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Biomedical subjects

H Ochiai

Publications and source records attributed to H Ochiai.

At least 109 records · Page 6Linked to original sources

[A case of mitochondrial cardiomyopathy with heart failure, sick sinus syndrome and diabetes mellitus: mitochondrial DNA adenine-to-guanine transition at 3243 of mitochondrial tRNA(LEU)(UUR) gene].

A 42-year-old woman with diabetes mellitus lost consciousness and was transferred to the Yokohama City University Hospital. Blood chemistry findings indicated low blood sugar levels and chest X-ray examination revealed cardiomegaly and bilateral pleural effusions. These clinical abnormalities were corrected by treatment with glucose, diuretics, angiotensin converting enzyme inhibitor and digitalis. Cardiological laboratory examinations were performed after admission. Electrocardiography revealed first degree atrioventricular block and incomplete right bundle branch block. Ultrasonography showed lower grade of ejection fraction and diffuse hypokinesis of the cardiac wall. After admission, sinus arrest suddenly occurred. The diagnosis was sick sinus syndrome. Scintigraphy using iodine-123 betamethyl-p-iodophenyl-pentadecanoic acid showed abnormal mottled defects. Coronary angiography found no significant stenosis of the coronary artery. Electron microscopy showed abnormally shaped mitochondrial accumulations in an endomyocardial biopsy specimen. Mitochondrial DNA amplification by polymerase chain reaction followed by restriction enzyme Apa I digestion revealed adenine-to-guanine transition at 3243 of the mitochondrial tRNA(LEU)(UUR) gene.

Adult↗

Nitric oxide and murine coxsackievirus B3 myocarditis: aggravation of myocarditis by inhibition of nitric oxide synthase.

OBJECTIVES: This study sought to investigate the effects of nitric oxide inhibition in a murine model of coxsackievirus B3 myocarditis. BACKGROUND: Little is known about the contribution of nitric oxide to the pathophysiology of myocarditis. METHODS: Antiviral activity was tested in vitro using nitric oxide inhibition by treatment with activated macrophages of NG-nitro-L-arginine methyl ester. In the in vivo experiments, NG-nitro-L-arginine methyl ester and NG-nitro-D-arginine methyl ester (both at 100 micrograms/ml) were administered to C3H/He mice early (days 0 to 14) and late (days 14 to 35) after infection with coxsackievirus B3. RESULTS: In the in vitro experiments with interferon-gamma- and lipopolysaccharide-induced activated murine macrophages, treatment with the nitric oxide synthase inhibitor NG-nitro-L-arginine methyl ester, but not its inactive enantiomer NG-nitro-D-arginine methyl ester, restored coxsackievirus B3 titers. In the in vivo experiments in the early treatment group, myocardial virus titers were higher in NG-nitro-L-arginine methyl ester-treated than infected untreated animals, and both inflammatory cell infiltration and necrosis were more severe. In the late treatment group, more severe necrosis and more dense myocardial and perivascular fibrosis were observed in NG-nitro-L-arginine methyl ester-treated than in infected untreated animals. NG-Nitro-D-arginine methyl ester administration was ineffective. CONCLUSIONS: Nitric oxide inhibition increases myocardial virus titers, resulting in the aggravation of cardiac pathology in the early stage of coxsackievirus B3 myocarditis. In the late stage, it induces more severe cardiomyopathic lesions. Nitric oxide plays a defensive role in the pathogenesis of coxsackievirus B3 myocarditis.

Animals↗

Differential changes in the concentrations of cytokine-induced neutrophil chemoattractant (CINC)-1 and CINC-2 in exudate during rat lipopolysaccharide-induced inflammation.

Recently we have purified four neutrophil chemotactic factors from conditioned medium of rat granulation tissue. Two chemokines besides cytokine-induced neutrophil chemoattractant-1 (CINC-1, formerly called CINC) and CNC-3/macrophage inflammatory protein-2 (MIP-2) were novel chemoattractants, designated as CINC-2 alpha and CINC-2 beta. In the present report, we developed an enzyme-linked immunosorbent assay (ELISA) specific for CINC-2. The biotin-streptavidin sandwich ELISA for CINC-2 beta detected CINC-2 alpha and CINC- 2 beta equally well between 1 ug/mI and 300 ng/ml, but did not show cross-reactivity with CINC-1 and CINC-3. The concentrations of CINC-1 and CINC-2 in the exudate during rat lipopolysaccharide (LPS)-induced inflammation were determined. The CINC-1 concentration in the exudate reached a maximum at 4 h after LPS injection, whereas the CINC-2 level steadily increased up to 8 h. The number of infiltrated cells in the exudate increased linearly until 6 h and gradually up to 8 h. Increase in the cell number was correlated with total concentrations of CINC-1 and CINC-2. The results suggest that CINC-2 as well as CINC-1 plays an important role in accumulation of neutrophils into the inflammatory lesion of LPS- induced inflammation in rats.

Animals↗

123I-MIBG myocardial imaging in hypertensive patients: abnormality progresses with left ventricular hypertrophy.

Twenty-seven patients with essential hypertension were prospectively studied with 123I-labeled metaiodobenzyl-guanidine (123I-MIBG) to assess the presence and location of impaired sympathetic innervation in hypertrophied myocardium. Thirteen patients had left ventricular hypertrophy on echocardiography, and 14 had normal echocardiograms. The wash out ratio of 123I-MIBG in these two groups did not differ significantly (35.3 +/- 6.1 and 35.4 +/- 5.1) but was higher than in control subjects (29.4 +/- 6.7). The delayed heart-to-mediastinum count ratio was lower in the patients with hypertrophy than in the patients without hypertrophy (1.93 +/- 0.28 and 2.22 +/- 0.21; p < 0.05) and the control subjects (1.93 +/- 0.28 and 2.33 +/- 0.25; p < 0.05). On SPECT imaging, abnormalities in segmental uptake were frequent at the posterior and postero-lateral wall in both groups, although the hypertrophic group had more significant impairment. Our results lead to the hypothesis that hypertension in more advanced stages may be associated not only with hypertrophic changes but also with more advanced regional impairment of cardiac sympathetic innervation.

3-Iodobenzylguanidine↗

Interleukin-6 induces proliferation of rat hepatocytes in vivo.

BACKGROUND/AIMS: The aim of this study was to assess the effect of interleukin-6 (IL-6) on the proliferation of hepatocytes and to study the interaction between IL-6 and hepatocyte growth factor (HGF) in vivo. METHODS: IL-6 was injected at a dose of 200 micrograms/mg subcutaneously into rats every day for 14 days. Liver and blood samples were obtained at 1, 3, 7 and 14 days during IL-6 administration. Hepatocyte proliferative activity of sera was measured using 3H-thymidine incorporation into cultured rat hepatocytes. To evaluate the proliferative activity of the hepatocytes in tissue sections, hepatic DNA content and immunostaining of the liver tissue sections for proliferating cell nuclear antigen (PCNA) were performed. Plasma HGF levels were measured using specific EIA. In addition, total RNA was extracted from the liver and expression of HGF mRNA was detected by RT-PCR. RESULTS: The DNA contents of liver taken from IL-6-treated rats were increased during IL-6 administration compared with untreated rats. Sera taken from IL-6-treated rats at various intervals during administration also significantly increased 3H-thymidine incorporation by cultured rat hepatocytes compared with sera from untreated rats, suppressing 3H-thymidine incorporation at day 1 and 3 by anti-HGF antibody. IL-6 itself did not increase 3H-thymidine incorporation. Increased expression of PCNA in these hepatocytes was noted from 1 day after IL-6 administration, and at 14 days, the number of PCNA-positive cells was sevenfold greater than in the livers of untreated rats. However, plasma HGF levels showed a peak at day 1, decreased gradually from day 3, and became undetectable by day 14. HGF mRNA expression in livers of IL-6-treated rats was suppressed from day 3 to day 14 of IL-6 administration. CONCLUSIONS: These data show that IL-6 induces an early phase of liver cell growth in vivo and suggest that an increase level of HGF mediates this effect.

Animals↗

Molecular cloning and characterization of the cDNA for discoidin II of Dictyostelium discoideum.

By using monoclonal antibodies directed against discoidin II, we have isolated cDNA clones from axenically grown Ax-2 cells. One cDNA clone (D2) contained a 1.2-kb insert encoding the entire discoidin II protein, which is composed of 257 amino acid residues and has a calculated molecular mass of 28,574. The amino acid sequences, determined by Edman degradation of six tryptic peptides of discoidin II, were identical to those deduced from the cDNA sequence. The protein bears no resemblance to any proteins in the data banks, except that its sequence is 49% identical with the amino acid sequence of discoidin I. Discoidin II shares with discoidin I both a carbohydrate-binding site and an Art-Gly-Asp (RGD) sequence, which has been found in fibronectin in mammalian cells. With the onset of aggregation (8 h of development), a 1.3-kb discoidin II mRNA begins to accumulate. A similar pattern of regulation occurs at the protein level.

Amino Acid Sequence↗

GABAergic synaptic innervation of catecholaminergic neurons in the area postrema of the rat.

Immunoelectron microscopy using the preembedding double immunostaining technique was applied to examine synaptic relationships between gamma-aminobutyric acid (GABA)-ergic neurons and catecholaminergic neurons in the area postrema of the rat. The catecholaminergic neuronal somata received only a few synapses from GABAergic axon terminals, whereas the catecholaminergic dendrites received many synapses from GABAergic axon terminals. Most of the synapses were symmetrical. GABAergic axon terminals could be found presynaptically to two catecholaminergic dendrites or both catecholaminergic and nonimmunoreactive dendrites. Conversely, a few catecholaminergic axon terminals were found presynaptically to GABAergic dendrites although such synapses were few. These findings suggest that in the area postrema, GABAergic neurons may modulate the functions of catecholaminergic neurons and may also be influenced by them.

Animals↗

Antisense RNA inactivation of gene expression of a cell-cell adhesion protein (gp64) in the cellular slime mold Polysphondylium pallidum.

The gp64 protein of Polysphondylium pallidum has been shown to mediate EDTA-stable cell-cell adhesion. To explore the functional role of gp64, we made an antisense RNA expression construct designed to prevent the gene expression of gp64; the construct was introduced into P. pallidum cells and the transformants were characterised. The antisense RNA-expressing clone L3mc2 which had just been harvested at the growth phase tended to re-form in aggregates smaller in size than did the parental cells in either the presence or absence of 10 mM EDTA. In contrast, 6.5-hour starved L3mc2 cells remained considerably dissociated from each other after 5 minutes gyrating, although aggregation gradually increased by 50% during a further 55 minutes gyrating in the presence of 10 mM EDTA. Correspondingly, L3mc2 lacked specifically the cell-cell adhesion protein, gp64. We therefore conclude that the gp64 protein is involved in forming the EDTA-resistant cell-cell contact. In spite of the absence of gp64, L3mc2 exhibited normal developmental processes, a fact which demonstrates that another cell-cell adhesion system exists in the development of Polysphondylium. This is the first report in which an antisense RNA technique was successfully applied to Polysphondylium.

Cell Adhesion Molecules↗

A case of idiopathic portal hypertension complicated with narrowing of intrahepatic bile ducts.

A 37-year-old woman who had portal hypertension followed by splenomegaly, developed collateral blood flow of spleno-renal shunt and paraesophageal veins, esophageal varices and further with narrowing of intrahepatic bile ducts shown by endoscopic retrograde cholangiography was described. Liver function was almost normal except the slight elevation of serum alkaline phosphatase and gamma-glutamyl transpeptidase levels. However, endoscopic retrograde cholangiography revealed the narrowing of the intrahepatic bile ducts. The histological examination of biopsied specimen showed no prominent change in portal tracts and bile ducts without cell infiltration or fibrosis in the portal area. This case will be considered as idiopathic portal hypertension complicated by narrowing of the intrahepatic bile ducts.

Adult↗

Effect of hyperthermia on proliferation and deviation of carcinoembryonic antigen and squamous cell carcinoma-related antigen of human esophageal carcinoma cells in culture.

As a basic study of hyperthermia on malignant tumors, we investigated the kinetics of proliferative activity and the values of tumor markers carcinoembryonic antigen (CEA) and squamous cell carcinoma-related antigen (SCC) in a human esophageal carcinoma cell line, SGF-4, following a change of culture temperature. The temperature range allowing cultured SGF-4 cells to proliferate was from 37 degrees C to 40 degrees C. In an experiment examining the recovery of proliferative activity, no proliferative activity was observed after the cultured cells were exposed to 42 degrees C for 72 hours. The values of CEA and SCC as tumor markers were found to be increased in association with the cell damage due to the change of temperature. These markers could thus be useful as indicators for evaluations of hyperthermia therapy effectiveness.

Aged↗

Na-dependent glutamate transport in high K and high glutathione (HK/HG) and high K and low glutathione (HK/LG) dog red blood cells.

Na-dependent glutamate (Glu) influxes in the red blood cells of normal low K (LK), high K and high glutathione (HK/HG), and high K and low glutathione (HK/LG) dogs were compared. The ranges of the influxes in LK, HK/HG and HK/LG cells were 1.0-63, 62-174 and 1.3-26 mumol/1 cells per h, respectively. Some LK and HK/LG dogs had red blood cells with extremely low Glu influxes. In cells with extremely low Glu influxes, however, there were clear Na-dependent Glu influxes. In LK, HK/HG and HK/LG cells, the Km of Na-dependent Glu influx with respect to the medium Glu concentration were 17, 20 and 19 mM, respectively, and the half-maximal activation (K1/2) with respect to medium Na concentration was 39, 40 and 42 microM, respectively. By the addition of harmaline, a hallucinogenic alkaloid, the Vmax in LK cells was not affected and the Km was increased, while the Vmax was decreased and the Km increased in HK/HG and HK/LG cells. The Ki value with harmaline by means of Dixon plot in LK cells was 5.2 mM, against 1.8 and 1.9 mM in HK/HG and HK/LG cells, respectively. These results suggest that the difference in the Na-dependent Glu influxes between 2 HK groups was due to the varying quantity, not the quality, of the transporter.

ATP-Binding Cassette Transporters↗

Inhibitory effect of bafilomycin A1, a specific inhibitor of vacuolar-type proton pump, on the growth of influenza A and B viruses in MDCK cells.

We studied the effect of bafilomycin A1 (Baf-A1), a novel and highly specific inhibitor for vacuolar-type proton (V-H+) pump, on the growth of influenza A and B viruses in Madin-Darby canine kidney cells. Vital fluorescence microscopic study showed that Baf-A1 induced the complete disappearance of acidified compartments such as endosomes and lysosomes both in infected and uninfected cells by the treatment with 0.1 microM inhibitor for 1 h at 37 degrees C. In addition, virus growth was inhibited when Baf-A1 was present from 1 h before infection to the end of incubation, or added within as early as 5-10 min after infection. Conversely, the virus growth was recovered in correlation with the reappearance of acidified compartments after removal of Baf-A1. These data suggest that Baf-A1-sensitive V-H+ pumps are solely responsible for the acidification of endosomes and lysosomes, and thus Baf-A1 inhibits the growth of influenza A and B viruses by affecting the acidified compartments in which low pH is essential for the uncoating process of influenza virus growth at an early stage of infection.

Animals↗

The reciprocal synaptic relations between enkephalinergic neurons and catecholaminergic neurons in the area postrema.

A preembedding double immunostaining technique using antibodies against methionine-enkephalin and tyrosine hydroxylase was used to study synaptic relations between enkephalinergic and catecholaminergic neurons in the area postrema of the rat at the electron microscopic level. The large nuclei-containing cell bodies of the catecholaminergic neurons displayed well-developed Golgi apparatus. The catecholaminergic somata and dendrites received synapses from enkephalinergic axon terminals, and most of the synapses were symmetrical. Occasionally, the catecholaminergic axon terminals were also found to be presynaptic to the enkephalinergic dendrites. Because the enkephalinergic neurons have been reported to be involved in cardiovascular function and the catecholaminergic neurons involved in the vomiting behavior, the synapses observed in this study may provide morphological evidence of the relationship between the vomiting and cardiovascular functions that are triggered in the area postrema.

Animals↗

Markedly elevated levels of beta2-microglobulin in urine with measles viruria in patients with measles.

BACKGROUND: Measles is a highly contagious infectious disease in children. However, the pathogenesis has not yet been fully defined since susceptible cells to measles virus (MV) had not been developed. Recently, B95a cells, which are very susceptible to MV, have been reported. OBJECTIVES: To evaluate measles viruria in patients with measles, isolation of MV from urine was performed using B95a cells. STUDY DESIGN: Isolation of MV from supernates or sediments of urine in patients with measles was performed using B95a cells. The levels of beta2-microglobulin (MG) in urine and serum were also measured. RESULTS: MV was isolated from either supernates or sediments of urine. MV was isolated from 10 of 11 samples (91%) within 2 days of the appearance of a rash. MV was isolated from supernates of urine for up to 4 days after the appearance of a rash, and from sediments for up to 5 days. The levels of urinary beta2-MG were elevated within 2 days of the appearance of a rash. The levels of urinary beta2-MG with measles viruria were significantly higher compared to those without measles viruria. CONCLUSIONS: Measles viruria may occur early in all patients with measles and elevated levels of urinary beta2-MG in patients with measles may be the consequence of tubular injury.

Journal Article↗

Inhibitory effect of ferulic acid and isoferulic acid on murine interleukin-8 production in response to influenza virus infections in vitro and in vivo.

We investigated the effect of ferulic acid (FA) and isoferulic acid (IFA), which are active components of the rhizoma of Cimicifuga species used frequently as anti-inflammatory drugs in Japanese Oriental medicines, on murine interleukin-8 (IL-8) production in response to influenza virus infections in vitro and in vivo by antibody-sandwich enzyme-linked immunosorbent assay. In the in vitro study, the murine macrophage cell line RAW 264.7 was infected with influenza virus at a dose of 10 plaque forming units (PFU)/cell and cultured in the presence or absence of drugs. Both FA and IFA reduced the IL-8 levels in the 20-h conditioned medium in comparison with control in a dose-dependent manner. The effect of IFA was greater than that of FA: IL-8 levels were reduced to 43% and 56% of the control in the presence of 100 micrograms/ml of IFA and FA, respectively. In the in vivo study, mice were infected with 1,000 PFU of virus and received daily oral administrations of Cimicifuga heracleifolia extract (5 mg/mouse/day), FA (0.5 mg/mouse/day), IFA (0.125 mg/mouse/day), or phosphate buffered saline. The three drugs showed a tendency to reduce IL-8 levels in bronchoalveolar lavage (BAL) obtained 2 days after infection. Moreover, both FA and IFA also significantly reduced the number of exuded neutrophils into BAL. However, the drug administrations did not affect the virus yields in BAL. These data suggest that FA and IFA are novel and potent inhibitors of murine IL-8 production and might act as one of the main components of anti-inflammatory rhizoma of Cimicifuga species.

Animals↗