Biomedical subjects
S Muto
Publications and source records attributed to S Muto.
Stereoselective action of (R*,R*)-(+/-)-methyl-4-[2-[2-hydroxy-2-(3-chlorophenyl)ethylamino] propyl]-phenoxyacetic acid (BRL37344) on beta-adrenoceptors and metabolic chiral inversion.
Stereoisomers of BRL37344 ((R*,R*)-(+/-)-methyl-4-[2-[2-hydroxy-2 -(3-chlorophenyl)ethylamino]propyl]-phenoxyacetic acid), a beta 3-adrenoceptor agonist, were synthesized and separated with good resolution by derivatization with 1-anthroyl cyanide prior to chiral HPLC. Agonist effects on rat right atria, guinea pig trachea, and rat brown adipocytes were due principally to the (RR) isomer, while other isomers (SS, RS, and SR) were much less potent or inactive. Since the racemate (RR +/- SS) was half as potent as the (RR) isomer in all specimens tested, the (SS) isomer does not appear to have antagonistic effects. When [14C](RR)BRL35135A ((R*,R*)-(+/-)-methyl-4-[2-[2-hydroxy-2-(3-chlorophenyl)ethylamino]propy l] -phenoxyacetate hydrobromide), the HBr salt of the methyl ester of BRL37344, was administered orally to male Wistar rats, both the (RR) and (SR) isomers of [14C]BRL37344 were detected in plasma, while only the (SS) isomer of [14C]BRL37344 was detected after [14C](SS)BRL35135A administration. These findings indicate that there is clear stereoselectivity in the effects of BRL37344 on beta-adrenoceptors, and that stereoselective chiral inversion from the RR isomer to the SR isomer occurs in rats.
Transcriptional activation of RACTK1 K+ channel gene by apical alkalization in renal cortical collecting duct cells.
We have previously demonstrated that RACTK1 cDNA encodes a pH sensitive K+ channel expressed in the apical side of renal collecting tubule cells. To determine whether extracellular pH induces the RACTK1 gene expression in the renal cortical collecting duct (CCD) cells, we measured mRNA of the RACTK1 using cultured rabbit CCD cells. Alkalization of incubation medium activated the transcription of the RACKTK1 gene in a time- and dose-dependent manner after 1 h, and reached a maximal level after 12 h. To examine whether the stimulation of mRNA by alkalization of body fluid occurs also in vivo, mRNA levels were measured in mice loaded with acid or alkali. The RACTK1 mRNA was increased in association with the rise in urinary pH. To examine side face of the effect of pH on stimulation of mRNA, we observed the effect of pH in the apical or the basolateral side in the preparation where CCD cells were cultured on filter membrane supports. Alkalization of the apical side but not of the basolateral side, was shown to be a determinant in inducting the RACTK1 mRNA. These findings suggest that, in addition to rapid direct regulation of RACTK1 K+ channel conductance by intracellular pH, this channel is also regulated by the changes in luminal pH through synthesis of channel protein by transcriptional activation.
Magnetic properties of Pm in NdNi.
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Dipyridamole enhances interleukin-1beta-stimulated nitric oxide production by cultured rat vascular smooth muscle cells.
We examined whether dipyridamole affected interleukin-1beta-stimulated nitric oxide (NO) production by cultured rat vascular smooth muscle cells. Interleukin-1beta stimulated the production of nitrite and nitrate, stable metabolites of NO, in a dose- and time-dependent manner in vascular smooth muscle cells. Dipyridamole (1-100 mu M) enhanced interleukin-1beta-induced nitrite production in a dose- and time-dependent manner. The mRNA expression of inducible NO synthase was up-regulated by dipyridamole (0.3-10 mu M) treatment in a dose-dependent manner. Both 8-bromo-guanosine 3',5'-cyclic monophosphate (8-bromo-cGMP) and dibutyryl adenosine 3',5'-cyclic monophosphate (db-cAMP) enhanced the nitrite production in the presence of interleukin-1beta. Dipyridamole up-regulated the effect of both 8-bromo-cGMP and db-cAMP on the interleukin-1beta-induced nitrite production. Dipyridamole increased the intracellular cAMP content in the presence of interleukin-1beta (10 ng/ml), but did not affect the intracellular cGMP content. 8R*,9S*,11S*-(-)-9-hydroxy-9-n-hexyloxy-8-methyl-2,3,9,10- tetrahydro-8,11-epoxy-1H,8H,11H-2,7b,11a-triazadibenzo-(a,g)-cy cloocta ++-(c,d,e)-trinden-1-one (KT 5720), a selective inhibitor of cAMP-dependent protein kinase, abolished the enhancement of interleukin-1beta-induced nitrite production by dipyridamole, whereas 8R*,9S*,11S*-(-)-9-methoxy-carbamyl-8-methyl-2,3,9,10-tetrahydro-8,11-ep oxy-1H,8H,11H-2,7b,11a-trizadibenzo-(a,g)-cyclo-octa-(c,d,e)-tr inden-1-one (KT 5823), an inhibitor of cGMP-dependent protein kinase, did not attenuate the enhancement. Furthermore, Rolipram and 4-(3-butoxy-4-methoxybenzyl)-2-imidazolidinone (Ro-20-1724), cAMP-specific phosphodiesterase type IV inhibitors, augmented the interleukin-1beta-induced nitrite production. We concluded that dipyridamole enhanced the interleukin-1beta-induced NO production via an increase in intracellular cAMP content in cultured rat vascular smooth muscle cells.
Measurements for spin inversion and noninversion in successive decays via nuclear magnetic resonance on oriented nuclei.
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Measurement of the nuclear magnetic moments of 57Ni and 59Fe.
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Differential regulation of Na+-K+-ATPase gene expression by corticosteriods in vascular smooth muscle cells.
To determine whether gluco- and mineralocorticoids have specific actions on Na+-K+-ATPase gene expression in vascular tissue, we used Northern blot analysis to compare the effects of dexamethasone (Dex) and aldosterone (Aldo) on Na+-K+-ATPase alpha1 and beta1-subunit mRNA expression in cultured vascular smooth muscle cells from rat aortae. Dex at 10(-6)M increased alpha1 -mRNA level 2.5-fold at 24 h and beta1-mRNA level 9.9-fold at 12 h. Aldo at 10(-6)M increased alpha1-mRNA 2.7-fold at 48 h and beta1-mRNA level 10.9-fold at 6 h. The half-maximal stimulation of both alpha1 and beta1-mRNA levels occurred at a concentration of 5-7 X 10(-9)M Dex, whereas it occurred at a concentration of 2-3 X 10(-9)M Aldo. The glucocorticoid receptor antagonist RU-38486 inhibited both Dex- and Aldo-mediated induction of beta1-mRNA. The mineralocorticoid receptor antagonist spironolactone inhibited Aldo-mediated induction of beta1-mRNA, whereas it had no effect on Dex-mediated induction of beta1-mRNA. Removal of Na+ from the extracellular medium (isosmotic replacement with choline) caused no effect on Dex-mediated induction beta1-mRNA, whereas it inhibited Aldo-mediated induction of beta1-mRNA. Addition of a specific inhibitor of the Na+/H+ exchange, ethylisopropylamiloride, had no effect on Dex-mediated induction of beta1-mRNA, whereas it resulted in a significant inhibition of Aldo-mediated induction of beta1-mRNA. We conclude that 1) both Dex and Aldo induce Na+-K+-ATPase alpha1- and beta1-mRNA expression in a time- and dose-dependent manner; 2) Dex-mediated induction of beta1-mRNA occurs only through glucocorticoid receptors, whereas Aldo-mediated induction of beta1-mRNA occurs through both gluco- and mineralocorticoid receptors; and 3) Dex-mediated induction of beta1-mRNA occurs through Na+-independent mechanisms, whereas Aldo-mediated induction of beta1-mRNA, at least in part, occurs through Na+-dependent mechanisms, including stimulation of the Na+/H+ exchange.
Hyperosmolality stimulates Na-K-ATPase gene expression in inner medullary collecting duct cells.
Primary cultures of inner medullary collecting duct (IMCD) cells of rats were incubated in hyperosmotic media to determine the effects on Na-K-ATPase alpha 1- and beta 1-subunit mRNA expression. Osmolality of the incubation media was raised from 300 up to 500 mosmol/kgH2O by adding NaCl, mannitol, raffinose, or urea. Hyperosmotic media supplemented with NaCl, mannitol, or raffinose caused two- to fourfold increases in the alpha 1-subunit mRNA accumulation and five- to eightfold increases in the beta 1-subunit mRNA accumulation, with peak elevations of both subunits at 12 h after addition. In sharp contrast, hyperosmolar urea medium had no effect at any time. When NaCl or mannitol was added to the media in amounts ranging from 300 to 600 mosmol/kgH2O, the maximal effects on both alpha 1- and beta 1-subunit mRNA accumulation occurred at 500 mosmol/kgH2O. In urea-supplemented medium, however, there was no significant change at any level of osmolality. The upregulation of alpha 1- and beta 1-subunit mRNA induced by hyperosmotic mannitol- or raffinose-supplemented media was markedly inhibited by removal of Na from the culture medium. Furthermore, pretreatment with a protein synthesis inhibitor cycloheximide partially inhibited the upregulation of alpha 1- and beta 1-subunit mRNA in IMCD cells exposed to hyperosmotic media treated with NaCl or mannitol. When IMCD cells were incubated with hyperosmotic media (500 mosmol/kgH2O) supplemented with NaCl or mannitol for 24 h, Na-K-ATPase activity increased by 78.6 and 82.8%, respectively. In contrast, hyperosmolar urea medium had no significant effect on Na-K-ATPase activity. These results demonstrate that 1) hyperosmolality induced by the poorly permeating solutes (NaCl, mannitol, and raffinose) but not the rapidly permeating solute (urea) stimulates both alpha 1- and beta 1-subunit mRNA accumulations in IMCD cells in a time- and an osmolality-dependent manner, 2) the hyperosmolality-induced upregulation of alpha 1- and beta 1-subunit mRNA leads to an increase in Na- K -ATPase activity; and 3) the above upregulation of alpha1- and beta 1-subunit mRNA in response to hyperosmotic media requires, at least in part, the presence of Na in the extracellular medium and the de novo synthesis of intermediate proteins.
Nitric oxide and immune complexes are involved in the induction of a novel luminol chemiluminescence in cytotoxic macrophages.
A chemiluminescence (CL) was observed immediately after the addition of luminol to thioglycollate-elicited ICR mouse peritoneal macrophages (M phi) that had been incubated overnight with recombinant murine interferon-gamma (IFN-gamma) and lipopolysaccharides (LPS). The intensity of this CL was closely correlated with the cytotoxic activity of M phi. NG-monomethyl-L-arginine (L-NMMA), an inhibitor of nitric oxide (NO) synthase, inhibited the induction of this CL, and L-arginine restored the L-NMMA-induced inhibition. These results suggest that NO is directly involved in the induction of CL. However, we found that immune complexes are required for the induction of CL as well as NO.
Mechanisms of hyperkalemia caused by nafamostat mesilate.
1. Nafamostat mesilate (NM) is a novel serine-protease inhibitor used for the treatment of acute pancreatitis and disseminated intravascular coagulation. Recently, NM has been reported to cause hyperkalemia due to reduced urinary excretion of potassium (K). 2. This review briefly summarizes the roles of the cortical collecting duct (CCD) in the renal K excretion. 3. In vitro microperfusion technique was applied to examine whether NM, and its two metabolites, p-guanidinobenzoic acid (PGBA) and 6-amidino-2-naphthol, directly act on the CCD. 4. It was demonstrated that these compounds act mainly on the apical membrane of the collecting duct cell in the CCD and inhibit the amiloride-sensitive sodium (Na) conductance, resulting in an inhibition of K secretion. PGBA had the most potent action. 5. This direct action of these two metabolites, rather than NM, could contribute to the NM-induced hyperkalemia.
Effects of glucocorticoid and mineralocorticoid on potassium transport in the rat medullary thick ascending limb of Henle's loop.
Potassium transport in the thick ascending limb is conducted by heterogeneous cells to opposite directions. Effects of glucocorticoids and mineralocorticoids on potassium transport of the rat medullary thick ascending limb (MTAL) were examined by in vitro microperfusion technique by measuring net potassium flux (JK), and apparent potassium conductances in the apical and basolateral membranes of two cell populations. Segments of MTAL were obtained from four groups of rats: sham operated control rats, adrenalectomized rats (ADX), adrenalectomized rats treated with dexamethasone (DEX), and adrenalectomized rats treated with aldosterone (ALDO). Fractional urinary potassium excretion was reduced by ADX and partially recovered by either DEX or ALDO. JK of the isolated perfused MTAL was markedly decreased by ADX from 5.13 to 1.94 pmol/min/mm. It was partially recovered by DEX (3.36 pmol/min/mm), but not by ALDO (2.07 pmol/min/mm). Random impalement of MTAL cells with a microelectrode in the control group revealed two cell populations; 76% was high basolateral conductance cell (HBC) and 24% was low basolateral conductance cell (LBC). In the ADX group, the basolateral potassium conductance of the HBC cell was markedly reduced, whereas the apical membrane potassium conductance of the HBC cell was increased. These changes were recovered in the DEX group, but not in the ALDO group. Potassium conductances of the apical and basolateral membranes in the LBC were unchanged.(ABSTRACT TRUNCATED AT 250 WORDS)
Action of aldosterone on renal collecting tubule cells.
Aldosterone is a type of steroid hormone that acts primarily in renal collecting ducts to stimulate reabsorption of Na+ as well as secretion of K+ and H+. It binds with intracellular receptors in the nucleus that stimulate the expression of several genes. Transcription and subsequent translation result in the production of new proteins that modulate the activity of ionic transport systems located in the apical and basolateral membranes of the target epithelial cells. This review focuses on the cellular mechanisms of aldosterone action on Na+, K+, and H+ transport in mammalian renal collecting ducts. Although the cellular actions of aldosterone conform to the general mode of steroid hormone action, numerous questions relating to each step in the process remain unanswered.
Specific increase in interleukin-8 concentrations in dialysis fluid of patients with peritonitis receiving continuous ambulatory peritoneal dialysis.
AIMS: To evaluate the influence of interleukin-8 (IL-8) and other inflammatory cytokines (IL-6, IL-1 beta and tumour necrosis factor alpha (TNF alpha)) on the occurrence of peritonitis in patients receiving continuous ambulatory peritoneal dialysis (CAPD). METHODS: The study population comprised 12 patients with peritonitis, 33 without peritonitis, all undergoing CAPD, and five patients undergoing peritoneal catheter implantation. Cytokine concentrations in dialysis fluid were determined by immunoassay and their values compared. RESULTS: Concentrations of both IL-8 (median 147 pg/ml, range 20-2273 pg/ml; n = 12) and IL-6 (median 1120 pg/ml, range 96-10,600 pg/ml) were substantially elevated, while the IL-1 beta concentration was lower and TNF alpha was not detectable in patients at diagnosis. The IL-6 concentration was also elevated in patients undergoing catheter implantation as well as in those with peritonitis. The IL-8 concentration, however, was elevated only upon infection. Intraperitoneal production of IL-8 was evident on determination of paired serum and dialysis fluid cytokine concentrations, and immunostaining of peritoneal cells with monoclonal anti-IL-8 antibody. CONCLUSIONS: These results suggest that determination of the IL-8 concentration in dialysis fluid maybe useful as a specific marker for following patients with peritonitis receiving CAPD.
Potassium secretion is inhibited by metabolic acidosis in rabbit cortical collecting ducts in vitro.
The role of metabolic acidosis in the regulation of transepithelial potassium transport was examined in rabbit cortical collecting ducts (CCD) using in vitro isolated tubular microperfusion and conventional microelectrode techniques. Basolateral metabolic acidosis, created by reduction of bicarbonate concentration from 25 to 5 meq/l, pH 7.40 to 6.80, depolarized the transepithelial voltage significantly (-6.5 +/- 1.0 to -2.7 +/- 1.3 mV). Basolateral acidosis also suppressed net potassium secretion (-14.3 +/- 2.1 to -9.0 +/- 1.7 pmol.min-1.mm-1). Electrophysiological study in CCD cells demonstrated that basolateral metabolic acidosis depolarized transepithelial voltage and apical and basolateral membrane voltage with an increase of transepithelial and fractional apical resistance. Basolateral acidosis did not affect the 22Na efflux nor 86Rb efflux. The inhibitory action of basolateral acidosis on net potassium secretion remained in the presence of luminal barium and in the absence of bicarbonate. Ouabain could not abolish the effect of basolateral acidosis on transepithelial voltage completely. These data lead us to conclude that basolateral acidosis affects multiple transport pathways, and it inhibits mainly apical barium-sensitive potassium transport. Additionally, it inhibits apical sodium conductance, barium-insensitive potassium transport, and stimulates a ouabain-insensitive electrogenic transport pathway to some degree.
Gene transfer into circulating primordial germ cells of quail embryos.
During early stages in avian embryogenesis primordial germ cells (PGCs) show a unique migration pathway toward the gonadal anlage through the circulation. In the present study, liposomes consisting of plasmid DNA (pMiwZ; containing lacZ as a reporter) and Lipofectin were injected into the marginal veins of quail embryos during the stages PGCs were circulating in the blood vessels. The lacZ expression was then histochemically detected in the gonads at later embryonic stages, indicating the expression of the injected DNA in PGCs.
Effect of growth hormone on wound healing in protein-malnourished rats treated with corticosteroids.
This study was designed to evaluate the efficacy of human growth hormone (GH) in improving the tensile strength of wounds weakened by chronic protein malnourishment and corticosteroid (CS) administration. Eighty-six female Sprague-Dawley rats, weighing 80 to 100 g, were divided into five groups. Group 1 (control) received 23.4% protein chow for 8 weeks before surgery. Groups 2, 3, 4, and 5 received nonprotein chow on alternate days for the same duration. Groups 3 and 5 received prednisolone (2 mg/kg/d intramuscularly) for 3 weeks preoperatively and for 5 days postoperatively. Groups 4 and 5 were given GH (somatotropin, 1 IU/d) for 5 days postoperatively. All the animals underwent a precise 4-cm midline celiotomy. Wound testing was performed on the sixth postoperative day, after removal of the sutures. The bursting strength (BS, mean +/- SD) for group 1 was 145 +/- 16 mm Hg. The BS for groups 4 (137 +/- 13 mm Hg) and 5 (134 +/- 7 mm Hg) were significantly stronger than those for groups 2 (115 +/- 15 mm Hg) and 3 (91 +/- 16 mm Hg). The authors conclude that postoperative systemic GH restored the wound BS in protein-malnourished animals treated with CS, to the level of the normally nourished controls.
Mechanisms of the hyperkalaemia caused by nafamostat mesilate: effects of its two metabolites on Na+ and K+ transport properties in the rabbit cortical collecting duct.
1. The present experiments were undertaken to determine the mechanism(s) of hyperkalaemia caused by nafamostat mesilate (NM), a serine-protease inhibitor. 2. We investigated the effects of luminal addition of two metabolites of NM, p-guanidinobenzoic acid (PGBA) and 6-amidino-2-naphthol (AN), on Na+ and K+ transport properties of the collecting duct (CD) cell in the isolated perfused cortical collecting duct (CCD) from rabbit kidneys, because these metabolites, but not NM, were mainly excreted into the urine. 3. Addition of PGBA at 10(-5) and 10(-4) M in the lumen resulted in a hyperpolarization of VA in parallel with increases in transepithelial resistance (RT) and fractional apical membrane resistance (fRA). PGBA added to the luminal perfusate at 10(-5) and 10(-4) M changed VA, RT and fRA in a dose-dependent manner. These effects were completely inhibited by pretreatment with luminal amiloride (50 microM). PGBA at 10(-6) M in the lumen had no effect on the electrical parameters. 4. Luminal addition of AN at 10(-4) M also caused the apical membrane to hyperpolarize in parallel with increases in RT and fRA. These effects were also completely inhibited by pretreatment with luminal amiloride (50 microM). AN at 10(-5) M in the lumen had no effect on the electrical parameters. 5. We conclude that two metabolites of NM, PGBA and AN, act on the apical membrane of the CD cell and inhibit the amiloride-sensitive Na+ conductance, resulting in an inhibition of K+ secretion. This direct action of these metabolites, rather than NM, on the CCD might contribute to the NM-induced hyperkalaemia.