endogenous noradrenaline in the beta-blocker withdrawal phenomenon is described: Beating chicken heart muscle cells (5000 beta1-adrenoceptors/cell) and heart nonmuscle cells (3000 beta2-adrenoceptors/cell) were cultured in serum-free, hormone-supplemented medium. Basal state, subtype selective down-regulation of beta-adrenoceptors by endogenous noradrenaline (decrease in receptor number, beta1 more than beta2) was simulated by addition of noradrenaline to the culture medium; chronic beta-blockade was simulated by exposure of the cells for 3 days to various betablockers (propranolol, no ISA; timolol, slight ISA; pindolol, strong ISA). Beta-blocker withdrawal phenomenon — increased response in isoproterenol-induced cAMP production and positive inotropy — is correlated with the increase in the number of beta-adrenoceptors after withdrawal of the drugs. Propranolol induces a withdrawal phenomenon at every degree of noradrenaline-induced basal state down-regulation of beta-adrenoceptors; in contrast, a withdrawal phenomenon by pindolol is only seen at a higher degree of beta-adrenoceptor down-regulation. In the presence of physiological noradrenaline concentrations pindolol affects beta-adrenoceptor subtypes in a qualitatively different manner: the number of beta1-adrenoceptors is increased, the number of beta2-adrenoceptors is decreased. This finding demonstrates that the intrinsic sympathomimetic activity of nonselective beta-blockers can manifest itself only if the receptors are not strongly down-regulated. As beta2-adrenoceptors are present in a much less down-regulated state than beta1, ISA mainly acts on beta2-adrenoceptor subtype, thus, presenting a beta2-“pseudo-selectivity” of ISA.
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    Springer
    Journal of molecular medicine 62 (1984), S. 87-92 
    ISSN: 1432-1440
    Keywords: Ouabain binding ; Erythrocytes ; Diuretics ; Digitalis effect ; Receptor kinetics
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary It has been reported that during chronic treatment with digitalis, the number of digitalis binding sites is increased in human erythrocytes [22]. From this finding a tachyphylaxis for cardiac glycosides has been postulated. We reinvestigated this problem in several groups of patients. The number of3H-ouabain binding sites per erythrocyte in control persons (group I) was 214±60,n=43 (x±SD). The dissociation constant (KD) was 1.8±0.5 nM. Thirteen patients (group II) taking cardiac glycosides only, for at least 6 months, had 281±99 (p〈0.05) ouabain binding sites per single red cell, KD=1.8±0.7 nM. Group III (34 patients) took digitalis for more than 6 months and diuretics for at least 3 months (352±126 (p〈0.001), KD=1.6±0.6). Twenty-three of these (group IV) were taking a combination with “K+-saving” diuretics (336±194 (p〈0.01), KD=1.6±0.5) and (group V, 11 patients) a combination with “K+-losing” diuretics (462±133 (p〈0.001), KD=1.4±0.4). Nine patients (group VI) had a chronic hypokalemia, mainly due to taking furosemide (437±98 (p〈0.001), KD=1.5±0.4). Four control persons took 50 mg hydrochlorothiazide daily for more than 4 months without measurable K+-losses and without changes in ouabain binding sites. It is concluded from these findings that diuretic treatment with chronic hypokalemia in addition to digitalis is accompanied by a significant increase in ouabain binding sites in human red cells. Although the difference between control persons and those taking cardiac glycosides only, is statistically significant (p〈0.05), the biological relevance is questionable because of considerable overlap of the values. Receptor affinity was unchanged under all circumstances. A change in the number of ouabain binding sites — if occurring also in the heart — may go along with an altered digitalis sensitivity.
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1432-0738
    Keywords: Muscarinic acetylcholine receptor ; G-protein ; Oxime ; HGG-12 ; Cardiac tissue
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Direct interactions of the bispyridinium oxime HGG-12 with muscarinic acetylcholine receptors were investigated in porcine cardiac atrial membranes. Competition binding experiments using the radiolabeled muscarinic receptor antagonist (3H)QNB revealed specific binding of HGG-12 to muscarinic acetylcholine receptors of porcine atrial membranes with a dissociation constant of 3.8×10−7 mol/l. Muscarinic acetylcholine receptor-stimulated binding of the radiolabeled GTP analog (35S)GTP[S] to guanine nucleotide binding proteins (G-proteins) was used to study antagonistic and possible agonistic effects of HGG-12 at muscarinic acetylcholine receptors. HGG-12 completely inhibited carbachol- and oxotremorine-stimulated (35S)GTP[S] binding to pertussis toxin sensitive and insensitive G-proteins in a competitive manner. Inhibition constants (KI) of HGG-12 for blockade of carbachol- and oxotremorine-stimulated GTP[S]-binding (9.7×10−7 mol/l and 1.7×10−6 mol/l, respectively) were higher by about a factor of 100 than those of the muscarinic acetylcholine receptor antagonist atropine. In the absence of muscarinic acetylcholine receptor agonists, HGG-12 by itself had no stimulatory effect on (35S)GTP[S] binding in porcine atrial membranes. The results of this study show that the oxime HGG-12 is a competitive antagonist without intrinsic activity at porcine atrial muscarinic acetylcholine receptors. The stimulatory action of HGG-12 on muscarinic acetylcholine receptors which has been described by several authors is, therefore, suggested to be due to partial inhibition of acetylcholinesterase by the oxime rather than to direct agonism at muscarinic acetylcholine receptors.
    Type of Medium: Electronic Resource
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  • 6
    ISSN: 0005-2736
    Keywords: (Heart muscle cell) ; Cation flux ; Rb^+ uptake ; Vanadate
    Source: Elsevier Journal Backfiles on ScienceDirect 1907 - 2002
    Topics: Biology , Chemistry and Pharmacology , Medicine , Physics
    Type of Medium: Electronic Resource
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  • 7
    ISSN: 0005-2736
    Keywords: (Heart cell) ; Insulin ; K^+ uptake ; Na^+ uptake ; Ouabain ; Vanadate
    Source: Elsevier Journal Backfiles on ScienceDirect 1907 - 2002
    Topics: Biology , Chemistry and Pharmacology , Medicine , Physics
    Type of Medium: Electronic Resource
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  • 8
    ISSN: 0891-5849
    Source: Elsevier Journal Backfiles on ScienceDirect 1907 - 2002
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Type of Medium: Electronic Resource
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  • 9
    Electronic Resource
    Electronic Resource
    Amsterdam : Elsevier
    Free Radical Biology and Medicine 9 (1990), S. 158 
    ISSN: 0891-5849
    Source: Elsevier Journal Backfiles on ScienceDirect 1907 - 2002
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Type of Medium: Electronic Resource
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  • 10
    Electronic Resource
    Electronic Resource
    Amsterdam : Elsevier
    Biochimica et Biophysica Acta (BBA)/Bioenergetics 314 (1973), S. 224-241 
    ISSN: 0005-2728
    Source: Elsevier Journal Backfiles on ScienceDirect 1907 - 2002
    Topics: Biology , Chemistry and Pharmacology , Medicine , Physics
    Type of Medium: Electronic Resource
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