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Biomedical subjects

N Ohno

Publications and source records attributed to N Ohno.

At least 73 records · Page 4Linked to original sources
PubMed1999

Gene therapy by transforming growth factor-beta receptor-IgG Fc chimera suppressed extracellular matrix accumulation in experimental glomerulonephritis.

BACKGROUND: The evidence that transforming growth factor-beta (TGF-beta) is a key mediator in the pathogenesis of fibrotic diseases is now supported by several lines of investigation. This evidence provides a certain base for targeting TGF-beta as an antifibrotic agent. METHODS: We generated a chimeric cDNA, termed TGF beta RII/Fc, encoding an extracellular domain of the TGF-beta type II receptor fused to the IgG-Fc domain, and tested whether TGF beta RII/Fc could be a novel strategy for treating glomerular diseases. RESULTS: In cultured BNul-7 cells, recombinant TGF beta RII/Fc reversed the antiproliferative response induced by TGF-beta 1. In addition, TGF beta RII/Fc diminished the TGF-beta 1-induced production of EIIIA-positive fibronectin in cultured normal rat kidney cells. We then introduced the chimeric cDNA into the muscle of the nephritic rats by the hemagglutinating virus of Japan liposome-mediated gene transfer method in order to block the TGF-beta activity in nephritic glomeruli through systemic delivery of chimeric molecules. Treatment with TGF beta RII/Fc gene transfection could suppress the glomerular TGF-beta mRNA in nephritic rats with a comparable effect in the reduction of extracellular matrix accumulation. CONCLUSION: TGF beta RII/Fc successfully inhibited the action of TGF-beta in vitro and in vivo, and gene therapy by chimeric TGF beta RII/Fc might be feasible for the therapy of glomerulosclerosis.

Animals↗
PubMed1999

Electron microscopic observation on the parathyroid gland of the cardiomyopathic hamster (UM-X7.1 Syrian hamster).

The ultrastructure of the parathyroid gland (PTG) of the UM-X7.1 cardiomyopathic hamster (the UM-X7.1) was qualitatively and quantitatively compared with that of the normal hamster. In the UM-X7.1, the Golgi apparatus and rER were well developed compared with those of the control hamster. Morphometric analysis revealed that the mean values of the Golgi apparatus, rER and large vacuolar bodies were significantly greater in the UM-X7.1 than in the normal hamster and the mean value of lipid droplets was significantly less in the UM-X7.1 than in the normal hamster. Ultrastructurally, we consider that in the UM-X7.1, the synthesis and release of the parathyroid hormone may be activated by an excessive amount of circulating catecholamine, and the functional activity of the PTG may be higher than that in the normal hamster.

Animals↗
PubMed1999

Highly specific detection of H2O2-dependent luminol chemiluminescence in stimulated human leukocytes using polyvinyl films.

When human polymorphonuclear leukocytes (PMN) were attached to glass coverslips, cells always spread and formed reactive oxygen species prior to any experimental stimulation. To avoid this, a polyvinylidine chloride film was used as an inactive substance to place the cells. Cells engaged in phagocytosis on the film exhibited a specific H2O2-mediated luminol chemiluminescence (LCL) at the cell-particle interface; the cells stimulated with 12-O-tetradecanoylphorbol-13-acetate became aggregated and the LCL was observed at the cell-cell contact. These results corresponded well with those obtained by an electron microscopic H2O2-demonstration method.

Humans↗
PubMed1999

Antigen-specific response of murine immune system toward a yeast beta-glucan preparation, zymosan.

Zymosan, a particulate beta-glucan preparation from Saccharomyces cerevisiae, shows various biological activities, including anti-tumor activity. We have previously shown that soluble beta-glucan initiated anti-tumor activity was long-lived and was effective even by prophylactic treatment at 1 month prior to tumor challenge. However, the activity by zymosan was relatively short-lived. Antigen-specific responses of mice to zymosan might be a causative mechanism. In this paper, mice were immunized with zymosan and antibody production and antigen-specific responses of lymphocytes to zymosan were analyzed. Sera of zymosan immune mice contained zymosan-specific IgG assessed by enzyme-linked immunosorbent assay and FACS. Spleen and bone marrow cells of zymosan-immune mice showed higher cytokine production in response to zymosan. Specificity of zymosan-specific responses were also analyzed using various derivatives prepared from zymosan. These facts strongly suggested that mice recognize zymosan as antigen in addition to non-specific immune stimulant.

Animals↗
PubMed1999

Increase of hematopoietic responses by triple or single helical conformer of an antitumor (1-->3)-beta-D-glucan preparation, Sonifilan, in cyclophosphamide-induced leukopenic mice.

It has been suggested that the immunopharmacological activity of soluble (1-->3)-beta-D-glucan depends on its conformation in mice. In this study, we examined the relationship between the conformation of Sonifilan (SPG) and hematopietic responses in cyclophosphamide (Cy)-induced leukopenic mice. SPG, a high molecular weight (1-->3)-beta-D-glucan, has a triple helical conformation in water, and it was changed by treatment with aqueous sodium hydroxide to the single helical conformer (SPG-OH). The effects of SPG or SPG-OH on hematopoietic responses in cyclophosphamide induced leukopenic mice were investigated by monitoring i) gene expression of cytokines by RT-PCR, ii) protein synthesis of interleukin 6 (IL-6) by ELISA and iii) colony formation of bone marrow cells (BMC). The mice administered Cy and SPG or SPG-OH expressed and produced higher levels of IL-6 mRNA and protein than the mice administered only Cy. Gene expression of NK1.1 was also induced by Cy/SPG (or SPG-OH) treatment. Induced gene expression of stem cell factor (SCF) and macrophage-colony stimulating factor (M-CSF) by SPG/SPG-OH were also found in in vitro culture of BMC from Cy treated mice. These results strongly suggested that conformation of the glucans, single and triple helix, are independent of the hematopietic response.

Animals↗
PubMed1999

Inhibition of tumor cell proliferation by type IV collagen requires increased levels of cAMP.

Previous studies from our laboratories demonstrated that a peptide from the noncollagenous domain of the alpha3 chain of basement membrane collagen (COL IV), comprising residues 185-203, inhibits polymorphonuclear leukocyte activation and melanoma cell proliferation; this property requires the presence of the triplet -SNS- in residues 189-191 (Monboisse et al., J. Biol. Chem., 269, 25475, 1994; Han et al., J. Biol. Chem., 272, 20395, 1997). In the present study, we demonstrate that whole native COL IV and -SNS- containing synthetic peptides (10 microg/ml) added to culture medium inhibit the proliferation of not only melanoma cells, but also breast-, pancreas- and stomach-tumor cells up to 67%, and prostate tumor cells by 15%. ALC-COL IV at 5 microg/ml was shown to inhibit melanoma cell proliferation maximally at 69% and the alpha3(IV)185-203 peptide inhibited proliferation (62%) maximally at 10 microg/ml. Treatment of the alpha3(IV)185-203 peptide with either a specific mAb or a polyclonal antibody, prepared against the sequence alpha3(IV)179-208, decreased the ability of the peptide to inhibit cell proliferation by 97%, while treatment of ALC-COL IV with the same antibodies inhibited proliferation by 44%. Exposure of the above tumor cells to COL IV or the peptides resulted in an increase of intracellular cAMP that was inhibited by prior treatment of the protein with the above antibodies. To investigate the role of cAMP in the inhibition of cell proliferation, cAMP analogs and inhibitors were used. cAMP analogs mimicked the inhibitory effect of the peptide. Rp-cAMPS, a cAMP competitive inhibitor, suppressed the inhibitory effect of ALC-COL IV and of the cAMP analogs. The protein kinase-A inhibitor H-89 blocked the ability of ALC-COL IV and of the alpha3(IV)185-203 peptide to inhibit tumor cell proliferation. These data suggest that ALC-COL IV, through its alpha3(IV) chain, inhibits tumor cell proliferation utilizing a signal transduction pathway which includes cAMP and cAMP-dependent protein kinase(s).

Amino Acid Sequence↗
PubMed1998

Contribution of CR3 to nitric oxide production from macrophages stimulated with high-dose of LPS.

The contribution of the complement receptor type 3 (CR3) to nitric oxide (NO) production from macrophages stimulated by LPS was investigated. When thioglycollate-elicited mouse peritoneal macrophages were stimulated with a high dose of LPS (10 micrograms/ml) in both the presence and absence of fetal calf serum, a source of LPS binding protein (LBP) necessary for the binding of LPS to CD14, NO production was observed. These findings suggest that CD14-dependent and CD14-independent signaling pathways for NO production are present in macrophages. Because binding and phagocytosis of bacteria by macrophages through the CR3 has been previously reported, we investigated whether the CR3 acts in CD14-independent signaling pathway for NO production. By flow cytometric analysis, the binding of FITC-labeled anti-CR3 monoclonal antibody (anti-CR3 mAb) to macrophages was inhibited by LPS. Anti-CR3 mAb induced iNOS protein and produced NO in a dose dependent manner. Further, NO production induced by anti-CR3 mAb was also inhibited by zymocel, beta-glucan with a high affinity to CR3. These results suggest that the CR3 molecule acts in a CD14-independent signaling pathway, and contributes to NO production by macrophages stimulated with high doses of LPS.

Animals↗
PubMed1998

Mother-to-infant transmission occurs more frequently with GB virus C than hepatitis C virus.

A total of 107 hepatitis C virus (HCV)-infected pregnant women were screened for GB virus C (GBV-C) RNA in their sera, and 11 (10.3%) were positive. Among 11 infants born to these HCV/GBV-C co-infected mothers, GBV-C RNA was detected in 7 (63.6%) while HCV RNA was found in 1 (9.1%) within 1 year after birth: this difference was statistically significant (p = 0.023). The mothers of infected infants had significantly higher serum titers of GBV-C RNA than those of uninfected infants: 10(6.7 +/- 0.5) vs 10(4.0 +/- 1.0) copies/ml in average (p = 0.001). The baby in whom HCV RNA was found was also positive for GBV-C RNA, and had an elevation in serum transaminase levels, whereas all the other GBV-C infected infants showed no evidence for hepatitis. A family study, performed on 2 of the 7 infected cases, revealed that all the elder siblings of the index infants were also GBV-C RNA-positive. Nucleotide sequence of GBV-C RNA, amplified by PCR from an NS3 region, was completely identical between the mother and the infant within each family, but varied significantly across different families. These results suggest that GBV-C is more easily transmitted from mother to infant than HCV, although hepatitis is not caused thereby.

Base Sequence↗
PubMed1998

Autoblood plus OK432 pleurodesis with open drainage for persistent air leak after lobectomy.

Prolonged air leak with dead space after lobectomy was successfully treated with autoblood plus OK432 pleurodesis in 10 patients. We tried this method for the purpose of (1) reduction of dead space and (2) effective chemical sclerosis. This easy method was associated with acceptable side effects and should be considered as an option for the treatment of prolonged air leak after lobectomy, especially with dead space.

Blood↗
PubMed1998

Fluoride profiles of perikymata in enamel surfaces of human premolars.

Twenty-five premolars (from Nagoya, Japan < 0.1 parts/10(-6)F in drinking water) were sampled to determine the fluoride content in imbrication lines of Retzius between the grooves and ridges of perikymata on the enamel surface. Eight small windows were formed on each surface in groove and ridge regions using an etched microsampling technique. By using a regression curve, y = ax(-b), fluoride concentrations were compared at depths of 1, 3, 5, 10, 20, 30 and 50 microm in the perikymata regions. Fluoride concentrations increased gradually from the age of 10 to 12 years on the premolar surfaces, but were significantly higher in 12-year-olds or older (erupted teeth) than in 10-year-olds or younger (unerupted teeth). No obvious difference in fluoride concentrations was found between males and females. In the outermost enamel ( < 5 microm depth), fluoride concentrations were significantly higher in the grooves than the ridges of the perikymata. Comparison of the exponential regression coefficients (-b) of the fluoride profiles showed a significant difference between the grooves and ridges. It was concluded that fluoride concentrations and profiles were higher in grooves than in ridges of perikymata, probably because they are naturally porous and are stagnation areas attracting dental plaque.

Acid Etching, Dental↗
PubMed1998

Role of cadherin internalization in hydrogen peroxide-mediated endothelial permeability.

Exposure of endothelial monolayers to hydrogen peroxide results in increased solute permeability in a time- and dose-dependent fashion. This effect is prevented by either staurosporine, an inhibitor of PKC, or by Gö6976, an inhibitor of "classical" PKC isoforms. Immunohistochemistry of peroxide-treated monolayers illustrates a loss of cadherin staining at cell junctions and gap formation predominantly at tri-cellular junctions. Both staurosporine and Gö6976 prevented peroxide-induced gap formation. Peroxide also stimulated internalization of cadherins as measured by the trypsin protection assay, which was not blocked by staurosporine or Gö6976. These data suggest that peroxide causes: 1) a time- and dose-dependent increase in permeability and dose-dependent increase in gap formation, both of which are PKC dependent; and 2) promotes PKC-independent cadherin internalization. These data indicate that cadherin internalization may be part of the mechanism through which oxidants regulate solute permeability.

Animals↗
PubMed1998

Cytokine synthesis of human monocytes stimulated by triple or single helical conformer of an antitumour (1-->3)-beta-D-glucan preparation, sonifilan.

It has been suggested that the immunopharmacological activity of soluble (1-->3)-beta-D-glucan depends on it's conformation in mice. In this study, we examined the relationship between the conformation of Schizophyllan (SPG), a high molecular weight (1-->3)-beta-D-glucan, and cytokine productivity in an in vitro human system. Monocyte-like human cell lines, THP-1 and U-937, and peripheral blood mononuclear cells (PBMC) were used. THP-1 and U-937 cells were differentiated by phorbol myristate acetate (PMA) before use. SPG usually has a triple helical conformation in water, but it was modified by treatment with aqueous sodium hydroxide to become a single helical conformer (SPG-OH). SPG or SPG-OH was added to the macrophage cell culture and gene expression and translation of several cytokines was analyzed by RT-PCR, ELISA, or bioassays. Differentiated THP-1 expressed high levels of cytokine genes, such as IL-8, in response to SPG-OH. High levels of IL-12 p70 were detected from THP-1 cells stimulated with SPG-OH. U-937 cells expressed high levels of IL-8 and TNF-alpha after SPG-OH treatment. Furthermore, PBMC isolated from healthy donors also strongly reacted with SPG-OH but not with SPG. High concentrations of TNF-alpha were detected in SPG-OH-stimulated PBMC cultures. These data suggest that the biological activities of SPG are strongly associated with its conformation in humans.

Adjuvants, Immunologic↗
PubMed1998

Effect of O-antigenic polysaccharide of Escherichia coli on endotoxin neutralizing activity of lysozyme.

Endotoxemia is considered to be associated with the high mortality of gram-negative septic patients. Increasing evidence shows that beta-lactam antibiotics have a propensity to induce endotoxin release from the bacterial outer membrane while killing bacteria. We have recently found that egg white lysozyme (EW-LZM) shows strong inhibition of beta-lactam induced bacteriolysis and lipopolysaccharide (LPS) release from Escherichia coli O111, resulting in reduction of the LPS-initiated inflammatory response. In this study, we compared the effect of EW-LZM on E. coli J5, which possesses rough-type LPS (RaLPS), in order to demonstrate the effect of O-antigenic polysaccharide on endotoxin neutralizing activity of EW-LZM and on inhibition of beta-lactam induced lysis by LZM. Both of the beta-lactam induced bacterial lysis and subsequent LPS release were almost completely inhibited by EW-LZM. The effect was more potent than that of wild-type LPS as assessed by released LPS concentration and LPS induced cytokine syntheses. In addition, EW-LZM was effective against lethal infection of E. coli J5 in cyclophosphamide induced leukopenic mice. These facts strongly suggested that O-antigenic polysaccharide negatively modulates LPS neutralizing activity of EW-LZM.

Animals↗
PubMed1998

Gradual solubilization of Candida cell wall beta-glucan by oxidative degradation in mice.

Candida spp. is a medically important fungi which induces disseminated candidiasis and candidemia in hospitalized immunocompromised patients. The cell wall of Candida is mainly composed of two polysaccharides, mannan and beta-glucan, and at least part of beta-glucan is basically insoluble in H2O or NaOH. We became interested in when and how particulate beta-glucan changes to the soluble form. However, the fate of wall components has not been examined in detail. In this study, modification and solubilization of the cell wall beta-glucan were analyzed in vivo and in vitro. Cells of Candida, intravenously administered to mice (1 mg/mouse), were immediately deposited mainly in liver as determined by 3H-labeled cells. Beta-Glucans were detected in these mice for at least for 6 months by the beta-glucan specific assay. During this period, the insoluble cell wall beta-glucan was gradually solubilized in these organs, probably by oxidative stress of macrophages. Candida cells and particulate beta-glucans were also gradually solubilized in vitro using sodium hypochlorite solution, but part of the cell wall beta-glucan was still insoluble even after treatment with concentrated hypochlorite solution for one day at room temperature. These findings strongly suggested that the fungal cell wall beta-glucans were quite resistant to oxidative metabolism in vivo and in vitro, and thus deposited for quite long period in the host.

Animals↗
PubMed1998

Immunotoxicity of soluble beta-glucans induced by indomethacin treatment.

(1 --> 3)-Beta-D-Glucan (beta-glucan) is a biological response modifier that regulates host immune response. However, the side effects of this drug have not been extensively examined. In this study, we found that the combination of a beta-glucan and a nonsteroidal anti-inflammatory drug, indomethacin, induced lethal toxicity in mice. Lethal toxicity of orally administered indomethacin (multiple administration to ICR mice; once a day for 2 weeks) was 0/8 (2.5 mg kg(-1)) and 5/8 (5 mg kg(-1)) (death/total) over 2 weeks. The toxicity was enhanced to 3/8 and 8/8 in mice treated with a clinical beta-glucan preparation, sonifilan (250 microg/mouse, single i.p. administration on day 0). A similar effect was observed for other beta-glucans, including SSG, grifolan, zymosan A and zymocel. Enhanced lethal toxicity resulted from a single p.o. administration of indomethacin on day 5 to day 9 after multiple beta-glucans administration. Interferon-gamma, interleukin-6 and colony stimulating factor concentrations in sera of indomethacin/beta-glucan-treated mice were significantly elevated. These results strongly suggest that indomethacin/beta-glucan treatment induces lethality in mice by maladjusting the cytokine network.

Adjuvants, Immunologic↗
PubMed1998

Soluble mannan and beta-glucan inhibit the uptake of Malassezia furfur by human monocytic cell line, THP-1.

The uptake of live and heat-killed Malassezia furfur HIC 3321, HIC 3343 and Candida albicans ATCC 10231 by human monocytic cell line, THP-1, was examined. THP-1 was differentiated by PMA for 7 days before use. The uptake of these yeasts by THP-1 was increased in a concentration-dependent manner of yeasts, and the uptake reached plateau level at the E/T (yeast/THP-1) ratio 5. In addition, a higher percentage of heat-killed cells than live cells was taken in THP-1. Yeast mannan and beta-1,3-glucan, random coiled conformer, inhibited the uptake of live and heat-killed M. furfur by THP-1, though dextran T-250, that is alpha-glucan, and schizophyllan (SPG), triple helix conformer of beta-glucan, did not. Interestingly, mannan inhibited the uptake of both types, live and heat-killed, of C. albicans, however, laminaran inhibited the uptake of heat-killed C. albicans alone. Opsonization of these yeasts with normal human serum enhanced the uptake of yeasts, although opsonization with heat-inactivated serum, the treatment at 56 degrees C for 30 min, did not enhance. These results suggested that live and heat-killed M. furfur was recognized by THP-1 through mannose receptor, beta-glucan receptor and complement receptor type 3 via the activation of alternative pathway of complement.

Candida albicans↗