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Biomedical subjects

T Oshima

Publications and source records attributed to T Oshima.

At least 379 records · Page 21Linked to original sources

Molecular cloning of genes encoding major two subunits of a eubacterial V-type ATPase from Thermus thermophilus.

The atpAB genes which encode the alpha and beta subunits of membrane ATPase from a thermophilic eubacterium, Thermus thermophilus HB8, were cloned. The deduced amino-acid sequences of the alpha subunit (583 amino acids) and the beta subunit (478 amino acids) are only moderately similar to the alpha beta subunits of the F0F1-ATPases, while they are highly similar to the major two subunits of the V-type ATPases, a family of ATPases which have been so far found in eukaryotic endomembrane vacuolar vesicles and archaebacterial plasma membranes. Thus, T. thermophilus ATPase belongs to the V-type ATPase family, even though this bacterium is a eubacterium. The hypothesis that the differentiation of an ancestral ATPase into V-type and F0F1-ATPase occurred after the evolution of a primordial cell into archaebacteria and eubacteria should be modified accordingly.

Adenosine Triphosphatases↗

cAMP-dependent induction of fatty acid cyclooxygenase mRNA in mouse osteoblastic cells (MC3T3-E1).

In an osteoblastic cell line, MC3T3-E1, cloned from mouse calvaria, epinephrine stimulated the production of prostaglandin E2 as an essentially sole arachidonate metabolite (Kusaka, M., Oshima, T., Yokota, K., Yamamoto, S., and Kumegawa, M. (1988) Biochim. Biophys. Acta. 972, 339-346). Western and Northern blot analyses showed increases in the enzyme protein and mRNA of fatty acid cyclooxygenase in the epinephrine-treated cells. A rapid cAMP production caused by epinephrine was followed by increases in the activity and mRNA of cyclooxygenase. Both dibutyryl cAMP and 8-bromo-cAMP also increased the level of the cyclooxygenase activity and mRNA. These results suggest that cAMP produced by beta-adrenergic stimulation was responsible for the increased cyclooxygenase mRNA level leading to induction of the cyclooxygenase enzyme. Furthermore, the addition of prostaglandin E2 (the final arachidonate metabolite in the MC3T3-E1 cells) brought about a rapid synthesis of intracellular cAMP followed by increases in the enzyme protein and mRNA of cyclooxygenase.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗

Percutaneous fiberoptic angioscopy of the cardiac valves.

The feasibility of percutaneous translumial angioscopy of the cardiac valves was examined in eight patients with and in 11 patients without valvular disease. In eight of these patients, a guiding balloon catheter (9F) was introduced into the aortic root, a guide wire (0.014 or 0.025 inch) was introduced through the catheter into the left ventricle to prevent dislocation of the catheter, and a fiberscope (1.6 or 4.6F) was advanced to the distal tip of the catheter. The balloon was then inflated with carbon dioxide and was manipulated against the aortic valve; a body temperature heparinized saline was infused through the catheter for observation. Similarly, the balloon catheter was advanced transseptally into the left atrium for observation of the mitral valve in four patients. Also, the balloon catheter was advanced through the right femoral vein into the right atrium for observation of the tricupid valve in three patients. In patients with a normal aortic valve, the aortic cusp surface was smooth and white and the edges were sharp. They opened briskly during systole and coapted each other completely during diastole. In rheumatic aortic regurgitation, the cuspus were thick and blunt and their coaptation insufficiency was observed during diastole. In a patient with rheumatic AS, globular and yellow cusps were observed. Mitral valve leaflets were smooth and white in a patients without mitral valvular disease, while the leaflets were yellow, thick and irregular, and blood regurgitation from the left ventricle into the left atrium could be observed in two patients with rheumatic MSR. The process of opening and closure of a tricuspid valve was also observed in three patients without tricuspid valvular disease.(ABSTRACT TRUNCATED AT 250 WORDS)

Aged↗

Crystallization and preliminary X-ray studies of a Bacillus subtilis and Thermus thermophilus HB8 chimeric 3-isopropylmalate dehydrogenase.

A chimeric gene was constructed by fusing the Bacillus subtilis and Thermus thermophilus genes coding for 3-isopropylmalate dehydrogenase, and expressed in Escherichia coli. The chimeric enzyme was crystallized in a size suitable for X-ray structure analysis. The crystal has a space group of P3(1)21 or P3(2)21, a = b = 77.1 A and c = 158.3 A, which is isomorphous with that of the native enzyme from T. thermophilus.

3-Isopropylmalate Dehydrogenase↗

Nucleotide sequence of the gene encoding NADH dehydrogenase from an alkalophile, Bacillus sp. strain YN-1.

The gene encoding NADH dehydrogenase from an alkalophile, Bacillus sp., was cloned and sequenced. The cloned DNA fragment contained an open reading frame of 1,557 nucleotides which encodes a polypeptide composed of 519 amino acid residues (Mr 55,830). The predicted amino acid sequence was consistent with the partial amino acid sequences including the N-terminal and C-terminal sequences determined in a previous study. Sequence comparison with other flavoenzymes revealed high homology between the present dehydrogenase and Escherichia coli thioredoxin reductase.

Amino Acid Sequence↗

Molecular cloning and nucleotide sequence of 3-isopropylmalate dehydrogenase gene (leuB) from an extreme thermophile, Thermus aquaticus YT-1.

A gene (leuB) coding for 3-isopropylmalate dehydrogenase [EC 1.1.1.85] from an extreme thermophile, Thermus aquaticus YT-1 was cloned in Escherichia coli and the nucleotide sequence was determined. It contains an open reading frame of 1,035 bp encoding 344 amino acid residues. The homology with that from T. thermophilus HB8 is 87.0% in nucleotide and 91.3% in amino acid sequences. No overlapped gene was found in the present leuB gene, in contrast to the previous prediction that Thermus leuD gene is overlapped with leuB [Croft et al. (1987) Mol. Gen. Genet. 210, 490-497]. Substitutions in the primary structure which are unique for the thermophile sequences are discussed in relation to the unusual stability of the thermophile dehydrogenase based on amino acid sequence comparison of 9 microorganisms including thermophiles and mesophiles.

3-Isopropylmalate Dehydrogenase↗

Crystallization and preliminary X-ray studies on Sulfolobus acidocaldarius ferredoxin.

Ferredoxin from a thermoacidophilic archaebacterium, Sulfolobus acidocaldarius, has been crystallized. The space group is P4(3)2(1)2 or P4(1)2(1)2 and the cell dimensions are a = b = 50.12 A and c = 69.52 A. The Vm value is calculated to be 1.88 A3/Da, assuming one molecule per asymmetric unit. The crystal diffracts X-rays beyond 2.0 A resolution.

Crystallization↗

Parathyroidectomy lowers blood pressure independently of changes in platelet free calcium.

Several lines of evidence indicate that the parathyroid gland plays a role in blood pressure regulation. To examine whether the attenuation of the development of hypertension by parathyroidectomy is associated with modifications in cellular calcium metabolism, intracellular free calcium was determined in fura-2-loaded platelets of spontaneously hypertensive rats after parathyroidectomy. Parathyroidectomy decreased systolic blood pressure (146 +/- 8 versus control 160 +/- 5 mmHg, n = 8 and 9, respectively; P less than 0.05) and serum ionized calcium (0.81 +/- 0.08 versus 1.28 +/- 0.04 mmol/l; P less than 0.01). However, no effect of parathyroidectomy on basal intracellular free calcium was detected (63.1 +/- 5.5 versus 65.0 +/- 5.1 nmol/l). Furthermore, there were no differences in the intracellular free calcium responses to thrombin in the presence and absence of external calcium, intracellular calcium discharge capacity or calcium extrusion from the cytosol. These results indicate that parathyroidectomy lowers blood pressure independently of alterations in platelet calcium handling.

Animals↗

The herald waves of influenza virus infections detected in Sendai and Yamagata cities in 1985-1990.

The community surveillance of respiratory virus infections performed during 1985-1987 in Sendai and 1988-1990 in Yamagata has identified a total of five herald waves of influenza virus infections: A/H3N2 virus infections in 1985 and 1989, A/H1N1 virus infections in 1986 and 1988, and type B virus infections in 1989. To investigate the antigenic and genetic relationships between the herald wave and epidemic strains, influenza A/H1N1 viruses isolated during the 1986 and 1988 herald waves were compared with those isolated during the 1986-1987 and 1988-1989 epidemic seasons, respectively, utilizing hemagglutination inhibition tests with anti-hemagglutinin monoclonal antibodies and oligonucleotide mapping of total viral RNAs. The results showed that multiple variants differing in antigenic and genetic properties were cocirculating during the 1986 herald wave as well as during each of the two epidemics (only one strain was isolated in the 1988 herald wave). It was also observed that viruses which had the antigenic and/or genetic characteristics closely similar to those of the viruses circulating in the 1986 and 1988 herald waves, were isolated during the winters of 1986-1987 and 1988-1989, respectively.

Antibodies, Monoclonal↗

Positive selection for uracil auxotrophs of the sulfur-dependent thermophilic archaebacterium Sulfolobus acidocaldarius by use of 5-fluoroorotic acid.

Uracil auxotrophs of Sulfolobus acidocaldarius were positively selected by using 5-fluoroorotic acid. The wild-type strain was unable to grow in medium containing 5-fluoroorotic acid, whereas the mutants grew normally. Positive selection could be done for the auxotrophs. Mutants deficient in orotidine-5'-monophosphate pyrophosphorylase activity were isolated.

Culture Media↗

Abnormal platelet and lymphocyte calcium handling in prehypertensive rats.

We have reported that the basal and stimulated cytosolic free calcium concentrations [( Ca2+]i) are elevated in platelets isolated from 12-14-week-old spontaneously hypertensive rats (SHR) as compared with normotensive Wistar-Kyoto (WKY) rats. To determine whether altered cell calcium metabolism precedes the development of overt hypertension, we measured [Ca2+]i under resting and stimulated conditions in blood platelets and thymic lymphocytes isolated from 4-week-old prehypertensive SHR and WKY rats. Blood pressure was similar in both groups (SHR 95 +/- 8 versus WKY rats 92 +/- 7 mm Hg). Basal [Ca2+]i in platelets was higher in SHR than WKY rats (63.4 +/- 3.9 versus 54.8 +/- 3.1 nM, p less than 0.003). Also the [Ca2+]i response to thrombin was greater in SHR than WKY rats in both the presence and absence of extracellular calcium. For lymphocytes, although no difference was detected in basal [Ca2+]i, the concanavalin A-induced peak [Ca2+]i was higher for SHR than WKY rats in both calcium-containing and calcium-free media. These results suggest that agonist-stimulated calcium influx and calcium discharge from intracellular stores are enhanced in both platelets and lymphocytes of 4-week-old SHR. We conclude that abnormalities in calcium metabolism in two different cell types precede the development of overt hypertension in the SHR.

Animals↗

Rise and fall of agonist-evoked platelet Ca2+ in hypertensive rats.

We previously reported an enhanced peak response of intracellular free Ca2+ to thrombin in platelets of spontaneously hypertensive rats in comparison with normotensive Wistar-Kyoto rats. In the present study, we compared the platelet intracellular Ca2+ response to the receptor-linked agonist thrombin with the response to the Ca2+ ionophore ionomycin. Basal intracellular Ca2+ was higher in hypertensive platelets as was leakage of fura-2. We confirmed the previous finding that the thrombin-induced intracellular Ca2+ peak is greater in platelets of hypertensive rats and noted that the rate of recovery from peak intracellular Ca2+ is significantly greater in this model. In contrast, the peak platelet intracellular Ca2+ response to ionomycin (50 nM and 5 microM) was not different between the two strains, and the rate of recovery from the peak response was only slightly depressed in hypertensive rats after the low dose of ionomycin. Internal Ca2+ discharge capacity, assessed by the intracellular Ca2+ response to a maximal dose of ionomycin in Ca(2+)-free medium, was not different between platelets of the two strains. Thus, activated platelet intracellular Ca2+ is not altered in the hypertensive rat when the nonphysiological ionophore ionomycin is used as agonist. However, a heightened intracellular Ca2+ response is observed when the receptor-mediated agonist thrombin is used. These results are consistent with the hypothesis that differences in receptor-linked second messenger pathways underlie the altered intracellular Ca2+ response in platelets of genetically hypertensive rats and may contribute to differences both in the mobilization of Ca2+ and in its fall.

Animals↗

Hemodynamic response to dietary sodium loading in essential hypertension studied with Doppler echocardiography.

The cardiac response to dietary salt loading was assessed by Doppler echocardiography during various sodium intakes (52-345 mEg per day) in 30 patients with essential hypertension. The Mitral flow velocity integral in the rapid filling phase (IntR) and the atrial contraction phase (IntA) was measured from the transmitral flow pattern, and the sum of IntR and IntA (IntR + IntA), the ratio of IntA to IntR (IntA/IntR), cardiac output (CO) and total peripheral resistance (TPR) were calculated. With salt loading, the mitral flow pattern remained almost unchanged in the nonsalt-sensitive (NSS) patients. Fourteen of the 19 salt-sensitive (SS) patients showed significant increases in IntR + IntA and CO with salt loading (IntR + IntA, from 13.9 +/- 2.8 to 17.9 +/- 3.6 cm, p less than 0.01; CO, from 6021 +/- 2130 to 8305 +/- 1699 ml/min, p less than 0.01), and were termed "salt-sensitive CO-dependent" (SS [COdep]), suggesting that the apparent pressor response to sodium loading was mediated by an increased CO. In the remaining five SS patients termed "salt-sensitive CO-independent" (SS [COindep]), IntA/IntR increased significantly with sodium repletion (from 0.66 +/- 0.23 to 0.90 +/- 0.31, p less than 0.01), without a significant change in IntR + IntA. Increments in IntA/IntR observed in the SS [COindep] patients were considered to be due to an elevation of total peripheral resistance (TPR), since changes in IntA/IntR were significantly correlated with those in TPR in all subjects (r = 0.617, p less than 0.01).

Adult↗

Recombinant human erythropoietin, but not iron supplementation, improves anemia in rats with adjuvant-induced arthritis.

Pathophysiological and therapeutic properties of anemia in rats with adjuvant-induced arthritis (AA) were investigated. Both anemia and chronic inflammation were induced in rats by a single injection of Freund's complete adjuvant. This study confirmed other earlier data that these anemic rats with AA had reduced serum iron levels and that the anemia was characterized as mild, non-progressive, hypochromic, microcytic. In addition, our studies showed that these anemic rats had slightly but significantly enhanced erythropoietin titers, but not renal failure; there was no significant difference in blood urea nitrogen and creatinine levels in anemic and normal groups. The anemia in rats with AA was improved by recombinant human erythropoietin (r-HuEPO) at 30 and 100 U/kg/day, given i.v. for 5 days. In contrast, iron-chondroitin-sulfate colloid (10 mg/kg/day, i.v. for 5 days) failed to improve the anemia and to enhance the effects of r-HuEPO. These data suggest that anemia in rats with adjuvant-induced arthritis is distinguished, pathophysiologically and therapeutically, from iron deficiency anemia, hemolytic anemia, and renal anemia.

Anemia, Hypochromic↗

Percutaneous fiberoptic cardioscopy of the left ventricle.

The feasibility of percutaneous transluminal cardioscopy of the left ventricle was examined in 35 patients with or without various heart diseases. A guiding balloon catheter (7 or 9 French) was introduced through the right femoral artery into the left ventricle. The balloon was inflated, and a fiberscope (1.6 or 4.3 French) was advanced to the distal tip of the catheter. The balloon was then manipulated against the portion of the ventricle targeted for examination, and heparinized saline (10 U/ml) at body temperature was infused through the catheter at 5 ml/sec for 3 to 6 sec. The images were recorded on videotape and 16 mm cinefilm. In 4 patients without demonstrable heart disease, the endocardial surface of the left ventricle was brown and the trabeculae became prominent during systole. The chordae connected to the mitral leaflets were white. In 3 of 5 patients with dilated cardiomyopathy, the endocardium was light yellow with thin trabeculae which barely contracted during systole. The endocardium was red or reddish-brown in 3 patients with acute myocarditis. In patients with rheumatic mitral stenosis white patches were scattered on the endocardial surface. In 7 of 8 patients the akinetic or dyskinetic segments representing prior infarctions were white. No complications other than transient ventricular arrhythmias were noted. We conclude that percutaneous fiberscopic imaging with a guiding balloon catheter is feasible and safe, and yields highly detailed images of the endocardium.

Acute Disease↗

Clinical application of modified polyurethane graft to blood access.

Based on a study of 34 clinical implantations, a porous poly(ether-urethane) graft developed for blood access was evaluated. Although it became clear that the graft has several advantages over expanded polytetrafluoroethylene (E-PTFE), it was also revealed that the occlusion at the anastomotic site was more frequently observed than that with E-PTFE. To improve the patency rate, a small modification of the luminal surface was attempted to give a better anchoring of the neointima. The results so far are better than those with the unmodified graft. However, the number of cases is small and the period of observation is short. More work will be necessary to confirm these preliminary results.

Arteriovenous Shunt, Surgical↗