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Biomedical subjects

J Ohta

Publications and source records attributed to J Ohta.

At least 109 records · Page 6Linked to original sources
PubMed1990

Visualization of sialoglycoproteins in polyacrylamide gels by acidic ninhydrin reaction.

A new method for staining sialoglycoproteins in polyacrylamide gel after disc electrophoresis is described. The method utilizes the reaction of sialic acids with an acidic ninhydrin reagent which yields a stable color with an absorption maximum at 470 nm. After electrophoresis, the polyacrylamide gel is placed in a test tube and heated with 5 ml of the acidic ninhydrin reagent for 10 min in a boiling water bath. Sialoglycoproteins are detected as brown bands. No additional procedure such as destaining is necessary. When 20 micrograms fetuin, a sialoglycoprotein, per gel is applied, the band remains visible for at least 2 h. Stained gel can be scanned with a gel scanner at 470 nm. When the stained gel was dried on a sheet of polypropylene filter, the color was stable for at least one month. The present method is superior to the method using Stains-all (3,3'-diethyl-9-methyl-4,5,4',5'-dibenzothiacarbocyanine) in specificity and simplicity for the detection of sialoglycoproteins.

Electrophoresis, Polyacrylamide Gel↗
PubMed1990

Formation of sulfate from L-cysteine in rat liver mitochondria.

Formation of sulfate in rat liver mitochondria was studied. About 0.1 mumol of sulfate was formed in mitochondria from 1 g of liver in 60 min when 10 mM L-cysteine was used as the substrate. Addition of either 10 mM 2-oxoglutarate or 10 mM glutathione to this system increased sulfate formation 3 to 4 times. The addition of both 2-oxoglutarate and glutathione resulted in a 20-fold increase in sulfate formation. Sulfate formation in the presence of 5 mM L-cysteine was 58% of that with 10 mM L-cysteine. L-Cysteine-glutathione mixed disulfide was not a good substrate, indicating that this mixed disulfide was not an intermediate of sulfate formation in the present system. Incubation of 3-mercaptopyruvate with rat liver mitochondria also resulted in sulfate formation, and the addition of glutathione accelerated it. Formation of sulfite and thiosulfate was also detected. These results indicate that sulfate is produced in mitochondria, at least in part, from L-cysteine through the transamination pathway (3-mercaptopyruvate pathway).

Animals↗
PubMed1990

[Combination chemotherapy with etoposide, ADM, and CDDP (EAP) for advanced gastric cancer].

EAP therapy has been performed on 50 cases of advanced gastric cancer from January 1988 to September 1989. Adriamycin 20 mg/m2, Cisplatin 40 mg/m2 and Etoposide 100 mg/m2 were administered on day 1 and 7, 2 and 8, and 4, 5 and 6, respectively, with not less than 2 courses every 3 to 4 weeks. Complete success, PR, NC and PD were obtained in 48, 21, 20 and 7 cases, respectively, the rate of effectiveness being 43.8% with a confidence interval 95% of 30-58%. The rate of effectiveness by lesions for evaluation was high (30.4, 100, and 50% for primary lesion, Virchow's lymphnodal metastasis and liver metastasis, respectively). MST was 5.1 months for EAP therapy, which was highly effective but led to no prolonged survival period. Thus, it is thought that good control of leukopenia, a dose limiting factor remains to be examined.

Adult↗
PubMed1990

Comparisons of interval breast cancers with other breast cancers detected through mass screening and in outpatient clinics in Japan.

In a nation-wide collaborative study on mass screening for breast cancer, we collected 152 cases of interval breast cancer diagnosed at 35 hospitals or clinics distributed throughout Japan. The definition of interval breast cancer used in the present study is "breast cancer cases which were diagnosed as having 'no malignant findings' in a previous screening for breast cancer but subsequently diagnosed as 'breast cancer' at a hospital or medical clinic within two years of the previous screening." The clinical stages and prognoses of these interval cancer were analyzed and compared with those of other breast cancers detected through mass screening and in outpatient clinics. In the clinical staging of interval breast cancer, Tis (non infiltrating cancer) accounted for only 2.1%, compared to 8.0% in cases detected through mass screening. At stage I 43.4% were interval breast cancers compared to 32.9% breast cancers detected through mass screening and 25.4% diagnosed in outpatient clinics. The stage differences between interval breast cancers and breast cancers detected through mass screening were not statistically significant. Five-year survival rates were 85.6% for interval breast cancers, 91.7% for breast cancers detected through mass screening and 84.7% for breast cancers diagnosed in outpatient clinics. Ten-year survival rates were 75.9, 80.5 and 78.1%, respectively, suggesting the interval breast cancer cases to show a similar prognosis to that of breast cancer cases diagnosed in outpatient clinics. The differences in five- and 10-year survival rates among the three groups were not statistically significant. From the present study we were not able to confirm the general belief of interval cancer being more aggressive in nature and showing a poorer prognosis than cancer detected through periodic screening. The reasons for this are discussed.

Adult↗
PubMed1989

Expression of UDP-glucose: poriferasterol glucosyltransferase in the process of differentiation of a true slime mold, Physarum polycephalum.

An expression of UDP-glucose:poriferasterol glucosyltransferase activity associated with differentiation of a true slime mold, Physarum polycephalum, from haploid myxoamoebae to diploid plasmodia was demonstrated. In the haploid cells, this enzyme activity was not detected, but after conjugation of the myxoamoebae, the enzyme activity was expressed and increased definitely. In the plasmodial stage, high enzyme activity was maintained constantly. The enzyme was partially purified (35-fold purification, and 28% yield), and molecular weight of 72,000, pH optimum of 7.0, and some characteristics were demonstrated.

Glucosyltransferases↗
PubMed1989

DNA synthesis by the isolated nuclear matrix from synchronized plasmodia of Physarum polycephalum.

Nuclear matrices were isolated from plasmodia of a true slime mold, Physarum polycephalum, and the DNA synthetic activity in vitro was examined. These matrices isolated in S-phase catalyzed DNA synthesis requiring Mg2+, deoxyribonucleoside 5'-triphosphates and ATP, without exogenous templates. The activity changed during S-phase with the rate of in vivo DNA replication. Product analysis by gel electrophoresis revealed that the matrices produced Okazaki fragments. These results suggest that DNA synthesis partially reflects in vivo DNA replication. DNA synthesis was sensitive to aphidicolin, heparin and N-ethylmaleimide, indicating involvement of the alpha-like DNA polymerase of Physarum. Exogenous addition of activated DNA stimulated DNA synthesis 4-10-fold and suggested that only some of the existing enzymes are involved in endogenous DNA synthesis. Matrices isolated in G2-phase were also associated with a similar DNA synthetic activity, but they did not produce Okazaki fragments in vitro. It is, therefore, concluded that nuclear matrices are associated with alpha-like DNA polymerase throughout the cell cycle, and that some of the enzymes participate in in vivo DNA replication in S-phase; thus, DNA replication is possibly controlled by this process. The relationship between DNA synthetic activities by the isolated nuclei and matrices was also discussed.

Adenosine Triphosphate↗
PubMed1989

Reduction of 3-mercaptopyruvate in rat liver is catalyzed by lactate dehydrogenase.

It has been assumed that the in vivo reduction of 3-mercaptopyruvate, an intermediate of cysteine metabolism, to 3-mercaptolactate is catalyzed by lactate dehydrogenase (EC 1.1.1.27) though no definitive evidence has been presented. In order to examine this assumption, reduction of 3-mercaptopyruvate and its inhibition were studied using rat liver homogenate, lactate dehydrogenase purified from rat liver and anti-lactate dehydrogenase antiserum. Reduction of 3-mercaptopyruvate was actively catalyzed by rat liver homogenate and by the purified lactate dehydrogenase. This reducing activity was completely inhibited by anti-lactate dehydrogenase antiserum. These results indicate that the reduction of 3-mercaptopyruvate to 3-mercaptolactate in rat liver is catalyzed by lactate dehydrogenase.

Animals↗
PubMed1989

[New selection criterion for Fontan procedure: pulmonary artery clamping test and pulmonary vascular resistance in increased pulmonary blood flow].

A new selection criterion for Fontan procedure, pulmonary artery clamping test (PACT) was developed and employed in 13 candidates for Fontan procedure. PACT was aimed to evaluate the response of pulmonary vascular bed to increased pulmonary blood flow and calculate the pulmonary vascular resistance in the increased pulmonary blood flow as a selection criterion for Fontan procedure, preoperatively. After a median sternotomy incision, an electromagnetic flow meter was employed for measuring the pulmonary blood flow (Qp) on the pulmonary trunk. By clamping the left pulmonary artery and the right pulmonary artery, right pulmonary vascular resistance and the left pulmonary resistance in the pulmonary blood flow of Qp were calculated respectively. By means of electrical analogue, right and left lungs were simulated as resistors in parallel. And the equivalent pulmonary vascular resistance Rpc was calculated as the predicted pulmonary vascular resistance in the increased pulmonary blood flow of two times Qp. Fontan procedure was performed in 9 cases with a Rpc of less than 3 unit.m2 with a successful result and no death. Postoperative cardiac index ranged from 2.6 to 3.7 L/min.m2 with a mean of 2.8 L/min.m2. Postoperative pulmonary vascular resistance ranged from 1.6 to 3.2 unit.m2. Postoperative cardiac index was correlated with postoperative pulmonary vascular resistance. And postoperative pulmonary vascular resistance was well correlated with equivalent pulmonary vascular resistance but was not correlated with preoperative pulmonary vascular resistance measured by Fick's method at the preoperative catheterization. In Fontan procedure, the pulmonary blood flow frequently increases to a certain degree postoperatively. And pulmonary vascular resistance is not a static resistance, but a dynamic resistance. Hence, in discussing a pulmonary vascular resistance as a selection criterion for Fontan procedure, the pulmonary blood flow in which the pulmonary vascular resistance stand should be taken into account. Equivalent pulmonary vascular resistance Qpc is a theoretical selection criterion for Fontan procedure and well reflects the postoperative pulmonary vascular resistance. Fontan procedure can be successfully performed with a Rpc of less than 3 unit.m2.

Adolescent↗
PubMed1987

Expression of poriferasterol monoglucoside associated with differentiation of Physarum polycephalum.

A glycolipid which was expressed during a differentiation from haploid myxoamoebae to diploid plasmodia of a true slime mold, Physarum polycephalum, has been examined. In the amoeboid stage, cells did not contain this glycolipid, but after conjugation of the haploid cells, this substance appeared and increased in its amount. From structural studies of the purified glycolipid, it has been identified as poriferasterol monoglucoside.

Cell Differentiation↗
PubMed1987

DNA synthesis in isolated nuclei from synchronized plasmodia of Physarum polycephalum.

Nuclei were isolated from synchronized plasmodia of a true slime mold, Physarum polycephalum, in S-phase, and DNA synthesis in the nuclei was studied in vitro. The nuclei catalyzed DNA synthesis at the rate of 0.7 ng DNA/1.0 X 10(6) nuclei/30 min at 25 degrees C, which was 5 times higher than that catalyzed in G2-phase nuclei. The DNA synthesis required Mg2+, four kinds of deoxyribonucleoside 5'-triphosphates and ATP, suggesting that the mode of synthesis is a replicative-type, but not a repair-one. Sedimentation analysis of the DNA products revealed that the nuclei produced 2-4S DNA fragments mainly during a 30-sec pulse incubation, and 2-4S, 5-12S and longer fragments during a 15-min incubation. The pulse- and chase-labeling experiments showed that the 2-4S fragments shifted discontinuously to longer fragments. These results indicate that the nuclei catalyze the formation of 2-4S Okazaki fragments first and then their subsequent ligation. Eighty % and 96% of the DNA synthesis was inhibited by 200 micrograms/ml aphidicolin and 40 mM N-ethylmaleimide, respectively, but 80% of the activity was resistant to 100 microM 2',3'-dideoxythymidine 5'-triphosphate. These results suggest that the DNA synthesis is catalyzed by the alpha-type DNA polymerase of Physarum polycephalum.

Aphidicolin↗
PubMed1987

Mass screening for colorectal cancer by testing for occult blood under restricted diet and a questionnaire in Osaka.

Because of the recent increase in the incidence of colorectal cancer in Japan, attempts were made to find an effective method for screening asymptomatic patients with this cancer. A total of 12,520 participants in Osaka were screened in a 2-year program from April 1982 by a test of fecal occult blood under a restricted diet, and a medical questionnaire. Occult blood in feces was examined for 3 consecutive days using one guaiac-impregnated slide (Shionogi B) each day. This slide was about twice as sensitive as the Hemoccult slide. Further diagnostic workups were required in 3,452 individuals, of which 2,602 (75.7%) were due to positive occult blood. Proctosigmoidoscopy was performed in 2,214 (64.4%) of these persons, barium enema in 1,397 persons, and flexible colonoscopy in 187 persons. Colorectal cancers were detected in 18 patients (0.14% of the total screened); ten of these cancers were in an early stage. Thus this screening method is reliable for detecting asymptomatic cancer of the colon and rectum.

Colonic Neoplasms↗
PubMed1987

Effect of L-2-oxothiazolidine-4-carboxylate administration on glutathione and cysteine concentrations in guinea pig liver and kidney.

Effect of L-2-oxothiazolidine-4-carboxylate administration on the glutathione and cysteine concentrations in liver and kidney was studied in the guinea pig. Liver glutathione concentration increased significantly by 21 to 29% at one to three hours after the intraperitoneal injection of 5 mmol of sodium L-2-oxothiazolidine-4-carboxylate per kg of body weight. Cysteine concentration did not change significantly. In contrast to the liver, a significant increase in cysteine and also a significant decrease in glutathione concentrations were observed in the kidney. Incubation of L-2-oxothiazolidine-4-carboxylate with liver and kidney homogenates resulted in cysteine formation of 1.21 and 0.56 mumol per g of fresh tissue per 30 min, respectively. These results seem to indicate that, in the liver, L-2-oxothiazolidine-4-carboxylate administration resulted in the formation of cysteine, which was utilized for glutathione synthesis. In the kidney, it seems to be suggested that the administration of this compound accelerated glutathione turnover.

Animals↗
PubMed1986

Mass screening for colorectal cancer by testing fecal occult blood.

The validity of screening for colorectal cancer by testing for occult blood on 2 successive days was evaluated over a 2-year period beginning April 1980 by testing 9449 individuals without symptoms. The Shionogi slide (Shionogi Pharmaceutical Co., Osaka, Japan), a commercial guaiac-impregnated slide with moderate sensitivity, was used for screening subjects under dietary restriction. Of the 1401 persons (14.8%) who had a positive reaction for occult blood, 858 (61.2%) received further diagnostic examinations, and 265 of them proved to have one or more abnormalities of the gastrointestinal tract. Colorectal cancer was detected in 11 persons and polyps in 91 persons. Eight of the cancers were in an early stage. This screening method was found to be suitable for large-scale mass screening, and appears to have high diagnostic value.

Adult↗
PubMed1985

Visualization method for sulfolipids and its application to the determination of nanomole quantities of lipid sulfur.

A simple and sensitive visualization method for sulfolipids on a thin-layer chromatogram is described. By spraying with an acidic solution of azure A, a complex was formed between an anionic sulfolipid and a blue cationic compound. After the unbound dye was washed out by brief soaking in methanol, sulfolipids were visualized as clear dark-blue bands on a light-blue background. As little as 0.5 nmol could be detected. Sulfolipid-dye complex was estimated by densitometry or colorimetric measurement after extraction with chloroform/methanol. For the quantitative determination of sulfolipids having long sugar chains, it is necessary to treat thin-layer chromatography plates with acetic anhydride before color development. Of the other tissue lipids not containing sulfuric acid ester that were tested none were stained significantly. A linearity of quantitative determination was observed over the range of 1-8 nmol.

Acetylation↗