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T Ishimatsu

Publications and source records attributed to T Ishimatsu.

34 records · Page 2Linked to original sources

Possible binding sites for inositol 1,4,5-trisphosphate in macrophages.

We reported that an arylazide derivative of inositol 1,4,5-trisphosphate (InsP3) caused irreversible InsP3-induced Ca2+ release from saponin-permeabilized macrophages after photoirradiation. To determine the specific receptors for InsP3, presumably present on the endoplastic reticulum, we synthesized isotope-labeled arylazide derivatives of InsP3; InsP3 was coupled with p-azidobenzoyl [3H]beta-alanine (InsP3-[3H]AB beta A) or p-[125I]azidosalicyl beta-alanine (InsP3-[125I]AS beta A). We report here that three proteins may be associated with the Ca2+ releasing mechanism, in photoirradiated saponin-permeabilized macrophages and in the microsomal fraction.

Animals↗

Effect of guanosine triphosphate on the release and uptake of Ca2+ in saponin-permeabilized macrophages and the skeletal-muscle sarcoplasmic reticulum.

The effects of GTP, with or without polyethylene glycol (PEG), on the release and uptake of Ca2+ were examined by using saponin-treated macrophages and sarcoplasmic reticulum isolated from skeletal muscles. The application of GTP in concentrations in the range 0.1-10 microM induced a gradual, small but sustained release of Ca2+ from the saponin-treated macrophages. The addition of PEG to GTP markedly enhanced the GTP-mediated Ca2+ release. GTP at the same concentration ranges used for Ca2+ release decreased the amount of Ca2+ uptake, at a steady state, but stimulated the rate of Ca2+ accumulation in the presence of oxalate, the Ca2+-precipitating anion. The addition of PEG abolished the GTP-evoked stimulation of Ca2+ accumulation in the presence of oxalate. The stimulating effect on the rate of Ca2+ accumulation by GTP and its elimination by PEG were not due to changes in the permeability of oxalate by either GTP or PEG, or both. The Ca2+-releasing effect of GTP without PEG was enhanced by eliminating the uptake activity by decreasing the content of ATP. These results indicate that GTP has an inherent activity to release Ca2+ from non-mitochondrial intracellular stores of saponin-treated macrophages, and PEG enhances the GTP-mediated Ca2+ release, partly owing to its eliminating effect on GTP-stimulated Ca2+ uptake activity. These effects of GTP observed with saponin-permeabilized macrophages were not apparent in the isolated skeletal-muscle sarcoplasmic reticulum.

Animals↗

Localization of ferritin in human liver diseases studied by immuno-histochemical and immuno-electron microscopic procedures.

Localization of ferritin using a pre-embedding diffusion technique and an indirect localization sequence has been made in 34 cases of human liver under normal and several pathological conditions. With light microscopic observation, positive immuno-staining for ferritin was demonstrated as diffuse deposits in the hepatocytes and Kupffer cells. Intensity of the positive immuno-staining for ferritin in these cells appeared to roughly coincide with serum ferritin levels of each patient, but showed no disease specificity, although hepatoma cells contained weak deposits or were negative from immuno-staining for ferritin. With electron microscopic studies, intracellular antigen was well preserved in the hepatocytes and Kupffer cells in most cases with the positive immuno-staining for ferritin being observed in cytosol and a few cisternae of rough endoplasmic reticulum. Content of the positive immuno-staining for ferritin differed considerably from one case to another and one cell to another even in the same case. There was no immuno-staining for ferritin in hemosiderin pigment, lysosome, most of rough endoplasmic reticulum, Golgi complexes, and nucleus in both cells.

Ferritins↗

Mechanisms of flunarizine-induced vasodilation in the rabbit mesenteric artery.

The vasodilating effects of flunarizine on smooth muscle strips of rabbit mesenteric artery have been investigated and compared with those of nifedipine. Flunarizine (30-300 nM) dose-dependently inhibited Ca2+-induced contractions in Ca2+-free solution containing 100 mM K+. Double reciprocal analysis showed that this inhibition was either competitive at low concentrations (30-100 nM; nifedipine-like) or noncompetitive at high concentrations (0.3-1 microM). The latter seemed to be partly related to an inhibition of contractile proteins as estimated from Ca2+-induced contractions in saponin-treated chemically skinned muscle strips. In contrast to the actions of nifedipine, flunarizine inhibited norepinephrine (NE)-induced contractions more than those induced by high K+, and at 0.3 microM, this agent totally blocked NE-induced contraction. Flunarizine also inhibited NE-induced contraction in Ca2+-free solution containing 2 mM EGTA. In Ca2+-free solution, NE rapidly hydrolyzed phosphatidylinositol 4,5-bisphosphate (PI-P2) and produced phosphatidic acid (PA). Flunarizine (30 and 300 nM), but not nifedipine (100 nM), inhibited NE-induced hydrolysis of PI-P2 and production of PA. However, flunarizine (100 nM) did not modify the contraction induced by 10 microM inositol 1,4,5-trisphosphate in chemically skinned muscle strips. It is concluded that flunarizine inhibits both voltage-dependent (nifedipine-like) and receptor-operated Ca2+ influx induced by NE and also inhibits NE-induced Ca2+ release from intracellular stores due to inhibition of the hydrolysis of PI-P2.

Animals↗

Inflammatory activity of a trypsin-like protease in the tuberculin reaction sites in guinea pig skin.

A trypsin-like protease(s) was found in the extracts from tuberculin reaction sites in the guinea pig skin. Extractable enzyme activities were chronologically paralleled with the gross appearance of the inflammation, and the maximum activity from 24-hour old inflamed sites was about 12 times stronger than that from control skin, suggesting a potential role in the pathogenesis of the delayed hypersensitivity reaction (DHR). The enzyme suitably hydrolyzed t-butyloxycarbonyl-phenylalanyl-seryl-arginine 4-methylcoumaryl-7-amide, and was partially purified by gel filtration. It showed an apparent molecular weight of 700,000 with optimal pH of 9.0-9.5. The activity was inhibited by various serine protease inhibitors but not by thiol or metal protease inhibitors. It was relatively heat-stable. These findings suggest that the protease is similar to a trypsin-like protease from DHR to bovine gamma-globulin. Intradermal injection of the enzyme into normal guinea pigs induced a mixed neutrophil-mononuclear infiltrate. The reaction was reduced in intensity with the diisopropylfluorophosphate-inhibited enzyme preparation. The enzyme did not induce any increased vascular permeability. These findings suggest that the protease may be present generally in the DHR sites and probably participates in the cellular response of DHR.

Animals↗

Mechanism of vasodilation induced by alpha-human atrial natriuretic polypeptide in rabbit and guinea-pig renal arteries.

Effects of alpha-human atrial natriuretic polypeptide (alpha-HANP) on electrical and mechanical properties of smooth muscle cells of the guinea-pig and rabbit renal arteries and of the guinea-pig mesenteric artery were investigated. alpha-HANP (up to 10 nM) modified neither the membrane potential nor resistance of smooth muscle cells of the guinea-pig and rabbit renal arteries. In the guinea-pig mesenteric and renal arteries, alpha-HANP (up to 10 nM) had no effect on the amplitude and facilitation (mesenteric artery) or depression (renal artery) of excitatory junction potentials nor on action potentials. In the guinea-pig renal artery, alpha-HANP (up to 10 nM) had no effect on the depolarization induced by noradrenaline (NA) (up to 10 microM) but markedly inhibited NA-induced contraction. alpha-HANP (10 nM) slightly inhibited the K-induced contraction. In the rabbit renal artery, alpha-HANP (10 nM) inhibited the NA-induced contraction and to a lesser extent the K-induced contraction. In the rabbit renal artery, the effects of alpha-HANP on the release of Ca from the cellular storage by two applications of NA, and its re-storage, were investigated in Ca-free solution containing 2 mM-EGTA. When 5 nM-alpha-HANP was applied before and during the first application of 0.5 microM-NA, the contraction was markedly inhibited but the contraction to a second application of 10 microM-NA was potentiated. If the first dose of NA was 10 microM the effect was very small. Under the same experimental procedures, nitroglycerine (10 microM) showed almost the same effects as alpha-HANP on the NA-induced contractions. When both the first (3 mM) and second (10 mM) contractions were evoked by caffeine in Ca-free solution, alpha-HANP (5 nM) and nitroglycerine (10 microM) inhibited both contractions to the same extent. In the rabbit renal artery, applications of alpha-HANP or nitroglycerine increased the amount of guanosine 3',5'-phosphate (cyclic GMP) in a dose-dependent manner. However, a much higher concentration of nitroglycerine was required (2 X 10(3) times). In the rabbit renal artery, hydrolysis of phosphatidyl inositol 4,5-bisphosphate (PI-P2) activated by 0.5 microM-NA was inhibited by alpha-HANP, in a dose-dependent manner, but activation by 10 microM-NA was not inhibited by alpha-HANP (up to 100 nM).(ABSTRACT TRUNCATED AT 400 WORDS)

Action Potentials↗

[Clinical significance of CEA levels in lung cancer].

Serum concentrations of CEA, Ferritin, beta 2-MG, TPA and IAP were measured in patients with lung cancer. Significant difference of positive rate between lung cancer and benign chest disease was observed only in CEA and TPA. CEA elevations occurred most frequently with stage IV and histologically with adenocarcinoma. Positive CEA levels (greater than or equal to 5 ng/ml) of pleural effusion were observed in 75% of all patients with lung cancer and increased to 94% in patients with positive cytology of effusion. There was not significant difference of bronchial washings CEA levels between lung cancer and benign chest disease. Furthermore, serial measurement of postoperative CEA levels had considerable value in monitoring the course of disease following resection.

Adenocarcinoma↗

Assessment of the role of surgery for stage III bronchogenic carcinoma.

In order to assess the role of surgery for stage III bronchogenic carcinoma, we reviewed our results of 282 patients who were treated with surgical resection or various other therapy modalities. The survival in patients who underwent surgical resection was significantly better than those with nonresectable carcinoma. However, superior survival results were demonstrated only in patients with complete resection. In contrast, the survival in patients with palliative resection was similar to that in patients with exploratory thoracotomy and inoperable carcinoma. On the other hand, all patients with T3N2 lesions died of their disease within two years after surgery even when undergoing complete resection. Thus, we conclude that radical resection should be indicated in patients with T3N0-1 and T1-2N2 lesions if complete resection is expected.

Adenocarcinoma↗

A high-molecular-weight trypsinlike protease in the skin sites of delayed hypersensitivity in guinea pigs.

A trypsinlike protease was extracted from the delayed hypersensitivity skin sites in guinea pigs. Extractable amounts of the enzyme were chronologically paralleled with the gross appearance of the inflammation, and the maximum activity from the inflamed sites at 24-36 hours was about 20 times stronger than that from normal skin, suggesting a potential role in the pathogenesis of the delayed hypersensitivity reaction. The enzyme, which suitably hydrolyzed t-butyloxycarbonyl-phenylalanyl-seryl-arginine 4-methylcoumaryl-7-amide, was partially purified by isoelectric focusing or by gel filtration. The enzyme demonstrated a single peak of activity on the former column with an apparent isoelectric point of 4.2, and in the latter it showed an apparent molecular weight of 600,000 (600K-protease). When incubated with 3H-diisopropylfluorophosphate, the enzyme lost all amidolytic activity and yielded a single band of radioactivity in polyacrylamide disk gel electrophoresis in the presence of sodium dodecyl sulfate, and a single peak of radioactivity in gel filtration, both having an apparent molecular weight of 31,000-33,000 (31K-protease). That the 600K-protease might be a complex with alpha 2-macroglobulin was ruled out. The 31K-protease was separated from the 600K-protease by gel filtration in the presence of 6 M guanidine hydrochloride, and was renatured to an active form. An apparent isoelectric point of the 31K-protease observed was 9.4, suggesting that the 600K-protease may be a complex of 31K-protease with an acidic carrier molecule(s). Both proteases, 31K- and 600K-protease, had identical substrate specificity, a pH profile of amidolytic activity, and susceptibility to exogenous protease inhibitors. However, when sensitivities to intrinsic protease inhibitors in guinea pig plasma, two kinds of trypsin inhibitor, and alpha 2-macroglobulin were compared, the 600K-protease was at least 100 times more resistant than the 31K-protease. It was supposed that one of the pathophysiologically significant functions of the complex formation might be to maintain the enzyme activity longer in vivo.

Amidohydrolases↗

Guinea pig macroalbumin. A major inhibitor of activated Hageman factor in plasma with an alpha 2-macroglobulin-like nature.

A major inhibitor of the beta form of activated Hageman factor (beta-HFa) with an apparent molecular weight of 28,000, which was reported as a strong permeability factor (Yamamoto and Cochrane, Am J Pathol 1981, 105: 164-175), was purified from guinea pig plasma. When it was depleted in vitro, the plasma lost 71% of the total inhibitory activity toward beta-HFa. The inhibitor, termed "macroalbumin," with an apparent molecular weight of 720,000 and an apparent pI of 4.6, seemed to be an inhibitor similar to alpha 1- or alpha 2-macroglobulin of man and other mammalian species in physicochemical characteristics and in enzymologic properties. Though the inhibitory activity to beta-HFa was negligible in normal skin extract, a significant inhibitory activity appeared in extracts of permeability-enhanced skin sites which were induced by intradermal beta-HFa injection. The inhibitory activity that appeared was macroalbumin-dependent, with more than a 10-fold increase in the concentration. These results indicate the roles of macroalbumin as a negative feedback regulator in situ to the Hageman-factor-dependent pathway in a permeability enhancement system.

Animals↗

A case of amyloidosis associated with diphenylhydantoin therapy.

Prolonged administration of diphenylhydantoin (DPH) has been implicated as a possible etiologic factor in immunological aberrations and lymphoproliferative disorders. Diphenylhydantoin may account for the increase in susceptibility to lymphoproliferative diseases, as a result of its immunosuppressive effect. We report a case of amyloidosis with monoclonal gammopathy which developed during DPH treatment, without multiple myeloma or lymphoproliferative disorders. The association between DPH and monoclonal gammopathy is very rare, and such a case of amyloidosis associated with DPH has not been reported previously. DPH, however, may have played a role in the development of monoclonal gammopathy, which was the precursor of amyloid protein.

Aged↗

Relationship between nurse counseling and sexual adjustment after hysterectomy.

The relationship between nurse counseling of hysterectomy patients and sexual adjustment following surgery was the focus of this descriptive study. Of 108 premenopausal women admitted to seven hospitals in California, New Mexico, and Utah who completed a series of questionnaires preoperatively, postoperatively, and eight weeks post-surgery, only 11 percent identified the nurse as the person who supplied the most valuable information. Self-reported sexual adjustment following surgery was significantly (p=.001) related to their presurgical pattern. Almost half the respondents wrote comments urging nurses to initiate discussion about the effect of hysterectomy on sexuality.

Adaptation, Psychological↗