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T Oikawa

Publications and source records attributed to T Oikawa.

At least 91 records · Page 5Linked to original sources
PubMed1999

Stereoisomers of glutathione: preparation and enzymatic reactivities.

We synthesized a series of stereoisomers of glutathione (GSH) and glutathione disulfide (GSSG) by the solid-phase method. These peptides were used to examine their reactivities with enzymes acting on glutathione. The glutathione reductase of yeast acted only on LL-GSSG. Glutathione S-transferase catalyzed the conjugation of 1-chloro-2,4-dinitrobenzene with LL-GSH and DL-GSH (Km (mM): for LL-GSH, 0.035; and for DL-GSH, 0.62), but the DD- and LD-diastereomers were inert. gamma-Glutamyl transpeptidase catalyzed the transfer of gamma-glutamyl moiety of LL-GSH and DL-GSH to taurine forming gamma-glutamyl taurine and cysteinyl taurine (Km (mM): for LL-GSH, 0.336; and for DL-GSH, 0.628), but the other diastereomers were not the substrates. The occurrence of L-cysteinyl residue in the tripeptides is required for the glutathione analogue to be a substrate of the enzymes.

Chromatography, High Pressure Liquid↗
PubMed1999

[Electroconductive lithotripsy: clinical results of the Sonolith sigma].

The electroconductive lithotripter (ECL) is a new concept for shockwave generation in which a highly conductive solution channels the discharge between the anode and cathode. Out of 152 patients treated, complete follow up data were available on 151 patients. The average number of shocks per treatment was 2,138. At 3 months the overall stone-free rate was 73.5%. Success rate, defined as stone-free or asymptomatic residual fragments measuring 4 mm or less, was 84.9% for renal and 94.9% for ureteral calculi. The overall success rate for all calculi was 89%.

Adult↗
PubMed1999

Electron microscopic peritubular capillary lesions: a new criterion for chronic rejection.

Typical chronic rejection showing prominent glomerular and/or arterial lesions is less common in renal allograft patients treated with cyclosporin A (CsA). We investigated the value of peritubular capillary lesions as a criterion for chronic rejection in the CsA era. A total of 129 renal graft biopsies, taken from recipients showing graft dysfunction after more than 2 months post-operatively, were examined by electron microscopy, and peritubular capillary lesions were studied, especially multi-layered basement membrane lesions (MLPTC). Eighty-two biopsy specimens taken from non-transplantation patients were also studied as a control. Five biopsies (6%) showed mild and atypical MLPTC in the control group. Of the 129 allograft biopsies, MLPTC was seen in 55 (42.6%). The prevalence showed no significant relationship to the interval from operation to biopsy. MLPTC was either typical (n = 10) or incomplete (n = 45). Concomitant membrane disruption, edema and lymphocyte infiltration of the subendothelial space, reflecting acute cellular rejection, were occasionally noted in both groups. Incomplete MLPTC often developed within 1 yr after surgery, in association with acute rejection. It was found in 26-50% of biopsies at any time up to 5 yr post-operatively. The incidence of typical MLPTC was 5.7-12.8% over 1 yr post-operatively. These findings suggest that the development of chronic rejection is closely related to relapsing acute tubulo-interstitial allograft rejection which is often clinically silent. We concluded that MLPTC is useful as a specific criterion for chronic rejection.

Basement Membrane↗
PubMed1999

A case of IgA nephropathy after ABO-incompatible living kidney transplantation.

A 39-yr-old Japanese man underwent living related kidney transplantation. Because the graft was ABO-incompatible, he was treated with double filtration plasmapheresis before transplantation and his immunosuppressive therapy was stronger than usual. However, immunoglobulin A nephropathy, accompanied by cellular crescents and necrotizing lesions, developed after 18 months. To our knowledge, the association of IgA nephropathy with ABO-incompatible kidney transplantation has not been reported previously.

ABO Blood-Group System↗
PubMed1998

Reduction of DNA binding activity of the GATA-1 transcription factor in the apoptotic process induced by overexpression of PU.1 in murine erythroleukemia cells.

Previously we have shown that overexpression of PU.1, an Ets family transcription factor, in murine erythroleukemia (MEL) cells results in apoptotic cell death in the presence of the differentiation-inducing reagent dimethyl sulfoxide (DMSO). In this study, we examined the dynamics of GATA-1 and NF-E2 hematopoietic transcription factors during the induction of apoptosis, because GATA-1 has been shown to be implicated in survival of erythroid cells. Formation of the GATA-1-DNA complex as judged by EMSA was markedly reduced when apoptosis was induced, although subcellular localization of the GATA-1 protein and expression levels of the GATA-1 mRNA and protein were not changed during the apoptotic process. Complex formation was not reduced when apoptosis was avoided by adding 30% serum in culture medium and when mutant PU.1 proteins with the deletion of the DNA-binding (Ets) or transactivation domain were expressed. Complex formation in nuclear extracts of parental MEL cells was reduced when they were mixed with those of apoptotic cells, suggesting that apoptotic cells may contain a factor(s) preventing GATA-1 from binding to DNA. In contrast to GATA-1, formation of the NF-E2-DNA complex was not changed during the process of apoptosis, although the expression level of the NF-E2 p45 gene was reduced in the process. These results suggest that reduction of the DNA-binding activity of GATA-1 may partly account for PU.1-mediated apoptosis in MEL cells.

Animals↗
PubMed1998

Down-regulation of c-myc and bcl-2 gene expression in PU.1-induced apoptosis in murine erythroleukemia cells.

We found that over-expression of PU.1, a member of the ets family of transcription factors, induces apoptotic cell death along with differentiation of DMSO stimulation in murine erythroleukemia (MEL) cells. To elucidate the molecular mechanisms of apoptosis, cell-cycle distribution and expression of several genes encoding apoptosis-promoting and -inhibiting factors were analyzed during the process of PU.1-induced apoptosis. FACS analysis revealed that cells were accumulated in the G0/G1 phase of the cell cycle before apoptosis. Morphological analysis of PI-stained nuclei of the apoptotic cells sorted by a FACScan showed 22.6% in G0/G1, 35.8% in S and 8.5% in G2/M phase by fluorescent microscopy after cell sorting, suggesting that PU.1-induced apoptosis in MEL cells occurs in G0/G1 through S phases. Semi-quantitative RT-PCR revealed that expression of c-myc and bcl-2 genes was reduced during the apoptotic process, while expression of bax and bcl-X(L) genes was not changed. Expression of the p53 gene was reduced rather than enhanced, suggesting that PU.1-induced apoptosis in MEL cells is p53-independent. Apoptosis was inhibited by adding 30% serum in culture, while no reduction of c-myc and bcl-2 gene expression was observed. Forced expression of the c-myc, bcl-2 and bcl-X(L) genes protected MEL cells from apoptosis. Our results suggest that a reduction of at least 2 important apoptosis-inhibiting factors, c-Myc and Bcl-2, is involved in PU.1-induced apoptosis in MEL cells.

Animals↗
PubMed1998

The proteasome is involved in angiogenesis.

The proteasome is a recently identified intracellular protease whose catalytic active site is a threonine residue and has been shown to play key roles in a variety of important intracellular events, including cell cycle progression, the antigen-presenting pathway, and apoptosis. However, its biological significance in multicellular organisms is still largely unknown because of lack of experimental systems for its study. Here we verified potential involvement of the proteasome in angiogenesis using lactacystin, a specific proteasome inhibitor. Lactacystin treatment resulted in almost complete prevention of in vivo neovascularization in the developing chick embryo chorioallantoic membrane. It also inhibited vascular endothelial tube formation on Matrigel, a model for in vitro angiogenesis, in a concentration-dependent fashion. Moreover, it prevented production of plasminogen activator, an important protease responsible for induction of angiogenesis, by endothelial cells, which correlated well with its suppression of intracellular proteasome activity. Our studies suggest that the proteasome operates in the process of angiogenesis, a phenomenon essential in important physiological and pathological settings.

Acetylcysteine↗
PubMed1998

Fine structure of three types of olfactory organs in Xenopus laevis.

There is no report on the fine structure of three types of olfactory organs in Xenopus laevis. Their functional assignments in olfaction are not yet established. The fine structure of three types of olfactory organs, olfactory epithelium (OE), vomeronasal organ (VNO), and middle chamber epithelium (MCE), was examined in Xenopus laevis by light and electron microscopy. The olfactory cells of the OE and the sensory cells of the VNO were equipped with cilia and microvilli, respectively, similar to terrestrial animals that possess both the OE and the VNO. On the other hand, the sensory cells of the MCE were classified into two types, the sensory cells with cilia and the sensory cells with microvilli, like those of the OE in fish. These findings suggest that the OE and the VNO in Xenopus laevis detect different kinds of odoriferous molecules in air, whereas the MCE is involved in the perception of odorants in water.

Animals↗
PubMed1998

Enhanced expression of the urokinase-type plasminogen activator gene and reduced colony formation in soft agar by ectopic expression of PU.1 in HT1080 human fibrosarcoma cells.

To investigate the cell biological function of PU.1, a member of the Ets family of transcription factors, a vector capable of expressing the protein was transfected into HT1080 human fibrosarcoma cells. Exogenous expression of PU.1 in HT1080 cells reduced colony-forming efficiency but stimulated cell migration in soft agar, although it did not affect cell growth in adherent culture. Expression of the urokinase-type plasminogen activator (uPA) mRNA, which is known to be correlated with cell migration and invasion, was enhanced in PU.1 transfectants compared with mock transfectants. Run-on analysis demonstrated that uPA transcription was unaffected by PU.1, suggesting that this enhancement mainly occurs at a post-transcriptional level. On the other hand, treatment of HT1080 cells with the synthetic glucocorticoid dexamethasone (DEX; 10(-7) M) significantly reduced uPA gene expression at a transcriptional level. Furthermore, DEX inhibited cell migration in soft agar without affecting cell growth. These negative effects of DEX on uPA expression and cell migration were alleviated by the expression of PU.1 in HT1080 cells, whereas expression of the N-ras oncogene, which is responsible for maintenance of the transformed phenotypes in HT1080 cells, was unaffected by PU.1 expression or DEX treatment in the cells. Our results suggest that expression of PU.1 can stimulate uPA gene expression at the post-transcriptional level, which may subsequently lead to activation of cell motility and/or reduced cell-cell adhesion, but reduces anchorage-independent growth of HT1080 cells.

Agar↗
PubMed1998

Endo-beta-glucanase secreted by a psychrotrophic yeast: purification and characterization.

A psychrotrophic yeast, Rhodotorula glutinis KUJ 2731, isolated from soil, effectively produced an extracellular endo-beta-glucanase (EC 3.2.1.4). The enzyme was monomeric, and the molecular mass was about 40,000 Da. The N-terminal amino acid sequence was H-Ser-Leu-Pro- Lys-Leu-Gly-Gly-Val-Asp-Leu-Ala-Gly-Leu-Asp-Ile-Gly-Lys-Asp-Lys-Asn-. alpha-Helix content was calculated to be about 32.6%. The isoelectric point was 8.57. The activation energy was 20.9 kJ/mol, which was much smaller than that of mesophilic enzymes. The enzyme was active at temperatures from 0 to 70 degrees C, with a highest initial velocity at 50 degrees C similar to other psychrotrophic enzymes. The enzyme was inhibited by Hg2+. The enzyme catalyzed hydrolysis of carboxymethyl cellulose with an apparent K(m) of 1.1% and Vmax of 556 mumol/min/mg. Products from the enzymatic hydrolysis of carboxymethyl cellulose by the enzyme were glucose, cellobiose, and cellotriose. The enzyme also catalyzed the transglycosylation of p-nitrophenyl-beta-cellotrioside to cellotetraose.

Amino Acid Sequence↗
PubMed1998

[Control of tumor-related angiogenesis].

Tumor-related angiogenes is expected to become an important target for improving treatment of cancer, because it plays key roles in tumor growth, invasion and metastasis. We considered that the successful development of such angiostatic treatment depended entirely upon the development of useful anti-angiogenic agents, and attempted to find novel angiogenesis inhibitors by using three in vivo assays, based on an idea of ours. As a result we have demonstrated that different types of agents with low molecular weight including microbial metabolites, cell differentiation modulators like retinoids and steroids, exhibit anti-angiogenic activity, anti-metastatic activity and/or antitumor activity. Taken these findings, an ideal anti-angiogenic agent is discussed.

Antineoplastic Agents↗
PubMed1998

[Trends of prostate cancer: comparison between 1986-90 and 1991-95 at Asahi General Hospital].

Patients with prostate cancer treated during 1986-95 were divided into two groups: the first five years as early period and the recent five years as late period. Background factors, therapeutic modality and prognosis were compared between these two groups. The number of patients during the late period (245) was about 1.5 fold that during the early period (161). The average age for each stage are similar within the same period, and also similar between early and late periods. Almost the same ratios were noticed on stage distribution and on histological grade between early and late periods. Therefore, the increase in the number of patients in recent years was due mainly to expansion with the same proportion of background factors. Patients with T2 and T3 during the late period received more aggressive treatments with/without endocrine therapy. Subsequently these patients seemed to show better prognosis than the patients during the early period. No differences in survival of patients with metastosis was revealed between these two periods. Therefore, the outcome was not improved by recent endocrine therapy.

Androgen Antagonists↗
PubMed1997

Overexpression of PU.1 induces growth and differentiation inhibition and apoptotic cell death in murine erythroleukemia cells.

PU.1 is a member of the ets family of transcription factors and is expressed in Friend virus-induced murine erythroleukemia (MEL) cells as a consequence of proviral integration into the PU.1/Spi-1 locus. After induction of MEL cell differentiation by treatment with dimethylsulfoxide (DMSO), expression of the PU.1/Spi-1 gene decreased before induction of beta-globin gene expression. Overexpression of PU.1 by using a zinc-inducible expression plasmid in MEL cells resulted in unexpected growth inhibition of the transfectants. When PU.1-overexpressing transfectants were treated with DMSO, growth inhibition became much pronounced and apoptosis was induced. Expression of the beta-globin gene was not induced under this condition. Neither growth inhibition nor apoptosis was induced in MEL cells after expression of mutant PU.1 proteins with a deletion of the activation domain or the DNA-binding Ets domain irrespective of the presence of DMSO. Interestingly, beta-globin gene expression was not induced in the transfectants expressing the former mutant, whereas it was induced in those expressing the latter one in the presence of DMSO. These results indicate that overexpression of PU.1 in MEL cells results in growth and differentiation inhibition and, in conjunction with DMSO treatment, apoptotic cell death. These results also suggest that the activation domain and the Ets domain of PU.1 contribute differently to induction of these effects.

Animals↗
PubMed1997

Endo-beta-Glucanase from Acetobacter xylinum: Purification andCharacterization

A cellulose-producing acetic acid bacterium,Acetobacter xylinum KU-1, abundantly produces an extracellularendo-beta-glucanase (EC 3.2.1.4) in the culture broth. The enzyme was purifiedto homogeneity by DEAE- and CM- Toyopearl 650M ion-exchange chromatography,Butyl-Toyopearl 650M hydrophobic chromatography, and Toyopearl HW-50 gelfiltration. The purified enzyme showed the maximum activity at pH 5 and50°C: it was stable up to 50°C at pH 5, activated by Co2+, andcompetitively inhibited by Hg2+; the apparentKi was 7 &mgr;M. The molecular weight of the enzyme wasdetermined to be about 39,000 by sodium dodesyl sulfate/polyacrylamide gelelectrophoresis, and about 41,000 by Toyopearl HW-50 gel filtration; theenzyme is monomeric. The enzyme hydrolyzed carboxymethylcellulose with anapparent Km of 30 mg/ml and Vmax of 1.2&mgr;M/min. It hydrolyzed cellohexaose to cellobiose, cellotriose andcellotetraose, and also cellopentaose to cellobiose and cellotriose, but didnot act on cellobiose, cellotriose, or cellotetraose.

Journal Article↗
PubMed1997

Modulation of plasminogen activator inhibitor-1 in vivo: a new mechanism for the anti-fibrotic effect of renin-angiotensin inhibition.

We examined the potential of in vivo linkage of plasminogen activator inhibitor-1 (PAI-1) and angiotensin II (Ang II) in the setting of endothelial injury and sclerosis following radiation injury in the rat. PAI-1 is a major physiological inhibitor of the plasminogen activator (PA)/plasmin system, a key regulator of fibrinolysis and extracellular matrix (ECM) turnover. PAI-1 mRNA expression in the kidney was markedly increased (9-fold) at 12 weeks after irradiation (P < 1.001 vs. normal control). In situ hybridization revealed significant association of PAI-1 expression with sites of glomerular injury (signal intensity in injured vs. intact glomeruli, P < 0.001). Angiotensin converting enzyme inhibitors (ACEI, captopril or enalapril) or angiotensin II receptor antagonist (AIIRA, L158,809) markedly reduced glomerular lesions (thrombosis, mesangiolysis, and sclerosis; sclerosis index, 0 to 4+ scale, 0.49 +/- 0.20 in untreated vs. 0.05 +/- 0.02, 0.02 +/- 0.01, 0.04 +/- 0.02 in captopril, enalapril and AIIRA, respectively, all P < 0.01 vs untreated). Further, ACEI and AIIRA markedly attenuated increased PAI-1 mRNA expression in the irradiated kidney (36, 19 and 20% expression, respectively, for captopril, enalapril and AIIRA, compared to untreated irradiated kidney, P < 0.05, < 0.01, < 0.01). This effect was selective in that neither tissue-type nor urokinase-type PA mRNA expression was affected by these interventions. Thus, we speculate that inhibition of the renin-angiotensin system may ameliorate injury following radiation by accelerating fibrinolysis and ECM degradation, at least in part, via suppression of PAI-1 expression. In summary, inhibition of Ang II, in addition to its known effects on vascular sclerosis, may also by its novel effect to inhibit PAI-1, lessen fibrosis following endothelial/thrombotic injury.

Angiotensin II↗