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Biomedical subjects

Y Okayama

Publications and source records attributed to Y Okayama.

At least 55 records · Page 3Linked to original sources
PubMed1991

Biological properties of human skin mast cells.

Mast cells and basophils, although sharing many constitutive properties, are quite distinct in their development, functions and biological properties. Mast cell granules are composed of a macromolecular matrix of proteoglycan and neutral protease of which heparin and tryptase, respectively, are predominant. The distribution of the other major neutral protease, chymase, allows human mast cell subpopulations to be subdivided immunocytochemically. All human mast cells respond to IgE-dependent stimulation with the secretion of the preformed mediator, histamine, and the newly generated lipid-derived eicosanoids PGD2 and LTC4. Although amounts of these products vary between mast cells dispersed from different tissues, it is uncertain whether this reflects true heterogeneity. Mast cells of the human skin, but not those of other tissues, are sensitive to stimulation by substance P, compound 48/80 and other basic non-immunological stimuli. The mechanism of mediator secretion induced by these agents is distinct from that induced by IgE-dependent stimulation. However, the morphological characteristics of degranulation are similar, suggesting that the distinct biochemical pathways merge into a common pathway before effecting degranulation.

Humans↗
PubMed1991

Mediator secretion from human skin mast cells provoked by immunological and non-immunological stimulation.

Mast cells of the human skin not only release mediators following immunological activation, but may also be stimulated to release histamine by the neuropeptides substance P, vasoactive intestinal polypeptide and somatostatin or by other basic secretagogues such as morphine, poly-L-lysine and compound 48/80. Release of histamine under these conditions is rapid and accompanied by minimal generation of the eicosanoids, prostaglandin (PG)D2 and leukotriene (LT)C4. Transient elevations of intracellular calcium are associated with mediator secretion induced by both stimuli, that induced by anti-IgE being derived from extracellular sources through channels in the plasma membrane while that stimulated by neuropeptides is mobilized intracellularly. Similarly, elevations of intracellular cyclic adenosine monophosphate (AMP) induced by anti-IgE occur only in the presence of extracellular calcium whereas with substance P elevations are apparent even in the absence of extracellular calcium. With the latter stimulus, histamine release is complete before the peak cyclic AMP is achieved. Histamine release stimulated by both secretagogues is unaffected by sodium cromoglycate or nedocromil sodium but is reduced by both salbutamol and isobutylmethylxanthine. Despite these biochemical and temporal differences, degranulation induced by both secretagogues proceeds by compound exocytosis which is indistinguishable under the electron microscope.

Calcium↗
PubMed1990

Rat mast cell granules reacting with a mouse monoclonal antibody M6764 which recognizes the same epitope of a mouse monoclonal antibody HNK-1.

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.

Animals↗
PubMed1990

Nontoxic concentration of ceftazidime and flomoxef sodium for intravitreal use--evaluated by in-vitro ERG.

The effects of ceftazidime (CAZ) and flomoxef sodium (FMOX) on the in-vitro electroretinogram (ERG) of albino rabbits were studied. The a-wave, the b-wave and the oscillatory potential (OP) were unchanged by 0.3mM (0.19mg/ml) CAZ-containing solution. The OP was suppressed by 0.5mM (0.32mg/ml) CAZ. The a-wave, the b-wave and the OP were unchanged by 0.5mM (0.26mg/ml) FMOX. The OP was suppressed and its peak latency was delayed by 2mM (0.52mg/ml) FMOX. The concentration of 0.3mM (0.19mg/ml) CAZ was higher than its minimum inhibitory concentration (MIC) against Staphylococcus aureus, Staphylococcus epidermidis, Pseudomonas aeruginosa, Serratia marcescens and Propionibacterium acnes. The concentration of 0.5mM (0.26mg/ml) FMOX was higher than its MIC against Staphylococcus aureus, Staphylococcus epidermidis, Serratia marcescens and Propionibacterium acnes.

Animals↗
PubMed1989

A diphosphoinositide kinase in rat mast cell granules.

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.

Adenosine Triphosphate↗
PubMed1989

Inhibitory effect of adenine nucleotides and anti-allergic drugs on phosphorylation of phosphatidylinositol in rat mast cell granules.

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.

1-Phosphatidylinositol 4-Kinase↗
PubMed1989

Phorbol myristate acetate stimulates formation of diphosphoinositide in rat mast cell granules.

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.

Animals↗
PubMed1989

Effect of staurosporine on histamine release from rat serosal mast cells.

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.

Alkaloids↗
PubMed1989

[Studies on thermal stimulation accompanying visible-light irradiation].

Visible-light-cured composite resin is one of the most frequently used materials in clinical practice. The use of this material inevitably involves dental pulp stimulation, i.e., thermal stimulation due to visible-light irradiation, during the restoration procedure. To determine the variations of temperature involved in visible-light irradiation, the author examines differences in temperature generated by various types of visible-light curing units. Changes in the temperature under varying conditions of irradiation were also examined. The results were as follows: 1) Among the visible-light curing units used in the present study, the highest temperature was obtained from Translux CL (a rise of 17 degrees C) whereas Hiliomat showed the lowest value (a rise of 7 degrees C). 2) The temperature increased with prolongation of irradiation time, reaching a maximum level (a rise of 15 degrees C) at 60s. 3) The temperature increased with shortening of irradiation distance, reaching a maximum level (a rise of 20 degrees C) at 3mm.

Composite Resins↗
PubMed1989

[Effect of phorbol myristate acetate (PMA) on diphosphoinositide (DPI) synthesis in rat mast cell granules].

Rat mast cell granules were obtained by homogenization of highly purified rat mast cells and isolated in a Percoll gradient. 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/HC1 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 PMA to the granules caused an increase of DPI synthesis, which can be catalysed by PI kinase. Neither an inactive phorbol ester, 4-alpha-phorbol-12, 13-didecanoate, nor dimethyl sulfoxide (DMSO) used as a solvent for PMA had any effect. The effect of PMA in the DPI synthesis was dose-dependent and maximal effects were observed at 10-100 ng/ml. Dose-response curves of the effects of PMA in DPI synthesis in the granules corresponded to those of other biochemical effects of PMA in rat mast cells, such as mediator release mediated through the activation of protein kinase C. These results suggest that PMA may directly affect PI kinase or indirectly regulate its activity in rat mast cell granules.

1-Phosphatidylinositol 4-Kinase↗
PubMed1988

Methotrexate-induced resistance to dactinomycin in choriocarcinoma.

NaUCC-2, a choriocarcinoma cell line, was derived from a patient who had a very poor clinical response to combination chemotherapy. Methotrexate (MTX) might have inhibited the antitumor effect of dactinomycin. To investigate this point, in vitro studies were performed to determine the sensitivity and uptake of MTX and dactinomycin (administered individually and in combination) to NaUCC-2 and three other choriocarcinoma cell lines. Dihydrofolate reductase (DHFR) concentrations were studied as well. Although NaUCC-2 showed sensitivity to MTX and dactinomycin, which were comparable to the other cell lines when they were given separately, NaUCC-2 was unique in that the combination of MTX and dactinomycin was less lethal than dactinomycin given by itself. The uptake of MTX in NaUCC-2 was significantly higher than that in the other cell lines, and MTX also induced an increase in dactinomycin uptake in NaUCC-2. There was no significant difference in DHFR activity. Although additional studies are necessary to determine the mechanism responsible for this effect, these findings suggest that a mechanism other than drug uptake or DHFR activity must play a role in the drug resistance for choriocarcinoma. These findings also suggest that the most commonly used combination chemotherapy for choriocarcinoma, dactinomycin and MTX, may not always be the best method.

Adult↗
PubMed1988

Nontoxic concentration of amphotericin B for intravitreal use--evaluated by in vitro ERG.

Amphotericin B (AMPH)-induced changes of the electroretinogram (ERG) were studied in the in vitro eye-cup of the albino rabbit. The a-wave, b-wave, oscillatory potentials and c-wave were not changed by 1 microM AMPH, and these all were slightly suppressed by 5 microM AMPH. These changes were mostly reversible. The b-wave and oscillatory potentials were greatly suppressed by 50 microM AMPH. These changes were only partially reversible 75 minutes after re-perfusion without AMPH.

Amphotericin B↗
PubMed1988

Measurement of portal blood flow by an ultrasonic "duplex" system composed of a pulsed Doppler flowmeter and a linear-type real time B mode electroscanner.

The portal vein hemodynamics of patients with various liver diseases were investigated by means of a duplex system consisting of a linear electroscanner and a pulsed Doppler flowmeter. In cases of chronic liver disease, the cross-sectional area of the portal vein trunk became greater as the liver injury proceeded, while the maximum velocity tended to decrease. However, blood flow volume was no different between the controls and the chronic liver disease group. On the other hand, the splenic venous flow volume tended to increase as the liver injury advanced, suggesting that the increase in splenic venous flow volume is closely related to the formation of esophageal varices. This method permits non-invasive observation of changes during the course of various types of liver disease.

Blood Flow Velocity↗
PubMed1987

Effects of antibiotics on the in vitro ERG of the albino rabbit. Penicillins and cephalosporins antibiotics.

This study describes the effects of penicillin G (PC-G) potassium, PC-G sodium, cloxacillin sodium (MCIPC), disodium sulbenicillin (SBPC), cefazolin sodium (CEZ) and cefsulodin sodium (CFS) on the in-vitro electroretinogram (ERG) of the albino rabbit. The b-wave and oscillatory potentials (OPs) were unchanged by 0.1 mM PC-G potassium or PC-G sodium. The OPs were slightly suppressed by 0.3 mM of either drug. While the a- and b-waves were not deteriorated, the OPs were greatly suppressed by 1.0 mM concentration. The effect of PC-G on the ERG was characterized by a selective suppression of the OPs. The b-wave and OPs were not suppressed by 0.03 mM MCIPC. They were slightly suppressed by 0.05 mM MCIPC. The a-wave, b-wave and OPs were not deteriorated by 1.0 mM SBPC. The b-wave and OPs were suppressed by 3.0 mM or 6.0 mM SBPC respectively. These changes appeared to be dose-dependent. Since the b-wave and OPs were concomitantly suppressed by both MCIPC and SBPC, these antibiotics, unlike PC-G, did not selectively suppress the OPs. The b-wave and OPs were unchanged by 0.1 mM CEZ or CFS. The OPs were slightly suppressed by 0.3 mM CEZ or CFS. CEZ or CFS of 1.0 mM did not deteriorate the a- and b-waves, but selectively suppressed the OPs. The effects of CEZ and CFS on the ERG were characterized by a selective suppression of the OPs. The above-described changes in the ERG were reversible.

Animals↗
PubMed1987

[The study of drug sensitivity on newly established three choriocarcinoma cell lines].

After serial transplantation in nude mice, we had established three new human choriocarcinoma cell lines (NaUCC-1,2 and 3). These three cell lines and BeWo were examined for sensitivity to Act-D, MTX and the combined agents (Act-D + MTX) 3H-Act-D uptake and 3H-MTX uptake, and were compared for each treatment. NaUCC-1 showed low sensitivity to MTX (p less than 0.05), but showed high sensitivity to Act-D (p less than 0.05). BeWo showed low sensitivity to Act-D (p less than 0.05). In examining sensitivity to the combined agents (Act-D+ MTX), the sensitivity of NaUCC-2 to Act-D was decreased (p less than 0.05) by MTX added at the same time. In the 3H-Act-D uptake experiment, NaUCC-1 did uptake a relatively larger amount of 3H-Act-D (p less than 0.01) and BeWo a smaller amount of it (p less than 0.05). In the 3H-MTX uptake experiment, NaUCC-1 did uptake a relatively smaller amount of 3H-MTX (p less than 0.05), but NaUCC-2 uptook a larger amount of it (p less than 0.005). NaUCC-1 established from the patient in whom tumor cells were resistant to treatment, had a low response to MTX. NaUCC-2 was established from the patient in whom it was found that MTX inhibits the Act-D effect on tumor cells. In the study of combined agents in NaUCC-2, the growth inhibition effect of Act-D was suppressed by the MTX added.

Animals↗