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Biomedical subjects

H Shibata

Publications and source records attributed to H Shibata.

At least 361 records · Page 20Linked to original sources
PubMed1994

Kinetic studies on the plasminogen activation by the staphylokinase-plasmin complex.

A pure complex of staphylokinase and plasmin was prepared by affinity chromatography with lysine-Sepharose, which enabled the simple analysis of the mechanism of plasminogen activation by staphylokinase. We used a truncated staphylokinase (SAK), which lacks the 10 amino acid residues at the NH2 terminal of native staphylokinase. The purity of this complex was confirmed by the native PAGE profile. Image analysis of the SDS-PAGE profile revealed that the molar ratio of plasmin and SAK in the complex was about 1:1. Using this SAK-plasmin complex, the kinetic parameters for the activation of Glu- or Lys-plasminogen were determined. The kinetic constant, kcat/Km, obtained when Lys-plasminogen was used as a substrate was approximately 10 times higher than that obtained when Glu-plasminogen was used. This plasminogen activation property of the SAK-plasmin complex was comparable to that of other plasminogen activators, such as streptokinase, urokinase, and tissue-type plasminogen activator (t-PA). This SAK-plasmin complex will simplify the elucidation of plasminogen activation by SAK. Through kinetic studies, the fibrin specificity and participation of plasminogen activator inhibitor will be clarified.

Fibrinolysin↗
PubMed1994

Amino acid sequence of an inhibitor from the silkworm (Bombyx mori) hemolymph against fungal protease.

A novel protein protease inhibitor (FPI-F) which is highly specific for fungal proteases and subtilisin was isolated from the silkworm hemolymph, and its amino acid sequence was determined by conventional methods. The inhibitor consisted of 55 amino acid residues and had a molecular weight of 6,100. The inhibitor included eight cysteine residues and relatively large amounts of acidic amino acids, but neither alanine, methionine nor tryptophan. The amino acid sequence of FPI-F was not homologous with those of other known protease inhibitors of microbe, plant or animal origin.

Amino Acid Sequence↗
PubMed1994

Purification and characterization of alanine dehydrogenase from a cyanobacterium, Phormidium lapideum.

Alanine dehydrogenase (AlaDH) was purified to homogeneity from cell-free extracts of a non-N2-fixing filamentous cyanobacterium, Phormidium lapideum. The molecular mass of the native enzyme was 240 kDa, and SDS-PAGE revealed a minimum molecular mass of 41 kDa, suggesting a six-subunit structure. The NH2 terminal amino acid residues of the purified AlaDH revealed marked similarity with that of other AlaDHs. The enzyme was highly specific for L-alanine and NAD+, but showed relatively low amino-acceptor specificity. The pH optimum was 8.4 for reductive amination of pyruvate and 9.2 for oxidative deamination of L-alanine. The Km values were 5.0 mM for L-alanine and 0.04 mM for NAD+, 0.33 mM for pyruvate, 60.6 mM for NH4+ (pH 8.7), and 0.02 mM for NADH. Various L-amino acids including alanine, serine, threonine, and aromatic amino acids, inhibited the aminating reaction. The enzyme was inactivated upon incubation with pyridoxal 5'-phosphate (PLP) followed by reduction with sodium borohydride. The copresence of NADH and pyruvate largely protected the enzyme against the inactivation by PLP.

Alanine Dehydrogenase↗
PubMed1994

The presence of low molecular weight germinative substances in Bacillus cereus T spore extract.

An extract from intact spores of Bacillus cereus T having a germination-inducing activity was studied. Two distinct germinative principles were found through dialysis of the extract. One was diffusible through the dialysis membrane and the other was non-diffusible. The activity of the former fraction was inhibited by the addition of 1 mM glycoletherdiamine-N,N,N',N'-tetraacetic acid (GEDTA), whereas the latter fraction was inactive unless GEDTA was added to the assay system. The diffusible principle maintained the major portion of the activity found in the crude spore extract. By means of high-performance liquid chromatography (HPLC) using a gel permeation chromatography column, 9 fractions were obtained from the deproteinized diffusible fraction. Of those fractions, two fractions (No. 1 and No. 8) were responsible for the germination-inducing activity, but no reconstituted activity was observed unless both fractions No. 1 and No. 8 were added to the assay system. Amino acid analysis of fraction No. 1 revealed that the fraction was rich in free amino acids, especially in alanine. On the other hand, by the use of reverse-phase HPLC and fast atom bombardment mass spectrometry, it was concluded that the effective substance in fraction No. 8 was inosine. Based on these findings, it was suggested that the active substances in fraction No. 1 might be a free amino acid such as L-alanine and/or Ca2+ and a Ca(2+)-binding substance.

Alanine↗
PubMed1994

Manganese ions induce tonic contraction after relaxation in a high-K+ medium in ileal longitudinal smooth muscle of guinea-pig.

In ileal longitudinal muscle 5 mM Mn2+ inhibited completely the K+ (60 mM)-induced tonic tension to the base line; however, the tension progressively increased to above the level of original tonic response evoked by K+ after 3 h in the presence of Mn2+. Tetrodotoxin 5 x 10(-5) M) had no influence on the tension development in the presence of Mn2+ in the high-K+ medium. Mn2+ also increased the tension in a high-K+, Ca(2+)-free medium. The Ca2+ antagonist, gallopamil (10(-6) M) inhibited the development of tension in the presence of Mn2+ in the high-K+ medium. The 45Ca uptake determined by the lanthanum method remained unchanged from control levels after 3 h of the 5 mM Mn2+ application in the high-K+ medium in spite of the development of the tension. The manganese uptake in the high-K+ medium, increased in accordance with the increase of duration of 5 mM Mn2+ application. Gallopamil inhibited manganese uptake in the high-K+ medium. These results suggest that Mn2+ firstly reduces K(+)-induced tension by inhibition of Ca2+ influx, subsequently, Mn2+ ions accumulate in the intracellular compartments through voltage-operated Ca2+ channels and may activate contractile proteins in the ileal muscle.

Animals↗
PubMed1994

Immunoglobulin M and G antibodies in mice in response to Toxoplasma gondii (S-273) infection and their antigen recognition patterns in western blotting on various post-infection days.

Immunoglobulin M and G antibody responses in mice experimentally infected with Toxoplasma gondii (S-273) and the reaction patterns with T. gondii (RH) tachyzoite antigens were studied on various post-infection days (PIDs) (2nd to 36th PIDs) using a commercially available IgM and IgG enzyme linked immunosorbent assay (ELISA) test systems and Western blotting (WB) technique. IgM antibody in ELISA test appeared to be positive on 12th PID (absorbance 0.764) and reached its peak level on 16th PID (absorbance 1.338) showing a slow decline thereafter with an absorbance of 0.800 even on 36th PID. Positivity of IgM was confirmed by WB except for 36th PID. IgG appeared on 16th PID with an absorbance of 0.248 and showed a steady increasing tendency even on 36th PID (absorbance 1.747). However, IgG positivity on WB was observed only on 29th PID and afterwards. On Western blots, both IgM and IgG showed interesting antigen recognition patterns on various PIDs. At the most IgM recognised seven antigens of 14kDa to 53kDa while IgG recognised eight antigens of 17kDa to 53kDa. Major antigens recognised by IgM were of 53kDa and 21kDa while the major band recognised by IgG was of 19kDa. The major bands, however, showed variability in their consistency during various PIDs. All the antigens recognised by IgM and IgG were not identical.

Animals↗
PubMed1994

Seroprevalence of hepatitis B and C viruses in eastern Nepal.

The seroprevalence of hepatitis B virus (HBV) and hepatitis C virus (HCV) in 303 serum samples collected from that apparently healthy population inhabitating different areas in eastern Nepal was studied. Samples were collected at Dharan Municipality, Sunsari85), Pancha Kanya Village Development Committee, Ilam86), Dhankuta Hile, Dhankuta82) and Basantapur Village Development Committee, Tehrathm50). HBsAg and anti-HBsAg antibody was screened by reverse passive haemagglutination (RPHA) and passive haemagglutination (PHA) respectively and positivity was confirmed by enzyme linked immunosorbent assay (ELISA). Anti-HCV antibody was detected by ELISA. None of the samples were positive for HBsAg. Anti-HBsAg antibody was positive in 1.9% (6/303). The positive rate increased with age reaching 25% positivity among the elderly. The anti-HBsAg antibody positivity was 2.35, 2.32, 1.22 and 2.00 in Dharan, Ilam, Dhankuta and Tehrathum respectively Anti-HCV antibody was detected only in one sample (15-year-old boy) collected in Dharan. These findings indicate that the HBV and HCV infections are not active in eastern Nepal.

Adolescent↗
PubMed1994

Status of soil-transmitted helminthic infection in Nepal.

A retrospective evaluation of the status of soil-transmitted helminthic infection in Nepal during the period from 1985 to 1992 was conducted at the Parasitology Laboratory, Department of Pathology, TU Teaching Hospital in Kathmandu. An average of 6,537 faecal samples were examined each year for the presence of various types of intestinal parasites. The annual rate of positivity of these samples for soil-transmitted helminthiasis ranged from 18.0 to 36.6%. The annual incidence decreased every successive calendar year in both adults and children, irrespective of sex. Among the various types of helminths, the most common was Ascaris lumbricoides (roundworm), followed by hookworm. The incidence of A. lumbricoides remained constant throughout the study period, while that of other parasites markedly decreased each year.

Adult↗
PubMed1994

[Acidified aspirin-induced gastric lesion in rats with hepatic cirrhosis produced by N-nitrosodiethylamine or carbon tetrachloride. Effect of aldioxa on gastric lesions].

Gastric lesion was induced by the oral administration of acidified aspirin in rats with hepatic cirrhosis produced by N-nitrosodiethylamine (NDA) or carbon tetrachloride (CCl4). Gastric lesion by acidified aspirin was aggravated in NDA-induced cirrhosis, but not in CCl4-cirrhotic rats. To clarify this difference in the susceptibility of the gastric mucosa, gastric mucosal blood flow and gastric emptying were measured by the hydrogen gas clearance method and beads method, respectively. Gastric mucosal blood flow was lower and gastric emptying was significantly delayed in NDA-induced cirrhotic rats as compared with the controls, but not in CCl4-induced cirrhotic rats. Gastric mucosal blood flow in NDA-induced cirrhotic rats was significantly decreased by the oral administration of acidified aspirin as compared with the controls. Aldioxa dose-dependently inhibited the gastric lesion formation by acidified aspirin and inhibited the decrease of gastric mucosal blood flow in NDA-induced cirrhotic rats. These results suggest that aggravation of gastric lesion induced by acidified aspirin in NDA-induced cirrhotic rats would be due to the decrease of gastric mucosal blood flow and delay of gastric emptying. In addition, aldioxa showed a protective effect against gastric lesions induced by acidified aspirin in NDA-induced cirrhotic rats, suggesting that this compound would have an inhibitory effect on gastric lesions that are accompanied by hepatic cirrhosis.

Allantoin↗
PubMed1994

A comparative study of the enzymological features of alpha-amylase in the Drosophila melanogaster species subgroup.

A comparative study was done on the enzymological features of alpha-amylase among six sibling species belonging to the Drosophila melanogaster species subgroup. The pH optima were 7.6 for D. melanogaster, D. mauritiana and D. simulans and 7.4 for D. erecta, D. teissieri and D. yakuba. The temperature optima were commonly 37 C in all species studied. However, there are two types of temperature dependence; a strict one as observed in D. melanogaster, D. mauritiana and D. simulans, and a flat one as observed in D. erecta, D. teissieri and D. yakuba. These separations are consistent with the general phylogeny within this species subgroup. The substrate dependence was almost the same in all the species studied. The Vmax and Km were also estimated for each species. We found clear differences in the enzymological features between the species. Thus these differences might reflect an adaptation of the amylase enzyme system to speciation in this species subgroup.

Amylopectin↗
PubMed1994

Steroid contents and cortical steroidogenic enzymes in non-hyperfunctioning adrenal adenoma.

The recent increasing use of ultrasound and computed tomography has revealed numbers of incidentally discovered adrenal tumors. Many studies have focused on their surgical management, but the biological characteristics of these adrenal tumors have remained unclear. Adrenal tumors were resected from 10 patients who underwent gastrectomy or cholecystectomy. No signs or symptoms of adrenal hormone excess or deficiency were evident either before or after the operation. Moreover, after surgery, no major differences in signs and symptoms including blood pressure levels were observed. Before surgery, neurogenic tumors and cysts were excluded by enhanced magnetic resonance imaging. Steroid contents and both the activities and amounts of steroidogenic cytochrome P-450s in the adrenocortical adenomas of these patients were examined. Microscopic examination revealed that the tumors were surrounded by a thin, non-intact capsule; the surrounding cortex was not atrophic and apparently normal; and the cells of both the tumor and adjacent portions were arranged in nests and cords. Measurements of all steroid content (pregnenolone, progesterone, corticosterone, 11-deoxycorticosterone, 18-hydroxydeoxycorticosterone, cortisol, and dehydroepiandrosterone) except aldosterone in 5 resected adrenal tumors were within the normal ranges for the adrenals of 5 patients with renal cell carcinoma. Aldosterone content in tumor portions was significantly lower than in the apparently normal adrenals. Although in both tumor and adjacent portions of another 5 resected adrenal tumors the activities and amounts of cytochrome P-450s (P-450scc, P-450(11 beta), P-450aldo, P-450(17 alpha), and P-450c21) were also within the normal ranges, the activities of P-450scc and P-450(17 alpha) in the tumor portion were greater than those in the adjacent portion.(ABSTRACT TRUNCATED AT 250 WORDS)

Adrenal Cortex Neoplasms↗
PubMed1994

[Usefulness of hemoglobin A1c and fructosamine for screening of glucose intolerance in community adults].

The usefulness of hemoglobin A1c and fructosamine for screening of glucose intolerance were compared in 383 semi-urban residents ranging from 35 to 88 years of age. Both the sensitivity and specificity of the indices for screening of those with 110 mg/dl and over of fasting blood glucose were examined. The results showed that hemoglobin A1c was better than fructosamine for screening for glucose intolerance.

Adult↗
PubMed1994

[Changes in C100-3, and N14 antibodies in patients with chronic hepatitis C treated with interferon].

In interferon therapy for chronic hepatitis type C, we assayed C100-3 and N14 antibodies, and also determined IgM-C100-3 and IgM-N14 antibodies as well as HCV-RNA. N14 antibody was assayed by an end point titer method. In patients responding to interferon therapy, C100-3 and N14 antibody titers were decreased and became negative. In patients irresponsive to interferon therapy, C100-3 antibody titers were hardly changed, while N14 antibody titers were markedly increased. IgM-C100-3 and IgM-N14 antibody levels were significantly higher in the patients non-responding to interferon therapy than in those responding to interferon therapy. The daily assays of C100-3 and N14 antibody titers are considered useful in judging the effectiveness of interferon against chronic hepatitis type C.

Antigens, Viral↗
PubMed1994

Marked increase of CD8+S6F1+ and CD8+CD57+ cells in patients with graft-versus-host disease after allogeneic bone marrow transplantation.

Peripheral blood lymphocytes (PBL) from 24 allogeneic bone marrow transplant (BMT) recipients were studied serially using flow cytometry and two-color analysis. Dual labelling with two monoclonal antibodies (moAbs), CD8/S6F1 (CD11a) and CD8/CD57 was used to analyze the surface phenotypes of PBL after allogeneic BMT. In patients with acute and chronic GVHD, CD8+S6F1+ cells were markedly increased from the onset of GVHD and recovered to normal range 6 years after transplantation. By contrast, CD8+S6F1- cells fell below normal range and remained markedly decreased for 2-3 years after transplantation in patients with acute and chronic GVHD. A slight but significant increase of CD8+CD57- cells was observed with clinical signs of acute GVHD. On the other hand, CD8+CD57+ cells were increased after the onset of acute and chronic GVHD and recovered to normal range 6 years after transplantation. These results suggest that these subsets of CD8+ cells may play important roles in the pathophysiology of GVHD.

Adolescent↗
PubMed1994

Prevalence of HCV-antibodies and HCV-RNA in donor blood in Tokushima Prefecture.

The incidence of C100-3 among the blood specimens qualified for transfusion according to the conventional criteria was 1.1%. The incidence of C100-3 in donor blood in Tokushima Prefecture is not significantly different from that reported for all Japan. Of the donors positive for the conventional screening test and C100-3, 73.6% showed high ALT levels. For all antibodies, the incidence of HCV-RNA was very low in the donors positive for a single antibody, but was high in those positive for multiple antibodies. All of the donors showing the 3 antibodies were positive for HCV-RNA. While a test for multiple antibodies is thought to be effective for the screening of HCV, more blood needs to be discarded, having a serious cost-performance problem. The O.D. value for C100-3 and the 2nd antibody seem to be useful reference value for antibody titers.

Adolescent↗
PubMed1994

Seroepidemiological study of toxoplasmosis in two different geographical areas in Nepal.

A total of 302 serum samples collected from Chitawan (159) and Mustang (143) districts of Nepal were included in this study. Anti-toxoplasma antibody was detected using micro-latex agglutination (MLA) and ELISA methods. An overall positive rate was found to be 57.9%. The positive rate in Chitawan was significantly higher (64.1%) (less than 1,000 m altitude) compared to that in Mustang (51.0%) (more than 3,000 m altitude) (p < 0.05). Females in Chitawan showed significantly higher positive rate (71.2%) compared to males (56.9%) (p < 0.05). On the contrary, though insignificantly, males showed higher positive rate (57.9%) compared to that of females (43.3%) in Mustang. Almost equal positive rate was observed among males in both study area. Females in Chitawan showed significantly higher (71.2%) positive rate compared to their counterparts in Mustang (43.3%) (p < 0.001). A slight increase in positive rate with age was observed in Chitawan while in Mustang a decreasing trend was noticed. Ethnically though statistically not significant, Indo-Aryans showed a higher positive rate (69.2%) compared to the positive rate shown by Tibeto-Burmans (63.1%) in Chitawan while the reverse was true in Mustang (Tibeto-Burmans: 53.8% and Indo-Aryans: 38.4%). Interestingly, 2.9% and 1.3% of MLA positive samples showed toxoplasma IgM antibody. None of the IgM positive samples were positive for toxoplasmic antigens.

Altitude↗
PubMed1994

[The monitoring of cytomegalovirus antigenemia and arterial oxygen saturation for the early detection of cytomegalovirus pneumonia].

Eighteen patients underwent allogeneic bone marrow transplantation (allo. BMT) during the period May, 1991 to December, 1992 in the Center for Adult Diseases, Osaka. They were monitored for cytomegalovirus (CMV) antigenemia and arterial oxygen saturation (SaO2). More than 10 antigen-positive cells per 50,000 polymorphonuclear leukocytes were detected in five of 18 patients. Three of these 5 patients developed CMV pneumonia several weeks after the first detection of more than 10 positive cells. Six of 18 patients developed interstitial pneumonia (IP) (3 CMV pneumonia and 3 idiopathic IP). SaO2 decreased less than 95% several days before the development of IP in 3 of these 6 patients (2 of CMV pneumonia and 1 of idiopathic IP). CMV antigenemia assay and SaO2 assay were thus both considered to be useful for the early detection or prediction of development of CMV pneumonia.

Adolescent↗