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T Oshima

Publications and source records attributed to T Oshima.

At least 469 records · Page 26Linked to original sources

Identification of the domain of Saccharomyces cerevisiae adenylate cyclase associated with the regulatory function of RAS products.

Various truncated CYR1 genes of Saccharomyces cerevisiae were fused to efficient promoters and expressed in Escherichia coli and S. cerevisiae cells with or without the RAS genes. The catalytic domain of adenylate cyclase encoded by the 3'-terminal 1.3 kb region of the open reading frame of the CYR1 gene produced cyclic AMP, irrespective of the presence of RAS genes. The product of the 3'-terminal 2.1 kb region of CYR1 showed guanine nucleotide-dependent adenylate cyclase activity and produced a large amount of cAMP in the presence of the RAS gene. Thus, the domain encoded by the 0.8 kb region adjacent to the catalytic domain is associated with the regulatory function of the RAS products. The cyr1 RAS1 RAS2 cells carrying the 3'-terminal 1.3 kb region of CYR1 were unable to respond to environmental signals such as sulfur starvation and temperature shift, but the cyr1 cells carrying the 2.1 kb region and at least one RAS gene were able to respond to these signals. The environmental signals may be transferred to the adenylate cyclase system through the RAS products.

Adenylyl Cyclases↗

Anisakiasis - is the sushi bar guilty?

Since 1972, Japan and some other countries such as the USA have seen a sudden increase in cases of anisakiasis (infection with Anisakis sp.) and codworm an isakiasis (infection with the codworm, Pseudoterranova [= Phocanema] decipiens). Larval stages of these nematodes occur in the muscles of a variety of fish and squid, and ingestion of raw and undercooked fish or squid is the prime route of human infection. Thus the Japanese style of eating sashimi (sliced raw fish fillet) and the increasing popularity of sushi bars where such dishes are served, has often been suggested as contributing to the spread of anisakid infections. In this article, Tomoo Oshima discusses the current status of anisakiasis and codworm anisakiasis, showing that the apparent increase in human cases is related more to advances in diagnosis than to the commercialization of sushi.

Journal Article↗

A thermostable Gm-methylase recognizes the tertiary structure of tRNA.

The efficiency of methylation of tRNA by a thermostable tRNA(guanosine-2')-methyltransferase (Gm-methylase) was examined at various temperatures using several species of tRNA isolated from Escherichia coli, yeast and Bacillus subtilis, each possessing different thermal properties. The optimal temperature for the methylation reaction was ca. 20 degrees C lower than the melting temperature of the tRNA in each case. Arrhenius plots of the methylation reactions with various tRNAs gave straight lines below the optimal temperatures in all cases, with similar activation energies of between 10 and 14 kcal/mol. Above the optimal temperatures, the methyl acceptor activity decreased as the incubation temperature was raised to 80 degrees C, at which point the methylase was still active. A correlation was observed between the remaining methyl acceptor activity and the hyperchromicity of tRNA. These results suggest that Gm-methylase recognizes the tertiary structure of tRNA, and it is not the substrate tRNA but the enzyme which is activated by heat.

Base Sequence↗

Purification and properties of NADH dehydrogenase from a thermoacidophilic archaebacterium, Sulfolobus acidocaldarius.

An NADH dehydrogenase was purified to electrophoretical homogeneity from Sulfolobus acidocaldarius, a thermoacidophilic archaebacterium optimally growing at pH 2-3 and 75 degrees C. A 2,100-fold purification was achieved. The purified enzyme is an acidic protein with an isoelectric point of 5.6 and a molecular weight of 95,000, consisting of two 50,000-dalton subunits. The enzyme showed an absorption spectrum characteristic of flavoproteins, with maxima at 272, 372, and 448 nm. The enzyme is highly thermostable, is specific for NADH as an electron donor, and is capable of using 2,6-dichlorophenolindophenol, ferricyanide, benzoquinone, and naphthoquinone as electron acceptors. Though at a low rate, caldariellaquinone, a unique and sole benzothiophenequinone in the genus Sulfolobus, was also reduced by the enzyme, suggesting that the enzyme is a possible member of the respiratory chain of the thermoacidophilic archaebacterium.

Amino Acids↗

Purification and properties of the ATPase solubilized from membranes of an acidothermophilic archaebacterium, Sulfolobus acidocaldarius.

A novel ATPase was solubilized from membranes of an acidothermophilic archaebacterium, Sulfolobus acidocaldarius, with low ionic strength buffer containing EDTA. The enzyme was purified to homogeneity by hydrophobic chromatography and gel filtration. The molecular weight of the purified enzyme was estimated to be 360,000. Polyacrylamide gel electrophoresis of the purified enzyme in the presence of sodium dodecyl sulfate revealed that it consisted of three kinds of subunits, alpha, beta, and gamma, whose molecular weights were approximately 69,000, 54,000, and 28,000, respectively, and the most probable subunit stoichiometry was alpha 3 beta 3 gamma 1. The purified ATPase hydrolyzed ATP, GTP, ITP, and CTP but not UTP, ADP, AMP, or p-nitrophenylphosphate. The enzyme was highly heat stable and showed an optimal temperature of 85 degrees C. It showed an optimal pH of around 5, very little activity at neutral pH, and another small activity peak at pH 8.5. The ATPase activity was significantly stimulated by bisulfite and bicarbonate ions, the optimal pH remaining unchanged. The Lineweaver-Burk plot was linear, and the Km for ATP and the Vmax were estimated to be 1.6 mM and 13 mumol Pi.mg.-1.min-1, respectively, at pH 5.2 at 60 degrees C in the presence of bisulfite. The chemical modification reagent, 7-chloro-4-nitrobenzo-2-oxa-1,3-diazole, caused inactivation of the ATPase activity although the enzyme was not inhibited by N,N'-dicyclohexylcarbodiimide, N-ethyl-maleimide, azide or vanadate. These results suggest that the ATPase purified from membranes of S. acidocaldarius resembles other archaebacterial ATPases, although a counterpart of the gamma subunit has not been found in the latter. The relationship of the S. acidocaldarius ATPase to other ion-transporting ATPases, such as F0F1 type or E1E2 type ATPases, was discussed.

4-Chloro-7-nitrobenzofurazan↗

A microplate method for isolation of viruses from infants and children with acute respiratory infections.

Between December 1984 and December 1986, a microplate technique was adopted for isolation of viruses from infants and children with acute respiratory infections. By using two kinds of tissue culture microplates, i.e., the HHVM plate, containing human embryonic fibroblast (HEF), HEp-2, Vero and MDCK cells, and the MK plate which contains secondary monkey kidney cells, 1,080 field viruses were isolated from 1,061 (24.9%) out of 4,254 throat swabs. Of these 1,080 isolates, 1,003 (92.9%) were recovered in the HHVM plates and the remaining 77 (7.1%) in the MK plates. With the HHVM plate, influenza A and B viruses were cultivated in MDCK, RS virus in HEp-2, parainfluenza and mumps viruses in Vero, adenoviruses in both HEF and HEp-2, polioviruses in HEF, HEp-2 and Vero, coxsackie B viruses in both HEp-2 and Vero, rhino and echo viruses in HEF, herpes simplex virus in both HEF and HEp-2, and cytomegalovirus in HEF, although MK were more sensitive than Vero to parainfluenza and coxsackie B viruses. There was no difference in the rate of isolation of viruses between the microplate and ordinary tube methods. Cross contamination in the microplates was negligible for routine work.

Acute Disease↗

Influence of albumin on granulocyte and macrophage colony-forming efficiency.

Mouse granulocyte and macrophage precursors were assayed in plasma clot and fibrin clot cultures, and the effect of bovine serum albumin (BSA) on colony formation was investigated. The number of granulocyte colonies (CFU-g) and clusters increased as the albumin concentration was increased and the number of macrophage colonies (CFU-m) and clusters concomitantly decreased. The albumin-mediated suppression of macrophage colony formation was overcome by the addition of more than 10% fetal bovine serum (FBS) to the plasma clot culture. The effect of BSA and fatty-acid-free BSA on colony-forming efficiency was also tested in fibrin clot cultures containing 10% FBS. Both BSA and fatty-acid-free BSA at a final concentration of 0.5-2% enhanced CFU-g colony formation, while both forms of BSA reduced the number of CFU-m colonies. However, neither BSA nor fatty-acid-free BSA had any effect on colony formation in FBS-free fibrin clot cultures, and only BSA enhanced colony formation when transferrin, linoleic acid, alpha-thioglycerol and dextran were added to the culture. The number of CFU-g (15.6 +/- 3.1) was higher in cultures containing BSA, transferrin, etc., than the number (9.8 +/- 2.5) in cultures without BSA and including transferrin, etc., than the number (9.8 +/- 2.5) in cultures without BSA and including transferrin, etc. (p less than 0.01). The number of CFU-m (32.0 +/- 6.8) in cultures containing BSA and the other four factors was lower than the number (72.2 +/- 5.6) in the culture without BSA (p less than 0.001).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Intralymphocytic sodium and free calcium concentration in relation to salt sensitivity in patients with essential hypertension.

In order to clarify the relation between salt sensitivity and changes in intracellular sodium ([Na]i) and free calcium concentration ([Ca2+]i) after salt loading, [Na]i and [Ca2+]i were determined in lymphocytes of twenty patients with essential hypertension under a low salt diet (3 g/day) and a high salt diet (20 g/day) for seven days, respectively. They were classified as "salt-sensitive" (n = 10) or "nonsalt-sensitive" (n = 10) on the basis of the changes in blood pressure after salt loading. Both lymphocytic [Na]i and [Ca2+]i were significantly increased with salt loading in salt-sensitive patients (p less than 0.05 for both), while they were not affected by salt loading in nonsalt-sensitive patients. Lymphocytic [Ca2+]i showed a positive correlation with lymphocytic [Na]i under both low salt diet (r = 0.62, p less than 0.01) and high salt diet (r = 0.70, p less than 0.01) in all patients in both groups. In addition, a close and positive correlation was observed between the changes in lymphocytic [Na]i and those in lymphocytic [Ca2+]i after salt loading in all patients in both groups (r = 0.80, p less than 0.001). These results suggest that the increase in [Ca2+]i, possibly linked with the increase in [Na]i, may be involved in elevation of blood pressure in the salt-sensitive patients after salt loading.

Adult↗

[Determination of various tumor markers, with special reference to neuron-specific enolase in lung cancer].

Serum neuron-specific enolase (NSE) was measured in 23 patients with small cell lung cancer (SCLC) and 184 patients with non-small cell lung cancer (non-SCLC), both of which were untreated. Increased levels of serum NSE were observed in 82.6% (19/23) of SCLC, whereas 9.8% (18/184) of non-SCLC had positive results showing an overall positive rate of 17.9% (37/207) in lung cancer cases. In addition, the elevation of serum NSE levels in non-SCLC patients seemed to suggest poor prognosis. Elevated serum NSE levels returned to normal with either surgical resection of the tumor or response to chemotherapy, after which serum NSE levels were again raised to levels higher than the previous ones in cases of relapse or progression. The evaluation of serum NSE may be a useful marker for both diagnosis and monitoring of responsiveness to therapy as well as for recognition of relapse and progression in SCLC. Identification of NSE as assessed by immunohistochemical procedure employing the ABC method on formalin-fixed paraffin-embedded tissue sections in lung cancer cases of each histological type, showed that some materials from non-SCLC cases were positively stained despite the presence of normal serum NSE levels, and did not always parallel the serum levels. Among other various tumor markers determined, serum CA 19-9 had a relatively high positive rate of 38.2% (42/110) in adenocarcinoma of the lung.

Antigens, Neoplasm↗

[Foreign bodies of unknown origin found in a serosal hematoma between the stomach and the pancreas of a patient].

An autopsy of a 55-year-old man who had died suddenly revealed extensive intra-abdominal bleeding and a serosal hematoma between the stomach and the pancreas, in which numerous foreign bodies were found. They were located just adjacent to pancreatic tissue which had undergone changes of acute necrotizing pancreatitis. It was assumed that they had been shaped by a pathological mechanism contributing much to the sudden death of the patient. The nature of the foreign bodies was investigated by comparing their microscopic sections with those of various living organisms and by testing their chemical character with some histochemical methods. However, they had no similarities to known foreign bodies of animal or plant origin found in pathological sections of human tissues and no conclusive evidence on their chemical nature could be obtained either. Our unified opinion is that the foreign bodies might have been formed in human tissues in situ as a result of the pathological tissue changes. The morphological features of the bodies are described in detail and their significance is discussed.

Acute Disease↗

Stable integration of woodchuck hepatitis virus DNA in transplanted tumors and established tissue culture cells derived from a woodchuck primary hepatocellular carcinoma.

The fate of integrated woodchuck hepatitis viral (WHV) DNA was systematically investigated in DNA samples from primary hepatocellular carcinoma (HCC) of woodchucks, solid tumors transplanted in athymic mice derived from a primary HCC of woodchuck, and an established cell line of tissue culture originating from the transplanted tumor. In four of five woodchuck primary HCCs, WHV DNA integration was demonstrated in addition to various amounts of extrachromosomal replicative intermediate WHV DNA. The integration pattern of the primary HCCs does not indicate a common integration site on the host chromosome. The integration pattern in the established cells is identical to that in the transplanted tumor and similar but slightly different from that of the primary HCC. No extrachromosomal or replicative intermediates of WHV DNA were detected in the transplanted tumors or in the established cells of tissue culture. There are three integration sites on the chromosomes of the established cells. Results of Southern hybridization and restriction maps of cloned fragments suggest that all of these integrated WHV DNA sequences are not a complete genome but a part of the genome. A small portion corresponding to the cohesive region of the genome was not detected in all of these integrated WHV DNA. A positive role of WHV DNA integration on the generation of HCC is strongly suggested by the high incidence of WHV DNA integration in woodchuck primary HCCs and the stable maintenance of a certain mode of WHV DNA integration in the hepatoma-derived cell populations during passages of transplantation or serial growth of tissue culture.

Animals↗

Membrane-bound ATPase of a thermoacidophilic archaebacterium, Sulfolobus acidocaldarius.

The membranes of Sulfolobus, a thermoacidophilic archaebacterium showed two types of ATP hydrolyzing activity. One was that of a neutral ATPase at an optimum pH around 6.5. This enzyme was activated by 10 mM sulfate with a shift of optimum pH to 5. In these respects, the enzyme was similar to membrane-bound ATPase of Thermoplasma, another thermoacidophilic archaebacterium, reported by Searcy and Whatley [1982) Zbl. Bakt. Hyg., I. Abt. Orig. C3, 245-257). The enzyme hydrolyzed ATP and other NTPs, but not ADP or AMP. It was highly thermostable, but irreversibly inactivated in 0.1 M HCl. The other activity was that of an acidic apyrase at an optimum pH around 2.5. This enzyme was extremely stable toward high temperature and acid and inhibited by sulfate. Both of these ATP hydrolyzing enzymes were resistant to N,N'-dicyclohexylcarbodiimide (DCCD), azide, oligomycin, N'-ethylmaleimide, p-chloromercuribenzoate, orthovanadate, or ouabain. Sulfolobus ATPases differ from F1 and other transport ATPases so far described.

Adenosine Triphosphatases↗