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Biomedical subjects

H Kamata

Publications and source records attributed to H Kamata.

At least 37 records · Page 2Linked to original sources

Overexpression of the alanine carrier protein gene from thermophilic bacterium PS3 in Escherichia coli.

The alanine transporter (alanine carrier protein, ACP) gene of thermophilic bacterium PS3 was previously cloned and expressed in a functionally active form in Escherichia coli cells. To achieve controlled overproduction of the ACP protein, we designed a plasmid encoding a fusion protein comprising ACP joined to the carboxyl terminus of the maltose binding protein (MBP-ACP). Upon transduction of the plasmid into E. coli RM1 cells defective in alanine/glycine transport, the transport activity was expressed even before induction with 1-thio-beta-D-galacto-pyranoside (IPTG), and increased slightly on induction with IPTG at low concentrations. However, overexpression of the MBP-ACP gene, induced by higher concentrations of IPTG, resulted in death of the host cells. Hence we screened other host cells and found that the MBP-ACP fusion protein was produced in a large quantity in E. coli TB1 cells 3 h after IPTG induction. The MBP-ACP fusion protein was accumulated in cytoplasmic membranes in an amount reaching more than 20% of the total membrane protein. The affinity-purified MBP-ACP exhibited very low transport activity when reconstituted into proteoliposomes.

Amino Acid Sequence↗

Biological roles of angiotensin II via its type 2 receptor during rat follicle atresia.

Type 1 angiotensin II (ANG II) receptors play crucial roles in the regulation of blood pressure and fluid osmolarity, whereas the physiological roles of type 2 ANG II receptors (AT2) remain unclear. Because AT2 is expressed in atretic follicles where granulosa cells undergo apoptosis, we examined the space and time relationship between AT2 expression and follicle atresia in vivo and the effect of AT2 on follicle-stimulating hormone (FSH) actions in vitro. Binding studies, autoradiography, and RT-PCR of AT2 revealed that the AT2 content in granulosa cells was time dependently increased at both protein and mRNA levels in equine chorionic gonadotropin-treated immature female rats. This increase paralleled the progression of atresia. ANG II suppressed FSH-caused prevention of DNA fragmentation, increases in luteinizing hormone receptor content, and estrogen production through AT2 in cultured granulosa cells. Moreover, FSH-induced stimulation of extracellular signal-regulated kinase activity, critical for cell survival, was inhibited by AT2 stimulation. These results suggest that AT2 mediates the progression of follicle atresia through granulosa cell apoptosis by inhibiting FSH actions.

Angiotensin II↗

Immunizing effect of vaccinia virus expressing the nucleoprotein of rinderpest virus on systemic rinderpest virus infection in rabbits.

A recombinant vaccinia virus (RVV) expressing the nucleoprotein (NP) of rinderpest virus (RPV) was examined in rabbits for the involvement of the NP protein in protection from the RPV infection. Despite their production of anti-NP antibody, the RVV-immunized rabbits succumbed to the RPV challenge, although there was a slight delay in the onset of disease after the low-dose challenge. On the other hand, the animals immunized with RVV expressing the hemagglutinin (H) protein of the RPV were completely protected. These results indicate that the NP protein might be not so effective as the H protein for the protection against viremic and systemic infection with RPV.

Animals↗

Distribution of G serotypes and P genotypes of bovine group A rotavirus isolated in Japan.

OBJECTIVE: To identify the distribution of G serotype and P genotype of bovine group A rotavirus in Japan. DESIGN: Detection, isolation, electropherotyping, G serotyping and P genotyping of bovine group A rotavirus in 167 diarrhoeal faecal samples. PROCEDURE: Bovine group A rotavirus was detected and isolated, and bovine group A rotavirus isolates were identified electropherotype by polyacrylamide gel electrophoresis and G serotype and P genotype by reverse transcription-poly-merase chain reaction. RESULTS: Twenty-eight bovine group A rotavirus strains were isolated from 167 samples. Electropherotypes of the bovine group A rotavirus isolates were identified as long-genome electropherotype and the most prevalent combination of G serotype and P genotype of the bovine group A rotavirus isolates was G6P5 (25/28; 89.3%), followed by G6P11 (2/28; 7.1%) and G10P11 (1/28; 3.6%). CONCLUSION: These results suggest that the bovine group A rotavirus exhibiting G6P5 is the most common in Japan.

Animals↗

Seroprevalence against Toxoplasma gondii in domiciled cats in Japan.

A serological survey with latex agglutination test to detect anti-Toxoplasma gondii antibodies was conducted on 800 serum samples collected from domiciled cats at animal hospitals in various areas of Japan. The overall prevalence was 6.0% (48/800). Among 48 positive individuals, there was no specific distribution of strength of antibody titers; the titers were 1:64 in 8 cats, 1:128 in 12, 1:256 in 8, 1:512 in 10, 1:1,024 in 8 and 1:2,048 in 2. The maximum prevalence was 15.4% in 13 cats at 17-23 yrs old group, whereas all were negative in 58 cats aged 12-16 yrs. The age groups in the order of higher prevalence were 8, 4, 10, 5, 3, and 7 yrs, showing no aging effect to the prevalence. In terms of rearing conditions of those cats, they were classified into 4 groups, i.e., indoor, free, outdoor, and others. The prevalence in the outdoor group (11.1%) was significantly higher (p < 0.05) than that in the free group (4.8%). Epidemiological aspects observed in the domiciled cats were different from those reported in the stray cats.

Animals↗

Influences of alkaline ionized water on milk yield, body weight of offspring and perinatal dam in rats.

The authors previously reported that male offspring of mothers rats given alkaline ionized water (AKW) showed a significantly higher body weight by day 14 after birth than did offspring of mother rats given tap water (TPW); furthermore, marked myocardial necrosis and fibrosis were observed particularly in the former male offspring at the age of 15 weeks. In the present experiment we looked for differences in bioparameters, namely the milk yield of mothers and suckled milk volume of the offspring, between the AKW- and the TPW-treated groups in order to reveal the factors which cause the unusual body weight gain in the offspring. Even though we were able to repeat our previous observation (the body weight of the male offspring of the AKW group increased significantly more by day 14 and 20 after birth and of the female by day 20 after birth than did that of the TPW group (p < 0.05), no significant difference was noted in any of the bioparameters, including those related to milk production and consumption. It is thus suspected that the water-hydrated cation, which was transferred either to the fetus through the placenta or to the offspring through the milk, might be the cause of the unusual body weight increase. Since calcium plays an important role in skeletal formation, it is tentatively concluded that the higher calcium concentration of AKW enriched the mother, serum calcium which was transferred to the fetus through the placenta and to the offspring through the milk.

Alkalies↗

Histopathological influence of alkaline ionized water on myocardial muscle of mother rats.

We have reported that a marked necrosis and subsequent fibrosis of myocardium occurred among male rats 15 weeks old given alkaline ionized water (AKW) during gestation and suckling periods, and after weaning. In this study, it was examined whether similar lesions would occur in mother rats which were given AKW from day zero of gestation to day 20 of lactation. The myocardial lesion in the mother rats given AKW showed cell infiltration, vacuolation and fibrosis in the papillary muscle of the left ventricle, as were observed in male rats of 15 weeks old. Myocardial degeneration may cause a leakage of potassium into the blood that results in a higher concentration of potassium in the blood in the test group than in that of the control group given tap water.

Alkalies↗

Clinical features of ovarian cancer in Japanese women with germ-line mutations of BRCA1.

We analyzed the clinical features of 25 ovarian cancer patients who were associated with germ-line mutations of BRCA1 from four site-specific ovarian cancer families and seven breast-ovarian cancer families in Japan. The average age at diagnosis was 51.1 years (range, 38-77 years). Histological examination revealed 24 serous cyst adenocarcinomas in 25 patients. In 23 patients with clear clinical records, 3 patients had stage I disease, 17 had stage III disease, and 3 had stage IV disease. Thirteen patients with stage III disease who were treated with cisplatin-containing chemotherapy following tumor reduction surgery showed more favorable outcomes in both the survival rate and disease-free intervals, compared with age- and treatment course-matched controls (5-year survival rate, 0.786 versus 0.303; median disease-free interval, 91.43 versus 40.92 months; P < 0.05 for both, by logarithmic rank test). Our statistical model for the inheritance of susceptibility to ovarian cancer was derived from the analysis of 26 patients and 19 healthy carriers of 12 families. The expected lifetime risk of ovarian cancer is about 80% for women with mutations of BRCA1. These results suggest that the clinical outcome of ovarian cancer with germ-line mutations of BRCA1 appears to be more favorable than that with sporadic cases and that the disease penetrance among pedigrees with germ-line mutations of the BRCA1 gene is substantially high.

Adult↗

Differential effects of chronic administration of the antidepressants amitriptyline and rolipram on adenylyl cyclase activity.

Significant increases were observed in guanylylimidodiphosphate- and fluoride-stimulated adenylyl cyclase activities in synaptic membrane preparations from rat cerebral cortex subsequent to the repeated administration of rolipram and amitriptyline. The potency of GppNHp-dependent inhibition of adenylyl cyclase was decreased by chronic treatment with rolipram but not by amitriptyline treatment. These findings suggest that chronically administered rolipram and amitriptyline share the augmentation of adenylyl cyclase activity by different actions on post-receptor signaling.

Adenylyl Cyclases↗

Experimental infection in SPF kittens with a particular calicivirus strain originally isolated from lion.

Two SPF kittens were experimentally infected via the intranasal route with a strain of calicivirus originally isolated from sick lion suffering from vesicular disease. Fever, 40.3 degrees C and 40.7 degrees C, and vesicular formation in tongue and snout were reproduced in both kittens. The infected virus was recovered from nasal, oral and rectal swabs. A longer duration of virus recovery was proved with oral swab samples taken from 1 to 10 and 12 days post infection. This suggests that tongue and oral tissues are the main tissues for virus multiplication.

Administration, Intranasal↗

A possible immunosuppressant, cycloprodigiosin hydrochloride, obtained from Pseudoalteromonas denitrificans.

Cycloprodigiosin hydrochloride (cPrG.HCl), a member of the prodigiosin family, is a red pigment obtained from the marine bacterium Pseudoalteromonas denitrificans. cPrG.HCl markedly suppressed 3H-thymidine incorporation by concanavalin A stimulated murine splenocytes but had little effect on lipopolysaccharide dependent 3H-thymidine incorporation, indicating that cPrG.HCl acts as a selective inhibitor of T cell proliferation in the same way as other members of the prodigiosin family. cPrG.HCl inhibited the proliferation of the PMA stimulated Jurkat cells through an apoptotic process. Intriguingly, cPrG.HCl inhibited the H+ translocation by vacuolar type ATPase in chromaffin granule membranes without any effect on either its ATPase activity nor on the membrane conductance of phospholipid bilayers, suggesting that cPrG.HCl selectively uncouples H+ translocation from the ATPase reaction rather than acting as a non-specific ionophore. Since crystalline cPrG.HCl is highly stable, it raises the possibility of its therapeutic use as an immunosuppressant.

Animals↗

Redox regulation of lipopolysaccharide (LPS)-induced interleukin-8 (IL-8) gene expression mediated by NF kappa B and AP-1 in human astrocytoma U373 cells.

LPS-induced expression of the IL-8 gene was markedly enhanced by H2O2 or by deprivation of the cellular antioxidant glutathione by L-buthionine-(S,R)-sulfoximine (BSO) in human astrocytoma U373 cells. In contrast, it was markedly suppressed by the reductant N-acetyl-L-cysteine (NAC) and other antioxidants. Transient expression analysis using the chloramphenicol acetyltransferase assay revealed that activation of the IL-8 promoter by LPS was stimulated by BSO and was suppressed by NAC; likewise LPS-induced activation of both NF kappa B and AP-1 was enhanced by BSO and inhibited by NAC. These results suggest that LPS-induced IL-8 gene expression is regulated by cellular redox via modulation of these transcription factors.

Acetylcysteine↗

[Semi-supervised exercise using a step machine at home after myocardial infarction].

Home exercise programs for patients with myocardial infarction effectively improve their ability to exercise as well as quality of life. However, there are no efficient methods for monitoring the patient's clinical status or conveying the physician's instructions to the home setting. To resolve these problems, we developed a computer-based, automated, telemetry system comprised of central and peripheral computers and telephone line. Five myocardial infarction patients were evaluated for peak oxygen uptake (peak VO2) and anaerobic threshold (AT) before hospital discharge. At home, the patients performed 10-min exercise using a step machine. Patient data including blood pressure, pulse rate, and electrocardiogram before and after exercise were stored in the peripheral computer, and the central computer automatically retrieved the data through the phone line. If the current heart rate was less than the heart rate at AT, extreme changes in blood pressure were noted or dangerous arrhythmias appeared, appropriate instructions were indicated on the display of the peripheral computer. Following these instructions, the patients continued home exercise programs for 6 months. Peak VO2 and AT increased significantly in all patients (peak VO2: baseline 24 +/- 3.3 ml/kg/min, 6 months later 33.3 +/- 3.7 ml/kg/min, p < 0.01, AT: baseline 15.2 +/- 2.7 ml/kg/min, 6 months later 18.5 +/- 2.8 ml/kg/min, p < 0.01). The computer-based automated telemetry system combined with a step machine facilitated effective prescription and monitoring of exercise programs at home.

Adult↗

A strain of calicivirus isolated from lions with vesicular lesions on tongue and snout.

In December 1992, 17 African lions and 7 Siberian tigers in a Safari park in Japan became sick with characteristic clinical symptoms of acute vesicular formations on tongue and snout. The disease was highly contagious since all of these animals showed similar symptoms within two days after the onset of the first case. Swabs were taken from affected animals in rubbing tongues, snouts and some from rectums. Cytopathic viruses were isolated on CRFK cell culture by virological tests. The physicochemical property of a representative virus strain, named Arthur/L, isolated from a male lion was identified as a member of Caliciviridae. However, seroneutralization test indicated that this virus strain was antigenically distinct from Japanese isolates of feline caliciviruses used for comparison. Viral capsid proteins of the present isolate, Arthur/L, and of a feline calicivirus, strain FC7, were compared in an electrophoresis in SDS-PAGE gel. The major viral capsid polypeptide of them were proved to be significantly different in molecular weight. The polypeptide of FC7 was estimated to be ca. 63 KDa whereas that of Arthur/L consisted of 2 components of ca. 65 and 62 KDa. The viral proteins of these two strains were also proved to be distinct by an immunoblotting test.

Animals↗

Viral susceptibility of a newly established cell line derived from the peritoneal cavity of BALB/C mouse.

Viral susceptibility of a newly established cell line, named KMP, derived from the peritoneal cavity of BALB/C mouse is described. The cells were originally cloned from the in vitro culture of ascites of the mouse injected with Ehrlich ascitic tumor cells in advance. The electrophoretic pattern of cellular DNAs, extracted from KMP, normal BALB/C mouse spleen, and Ehrlich tumor cells respectively were compared after triple digestions with restriction endonucleases. This cell line was proved to be of mouse origin, but not the sub-line of Ehrlich tumor cells. The strains of Coxsackie virus B group, swine enterovirus, influenza virus, encephalomyocarditis virus, vesicular stomatitis virus, and Aujeszky's disease virus were able to multiply well in the cell line with considerably high infectious titers in showing clear CPE and circular plaques.

Animals↗

Suppression of nerve growth factor-induced neuronal differentiation of PC12 cells. N-acetylcysteine uncouples the signal transduction from ras to the mitogen-activated protein kinase cascade.

The cellular redox state is thought to play an important role in a wide variety cellular signaling pathways. Here, we investigated the involvement of redox regulation in the nerve growth factor (NGF) signaling pathway and neuronal differentiation in PC12 cells. N-acetyl-L-cysteine (NAC), which acts as a reductant in cells both by its direct reducing activity and by increasing the synthesis of the cellular antioxidant glutathione, inhibited neuronal differentiation induced by NGF or by the expression of oncogenic ras in PC12 cells. NAC suppressed NGF-induced c-fos gene expression and AP-1 activation. These results suggest that neuronal differentiation and NGF signaling are subject to regulation by the cellular redox state. NAC also suppressed the NGF-induced activation of mitogen-activated protein kinases (MAPKs) and decreased the amount of tyrosine phosphorylation of MAPKs. The suppression of MAPK by NAC was independent of glutathione synthesis. In parallel with the suppression of MAPK, the activation of MAPK kinase kinase activity was also suppressed in the presence of NAC. In contrast, NGF-induced activation of Ras was not inhibited by NAC. The inhibitory effect of NAC on the MAPK cascade was independent of transcription and translation. Thus, NAC suppresses NGF-induced neuronal differentiation by uncoupling the signal transduction from Ras to the MAP kinase cascade in PC12 cells.

Acetylcysteine↗

Nerve growth factor and forskolin prevent H2O2-induced apoptosis in PC12 cells by glutathione independent mechanism.

PC12 cells died by apoptosis at relatively low concentrations of H2O2, of which cytotoxicity was effectively suppressed by nerve growth factor (NGF), forskolin, and dbt-cAMP. Treatment with NGF or forskolin for 24 h increased the level of cellular antioxidant glutathione (GSH) by 1.6-2.0-fold. However, both NGF and forskolin protected cells against H2O2-stress even when cellular GSH was depleted by treatment with L-buthionine-(S,R)-sulfoximine (BSO). The GSH-independent protection effects of NGF and forskolin did not require new protein or RNA synthesis. Exogenous expression of an oncogenic ras suppressed apoptosis caused by H2O2 indicating that Ras protein also plays a role in suppressing apoptosis caused by oxidative radical stress.

Animals↗