Biomedical subjects

F B Davis

Publications and source records attributed to F B Davis.

At least 55 records · Page 3Linked to original sources

PubMed1989

Retinoic acid is a modulator of thyroid hormone activation of Ca2+-ATPase in the human erythrocyte membrane.

The thyroid hormones thyroxine (T4) and 3,3',5-L-triiodothyronine (T3) stimulate plasma membrane Ca2+-ATPase (EC 3.6.1.3) activity in human erythrocytes by a mechanism independent of the cell nucleus. The current studies were conducted to determine the effect of retinoic acid on the extranuclear activation by T4 and T3 of Ca2+-ATPase in the human red cell. The retinoid inhibited basal and T4-stimulatable activity of that enzyme in a dose-dependent manner. At the highest tested concentration (10(-6) M), retinoic acid inhibited basal enzyme activity by 25% and T4-stimulated activity by 72%. A concentration as low as 5 x 10(-10) M retinoic acid shifted the dose-response curve of both T4 and T3 so that the concentration of each associated with maximal enzyme stimulation was 10(-9) M instead of 10(-10) M. Retinoic acid displaced [125I]T4 binding to red cell membranes as effectively as unlabeled T4. Retinol failed to influence either basal or T4-stimulated enzyme activity or to displace T4 binding. These results indicate that retinoic acid can partially block the T4 and T3 stimulation of Ca2+-ATPase in human red cell membranes and suggest a physiologic role for the retinoid as a modulator of this peripheral action of thyroid hormone. They suggest that the red cell membrane is an important site of action for this active retinoid.

Calcium-Transporting ATPases↗

PubMed1989

Thyroid hormone regulation of membrane Ca2(+)-ATPase activity.

The Ca2(+)-ATPase of plasma membranes from a variety of tissues is subject to stimulation in vitro, and apparently in vivo, by physiological concentrations of iodothyronines regarded as biologically active in other bioassay systems. This calmodulin-dependent action of thyroid hormone is nongenomic, that is, directly on the cell membrane and independent of the cell nucleus. In the case of human erythrocyte Ca2(+)-ATPase, this assay of thyroid hormone bioactivity is attractive as an in vitro, readily-studied model of hormone action in a human cell. Enzyme activity is paralleled, as expected, by changes in calcium pump activity. Thyroid hormone action in this system is subject to modulation by glucose and by a variety of compounds which, like iodothyronines, are hydrophobic. The mechanism of thyroid hormone action on membrane Ca2(+)-ATPase involves, at least in part, membrane lipids, including components of the phosphatidylinositol cycle. The physiologic role of thyroid hormone action on cell membrane Ca2(+)-ATPase is speculative. In plasma membranes of nonexcitable and excitable tissues, ambient thyroid hormone may set basal activity of Ca2(+)-ATPase or magnitude of the enzymatic response to calmodulin Ca2+.

Animals↗

PubMed1988

Abnormal response to calmodulin in vitro of dystrophic chicken muscle membrane Ca2+-ATPase activity.

A skeletal muscle membrane fraction enriched in sarcoplasmic reticulum (SR) contained Ca2+-ATPase activity which was stimulated in vitro in normal chickens (line 412) by 6 nM purified bovine calmodulin (33% increase over control, P less than 0.001). In contrast, striated muscle from chickens (line 413) affected with an inherited form of muscular dystrophy, but otherwise genetically similar to line 412, contained SR-enriched Ca2+-ATPase activity which was resistant to stimulation in vitro by calmodulin. Basal levels of Ca2+-ATPase activity (no added calmodulin) were comparable in muscles of unaffected and affected animals, and the Ca2+ optima of the enzymes in normal and dystrophic muscle were identical. Purified SR vesicles, obtained by calcium phosphate loading and sucrose density gradient centrifugation, showed the same resistance of dystrophic Ca2+-ATPase to exogenous calmodulin as the SR-enriched muscle membrane fraction. Dystrophic muscle had increased Ca2+ content compared to that of normal animals (P less than 0.04) and has been previously shown to contain increased levels of immuno- and bioactive calmodulin and of calmodulin mRNA. The calmodulin resistance of the Ca2+-ATPase in dystrophic muscle reflects a defect in regulation of cell Ca2+ metabolism associated with elevated cellular Ca2+ and calmodulin concentrations.

Animals↗

PubMed1988

Calcium channel blocker inhibition of the calmodulin-dependent effects of thyroid hormone and milrinone on rabbit myocardial membrane Ca2+-ATPase activity.

The Ca2+-ATPase activity of rabbit myocardial membranes is stimulated in vitro by L-thyroxine and by milrinone, a bipyridine. These effects are concentration dependent and calmodulin requiring. The calcium channel blockers nifedipine and verapamil have been reported to have anti-calmodulin effects in other assay systems. In this study we have examined the effects of nifedipine and verapamil on rabbit myocardial membrane Ca2+-ATPase activity, in the absence (basal activity) and presence of exogenous L-thyroxine (T4), 10(-10) M, and milrinone, 10(-7) M. Basal enzyme activity was inhibited by a minimum of 10(-6) M nifedipine (IC50 of 3.4 X 10(-5) M) and 10(-5) M verapamil (IC50 of 1.5 X 10(-4) M). Both calcium antagonists inhibited enzyme stimulation by T4 and milrinone, with half-maximal inhibition of T4 and milrinone effects, respectively, at 2.9 X 10(-5) M and 9.0 X 10(-6) M nifedipine and 3.0 X 10(-5) M and 5.2 X 10(-5) M verapamil. The addition of exogenous purified calmodulin, 40 ng/micrograms membrane protein, in the presence of 10(-5) M nifedipine or verapamil restored T4-stimulated enzyme activity. Nifedipine and verapamil, each at a concentration of 10(-6) M, significantly inhibited binding of radioiodinated calmodulin to rabbit heart membranes in vitro. These studies provide evidence that nifedipine and verapamil have an anti-calmodulin effect in this myocardial enzyme system. Through interaction with calmodulin, the channel blockers inhibit thyroid hormone and milrinone stimulation of myocardial membrane Ca2+-ATPase.

Animals↗

PubMed1988

Early abnormal development of calmodulin gene expression and calmodulin-resistant Ca2+-ATPase activity in avian dystrophic muscle.

We have reported previously that the pectoralis muscle from three month-old dystrophic chickens with signs of myopathy exhibits increased calmodulin content, elevated calmodulin-specific mRNA (Biochem. Biophys. Res. Commun. 137:507-512, 1986), and reduced sarcoplasmic reticulum (SR) Ca2+-ATPase activity in response to calmodulin exposure in vitro (Clin. Res. 34: 725A, 1986). To determine the early time sequence for development of these abnormalities, we have studied muscle from embryos and post-hatched chickens at various ages. Quantitated by dot blot analysis, there was an approximate two-fold increase in calmodulin-specific mRNA in dystrophic muscle as early as 13 days ex ovo which was maintained throughout development up to three months ex ovo. Similarly, Ca2+-ATPase activity measured in SR membranes from chickens as early as 13 days post-hatch was also found to be resistant to stimulation in vitro by exogenous calmodulin, whereas the enzyme from normal muscle was calmodulin-stimulable. These findings suggest that the genetic lesion expressed in the avian dystrophic animal model involves the loss of normal control of intracellular calcium metabolism early in the maturation of the affected musculature and prior to appearance of disease signs.

Animals↗

PubMed1988

Analogue-specific action in vitro of atrial natriuretic factor on human red blood cell Ca2+-ATPase activity.

Specific atrial natriuretic factor (ANF) analogues have been found to have inhibitory activity in vitro in a calmodulin-dependent, human red blood cell membrane Ca2+-adenosine triphosphatase (ATPase) model. Studied at 10(-8) to 10(-6) M concentrations, atriopeptin I (residues 127-147 of rat prepro-ANF sequence) and atriopeptin III (residues 127-150) progressively inhibited Ca2+-ATPase activity by up to 20% (p less than 0.001). This degree of inhibition was consistent with activities of other (calmodulin-independent) enzyme inhibitors in this model. Therefore, the C-terminal Phe-Arg-Tyr sequence (residues 148-150) is unnecessary for atriopeptin action on Ca2+-ATPase. Human and rat atrial peptides with amino acids 123-150 were inactive, indicating that the 123-126 sequence (Ser-Leu-Arg-Arg) must be cleaved to activate atriopeptins in this system. Human ANF fragment 129-150 also had no effect on Ca2+-ATPase, defining the importance of residues 127-128 (Ser-Ser) proximal to the disulfide bridge (joining 129 to 145). The addition of purified calmodulin to red blood cell membranes in the presence of inhibitory ANF did not restore Ca2+-ATPase activity to normal levels, indicating that the ANF effect on this enzyme is calmodulin-independent. Atriopeptin I and atriopeptin III had no effect on red blood cell Na+, K+-ATPase activity in vitro. Thus, the structure-activity relationships of ANF analogues in this novel human cell membrane model are highly specific. Although the inhibitory action of ANF analogues on Ca2+-ATPase, a calcium pump-associated enzyme, may be unique to the red blood cell, the calcium dependence of the gluconeogenic effects of ANF in the kidney would be supported by inhibition of this ATPase.

Amino Acid Sequence↗

PubMed1987

Action of long-chain fatty acids in vitro on Ca2+-stimulatable, Mg2+-dependent ATPase activity in human red cell membranes.

Human red cell membrane Ca2+-stimulatable, Mg2+-dependent adenosine triphosphatase (Ca2+-ATPase) activity and its response to thyroid hormone have been studied following exposure of membranes in vitro to specific long-chain fatty acids. Basal enzyme activity (no added thyroid hormone) was significantly decreased by additions of 10(-9)-10(-4) M-stearic (18:0) and oleic (18:1 cis-9) acids. Methyl oleate and elaidic (18:1 trans-9), palmitic (16:0) and lauric (12:0) acids at 10(-6) and 10(-4) M were not inhibitory, nor were arachidonic (20:4) and linolenic (18:3) acids. Myristic acid (14:0) was inhibitory only at 10(-4) M. Thus, chain length of 18 carbon atoms and anionic charge were the principal determinants of inhibitory activity. Introduction of a cis-9 double bond (oleic acid) did not alter the inhibitory activity of the 18-carbon moiety (stearic acid), but the trans-9 elaidic acid did not cause enzyme inhibition. While the predominant effect of fatty acids on erythrocyte Ca2+-ATPase in situ is inhibition of basal activity, elaidic, linoleic (18:2) and palmitoleic (16:1) acids at 10(-6) and 10(-4) M stimulated the enzyme. Methyl elaidate was not stimulatory. These structure-activity relationships differ from those described for fatty acids and purified red cell Ca2+-ATPase reconstituted in liposomes. Thyroid hormone stimulation of Ca2+-ATPase was significantly decreased by stearic and oleic acids (10(-9)-10(-4) M), but also by elaidic, linoleic, palmitoleic and myristic acids. Arachidonic, palmitic and lauric acids were ineffective, as were the methyl esters of oleic and elaidic acids. Thus, inhibition of the iodothyronine effect on Ca2+-ATPase by fatty acids has similar, but not identical, structure-activity relationships to those for basal enzyme activity. To examine mechanisms for these fatty acid effects, we studied the action of oleic and stearic acids on responsiveness of the enzyme to purified calmodulin, the Ca2+-binding activator protein for Ca2+-ATPase. Oleic and stearic acids (10(-9)-10(-4) M) progressively inhibited, but did not abolish, enzyme stimulation by calmodulin (10(-9) M). Double-reciprocal analysis of the effect of oleic acid on calmodulin stimulation indicated noncompetitive inhibition. Addition of calmodulin to membranes in the presence of equimolar oleic acid restored basal enzyme activity. Oleic acid also reduced 125I-calmodulin binding to membranes, but had no effect on the binding of [125I]T4 by ghosts. The mechanism of the decrease by long chain fatty acids of Ca2+-ATPase activity in situ in human red cell ghosts thus is calmodulin-dependent and involves reduction in membrane binding of calmodulin.

Ca(2+) Mg(2+)-ATPase↗

PubMed1987

Competition of milrinone, a non-iodinated cardiac inotropic agent, with thyroid hormone for binding sites on human serum prealbumin (TBPA).

Milrinone [2-methyl-5-cyano-(3,4'-bipyridin)-6(1H)-one] is a positive cardiac inotropic agent recently shown to have thyromimetic activity in vitro in a rabbit myocardial membrane Ca2+-ATPase system [K. M. Mylotte et al., Proc. natn. Acad. Sci. U.S.A. 82, 7974 (1985)]. In the present studies, milrinone was examined for activity as an inhibitor of iodothyronine binding by human serum thyroid hormone transport proteins, thyroxine-binding globulin (TBG), prealbumin (TBPA) and albumin. Polyacrylamide gel electrophoresis at pH 9.0 of sera equilibrated with [125I]thyroxine showed that milrinone competed with L-thyroxine (T4) for binding sites on TBPA (10 and 100 microM milrinone caused 61 and 73% reductions, respectively, in T4 binding to TBPA, P less than 0.01); T4 displaced from TBPA was bound by TBG and albumin. Comparable reductions in T4 binding to TBPA were observed in electrophoretic studies conducted at pH 7.4. Binding of triiodo-L-thyronine (T3) to TBPA was electrophoretically confirmed and shown to be decreased in the presence of milrinone. Electrophoresis of purified TBPA also demonstrated that [14C]milrinone co-migrated with this transport protein and that milrinone displaced tracer T4 from TBPA. Amrinone, the 2-H-5-NH2 analog of milrinone, had less than 5% of the activity of milrinone as an inhibitor of T4 binding in electrophoretic studies. Scatchard analysis of T4 and milrinone binding to purified TBPA, measured by equilibrium dialysis, showed two classes of binding sites, with association constants, respectively, of 6.1 X 10(7) M-1 and 1.6 X 10(6) M-1 for T4, and 1.7 X 10(6) M-1 and 8.9 X 10(2) M-1 for milrinone. Computer graphic modeling of the binding of milrinone to the T4 site in the crystal structure of TBPA showed that milrinone best occupied this site when the substituted bipyridine ring overlapped the phenolic ring of T4. In this orientation the 5-cyano group, which has an electronegativity similar to that of iodine, occupied the same volume as the 5'-iodine of T4. The 5-amino group of amrinone lacks these characteristics. In this orientation, the keto function of milrinone overlapped the T4 4'-hydroxyl and could participate in similar intermolecular interactions. Thus, milrinone, a non-iodinated bipyridine, and thyroid hormone share structural and biochemical homologies and compete for the same binding site on TBPA.

Amrinone↗

PubMed1987

Donor age-dependent decline in response of human red cell Ca2+-ATPase activity to thyroid hormone in vitro.

The effect of increasing donor age on the susceptibility of human red blood cell Ca2+-ATPase activity to stimulation in vitro by thyroid hormone was studied in 26 normal subjects, aged 15-81 yr. Basal enzyme activity (no added thyroid hormone) was unaffected by donor age. Group analysis, young (less than or equal to 50 yr) vs. elderly (greater than 60 yr old), revealed a 23% decrease in responsiveness of the enzyme to L-T4 (P less than 0.001). Regression analysis confirmed an age-dependent decline in thyroid hormone stimulability of Ca2+-ATPase [r = -0.42 (T4 effect) and -0.38 (T3 effect); P less than 0.01]. Red cell membrane Na,K-ATPase activity was not affected by donor age. Plasma T4 and T3 concentrations in these normal subjects also did not change with age. Possible contributions of the following mechanisms to this age-correlated change in enzyme activity were examined: altered responsiveness to calmodulin of membrane Ca2+-ATPase; membrane content of endogenous calmodulin, endogenous plasma T4 and T3 concentrations, and plasma glucose concentrations. Calmodulin responsiveness is required for iodothyronine action on the enzyme, but the calmodulin responsiveness of cells from elderly donors was not significantly different from that of cells from younger donors (P greater than 0.10). There was no relationship between membrane immunoassayable calmodulin and donor age or membrane calmodulin and Ca2+-ATPase activity. There were positive correlations between donor plasma T4 level and basal enzyme activity (P less than 0.05) and between donor plasma T3 concentration and hormone-responsive Ca2+-ATPase (P less than 0.01), but these did not contribute to the age effect. Plasma glucose previously was found to modulate red cell Ca2+-ATPase activity, but did not correlate with decreased hormone responsiveness of the enzyme in elderly donors. In conclusion, we found that the susceptibility of human red cell Ca2+-ATPase to in vitro thyroid hormone stimulation declined significantly with advancing donor age. Several possible calmodulin-dependent mechanisms for this age-dependent change were excluded, and thus, we postulate that the altered hormone sensitivity of the enzyme is membrane phospholipid mediated.

Adolescent↗

PubMed1987

Bepridil and cetiedil reversibly inhibit thyroid hormone stimulation in vitro of human red cell Ca2+-ATPase activity.

Thyroid hormone (10(-11) to 10(-10) M) stimulates plasma membrane Ca2+-ATPase activity in vitro in various tissues, including the human red cell (RBC), by a calmodulin-requiring mechanism. Bepridil and cetiedil are Ca2+ antagonists with an intracellular (calmodulin-antagonist) site of action, as well as an effect on the calcium channel in excitable tissues. We have studied the actions of bepridil and cetiedil on Ca2+-ATPase in a channel-free membrane (RBC) to determine effectiveness of these agents as inhibitors of thyroid hormone action on the enzyme. Dose-response studies showed that thyroid hormone stimulation of Ca2+-ATPase activity in vitro was significantly inhibited by as little as 2 x 10(-5) M bepridil and cetiedil. IC50 values of bepridil and cetiedil for thyroid hormone response of the enzyme were 5 x 10(-5) and 2 x 10(-5) M, respectively, whereas IC50s of these agents for enzyme activity in the absence of thyroid hormone were both 10(-4) M. Progressive addition of purified rat testis calmodulin in vitro (10-150 ng calmodulin/mg membrane protein) restored hormone responsiveness in the presence of bepridil and cetiedil. Binding of labeled thyroid hormone by RBC membranes was unaffected by bepridil and cetiedil (up to 2 x 10(-4) M). Thus, bepridil and cetiedil are Ca2+ antagonists that reversibly inhibit thyroid hormone action on human RBC Ca2+-ATPase by a calmodulin-dependent mechanism. Thyroid hormone effect on Ca2+-ATPase is more susceptible to bepridil and cetiedil inhibition than is basal enzyme activity.

Azepines↗

PubMed1987

Water excretion in the elderly.

Osmoreceptor sensitivity is enhanced in healthy elderly subjects and AVP secretion is increased, relative to that of younger subjects, when plasma osmolality rises. Increased AVP secretion/unit increase in plasma tonicity reflects a decrease in collecting tubule sensitivity to AVP by an as yet unknown mechanism in the aged kidney. This change in sensitivity is not completely offset by increased ADH release, so that maximum Uosm achievable under hydropenic conditions (concentrating ability) is reduced in the elderly. CH2O in older subjects decreases in proportion to the fall in GFR; thus, CH2O is intact in older subjects with preserved GFR. In subjects with age-related reductions in GFR, minimal Uosm achievable is usually less than 100 mOsm per kg H2O and thus usually sufficient to meet the demands of solute-free water intake so that plasma hypo-osmolarity does not result. Increasing exposure of the elderly to pharmacologic agents that reduce CH2O is primarily responsible for the impression that aged patients are at increased risk for hyponatremia.

Adult↗

PubMed1986

Abnormal expression of the calmodulin gene in muscle from the dystrophic chicken.

Compared to that of genetically-related normal chickens, pectoralis muscle from the dystrophic chicken contained increased calmodulin measured by radioimmunoassay. Determined by the dot blot procedure, expression of the calmodulin gene was enhanced in muscle from affected animals. The bioactivity of the gene product was normal. Together with previous studies reporting increased cell Ca2+ content in dystrophic muscle, the current findings of increased sarcoplasmic calmodulin suggest the latter is a cellular response to defective Ca2+ transport at the level of cell efflux or intracellular organelle (sarcoplasmic reticulum) uptake.

Animals↗

PubMed1986

Urinary diluting capacity in elderly diabetic subjects.

Urinary concentrating ability declines with normal human aging but diluting capacity has been less well studied as a function of age. We studied free water clearance (CH2O) in a group of Type II diabetic patients, aged 47 to 70 years. Conventional water load testing in sulfonylurea-treated diabetic patients revealed preservation of diluting capacity (ability to achieve Uosm less than 100 mOsm/kg) in subjects greater than 60 years of age. Tolazamide permitted expression of normal CH2O; chlorpropamide administration predictably reduced CH2O relative to tolazamide by 58% in subjects greater than 60 years. Free water clearance (CH2O) and glomerular filtration rate (GFR) were comparable in elderly subjects (greater than 60 years) and those less than or equal to 60 years. Normal CH2O in these diabetic patients was explained by persistently normal GFR in the study group. Osmolar clearance (Cosm) was insignificantly lower in diabetics greater than 60 years. Thus, a fall in CH2O in elderly diabetic patients is not a factor contributing to clinical hypoosmolar states encountered in this population.

Aged↗

PubMed1986

Effects of hyperthyroidism and hypothyroidism on human red blood cell Ca2+-ATPase activity.

Erythrocyte Ca2+-ATPase activity was measured in membrane ghosts from hyperthyroid and hypothyroid patients and compared with that in normal subjects. Basal enzyme activity was significantly increased in the hyperthyroid group and decreased in the hypothyroid group. In vitro responsiveness of the enzyme to calmodulin, the activator protein for Ca2+-ATPase, and to T4 was decreased in both hyper- and hypothyroid ghosts. Although the membrane content of endogenous calmodulin was low in hypothyroidism, this is an unlikely explanation for the low basal activity of Ca2+-ATPase. A limited number of patients from each group were restudied when they were euthyroid after 8 or more months of treatment. Basal, T4-stimulated, and calmodulin-stimulated Ca2+-ATPase activities returned to normal, with the exception of calmodulin responsiveness in the membranes from previously hypothyroid patients. This decreased responsiveness may have been a reflection of either increased patient age or underlying chronic (non-thyroidal) illness. This study provides clinical confirmation of previously reported in vitro studies suggesting that the set-point of Ca2+-ATPase activity is, in part, a function of endogenous thyroid hormone levels.

Adult↗

PubMed1986

Differential activities of tolbutamide, tolazamide, and glyburide in vitro on rabbit myocardial membrane Ca2+-transporting ATPase activity.

At clinically achievable concentrations (10(-9) to 5 X 10(-6) M), tolbutamide and tolazamide are in vitro inhibitors of Ca2+-transporting ATPase activity in sarcolemma-enriched rabbit myocardial membranes (sulfonylurea IC50, 10(-7) M). Thyroid hormone stimulation of this calcium pump-associated enzyme in vitro has been previously reported; in our study, this hormonal action was shown to be inhibited by tolbutamide and tolazamide. In contrast to these two sulfonylureas, glyburide (up to 5 X 10(-6) M) had no effect on basal or thyroid hormone-stimulable Ca2+-ATPase activity in vitro. Studies of binding of radiolabeled purified calmodulin to heart membranes showed that tolbutamide and tolazamide inhibited this interaction, whereas glyburide had no effect on calmodulin binding. Addition of purified calmodulin (5-40 ng/micrograms membrane protein) to myocardial membranes incubated with 10(-7) M tolbutamide or tolazamide restored Ca2+-ATPase activity and thyroid hormone responsiveness of the enzyme. Inhibition by tolbutamide and tolazamide of myocardial sarcolemmal Ca2+-ATPase is a mechanism by which these two sulfonylureas may at least transiently raise resting sarcoplasmic Ca2+ concentration. This effect of sulfonylureas on Ca2+-ATPase is not expressed in the presence of the benzamide side chain of glyburide. The inhibitory action of certain sulfonylureas on Ca2+-ATPase is mediated by interference of the agents with the binding of calmodulin to cardiac membranes.

Animals↗

PubMed1985

Milrinone and thyroid hormone stimulate myocardial membrane Ca2+-ATPase activity and share structural homologies.

We have recently shown that thyroid hormone in physiological concentrations stimulates sarcolemma-enriched rabbit-myocardial-membrane Ca2+-ATPase in vitro. In this study, milrinone [2-methyl-5-cyano-(3,4'-bipyridin)-6(1H)-one], a cardiac inotropic agent, was thyromimetic in the same system. At clinically achievable concentrations (50-500 nM), milrinone significantly stimulated membrane Ca2+-ATPase in vitro. This action was antagonized by W-7 [N-(6-aminohexyl)-5-chloro-1-naphthalenesulfonamide], an agent that also blocks thyroid hormone action on the Ca2+-ATPase, at concentrations as low as 5 microM. Progressive additions of milrinone to membranes incubated with a fixed concentration of thyroxine (0.10 nM) or triiodothyronine resulted in a progressive obliteration of the thyroid hormone effect on Ca2+-ATPase. Amrinone [5-amino-(3,4'-bipyridin)-6(1H)-one], the parent bipyridine of milrinone, had no effect on myocardial Ca2+-ATPase activity. X-ray crystallographic analysis of milrinone and amrinone revealed structural homologies between the phenolic ring of thyroxine and the substituted ring of milrinone, whereas amrinone did not share these homologies. The mechanism(s) of the inotropic actions of thyroxine and of milrinone is not clearly understood, but these observations implicate Ca2+-ATPase, a calcium pump-associated enzyme, as one mediator of the effects on the heart of these two compounds.

Aminopyridines↗

PubMed1985

Diagnosis of hyperparathyroidism.

Availability of immunoassays for specific regions of the parathyroid hormone (PTH) molecule allows discrimination with a high level of surety between primary hyperparathyroidism and tumoral hypercalcemic states associated with circulating PTH-like substances. Assay for intact, N-terminal PTH currently has the highest discriminant function. Prostaglandin-dependent and osteoclast-activating factor-mediated hypercalcemic states associated with neoplasia have suppressed serum PTH levels. PTH-like substances are detected by immunoassays, but in the intact, N-terminal system they are seen as normal-range or low values. The frequency with which any tumor produces only authentic PTH is very low. The serum chloride:phosphate ratio has limited clinical utility in distinguishing tumoral hypercalcemia from hyperparathyroid hypercalcemia, and measurements of nephrogenous cyclic AMP do not distinguish between the effects of circulating authentic PTH and PTH-like substances elaborated by tumors. Additional measures that, in the future, may help to distinguish between parathyroid and tumoral hypercalcemias include quantitative bone biopsy histomorphometry and in vitro bioassays for PTH activity in the separate plasma fractions, obtained by gel filtration, in which PTH and PTH-like substances are found.

Adenoma↗

PubMed1985

The effect of in vivo glucose administration on human erythrocyte Ca2+-ATPase activity and on enzyme responsiveness in vitro to thyroid hormone and calmodulin.

To characterize endogenous control mechanisms for human erythrocyte membrane Ca2+-ATPase ("calcium pump") activity, we studied the effect of changes in blood glucose concentration in vivo within the physiologic range on Ca2+-ATPase activity in red cells. Red cells obtained in the course of induced hyperglycemia were also studied to determine susceptibility of membrane Ca2+-ATPase to stimulation in vitro by thyroid hormone and calmodulin, both of which have been shown previously to enhance Ca2+-ATPase activity. Oral glucose administration (75 g) to eight healthy, adult subjects induced predictable increases in concentrations of blood glucose and immunoreactive insulin. Basal levels of activity of Ca2+-ATPase in red cells obtained after glucose ingestion fell 55% (P less than 0.025) by 30 min after glucose with recovery of enzyme activity to levels not significantly different from basal by 60 min. Activity of red cell Ca2+-ATPase at time zero was significantly stimulated in vitro by thyroxine (T4, 10(-10) M), triiodo-L-thyronine (T3, 10(-10) M), and calmodulin (100 ng/mg membrane protein). In vivo glucose administration led to depression of red cell enzyme responsiveness in vitro to T4 and T3; recovery from this effect did not occur by 120 min after oral administration of glucose. Calmodulin responsiveness of the enzyme in vitro was less significantly reduced in red cells obtained after glucose ingestion. Intravenous (i.v.) glucose administration (20 g) to five subjects also led to decreased basal enzyme activity (61% of fasting level at 20 min). A significant decrease in response of enzyme to T4 was achieved by 8 min after glucose administration (P less than 0.02), with recovery by 60 min.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗