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N Hashimoto

Publications and source records attributed to N Hashimoto.

At least 379 records · Page 21Linked to original sources
PubMed1994

Identification of protein-tyrosine phosphatases prevalent in adipocytes by molecular cloning.

Protein-tyrosine phosphatases (PTPases) are among the fastest growing family of enzymes that are closely linked to signal transduction pathways involving reversible tyrosine phosphorylation. In order to identify PTPase homologs expressed in adipocytes that might regulate the action of insulin or growth factors in this tissue, we screened a rat adipocyte cDNA library at reduced stringency with a panel of candidate PTPase probes. After subcloning and sequence analysis of the positive plaques, this approach enabled us to identify the expression of LRP/RPTP-alpha, PTPase 1B, SH-PTP2/Syp, and LAR in adipocytes at an abundance of 16, 7, 6 and 3 per million, respectively. Furthermore, a sequence variant of SH-PTP2/Syp was identified that may have significance in the tissue-specific activity of this enzyme. These data provide insight into PTPase homologs that may have a physiological role in the regulation of phosphotyrosyl turnover in hormone signalling pathways in adipocytes.

Adipocytes↗
PubMed1994

Parthenogenetic activation of oocytes in c-mos-deficient mice.

In Xenopus the c-mos proto-oncogene product (Mos) is essential for the initiation of oocyte maturation, for the progression from meiosis I to meiosis II and for the second meiotic metaphase arrest, acting as an essential component of the cytostatic factor CSF. Its function in mouse oocytes is unclear, however, as is the biological significance of c-mos mRNA expression in testes and several somatic tissues. We have generated c-mos-deficient mice by gene targeting in embryonic stem cells. These mice grew at the same rate as their wild-type counterparts and reproduction was normal in the males, but the fertility of the females was very low. The c-mos-deficient female mice developed ovarian teratomas at a high frequency. Oocytes from these females matured to the second meiotic metaphase both in vivo and in vitro, but were activated without fertilization. The results indicate that in mice Mos plays a role in the second meiotic metaphase arrest, but does not seem to be essential for the initiation of oocyte maturation, spermatogenesis or somatic cell cycle.

Animals↗
PubMed1994

Immunohistochemical study for basic fibroblast growth factor and fibroblast growth factor receptor I in pituitary adenomas.

Immunohistochemistry and RT-PCR of basic fibroblast growth factor (bFGF) and one of its receptors (FGFR-I) were performed in pituitary adenomas. Sixty percent of pituitary adenomas showed strong or moderate immunoreactivity to bFGF. The immunoreactivity for FGFR-I in tumor tissues showed positive correlation to that for bFGF (X2 = 6.176, P = 0.0456). Basic FGF-positive cells consisted of pituitary adenoma cells as well as folliculostellate cells and, their distribution was heterogeneous. Expressions of bFGF and FGFR-I were not related to cell proliferation of pituitary adenomas or hormones produced, suggesting that bFGF plays some role other than progression of pituitary adenomas.

Adenoma↗
PubMed1994

Role of polymorphonuclear leukocytes in galactosamine hepatitis: mechanism of adherence to hepatic endothelial cells.

To investigate the role of polymorphonuclear leukocytes in galactosamine-induced hepatic injury, we injected rats intraperitoneally with antiserum against rat polymorphonuclear leukocytes to deplete circulating neutrophils, then administered galactosamine plus lipopolysaccharide. Polymorphonuclear leukocytes in the hepatic sinusoids were increased after administration of galactosamine plus lipopolysaccharide, whereas pretreatment with the antiserum decreased the number of circulating leukocytes and reduced the mortality and the severity of hepatic injury. Serum collected 1 hr after galactosamine/lipopolysaccharide treatment enhanced in vitro polymorphonuclear leukocyte adherence to hepatic endothelial cells and induced leukocyte superoxide production. Intercellular adhesion molecule-1 expression on hepatic endothelial cells was also enhanced after stimulation with the serum. Polymorphonuclear leukocyte adhesion was partially inhibited by an antibody against tumor necrosis factor-alpha but not by superoxide dismutase. These results suggest that polymorphonuclear leukocytes play an important role in galactosamine-induced hepatic injury and that the accumulation and activation of leukocytes, as well as the enhanced expression of adhesion molecules on hepatic endothelial cells, can be induced by biologically active mediators such as tumor necrosis factor-alpha. In addition, prostaglandins E1 and E2 lessened the enhanced adherence of polymorphonuclear leukocytes and thus contributed to protection against hepatic injury.

Alprostadil↗
PubMed1994

Effect of nerve growth factor on delayed neuronal death after cerebral ischaemia.

We investigated the protective action of nerve growth factor (NGF) on delayed neuronal death, and we also studied the involvement of the 200 kDa neurofilament (NF200) cytoskeletal proteins. Wistar rats were divided into three groups: Group I, in which transient forebrain ischaemia was produced; Group II, ischaemic group which received intraventricular administration of artificial cerebrospinal fluid (CSF); and Group III, ischaemic group which received intraventricular administration of 2 micrograms of 2.5 S NGF. Forebrain ischaemia in these rats was produced by causing transient bilateral occlusion of the common carotid arteries and lowering the mean blood pressure to 50 mmHg for 8 minutes. On the 1st and 7th day after ischaemia we histologically examined neuronal death in the hippocampal CA 1 sector. On the 7th day after ischaemia, mean cell death (degenerative cell number/total cell number) was 87 +/- 9% in group I (n = 7), 51 +/- 36% in group II (n = 7), and 14 +/- 16% in group III (n = 8) (p < 0.05 vs. group II). The concentration of NF200 in the hippocampal homogenate was measured by the Western blotting method on the 1st and 7th day after ischaemia. On the 1st day it was found to be 67 +/- 11% of that in the control group in group I (n = 6), 73 +/- 21% in group II (n = 6), and 84 +/- 7% in group III (n = 6) (p < 0.05 vs. group II). The concentration of NF200 in all groups remained at the same level until the 7th day after ischaemia (each group, n = 6).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1994

Intra-arterial papaverine for the treatment of cerebral vasospasm after subarachnoid hemorrhage.

Intra-arterial infusion of papaverine was used clinically as a vasodilator for spastic arteries after subarachnoid hemorrhage, since in vitro experiment demonstrated that papaverine is one of the most potent vasodilators of human cerebral arteries following subarachnoid hemorrhage. The present report describes the successful treatment of cerebral vasospasm after subarachnoid hemorrhage by intra-arterial infusion of papaverine hydrochloride and discusses the experimental background of this treatment.

Cerebral Angiography↗
PubMed1994

Enhancement of infectivity of hantavirus in cell culture by centrifugation.

Centrifugation was introduced during virus adsorption to Vero E6 cells to improve the infectivity of hantavirus. Centrifugal adsorption of a stock solution of Hantaan virus strain 76-118 to a monolayer of Vero E6 cells enhanced virus infectivity depending on the centrifugation time and the centrifugal force. The maximum level of infectivity (3.1 x 10(6) FFU/ml) was enhanced after a 2 h centrifugation at 671 x g, which was almost 9-times higher than that of conventional adsorption of the virus at 37 degrees C for 1 h. Vero E6 cells were inoculated with a new hantavirus strain, KI-91-40, isolated with a low infectious titer (400 FFU/ml) from an urban rat and adsorbed by centrifugation. A higher virus titer was detected sooner compared to when using conventional adsorption. To analyze the mechanism of the enhancement, the centrifugation was carried out before and after virus adsorption. The infectivity was reduced when Vero E6 monolayers were centrifuged before virus inoculation. When the centrifugation proceeded after inoculation, the infectivity was almost equal to that without centrifugation. The infectivity was only enhanced when centrifugation was carried out during inoculation. These results indicate that centrifugation promotes a very early event of infection, probably attachment of the virus to cells.

Animals↗
PubMed1994

Kinetics and cross-reactivity of the virus-specific antibody-forming cells in mice during primary and secondary infection with Japanese encephalitis virus and related flaviviruses.

A modified antibody-forming cell assay was used to enumerate splenocytes secreting antibodies to Japanese encephalitis (JE) virus and four other flaviviruses in infected cells as the target antigens. The optimal viral antigen expression for the assay was standardized in the infected cells for each virus and the incubation time varied depending of the cell line and the virus. The kinetics of response of JE virus-infected mice readily showed IgG isotype switching. Antibody-forming splenocytes of mice primed or boosted with one flavivirus could distinguish, in variable proportions, the homologous virus antigen from heterologous ones depending on the serocomplex of the virus. Antibody-forming cells from flavivirus-infected mice were flavivirus specific as evidenced by the lack of recognition of Getah virus (alphavirus) antigen. After JE virus-infected mice were cross-primed with another flavivirus, the IgG-forming cell response resembled the secondary response to both viruses but had higher affinity to JE virus than to the cross-priming virus.

Animals↗
PubMed1994

Comparison of nucleotide sequences of M genome segments among Seoul virus strains isolated from eastern Asia.

The nucleotide sequences of the M genome segments of three Seoul virus strains (KI strains) which were isolated from urban rats inhabiting the same enzootic focus between 1983 and 1988 were compared. The viral cDNAs were amplified by PCR and were directly sequenced. The nucleotide sequences of KI strains were extremely homologous regardless of isolation year (less than 10 substitutions in 3651 nucleotides, less than 4 substitutions in 1133 amino acids). In addition, the nucleotide sequence of the KI strain isolated in 1983 (KI-83-262) was also quite similar to that of other Seoul viruses, which were isolated from laboratory rats in Japan (strain SR-11, 98.1% and B-1 strain, 96.5%), from an urban rat in Korea (Seoul 80-39, 96.5%) and from an urban rat in China (R22 strain, 93.4%). All possible N-glycosylation sites in the deduced amino acid sequences were conserved among all Seoul viruses examined. The nucleotide and amino acid sequences of Seoul virus strains were highly conserved although they were isolated from various districts of eastern Asia. These results indicate the genetic stability of Seoul virus strains maintained under a natural environment and the homology of Seoul viruses isolated from various districts of eastern Asia. The relationship among Seoul virus strains isolated from eastern Asia was compared by phylogenetic analysis.

Amino Acid Sequence↗
PubMed1994

Dopamine has inhibitory and accelerating effects on ischemia-induced neuronal cell damage in the rat striatum.

Dopaminergic (DAergic) influence on ischemic neuronal cell damage in the dorsolateral striatum was studied. Intact and 6-hydroxydopamine (6-OHDA) lesioned rats, with and without pretreatment by D1 and D2 DA antagonists, were subjected to 20 min forebrain ischemia. Extracellular DA and glutamate (Glu) were measured using microdialysis technique. Histological examination was performed on the dorsolateral striatum and the hippocampal CA1 area 24 h after ischemia. DA increased 400-500 times the control level during ischemia among the groups except the 6-OHDA lesioned group. No significant changes were observed in the concentration of 3,4-dihydroxyphenylacetic acid (DOPAC), but a transient decrease was seen in homovanillic acid (HVA). Due to ischemia, Glu increased up to about 5 times the control level among the groups. Neuronal damage in the dorsolateral striatum was slightly attenuated by 6-OHDA lesion. Treatment by spiperone (D2 antagonist, 7 micrograms/kg IP) alone attenuated the damage strongly. Treatment by SCH23390 (D1 antagonist, 2.5 mg/kg IP) alone or both D1 and D2 antagonists had no effects. Data suggest that excessive Glu and DA are involved in neuronal cell damage. DA might enhance the damage via D2 but inhibit via D1 receptor.

Animals↗
PubMed1994

Renin inhibitor: transport mechanism in rat small intestinal brush-border membrane vesicles.

The transport characteristics of the renin inhibitor ((3S,4S)-4-[N-morpholinoacetyl-(1-naphthyl)-L-alanyl-N-methyl-(4-t hiazolyl)-L- alanyl]amino-3-hydroxy-5-cyclohexyl-1-(4-pyridyl)-1-pentanone; CH3-18) in rat small intestinal brush-border membrane vesicles (BBMV) were examined by a rapid filtration technique. The uptake of CH3-18 was markedly stimulated by an inwardly directed H+ gradient (pH 7.5 inside, pH 5.5 outside) and showed an uphill transport. It was competitively inhibited by tripeptides and tetrapeptides, but not by amino acids or dipeptides. A countertransport effect on the uptake of CH3-18 was observed in the vesicle preloaded with a tripeptide. Effects of the fragments of several renin inhibitors were evaluated by their inhibitory and countertransport effects on the uptake of CH3-18. The morpholino group at the N-terminal was found to be important for the uptake of CH3-18.

Amino Acid Sequence↗
PubMed1994

Renin inhibitor: relationship between molecular structure and oral absorption.

Common problems in developing renin inhibitors are low solubility, insufficient oral absorption, and fast hepatic clearance. We focused on the molecular structure of renin inhibitors to overcome these problems. Cyclodextrins (CD) improved the low solubility of renin inhibitors, with beta-CD showing the best ability to dissolve renin inhibitors. The intestinal absorption of renin inhibitors varied with both their solubility and molecular structure. Coadministration of beta-CD improved the intestinal absorption of some renin inhibitors with low solubility as measured by transport into the mesenteric vein in the absorption experiment using the rat intestinal loop. Substitutions at both the N and C terminals was essential for absorption from the small intestine. A naphthyl group at the N-terminal further improved intestinal absorption. A carrier system appeared to be involved in the intestinal absorption of some renin inhibitors. N-methylation at the amide bond of thiazolylalanine suppressed the high hepatic clearance of one of the test compounds 18 which was well absorbed from the small intestine and it improved its oral bioavailability.

Animals↗
PubMed1994

Ischemia induces the expression of the platelet-derived growth factor-B chain in neurons and brain macrophages in vivo.

To elucidate the role of the platelet-derived growth factor (PDGF)-B chain in the brain, we examined its expression in rat brains with focal ischemia. Focal ischemia was induced by permanent tandem occlusion of the middle cerebral and common carotid arteries in spontaneously hypertensive rats (SHRs). Northern analysis demonstrated that ischemia transiently increased mRNA expression of the PDGF-B chain, but not the PDGF-A chain, in the injured neocortex. The larger transcript (3.5 kb) of the B chain gradually increased to threefold by 16 h, whereas the smaller transcript (2.6 kb) of the B chain markedly increased sixfold by 4 h. Immunohistochemistry revealed enhanced immunoreactivity in the neurons in the infarct and in the periinfarct area from 16 h to days 4-7, with a peak at 24 h. Furthermore, the brain macrophages that accumulated in the infarct showed intense immunostaining in their perinuclear region from days 2 to 14, with a peak at days 5-6. The present study demonstrates that ischemia induces the expression of the PDGF-B chain, first in neurons and later in brain macrophages, and suggests an important role of the PDGF-B chain in the healing process of the injured brain.

Animals↗
PubMed1994

Regulation of insulin receptor, insulin receptor substrate-1 and phosphatidylinositol 3-kinase in 3T3-F442A adipocytes. Effects of differentiation, insulin, and dexamethasone.

Insulin rapidly stimulates tyrosine kinase activity of its receptor resulting in phosphorylation of its cytosolic substrate insulin receptor substrate 1 (IRS-1), which in turn associates with and activates the enzyme phosphatidylinositol 3-kinase (PI 3-kinase). In the present study we have examined these three initial steps in insulin action during the differentiation of 3T3-F442A adipocytes and after treatment with dexamethasone or insulin. The differentiation of 3T3-F442A cells was characterized by a 13-fold increase in insulin receptor protein, a 9-fold increase in IRS-1, and a 10- and 4.5-fold increase in their insulin-stimulated phosphorylation, respectively. The mRNA expression of these two proteins showed a similar 8-fold increase during differentiation. In addition there was a 3.5-fold increase in PI 3-kinase protein [85 kilodalton (kDa) subunit] and a 16-fold increase in IRS-1-associated PI 3-kinase activity between day 0 and day 8 of differentiation. Dexamethasone (1 microM) treatment of differentiated cells induced a further 48% (P < 0.05) increase in insulin receptor level, but the autophosphorylation of the receptor was decreased by 31 +/- 1% (P < 0.02). At the same time there was a decrease by 56 +/- 4% (P < 0.005) in IRS-1 protein and by 31 +/- 1% (P < 0.001) in IRS-1 phosphorylation. The expression of insulin receptor mRNA was unchanged, but the expression of IRS-1 mRNA was decreased by approximately 75% after dexamethasone. By contrast, dexamethasone induced a 69% increase in the level of PI 3-kinase as determined by immunoblotting. The combined effect of decreased IRS-1 phosphorylation and increased PI 3-kinase protein was a minimal change (15% decrease) in the association/activation between IRS-1 and PI 3-kinase. Chronic treatment with 100 nM insulin induced a time- and dose-dependent decrease in insulin receptor and IRS-1 protein levels reaching a nadir of 34 +/- 5% (P < 0.005) and 39 +/- 5% (P < 0.01) of control levels after 24 h, respectively. There was an even more marked decrease in the phosphorylation level of these proteins. Chronic insulin treatment also produced a 30% decrease in PI 3-kinase protein levels and a approximately 50% decrease in the association/activation between IRS-1/PI 3-kinase. The expression of insulin receptor and IRS-1 mRNA was unchanged during chronic insulin treatment.(ABSTRACT TRUNCATED AT 400 WORDS)

3T3 Cells↗
PubMed1994

Epizootiological studies of hantavirus infection among urban rats in Hokkaido, Japan: evidences for the persistent infection from the sero-epizootiological surveys and antigenic characterizations of hantavirus isolates.

Epizootiological studies of hantavirus infection among urban rats were carried out through the surveys repeated 11 times at the same dumping ground area in 1983 to 1988. A total of 279 rats (Rattus norvegicus) were captured during the surveys. Sero-positive animals to hantavirus strain SR-11 were detected in all the surveys. Overall positive rate of rats 6 months old or more (94/128, 73.4%) was significantly higher than that of younger rats (23/151, 15.2%, x2 = 96.4, P < 0.001). Therefore, age dependent acquisition of hantavirus infection among rats was confirmed. Seven hantavirus strains, KI-83-262 (August, in 1983, designated as strain KI-262 in our previous report (2)), KI-85-1 and 85-2 (July in 1985), KI-88-4, 88-11, 88-15 and 88-24 (October, 1988) were isolated from lung tissues of adult rats which have high titers of neutralizing antibody. Although the serum specimens of virus carrier rats neutralized the infectivity of all the KI isolates, no apparent antigenic change in the isolates was detected by indirect immunofluorescent antibody (IFA) assay using polyclonal and monoclonal antibodies (MAbs) regardless of isolation years. However, neutralization test showed slight difference of antigenicity among KI strains. These results epizootiologically confirmed that hantavirus infected persistently among urban rats in a presence of neutralizing antibody.

Animals↗
PubMed1994

Induction of lung tumors in C3H strain mice after single or fractionated irradiation with X-rays.

Murine model for lung tumor induction was studied in C3H/He male mice, a strain with low spontaneous incidence of lung tumors. Dose-response relationships in lung tumor induction were compared following irradiation with single doses and split doses of X-rays to the thorax either at night or in the daytime. The tumor incidence after a single 1.25 Gy dose at night during the period of nocturnal activity almost reached the maximum level after a 5 Gy dose in the daytime. Proliferative activity determined by observing the labeling index with tritiated thymidine in the normal lung was low as a whole, but tended to decrease in the daytime. When the proliferative response was induced by X-irradiation, significantly higher activity was observed at night. These circadian fluctuations were thought to affect radiosensitivity and lung tumor induction in mice. When split doses or fractionated doses of X-rays were applied to the thorax, lung tumor incidence definitely increased. The incidence after two 7.5 Gy doses with a 12 hr-interval was 41%, 3-fold higher than that after a single 15 Gy dose. Moreover, fractionated whole body irradiations (three times at 3 Gy with 3-month-interval) after a single 7.5 Gy thoracic irradiation was most effective in increasing not only the incidence (47%) but also the multiplicity of the lung tumor. More than 30% of tumor-bearing mice had two or more tumors following thoracic and whole body irradiations, while only 10% of tumor-bearers had multiple tumors after single or fractionated thoracic irradiation alone.

Animals↗
PubMed1994

Lymphocytic hypophysitis presenting with diabetes insipidus: case report and literature review.

Lymphocytic adenohypophysitis is an autoimmune disorder of the anterior pituitary gland which usually occurs in a women in the postpartum period. It has been considered that lymphocytic hypophysitis is confined to the adenohypophysis sparing the neurohypophysis, and that diabetes insipidus is not a clinical feature of the disorder. Here we report the case of a 50-year-old woman with lymphocytic hypophysitis which presented with diabetes insipidus. MRI indicated homogeneous swelling of the whole pituitary gland, loss of the normal high intensity of the posterior pituitary, and thickening of the pituitary stalk. A biopsied specimen of the pituitary revealed diffuse lymphocytic infiltration. The diabetes insipidus was controlled by the administration of DDAVP. The anterior pituitary function was not greatly damaged, and no hormonal replacement therapy was necessary. We suggest that this case represents a variant of lymphocytic adenohypophysitis and/or lymphocytic infundibuloneurohypophysitis, in which the chronic inflammatory process involves the infundibulum, adenohypophysis and neurohypophysis.

Autoimmune Diseases↗