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T Numazawa

Publications and source records attributed to T Numazawa.

13 recordsLinked to original sources
PubMed2000

Discovery of a muscarinic M3 receptor antagonist with high selectivity for M3 over M2 receptors among 2-[(1S,3S)-3-sulfonylaminocyclopentyl]phenylacetamide derivatives.

In the course of developing a metabolically stable M3 receptor antagonist from the prototype antagonist, J-104129 (1), introduction of certain substituents into the cyclopentane ring of 1 was found to be effective not only in improving metabolic stability but also in greatly enhancing the subtype selectivity. Among the cyclopentane analogues, sulfonamide derivatives (10f) and (10g) displayed 160- and 310-fold selectivity for M3 over M2 receptors, and both were significantly more selective than the prototype antagonist (120-fold). Subsequent derivatization of the sulfonamide series led to the highly selective M3 receptor antagonists (10h, 10i and 10j) with >490-fold selectivity for M3 over M2 receptors. Among them, p-nitrophenylsulfonamide (J-107320, 10h) exhibited 1100-fold selectivity for M3 receptors (Ki = 2.5 nM) over M2 receptors (Ki = 2800 nM) in the human muscarinic receptor binding assay using [3H]-NMS as a radio ligand.

Alkenes↗
PubMed1996

Mechanisms of desensitization of vasodilatation induced by platelet-activating factor in hypertensive rats.

We found that vasodilator effects of platelet-activating factor (PAF) on the mesenteric arterial bed of the rat were significantly attenuated in spontaneously hypertensive rats (SHR) and renal hypertensive rats (RHR). Perfusion of the mesentery with acetylcholine and PAF caused endothelium-dependent vasodilatation accompanied by an increase in cyclic GMP levels in the mesentery from normotensive Wistar Kyoto rats (WKY). Acetylcholine caused a significant increase in cyclic GMP levels in the effluent in both SHR and RHR, whereas PAF could not increase cyclic GMP levels in SHR and slightly increased cyclic GMP in RHR. Incubating the mesentery with PAF markedly inhibited the vasodilatation induced by PAF, but not acetylcholine or sodium nitroprusside. The cyclic GMP accumulation in the effluent was impaired in the mesenteric arterial bed pretreated with PAF and in that obtained from rats given islet-activating protein (IAP). The PAF-induced vasodilatation was completely reversed by the PAF receptor antagonist, CV-6209 (2-[N-acetyl-N-(2-methyl-3-octadecylcarbamoyl-oxypropoxycarbony l) aminomethyl]-1-ethylpyridinium chloride). These results suggest that (1) attenuated vasodilator effects of PAF and decreased cyclic GMP levels in the mesentery from SHR and RHR are due to desensitization but not to impairment of the endothelium; (2) GTP-binding protein, which is IAP-sensitive, may be involved in PAF-induced vasodilatation and cyclic GMP accumulation; (3) desensitization of the mesentery to PAF in SHR and RHR may be due to PAF receptor and GTP-binding protein uncoupling.

Acetylcholine↗
PubMed1996

Characteristics of vasodilatation induced by acetylcholine and platelet-activating factor in the rat mesenteric arterial bed.

We examined the nature of the endothelium-dependent vasodilator effects of acetylcholine and platelet-activating factor (PAF) on the perfused mesenteric arterial bed of the rat. Acetylcholine-induced concentration-dependent vasodilatation of the mesentery was not affected by pretreatment with 10(-4) M NG-monomethyl-L-arginine (L-NMMA), indomethacin, ouabain, or glibenclamide, whereas pretreatment with 10(-5) M oxyhemoglobin, 10(-5) M methylene blue, or 10 mM tetraethylammonium shifted the concentration-response curves to the right. PAF-induced concentration-dependent vasodilatation of the mesentery was inhibited by pretreatment with L-NMMA, oxyhemoglobin, or methylene blue, and slightly but significantly inhibited by tetraethylammonium, whereas indomethacin, glibenclamide, and ouabain had no inhibitory effects. PAF-induced vasodilatation of the mesentery was more sensitive to nitric oxide-cyclic GMP pathway inhibitors (a combined application of L-NMMA, oxyhemoglobin, and methylene blue) than was the vasodilatation induced by acetylcholine. Perfusion of the mesentery preparations with acetylcholine or PAF increased the levels of cyclic GMP in the effluent. These effects were completely inhibited by L-NMMA or oxyhemoglobin. These results suggest that the endothelium-dependent vasodilator effects of PAF are primarily mediated by endothelium-derived nitric oxide (NO) and those of acetylcholine are mediated by both NO and endothelium-derived hyperpolarizing factor (EDHF).

Acetylcholine↗
PubMed1994

Decrease in vasodilator effects of platelet-activating factor in resistance vessels of spontaneously hypertensive rats.

We studied the endothelium-dependent vasodilator effects of platelet-activating factor (PAF) on the mesenteric arterial bed of Wistar Kyoto rats (WKYs) and spontaneously hypertensive rats (SHRs) and found that vasodilation induced by PAF was significantly attenuated in SHRs. We discovered no significant differences in reactivity or sensitivity of the mesentery to acetylcholine between WKYs and SHRs. Our results strongly suggest that the attenuated vasodilator effects of PAF on the SHR mesenteric arterial bed may be due in part to desensitization rather than impairment of the endothelium.

Animals↗
PubMed1994

Vasodilator effects of clonidine on the mesenteric arterial beds in normotensive and spontaneously hypertensive rats.

Clonidine, an alpha 2-agonist, caused a concentration-dependent vasodilation of the mesenteric arterial beds in both normotensive and hypertensive rats. The clonidine-induced vasodilation was inhibited by NG-nitro-L-arginine, and the inhibition was reversed by L-arginine. The concentration-dependent vasodilation was not significantly different between normotensive and hypertensive rats. These results suggest that clonidine has an endothelium-dependent vasorelaxant action in the resistance artery such as rat mesenteric arterial bed, and the endothelium-dependent vasodilator effects of clonidine in the mesentery may be involved in the depressor effect of the drug.

Animals↗
PubMed1993

Biochemical consequences of biotin deficiency in osteogenic disorder shionogi rats.

The biological consequences of biotin deficiency in rats were investigated using osteogenic disorder Shionogi rats which have a hereditary defect in ascorbic acid synthesizing ability. Decrease of liver ascorbic acid content and fasting plasma glucose and an increase of plasma non-esterified fatty acid (NEFA) appeared in biotin deficient rats fed a diet containing 200 mg ascorbic acid per 100 g diet, compared with the pair fed control. On the other hand, in the case of rats fed a diet containing 500 mg ascorbic acid, although the clinical features of biotin deficiency developed, the ascorbic acid contents of liver and adrenal gland increased in comparison with those of AsA 200 mg groups, and the alterations of plasma levels of glucose and NEFA were improved partially in glucose and greatly in NEFA, respectively. This suggests that ascorbic acid may be consumed in the improvement of the metabolic impairments induced by biotin deficiency.

Animals↗
PubMed1991

Cis-acting elements in the pyruvate, orthophosphate dikinase gene from maize.

To investigate the mechanisms that control expression of the gene for pyruvate, orthophosphate dikinase (PPDK) in maize, the 5' flanking region of the gene was analyzed for interactions with nuclear extracts. Gel retardation assays showed that there are several sites in the promoter region which bind to protein factors. In this report we describe further study of one of these sites, designated the PPD-1 binding site. The nuclear binding factor, PPD-1, is restricted to nuclear extracts from green leaves where the PPDK gene is expressed. No binding of PPD-1 was detected in tissues such as roots or etiolated leaves where the gene is not expressed in vivo. Gel retardation assays using deletion fragments from the promoter region and synthetic oligonucleotides, as well as exonuclease III protection assays, revealed that the site of PPD-1 binding lies between positions -301 and -296. To identify the functional role of the interaction between PPD-1 and its binding site, a deletion series of the promoter region was joined to a reporter gene, beta-glucuronidase. These constructs were introduced into green leaves of maize by microprojectile bombardment. Expression of the reporter gene occurred if the PPD-1 binding site remained in the promoter region of the chimeric genes but deletion of the binding site caused a drastic reduction in expression levels. These data indicate that interaction between PPD-1 and its binding site is essential for active transcription of the PPDK gene.

Base Sequence↗
PubMed1991

Characteristics of two lines of guinea pigs (BHS and BHR) differing in bronchial sensitivity to acetylcholine and histamine exposure.

In a previous study, we reported that as model animals to be used in the study of bronchial asthma in humans, two lines of guinea pigs were developed by ourselves: bronchial hypersensitive line (BHS) and bronchial hyposensitive line (BHR) as a control. Studies on biological characteristics in guinea pigs of two lines were undertaken, and the following results were obtained. 1) Airway resistance of guinea pigs of the two lines to intravenously administered acetylcholine, histamine and leukotriene D4 was found to be different between BHS and BHR. Airway resistance of BHS to the chemicals was increased compared with those of BHR. 2) The number of muscarinic acetylcholine receptors in lung membrane preparation and its affinity increased significantly in BHS compared with those of BHR. In beta-adrenergic and histamine H1 receptors, there was observed no difference between BHS and BHR. 3) No difference in IgE antibody production to ovalbumin was observed between BHS and BHR. 4) When total leukocytes and differential leukocyte count (percentage, %) in peripheral blood of BHS and BHR were examined, relative percentage of lymphocytes and eosinophils was significantly higher in BHS than in BHR, while percentage of neutrophils was significantly lower in the former than in the latter.

Acetylcholine↗
PubMed1989

Aspartate aminotransferase from Panicum maximum Jacq. var. trichoglume Eyles, a C4 plant: purification, molecular properties, and preparation of antibody.

Extracts of the leaf tissue of Panicum maximum Jacq. var. trichoglume Eyles (a phosphoenolpyruvate carboxykinase type of C4 plant) were examined and at least two isoforms of aspartate aminotransferase (EC 2.6.1.1), with different electrophoretic mobilities, were detected. The predominant isoform was purified to homogeneity from mesophyll cells. The purification procedure included fractionation with ammonium sulfate followed by chromatography on diethylaminoethyl-cellulose, Sephacryl S-300, and hydroxyapatite. The purified enzyme had specific activities of 182 and 165 mumol/min/mg protein, measured in terms of the synthesis of oxaloacetate and aspartate, respectively, at pH 8.0. The enzyme, with an apparent molecular size of 100 kDa, appears to be a dimer of a single polypeptide with a molecular size of 42 kDa. Mono specific polyclonal antibodies were raised against the 42-kDa polypeptide. Only a single stained band was detected in extracts of whole leaves by immunoblot analysis with this antibody after two-dimensional polyacrylamide electrophoresis. Furthermore, no difference in mobility was observed between the enzymes extracted from mesophyll and bundle sheath cells on native polyacrylamide gels. These findings are discussed in relation to the other isoform in the leaves of this species.

Antibodies↗