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Biomedical subjects

M Satoh

Publications and source records attributed to M Satoh.

At least 829 records · Page 46Linked to original sources

Convulsants induce interleukin-1 beta messenger RNA in rat brain.

The effects of systemic administration of kainic acid and pentylenetetrazol on interleukin-1 beta gene expression in the rat brain was studied. After the administration of kainic acid in a convulsive dose (10 mg/kg i.p.), Interleukin-1 beta mRNA was induced intensely in the cerebral cortex, thalamus and hypothalamus, moderately in the hippocampus and weakly in the striatum, but not in the midbrain, pons-medulla and cerebellum. Pentylenetetrazol induced Interleukin-1 beta mRNA in the cerebral cortex, hypothalamus, and hippocampus with a faster time-course than kainic acid. Diazepam suppressed both the convulsion and the induction of Interleukin-1 beta mRNA produced by kainic acid. Dexamethasone suppressed the induction of Interleukin-1 beta mRNA, but did neither the convulsion nor the induction of c-fos mRNA following the injection of kainic acid. These results provide the first evidence that intensive neuronal excitation induces Interleukin-1 beta mRNA in particular regions of the brain.

Animals↗

Interspecies differences in the population of multiple glutamate receptors expressed in Xenopus oocytes after injection of guinea pig, mouse and rat forebrain mRNAs.

Expression of multiple glutamatergic responses was compared in Xenopus oocytes several days after injection of poly(A)+ RNAs from guinea pig, mouse and rat forebrains. No difference either in the characteristics of each response or in the amplitude of N-methyl-D-aspartate response was observed between the oocytes injected with mRNA from different species. However, the average magnitudes of kainate (KA) and oscillatory quisqualate (QA) responses produced by guinea pig brain mRNA were 1/2 and 1/6 times, respectively, of those by rat brain mRNA. These results may reflect that the guinea pig forebrain contains smaller amounts of ionotropic KA and metabotropic QA receptors than the rat forebrain.

Animals↗

Decreased proapolipoprotein A-I processing in liver disease: evidence for hepatic participation in proapolipoprotein A-I conversion.

We evaluated the proapolipoprotein A-I (proapo A-I)-converting activity to clarify the pathogenesis of the high proapo A-I/apo A-I ratio in the high-density lipoprotein (HDL) from patients with acute hepatitis and liver cirrhosis. The serum proapo A-I-converting activities were measured using 3H-labeled proapo A-I. 3H-labeled proapo A-I was purified from the media of cultured Hep G2 cells by immunoaffinity chromatography. The serum proapo A-I-converting activities were found to be significantly reduced in the patient with liver cirrhosis (140 +/- 53 dpm/ml per h) or acute hepatitis (140 +/- 48 dpm/ml per h) compared to normal subjects (315 +/- 32 dpm/ml per h). Serum proapo A-I-converting activity has a positive correlation with liver function tests such as serum albumin, choline esterase activity, ICG clearance and inverse correlation with proapo A-I/apo A-I ratio in HDL. These results suggest that the high proapo A-I/apo A-I ratio is due to the decreased proapo A-I-converting activity, and that the liver plays a significant role in the conversion of proapo A-I to apo A-I.

Amino Acid Sequence↗

The use of Xenopus oocytes to evaluate drugs affecting brain Ca2+ channels: effects of bifemelane and several nootropic agents.

Effects of nootropic or cerebroprotective drugs on voltage-sensitive Ca2+ channels (VSCC) in the mammalian brain were evaluated comparatively by depolarization-evoked Ca2+ channel currents in Xenopus oocytes injected with brain mRNA and by a high K(+)-stimulated 45Ca uptake into synaptosomes. It was found that several anti-amnesic agents (bifemelane, idebenone and vinpocetine) inhibited these VSCC-related responses with stronger potency than phenytoin and flurazepam. On the inhibiting potency of various compounds, there was a significant but weak correlation between the results from synaptosomes and those from the injected oocytes, since nifedipine, verapamil and diltiazem at 100 microM did not reduce 45Ca influx in synaptosomes but partly inhibited VSCCs in the oocytes. The blockade of neuronal L- and N-type VSCCs may participate in the anti-ischemic/hypoxic actions of nootropic drugs.

Animals↗

Evidence for receptor-mediated inhibition of intrinsic activity of GTP-binding protein, Gi1 and Gi2, but not G0 in reconstitution experiments.

The receptor-mediated inhibition of intrinsic activities of GTP-binding proteins (G-proteins) was studied. Pertussis toxin (IAP)-substrate G-protein, Gi1, Gi2 or G0, was prelabeled with [alpha-32P]GDP and reconstituted with synaptic membranes of the guinea pig cerebellum in the presence of 0.02% of Chaps. Intrinsic activities of G-proteins were evaluated by the release of [alpha-32P]GDP in exchange for added GppNHp or GDP in reconstituted preparations. U-50,488H (1 nM-10 microM), a specific kappa-subtype of opioid receptor agonist, inhibited the [alpha-32P]GDP release in exchange for added 1 microM GppNHp in Gi1-reconstituted preparations in a concentration-dependent manner. On the other hand, the kappa-opioid agonist at 10 microM increases the Km values of GppNHp, but not GDP in exchange for [alpha-32P]GDP release in preparations reconstituted with Gi1 or Gi2, but not with G0. These findings indicate that kappa-opioid receptor is coupled to inhibition of intrinsic activities of Gi1 and Gi2, but not G0, in guinea pig cerebellar membranes. In addition, it was revealed that the mode of action is mediated by a decrease in affinity of GTP (or its analog) for G proteins, but not by a change in affinity of GDP.

3,4-Dichloro-N-methyl-N-(2-(1-pyrrolidinyl)-cycloh↗

Changes in glycolipid expression in human testicular tumor.

Glycolipids were extracted from testicular tumor tissues of 13 patients, and their pattern of expression compared with that of normal testicular tissue. The most conspicuous and consistent change in the tumor extracts was marked accumulation of CTH (ceramide trihexoside). Structural analysis by enzyme cleavage showed that CTH which accumulated in the tumor tissue was Gb3 (Gal alpha 1-4Gal beta 1-4Glc beta 1-Cer). Immunohistochemistry using anti-Gb3 monoclonal antibody (MAb) (1A4) also indicated massive accumulation of Gb3 in the tumor tissue. Gb3 may be a new marker of testicular tumors, especially seminomas, for which useful markers are so far lacking.

Biomarkers, Tumor↗

Interleukin-1 beta inhibits long-term potentiation in the CA3 region of mouse hippocampal slices.

The effect of recombinant human interleukin 1-beta (IL-1 beta) on long-term potentiation (LTP) in the mossy fiber-CA3 pathway of mouse hippocampal slice preparations was investigated. IL-1 beta significantly inhibited LTP in concentrations as low as 2.9 pM (50 pg/ml). This effect of IL-1 beta was blocked by concurrent application of 100 nM Lys-D-Pro-Thr, a tripeptide analogue of IL-1 beta. This is the first evidence that IL-1 beta can regulate LTP of synaptic transmission in the hippocampus.

Animals↗

Phosphorylated mu-opioid receptor purified from rat brains lacks functional coupling with Gi1, a GTP-binding protein in reconstituted lipid vesicles.

The effects of phosphorylation of a mu-opioid receptor on signal transduction to G-protein were studied. The mu-opioid receptor purified from rat whole brains was reconstituted with purified Gi1 in phosphatidylcholine vesicles. DAGO, a mu-opioid agonist at 1 microM-1 mM increased GTPase activity by 10-110% of control, in a concentration-dependent manner. When the mu-opioid receptor was phosphorylated by cyclic AMP-dependent protein kinase prior to reconstitution with Gi1, the DAGO-stimulation was markedly reduced (20% increase at 1 mM DAGO).

Animals↗

Kappa-opioid agonist inhibits phospholipase C, possibly via an inhibition of G-protein activity.

In synaptic membranes of guinea pig cerebellum, in which preparations U-50, 488H, a kappa-opioid agonist inhibits G-protein activity, the same kappa-agonist at 100 nM inhibited the phospholipase C (PLC) activity stimulated by 100 microM GTP in the presence of 10 microM Ca2+. The present study provides a unique hypothetical mechanism that the receptor-mediated inhibition of PLC is mediated by inhibition of G-protein activity.

3,4-Dichloro-N-methyl-N-(2-(1-pyrrolidinyl)-cycloh↗

Purification of a novel type of calcium-activated neutral protease from rat brain. Possible involvement in production of the neuropeptide kyotorphin from calpastatin fragments.

We have found a novel type of Ca2(+)-activated neutral protease in rat brain cytosol which cleaves -Tyr-Arg-containing calpastatin fragments to release the neuropeptide kyotorphin. This enzyme was purified about 26,000-fold by column chromatography as follows: DE52 cellulose, Ultrogel AcA 44, thiopropyl-Sepharose 6B, second DE52 cellulose, Ultrogel AcA 34, and blue Sepharose CL-6B. The molecular mass of the enzyme was estimated to be 65-75 kDa by gel filtration. The purified enzyme gave a single band of 74 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Some properties of this enzyme were similar to those of the calpains, i.e. an absolute requirement for Ca2+, maximal activity at neutral pH, and inhibition by sulfhydryl reagents such as p-chloromercuriphenylsulfonic acid and N-ethylmaleimide. However, it differs from the calpains in that it possesses no caseinolytic activity, separates from the calpains on the first DE52 column, and is insensitive to leupeptin and E-64 (N-[N-(L-3-trans-carboxyoxrian-2-carbonyl)-L-leucyl]agmatine). Thus, the molecular mass, the substrate specificity, the chromatographic behavior, and the inhibitor spectrum all suggest that this enzyme is a novel type of Ca2(+)-activated neutral protease.

Amino Acid Sequence↗

Purification of an endothelin receptor from human placenta.

We have identified an endothelin (ET) binding protein on the membranes of human placenta and purified it to homogeneity. It is a polypeptide with an apparent Mol. Wt. of 40,000 and is a major protein to be labeled by cross-linking with either 125I-ET-1, -2, or -3. Binding studies with Scatchard analysis indicated the presence of a single class, high-affinity binding site with Kds of 57 pM, 480 pM and 40 nM for 125I-labeled ET-1, ET-2 and ET-3, respectively. These results suggest that the 40K protein is a major ET receptor in placenta and, most likely, can bind differentially to ET-1, ET-2 and ET-3.

Affinity Labels↗

Effect of a factor VIII concentrate on type IIB von Willebrand's disease-associated thrombocytopenia presenting during pregnancy in identical twin mothers.

Marked thrombocytopenia developed during pregnancy in both identical twins mothers who had systemic lupus erythematosus (SLE) and also type IIB von Willebrand's disease (vWD). The proband's platelet count decreased in the third trimester of pregnancy. Large-dose gamma-globulin and prednisolone treatments were performed because of the suspicion of immune thrombocytopenic reaction associated with SLE. These treatments were not effective. Her platelet count returned to the normal range immediately after delivery. Postpartum examinations revealed the decreased ristocetin cofactor activity and the deficiency of large von Willebrand factor (vWF) multimers in preserved plasma samples from the third trimester. These abnormal findings improved after delivery. Investigation of family members revealed that the proband had inherited type IIB vWD from her mother. The other twin, who was also under treatment for SLE, became pregnant about 1 year after delivery in the proband and followed almost the same course as that observed in the proband. As bleeding tendency was observed a few days before delivery, a factor VIII concentrate (Haemate P) was administered to compete with her variant vWF. This concentrate could prevent the further decrease in her platelet count, thereby correcting the hemorrhagic tendency. It seems evident that factor VII concentrate would be effective in treating thrombocytopenia associated with type IIB vWD.

Adult↗

Chinese hamster ovary cells continuously secrete a cysteine endopeptidase.

The protease activity in serum-free conditioned medium of chinese hamster ovary (CHO) cells was measured using peptidyl (or aminoacyl)-4-methylcoumaryl-7-amides (MCAs) as the substrates. Aminopeptidase increased in level as amounts of nonviable cells increased during cultivation in serum-free medium, indicating that the activity seems to be originated from intracellular proteases. The activity toward Boc-Leu-Arg-Arg-MCA, which was strongly inhibited by p-chloromercuribenzonate and N-ethylmaleimide, was the strongest among those toward peptidyl-MCAs in the conditioned medium within 48 h-cultivation in serum-free medium. In contrast to the case of aminopeptidase activity, the endopeptidase activity decreased in level after 48 h-cultivation although amounts of nonviable cells increases. Thus, CHO cells continuously secrete the cysteine proteases.

Aminopeptidases↗

Different drug-susceptibilities of long-term potentiation in three input systems to the CA3 region of the guinea pig hippocampus in vitro.

The susceptibilities to several drugs of long-term potentiations in the three input systems (mossy, commissural/associational and fimbrial fibres) to CA3 pyramidal neurones were investigated in hippocampal slices from the guinea pig. D-2-Amino-5-phosphonovalerate (D-APV), a selective antagonist at N-methyl-D-aspartate (NMDA) receptors, blocked the long-term potentiations in the commissural/associational fibre- and fimbrial fibre-CA3 systems, but did not significantly affect that in the mossy fibre-CA3 system. The latter was suppressed by kynurenate, a non-selective glutamate receptor antagonist. On the other hand, naloxone, an opioid antagonist, inhibited and bifemelane, which improves metabolism in brain and has an anti-amnesic action, augmented long-term potentiation in mossy fibre-CA3 system but did not influence those in commissural/associational fibre- and fimbrial fibre-CA3 systems. These findings suggest that the mechanisms, relevant to production of long-term potentiation in the mossy fibre-CA3 system, are different from those in the commissural/associational fibre- and fimbrial fibre-CA3 systems. N-Methyl-D-aspartate receptors are involved in the latter systems, while non-NMDA receptors for L-glutamate and opioid receptors are involved in the former. Further, the mossy fibre-CA3 system is more susceptible to a drug, having an anti-amnesic action, than are the other two systems.

2-Amino-5-phosphonovalerate↗

Effect of surfactant on bioelectric properties of canine tracheal epithelium.

The functional significance of pulmonary surfactant in the airways is not well known and the effects of surfactant on bioelectrical properties of airway epithelium have not been investigated. In the present study, we examined the effect of synthetic surfactant (TR-14) or calf lung surfactant extract (surfactant TA) on transepithelial potential difference (PD) and short circuit current (SCC) in canine trachea. The conductance (G) was calculated as the ratio of SCC per open-circuited PD. The posterior membrane without muscular layer from canine trachea was mounted in an Ussing-type chamber, bathed with Krebs-Ringer buffer solution at 37 degrees C and gassed with 95% O2-5% CO2, pH 7.4. Treatment with mucosal surfactant produced an increase in both PD and SCC in a dose-dependent fashion. PD and SCC reached 132% and 124% of before control at 0.25 mg/ml after the addition of each surfactant respectively, whereas G remained unchanged. Both ouabain and furosemide abolished surfactant-evoked increases in PD and SCC, whereas amiloride did not alter the surfactant-evoked increases. No significant differences in the effect on bioelectric parameters were observed between TR-14 and surfactant TA. These findings suggest that lung surfactant affects bioelectrical properties and changes ion transport (Cl- secretion) across airway epithelium, probably through the activity of ion pumps in the cellular membrane.

Animals↗