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A Ohashi

Publications and source records attributed to A Ohashi.

At least 73 records · Page 4Linked to original sources
PubMed2000

Syntrophothermus lipocalidus gen. nov., sp. nov., a novel thermophilic, syntrophic, fatty-acid-oxidizing anaerobe which utilizes isobutyrate.

A new anaerobic, thermophilic, syntrophic, fatty-acid-oxidizing bacterium designated strain TGB-C1T was isolated from granular sludge in a thermophilic upflow anaerobic sludge blanket (UASB) reactor. The cells were slightly curved rods and were weakly motile. Spore formation was not observed. The optimal temperature for growth was around 55 degrees C and growth occurred in the range 45 to 60 degrees C. The pH range for growth was 5.8-7.5, and the optimum pH was 6.5-7.0. Crotonate was the only substrate that allowed the strain to grow in pure culture. However, in co-culture with the thermophilic, hydrogenotrophic Methanobacterium thermoautotrophicum strain delta H, the isolate could syntrophically oxidize saturated fatty acids with 4-10 carbon atoms, including isobutyrate. During the degradation of isobutyrate by the co-culture, isobutyrate was isomerized to butyrate, which was then oxidized. The strain was not able to utilize sulfate, sulfite, thiosulfate, nitrate, fumarate or Fe(III) as electron acceptor. The DNA base composition was 51.0 mol%. 16S rDNA sequence analysis revealed that the strain belongs to the family Syntrophomonadaceae, but it was only distantly related to other known species of beta-oxidizing syntrophs. Hence, the name Syntrophothermus lipocalidus is proposed for TGB-C1T as a new species of a new genus.

Bacterial Typing Techniques↗
PubMed2000

Cultivation and in situ detection of a thermophilic bacterium capable of oxidizing propionate in syntrophic association with hydrogenotrophic methanogens in a thermophilic methanogenic granular sludge.

The thermophilic, anaerobic, propionate-oxidizing bacterial populations present in the methanogenic granular sludge in a thermophilic (55 degrees C) upflow anaerobic sludge blanket reactor were studied by cultivation and in situ hybridization analysis. For isolation of propionate-degrading microbes, primary enrichment was made with propionate as the sole energy source at 55 degrees C. After several attempts to purify the microbes, a thermophilic, syntrophic, propionate-oxidizing bacterium, designated strain SI, was isolated in both pure culture and coculture with Methanobacterium thermoautotrophicum. Under thermophilic (55 degrees C) conditions, strain SI oxidized propionate, ethanol, and lactate in coculture with M. thermoautotrophicum. In pure culture, the isolate was found to ferment pyruvate. 16S ribosomal DNA sequence analysis revealed that the strain was relatively close to members of the genus Desulfotomaculum, but it was only distantly related to any known species. To elucidate the abundance and spatial distribution of organisms of the strain SI type within the sludge granules, a 16S rRNA-targeted oligonucleotide probe specific for strain SI was developed and applied to thin sections of the granules. Fluorescence in situ hybridization combined with confocal laser scanning microscopy revealed that a number of rod-shaped cells were present in the middle and inner layers of the thermophilic granule sections and that they formed close associations with hydrogenotrophic methanogens. They accounted for approximately 1.1% of the total cells in the sludge. These results demonstrated that strain SI was one of the significant populations in the granular sludge and that it was responsible for propionate oxidation in the methanogenic granular sludge in the reactor.

Bacteria↗
PubMed1999

Assessment of residual bile duct stones with use of intraductal US during endoscopic balloon sphincteroplasty: comparison with balloon cholangiography.

BACKGROUND: We sought to determine the utility of intraductal ultrasonography (IDUS) in detecting residual bile duct stones during endoscopic balloon sphincteroplasty. METHODS: Eighty-one consecutive patients with bile duct stones who underwent IDUS during endoscopic balloon sphincteroplasty were studied. IDUS was performed with a thin-caliber ultrasonic probe (diameter 2.0 mm, frequency 20 MHz) by a transpapillary route after stone extraction. When IDUS or balloon-retrograde cholangiography suggested residual stones, the bile duct was cleared again with a Dormia basket. Extraction of the stones was confirmed by direct duodenoscopic visualization. Videotapes of IDUS and cholangiograms were reviewed retrospectively without knowledge of the results of other diagnostic modalities. RESULTS: In 27 of 81 patients (33%), IDUS detected small residual stones not seen on cholangiography. When stones were fragmented with mechanical lithotripsy, the accuracy of IDUS in detecting small residual stones was significantly greater than that of balloon-endoscopic retrograde cholangiography (95% vs 50%, p < 0. 001). When the bile duct was greater than 10 mm in diameter, the accuracy of IDUS in detecting small residual stones was significantly greater than that of cholangiography (92% vs. 56%, p < 0.001). CONCLUSIONS: IDUS is useful for detecting small residual bile duct stones during endoscopic balloon sphincteroplasty when stones are fragmented by mechanical lithotripsy or when there is evidence of a dilated bile duct (>10 mm).

Adult↗
PubMed1999

Preoperative assessment of extrahepatic bile duct carcinoma using three-dimensional intraductal US.

BACKGROUND: We investigated the utility of a new imaging modality, three-dimensional intraductal ultrasonography (US), for staging bile duct cancer. METHODS: In eight patients with extrahepatic bile duct carcinoma, two- and three-dimensional intraductal US was used to assess tumor invasion of the right hepatic artery, portal vein, and pancreatic parenchyma before resection. The findings were correlated with histologic information from the resected specimen. RESULTS: Three-dimensional intraductal US enabled accurate assessment of tumor invasion of the right hepatic artery in 88% of cases, the portal vein in 100%, and pancreatic parenchyma in 100%. Two-dimensional intraductal US enabled accurate assessment of invasion of these structures in 88%, 88%, and 88% of cases. CONCLUSIONS: Three-dimensional intraductal US is useful in assessing tumor stage in bile duct carcinoma.

Aged↗
PubMed1999

How many biopsies should be performed during percutaneous transhepatic cholangioscopy to diagnose biliary tract cancer?

BACKGROUND: The sensitivity of biopsy in the diagnosis of cholangiocarcinoma using percutaneous transhepatic cholangioscopy is not well defined. METHODS: Patients with a biliary tract malignancy (n = 52) underwent directed biopsy during percutaneous transhepatic cholangioscopy using a 1.8 mm diameter forceps. Histologic findings were correlated with endoscopic appearance. RESULTS: A diagnosis of carcinoma was made in all four patients with a tumor of the major duodenal papilla and in all 15 patients with a polypoid bile duct tumor with two biopsies from the mass. In 19 patients with stenotic bile duct cancer, a positive diagnosis was made in 95% of cases when three biopsies were taken from the margin of the stenotic area. When cholangioscopy showed a tortuous, dilated vessel (n = 10), the diagnosis of cancer was made with two biopsies taken from the margin of the stenosis. In 14 patients with metastatic bile duct cancer, the diagnosis was made in only 43% of cases when three biopsies were taken from the margin of the stenosis. When combined with results from the three biopsies taken from within the area of stenosis, the sensitivity for diagnosing pancreatic cancer improved from 20% to 60%. CONCLUSIONS: Directed cholangioscopic biopsies are highly sensitive for the diagnosis of cholangiocarcinoma but less sensitive for cancer metastatic to the bile duct. The numbers and locations of the biopsies required to make a diagnosis of carcinoma depend on the origin and cholangioscopic appearance of the tumor.

Adult↗
PubMed1999

Expression of proopiomelanocortin, corticotropin-releasing hormone (CRH), and CRH receptor in melanoma cells, nevus cells, and normal human melanocytes.

Proopiomelanocortin (POMC) is a 31 kDa prohormone that is processed to various bioactive peptides, including adrenocorticotropin (ACTH), melanotropins (alpha, beta, gamma-MSH), lipotropins, and endorphins. POMC is expressed not only in the pituitary gland but also in a variety of nonpituitary organs and tumors, including melanomas. We previously showed that normal human melanocytes produce and secrete alpha-MSH and ACTH, and furthermore, that advanced melanoma cells generally produce higher amounts of POMC peptides that correlate with tumor progression. To elucidate the mechanism of this upregulation, the expression of genes encoding corticotropin-releasing hormone (CRH) and its receptor, CRH-R, as well as POMC and the MSH receptor (MC1-R), was evaluated by reverse transcriptase-polymerase chain reaction using cultured human melanoma cells, nevus cells, and normal melanocytes. Our results show that all melanocytic cells express CRH, CRH-R, POMC, and MC1-R, with highest intensities in melanoma cells. Furthermore, immunohistochemistry shows that CRH as well as POMC is strongly expressed in advanced melanomas, such as vertically growing lesions of acral lentiginous, nodular and metastatic melanomas, in contrast to negative expression in nevus cells. These results indicate that tumor progression accentuates CRH, CRH-R, and POMC expression by melanoma cells.

Corticotropin-Releasing Hormone↗
PubMed1999

The depigmenting effect of alpha-tocopheryl ferulate on human melanoma cells.

Oral vitamin E (alpha-tocopherol, alpha-T) supplementation has been reported to improve facial hyperpigmentation. alpha-Tocopheryl ferulate (alpha-TF) is a compound of alpha-T and ferulic acid connected by an ester bond; ferulic acid is also an antioxidant, and could scavenge free radicals induced by ultraviolet (UV) radiation, and thus maintain the long-lasting antioxidative effect of alpha-T. Our aim was to see whether alpha-TF might be useful as a whitening agent and an antioxidant to improve and prevent facial hyperpigmentation following UV exposure. In this study, the inhibitory effect of alpha-TF on melanogenesis was examined biochemically using human melanoma cells in culture. The results show that alpha-TF, solubilized in ethanol or in 0.5% lecithin, inhibited melanization significantly, as did alpha-T at a concentration of 100 microg/mL, without inhibiting cell growth. This phenotypic change was associated with inhibition of tyrosinase and 5, 6-dihydroxyindole-2-carboxylic acid polymerase activities, and the degree of inhibition was dose dependent. No significant effect on DOPAchrome tautomerase activity was observed. alpha-TF did not directly inhibit tyrosinase activity of the large granule fraction extracted from human melanoma cells, and Western blotting revealed that there were no changes in protein content or in molecular size of tyrosinase, tyrosinase-related protein (TRP)-1 or TRP-2. Therefore, the inhibition of tyrosinase activity by alpha-TF might be due to effects at the post-translational level, and possibly by a secondary molecule activated by alpha-TF. These results suggest that alpha-TF is a candidate for an efficient whitening agent which suppresses melanogenesis and inhibits biological reactions induced by reactive oxygen species.

Antioxidants↗
PubMed1999

Limitation of cholangiography in assessing longitudinal spread of extrahepatic bile duct carcinoma to the hepatic side.

BACKGROUND: Preoperative assessment of longitudinal spread of bile duct carcinoma (BDC) to the hepatic side remains a difficult problem for diagnostic imaging. METHODS: We studied the accuracy of cholangiography in assessing BDC. In 54 patients with extrahepatic bile duct cancer, cholangiographic findings were compared retrospectively with the histological findings of the resected specimens. RESULTS: Histological examination of specimens indicated longitudinal spread of the tumour to the hepatic side in 22 of 54 patients. The accuracy of cholangiography in assessing the extent of the longitudinal spread was only 34/54 (63%). When the cholangiographic images showed a main tumour with a collapsed edge, there was a significantly higher frequency of longitudinal spread compared with tumours with sharp edges (P< 0.05). In contrast, the accuracy of mapping biopsy under percutaneous transhepatic cholangioscopy (n=24) was 83%. CONCLUSIONS: Cholangiography cannot accurately assess the extent of the longitudinal spread of bile duct cancer. When cholangiographic images show a tumour with a collapsed edge, preoperative or intraoperative histological examination is essential to determine a suitable surgical line.

Adenocarcinoma↗
PubMed1999

Influence of bile duct diameter on the therapeutic quality of endoscopic balloon sphincteroplasty.

BACKGROUND AND STUDY AIMS: Endoscopic balloon sphincteroplasty (EBS) has been reported to be a safe alternative to sphincterotomy for the treatment of bile duct stones. We evaluated the factors which influence the therapeutic efficacy of EBS. PATIENTS AND METHODS: A total of 118 consecutive patients with bile duct stones were treated by EBS. After conventional endoscopic retrograde cholangiography (ERC), EBS was done using a biliary dilation catheter (balloon diameter, 8mm). The duct was then cleared using Dormia baskets or retrievel balloon catheters. When the stones were greater than 8 mm in diameter, mechanical lithotripsy was performed before extraction. Complete stone clearance was assessed by balloon-ERC and intraductal ultrasonography. Therapeutic efficacy was assessed using univariate and multivariate analysis. Patients were classified into three groups according to the bile duct diameter: nondilated (bile duct < or = 10 mm), mildly dilated (10 mm < bile duct < or = 15 mm), and severely dilated group (bile duct > 15 mm). RESULTS: In 113 of 118 (96%) patients, the stones were completely cleared with one to six endoscopic sessions (mean 1.6 sessions). In the nondilated group, 24 of 28 (85%) patients were cleared of stones in one session (mean 1.2 sessions), without the use of mechanical lithotripsy in 23 of 28 (82 %) patients. In the mildly dilated group, 23 of 38 (61 %) patients were cleared of stones in one session (mean 1.5 sessions). In contrast, in the severely dilated group, only 16 of 52 (31 %) patients were cleared of stones in one session (mean 2.0 sessions). Stone size, number of stones, and use of mechanical lithotripsy were independent variables which influenced the success of stone clearance in one session after EBS. CONCLUSION: When EBS is done in patients with bile duct stones, bile duct diameter may be a good indicator of therapeutic efficacy. In patients with severely dilated bile ducts (> 15 mm), EBS is of limited effectiveness.

Aged↗
PubMed1999

Fluorescence in situ hybridization using 16S rRNA-targeted oligonucleotides reveals localization of methanogens and selected uncultured bacteria in mesophilic and thermophilic sludge granules.

16S rRNA-targeted in situ hybridization combined with confocal laser scanning microscopy was used to elucidate the spatial distribution of microbes within two types of methanogenic granular sludge, mesophilic (35 degrees C) and thermophilic (55 degrees C), in upflow anaerobic sludge blanket reactors fed with sucrose-, acetate-, and propionate-based artificial wastewater. The spatial organization of the microbes was visualized in thin sections of the granules by using fluorescent oligonucleotide probes specific to several phylogenetic groups of microbes. In situ hybridization with archaeal- and bacterial-domain probes within granule sections clearly showed that both mesophilic and thermophilic granules had layered structures and that the outer layer harbored mainly bacterial cells while the inner layer consisted mainly of archaeal cells. Methanosaeta-, Methanobacterium-, Methanospirillum-, and Methanosarcina-like cells were detected with oligonucleotide probes specific for the different groups of methanogens, and they were found to be localized inside the granules, in both types of which dominant methanogens were members of the genus Methanosaeta. For specific detection of bacteria which were previously detected by whole-microbial-community 16S ribosomal DNA (rDNA)-cloning analysis (Y. Sekiguchi, Y. Kamagata, K. Syutsubo, A. Ohashi, H. Harada, and K. Nakamura, Microbiology 144:2655-2665, 1998) we designed probes specific for clonal 16S rDNAs related to unidentified green nonsulfur bacteria and clones related to Syntrophobacter species. The probe designed for the cluster closely related to Syntrophobacter species hybridized with coccoid cells in the inner layer of the mesophilic granule sections. The probe for the unidentified bacteria which were clustered with the green nonsulfur bacteria detected filamentous cells in the outermost layer of the thermophilic sludge granule sections. These results revealed the spatial organizations of methanogens and uncultivated bacteria and their in situ morphologies and metabolic functions in both mesophilic and thermophilic granular sludges.

Anaerobiosis↗
PubMed1999

Possible involvement of proteolytic degradation of tyrosinase in the regulatory effect of fatty acids on melanogenesis.

The purpose of this study was to investigate the mechanism of fatty acid-induced regulation of melanogenesis. An apparent regulatory effect on melanogenesis was observed when cultured B16F10 melanoma cells were incubated with fatty acids, i.e., linoleic acid (unsaturated, C18:2) decreased melanin synthesis while palmitic acid (saturated, C16:0) increased it. However, mRNA levels of the melanogenic enzymes, tyrosinase, tyrosinase-related protein 1 (TRP1), and tyrosinase-related protein 2 (TRP2), were not altered. Regarding protein levels of these enzymes, the amount of tyrosinase was decreased by linoleic acid and increased by palmitic acid, whereas the amounts of TRP1 and TRP2 did not change after incubation with fatty acids. Pulse-chase assay by [35S]methionine metabolic labeling revealed that neither linoleic acid nor palmitic acid altered the synthesis of tyrosinase. Further, it was shown that linoleic acid accelerated, while palmitic acid decelerated, the proteolytic degradation of tyrosinase. These results suggest that modification of proteolytic degradation of tyrosinase is involved in regulatory effects of fatty acids on melanogenesis in cultured melanoma cells.

Animals↗
PubMed1998

Expression of protooncogene c-kit and its ligand stem cell factor (SCF) in gastric carcinoma cell lines.

We examined 13 human gastric carcinoma cell lines for the expression of both c-kit and stem cell factor (SCF). Expression of mRNAs was detected by both Northern blot analysis and reverse transcriptase-polymerase chain reaction (RT-PCR), and expression of translated proteins was detected by western blotting. Using RT-PCR we confirmed the expression of c-kit in five (ECC12, TMK1, MKN7, GCIY, and HGC27) cell lines. Northern blot analysis showed coexpression of both c-kit and SCF in ECC12 and expression of SCF in five other (MKN74, MKN1 OKAJIMA, KATOIII, and TMK1) cell lines. SCF stimulated both tyrosine phosphorylation of c-kit and growth of ECC12, whereas it did not stimulate those of GCIY. The sizes of c-kit transcript and protein in GCIY were slightly smaller than those of the reported ones, suggesting the presence of a biologically inactive truncated form of c-kit in GCIY. The present study suggests that c-kit/SCF system might play an important role in the carcinogenesis and tumor growth of ECC12 and that the truncated form of c-kit in GCIY might not be associated with malignant transformation.

Blotting, Northern↗
PubMed1998

Immunoreactivity of alpha-melanocyte-stimulating hormone, adrenocorticotrophic hormone and beta-endorphin in cutaneous malignant melanoma and benign melanocytic naevi.

Melanocyte-stimulating hormone (MSH) has been reported to enhance the experimental metastatic behaviour of melanoma cells in the mouse model. alpha-MSH production and MSH receptor (melanocortin 1 receptor gene) expression have been detected in cultured normal human melanocytes and metastasized melanomas. The exact role of MSH in the metastatic behaviour of human melanoma cells is, however, not yet known. To clarify a possible role of proopiomelanocortin (POMC)-derived peptides, including alpha-MSH, in melanoma development and progression, we analysed immunohistochemically the localization of alpha-MSH adrenocorticotrophic hormone (ACTH) and beta-endorphin in various kinds of benign pigmented naevocytic lesions and malignant melanomas. Three of 21 samples of common and dysplastic naevi showed detectable alpha-MSH staining in naevus cells, and five and six of 15 samples were weakly positive for ACTH and beta-endorphin staining, respectively. In melanoma samples, 24 of 45, 23 of 39 and 30 of 42 samples showed positive staining with alpha-MSH, ACTH and beta-endorphin antibodies, respectively. Furthermore, staining for all three antibodies was noted to be more intense and diffuse in samples of nodular melanoma, vertically growing acral lentiginous melanoma and superficial spreading melanoma as well as metastatic lesions compared with those of naevi. Although it is yet to be determined whether or not this strong staining for POMC-derived peptides in advanced melanoma cells indicates a role of autocrine or paracrine regulation, our results suggest a possible involvement of POMC gene products in melanoma progression.

Adrenocorticotropic Hormone↗
PubMed1998

Modulation of melanocyte-stimulating hormone receptor expression on normal human melanocytes: evidence for a regulatory role of ultraviolet B, interleukin-1alpha, interleukin-1beta, endothelin-1 and tumour necrosis factor-alpha.

Melanocyte-stimulating hormone (MSH) receptor binding activity and melanocortin-1 receptor (MC1-R) gene expression on normal human melanocytes have been studied as responses to the effects of ultraviolet B (UVB), interleukin-1 (IL-1), endothelin-1 (ET-1) and tumour necrosis factor-alpha (TNF-alpha), which are known as UV sensitive regulators of melanocytic function. MSH receptor (MSH-R) binding activity was upregulated by UVB, IL-1alpha, -1beta and ET-1, but was downregulated by TNF-alpha. Northern blot analysis showed that MC1-R mRNA expression was induced 24 h after UVB irradiation in a dose-dependent manner, and that 24-h treatment with ET-1 also induced an expression of MC1-R mRNA, whereas TNF-alpha downregulated the expression. In addition, IL-1alpha and -1beta have a small but real inductive effect on MC1-R mRNA expression. Taken together, our results suggest a model in which higher MC1-R mRNA expression is accompanied by upregulation of MSH-R binding activity, and enhanced by UVB or cytokines sensitive to UVB. Such a regulatory system would enable normal human melanocytes to respond to MSH more efficiently and induce an increase of melanization of the skin through the MSH/MSH-R system after UVB radiation.

Adolescent↗
PubMed1997

BDNF and NT-3 modulate expression and threonine phosphorylation of microtubule-associated protein 2 analogues, and alter their distribution in the developing rat cerebral cortex.

Effects of brain-derived neurotrophic factor (BDNF) and neurotrophin (NT)-3 on the expression of structural or synapse-associated proteins were examined in the developing rat cerebral cortex. Following ventricular administration of BDNF or NT-3 at embryonic day (E) 16, expression of microtubule-associated protein (MAP) 2 of 280 kDa was enhanced at E18 and/or E20, and threonine phosphorylation of MAP2 analogues of 120 and 66 kDa was modulated in different ways. NT-3 basically altered the distribution of MAP2 proteins at E20. These findings suggest that NT-3 and BDNF play a role in regulating production and phosphorylation of MAP2 analogues during development of the rat cerebral cortex.

Animals↗