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Biomedical subjects

D D Fanestil

Publications and source records attributed to D D Fanestil.

At least 73 records · Page 4Linked to original sources
PubMed1982

Spironolactone: a glucocorticoid agonist or antagonist?

Spironolactone displaces 3H-dexamethasone from HTC cell nuclei and inhibits the induction of tyrosine aminotransferase (TAT) activity by dexamethasone in HTC cells. Spironolactone alone fails to increase TAT activity in HTC cells or in the liver of adrenalectomized rats. These actions indicate that spironolactone lacks glucocorticoid agonist activity but can be an antagonistic to the actions of glucocorticoid as well as mineralocorticoid steroid hormones.

Adrenalectomy↗
PubMed1981

Effects of inhibitors of F0-F1 proton-translocating ATPase on urinary acidification.

A proton pump in diverse biological systems consists of two structural units with separate but integrated functions, the F0-F1-ATPase. We tested by chemical perturbation the possibility that such a proton pump might be involved in urinary acidification conducted by urinary epithelia (UF0-UF1-ATPase). Tyrosine-reactive chemicals and N,N'-dicyclohexylcarbodiimide, known to block the proton channel unit (F0), also inhibited urinary acidification, as measured by the reverse short-circuit current (RSCC) in urinary bladders from toads and turtles. Since these chemicals were equally effective under aerobic and anaerobic conditions, the inhibition appears to occur directly on UF0 rather than on mitochondria. In contrast, an inhibitor of F0-F1-ATPase, oligomycin, only inhibited aerobic RSCC and was ineffective on anaerobic RSCC. Thus, oligomycin appears to inhibit mitochondrial F0-F1 rather than any UF0+UF1. Another inhibitor of the F1-ATPase unit, N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline, was without effect on RSCC at a concentration (0.3 mM) that produced over 50% inhibition of short-circuit current. These results support the concept that acidifying urinary epithelia contain a plasma membrane proton channel, UF0, but that UF1, if it exists, is unique in that it is resistant to known inhibitors.

Adenosine Triphosphatases↗
PubMed1980

Binding of the chymotrypsin substrate, tryptophan methyl ester, by rat alpha-fetoprotein.

We studied how tryptophan methyl ester and related compounds inhibit binding of estrone to rat alpha-fetoprotein and find that: (a) like chymotrypsin, alpha-fetoprotein binds tryptophan esters with higher affinity than tryptophan or its amides; (b) the affinity of alpha-fetoprotein for tryptophan methyl ester is 3.7 . 10(-4) M, which is close to the affinity of chymotrypsin (10(-4) M); (c) alpha-fetoprotein binding of tryptophan methyl ester is stereoselective and pH dependent. All of these observations suggest that there is a specific interaction between alpha-fetoprotein and the chymotrypsin substrate, tryptophan methyl ester, and that rat alpha-fetoprotein contains a site with some structural similarities to the catalytic site in chymotrypsin. Since we also find that tryptophan methyl ester is a competitive inhibitor of estrone binding to alpha-fetoprotein, it is possible that the protease substrate binding site on alpha-fetoprotein is spatially close to the estrone binding site.

Animals↗
PubMed1980

Vanadate: non-selective inhibition of transepithelial transport of Na+, H+ and water.

In the isolated urinary bladder of the toad, 10(-5)-10(-4)M orthovanadate produces inhibition of the active transport of Na+ and H+ ions as well as of antidiuretic hormone-mediated osmotic flow of water. Since transport of H+ ions and osmotic water flow are not inhibited when (Na+ + K+)-ATPase is inhibited by ouabain, biological actions of vanadate are not necessarily related to inhibition of (Na+ + K+)-ATPase.

Animals↗
PubMed1980

Covalent modification and inhibition of an epithelial sodium channel by tyrosine-reactive reagents.

This study sought to elucidate the molecular mechanism involved in the Na+ entry across the apical membrane of the urinary bladder of the toad. Na+ transport, as measured by short-circuit current (SCC), was irreversibly inhibited by three tyrosine-specific reagents: N-acetylimidazole (ID50, 4.6 x 10(-2)M), tetranitromethane (1.8 x 10(-4) M), and 7-chloro-4-nitrobenzo-2-oxa-1,3-diazole (NBD-Cl; 3.4 x 10(-5) M). The functional specificity of NBD-Cl to block Na+ entry via Na+ channels was attested by the following: 1) NBD-Cl produced comparable inhibition of SCC and Na+ influx under aerobic and anaerobic conditions; 2) amphotericin B produced complete recovery of inhibited SCC; 3) vasopressin increased SCC only in proportion to the uninhibited SCC; 4) Km for Na+ was not changed; and 5) the half time for the inhibition varied as a function of amiloride concentration or pharmacologic activity of its analogues. On the basis of the above findings, these tyrosine-specific reagents are believed to be useful chemical probes for the identification and characterization of Na+ channel protein.

4-Chloro-7-nitrobenzofurazan↗
PubMed1979

Digoxin overdose. Limitations of hemoperfusion-hemodialysis treatment.

We used charcoal hemoperfusion coupled with hemodialysis to treat a woman with massive digoxin ingestion complicated by hyperkalemia. Although dialysis controlled the serum potassium levels, hemoperfusion removed less than 1% of the total ingested dose. Hemoperfusion has a relatively minor impact on digoxin elimination and remains of unproved value in the therapy for digoxin overdose.

Adult↗
PubMed1979

Metabolic dependence of the offset of antidiuretic hormone-induced osmotic flow of water across the toad urinary bladder.

The elevated osmotic permeability to water induced by antidiuretic hormone (ADH) in the isolated urinary bladder of the toad is rapidly reversed by removal or washout of the ADH. This return to normal water permeability is delayed by the suppression of production of metabolic energy by any of three maneuvers: (i) low temperature (2 degrees C); (ii) inhibition of oxidative phosphorylation (10 mM azide or 0.5 mM 2,4 dinitrophenol); or (iii) inhibition of glycolysis (10 mM iodoacetate or 10 mM 2-deoxyglucose). Moreover exposure to cytochalasin B, 2.1 X 10(-5) M, either before or after initiation of the hormonal effect also delays the return of water permeability to normal following removal of ADH. When considered within constraints imposed by models which predict ADH's action on water permeability to be either via modulation of the fluidity of lipids in the membrane or via the figuration of proteins ("pores") in the lipid membrane, these observations on the inhibition of the reversal of ADH stimulation of water flow are more consistent with the protein (pore) theory and place limitations on the mechanisms by which proteins in such pores can return to the resting or impermeable state.

Animals↗
PubMed1979

Inhibition of short-circuit current by triaminopyrimidine in isolated toad urinary bladder.

The organic cation 2,4,6-triaminopyrimidine (TAP) produced inhibition of short-circuit current (SCC) when added to either the mucosal or serosal surface of the isolated urinary bladder of the toad. Fifty percent inhibition was produced by 10(-3) M TAP in the mucosal solution at pH 6.8 when the mucosal [Na+] was 113 mM. The actions of TAP resemble those produced by amiloride in several ways: a) inhibition of SCC by mucosal application is rapid; b) the mucosal inhibition is fully reversible; c) high concentrations in the serosal solutions produce irreversible inhibition; and d) the concentration required to produce 50% inhibition from the mucosal side is reduced when mucosal [Na+] is reduced. It is postulated that mucosal application of TAP and amiloride inhibit short-circuit current in high-resistance epithelia via action at a common locus.

Amiloride↗
PubMed1978

Microviscosity of mucosal cellular membranes in toad urinary bladder: relation to antidiuretic hormone action on water permeability.

The microviscosity of cellular membranes (or membrane fluidity) was measured in suspensions of single mucosal cells isolated from the urinary bladder of the toad, Bufo marinus, by the technique of polarized fluorescence emission spectroscopy utilizing the hydrophobic fluorescent probe, perylene. At 23 degrees C, 5 mM dibutyryl cyclic 3',5'-AMP decreased the apparent microviscosity of the cell membranes from 3.31 to 3.07 P, a minimum decrease of 7.3% (P less than 0.001) with a physiological time course. Direct visualization of the cell suspension indicated that 98% of the cells were viable, as indicated by Trypan Blue dye exclusion. The fluorescent perylene could be seen only in plasma membranes, suggesting that the measured viscosity was that of plasma membrane with little contribution from the membranes of cellular organelles. Addition of antidiuretic hormone to intact hemibladders stained with perylene produced changes in fluorescence consistent with a similar 7% decrease in apparent microviscosity with a physiological time course. However, finite interpretation of the findings in intact tissue cannot be made because the location and the fluorescent lifetime of the probe could only be conducted on the isolated cells. Comparison with previously determined relationships between water permeability and microviscosity in artificial bilayers suggests that the 7% (a lower limit) decrease in microviscosity would produce only a 6.5% increase in water permeability.

Animals↗
PubMed1978

Inhibition by protease inhibitors of binding of adrenal and sex steroid hormones.

Binding of steroid hormones is inhibited by protease inhibitors and substrates. The protease inhibitors phenylmethyl sulphonylfluoride, tosyl-lysine chloromethyl ketone, and tosylamide-phenylethyl-chloromethyl ketone and the protease substrates tosyl arginine methyl ester and tryptophan methyl ester eliminate specific binding of aldosterone, dexamethasone, dihydrotestosterone, estrogen, and progesterone to their respective receptors. These protease inhibitors and substrates also inhibit binding of progesterone to the 20,000 molecular weight mero-receptor formed from the progesterone receptor in chick oviduct. The binding of estradiol to rat alpha-fetoprotein is inhibited by the protease inhibitors and substrates but not by tryptophan or tryptophan amide, indicating the importance of an ester structure in the inhibition of steroid binding. Our results suggest that all steroid hormone receptors have a site with both common structural features and a role in the regulation of steroid hormone binding.

Aldosterone↗
PubMed1978

Stimulation of urinary acidification by aldosterone and inhibitors of RNA and protein synthesis.

Urinary acidification by the urinary bladder of the toad (Bufo marinus) was stimulated, relative to control, by the in vitro addition of aldosterone (10(-7) M), actinomycin D (20 microgram/ml), puromycin (80 microgram/ml) or cycloheximide (5 microgram/ml). The action of the inhibitors of RNA or protein synthesis was not additive with that of aldosterone. This is opposite to the situation with Na+ transport, where the stimulation by aldosterone is abolished by the same concentrations of these inhibitors. That all agents enhanced urinary acidification was verified by: (i) measurement of RSCC (reverse short-circuit current) in the absence of Na+ transport, (ii) inhibition of RSCC by acetazolamide, an inhibitor of carbonic anhydrase, and (iii) direct measurement of the pH change of the mucosal (urinary) fluid. As in the case of Na+ transport, spirolactone inhibited the action of aldosterone. Although not a unique model, the apparent paradoxical mimicry of aldosterone's stimulation of urinary acidification may be explained by a model which includes action of aldosterone and the inhibitors via their known effects on RNA and protein synthesis.

Aldosterone↗