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Biomedical subjects

N Hiyama

Publications and source records attributed to N Hiyama.

10 recordsLinked to original sources
PubMed1985

Immunological studies of herpetic keratitis: induction of cytotoxic T lymphocytes in mouse splenocytes.

C3H/He and BALB/c mice were inoculated with herpes simplex virus (HSV) by corneal or intraperitoneal route. The mouse splenocytes were analyzed to determine the level and mode of cytotoxic T lymphocyte (CTL) induction by a cytotoxicity test, using HSV-infected L929 cells and 3T3 cells as the target cells. To enhance the activities of CTL, mouse splenocytes were restimulated by lipopolysaccharide-induced lymphoblasts (LPS-blasts) infected with HSV before assay. HSV infection by both corneal and intraperitoneal routes induced H-2 restricted T lymphocyte responses specific for HSV. This CTL response became detectable on day 6 in mice with herpetic keratitis and on day 4 with intraperitoneal infection, but 10 days after infection the CTL activities of both groups reached the same level. A high response level persisted as long as 28 days. These results suggest that the protective mechanism in herpetic keratitis is similar to the mechanism in intraperitoneal infection.

Animals↗
PubMed1983

Use of the immunoperoxidase technique for estimation of susceptibility of herpes simplex virus to IDU.

We applied the direct immunoperoxidase method to estimate the susceptibility to IDU of 17 strains of the virus which were isolated from the patients with active epithelial lesions of herpetic keratitis. All the strains were found to be sensitive below the IDU concentration of 5 micrograms/ml. The findings obtained by this method were in good agreement with those by the fluorescent antibody technique. The present method is specific and simple. It offers clear-cut interpretation and a permanent record of the results.

Drug Resistance, Microbial↗
PubMed1983

Effects of 5-iodo-2'-deoxyuridine on ultrastructural localization of viral antigens in cultured cells infected with herpes simplex virus.

Changes in viral morphology and localization of viral antigens induced by 5-iodo-2'-deoxyuridine (IDU) were studied using herpes simplex virus-infected monolayers of cultured rabbit corneal cells. A direct antibody enzyme method was employed for immunoelectron and light microscopy. There was no significant difference between the control and the IDU groups in the intracellular distribution of antigen up to 2 hours after infection. At 2 hours after infection, intracytoplasmic antigen began to appear in control and IDU-treated cells either in patchy distributions or in association with rough endoplasmic reticulum. Intranuclear staining appeared also at 2 hours after infection as a diffuse fine granular reaction. At 3 hours after infection, the intracytoplasmic reaction became more or less diffuse and there was slightly more cytoplasmic staining in the untreated group. At 6 hours after infection, reaction products increased in the cytoplasm and the nuclei in both groups but IDU-treated cells showed less intracytoplasmic and more intranuclear staining. The nuclear envelope was stained positive in both groups but IDU-treated cells showed a decreased reaction in the cytoplasmic membranous structure and fewer reaction deposits on the cell surface. At 12 and 18 hours after infection, the intranuclear accumulation of reaction products after IDU treatment became more evident. Progeny virus particles were significantly fewer in the presence of IDU: Nucleocapsids were found but in a greatly reduced number and only one incompletely enveloped particle was found. The morphology of the individual nucleocapsid, however, was not significantly altered by treatment with IDU. The preponderance of reaction substances in the nuclei compared to the cytoplasm in treatment groups may reflect distorted protein synthesis and may provide morphological evidence of the biochemical hypothesis of IDU efficacy: IDU exerts its antiviral effects through abnormal protein synthesis and the resultant defects in capsid assembly. Decreased reaction substances in the membranous structures of the cytoplasm may be related to reduced or abnormal production of envelope-associated viral antigens.

Animals↗
PubMed1980

Enzymic determination of urinary free L-fucose.

Conditions necessary for the precise measurement of free L-fucose in urine by an enzymic method using L-fucose dehydrogenase have been studied. The normal urinary levels of L-fucose were 16.4 /+- 9.1 micrograms/ml in children (22.2 /+- 6.5 micrograms/mg of creatinine) and 17.7 /+- 8.5 micrograms/ml in adults (16.6 /+- 5.7 micrograms/mg of creatinine). There was a close correlation between the concentration of free L-fucose and that of creatinine. A close correlation was also found between the concentration of free L-fucose and alpha-L-fucosidase activity. Thus, it was suggested that the free L-fucose in urine reflected the metabolism of L-fucose or L-fucose-containing glycoconjugates. In a preliminary screening test, several urine samples which showed high concentrations of free L-fucose, namely cases of "fucosuria" were found.

Adult↗
PubMed1979

Isolation and characterization of L-fucose dehydrogenase from rabbit liver.

L-Fucose dehydrogenase [EC 1.1.1.122] was isolated from a rabbit liver extract and purified about 390-fold with a yield of approximately 13%. The purification procedures included treatment with protamine, ammonium sulfate fractionation, treatment with acid, DE-32 celluose colum chromatography, gel filtration on Sephadex G-100, preparative polyacrylamide gel electrophoresis, and affinity chromatography on 5' AMP-Sepharose 4B. The last procedure, affinity chromatography on 5' AMP-Sephadex 4B, was useful for the removal of other dehydrogenases. The eznyme which was homogeneous, as shown by polyacrylamide gel electrophoresis, had a molecular weight of about 92,000. The optimum pH was at 10.0 and isoelectric point at 5.2. The enzyme accepted both L-fucose and D-arabinose as substrate, but was specific for NAD+ as coenzyme. Km values were 0.15 mM, 1.4 mM, and 0.7 mM for L-fucose, D-arabinose, and NAD+, respectively. A single enzyme catalyzed the oxidation of L-fucose and D-arabinose, which had the same configurations of hydroxyl groups from C-2 to C-4. The reaction products obtained with L-fucose as substrate were L-fucono-lactone and L-fuconic acid. The L-fucono-lactone was an immediate product of oxidation and was hydrolyzed to L-fuconic acid spontaneously. This reaction was irreversible. Therefore, it is likely that L-fucose dehydrogenase is involved in the initial step of the catabolic pathway of L-fucose in rabbit liver.

Animals↗