Biomedical subjects

T Fujimura

Publications and source records attributed to T Fujimura.

At least 91 records · Page 5Linked to original sources

PubMed1999

Live Bartonella henselae enhances endothelial cell proliferation without direct contact.

The proliferation of human umbilical vein endothelial cells (HUVECs) cocultivated with live B. henselae was enhanced in a bacterial dose-dependent manner, and the stimulatory effect was specific to vascular endothelial cells. The inactivation of B. henselae by UV or heat treatment abolished its stimulatory activity, suggesting that live bacteria is necessary for the growth stimulation effect. To investigate the role of direct contact, live B. henselae were separated from HUVECs by a filter membrane (Millicell-CM insert). Even under this condition, an enhanced proliferation of HUVECs was observed. However, no morphological changes in the HUVECs were apparent compared to the B. henselae -infected cells. Furthermore, we isolated a nonpiliated strain of B. henselae that is unable to attach to and enter into endothelial cells. The nonpiliated strain possessed the ability to stimulate the proliferation of cocultivated HUVECs the same as the piliated strain. Moreover, the culture supernatants of B. henselae were also able to induce HUVEC proliferation. Our results indicate that the stimulation of HUVEC proliferation by B. henselae is mediated by soluble factor(s) secreted from the bacteria.

Angiomatosis, Bacillary↗

PubMed1999

Intraoperative lavage for cytological examination in 1,297 patients with gastric carcinoma.

BACKGROUND: This study examined the clinical value of intraoperative peritoneal lavage for cytological examination in patients with gastric cancer. Peritoneal dissemination is the most frequent mode of recurrence for this tumor. METHODS: A retrospective of lavage findings, other factors, and outcome was performed in 1,297 patients with gastric cancer who underwent intraoperative peritoneal lavage. RESULTS: The 5-year survival rate of patients with positive lavage cytology was only 2%. Patients who underwent curative resection and had negative cytology had a significantly better 5-year survival rate (P < 0.001). Even among patients with macroscopic peritoneal dissemination, the survival rate was significantly better with negative cytology, which reflected fewer free cancer cells in the peritoneal cavity. Serum concentrations of carcinoembryonic antigen and carbohydrate antigen 19-9 were significantly higher in patients with positive cytology. Multivariate analyses indicated that intraoperative cytological findings was an independent prognostic factor for survival, and was the most important factor for predicting peritoneal recurrence. CONCLUSIONS: Intraoperative peritoneal lavage cytology is important in predicting survival and peritoneal recurrence in gastric cancer.

Biomarkers, Tumor↗

PubMed1999

Immunoreactivity of endothelins and endothelin receptor in the stria vascularis of the mouse cochlea.

Immunoreactivities of endothelin-1, endothelin-3, endothelin receptor type A, and Na,K-ATPase were investigated in the stria vascularis of adult male WBB6F1 +/+ mice and in that of W/Wv mutants lacking strial intermediate cells. In the +/+ mice, electron microscopic immunoreactivity for the endothelins was seen on the rough endoplasmic reticulum, Golgi apparatus, cytoplasmic vesicles and lysosomes exclusively in the strial intermediate cells by the postembedment method. Immunoreactive endothelin receptor A was localized along the plasma membrane of strial marginal cells of both wild and mutant types although the immunoreactivity of the latter was much less than that of the former by the preembedment method. These findings suggest that the endothelins, which are produced in the strial intermediate cells, may play a role in the maintenance of the stria vascularis function in the +/+ mice. Since the plasma membrane of the marginal cells of the W/Wv mice, which do not generate a high positive endocochlear potential, also showed immunoreactivity for Na,K-ATPase, it seems likely that the endothelins are involved in the activation of sodium pump of the strial marginal cells by mediation of endothelin receptor A. In addition, the role of lysosomes in the crinophagy of the endothelins in the strial intermediate cells is proposed in the +/+ mice.

Animals↗

PubMed1999

Interaction between surface active drug (FK906:rennin inhibitor) and cyclodextrins in aqueous solution.

The aggregation behavior of FK906, which is a peptide like hypertensive agent, in aqueous solution was studied by static light scattering, 1H-nuclear magnetic resonance (NMR), and surface tension. These experiments showed a clear critical micelle concentration (cmc) at around 6.3 x 10(-3) to 1.3 x 10(-2) M of FK906 aqueous solution. The result of 1H NMR experiments revealed that FK906 aggregates primarily by hydrophobic interactions involving the benzyl moiety. The Debye plots from light-scattering studies showed that the apparent molecular weight of aggregated FK906 molecule is 1670 which corresponds to 2-3 molecules of FK906. The effect of alpha- and beta-cyclodextrins on the surface tension of FK906 aqueous solution was investigated. It appeared that the addition of alpha-cyclodextrin showed very small shift of cmc, but that of beta-cyclodextrin shifted the cmc to much higher concentration. The investigation on the surface tension of FK906 aqueous solution in the presence of beta-cyclodextrin indicated that FK906 forms a 1:1 complex with beta-cyclodextrin. On the basis of these experiments, it appears that beta-cyclodextrin has an ability to change the surface active property of FK906 in its aqueous solution. Therefore, it is expected that the addition of beta-cyclodextrin to FK906 aqueous solution may prevent the adsorption onto container walls and/or reduce the local irritancy.

Antihypertensive Agents↗

PubMed1999

Chemical induction of disease resistance in rice is correlated with the expression of a gene encoding a nucleotide binding site and leucine-rich repeats.

Probenazole (3-allyloxy-1,2-benzisothiazole-1,1-dioxide) is an agricultural chemical primarily used to prevent rice blast disease. Probenazole-treated rice acquires resistance to blast fungus irrespective of the rice variety. The chemical is applied prophylactically, and is thought to induce or bolster endogenous plant defenses. However, the mechanisms underlying this effect have not been established. To understand the mode of the chemical's action, we screened for novel probenazole-responsive genes in rice by means of differential display and identified a candidate gene, RPR1. RPR1 contains a nucleotide binding site and leucine-rich repeats, thus sharing structural similarity with known disease resistance genes. The expression of RPR1 in rice can be up-regulated by treatment with chemical inducers of systemic acquired resistance (SAR) and by inoculation with pathogens. RPR1-related sequences in rice varieties seem to be varied in sequence and/or expression, indicating that RPR1 itself is not a crucial factor for induced resistance in rice. However, Southern blot analysis revealed the existence of homologous sequences in all varieties examined. While the role of RPR1 has yet to be clarified, this is the first report of the identification of a member of this gene class and its induction during the systemic expression of induced disease resistance.

Amino Acid Sequence↗

PubMed1999

All-trans retinoic acid promotes the repair of tortuosity of elastic fibres in rat skin.

To elucidate the repair effects of all-trans retinoic acid (t-RA) on ultraviolet (UV)-induced tortuosity of elastic fibres in rat skin, the hind limbs of Sprague-Dawley rats were irradiated at a suberythemal dose of UV (three times/week for 6 weeks) and 0.025% t-RA in ethanol was applied topically five times/week for 6 weeks. The three-dimensional arrangement of elastic fibres with special reference to their linearity was quantified by image analysis using a scanning electron microscope following a combination of intravascular resin injection and selective digestion using formic acid. When t-RA was topically applied for 6 weeks on wrinkles formed by 6 weeks' exposure of the skin of rat hind limbs to UV radiation, the wrinkles disappeared to control levels, concomitant with an improvement in skin elasticity. The linearity of the elastic fibres was significantly (P < 0.01) increased compared with age-matched UV-irradiated controls. These findings suggest that the degeneration and deposition of elastic fibres accompanied by an increase in their linearity is involved in the process of wrinkle repair by topical application of t-RA.

Administration, Topical↗

PubMed1999

Purification and characterization of caffeine synthase from tea leaves.

Caffeine synthase (CS), the S-adenosylmethionine-dependent N-methyltransferase involved in the last two steps of caffeine biosynthesis, was extracted from young tea (Camellia sinensis) leaves; the CS was purified 520-fold to apparent homogeneity and a final specific activity of 5.7 nkat mg-1 protein by ammonium sulfate fractionation and hydroxyapatite, anion-exchange, adenosine-agarose, and gel-filtration chromatography. The native enzyme was monomeric with an apparent molecular mass of 61 kD as estimated by gel-filtration chromatography and 41 kD as analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The enzyme displayed a sharp pH optimum of 8.5. The final preparation exhibited 3- and 1-N-methyltransferase activity with a broad substrate specificity, showing high activity toward paraxanthine, 7-methylxanthine, and theobromine and low activity with 3-methylxanthine and 1-methylxanthine. However, the enzyme had no 7-N-methyltransferase activity toward xanthosine and xanthosine 5'-monophosphate. The Km values of CS for paraxanthine, theobromine, 7-methylxanthine, and S-adenosylmethionine were 24, 186, 344, and 21 microM, respectively. The possible role and regulation of CS in purine alkaloid biosynthesis in tea leaves are discussed. The 20-amino acid N-terminal sequence for CS showed little homology with other methyltransferases.

Affinity Labels↗

PubMed1999

Characterization of infiltrating T cells in human scalp explants from alopecia areata to SCID nude mice: possible role of the disappearance of CD8+ T lymphocytes in the process of hair regrowth.

T cells may play a role in the pathogenesis of alopecia areata (AA). We attempted to elucidate the linkage between infiltrating T cells and hair regrowth processes by grafting scalp skin from the affected region of patients with AA onto severe combined immune deficiency (SCID) nude mice. When the AA scalp was grafted into the mice, the grafts were accepted, and normal hair regrowth was observed. Before grafting, CD4+ and CD8+ T cells had infiltrated into the peribulb area. After grafting, the telogen hair shifted to anagen hair, and the CD4+ and CD8+ T cell infiltrates in the bulb area decreased in all cases. CD8+ T cells had almost disappeared from all portions of the follicles. It has been suggested that CD8+ T cells play a crucial role in the pathogenesis of AA. The absence of CD8+ T lymphocytes that responded to follicular autoantigens may induce hair regrowth in the grafted skin. In addition, the CD4+ human T cells that had infiltrated or still remained in the upper-middle portions including the bulge area accompanied the HLA-DR expression after grafting. Infiltrating or surviving T cell phenotypes and locations changed during the hair cycle in the grafts. These results indicate that the location of infiltrated T cells and their phenotypes may participate not only in hair loss but also in regrowth of hair in AA.

Adult↗

PubMed1999

Merits of soft nitriding scalers.

BACKGROUND: It has been demonstrated that nitriding modifies the physical characteristics of metals. The purpose of this study was to evaluate the changes induced by 3 levels of inexpensive soft nitriding treatments on commercial sickle scalers. METHODS: Taglite scalers (NT) were soft nitrided for 30 (SN30), 60 (SN60), or 90 (SN90) minutes. The cumulated scaled material was weighed every 10 strokes x 10 and thereafter every 1,000 to 8,000 strokes by an automatic scaling apparatus against epoxy resin. Weight differences were used to indicate abrasion resistance; the relative efficiency (RE) was calculated as the ratio of scaled amount at a given number of strokes (SN/NT). The hardness and the tensile strengths were determined for each soft nitriding treatment level. The nitrided layer thickness of representative SN scalers was observed by electron probe x-ray microanalysis. RESULTS: The SN60 was not significantly different from SN30 or SN90, but the SN90 was more efficient than SN30 for the first 100 strokes (P<0.01). The RE of all SN scalers was significantly greater (2.3 to 2.7 times) than the NT scalers from the beginning of the study and throughout all time periods (SN90 > SN60 > SN30); it increased further during the first 100 strokes (9.7 to 15.5 times), indicating the NT scalers wore out faster than SN scalers. The untreated scalers' performance decreased to 10% of baseline after 100 strokes; but even after 1,000 strokes, the SN60 and SN90 performed better than new untreated scalers. Thereafter, all scalers' performance, including SN scalers, decreased. While the NT blades ceased to cut measurable amounts after 7,000 to 8,000 strokes, all SN scalers continued to cut. Although SN90 scalers had the thickest soft layer and were the hardest (P<0.01), the SN60 had the highest tensile strength (P <0.01), suggesting that it might be the safest in practice. CONCLUSIONS: Sixty minutes of soft nitriding treatment of commercially available taglite scalers seem to be the optimal treatment duration to increase their durability, on the order of 100 to 1,000 times, without jeopardizing safety for clinical use.

Dental Alloys↗

PubMed1999

[A case report: modified PMUE therapy might be effective for advanced gastric carcinoma with pulmonary carcinomatous lymphangitis].

A regimen featuring a combination of CDDP, MMC, 5-FU and ETP (Modified-PMUE) was given to a patient with advanced gastric cancer containing pulmonary carcinomatous lymphangitis. This procedure made his carcinomatous lymphangitis disappear and primary and metastatic lesions reduce in size. The subsequent palliative reduction surgery for gastric carcinoma, proximal gastrectomy was feasible. Nevertheless, postoperative Modified-PMUE was ineffective: carcinomatous lymphangitis reappeared and progressed to death of respiratory failure postoperative 68 days. The SDI test, a sensitivity test for carcinostatic agent showed neither CDDP, MMC, 5-FU nor ETP sensitive to resected carcinoma cells. Therefore, we concluded that the Modified PMUE kills carcinoma cells sensitive to the agents and allows carcinoma cells resistant to the agents to grow.

Adenocarcinoma↗

PubMed1999

[The case report of minimal access CABG for triple-vessel disease].

A 55-year-old male patient underwent CABG for triple-vessel disease using the minimal access approach. The procedure was performed through a limited (10 cm) left para-sternal thoracotomy using extracorporeal circulation established with a usual aortic cannula, and pulmonary arterial and right atrial drainage. The myocardium was protected by antegrade administration of cold cardioplegic solution while the aorta was being cross-clampled. The saphenous vein graft was connected sequentially to the 4 PD and OM branches, and the left internal thoracic artery was grafted to the LAD. The postoperative course was uneventful and coronary angiography showed that all three grafts were patent. The patient was discharged one week postoperatively.

Coronary Artery Bypass↗

PubMed1999

[Surgery for metastatic brain and spinal tumor from colorectal cancer].

UNLABELLED: The results of invasive treatment of brain metastasis of colorectal cancer (6 cases) and spinal metastasis of colorectal cancer (3 cases), aimed at alleviating the symptoms, were analyzed. In all 6 cases of brain metastasis, metastases were also found in some other remote sites (often the liver). The average period from resection of the primary lesion to resection of the brain metastasis was 3 years and 3 months for this group. Of these 6 cases, 5 had cerebral metastasis and 1 had cerebellar metastasis. The preoperative performance status (PS) was 1 in 1 case and 2 in 5 cases. After surgery, the PS improved in all 6 cases, allowing all the patients to be discharged. There was only one patient whose death was attributable to brain metastasis. In all 3 cases of spinal metastasis, metastases were also detected in the lungs or liver (synchronous in) 1 case, metachronous in 2 cases). The preoperative PS was 2 in 1 case, 3 in 1 case and 4 in 1 case. After surgery, the PS improved in 2 cases. One of the three patients is still alive. CONCLUSION: The results of this study suggest that it is not impossible to improve the QOL of patients with brain or spinal metastasis of cancer by surgical intervention, although the response may vary from patient to patient.

Brain Neoplasms↗

PubMed1999

Induced production of nitric oxide and sensitivity of alveolar macrophages derived from mice with different sensitivity to Coxiella burnetii.

We compared in vitro sensitivities to Coxiella burnetii of alveolar macrophages, derived from mice sensitive and resistant to C. burnetii, respectively, and examined the role of nitric oxide (NO) in the C. burnetii infection. Alveolar macrophages of sensitive A/J mice showed a larger population of C. burnetii antigen-positive cells than those of resistant C57BL/6 mice. C. burnetii induced NO production in alveolar macrophages, but N-methyl-L-arginine and sodium nitroprusside (SNP), NO inhibitor and donor, respectively, did not inhibit the infection. Thus the NO induction seems to be independent of the cell defense mechanism against the C. burnetii infection.

Animals↗

PubMed1999

Expression and possible functional role of the beta3-adrenoceptor in human and rat detrusor muscle.

PURPOSE: To investigate the presence of the beta3-adrenoceptor (beta3-AR) in human and rat detrusor muscle and the usefulness of beta3-AR agonists as drugs for the treatment of urinary frequency. MATERIALS AND METHODS: FK175, ethyl [(S)-8-[(R)-2-(3-chlorophenyl)-2-hydroxyethylamino]-6,7,8,9-tetrah ydro-5H-benzocyclohepton-2-yloxy]acetate monohydrochloride monohydrate, was used as a beta3-AR selective agonist. The expression of beta-AR subtypes (beta1-, beta2-, beta3-AR) mRNA was investigated in rat and human detrusor muscle by RT-PCR. Beta3-AR agonist induced cyclic AMP (cAMP) levels were measured in rat detrusor muscle strips. The relaxation response produced by a beta3-AR agonist was measured in a KCl induced tonic contraction model in rat detrusor muscle strips. The effect of a beta3-AR agonist on urinary bladder function was investigated by cystometry using a conscious rat model of urinary frequency. RESULTS: beta3-AR mRNA was substantially expressed in both rat and human detrusor muscles. The beta3-AR agonist, FK175 (10(-7) M), increased the cAMP level by 30% in rat detrusor muscle. In isolated rat detrusor muscle strips contracted with KCl, the beta3-AR agonist, FK175 (10(-8) to 10(-4) M), produced a concentration-dependent relaxation. Moreover, although the relaxation induced with FK175 was blocked by the non-selective beta-AR antagonist, bupranolol, it was unaffected by ether the beta1-AR selective antagonist, CGP 20712A, or the beta2-AR selective antagonist, ICI 118551, suggesting that FK175 induced the relaxation via the beta3-AR. Furthermore, in the rat model, the orally administered beta3-AR agonist, FK175 (10 mg./kg.) significantly increased bladder capacity with no change of micturition pressure or threshold pressure. CONCLUSIONS: These results suggest that beta3-AR agonists may be effective in the treatment of urinary frequency.

Adrenergic beta-Agonists↗

PubMed1998

Identification of multidrug resistant protein 1 of mouse leukemia P388 cells on a PVDF membrane using 6-aminoquinolyl-carbamyl (AQC)-amino acid analysis and World Wide Web (WWW)-accessible tools.

Multidrug resistant protein 1 (MDR1) in a doxorubicin-resistant mouse leukemia cell line (P388/DOX) was identified using its amino acid composition combined with protein database searching (ExPASy and EMBL PROPSEARCH) via the World Wide Web. The proteins were separated by one-dimensional SDS-polyacrylamide gel electrophoresis, blotted onto a polyvinylidene fluoride membrane, and stained with Coomassie brilliant blue. A 160-kDa protein band was acid-hydrolyzed in the vapor phase (6 N HC1) and converted to 6-aminoquinolyl-carbamyl (AQC)-amino acids without extraction of the amino acids from the membrane. The amino acid composition of the protein was determined using the sensitive AQC-amino acid analysis method, improving our previously described method. The improved method involved using a Cosmosil 5C8-MS column instead of a Pegasil C8; replacement of the mobile phase A, constituent, 75 mM ammonium phosphate (pH 7.5), with 30 mM sodium phosphate buffer (pH 7.2); and slight modification of the separation program (9). All manipulations for protein hydrolysis and AQC derivatization were carried out in a hood using clean tools. This minimized contamination of amino acids at the low femtomolar level. A database search was carried out with bovine serum albumin as a calibration protein. MDR1 in P388/DOX was ranked first by both databases with high reliability (score 14 for ExPASy, distance 1.34 for EMBL).

ATP Binding Cassette Transporter, Subfamily B, Mem↗

PubMed1998

Yeast positive-stranded virus-like RNA replicons. 20 S and 23 S RNA terminal nucleotide sequences and 3' end secondary structures resemble those of RNA coliphages.

Saccharomyces cerevisiae strains carry single-stranded RNAs called 20 S RNA and 23 S RNA. These RNAs and their double-stranded counterparts, W and T dsRNAs, have been cloned and sequenced. A few nucleotides at both ends, however, remained unknown. These RNAs do not encode coat proteins but their own RNA-dependent RNA polymerases that share a high degree of conservation to each other. The polymerases are also similar to the replicases of RNA coliphages, such as Qbeta. Here we have determined the nucleotide sequences of W and T dsRNAs at both ends using reverse transcriptase polymerase chain reaction-generated cDNA clones. We confirmed the terminal sequences by primer-extension and RNase protection experiments. Furthermore, these analyses demonstrated that W and T dsRNAs and their single-stranded RNA counterparts (i) are linear molecules, (ii) have identical nucleotide sequences at their ends, and (iii) have no poly(A) tails at their 3' ends. Both 20 S and 23 S RNAs have GGGGC at the 5' ends and the complementary 5-nucleotides sequence, GCCCC-OH, at their 3' ends. S1 and V1 secondary structure-mapping of the 3' ends of 20 S and 23 S RNAs shows the presence of a stem-loop structure that partially overlaps with the conserved 3' end sequence. Nucleotide sequences and stem-loop structures similar to those described here have been found at the 3' ends of RNA coliphages. These data, together with the similarity of the RNA-dependent RNA polymerases encoded among these RNAs and RNA coliphages, suggest that 20 S and 23 S RNAs are plus-strand single-stranded virus-like RNA replicons in yeast.

Cloning, Molecular↗

PubMed1998

Inactivation of human manganese-superoxide dismutase by peroxynitrite is caused by exclusive nitration of tyrosine 34 to 3-nitrotyrosine.

Peroxynitrite has recently been implicated in the inactivation of many enzymes. However, little has been reported on the structural basis of the inactivation reaction. This study proposes that nitration of a specific tyrosine residue is responsible for inactivation of recombinant human mitochondrial manganese-superoxide dismutase (Mn-SOD) by peroxynitrite. Mass spectroscopic analysis of the peroxynitrite-inactivated Mn-SOD showed an increased molecular mass because of a single nitro group substituted onto a tyrosine residue. Single peptides that had different elution positions between samples from the native and peroxynitrite-inactivated Mn-SOD on reverse-phase high performance liquid chromatography were isolated after successive digestion of the samples by staphylococcal serine protease and lysylendopeptidase and subjected to amino acid sequence and molecular mass analyses. We found that tyrosine 34 of the enzyme was exclusively nitrated to 3-nitrotyrosine by peroxynitrite. This residue is located near manganese and in a substrate O-2 gateway in Mn-SOD.

Amino Acid Sequence↗

PubMed1998

The relation between the growth patterns of gastric carcinoma and the expression of hepatocyte growth factor receptor (c-met), autocrine motility factor receptor, and urokinase-type plasminogen activator receptor.

BACKGROUND: Hepatocyte growth factor receptor (c-met), autocrine motility factor receptor (AMFR), and urokinase-type plasminogen activator receptor (uPAR) are known to play important roles in tumor cell migration, invasion, and metastasis. The authors studied the relation between the expression patterns of these genes and the growth patterns of human gastric carcinoma. METHODS: The relation between the expression of c-met, AMFR, and uPAR and clinicopathologic parameters was studied using immunohistochemical preparations from 102 paraffin embedded primary gastric carcinomas. RESULTS: Of 102 cases, 43 (42%) had overexpression of c-met, and AMFR and uPAR immunoreactivity was observed in 41 cases (40%) and 38 cases (37%), respectively. Macroscopic examination revealed that all three genes were expressed in 1 (3%) of 32 early stage gastric carcinomas, 0 (0%) of 29 localized carcinomas (Borrmann types 1 and 2), and 16 (39%) of 41 infiltrating carcinomas (Borrmann types 3 and 4). In particular, the incidence (68%, 13 of 19 cases) of simultaneous expression of the three genes was significantly higher in Borrmann type 4 gastric carcinoma than in the other macroscopic types (P < 0.01). The overexpression of these genes was also closely associated with lymph node metastasis and peritoneal dissemination. In addition, the simultaneous overexpression of the three genes was associated with positive lymphatic vessel invasion and infiltrating type. Patients with tumors that simultaneously expressed all three genes had significantly poorer prognoses than those with tumors expressing only one or two of the genes. Furthermore, the number of genes expressed was closely related to the prognosis, and the Cox proportional hazards model identified this as one of the independent prognostic factors. CONCLUSIONS: These results suggest that the expression patterns of c-met, AMFR, and uPAR may be closely associated with the progression and invasion of gastric carcinoma as well as the prognoses of the patients. Borrmann type 4 gastric carcinoma is characterized by the diverse and simultaneous expression of these three genes.

Adult↗