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Biomedical subjects

M Honma

Publications and source records attributed to M Honma.

At least 91 records · Page 5Linked to original sources
PubMed1996

Transforming growth factor beta 1 (TGF beta 1) gene expression by eosinophils in asthmatic airway inflammation.

The increase in thickness of bronchial walls by such structural changes as subepithelial fibrosis contributes to the severity and chronicity of asthma by amplifying airway narrowing. However, the pathogenesis of this structural alteration is not known. Transforming growth factor beta 1 (TGF beta 1) is known to have biologic activities relevant to the cellular and molecular events in subepithelial fibrosis, such as the deposition of collagen I and III and the increase of myofibroblasts beneath the epithelial basement membrane. Therefore, we examined TGF beta 1 gene expression in bronchial biopsy tissues from five severe asthmatics, five mild asthmatics, and five normal subjects using in situ hybridization combined with histochemical staining. Cells expressing TGF beta 1 mRNA were detected in tissues from four normal subjects, one mild asthmatic, and five severe asthmatics. The density of positive cells in severe asthmatic tissues (52.1 +/- 22.7, mean +/- SD/mm2) was significantly greater than that in mild asthmatic tissues (1.0 +/- 1.9/mm2, P < 0.01) or normal tissues (10.5 +/- 10.6/mm2, P < 0.02). The density in mild asthmatic tissues was not significantly different from that in normal tissues. The vast majority of positive cells in severe (99.1 +/- 1.7%) and mild (100%) asthmatic tissues were identified as eosinophils. In contrast, eosinophils constituted a small portion of positive cells (20.8 +/- 21.6%) in normal tissues. These results indicated that TGF beta 1 mRNA was overexpressed in severe asthmatics and that the main source of the mRNA was eosinophils, suggesting that eosinophils play an important role in the pathogenesis not only of inflammation but also of structural changes, such as subepithelial fibrosis, in asthmatic airways.

Adult↗
PubMed1996

Molecular cloning and nucleotide sequences of cDNA and gene encoding endo-inulinase from Penicillium purpurogenum.

A cDNA and a gene encoding endo-inulinase from Penicillium purpurogenum were isolated, and were cloned for the first time. Two oligonucleotide probes, which were synthesized based on the partial amino acid sequences of the purified endo-inulinase, were used to screen a cDNA library. A 1.7-kb DNA fragment encoding endo-inulinase was isolated and analyzed. A single open reading frame, consisting of 1548-bp, was found to encode a polypeptide that comprised a 25-amino acid signal peptide and 490-amino acid mature protein. All the partial amino acid sequences of the purified enzyme were discovered in the deduced ones. The deduced amino acid sequences of endo-inulinase had similar sequences to those of fructan hydrolases. A 3.5-kb chromosomal DNA fragment encoding endo-inulinase was also isolated and analyzed. The same ORF with cDNA clone as identified. There were no introns in the endo-inulinase gene.

Amino Acid Sequence↗
PubMed1996

[The effect of new quinolone antimicrobial agents and macrolide antibiotics on the clearance of theophylline].

Many drugs have been found to increase or decrease the clearance of theophylline. Some of new quinolone antimicrobial agents (ciprofloxacin, enoxacin, pefloxacin and tosufloxacin) and macrolide antibiotics (erythromycin, troleandomycin, roxithromycin and clarithromycin), are potent inhibitors of the metabolism of theophylline. Concomitant administration of these drugs may, thus, prolong the half-life of theophylline, elevate serum theophylline concentrations, and increase the risk of theophylline-related adverse events. Therefore, these data indicate that careful clinical and pharmacokinetic monitoring of patients receiving both theophylline and some of antimicrobial agents should be carried out.

4-Quinolones↗
PubMed1995

Sudden hemorrhage of the breast caused by breast cancer without skin invasion: report of a case.

A rare case of sudden hemorrhage caused by breast cancer is herein presented. A 65-year-old woman was admitted to our hospital because of bleeding from her left breast. On physical examination, a continuous hemorrhage accompanied by an open cavity formation was observed in her left breast. She had no history of breast trauma. Her bleeding profile studies and liver function were both normal. In addition, no anticoagulation medication had been administered. The histological findings of the cavity wall indicated invasive ductal carcinoma without skin invasion. After carrying out chemotherapy, a standard radical mastectomy was performed.

Aged↗
PubMed1995

Establishment and characterization of a new cell line from human bladder cancer (JMSU1).

A new human bladder cancer cell line designated JMSU1 has been established from malignant ascitic fluid of a 75-year-old Japanese man with bladder cancer, and maintained in culture for more than 7 years and over 240 passages. Inverted phase-contrast microscopy revealed that JMSU1 was composed of morphologically distinct cells (polygonal to spindle-shaped cells), showing morphological heterogeneity in vitro. Histological examination of xenografts showed poorly differentiated transitional cell carcinoma, resembling the original tumor. Immunohistochemical staining for cytokeratin and electron microscopic examination suggested that JMSU1 was of epithelial origin. Chromosome analysis gave a modal number of 69 with no Y chromosome. Isozyme analysis (LDH, G6PD, and NP) showed the mobility pattern of human type B. DNA fingerprint analysis demonstrated that there was no cross-culture contamination of JMSU1 during the passages. In conclusion, a newly established and well-characterized cell line, JMSU1, offers promising material for the investigation of the biological properties of bladder cancer.

Aged↗
PubMed1995

A comparative study of TK6 human lymphoblastoid and L5178Y mouse lymphoma cell lines in the in vitro micronucleus test.

Micronucleus induction was compared in human lymphoblastoid TK6 and mouse lymphoma L5178Y cell lines treated with model clastogens and spindle poisons, i.e., X-rays, methyl methanesulfonate, ethyl methanesulfonate, mitomycin C, colcemid, and vincristine. The spontaneous micronucleated cell (MNC) frequency was stable and reproducible in both cell lines. All clastogens and spindle poisons studied here induced micronuclei in both cell lines. They increased MNC frequency at lower concentrations or caused a greater increase at the same concentration in TK6 cells. These clastogens and spindle poisons, however, were also more toxic to TK6 than to L5178Y cells and when comparison was based on cytotoxicity, they showed more efficient MNC induction in L5178Y cells. In conclusion, neither cell line was superior to the other, and both of them can be used as target cells in the in vitro micronucleus assay.

Animals↗
PubMed1995

DNA fingerprint analysis in chemically mutagenized Chinese hamster lung cells.

Using a multi-locus minisatellite Per-6 DNA probe, we performed DNA fingerprint analysis. Chinese hamster lung (CHL) cells were treated with six model chemicals: N-methyl-N'-nitro-N-nitrosoguanidine, mitomycin C, methyl methanesulfonate, furylfuramide, 2-acetylamino-fluorene, and cyclophosphamide, with or without S9 mix. 771 hypoxanthine phosphoribosyltransferase deficient clones (749 from mutagen-treated cells and 22 from untreated cells) and 90 unselected clones from untreated cells were isolated and analyzed. The spontaneous mutation frequency at CHL cell minisatellite loci was 0.31-0.63%. All the chemicals increased mutation frequencies. Almost all mutations localized to the three specific minisatellite loci corresponding to 4.2, 3.8, and 2.4 kb bands, suggesting that these regions are more unstable and susceptible to mutation. DNA fingerprint analysis is a promising technique for detecting mutations at neutral DNA regions, especially recombinational mutations, and may be useful for surveying genetic instability related to heritable defects or aging.

2-Acetylaminofluorene↗
PubMed1995

Recombinagenic activity of the phorbol ester 12-O-tetradecanoylphorbol-13-acetate in human lymphoblastoid cells.

The potent mouse skin tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA) was examined for its mutagenic and recombinagenic activity at the heterozygous thymidine kinase (tk +/-) locus and the hemizygous hypoxanthine phosphoribosyltransferase (hprt +/0) locus in the TK6 human lymphoblastoid cell line. TPA at concentrations of 0.01-1.0 micrograms/ml induced a low frequency of tk mutants showing the slow growth phenotype in a dose-dependent manner, but few normal growth tk mutants or hprt mutants. Concentrations of 1.0-10 micrograms/ml TPA induced all three types of mutants. The molecular structure of tk mutants arising spontaneously or induced by 1.0 and 10 micrograms/ml TPA was investigated by Southern hybridization with a human tk cDNA probe: 86% of all mutants arising after incubation with 10 micrograms/ml TPA lost the entire active tk allele, resulting in loss of heterozygosity (LOH), while 71% of spontaneously arising mutants showed LOH. Densitometric analysis indicated that the majority of LOH mutants induced by TPA were homozygous at the tk locus (retained two copies of the mutant allele), consistent with the occurrence of interchromosomal homologous recombination. These results support the hypothesis that tumor promoters such as TPA may increase the rate of chromosomal mitotic recombination and hence facilitate the segregation of recessive mutations. TPA may thus induce a type of genetic instability during the process of tumor promotion that involves enhanced recombinagenic activity.

Cell Line↗
PubMed1995

Combination effects of clastogens in the mouse peripheral blood micronucleus assay.

In order to study how two chemicals interact to induce micronuclei, simple ethylating agents [ethyl methanesulfonate (EMS), ethyl ethanesulfonate (EES) and N-ethyl-N-nitrosourea (ENU)], spindle poisons [vincristine sulfate (VINC) and colchicine (COL)] and an oxidizing agent [potassium bromate (KBrO3)] were used as model chemicals for combination treatments. The frequency of micronucleated reticulocytes (MNRETs) was evaluated in mice treated with two of these chemicals at a time. The combinations of ethylating agents (EMS and EES; EMS and ENU) and of spindle poisons (VINC and COL) induced more micronuclei than those expected on an additive basis. The apparent synergism was due to a 'combined dose' which could be calculated by the dosimetric conversion of one chemical to the other, when damage induced by each chemical was 'equivalent' in the induction of MNRETs. In contrast, no apparent synergism in induction of micronuclei was observed when two chemicals with different modes of clastogenic action (EMS and KBrO3 or EMS and VINC) were combined.

Alkylating Agents↗
PubMed1995

Analysis by fluorescence in situ hybridization with a mouse gamma satellite DNA probe of isolated micronuclei induced in mice by two clastogens and two spindle poisons.

The presence of centromeric DNA was studied in micronuclei isolated from the blood of male ddY mice after five weekly intraperitoneal injections of mitomycin C (MMC), 1-beta-D-arabinofuranosylcytosine (Ara-C), colchicine (COL) or vinblastine sulfate (VBL). In agreement with our earlier findings, about half of the micronuclei isolated from vehicle control mice showed centromere signals as analyzed by fluorescence in situ hybridization (FISH) with a mouse major (gamma) satellite DNA probe. In an earlier experiment with mice acutely exposed to the same chemicals, the clastogens MMC and Ara-C did not reduce the proportion of micronuclei with centromere signals. In the present study, however, MMC and Ara-C decreased the proportion of micronuclei with centromeres. In contrast, the spindle poisons COL and VBL increased the proportion of micronuclei that contained centromeres.

Animals↗
PubMed1995

IL-5, IL-8 and GM-CSF immunostaining of sputum cells in bronchial asthma and chronic bronchitis.

BACKGROUND: Granulocyte-macrophage colony stimulating factor (GM-CSF) and interleukin (IL)-5 or IL-8 have been suggested to play an important role in the pathogenesis of eosinophilic airway inflammation in bronchial asthma or neutrophilic airway inflammation in chronic bronchitis, respectively, However, GM-CSF and IL-8 have biological activities to either eosinophils or neutrophils. OBJECTIVE: To investigate the contribution of these cytokines to airway inflammation, we compared the cellular differential and immunolocalization of GM-CSF, IL-5 and IL-8 in sputum cells from patients with bronchial asthma and chronic bronchitis. METHODS: Cytospins of sputum cells from 12 patients with bronchial asthma and 12 with chronic bronchitis were subjected to cellular differential counting and immuno-cytochemistry with anti-human GM-CSF, IL-5 and IL-8 antibody. RESULTS: The predominant cells in bronchial asthma were eosinophils and lymphocytes, while those in chronic bronchitis were neutrophils. All cytokines examined were detected in either bronchial asthma or chronic bronchitis, although the percentage of GM-CSF and Il-5 positive cells in bronchial asthma (53.4 +/- 6.0 [mean +/- SEM]% and 9.7 +/- 2.8%, respectively) was significantly higher than that in chronic bronchitis (11.4 +/- 2.5%; P < 0.001 and 1.7 +/- 0.3%; P < 0.007, respectively). In contrast, the percentage of IL-8 positive cells in chronic bronchitis (23.8 +/- 7.0%) was significantly higher than that in bronchial asthma (7.& +/- 1.9%; P < 0.04). The cells positive for IL-5 were lymphocytes in bronchial asthma and chronic bronchitis. The cells positive for GM-CSF in bronchial asthma were predominantly eosinophils, while those in chronic bronchitis were monocytes/macrophages and neutrophils. In contrast, neutrophils are mainly positive for IL-8 in chronic bronchitis, while monocytes/macrophages and bronchial epithelial cells are positive for IL-8 in bronchial asthma. CONCLUSION: The immunochemical comparison of GM-CSF and IL-8 localization in sputum cells between bronchial asthma/chronic bronchitis suggests the differential regulation and roles of these cytokines in eosinophilic vs neutrophilic airway inflammation, resulting in the development of different types of airway inflammation.

Adult↗
PubMed1995

Co-expression of fibroblastic, histiocytic and smooth muscle cell phenotypes on cultured adherent cells derived from human palatine tonsils: a morphological and immunocytochemical study.

Adherent cells derived from human palatine tonsils were isolated and cultivated. Exponentially growing adherent cells (TAC) were observed by phase-contrast microscopy and transmission electron microscopy. Immunocytochemical studies were also performed. TAC were composed of relatively monotonous cells with polygonal or spindle shapes and high proliferative activity. In addition to the development of rough endoplasmic reticulum and lysosomes, the TAC possessed a moderate amount of pinocytotic vesicles and a few microfilaments. All of the TAC strongly expressed fibroblastic markers and partial monocyte/macrophage markers, such as beta-subunit of prolyl 4-hydroxylase (DAKO-fibroblast), lysozyme, anti-alpha-1-antichymotrypsin (alpha ACT), and CD68 (KP-1, EBM/11). It was noted that, as the TAC were cultured for a longer period, they gradually increased the reactivity with the monoclonal antibody PG-M1. Furthermore, the TAC expressed myocytic phenotype, such as alpha-smooth muscle actin (alpha SMA) with various intensity. Moreover, as to extracellular matrix, TAC stained for collagen type I, collagen type III, laminin, and fibronectin. Collagen type IV was weakly positive. The results presented here showed that the TAC expressed three different phenotypes of fibroblasts, histiocytes and smooth muscle cells at the same time. The monoclonal antibody raised against the TAC reacted strongly with the subendothelial pericytes and/or smooth muscle cells in the extrafollicular area in human tonsils. The present results also suggested that the origin of the TAC was probably subendothelial pericytes and/or smooth muscle cells of the microvasculatures in the tonsil.

Adult↗
PubMed1995

In situ expression of the cell adhesion molecules in bronchial tissues from asthmatics with air flow limitation: in vivo evidence of VCAM-1/VLA-4 interaction in selective eosinophil infiltration.

Eosinophils play a critical role in the pathogenesis of bronchial asthma by releasing various mediators. To understand the mechanisms of eosinophil migration to the site of inflammation, we examined the expression of adhesion molecules in the bronchial tissues of asthmatic subjects with air flow limitation. By immunohistochemical analysis, Mac-1, LFA-1, and VLA-4 were strongly positive in eosinophils and mononuclear cells infiltrated in the bronchial mucosa and submucosa. Their number was significantly increased compared with those in control tissue. Immunolocalization for ICAM-1, the ligand of Mac-1 and LFA-1, was detected in the endothelial cells of capillaries and venules, in the mononuclear cells in submucosa, and in the basal layer of the epithelium. Endothelial cells in capillaries and venules were also strongly positive for VCAM-1, the ligand of VLA-4. Immunolocalization for E-selectin was detected in some endothelial cells in capillaries and venules in bronchial submucosa, whereas there were very few positive cells in the bronchial tissues from control subjects. In situ hybridization demonstrated ICAM-1 mRNA expression in the endothelial cells and mononuclear cells in bronchial submucosa. Immunoelectron microscopy for ICAM-1, VCAM-1, and E-selectin demonstrated de novo synthesis of these molecules and their expression along the luminal cell membrane of endothelial cells. These results suggested that ICAM-1, VCAM-1, and E-selectin were newly synthesized prior to spontaneous asthma attacks, and that their expression, particularly that of VCAM-1, may play a key role in eosinophil infiltration into the airway.

Adult↗
PubMed1995

Eosinophils as a potential source of platelet-derived growth factor B-chain (PDGF-B) in nasal polyposis and bronchial asthma.

Tissue remodeling in bronchial tissues from asthmatics as well as in nasal polyp (NP) tissues includes sub-basement membrane deposition of collagen, stromal deposition of extracellular matrix protein, and hypertrophy/hyperplasia of airway smooth muscle cells, which are relevant to the cellular and molecular events induced by platelet-derived growth factor (PDGF). Therefore, we investigated the localization of mRNA and protein of PDGF-B chain (PDGF-B) in NP tissues and bronchial tissues from mild and severe asthmatics by in situ hybridization and immunohistochemistry, respectively. Cells expressing PDGF-B mRNA were found in all nine NP tissues and in bronchial tissues from 2 of 6 normal subjects, 2 of 5 mild asthmatics, and all of 6 severe asthmatics examined. The vast majority of cells expressing PDGF-B mRNA were eosinophils in NP (99.7 +/- 0.2%, mean +/- SD) and asthmatic bronchial tissues (75.0 and 77.8% in mild asthma, and 92.7 +/- 8.1% in severe asthma), but no cells expressing PDGF-B mRNA were eosinophils in normal bronchial tissues. The number of cells expressing the gene in severe asthma tissues (122.3 +/- 32.2/mm2) was similar to that in NP tissues (152.8 +/- 73.9/mm2) and greater than that in mild asthma tissues (4.7 +/- 7.6/mm2, P < 0.01), which was not significantly greater than that in normal bronchial tissues (3.4 +/- 5.2/mm2). Furthermore, we detected immunolocalization of PDGF-B in NP tissues and in asthmatic bronchial tissues. The eosinophils purified from peripheral blood were demonstrated to express PDGF-B gene transcript and immunoreactivity after stimulation with A23187.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1995

Chemical modification and amino acid sequence of active site in sugar beet alpha-glucosidase.

The modification of amino acid residues in sugar beet alpha-glucosidase with conduritol B epoxide (CBE), an affinity labeling reagent, inactivated the enzyme. The inactivation followed pseudo-first-order kinetics. The enzyme was protected from inactivation by a competitive inhibitor, Tris, and the partially inactivated enzymes showed only the decrease of V values and no change in Km value. An 3H-CBE labeled peptide isolated from the digest of the inactivated enzyme with Lys-C protease was sequenced. The -COO- group of Asp was found to be specifically labeled, implicating that it is a catalytic group of the enzyme. The sequence around the essential Asp was determined to be -DGIWIDMNE-, which showed a high homology with those of other alpha-glucosidases.

Amino Acid Sequence↗
PubMed1994

Isolation of micronuclei from mouse blood and fluorescence in situ hybridization with a mouse centromeric DNA probe.

Spontaneously existing and chemically induced micronuclei were isolated from mouse blood. 50 microliters of cardiac blood was diluted with PBS and centrifuged. After this, the cell pellet was subjected to hypotonic treatment, fixed with acetic acid-methanol (1:3), and the lysate was filtrated through a 2-microns polycarbonate nucleopore membrane. Isolated micronuclei were air-dried on a glass slide and subjected to fluorescence in situ hybridization (FISH) using a mouse centromeric gamma satellite probe. Approximately half of the micronuclei isolated from vehicle control mice showed centromere signal(s). In these preliminary studies, the proportion of centromere-positive micronuclei was increased by treatment with spindle poisons (colchicine and vinblastine sulfate), decreased only slightly by 1-beta-D-arabinofuranosylcytosine, and was generally unaffected by mitomycin C.

Animals↗