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H Yamano

Publications and source records attributed to H Yamano.

At least 19 recordsLinked to original sources
PubMed1999

Carrier-mediated transport systems for glucose in mucosal cells of the human oral cavity.

The in vitro uptake study was performed using the isolated cells of human oral mucosa, buccal and the dorsum of the tongue, to investigate the mechanisms of glucose uptake. The uptake of D-glucose was much larger in cells of the dorsum of the tongue than in buccal cells and was inhibited more extensively by 2-deoxy-D-glucose, a substrate of facilitative glucose transporters, than by alpha-methyl-D-glucoside, a specific substrate of SGLT1, suggesting the larger contribution of a facilitative transporter than Na(+)/glucose cotransporter. Furthermore, from the results of inhibition studies by several sugar analogues including maltose and D-mannose, GLUT1 and/or GLUT3 were suggested to take part in the glucose uptake by oral mucosa. Therefore, we have attempted to confirm the expression of glucose transporters on the oral mucosa by employing Western blotting. As a result, it was suggested that SGLT1, GLUT1, GLUT2, and GLUT3 are expressed in the epithelial cells of human oral mucosa.

Absorption↗
PubMed1998

The role of the destruction box and its neighbouring lysine residues in cyclin B for anaphase ubiquitin-dependent proteolysis in fission yeast: defining the D-box receptor.

Programmed proteolysis of proteins such as mitotic cyclins and Cut2/Pds1p requires a 9-residue conserved motif known as the destruction box (D-box). Strong expression of protein fragments containing destruction boxes, such as the first 70 residues of Cdc13 (N70), inhibits the growth of Schizosaccharomyces pombe at metaphase. This inhibition can be overcome either by removal of all lysine residues from N70 using site-directed mutagenesis (K0-N70) or by raising the concentration of intracellular ubiquitin. Consistent with the idea that competition for ubiquitin accounts for some of its inhibitory effects, wild-type N70 not only stabilized D-box proteins, but also Rum1 and Cdc18, which are degraded by a different pathway. The K0-N70 construct was neither polyubiquitinated nor degraded in vitro, but it blocked the growth of strains of yeast in which anaphase-promoting complex/cyclosome (APC/C) function was compromised by mutation, and specifically inhibited proteolysis of APC/C substrates in vivo. Both K0-N70 and 20-residue D-box peptides blocked polyubiquitination of other D-box-containing substrates in a cell-free ubiquitination assay system. These data suggest the existence of a D-box receptor protein that recognizes D-boxes prior to ubiquitination.

Amino Acid Sequence↗
PubMed1998

Hepatitis C virus 1b(II) infection and development of chronic hepatitis, liver cirrhosis and hepatocellular carcinoma: a case-control study in Japan.

This study was performed to determine the distribution of hepatitis C virus (HCV) genotypes among asymptomatic carriers (ASC) and patients with chronic hepatitis without cirrhosis (NC-CH), liver cirrhosis (LC), and hepatocellular carcinoma (HCC) in the Osaka area, and to assess whether infection with HCV genotype 1b (II) is more likely to develop into hepatocellular carcinoma (HCC) than is that with genotype 2a (III) or genotype 2b (IV). Genotypes of all study subjects were determined by Okamoto's method. HCV genotype 1b was detected in 100 of the 143 ASC (69.9%), 551 of the 726 NC-CH patients (75.9%), 86 of the 103 patients with LC (83.5%), and 153 of the 179 HCC (85.5%) patients. Using unconditional logistic regression analysis, the age- and sex- adjusted odds ratios contrasting NC-CH with ASC, LC with ASC and HCC with ASC were 1.38 [95% confidence interval (CI) = 0.93-2.05], 2.28 (95% CI = 1.12-4.63) and 2.27 (95% CI = 1.02-5.06) respectively. HCV genotype 1b is predominant in both healthy carriers and patients with chronic liver diseases in the Osaka area. The findings from the three case-control studies indicate that type 1b infection is more closely associated with the development of LC and HCC than type 2a or 2b through its role in the progression of chronic liver inflammation to a cirrhotic stage.

Adult↗
PubMed1998

The risk of hepatitis C virus infection among blood donors in Osaka, Japan.

To estimate the risk of hepatitis C virus (HCV) infection among blood donors, we conducted a retrospective cohort study with 448,020 HCV-seronegative donors who donated blood more than once between February 1992 and July 1997 in Osaka (a total of 2,676,738 allogeneic blood donations). The donors were divided into four age groups according to the age at the initial donation: Group A (16-24 years), Group B (25-34 years), Group C (35-49 years) and Group D (50-64 years). Fifty-nine donors became infected with HCV among the 448,020 HCV-seronegative donors who donated blood more than once within a period of approximately five years. In a total of 1,095,668 person-years of observation (PYO), the incidence rate was 5.38 per 105 PYO, with the 95% confidence interval (95% C.I.) being 4.10 to 6.95. There was no significant difference in the incidence rate between males and females. Young donors between the ages of 16 and 24 (8.89; 95% C.I., 6.04 to 12.61) had a significantly higher incidence rate of HCV infection than donors between the ages of 35 and 49 (1.81; 0.67 to 3.95). The cumulative risk of HCV infection among donors between the ages of 16 and 64 was estimated to be 0.27% (95% C.I., 0.16 to 0.39) for males and 0.27% (95% C.I., 0.15 to 0.38) for females. Based on the recent age-specific incidence rate, the cumulative risk of HCV infection among blood donors was estimated to be about 0.3% in the Osaka district of Japan. The incidence rate differed among age groups, indicating that HCV infection is associated with age-related behaviors and the need for further epidemiological research towards the eradication of community-acquired HCV infection.

Adolescent↗
PubMed1997

Fission yeast Cut2 required for anaphase has two destruction boxes.

The fission yeast Schizosaccharomyces pombe cut2(+) gene is essential for sister chromatid separation. Cut2 protein, which locates in the interphase nucleus and along the metaphase spindle, disappears in anaphase with the same timing as mitotic cyclin destruction. This proteolysis depends on the APC (Anaphase-Promoting Complex)-cyclosome which contains ubiquitin ligase activity. The N-terminus of Cut2 contains two stretches similar to the mitotic cyclin destruction box. We show that both sequences (33RAPLGSTKQ and 52RTVLGGKST) serve as destruction boxes and are required for in vitro polyubiquitination and proteolysis. Cut2 with doubly mutated destruction boxes inhibits anaphase, whereas Cut2 with singly mutated boxes can suppress cut2 mutations. Strong expression of the N-terminal 73 residues containing the destruction boxes leads to the accumulation of endogenous cyclin and Cut2, and arrests cells in metaphase, whereas the same fragment with the mutated boxes does not. Cut2 proteolysis occurs in vitro using Xenopus mitotic extracts in the presence of functional destruction boxes. Furthermore, Cut2 is polyubiquitinated in an in vitro system using HeLa extracts, and this polyubiquitination requires the destruction boxes.

Anaphase↗
PubMed1996

The role of proteolysis in cell cycle progression in Schizosaccharomyces pombe.

A cell-free system derived from Xenopus eggs was used to identify the 'destruction box' of the Schizosaccharomyces pombe B-type cyclin, Cdc13, as residues 59-67: RHALDDVSN. Expression of indestructible Cdc13 from a regulated promoter in S.pombe blocked cells in anaphase and inhibited septation, showing that destruction of Cdc13 is necessary for exit from mitosis, but not for sister chromatid separation. In contrast, strong expression of a polypeptide comprising the N-terminal 70 residues of Cdc13, which acts as a competitive inhibitor of destruction box-mediated proteolysis, inhibited both sister chromatid separation and the destruction of Cdc13, whereas an equivalent construct with a mutated destruction box did not. Appropriately timed expression of this N-terminal fragment of Cdc13 overcame the G1 arrest seen in cdc10 mutant strains, suggesting that proteins required for the initiation of S phase are subject to destruction by the same proteolytic machinery as cyclin.

Amino Acid Sequence↗
PubMed1996

Cut2 proteolysis required for sister-chromatid seperation in fission yeast.

Although mitotic cyclins are well-known substrates for ubiquitin-mediated proteolysis at the metaphase-anaphase transition, their degradation is not essential for separation of sister chromatids; several lines of evidence suggest that proteolysis of other protein(s) is required, however. Here we report the anaphase-specific proteolysis of the Schizosaccharomyces pombe Cut2 protein, which is essential for sister-chromatid separation. Cut2 is located in the nucleus, where it is concentrated along the short metaphase spindle. The rapid degradation of Cut2 at anaphase requires its amino-terminal region and the activity of Cut9 (ref. 14), a component of the 20S cyclosome/anaphase-promoting complex (APC), which is necessary for cyclin destruction. Expression of non-degradable Cut2 blocks sister-chromatid separation but not cell-cycle progression. This defect can be overcome by grafting the N terminus of cyclin B onto the truncated Cut2, demonstrating that the regulated proteolysis of Cut2 is essential for sister-chromatid separation.

Amino Acid Sequence↗
PubMed1996

Diagnosis of colorectal tumorous lesions by magnifying endoscopy.

BACKGROUND: The magnifying colonoscope allows 100-fold magnified viewing of the colonic surface. METHODS: We examined 2050 colorectal tumorous lesions by magnifying endoscopy, stereomicroscopy, and histopathology and classified these lesions according to pit pattern. Based on stereomicroscopy, lesions with a type 1 or 2 pit pattern were nontumors, whereas lesions with types 3s, 3L, 4, and/or 5 pit patterns were neoplastic tumors. RESULTS: The pit patterns observed by magnifying endoscopy were fundamentally similar to those demonstrated in stereomicroscopic images. When the diagnosis by magnifying endoscopy was compared with the stereomicroscopic diagnosis, there was agreement in 1130 of 1387 lesions (81.5%). True neoplasms could be differentiated from non-neoplastic lesions. Of lesions with a type 5 pit pattern with a bounded surface, 11 of 22 (50%) were found to be invasive cancers with involvement of the submucosal layer. If this pit pattern is found to involve a relatively broad area of the mucosal surface, extensive malignant invasion (sm-massive) should be strongly suspected. CONCLUSIONS: The magnifying colonoscope provides an accurate instantaneous assessment of the histology of colorectal tumorous lesions. This may help in decision making during colonoscopy.

Adenoma↗
PubMed1994

Phosphorylation of dis2 protein phosphatase at the C-terminal cdc2 consensus and its potential role in cell cycle regulation.

We show that the fission yeast dis2 protein phosphatase, which is highly similar to mammalian type 1 phosphatase, is a phosphoprotein containing phosphoserine (phospho-S) and threonine (phospho-T). It has several phosphorylation sites, two of which locate in the C-terminus. Phospho-T was abolished in the alanine substitution mutant at the C-terminal T316, which is conserved as a residue in the cdc2 consensus, TPPR, in a number of type 1-like phosphatases. In G2-arrested cdc2-L7 cells, the degree of T316 phosphorylation was reduced, whereas it was enhanced in metaphase-arrested nuc2-663 mutant cells. Phospho-T was produced in dis2 by fission yeast cdc2 kinase, but not in the substitution mutant A316, indicating that the T316 residue was the site for cdc2 kinase in vitro. Phosphatase activity of wild type dis2 was reduced by incubation with cdc2 kinase, but that of mutant dis2-A316 was not. Phosphorylation of T316 hence has a potential significance in cell cycle control in conjunction with cdc2 kinase activation and inactivation. Overexpression phenotypes of wild type dis2+, sds21+ and mutant dis2-A316, sds21-TPPR genes were consistent with negative regulation of dis2 by phosphorylation. This type of regulation would explain why cells harboring the dis2-11 mutation enter mitosis but fail to exit from it.

Amino Acid Sequence↗
PubMed1994

Prevalence of second generation antibody to hepatitis C virus among voluntary blood donors in Osaka, Japan.

To clarify the demographic characteristics of the prevalence of hepatitis C virus (HCV) infection in Osaka, Japan, where hepatocellular carcinoma is common, we investigated the screening data of antibody to HCV (anti-HCV, DAINABOTHCVPHA, second generation assay) in 197,600 voluntary blood donors residing in Osaka. The study found that age-standardized prevalence of anti-HCV was significantly higher than that of HBsAg (2.25 cf 0.86 percent among males, P < 0.001; 2.17 cf 0.55 percent among females, P < 0.001. It was much higher in the blood donors aged 55-64 years than in those aged 16-54 years (8.49 cf 1.32 percent among males, P < 10(-5); 7.26 cf 1.42 percent among females, P < 10(-5)). The prevalence of anti-HCV among males was significantly higher than that of females in the younger (25-34 years) generations (1.02 to 1.49 percent cf 0.71 to 1.13 percent, P < 0.05). A similar tendency was observed in the prevalence of high-titer (> or = 2(12)) anti-HCV. The number of coinfection (both HBsAg and anti-HCV seropositive) was very small, and it was not statistically different from the expected number.

Adolescent↗
PubMed1994

Cumulative risk of hepatocellular carcinoma in hepatitis C virus carriers: statistical estimations from cross-sectional data.

The probability of developing hepatocellular carcinoma (HCC) among hepatitis C virus (HCV) carriers during their life-time is unknown. This paper addresses the estimation of the cumulative risk of HCC among HCV carriers using cross-sectional data. Age-specific prevalences of HCV carriers among the general population were estimated according to 5-year age group, based on the data of 2nd-generation anti-HCV assay in blood donors resident in Osaka (33,226 males and 29,054 females). Seropositivity of anti-HCV among 422 HCC cases, and the Osaka Cancer Registry data on HCC were used in the estimations of 5-year age-specific incidence rates of HCV-linked HCC. Using these data, the cumulative risk, i.e., the probability of contracting HCC within the following 15 years in 50-year-old HCV carriers, was estimated as 28% for males and 6% for females.

Carcinoma, Hepatocellular↗
PubMed1993

Negative regulation of mitosis by the fission yeast protein phosphatase ppa2.

To understand the role of the type 2A-like protein phosphatase in the cell division cycle, we investigated the mutant phenotypes obtained when the fission yeast ppa1+ and ppa2+ phosphatase genes (which encode polypeptides with approximately 80% identity to mammalian type 2A phosphatases) were either deleted or overexpressed. We also investigated the in vivo effect of okadaic acid, an inhibitor of protein serine/threonine phosphatases, on cell division. We show that ppa2+ interacts genetically with the cell cell regulators cdc25+ and wee1+, as a ppa2 deletion is lethal when combined with wee1-50 but partially suppresses the conditional lethality of cdc25-22 mutation. Evidence that ppa2+ negatively controls the entry into mitosis, possibly through the regulation of cdc2 tyrosine phosphorylation, is presented. ppa2 phosphatase is abundant in the cytoplasm, in contrast to the type 1-like phosphatase dis2, which is enriched in the nucleus. Overproduced ppa1 or ppa2 proteins accumulate in the cytoplasm near the nuclear periphery, and cells arrest in interphase. Okadaic acid-treated cells, like a ppa2 deletion, are short in length and display protein hyperphosphorylation. Cytokinesis is also inhibited, producing binucleated cells. We show that ppa2 is the genetic locus controlling okadaic acid sensitivity. The ppa2 deletion reveals the same hyperphosphorylated proteins as okadaic acid. When a strain deleted for ppa2 is treated with okadaic acid, cell size is reduced further to that of wee1-50 mutant strain or overexpressing the cdc25+ gene product, suggesting functional relationship of ppa2 with the cdc25 tyrosine phosphatase and/or the wee1 kinase in cell cycle control.

Base Sequence↗
PubMed1992

Protein phosphatases and cell division cycle control.

Fission yeast has at least ten protein phosphatase genes that appear to play distinct roles in cell cycle control. Because of functional overlap, a clear lethal phenotype can be obtained only after multiple genetic alterations. Cells that have lost the protein phosphatase 1 (PP1)-like dis2/sds21 phosphatase activities prematurely enter mitosis and remain in a defective mitotic state with high H1 kinase activity and without sister chromatid disjunction. The same phenotype can be obtained in the presence of hydroxyurea. Overexpression of PP1-like phosphatase, on the other hand, delays the entry into mitosis. Cells that have lost PP2A-like ppa2 phosphatase activity also prematurely enter mitosis with a reduction in cell size. This semi-wee phenotype is enhanced in delta ppa2 mutants treated with the phosphatase inhibitor, okadaic acid. Genetic interactions between ppa2 and mitotic regulators suggest that ppa1/ppa2 phosphatase may directly or indirectly inhibit p34cdc2/cyclin kinase. Thus both PP1- and PP2A-like phosphatases in fission yeast may negatively regulate entry into mitosis. The major property of the dis2/sds21 mutant which is distinct from those of the ppa2/ppa1 mutant is its failure to inactivate the p34cdc2/cyclin complex after entry into mitosis. A novel phosphatase regulator encoded by sds22+ binds to dis2 phosphatase and controls the substrate specificity which appears to become essential in the progression from metaphase to anaphase.

Cell Division↗
PubMed1992

Gene cloning of Porphyromonas gingivalis specific antigens recognized by serum of adult periodontitis patient.

1. Porphyromonas gingivalis is believed an important pathogen of adult periodontitis. A gene library of P. gingivalis 381 was constructed in lambda phage vector L47.1. The library was probed with serum obtained from patients of severe adult periodontitis. Two clones, lambda MDBG101 and lambda MDBG103 which were expressed, 200 and 160 kDa respectively, were selected and further studied. 2. The expressed antigens in these two clones were also reacted with rabbit antiserum against whole cells, capsular fraction and cell surface fraction of P. gingivalis. 3. Genes coding protein antigens in lambda MDBG101 and lambda MDBG103 were subcloned into high-copy-number plasmid vector pACYC184 and subclones obtained were designated as MD101 and MD103. Recombinant plasmids, pMD101 and pMD103, differed in their restriction endonuclease digestion. 4. Immunodiffusion analysis showed that cloned proteins from MD101 and MD103 reacted with antiserum against P. gingivalis but did not react with antiserum against Prevotella intermedia, Prevotella loescheii and Prevotella asaccharolyticus. 5. These data suggest that P. gingivalis species-specific antigens has been successfully cloned and expressed in Escherichia coli. Since these cloned specific antigens were recognized by adult periodontitis patient sera, the recombinant antigen will be useful material for the development of serodiagnosis of P. gingivalis infection in adults periodontitis.

Adult↗
PubMed1992

Study on the interaction between soybean beta-amylase and substrate by the stopped-flow method.

The hydrolysis of substrates (maltoheptaose, maltopentaose, and maltotetraose) catalyzed by soybean beta-amylase [EC 3.2.1.2] at pH 5.4 and 25 degrees C was followed by monitoring small changes in the quenching of fluorescence due to tryptophan residues by the stopped-flow method. By analysis of whole time course, the dissociation constants, KdS, of enzyme-substrate and enzyme-product complexes were reasonably evaluated; and the difference in fluorescence intensities per mol between the enzyme-complex (ES or EP) and the free enzyme, delta F, was determined. The molecular activity, k0, was also determined by a new method of half time analysis. The KdS and k0 values are in good agreement with our kinetic data reported previously. The delta Fs of substrates were of smaller magnitude than those of products (G2 and G3), which means that the higher the binding affinity of the ligand is, the smaller the delta F value is. This indicates that at least two tryptophan residues must be located in the active site if the enzyme is rigid, or that if there is only one, the active site must undergo a structural change caused by the binding of ligand.

Hydrolysis↗