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ABSTRACT: Rat peritoneal mast cells were incubated with different concentrations of naturally occurring aliphatic polyamines, spermine and spermidine, at 0.1-10 mM and the amount of histamine release into the supernatant solutions was measured. The addition of each polyamine to the suspensions of the mast cells caused a histamine release in a dose-dependent manner. The effect of 10 mM spermine and spermidine was as much as that of 0.5 microgram/ml compound 48/80. The histamine release from the cells incubated with each polyamine was rapid and the amount of histamine release into the supernatant solutions reached a maximum at 1 min with the incubations. 0.1 mM spermine, which in itself could not cause a significant histamine release, showed a tendency to enhance anti-IgE-induced histamine release from the mast cells.
Allergy 08/1991; 46(5):349-54. · 6.27 Impact Factor
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ABSTRACT: Blood was drawn from healthy human volunteers and a 30-year-old woman with severe bronchial asthma (atopic and perennial type), whose peripheral blood platelets counts were continuously higher than 60 x 10(4)/mm3. Neutrophils were purified on a Conray-Ficoll gradient and platelets were isolated by a gel filtration method with Sepharose 2B. Superoxide anion (O2-) generation from the cells was measured by 2-methyl-6-[p-methoxyphenyl]-3,7-dihydroimidazo [1,2-a]purazin-3-one (MCLA)-dependent luminescence. Addition of 0.5 microM MCLA and 10(-8) M N-formyl-methionyl-leucyl-phenylalanine (fMLP) or 10 ng/ml phorbol myristate acetate (PMA) to a suspension of platelets obtained from the healthy controls and the patient caused no significant MCLA-dependent luminescence, 02- generation by neutrophils from healthy human volunteers, activated by 10(-8) M fMLP or 10 ng/ml PMA, was inhibited by their platelets in a concentration-dependent manner. Platelets from the patient failed to inhibit O2- generation from neutrophils activated by 10(-8)M fMLP. Cu-Zn superoxide dismutase concentrations in platelets from healthy human volunteers were not higher than the patient's. These data suggest that the absence of inhibition of superoxide generation from activated neutrophils by platelets might be involved in the pathogenesis of bronchial asthma.
Allergy 05/1991; 46(3):173-9. · 6.27 Impact Factor
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ABSTRACT: To investigate the possible influences of hyperthyroidism on the pathophysiology of bronchial asthma, we clinically observed the effect of antihyperthyroidal therapy on asthmatic symptoms in 7 patients who shared both disorders. In five cases, asthma symptoms were affected by the therapy. Asthma symptoms improved after normalization of thyroid function in two cases, but worsened after therapy for hyperthyroidism in three cases. There was no apparent correlation between the pathophysiology of the two diseases after therapy in the two other cases. These results showed that thyrotoxicosis does not uniformly affect the pathophysiology of asthma and emphasizes the need to follow asthmatic patients closely following treatment for hyperthyroidism.
Journal of Asthma 02/1991; 28(2):109-16. · 1.52 Impact Factor
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ABSTRACT: U937 cells are established human monoblast cell line originally isolated from a patient with diffuse histiocytic lymphoma. In the present study, superoxide anion (O2-) generation from U937 cells were measured by 2-methyl-6-[p-methoxyphenyl]-3,7- dihydroimidazo[1,2-a]pyrazin-3-one (MCLA)-dependent luminescence. Addition of 0.5 mumol/l MCLA and a stimulatory agent, such as N-formyl-methionyl-leucyl- phenylalanine (fMLP) and phorbol myristate acetate (PMA), to a suspension of the cells treated with gamma interferon (IFN gamma) caused a marked luminescence which was inhibited by 0.5 mumol/l superoxide dismutase (SOD). On the other hand, the cells which were not treated with IFN gamma caused no significant luminescence by the stimulation with either fMLP or PMA. The adherent IFN gamma-treated cells generated more O2- than the nonadherent IFN gamma-treated cells by the stimulation with fMLP. The maximal luminescence intensity from the adherent IFN gamma-treated U937 cells stimulated by fMLP was dependent upon the number of the cells. Azelastine (A-5610, CAS 58581-89-8) significantly inhibited the O2- generation from the adherent IFN gamma-treated U937 cells stimulated by fMLP or PMA in a dose-dependent manner.
Arzneimittel-Forschung 02/1991; 41(1):43-7. · 0.72 Impact Factor
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ABSTRACT: Human alveolar macrophages were recovered by bronchoalveolar lavage and superoxide anion (O2-) generation from the macrophages were measured by 2-methyl-6-[p-methoxyphenyl]-3,7-dihydroimidazo[1,2-a]pyrazin++ +-3-one (MCLA)-dependent luminescence. Addition of 0.5 mumol/l MCLA and a stimulatory agent, such as phorbol myristate acetate (PMA) and N-formyl-methionyl-leucyl-phenylalanine (fMLP), to a suspension of the cells caused a marked luminescence which was inhibited by 0.5 mumol/l superoxide dismutase (SOD). The maximal luminescence intensity was dependent upon the number of the cells. Azelastine (A-5610, CAS 58581-89-8) significantly inhibited the O2- generation from the activated macrophages in a dose-dependent manner. However, the other antiallergic drugs, such as ketotifen, disodium cromoglycate and others showed little effects on the O2- generation from the activated human alveolar macrophages.
Arzneimittel-Forschung 02/1991; 41(1):47-51. · 0.72 Impact Factor
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ABSTRACT: Blood was drawn from healthy human volunteers and neutrophils and eosinophils were purified on a Conray-Ficoll and a Percoll gradient, respectively. Rat mast cells were also purified on a Percoll gradient. Superoxide anion (O2-) generation from the cells were measured by 2-methyl-6-[p-methoxy-phenyl]-3,7-dihydroimidazo[1,2-a]pyrazin+ ++-3-one (MCLA)-dependent luminescence. Addition of 0.5 mumol/l MCLA and a stimulatory agent, such as phorbol myristate acetate, N-formyl-methionyl-leucyl-phenylalanine (fMLP) and compound 48/80, to a suspension of each cell caused a marked luminescence which was inhibited by 0.5 mumol/l superoxide dismutase (SOD). Azelastine (A-5610) significantly inhibited the O2- generation from each activated inflammatory cell in a dose-dependent manner. When eosinophils were activated by fMLP in the presence of cytochalasine (CB), azelastine abolished the luminescence stronger than that from the fMLP-stimulated cells in the absence of CB.
Arzneimittel-Forschung 08/1990; 40(7):767-70. · 0.72 Impact Factor
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ABSTRACT: We report here an efficient method for simultaneous and quantitative determinations of plasma prostaglandins (PGs) from asthmatic patients using 9-anthryldiazomethane-reversed-phase high performance liquid chromatography (HPLC). Detection and simultaneous quantitation of 20 pg PGE1, PGE2, PGF2 alpha and 100 pg 6-keto PGF1 alpha, thromboxane B2 (TXB2) was possible by the present method. Preliminary studies showed that plasma TXB2 levels from stable asthmatic patients were elevated compared with normal healthy subjects, suggesting pathophysiologic implications regarding PGs and TXB2 in asthma may be elicited by the present HPLC method.
Annals of allergy 06/1990; 64(5):464-70.
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ABSTRACT: Rat mast cell granules were obtained by homogenization of highly purified rat mast cells and isolated in a Percoll gradient. Diphosphoinositide (DPI) synthesis in rat mast cell granules was assayed by measuring the incorporation of 32P from [gamma 32P] ATP into DPI in the absence of exogenous phosphatidylinositol (PI). Lipids were isolated with methanol/chloroform/HCl and were separated by thin-layer chromatography on oxalic acid impregnated silica gel plates. DPI areas were identified by staining with iodine, scraped and measured for 32P radioactivity. The addition of polyamines, spermine and spermidine, to the granules caused an increase of DPI synthesis, which can be catalyzed by PI kinase. This effect of polyamines in the DPI synthesis was in a dose-dependent manner and maximal effects were observed at 1 mM spermine and 10 mM spermidine, respectively.
Allergy 06/1990; 45(4):262-7. · 6.27 Impact Factor
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ABSTRACT: To determine why midaglizole is effective in some patients with severe asthma, we investigated the inhibitory effects of midaglizole, prazosin or yohimbine on the BHT 920-, phenylephrine-, or noradrenaline-induced contractions of canine tracheal smooth muscle. After pretreatment with atropine (10(-6) M) and propranolol (10(-6) M) and precontraction with serotonin (3 x 10(-7) M), the tracheal muscle showed contractile responses to the exogenous administration of both alpha 1 and alpha 2 adrenoceptor agonists. Every alpha antagonist inhibited these agonist-induced contractions. Inhibitory activity of midaglizole (10(-4) M) for the alpha agonists was BHT-920 greater than noradrenaline greater than or equal to phenylephrine, while that of prazosin (3 x 10(-6) M) was phenylephrine greater than noradrenaline greater than BHT-920. Moreover, yohimbine completely inhibited the contractions at the lower concentration of 3 x 10(-7) M than that of other two antagonists. Our findings demonstrate that midaglizole dose-dependently inhibits airway contractions induced by alpha adrenoceptor agonists.
Arerugī = [Allergy] 05/1990; 39(4):384-90.
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ABSTRACT: Histamine release from peripheral leukocytes of "Konnyaku asthmatic patients," patients with a unique occupational bronchial asthma in Japan, was measured after challenge with the antigen. There was an antigen concentration-dependent response with highly significant correlations between the maximal histamine release and the ratio of the radioallergosorbent test (RAST) of the antigen to human serum albumin. These results suggest that the histamine release test from peripheral leukocytes as well as RAST can be a useful diagnostic test in patients with bronchial asthma.
Annals of allergy 04/1990; 64(3):319-21.
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ABSTRACT: Rat mast cell granules and plasma membrane fractions were obtained by homogenization of highly purified rat mast cells and isolation in a Percoll gradient and a sucrose gradient, respectively. Immunostaining of rat mast cells, granules and plasma membrane fractions was performed with mouse monoclonal antibody M6764 which was produced against the crude membrane fractions of the neural tubes. Rat mast cells and granules were immunostained with the monoclonal antibody, but not the plasma membrane fractions. The granules fixed with glutaraldehyde-paraformaldehyde showed ring-like forms. Chloroform-methanol treatment did not effect the staining of rat mast cells and granules with the monoclonal antibody. Western blotting analysis of rat mast cells and granules with the monoclonal antibody showed broad protein bands ranging from 100 to 250 kD.
International archives of allergy and applied immunology 02/1990; 92(4):349-55.
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ABSTRACT: Plasma prostaglandin (PG) and thromboxane (Tx) levels from bronchial asthmatic patients were assayed by 9-anthryldiazomethane reversed-phase high-performance liquid chromatography method. TxB2 levels from stable bronchial asthmatic patients were significantly higher than those from normal healthy subjects. PGF2 alpha levels were significantly elevated in atopic patients compared with nonatopic ones. PG patterns differed between asthmatic and nonasthmatic conditions in the same patients. PGF2 alpha levels were significantly elevated in the patients with spontaneous attacks compared with those without attacks and 6-keto PGF1 alpha levels were significantly lower in bronchial asthmatic patients having attacks than in the patients without attacks. There were no significant differences of PG levels between mild and moderate asthmatic subjects. Aminophylline injection in patients with mild spontaneous attacks had no significant effects on PG levels which were compared before and after injection.
Journal of Asthma 02/1990; 27(6):349-58. · 1.52 Impact Factor
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ABSTRACT: Rat mast cell granules were obtained by sonication of highly purified rat mast cells and isolated in a Percoll gradient. Phosphorylation of endogenous phosphatidylinositol in rat mast cell granules, which is catalyzed by phosphatidylinositol kinase in the granules, was assayed by measuring the incorporation of 32P from [gamma 32P]ATP into phosphatidylinositol 4-phosphate. Lipids were isolated with methanol/chloroform/HCl and were separated by thin-layer chromatography on oxalic acid impregnated silica gel plates. Phosphatidylinositol 4-phosphate areas were identified by staining with iodine, scraped and measured for 32P radioactivity. The phosphorylation reaction was inhibited by 50-500 microM adenosine, ADP and 500 microM AMP in a concentration-dependent manner. Among several anti-allergic drugs investigated. 100-1000 microM theophylline and 10-100 microM azelastine inhibited the phosphorylation reaction, but disodium cromoglycate and ketotifen had little effect.
Allergy 12/1989; 44(8):576-81. · 6.27 Impact Factor
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ABSTRACT: Rat serosal mast cells were challenged with compound 48/80 or calcium ionophore A23187 and the effect of staurosporine, a new inhibitor of protein kinase C, on histamine release from the cells was investigated. Histamine release induced by compound 48/80 or calcium ionophore A23187 was inhibited by staurosporine in a concentration-dependent manner and 0.1 and 1 microM staurosporine inhibited the histamine release significantly. The inhibitory effect of K-252a, another novel protein kinase C-inhibitor, was significantly higher than that of staurosporine on calcium ionophore A23187-induced histamine release. These results suggest that protein kinases will be involved in the process during mediator release from rat serosal mast cells.
Annals of allergy 10/1989; 63(3):231-4.
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ABSTRACT: Intact granules were isolated from sonicated purified rat serosal mast cells on a Percoll gradient. The granules were demonstrated to contain a diphosphoinositide kinase that catalyzes the formation of triphosphoinositide from diphosphoinositide. The enzyme requires adenosine triphosphate and Mg2+ or Mn2+ for activity. The Km for adenosine triphosphate is 3 mumol/L, and maximal response is observed at 20 mmol/L of Mg2+ or 1 mmol/L of Mn2+, respectively. Triphosphoinositide synthesis in the granules is dependent on the time and temperature of the incubations. Cyclic adenosine monophosphate, adenosine, adenosine diphosphate, and adenosine monophosphate decrease the enzyme activity. A comparison of the rate of phosphorylation of intact and broken membrane granules suggests that the phosphorylation occurs on the outer (cytoplasmic) surface of the granules.
Journal of Allergy and Clinical Immunology 08/1989; 84(1):118-28. · 11.00 Impact Factor
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ABSTRACT: Rat mast cells were challenged with compound 48/80 or calcium ionophore A23187, and protein kinase C activity in the cell pellets and the amount of histamine release into the supernatant were measured. After stopping the reaction, rat mast cells were lysed in a medium which prevents alterations in phosphorylation and dephosphorylation during sample processing. Histamine was significantly released from compound 48/80-stimulated mast cells at 30 s after the stimulation. In parallel with this, protein kinase C activity in the cell pellets increased at 30 s and 1 min and returned to basal value 3 min after the stimulation. When mast cells were incubated with various concentrations of 48/80 for 30 s, the amount of histamine release and protein kinase C activity increased dependently on the concentration of 48/80. Significant histamine release from A23187-stimulated mast cells was found at 1 min after the stimulation. Also protein kinase C activity in the cell pellets increased at 1 min and returned to basal value 5 min after the stimulation. A reduction of cytosolic protein kinase C activity was observed upon 48/80 treatment in a time- and concentration-dependent manner. Further, staurosporine, a potent inhibitor of protein kinase C, inhibited 48/80-induced histamine release in parallel with the inhibition of protein kinase C activity. These findings suggest that transient increase of protein kinase C activity may be involved in the mast cell activation process.
Allergy 05/1989; 44(3):226-32. · 6.27 Impact Factor
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ABSTRACT: Rat mast cell granules were obtained by homogenization of highly purified rat mast cells and isolated in a Percoll gradient. Diphosphoinositide (DPI) synthesis in rat mast cell granules was assayed by measuring the incorporation of 32P from [gamma 32P]ATP into DPI in the absence of exogenous phosphatidylinositol (PI). Lipids were isolated with methanol/chloroform/HCl and were separated by thin-layer chromatography on oxalic acid impregnated silica gel plates. DPI areas were identified by staining with iodine, scraped and measured for 32P radioactivity. The addition of phorbol myristate acetate (PMA) to the granules caused an increase of 32P incorporation from [gamma 32P]ATP in the DPI fraction, which can be catalyzed by PI kinase. This effect of PMA in the DPI synthesis was dose dependent and maximal effects were observed at 10 ng/ml.
International archives of allergy and applied immunology 02/1989; 90(4):373-7.
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Nihon Naika Gakkai Zasshi 01/1989; 77(12):1859-62.
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Arerugī = [Allergy] 01/1989; 37(12):1183-9.
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Nihon Kyōbu Shikkan Gakkai zasshi 12/1988; 26(11):1224-8.
