Biomedical subjects

T Tsuji

Publications and source records attributed to T Tsuji.

At least 487 records · Page 27Linked to original sources

PubMed1996

Tenascin expression in endomyocardial biopsy specimens in patients with dilated cardiomyopathy: distribution along margin of fibrotic lesions.

OBJECTIVE: To examine the hypothesis that tenascin, an extracellular matrix glycoprotein, contributes to fibrotic changes in dilated cardiomyopathy. METHODS: The localisation of tenascin in biopsy specimens of the hearts obtained from eight patients with dilated cardiomyopathy was examined using staining by the avidin-biotin-peroxidase complex method. RESULTS: (1) Perimysium and endomysium. Although positive staining for tenascin was observed in the enlarged perimysium and endomysium in all patients, moderately intense staining was characteristically observed near the replacement fibrotic lesions. In the narrow perimysium and endomysium of the myocardium not containing replacement fibrotic lesions, tenascin was not present, as in the control specimens. (2) Replacement fibrotic lesions. Non-homogeneous positive staining for tenascin was detected in all replacement fibrotic lesions examined. Intense tenascin deposition was observed in the peripheral portion of the replacement fibrotic lesions. The tenascin staining observed in the small replacement fibrotic lesions was more intense than that in the large lesions. CONCLUSIONS: Tenascin contributes to the development of the fibrotic changes seen in the dilated cardiomyopathic heart. Its characteristic location, specifically the distribution along the margin of the fibrosis, suggests that fibrotic change is a continuous process in hearts with dilated cardiomyopathy.

Adult↗

PubMed1996

The amino acids of Escherichia coli enterotoxin B subunit involved in binding to Bio-Gel A-5m or to the glycoprotein from mouse intestinal epithelial cells.

We determined whether Arg13, Met31, and Ser95 of the heat-labile enterotoxin B subunit (LT-B) might be involved in Lt-B binding to oligosaccharides, which did not bind to the B subunit of the cholera toxin (CT-B). Three LT-B mutants, R13H, M31L, and S95A were prepared by substituting three amino acid residues that differ in CT-B. These mutants formed a pentamer and exhibited the same binding ability to the GM1 ganglioside as native LT-B. Although these mutants did not bind to Bio-Gel A-5m, they did bind to the glycoprotein from mouse intestinal cells in the order R13H > M31L > S95A. These data suggest that Ser95, Met31, and Arg13 are important for LT-B binding to Bio-Gel A-5m, and that although Ser95 is also partially responsible for LT-B binding to the glycoprotein, Arg13 has no significant involvement in it.

Animals↗

PubMed1996

Salivary EGF regulates eosinophil-derived TGF-alpha expression in hamster oral wounds.

Using hamster as an oral wound healing model, we examined eosinophils and their expression of transforming growth factor-alpha (TGF-alpha) and transforming growth factor-beta 1 (TGF-beta 1). Oral wounds healed approximately two times faster than their cutaneous counterparts. Eosinophils infiltrated prominently into oral wounds; however, unlike the dual expression of TGF-alpha and TGF-beta 1 in skin wounds, oral wound-associated eosinophils expressed TGF-beta 1, but not TGF-alpha. Because saliva is present in oral environments and contains epidermal growth factor (EGF) and TGF-alpha, sialoadenectomy was performed in this model to determine whether the lack of TGF-alpha expression by eosinophils in oral wounds is due to the presence of salivary EGF and/or TGF-alpha. We found that eosinophils in sialoadenectomized hamsters did express TGF-alpha during oral wound healing but that such expression was suppressed when EGF was added to their drinking water. Taken together, our findings suggest that eosinophil-derived TGF-alpha and salivary TGF-alpha/ EGF may have complementary roles in contributing to TGF-alpha in oral wound healing.

Animals↗

PubMed1996

Molecular biology of human oral cancer.

The application of molecular biological tools to the study of cancer has significantly advanced the field of human cancer research. Such study has demonstrated the involvement of two classes of highly conserved cellular genes in the malignant transformation process: oncogenes and tumor suppressor genes. Despite these advances in the molecular biology of human cancers, our understanding of human oral cancer lags behind that of cancer of other body sites. This review attempts to assess the current status of the molecular biology of human oral cancer.

Cell Adhesion Molecules↗

PubMed1996

Concanamycin B, a vacuolar H(+)-ATPase specific inhibitor suppresses bone resorption in vitro.

Effects of concanamycin B, a specific inhibitor of the vacuolar type H(+)-ATPase (V-ATPase), on the stimulation of bone resorption induced by parathyroid hormone (PTH) were examined in vitro. Concanamycin B was found to inhibit PTH-stimulated osteoclastic pit formation and to suppress the acidification of vacuolar organelles by V-ATPase in the osteoclasts. PTH-stimulated 45Ca release from prelabelled chick embryonic calvariae was also inhibited by concanamycin B in a dose-dependent manner. These results suggest that osteoclastic acidification of lacunae by V-ATPase plays an essential role in mineral dissolution and degradation of the organic matrix during bone resorption.

Animals↗

PubMed1996

[Reaction of 3-hydroxy-dihydrosantonin with TsOH. A formal synthesis of dihydrocotunolide].

In connection with our another project a relatively large quantity of 5 was required. We, therefore, investigated the reaction of 3 with TsOH; this gave a diene mixture 7a, 7b, 6 and 5 in a ratio of 8:2:4:1. Almost the same 8:2:1 "equilibrium mixture" of the dienes 7a, 7b and 5 was also obtained when the purified samples of 5 and 7a were independently treated with TsOH under the same conditions. The above results suggest that the dienes might be produced in the following way; beta-elimination of H2O from 3 gives a homoannular diene 5. The protonation to the more electron-rich tetrasubstituted C4-C5 double bond of 5 from the less hindered alpha-side followed by the isomerization of the double bond gives 7a. 7b is formed via 7a and/or 5 by the double bond isomerizations. On the other hand, 6 is formed by concerted elimination of H2O between C3-OH and C6-H. As a result of these studies, the synthetic precursor of dihydrocostunolide 7b which was already prepared by Corey in 6 steps (ca. 7%) from santonin was synthesized in 3 steps (ca. 6%) from the latter.

Antimalarials↗

PubMed1996

Sequential changes in laminin and type IV collagen in the infarct zone--immunohistochemical study in rat myocardial infarction.

BACKGROUND: The healing process, which affects ventricular remodeling, is an important factor in the prognosis of myocardial infarction. We hypothesized that laminin and type IV collagen contribute to extracellular matrix assembly in healing after myocardial infarction. We examined sequential changes in these two components after experimental myocardial infarction in rats. MATERIALS AND METHODS: Hearts were excised from 1 day to 10 weeks after permanent left coronary ligation in rats. Immunohistochemical staining with a polyclonal antibody to laminin and type IV collagen was performed by the avidin-biotin-peroxidase method. RESULTS: Laminin: On day 3, laminin initially appeared in a wavy fashion in the granulation tissue of the infarct peripheral zone and was not restricted to the cell membrane; the staining distribution in the peripheral zone then gradually increased, reaching a maximum on days 7-11. The distribution progressed from the peripheral zone to the outer lesion of the central zone of the infarct for 1-2 days, and reached the center point after 2 weeks. The extent of the staining distribution gradually decreased after reaching this maximum, but the staining did not completely disappear. Type IV Collagen: Changes in type IV collagen were essentially the same as those in laminin. A wavy staining pattern of type IV collagen appeared in the infarct peripheral zone from day 3, reached its maximum extent on days 7-11, and decreased gradually thereafter. The distribution progressed from the peripheral zone to the outer lesion of the central zone for 1-2 days, reaching the center point after 2 weeks. CONCLUSIONS: Laminin and type IV collagen contribute to extracellular matrix formation in the infarct zone relatively early after myocardial infarction.

Animals↗

PubMed1996

Cyclooxygenase induction is essential for NGF synthesis enhancement by NGF inducers in L-M cells.

Nerve growth factor (NGF) inducers, for example, 4-methylcatechol, pyrroloquinoline quinone, kansuinin A, and ingenol triacetate, stimulate NGF synthesis in L-M cells, but the mechanism of NGF induction by NGF inducers is not known. Using the four different types of previously described NGF inducers, we proved induction of cyclooxygenase activity by NGF inducers and detected prostaglandins D2 and E2 as metabolites of arachidonic acid. From the observation that the induction of NGF by each NGF inducers was inhibited by cyclooxygenase inhibitors or dexamethasone, cyclooxygenase activation is supposed to be an essential process for NGF induction.

Animals↗

PubMed1996

Extracellular matrix components in dilated cardiomyopathy. Immunohistochemical study of endomyocardial biopsy specimens.

Fibrotic change is one of the characteristic features of the heart with idiopathic dilated cardiomyopathy (DCM), but the extracellular matrix components have not been fully clarified. Fibronectin, laminin, and types IV, I and III collagen staining using the avidin-biotin-peroxidase complex method was conducted to study the localization of extracellular matrix components in biopsy specimens obtained from 7 patients with DCM. Fibronectin was observed in the endomysium and perimysium, co-existent with types I and III collagen; it was also distributed nonhomogeneously in the replacement fibrotic lesions. The staining intensity in a section varied substantially from location to location. Laminin surrounded the myocytes and was distributed in the endomysium localized together with type IV collagen, but it was not found in the perimysium. Weak but positive staining was observed in replacement fibrotic lesions in 2 patients. Similarly to laminin, type IV collagen staining was observed around the myocytes and in the endomysium. It was also observed in replacement fibrotic lesions in 4 patients either with or without laminin. Type I collagen, localized together with type III collagen, was distributed in both the endomysium and perimysium. It was also distributed nonhomogeneously in the replacement fibrotic lesions. Similar staining to type I collagen was seen for type III collagen. In summary, all examined components were observed in the fibrotic lesions with no primary deficit of any examined component being demonstrated. The variability of staining intensity and positive fibronectin staining suggests that the phases of fibrotic changes differed substantially from location to location.

Adolescent↗

PubMed1996

Cathepsin B in the growth of colorectal cancer: suppressive effect of leupeptin on the growth of DMH-induced rat colon neoplasm.

Cathepsin B, a thiol protease, has been reported to be involved in cancer progression and metastasis. The suppressive effects of two kinds of protease inhibitors, leupeptin and dietary camostate (FOY-305), on tumorigenesis and progression in 1, 2-dimethylhydrazine (DMH)-induced rat colon neoplasm were examined in relation to tissue cathepsin B activity. Male Donryu rats were treated with leupeptin or FOY-305 during or after the administration of DMH. There were no significant differences in average tumor numbers among all DMH-treated groups. However, the percentage of small tumors was significantly higher in the group in which leupeptin was supplied during DMH administration. This trend was not recognized in the FOY-305-treated groups. The ratio of cathepsin B activity in the tumors to that in the tumor-bearing tissue (T/Tb) was significantly increased with increasing tumor size (P = 0.009). The cathepsin B activity levels in the tumor-bearing mucosa in the groups which received leupeptin or FOY-305 following DMH treatment were both significantly lower than that in the group which received neither protease inhibitor (P = 0.046 and P = 0.0067, respectively). The results obtained indicate that leupeptin may have suppressed tumor growth by lowering the tissue cathepsin B activity.

1,2-Dimethylhydrazine↗

PubMed1996

Cathepsin B in the growth of colorectal cancer: increased activity of cathepsin B in human colorectal cancer.

Cathepsin B, a thiol protease, is involved in cancer metastasis. To clarify the role of cathepsin B in tumor progression in human colorectal cancer, the relationship between its activity, immunohistochemical staining, and clinical tumor progression was investigated. Cathepsin B activity in adenocarcinomas was significantly elevated compared with that in the tumor-bearing tissue. Furthermore, the tumor/tumor-bearing tissue (T/Tb) ratio of the activity was significantly higher than that of colorectal adenoma. Immunohistochemical studies demonstrated intense staining in the cancerous tissue. With respect to the clinical stage of tumors, the activity tended to be higher in tumors that had invaded the serosa or subserosa than in those that invaded the proper muscle. The results suggest that cathepsin B participates in the progression of human colorectal cancer, and its increased expression is a sensitive marker of the differentiation between colorectal adenoma and adenocarcinoma.

Adenocarcinoma↗

PubMed1996

Spheroid cultures of human hepatoblastoma cells (HuH-6 line) and their application for cytotoxicity assay of alcohols.

Spheroid cultures of human hepatoblastoma cells (HuH-6 line) were established by rotating 3 x 10(6) cells/3 ml culture medium in 25-ml Erlenmeyer flasks on a gyratory shaker. The size of the spheroids rapidly increased until 4 days of culture, and thereafter their size gradually increased until 8 days of culture. A considerable amount of lactate dehydrogenase (LDH) was detected in the culture medium at 24h after seeding because of cell damage by subculturing, but thereafter the amount released was small, indicating that the spheroids were in healthy condition. Albumin production, one of the differentiated functions of hepatocytes, was higher in spheroid cultures than in monolayer cultures. Using this spheroid culture model, the cytotoxic effects of alcohols on HuH-6 cells were studied by measuring the activity of LDH released in the medium from damaged cells. The results indicate that the increasing order of toxicity of the alcohols was as follows: methanol < ethanol < propanol.

Albumins↗

PubMed1996

Usefulness of indocyanine green injection during ultrasound-guided liver biopsy for the diagnosis of small hepatocellular carcinoma.

To diagnose hepatocellular carcinoma (HCC) functionally and immediately, we examined the usefulness of indocyanine green (ICG) injection during ultrasound-guided liver biopsy. Liver specimens were obtained after intravenous ICG injection by ultrasound-guided biopsy from 251 space-occupying lesions (SOL) in 136 patients. The tissues were immediately examined for ICG uptake using an infrared Vidicon camera and were also subjected to histopathological examinations. Of the 112 ICG-negative biopsy specimens, 105 were histologically diagnosed as HCC, 6 as dysplastic nodules (DN) and 1 as a regenerative nodule (RN). Of the 139 ICG-positive specimens, 18 were diagnosed as HCC, 1 as DN and 120 as RN. The sensitivity of the absence of ICG uptake (SEAIU), the specificity of the absence of ICG uptake (SPAIU), and the positive predictive value of the absence of ICG uptake (PPAIU) for the diagnosis of HCC were 85.3%, 94.5% and 93.8%, respectively. Of the 251 SOLs, 184 were less than 2 cm. SEAIU, SPAIU and PPAIU for the diagnosis of these small HCC were 85.3%, 94.5% and 91.4%, respectively. These results support the reliability of ICG injection during ultrasound-guided liver biopsy to diagnose even small HCC.

Aged↗

PubMed1996

Primary sclerosing cholangitis in Japanese patients: association with inflammatory bowel disease.

To characterize primary sclerosing cholangitis (PSC) in Japanese patients and its association with inflammatory bowel disease (IBD), 155 reported cases of PSC, including 6 cases of our own, were reviewed. The prevalence of IBD was less in Japanese PSC patients than in Western patients (23% versus 62-100%). Japanese PSC patients with IBD were younger (mean age, 33.1 versus 51.8 years) and were more often women (51% versus 36%) than those without IBD. Seventy-four percent of PSC patients with IBD had extensive colonic lesions, and 89% of those developed IBD simultaneously, with or prior to PSC. There were 3 cases of neutrophilic cholangitis among the PSC patients with IBD but none in those without IBD. Based on these observations, we speculate that there may be subtypes of PSC which differ pathophysiologically.

Adolescent↗

PubMed1996

Comparison of various methods of assaying the cytotoxic effects of ethanol on human hepatoblastoma cells (HUH-6 line).

The sensitivity of five kinds of cytotoxicity assays using ethanol on human hepatoblastoma cells (HUH-6 line), which were cultured as monolayers or spheroids, was compared. Ethanol was chosen as a test because it acts on cell membranes directly without being metabolized and exerts its cytotoxicity. The assay methods used were as follows: 3- (4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT), lactate dehydrogenase (LDH), colony formation, cell growth and DNA assays. The sensitivity of the assays was: LDH < DNA < cell growth < MTT < colony formation. LDH assay had the advantage that the same culture could be used for multiple assays, but when a small number of cells were assayed, no significant increase in the release of LDH was detected in the assay cultures compared with the control cultures. Although the DNA and cell growth assays were more sensitive than the LDH assay, the extent of cell damage may be underestimated because the damaged cells and DNA present in the cultures are included in the assay samples. On the other hand, both MTT and colony formation assays showed a high sensitivity. The MTT assay was done within 24 h after ethanol was added to the cultures and was applicable to both monolayer and spheroid cultures, while the colony formation assay required 1-2 weeks and it was applicable only to monolayer cultures. Taken together, the MTT assay was the most suitable method to evaluate the cytotoxic effects of ethanol on HUH-6 cells cultured as either monolayers or spheroids.

Cell Survival↗

PubMed1996

Purification and characterization of a 39kDa apurinic/apyrimidinic endonuclease from mouse ascites sarcoma cells.

We purified an apurinic/apyrimidinic (AP) endonuclease from mouse ascites sarcoma (SR-C3H/He) cells. The enzyme showed nicking activity on acid-depurinated DNA but not on untreated, intact DNA. It also showed priming activity for DNA polymerase on both acid-depurinated and bleomycin-damaged DNA. The priming activity on bleomycin-damaged DNA was two times higher than that on an acid-depurinated DNA. The enzymatic properties indicate that the enzyme is a class II AP endonuclease having DNA 3' repair diesterase activity. The purified enzyme has a molecular weight of 39,000 as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The optimal pH for AP endonuclease activity was 8.0 in 50 mM Tris-HCl buffer. The AP endonuclease activity depended on divalent cation such as Mg2+ and Co2+ ions, and was inhibited by 2 mM EDTA with no addition of the divalent cation. An appropriate concentration of sodium or potassium salt stimulated the activity. Partial digestion of the AP endonuclease with Staphylococcus aureus V8 protease produced 4 major peptide fragments which may be used for protein sequencing.

Acids↗

PubMed1996

Relationship between response to interferon therapy and detection of hepatitis C virus RNA by differential flotation centrifugation.

Circulating hepatitis C virus (HCV) particles can be fractionated by means of differential flotation centrifugation. It is reported that in the bottom fraction HCV is in the form immune complexes, whereas in the top, it is free of antibodies. We evaluated the significance of circulating complex and free HCV in chronic hepatitis C, and assessed the relationship in terms of the response to interferon (IFN) therapy. We examined sera before, just after, and 1 year after administering IFN to 18 patients with chronic hepatitis C, 10 of whom responded (group CR), and 8 did not (group NR). The amounts of virus were similar between both groups before therapy. After differential flotation centrifugation with 1.063 g/ml of NaCl, the top and bottom fractions were assayed for HCV RNA. Before therapy, HCV RNA was detected in the top fraction in 1 of 10 in group CR, and in 6 of 8 in group NR (P < 0.05, chi-square test). HCV RNA was positive in the bottom fraction of all samples. In a follow-up study of group NR, HCV RNA was detected in the top fraction in 3 of 8 just after IFN therapy, and in 7 of 8 after 1 year. This study suggests that the presence of HCV in the top fraction can predict a poor response to IFN therapy.

Adult↗

PubMed1996

Meningeal sinus histiocytosis mimicking lymphoplasmacyte-rich meningioma. Case report.

Extranodal sinus histiocytosis with massive lymphadenopathy involving the meninges is a rare clinical entity. The authors describe the unusual case of an occipital meningeal lesion in a 25-year-old man in whom they found inflammation of the occipital lobe mimicking lymphoplasmacyte-rich meningioma. The clinical similarities and differences are discussed.

Adult↗