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Biomedical subjects

N S Dhalla

Publications and source records attributed to N S Dhalla.

At least 235 records · Page 13Linked to original sources
PubMed1983

Stimulation of calcium pump activity in heart sarcolemma by timolol.

The effects of beta-adrenergic blocking agents, timolol and atenolol (1-1000 microM), were studied on rat heart sarcolemmal ATPase and Ca2+ binding activities. Timolol, unlike atenolol, increased both Ca2+-stimulated ATPase and ATP-dependent Ca2+ binding; the maximal effects were seen at 1 microM concentration of timolol. Both timolol and atenolol did not alter the sarcolemmal Mg2+ ATPase and nonspecific Ca2+ binding activities. Sarcolemmal Ca2+-stimulated ATPase was also activated by concanavalin A (6-66 micrograms/mL) which is known to alter membrane fluidity; however, Mg2+ ATPase was unaffected by this agent. These results indicate that timolol may stimulate Ca2+ pump activity in heart sarcolemma by changing membrane fluidity in a manner similar to that of concanavalin A.

Animals↗
PubMed1983

Physiological and pathophysiological aspects of cardiac prostaglandins.

It has now been demonstrated that prostaglandins produce a wide variety of actions on cardiac tissue. These effects are quite complex and are dependent on such factors as type of prostaglandin, animal species, experimental format, as well as the amount of prostaglandin employed. The last factor is of particular concern since the administration of large amounts of prostaglandins may not truly reflect the actual actions of the endogenously synthesized prostaglandins which are likely to be at a very low concentration. In this review we have attempted to analyze the pharmacological properties of prostaglandins on cardiac tissue including coronary arteries and myocardium as well as the subcellular basis for their actions. The possible reasons for the diverse effects of prostaglandins under different experimental conditions have been pointed out. An effort has also been made to synthesize the existing information concerning the actions of prostaglandins on heart in terms of current concepts. A discussion is presented concerning the role of endogenously synthesized prostaglandins in physiological and pathological aspects of cardiac performance and the potential use of prostaglandins and prostaglandin synthesis inhibitors in the treatment of heart disease.

Animals↗
PubMed1983

Sarcolemmal Na+-K+-ATPase activity in diabetic rat heart.

Heart sarcolemmal membranes were isolated by the hypotonic shock-LiBr treatment from rats with chronic diabetes induced by a streptozotocin (65 mg/kg, iv) injection. Sarcolemmal Mg2+-dependent ATPase activity was elevated, whereas 5'-nucleotidase and K+-p-nitrophenylphosphatase activities in diabetic heart were depressed in comparison to control preparations. Although patent Na+-K+-ATPase and patent ouabain-sensitive Na+-K+-ATPase activities were unaltered, latent Na+-K+-ATPase activities, as determined in membranes after alamethicin or deoxycholate treatments, were found to be significantly depressed in diabetic animals. A depression in the latent Na+-K+-ATPase activity in diabetic preparations was also observed in membranes prepared by the sucrose density gradient method. Insulin-treated diabetic rats were observed to have normalized latent Na+-K+-ATPase activities. Total phospholipid content did not differ, but cholesterol content of the sarcolemmal membranes was significantly increased in diabetic heart preparations. Sarcolemmal Na+-K+-ATPase activity in diabetic heart was more resistant to treatments with filipin, an agent known to bind with cholesterol residues. These results suggest that chronic experimental diabetes is associated with some defects in sarcolemmal enzymatic activities and composition.

Adenosine Triphosphatases↗
PubMed1983

Defective sarcoplasmic reticular calcium transport in diabetic cardiomyopathy.

The effects of insulin and thyroid hormone treatments on cardiac sarcoplasmic reticular function were investigated in chronic streptozotocin-induced diabetes in rats. ATP-dependent Ca2+ transport and Ca2+-stimulated ATPase activities were depressed significantly in microsomal samples from diabetic rats in comparison with control (P less than 0.05). This defect was seen at various times of incubation (1-20 min) and different concentrations of free Ca2+ (10(-7) to 10(-5) M Ca2+) and was accompanied by changes in the protein composition and phospholipid contents of the microsomal fraction. The defect in calcium transport in microsomal vesicles was not evident until 28 days after streptozotocin (65 mg/kg iv) injection, whereas increases in plasma glucose levels due to insulin-deficiency occurred within 3 days. All changes in function and composition of the sarcoplasmic reticulum were reversed by insulin administration to the diabetic rats. Although the plasma level of thyroid hormone was decreased in the diabetic rat, thyroid hormone treatment did not restore microsomal calcium transport in the diabetic animals. The results of this study provide some evidence that the depression in cardiac sarcoplasmic reticular calcium accumulation during diabetes is a consequence of insulin deficiency and associated chronic metabolic changes but the hypothyroid condition that accompanies experimental diabetes does not appear to play any role in this defect.

Adenosine Triphosphatases↗
PubMed1983

Characterization of Ca2+ release from the cardiac sarcoplasmic reticulum.

The characteristics of Ca2+ release in relation to Ca2+ binding were studied in sarcoplasmic reticulum vesicles isolated from canine myocardium. The Ca2+ binding appeared to be dependent on ATP as a 4 fold increase in Ca2+ binding was observed upon the addition of ATP. In the presence of a suboptimal ATP concentration (20 mumol/l; without ATP regenerating system) a rapid release of Ca2+ started within 2 min. The rate of Ca2+ release was increased by increasing the concentration of Ca2+ in the preincubation medium when studied by diluting preloaded vesicles in medium free of Ca2+ and ATP; an apparent saturation was reached at 5 mmol/l Ca2+ but Ca2+ release again increased between 5 and 10 mmol/l Ca2+. High pH (8.0) enhanced the Ca2+ release process. When Ca2+ loaded vesicles were treated with various phospholipases and proteases, an enhanced Ca2+ release was observed in comparison to the control values. The release of Ca2+ was also increased by pharmacological agents like caffeine, ether and halothane. The Ca2+ release rate was stimulated by the p-chloromercurybenzoate treatment, which decreased ATP dependent Ca2+ binding and Ca2+-stimulated ATPase activities of the sarcoplasmic reticulum vesicles. The effect of temperature when evaluated by Arrhenius plots showed a higher energy of activation of Ca2+ release (66.15 kJ/mol) in comparison to that for Ca2+ binding (41.03 kJ/mol). These results indicate that, although Ca2+ release and Ca2+ binding activities of the cardiac sarcoplasmic reticulum appears to be related, Ca2+ release is probably a distinct process and is controlled differently. It seems that the Ca2+ release site in sarcoplasmic reticulum membranes is lipoprotein in nature.

Adenosine Triphosphatases↗
PubMed1982

Purification and characterization of a Ca2+-dependent ATPase from rat heart sarcolemma.

The Ca2+-dependent ATPase was solubilized from rat heart sarcolemmal membranes upon digestion with trypsin and was found to be different from Ca2+-stimulated Mg2+-dependent ATPase (Dhalla, N. S., Anand-Srivastava, M. B., Tuana, B. S., and Khandelwal, R. L. (1981) J. Mol. Cell. Cardiol. 13, 413-423). The enzyme was purified by high speed centrifugation, ammonium sulfate fractionation, and column chromatography and was seen as a single protein band in nondenaturing polyacrylamide gel electrophoresis. In sodium dodecyl sulfate-acrylamide gels, the enzyme dissociated into two subunits or fragments with molecular weights of about 55,000 and 12,000. The molecular weight of the enzyme, estimated by gel filtration on a Sephadex G-100 column, was found to be about 67,000. The enzyme utilized ATP with a Km of 0.20-0.26 mM but was also able to utilize ITP, CTP, GTP, and ADP as substrates at much lower rates. It was activated by Ca2+ with a Ka of 0.13-0.21 mM; it was also activated by other cations in the order Ca2+ greater than Mn2+ greater than Sr2+ greater than Ba2+ greater than Mg2+. Divalent cations like Cu2+, Ni2+, and Mg2+ were potent inhibitors. The enzyme was insensitive to ouabain, verapamil, oligomycin, cyanide, and vanadate but was markedly inhibited by N-ethylmaleimide. Calmodulin failed to stimulate Ca2+-dependent ATPase and instead inhibited slightly. Unlike K+, Na+ produced a marked inhibition of the Ca2+-dependent ATPase activity, and this inhibition was associated with an 8- 10-fold decrease in the affinity of the enzyme for Ca2+. The competitive action of Na+ indicates that the Ca2+-dependent ATPase may be a site of Na+-Ca2+ antagonism in the cell membrane.

Animals↗
PubMed1982

Role of membrane integrity and cation interaction for heart sarcolemmal adenylate cyclase and Na+-K+ ATPase.

The adenylate cyclase and Na+ -K+ ATPase activities decreased on storage at 4 degrees C as well as on freezing and thawing of the rat heart sarcolemma. Treatment of the sarcolemmal fraction with phospholipase C and trypsin also depressed the adenylate cyclase and Na+ -K+ ATPase activities; the Na+ -K+ ATPase was more sensitive to these treatments than the adenylate cyclase. When the sarcolemmal enzyme activities were determined in the presence of different concentrations of some cations the adenylate cyclase activity was enhanced and the Na+ -K+ ATPase activity was depressed by monovalent cations (Na+, K+, Rb+, Cs+, Li+, and NH+4). Divalent cations such as Sr2+, Ba2+, Co2+, and Mn2+ had biphasic or no effects on the adenylate cyclase activity but inhibited the Na+ -K+ ATPase activity. Although Ca2+, Ni2+, Cd2+, Cu2+, Hg2+, and Zn2+ depressed both Na+ -K+ ATPase and adenylate cyclase activities, the degree of inhibition of these enzymes was different. These results reveal the role of membrane integrity for full expression of the adenylate cyclase and Na+ -K+ ATPase activities, whereas both monovalent and divalent cations appear to regulate sarcolemma-bound enzyme activities.

Adenylyl Cyclases↗
PubMed1982

Properties of Ca2+- or Mg2+-dependent ATPase in rat heart sarcolemma.

Rat heart sarcolemma was shown to hydrolyze ATP in the presence of Ca2+ or Mg2+; Ka values for Ca2+ and Mg2+ were in the range of 0.58-0.67 and 0.72-0.83 mM, whereas Vmax values were 33-38 and 21-28 mumol Pi/mg per hr, respectively. Both Ca2+ ATPase and Mg2+ ATPase showed low- and high-affinity sites for ATP; the Km value for the low-affinity sites for both enzyme activities was 300-325 microM, whereas Km values for high-affinity sites were 75-85 and 100-108 microM, respectively. The pattern of nucleotide hydrolysis in the presence of Ca2+ was found to be different from that with Mg2+. Although both high concentrations of ADP and Pi inhibited the enzyme activities, Mg2+ ATPase was more sensitive to ADP and less sensitive to Pi in comparison to Ca2+ ATPase. Storage of sarcolemma at about 0 degrees C showed a greater increase in ATP hydrolysis with Ca2+ than with Mg2+. The inhibitory effect of Mg2+ on Ca2+ ATPase, unlike that of Ni2+, Co2+, and Cu2+, was more than that on Mg2+ ATPase. Treatment of membranes with sodium dodecylsulfate or deoxycholate produced a greater reduction in Mg2+ ATPase than in Ca2+ ATPase. These results further support the view that Ca2+ ATPase and Mg2+ ATPase may be two separate enzymes in heart sarcolemma. It is suggested that Ca2+-dependent ATPase may be involved in opening calcium channels for the entry of calcium, whereas Mg2+ ATPase may serve as a Mg2+ pump mechanism for the efflux of magnesium from the cardiac cell.

Adenosine Triphosphatases↗
PubMed1982

Ca2+- and Mg2+-dependent ATPase activities in the deoxycholate-treated rat heart sarcolemma.

1. Isolated rat heart sarcolemma was treated with different concentrations of an ionic detergent, deoxycholate (DOC) and ATP hydrolysis in the presence of Ca2+ or Mg2+ was determined. 2. Both Ca2+-dependent ATPase and Mg2+-dependent ATPase activities were decreased in the DOC-treated membranes; however, the depression of Mg2+-dependent ATPase activity was greater than that of Ca2+-dependent ATPase. 3. The differential changes in Ca2+-dependent ATPase and Mg2+-dependent ATPase activities were apparent when incubations with DOC were carried out for different time intervals and at different temperatures. 4. In DOC-treated preparations, the Km value for Ca2+-dependent ATPase was decreased whereas that for Mg2+-dependent ATPase was increased. The half maximal velocities of the Ca2+-dependent ATPase and Mg2+-dependent ATPase enzyme reactions in the treated preparations were obtained at a DOC: membrane protein ratio of 3.0 and 0.6, respectively. 5. In the DOC-treated membranes exhibiting the half maximal velocities of enzyme reactions, the Ka value for Ca2+-dependent ATPase was drastically reduced but remained unchanged for Mg2+-dependent ATPase. 6. The DOC treatment was associated with a loss of protein as well phospholipids and resulted in changes in the ultrastructural integrity of the membrane. 7. Varying degrees of decreases in the activities of sarcolemmal adenylate cyclase. (Na+-K+)-ATPase, 5'-nucleotidase and calcium binding were seen upon DOC treatment. 8. The extent of reduction in Ca2+-dependent ATPase and Mg2+-dependent ATPase activities were also different when the membrane was treated with a non-ionic detergent, Lubrol PX. 9. These data suggest that Ca2+-dependent ATPase in heart sarcolemma is more resistant than Mg2+-dependent ATPase to detergent treatments and further indicate some differences in the properties of these enzymes.

Adenosine Triphosphatases↗
PubMed1982

Effect of adenosine in the cardiac actions of prostaglandins E2, I2, and F2 alpha.

Previous reports have demonstrated an antagonistic influence of adenosine in the biological effects of prostaglandins (PGs). We examined such a possible relationship on the isolated rat heart perfused at constant pressure and maintained at a constant heart rate. PGE2 and PGF2 alpha (2.8 X 10(-11) to 2.8 X 10(-7) M) exerted a positive inotropic influence on the heart with PGF2 alpha demonstrating the greater maximum effect. PGI2 had a negative inotropic effect only at high concentrations (2.8 X 10(-9) to 2.8 X 10(-7) M). The coronary flow was decreased by both PGE2 and PGF2 alpha. The concentrations of PGI2 employed had minimal vasoactive properties (20% increase in flow at 2.9 X 10(-7) M). Adenosine at concentrations which increased coronary flow by 24 to 63% (0.1 to 10 microM) had no influence on either the alteration in contractile force produced by PGE2 PGF2 alpha, or PGI2 or on the coronary effects of PGE2 or PGI2. Adenosine did, however, attenuate the degree of coronary flow reduction produced by PGF2 alpha administration. These results are suggestive of a selective inhibiting influence of adenosine on the coronary constricting effects of PGF2 alpha, in the isolated rat heart.

Adenosine↗
PubMed1982

Role of free radicals in catecholamine-induced cardiomyopathy.

Effects of an antioxidant, vitamin E, and a membrane stabilizing agent, zinc, were examined on the isoproterenol-induced changes in the rat myocardium. Isoproterenol treatment (80 mg/kg given over 2 days in two equal doses) caused arrhythmias and 25% mortality within 24 h of the last injection. The ultrastructural changes in the subendocardium and in focal areas of the subepicardium included swelling of mitochondria, loss of myofibrils, cell necrosis, fibrosis, and infiltration of the affected areas by polymorphonucleocytes. Both creatine phosphate and adenosine triphosphate levels were markedly decreased in hearts from isoproterenol-treated animals. Pretreatment of the animals with vitamin E (10 mg X kg-1 X day -1 for 2 weeks) or zinc (10 mg/kg ZnSO4, twice a day for 7 days) prevented these deleterious effects of isoproterenol. Animals maintained on vitamin E deficient diet for 8 weeks were found to be more sensitive to isoproterenol-induced changes and this increased sensitivity was reversed by a 2-week feeding of the animals on the normal diet coupled with vitamin E treatment. Based on the data obtained in this study it is proposed that catecholamine-induced changes may involve free radicals, which by promoting lipid peroxidation may increase membrane permeability and lead to the development of cardiomyopathy.

Animals↗
PubMed1982

Effect of cholesterol feeding and gender on the response of the isolated rat heart to hypoxia.

The effect of hypoxic perfusion on isolated hearts from male and female rats fed either a control or 2% cholesterol supplemented diet was studied. Initiation of hypoxia produced a rapid decline in rate of hearts from either male or female animals irrespective of diet regimen. However, hearts from male rats fed a control diet exhibited a significantly higher (P less than 0.01) resting rate during normoxic perfusion when compared with hearts from cholesterol-fed animals. Hypoxia also produced a rapid loss of myocardial contractile force and this effect was influenced by neither diet treatment nor animal gender. Similarly, there was little change in time to peak height of developed tension due to diet. Contracture development during hypoxic perfusion was significantly higher in hearts from female rats fed a cholesterol-supplemented diet compared with hearts from control diet fed female animals, whereas no changes were observed due to diet in hearts from male animals. Coronary resistance was increased as a result of hypoxic perfusion. In hearts from cholesterol-fed male rats this effect was substantially attenuated. Conversely, cholesterol supplementation resulted in a higher (P less than 0.05) coronary resistance in hearts from female animals either during normoxic or the early period of hypoxic perfusion. Prostacyclin synthesis as measured by immunoreactive 6-keto-PGF1 alpha efflux did not differ between hearts from either male or female rats during normoxic perfusion. Hypoxia significantly reduced 6-keto-PGF1 alpha efflux from hearts of cholesterol-fed male rats whereas it produced no effect in any other treatment category. Our results demonstrate a substantial contribution of dietary cholesterol and animal gender in the response of the isolated rat heart to hypoxic perfusion.

6-Ketoprostaglandin F1 alpha↗
PubMed1982

A possible mechanism of inotropic action of prolactin on rat heart.

Prolactin possesses positive inotropic actions in isolated heart preparations although the mechanism of this influence is not understood. Our study was designed to investigate the mechanism of this effect on the rat heart. Prolactin (50 ng/ml) produced a time-dependent increase (60%) in contractile force that reached maximum after 30 min and remained steady for a further 30 min. A similar time-dependent phenomenon was seen with 200 ng/ml prolactin although the maximum inotropic effect was reduced. Indomethacin (30 micrograms/ml) significantly reduced the inotropic effect of both prolactin concentrations although the effect of the hormone was not related to the release of 6-keto-PGF1 alpha, the prostacyclin metabolite. Propranolol (1-20 micrograms/ml) significantly reduced the positive inotropic effect of prolactin. Prolactin however had no influence on myocardial adenylate cyclase activity. Hearts that were removed from animals pretreated with 1.25 or 2.50 mg/kg reserpine did not respond to prolactin administration. It is suggested that the inotropic influence of prolactin is mediated by endogenous catecholamine liberation.

6-Ketoprostaglandin F1 alpha↗
PubMed1982

Myocardial cell damage and cardiovascular changes due to i.v. infusion of adrenochrome in rats.

In vivo effects of adrenochrome (1-32 mg/kg), an oxidation product of catecholamines, on the heart ultrastructure, ECG and blood pressure were studied in rats over a period of 60 min following a single i.v. injection of the drug. One milligram of the drug had no influence on the myocardium or the cardiovascular system, whereas maximum changes in these parameters were recorded at 32 mg/kg of adrenochrome. The maximum structural damage, reached within 5-10 min, included marked swelling of mitochondria and sarcotubular system, intracellular and perinuclear oedema, hypercontraction of myofibrils and partial separation of the intercalated disc. Ultrastructural changes in the myocardium due to 4 and 8 mg of adrenochrome were not accompanied by any cardiovascular effects and the changes were fully reversed within 60 min of the injection of the drug. However, at 16 and 32 mg/kg of adrenochrome both heart rate and blood pressure were depressed within 5 min of drug administration. At these concentrations of adrenochrome arrhythmias, mainly due to premature ventricular contractions, were also noticed. Ultrastructural and cardiovascular changes seen at these higher concentrations of adrenochrome showed only a partial recovery. The data indicates that adrenochrome-induced ultrastructural changes in the heart are due to a direct myocardial effect of the drug which may not involve haemodynamic changes and the latter are most probably a consequence of this effect. However, the present study has not been able to rule out direct vascular effects at higher concentrations of adrenochrome.

Adrenochrome↗
PubMed1981

Role of phosphatidylinositol in basal adenylate cyclase activity of rat heart sarcolemma.

The adenylate cyclase activity and phospholipid composition were determined in rat heart sarcolemma after treating the membranes with a phosphatidylinositol-specific phospholipase C. Complete hydrolysis of phosphatidylinositol in sarcolemma was associated with a marked loss of the basal adenylate cyclase activity. The recombination of the supernatant with the phosphatidylinositol-depleted membranes was found to reactivate the adenylate cyclase activity. The soluble component(s) in the supernatant, which restored the adenylate cyclase activity, was thermolabile and precipitated by ammonium sulfate. Extensive hydrolysis of phosphatidylcholine, phosphatidylethanolamine and sphingomyelin in sarcolemma with a Clostridium welchii phospholipase C treatment did not affect the basal adenylate cyclase activity. These results suggest that phosphatidylinositol anchors component(s) essential for the expression of basal adenylate cyclase activity to the myocardial cell membrane.

Adenylyl Cyclases↗
PubMed1981

Solubilization of rat heart sarcolemma 5'-nucleotidase by phosphatidylinositol-specific phospholipase C.

The influence of a phosphatidylinositol-specific phospholipase C treatment on rat heart sarcolemmal 5'-nucleotidase was investigated. Upon complete hydrolysis of all phosphatidylinositol in the sarcolemma, 75% of 5'-nucleotidase activity was found in the solubilized form. The insolubilized enzyme after this treatment has the same Km for AMP as the untreated, sarcolemmal-bound enzyme (0.04 mM), whereas the solubilized enzyme has a 40-fold increase in Km for AMP (0.16 mM). Other sarcolemmal-bound enzymes were not affected by the same treatment. Hence, the specific involvement of phosphatidylinositol in the binding of 5'-nucleotidase to the sarcolemma of the rat heart is clearly demonstrated.

5'-Nucleotidase↗