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Biomedical subjects

T Asano

Publications and source records attributed to T Asano.

At least 919 records · Page 51Linked to original sources
PubMed1977

Cyclic 3':5'-nucleotide phosphodiesterase determined in various human tissues by DEAE-cellulose chromatography.

Tissue extracts from human heart, lung, liver, kidney, skeletal muscle and cerebrum displayed at least 3 distinct cyclic 3':5'-nucleotide phosphodieterase (EC 3.1.4.17) activity peaks (FI, FII, FIII) on DEAE-cellulose chromatography and various properties of these forms were compared in each tissue. FI eluted at about 0.08 M sodium acetate, hydrolyzed cyclic GMP more rapidly than it did cyclic AMP, and cyclic GMP hydrolysis by FI in most tissues was enhanced by a protein activator in the presence of CaCl2. As only high concentrations of cyclic AMP inhibited cyclic GMP hydrolytic activity of FI, the enzyme probably has a low affinity for cyclic AMP. FII eluted at about 0.2 M sodium acetate, hydrolyzed both nucleotides at equal rates, and substrate affinities were relatively low. Cyclic GMP hydrolysis by FII was also stimulated by addition of a protein activator in the presence of CaCl2 and cyclic AMP hydrolysis in this fraction was accelerated by a micromolar fraction of cyclic GMP. FII eluted at about 0.35 M hydrolyzed cyclic AMP preferentially and was insensitive to protein activator. These two cyclic nucleotides act as mutual inhibitors of the hydrolysis in this fraction. Ratio of the cyclic GMP to cyclic AMP hydrolysis was in the order FI, FII, FIII. Four activity peaks were eluted from the cerebral extract and enzymes from this tissue exhibited much the same properties as observed in the other tissues examined herein.

3',5'-Cyclic-AMP Phosphodiesterases↗
PubMed1977

Resistance of germfree rats to indomethacin-induced intestinal lesions.

Indomethacin given orally to conventional rats produced in three days a syndrome, often fatal, of intestinal lesions characterized by multiple ulcers and peritonitis. Male germfree rats were found to be resistant to this effect of indomethacin, while female germfree rats developed very mild lesions. Germfree rats became sensitive again to such lesions when monocontaminated with E. coli. In such animals, however, the lesions were less severe than in conventional animals, presumably because more than one microorganism is necessary for the full syndrome to develop. These results suggest that microorganisms are necessary for the development of indomethacin-induced intestinal lesions. Secondary bile acids, absent in germfree animals, may also be necessary. The prostaglandin deficiency caused by indomethacin appears to weaken the resistance of the intestinal mucosa to microorganisms and/or their toxins. The latter may then penetrate the mucosa, damage the cells and produce ulcers and perforations. Since several prostaglandins also protect against indomethacin-induced lesions, the hypothesis is advanced that certain prostaglandins may protect the mucosa ("cytoprotection") by preventing the spread of microorganisms and/or their toxin through the intestinal wall.

Animals↗
PubMed1977

Stimulation of human platelet guanylate cyclase by unsaturated fatty acid peroxides.

Guanylate cyclase [GTP pyrophosphate-lyase (cyclizing), EC 4.6.1.2] activity of human platelet homogenates was stimulated by the addition of phospholipase A2 or unsaturated fatty acids such as oleic, vaccenic, linoleic, linolenic, eicosenoic, eicosadienoic, and arachidonic acids. The addition of lipoxidase potentiated the fatty acid-induced stimulation of guanylate cyclase purified by DEAE-cellulose column chromatography. The extent of the stimulation was dependent on the concentration of the oxidized form of these fatty acids (peroxides). Saturated fatty acids such as stearic and arachidic acids had no effect on the guanylate cyclase activity in the presence or absence of lipoxidase, indicating that human plateletguanylate cyclase is stimulated by unsaturated fatty acid peroxides rather than by fatty acids. Hemoglobin prevented the enzyme stimulation produced by low concentrations of fatty acid peroxides, but enhanced stimulation of the enzyme activity with high concentrations of fatty acid peroxides. 2-Mercaptoethanol, dithiothreitol, and N-ethylmaleimide inhibited the guanylate cyclase activities both in the presence and absence of unsaturated fatty acidperoxide. The stimulation of guanylate cyclase activity by unsaturated fatty acid peroxidesis attributed to oxidation of sulfhydryl residues of the enzyme protein.

Arachidonic Acids↗
PubMed1977

Xylitol, a partial homologue ot alpha-D-glucopyranose: potent stimulator of insulin release in dogs.

A direct comparison was made in healthy female dogs of the potency of xylitol and mutarotated glucose as stimulators of insulin release (both first and second phases), and also of their urinary excretion and arterio-venous difference across a hind-limb. Xylitol or glucose was given by constant infusion into a systemic vein for 50 minutes in paired experiments. Three dose levels were used in 12 dogs. Plasma insulin levels during xylitol infusion were as high as or higher than those during glucose infusion, with increases in arterial xylitol levels equal to or less than those in glucose. More xylitol was lost in the urine, but the overall uptake of xylitol was at least 70% also the arteriovenous difference for xylitol across the hind-limb was greater than for glucose, when each was divided by the arterial sugar concentration. It is suggested that both the striking potency of xylitol in stimulating both phases of insulin release, and its rapid uptake by tissues, may result from its homology with alpha-D-glucopyranose, the alpha-anomer of glucose. The concept of a glucoreceptor on the surface of the pancreatic beta-cell, with stereospecificity for alpha-D-glucopyranose or closely similar molecules, is supported by our results.

Animals↗
PubMed1977

Adaptive properties of the b-wave and the PIII in the perfused isolated carp retina.

1. Perfusion of isolated carp retina with a pure oxygen saturated standard solution used in this experiment permitted several continuous hours of ERG recordings showing a distinctive b-wave as well as the PIII isolated by sodium aspartate. 2. The dark-adapted threshold for the PIII required an absorption of 5 to 6 quanta per rod. The 'rod-cone break' in the increment threshold curve for the b-wave was induced at a background intensity of about 2.0 X 10(-2) muW/cm2. 3. Both the b-wave and the PIII showed rapid and complete recovery of sensitivity after exposure to dim adapting lights. After bright light adaptation, the sensitivities went through a short 'fast phase' followed by a prolonged 'slow phase' of recovery. The rate of recovery during the fast phase was significantly faster for the PIII than for the b-wave. Neither a lowered temperature (9 degrees C) nor the application of ouabain (10(-6) M) influenced the fast phase. This seems to prove that the visual pigment, photoproducts and the active transport hardly participate in its mechanism, if at all. 4. The prolonged slow phase that followed the fast phase was distinguished by the recovery of the rod activity, even in isolated retinas.

Adaptation, Ocular↗
PubMed1977

Pathogenetic role of no-reflow phenomenon in experimental subarachnoid hemorrhage in dogs.

The real pathogenetic role of no-reflow phenomenon in clinical situations such as the acute stage of subarachnoid hemorrhage (SAH) is not yet known. To study this problem, we carried out the following experiment in dogs: SAH was induced by withdrawing a needle previously inserted into the internal carotid artery through a small craniectomy in the lateral base of the skull. Complete dural repair and cranioplasty was done to avoid cerebrospinal fluid leakage. Cortical cerebral blood flow (CBF) changes, measured by a double-needle type thermocouple, intracranial pressure (ICP), electroencephalogram (EEG), and sensory evoked response were monitored under controlled ventilation of 3 hours after SAH. At the end of the experiment, the brain was perfused with carbon black solution at a pressure of 120mm Hg. The 32 episodes of SAH thus induced yielded two basic patterns of ICP changes which simulated those previously reported with human SAH. In the first pattern, reactive hyperemia was always observed, followed by complete or incomplete recovery of cerebral function. Perfusion defects were frequently seen in the thalamus, basal ganglia, and parietooccipital cortex symmetrically. In the second pattern, prolonged elevation of ICP resulted in failure of recovery of both CBF and EEG. Carbon black filled only the pial arteries and the rest of the brain was totally unperfused. From the results, the pathogenetic role of the no-flow phenomenon in the acute stage of SAH as influencing the prognosis is strongly suspected.

Acute Disease↗
PubMed1976

Platelet cyclic 3':5'-nucleotide phosphodiesterase released by thrombin and calcium ionophore.

Contact of rat platelets with thrombin or the divalent cation ionophore A-23187, in the presence of extracellular calcium, resulted in the secretion of adenosine 3':5'-monophosphate (cyclic AMP) and guanosine 3':5'-monophosphate (cyclic GMP) phosphodiesterases. Significant association of calcium with platelets occurred during platelet surface contact with thrombin. Thrombin concentration to induce association of calcium virtually agreed with that to release the enzyme. The finding that A-23187 (5 to 20 muM) also provoked a rapid and marked association of extracellular calcium with platelets suggests that calcium mobilization into the intracellular environment may account, at least in part, for this association between platelet and calcium. Two different phosphodiesterases, a relatively specific cyclic AMP and a relatively specific cyclic GMP phosphodiesterase were secreted from platelets into the plasma in soluble form. The amounts of the phosphodiesterases secreted were dose- or time-dependent on thrombin (0.1 to 2 units) or A-23187 (5 to 20 muM) within 30 min. The enzyme release by thrombin was completely inhibited by heparin but the release by A-23187 was not. The two phosphodiesterases secreted seemed to correspond to the two enzymes isolated from platelet homogenates in many respects. Rat platelets contained, at least, three cyclic 3':5'-nucleotide phosphodiesterases, namely, two relatively specific cyclic AMP phoshodiesterases and a relatively specific cyclic GMP phosphodiesterase which were clearly separated from each other by Sepharose 6B or DEAE-cellulose column chromatography or sucrose gradient centrifugation. The two platelet cyclic AMP phosphodiesterase (Mr = 180,000 and 280,000) had similar apparent Km values of 0.69 and 0.75 muM with different sedimentation coefficient values of 4.9 S and 7.1 S, respectively. They did not hydrolyze cyclic GMP significantly. A cyclic GMP phosphodiesterase (Mr - 260,000) exhibited abnormal kinetics for cyclic GMP with an apparent Km value of 1.5 muM and normal kinetics for cyclic AMP with a Km of 300 muM. The properties of a platelet cyclic AMP phosphodiesterase (Mr = 180,000) and a platelet cyclic GMP phosphodiesterase were found to agree with those of the two phosphodiesterases released from platelets by thrombin or A-23187. Depletion of extracellular calcium by an addition of citrate, EDTA, or ethylene glycol bis(beta-aminoethyl ether)-N,N'-tetraacetic acid (EGTA) to the blood or platelet suspension resulted in a loss of the activity of the smaller form of platelet cyclic AMP phosphodiesterase (Mr = 180,000) and addition of calcium restored the activity of this cyclic AMP phosphodiesterase. Thus, calcium seemed to be involved in the mechanism of an occurrence of this smaller form of cyclic AMP phosphodiesterase as well as the secretion of this enzyme. Contact of human platelets with thrombin also resulted in the secretion of cyclic nucleotide phosphodiesterase which was dependent on the concentration of calcium. No species difference was observed in this respect.

3',5'-Cyclic-AMP Phosphodiesterases↗
PubMed1976

Human blood platelet 3': 5'-cyclic nucleotide phosphodiesterase. Isolation of low-Km and high-Km phosphodiesterase.

Human blood platelet contained at least three kinetically distinct forms of 3': 5'-cyclic nucleotide phosphodiesterase (3': 5'-cyclic-AMP 5'-nucleotidohydrolase, EC 3.1.4.17) (F I, F II, and F III) which were clearly separated by DEAE-cellulose column chromatography. Although a few properties of the platelet phosphodiesterases such as their substrate affinities and DEAE-cellulose profile resembled somewhat those of the three 3': 5'-cyclic nucleotide phosphodiesterase in rat liver reported by Russell et al. [10], there were pronounced differences in some properties between the platelet and the liver enzymes: (1) the platelet enzymes hydrolyzed both cyclic nucleotides and lacked a highly specific cyclic guanosine 3': 5'-monophosphate (cyclic GMP) phosphodiesterase and (2) kinetic data of the platelet enzymes indicated that cyclic adenosine 3': 5'-monophosphate (cyclic AMP) and cyclic GMP interact with a single catalytic site on the enzyme. F I was a cyclic nucleotide phosphodiesterase with a high Km for cyclic AMP and a negatively cooperative low Km for cyclic GMP. F II hydrolyzed cyclic AMP and cyclic GMP about equally with a high Km for both substrates. F III was low Km phosphodiesterase which hydrolyzed cyclic AMP faster than cyclic GMP. Each cyclic nucleotide acted as a competitive inhibitor of the hydrolysis of the other nucleotide by these three fractions with Ki values similar to the Km values for each nucleotide suggesting that the hydrolysis of both cyclic AMP and cyclic GMP was catalyzed by a single catalytic site on the enzyme. However, cyclic GMP at low concentration (below 10 muM) was an activator of cyclic AMP hydrolysis by F I. Papaverine and EG 626 acted as competitive inhibitors of each fraction with virtually the same Ki value in both assays using either cyclic AMP or cyclic GMP as the substrate. The ratio of cyclic AMP hydrolysis to cyclic GMP hydrolysis by each fraction did not vary significantly after freezing/thawing or heat treatment. These facts also suggest that both nucleotides were hydrolyzed by the same catalytic site on the enzyme. The differences in apparent Ki values for inhibitors such as cyclic nucleotides, papaverine and EG 626 would indicate that three enzymes were different from each other. Centrifugation in a continuous sucrose gradient revealed sedimentation coefficients F I and II had 8.9 S and F III 4.6 S. The molecular weight of these forms, determined by gel filtration on a Sepharose 6B column, were approx. 240 000 (F I and II) and 180 000 (F III). F III was purified extensively (70-fold) from homogenate, with a recovery of approximately 7%.

3',5'-Cyclic-AMP Phosphodiesterases↗
PubMed1976

The size pH, and redox potential of the cecum in mice associated with various microbial floras.

Cecal size and in situ redox potential and pH of cecal contents were determined in conventionally reared mice and mice reared under a variety of gnotobiotic conditions: germfree, monoassociated with a cecal Clostridium sp., hexaflora-associated and thermoduric polyflora-associated. The mean Eh was approximately +200 mV in germfree and -200 mV in conventional mice. The Eh was close to zero in the monoassociated mice, thus occupying a position intermediate between the germfree and conventional mice. The potentials observed in the hexaflora and the thermoduric flora groups were indistinguishable from those of conventional animals. The degree of normalization was more advanced with respect to the redox potential than to the cecal size in the various gnotobiotic groups. In the thermoduric polyflora-associated group, normalization was observed in both cecal size and redox potential. This demonstrates that normalization can be accomplished with a relatively simplified microflora, at least with regard to the parameters studied.

Animals↗
PubMed1975

3':5'-cyclic-nucleotide phosphodiesterases of mammalian sera.

Cyclic AMP and cyclic GMP phosphodiesterases (3':5'-cyclic-nucleotide 5'-nucleotidohydrolase EC 3.1.4.17) were found in the sera of human, dog, rabbit and rat. The formed elements of blood were not present in sera and thus not the source of the phosphodiesterase. More rapid hydrolysis of cyclic GMP than cyclic AMP is observed in the sera of these four species when 0.4 muM of cyclic AMP or cyclic GMP is used as the substrate. Protein activator of the phosphodiesterase is not detectable in the sera of these four species. Serum cyclic AMP and cyclic GMP phosphodiesterase activities are not stimulated by protein activator prepared from bovine brain. The serum phosphodiesterases of these four species are purified through Sepharose 6B column chromatography. Cyclic AMP phosphodiesterase are found in a broad area corresponding to molecular weights ranging from approximately 150 000 to 340 000 with 2 to 3 peaks in all animals tested. Cyclic GMP hosphodiesterase is found in a single area corresponding to molecular weights of 230 000 (rabbit and rat) and 270 000 (human and dog). Serum cyclic AMP and cyclic GMP hosphodiesterase activities of these four species have pH optimum of 7.5-8.5. Optimal concentration of Mg-2+ is about 5 mM for cyclic GMP phosphodiesterase activities of these four species as well as for cyclic AMP phosphodiesterase activities except rabbit. Rabbit serum cyclic AMP phosphodiesterase requires higher concentration of Mg-2+ (50 mM) for its optimal activity. The double reciprocal plots are non-linear for cyclic AMP phosphodiesterases of all animals and cyclic GMP phosphodiesterases of human, dog and rat. Rabbit cyclic GMP phosphodiesterase exhibits a linear reciprocal plot. Cyclic GMP is inhibitor of serum cyclic AM phosphodiesterase. Rabbit enzyme was most effectively inhibited by cyclic GMP.

3',5'-Cyclic-AMP Phosphodiesterases↗