Search PubMed⌕ Search

Biomedical subjects

Z Honda

Publications and source records attributed to Z Honda.

46 records · Page 3Linked to original sources
PubMed1988

Solubilization and partial purification of leukotriene C4 synthase from guinea-pig lung: a microsomal enzyme with high specificity towards 5,6-epoxide leukotriene A4.

Enzymic activities catalyzing allylic epoxide, leukotriene A4, to leukotriene C4 by conjugation with glutathione were present mainly in microsomal fractions of spleens and lungs of guinea pigs and rats. Leukotriene C4 (LTC4) synthase was solubilized from the microsomes of guinea-pig lung by the new procedures of a combination of 3-[3-cholamidopropyl)dimethylammonio)-1-propanesulfonate (CHAPS), digitonin and KCl. The enzyme was partially purified by two steps of column chromatography which resulted in a complete resolution of the enzyme from glutathione S-transferases (EC 2.5.1.18). The partially purified LTC4 synthase showed a Vmax value of 40 nmol/min per mg, and the apparent Km values for LTA4 and glutathione were 36 microM and 1.6 mM, respectively. The enzyme was unstable, and half of the activity was lost by incubation at 37 degrees C for 3 min. Glutathione at 10 mM completely protected the enzyme against this inactivation, while other sulfhydryl-group-reducing reagents were ineffective. The partially purified enzyme revealed a high specificity towards 5,6-epoxide leukotrienes (LTA4 and its methyl ester), while rat cytosolic glutathione S-transferases catalyzed conjugation of glutathione to various positional isomers of epoxide leukotrienes.

Animals↗
PubMed1988

Role of protein kinase C in histamine release from human basophils.

In this study, we investigated the role of calcium and phospholipid-dependent protein kinase (protein kinase C, PKC) in the modulation of histamine release from human basophils. A novel and potent inhibitor of PKC, K-252a, inhibited the release of histamine induced by anti-IgE in a dose-dependent manner with ID50 (the dose required for 50% inhibition of histamine release) of 2.2 x 10(-8) M. Histamine release stimulated with 12-0-tetradecanoyl-phorbol-13-acetate(TPA) was also suppressed by K-252a with maximal inhibition of 48.0 +/- 9.3% at 10(-7) M. In contrast, K-252a did not inhibit the release of histamine in response to FMLP and ionophore A23187. Another inhibitor of PKC, H-7, exhibited a dose-dependent inhibition of anti-IgE-induced histamine release with ID50 of 8.6 x 10(-4) M. H-8 and HA1004, which closely resemble H-7 in chemical structure but are less potent in inhibiting PKC, did not inhibit histamine release stimulated with anti-IgE, but rather enhanced the release at higher concentrations. These results strongly suggest that PKC activation plays a crucial role in the mediation of IgE-mediated histamine release from human basophils.

Antibodies, Anti-Idiotypic↗
PubMed1987

Differential inhibitory effects of auranofin on leukotriene B4 and leukotriene C4 formation by human polymorphonuclear leukocytes.

Auranofin (AF) is a newly introduced oral gold compound having antirheumatic properties, and its efficacy in the treatment of bronchial asthma is now under investigation. In this study, we examined the effects of AF on leukotriene (LT) formation by human polymorphonuclear leukocytes (PMNs) stimulated with the calcium ionophore A23187. AF inhibited LTC4 formation in a dose-dependent manner with an IC50 (concentration required to produce 50% inhibition of control) of 3.2 microM. In contrast, LTB4 formation was not prevented by AF at concentrations up to 6 microM, but it was reduced to 59 +/- 4% (mean +/- SE, N = 3) of control by an 8 microM concentration. As a next step, we explored the mechanisms of the differential inhibitory effects of AF using cell-free systems. When arachidonic acid (AA) and reduced glutathione (GSH) were used as substrates, AF inhibited LTC4 synthesis more effectively (IC50 = 14 microM) than LTB4 synthesis (IC50 = 100 microM). However, LTB4 and LTC4 syntheses from LTA4 were affected only slightly by AF within the concentrations tested (3-100 microM). These results in the cell-free systems indicate that the inhibition of LT formation was caused by a reduction of LTA4 synthesis and that the differential inhibitory effects can be ascribed to the higher Km value of glutathione S-transferase for LTA4 than that of LTA4 hydrolase in PMNs. In accordance with this hypothesis, LTC4 synthesis was more dependent than LTB4 synthesis on LTA4 concentrations within 25-100 microM, and AA-861, a 5-lipoxygenase inhibitor, caused similar differential inhibitory effects on the formation of LTs by intact PMNs. The inhibitory effect of AF on LT formation at physiological concentrations may play some role in the efficacy of this drug.

Animals↗
PubMed1987

Clinical evaluation of three types of plasmapheresis in a patient with type IIa familial hypercholesterolemia.

Long-term plasmapheresis (PP) therapy was studied in a 56-year-old patient with homozygous type IIa familial hypercholesterolemia also suffering from severe coronary heart disease. Three different PP techniques, plasma exchange (PE), double-membrane-filtration plasmapheresis (DFP), and the recently developed low-density lipoprotein adsorbent column (adsorption plasmapheresis, adsorption PP), were used in an attempt to develop better means of managing the disease. All three techniques showed similar elimination efficiency with respect to plasma total cholesterol level. Adsorption PP with minimal supplemental plasma protein managed the circulatory status of the patient better than DFP during extracorporeal treatment. In the course of PP therapy xanthoma tuberosum markedly regressed, and the cardiac status of the patient was clearly improved.

Angiography↗
PubMed1987

Suppression of human polymorphonuclear leukocyte phagocytosis by adenosine analogs.

The effects of adenosine analogs on human polymorphonuclear leukocyte (PMN) phagocytosis to latex beads or sheep red blood cells were investigated in an in vitro experiment. The purine compounds such as N6-phenylisopropyladenosine (PIA), 5'-N-ethylcarboxamido-adenosine (NECA), an A2 adenosine receptor agonist, and adenosine as high as 2 mM had no effect on PMN phagocytosis. In contrast, 2',5'-dideoxyadenosine (DDA), a P-site adenosine receptor agonist, and 5'-methylthioadenosine (MTA), a naturally occurring purine nucleoside, caused profound inhibition of phagocytic function of PMNs in a dose-dependent manner. Dipyridamole, which blocks purine nucleoside uptake, reversed the suppression due to MTA. Theophylline, an R-site receptor antagonists did not prevent the effect of MTA. These results suggested that phagocytosis of PMNs is suppressed by stimulation of the P-site adenosine receptors.

Adenosine↗
PubMed1986

Differential composition of cytosol 5'-nucleotidases between T and B lymphoblasts.

WI-L2 cells (a B-lymphoblastoid cell line) were more resistant than CEM cells (a T-lymphoblastoid cell line) to deoxyadenosine, ara-A (9-beta-D-arabinofuranosyladenine), or ara-C (1-beta-D-arabinofuranosylcytosine) inhibition. This was caused by a difference in the composition of cytosol 5'-nucleotidases between WI-L2 and CEM cells. In intact cells, the endogenous production of deoxyadenosine from WI-L2 cells deficient in adenosine kinase (EC 2.7.1.20) and deoxycytidine kinase (EC 2.7.1.74) was consistently high, despite changes in endogenous adenosine production. Endogenous production of deoxyadenosine from CEM cells deficient in adenosine kinase and deoxycytidine kinase was, however, coordinated with endogenous adenosine production. In broken cells, cytosol dAMPase (2'-deoxyadenosine 5'-monophosphate 5'-nucleotidase) activity of WI-L2 cells was 3-5-fold higher than that of CEM cells. dAMPase activity could be separated from ATP-activated IMPase (inosine 5'-monophosphate 5'-nucleotidase) by gel filtration (molecular weight: dAMPase; 39,000-46,000; ATP-activated IMPase, greater than 150,000). Cytosol ATP-activated IMPase and dAMPase were isolated by phosphocellulose or DEAE-Bio-Gel A chromatography from non-specific phosphatases. The ATP-activated IMPase showed only marginal activity towards dAMP (2'-deoxyadenosine 5'-monophosphate), ara-AMP (9-beta-D-arabinofuranosyladenine 5'-monophosphate), or ara-CMP (cytosine-beta-D-arabinofuranoside 5'-monophosphate), even in the presence of ATP. The activity of ATP-activated IMPase was similar in WI-L2 and CEM cells. dAMPase was separated into two peaks by DEAE-Bio-Gel A chromatography; one of these peaks degraded ara-AMP and ara-CMP. The activities of both peaks from WI-L2 cells were higher than those from CEM cells. These results show that the degradation of dAMP, ara-AMP or ara-CMP was more specific and rapid in WI-L2 than in CEM cells.

5'-Nucleotidase↗
PubMedDate not supplied

Properties of the guinea-pig lung platelet-activating factor receptor encoded by the cloned cDNA.

Platelet-activating factor (PAF) is a naturally occurring phospholipid possessing proinflammatory and shock-inducing activities. Although PAF exerts these functions via specific receptors, the molecular structures of the receptor has not been clarified. We have isolated a cDNA coding for PAF receptor through expression cloning. The PAF receptor expressed in Xenopus laevis oocytes and COS-7 cells showed pharmacological properties consistent with those in previous reports. Sequence analysis revealed that PAF receptor belongs to the superfamily of G-protein-coupled receptors.

Animals↗
PubMedDate not supplied

Isolation and properties of platelet-activating factor receptor cDNAs.

The cDNA encoding the platelet-activating factor (PAF) receptor was cloned from a guinea pig lung cDNA library by using a Xenopus laevis oocyte expression system. The human PAF receptor cDNA was isolated from a human leukocyte cDNA library using a 0.8-kbp fragment of the guinea pig PAF receptor cDNA as a probe. Both receptors have the same number of amino acids (342 residues) with seven putative transmembrane spanning domains, and belong to the G-protein-linked receptor superfamily. Overall amino acid identity between the two receptors was 83%:91% in the transmembrane domains. Seven threonine and three serine residues conserved in the cytoplasmic loops of both receptors may function as phosphate acceptors, as related to the homologous desensitization of the receptor. Activation of the PAF receptor yielded inositol 1,4,5-trisphosphate production in the PAF receptor expressed COS-7 cells and oocytes, and guanosine 5'-O-(2-thio)bisphosphate injected into the oocytes inhibited PAF-induced Cl- current, providing evidence that PAF stimulates phosphoinositide turnover via G-protein(s).

Amino Acid Sequence↗