Norio Hayashi MD

Publications (3)34.99 Total impact

• Article: Hepatitis C viral quasispecies in hepatitis C virus carriers with normal liver enzymes and patients with type C chronic liver disease

  Masafumi Naito, Norio Hayashi MD, Toyoki Moribe, Hideki Hagiwara, Eiji Mita, Yoshiyuki Kanazawa, Akinori Kasahara, Hideyuki Fusamoto, Takenobu Kamada
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  ABSTRACT: Hepatitis C virus (HCV) has been reported to conform to a quasispecies nature, which is most evident in hypervariable regions of the putative envelope 2 domain. The aim of this study was to determine the relationship between the nucleotide complexity and diversity of hypervariable region 1 and various stages of the carrier states. The subjects studied were 20 HCV carriers with normal alanine aminotransferase (ALT) levels, 50 patients with chronic hepatitis who showed elevated ALT levels, 22 with cirrhosis, and 24 with hepatocellular carcinoma. The quasispecies complexity was analyzed by means of polymerase chain reaction-mediated single strand conformation polymorphism (PCR-SSCP). The value of nucleotide diversity was calculated by PCR cloning and sequencing. The number of SSCP bands ranged from 1 to 7, with no significant differences in the mean numbers among the stages of HCV infection. There was no correlation between the amounts of serum HCV RNA and the numbers of SSCP bands. No significant difference was found in the values of nucleotide diversity between carriers with normal ALT levels (mean, 6.6 × 10−2 per site) and patients with chronic hepatitis (7.7 × 10 −2). These findings suggest that the quasispecies complexity of hypervariable region 1 is independent of the stage of chronic HCV infection. (Hepatology 1995; 22:407–412.)
  Hepatology 12/2005; 22(2):407 - 412. · 11.66 Impact Factor
• Article: In vivo transfection of hepatitis C virus complementary DNA into rodent liver by asialoglycoprotein receptor mediated gene delivery

  Masato Yamamoto, Norio Hayashi MD, Yasuhide Miyamoto, Tetsuo Takehara, Eiji Mita, Makoto Seki, Hideyuki Fusamoto, Takenobu Kamada
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  ABSTRACT: An in vivo model of hepatitis C virus (HCV) infection is needed to enable investigation of the mechanism of the liver injury that it causes. In this study, we used asialoglycoprotein receptor mediated gene delivery to obtain expression of the complementary DNA (cDNA) coding the core and part of the envelope 1 protein of HCV because selective delivery to the hepatocytes has been reported to be attained with this method. The optimum carrier-DNA ratio was examined using in vitro transfection and found to be important for the efficiency of this method. In transfection in vivo, microautoradio-graphical examination showed that the transfected plasmids were delivered selectively to the liver parenchymal cells. To obtain an immunohistochemically detectable level of protein expression in rodent liver, some modifications for increasing the in vivo transfection efficiency were performed; a lysosomal enzyme inhibitor, chloroquine, was used and the administration route of the carrier-DNA complex was changed from the tail vein to the portal vein. On the bases of these results, in vivo transfection with expression vector of HCV core/E1 region was performed. In rat liver transfected by intraportal injection with chloroquine, the transcript RNA and the core protein were detected. These results indicated that the HCV core/E1 expression vector was not merely delivered but also successfully expressed in the liver using asialoglycoprotein receptor mediated gene delivery. The number of the HCV core expressing cells in the transfected liver was similar to that in patients with hepatitis C. These in vivo transfected animals should be useful for investigating the role of this region in the liver injury caused by HCV. (Hepatology 1995; 22:847–855.)
  Hepatology 08/1995; 22(3):847 - 855. · 11.66 Impact Factor
• Article: The significance of immunoglobulin M antibody response to hepatitis C virus core protein in patients with chronic hepatitis C

  Nobukazu Yuki, Norio Hayashi MD, Kazuyoshi Ohkawa, Hideki Hagiwara, Masahide Oshita, Kazuhiro Katayama, Yutaka Sasaki, Akinori Kasahara, Hideyuki Fusamoto, Takenobu Kamada
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  ABSTRACT: The significance of immunoglobulin (Ig) M antibody to hepatitis C virus core protein (IgM anti-HCVcore) was studied in 41 patients with chronic hepatitis C virus (HCV) infection diagnosed by the polymerase chain reaction (PCR). IgM anti-HCVcore was tested with a solid-phase enzyme-linked immunoassay. The results were correlated with clinical features, liver histology findings evaluated by the histological activity index, and virological features such as genotypes and viremic levels assessed by a branched DNA assay. IgM anti-HCVcore was found in 29 (71%) patients, and its occurrence was only related to viremic levels. A significant relationship was observed between viremic levels and IgM anti-HCVcore cut-off index (rs = .42, P < .01). Of the eight low viremic (branched DNA-negative) patients, only two (25%) tested positive for IgM anti-HCVcore with a low cut-off index of <3, whereas 27 (82%) of the 33 highly viremic (branched DNA-positive) patients had IgM anti-HCVcore (P < .01). After a 28-week interferon-α course (IFN-α), sustained aminotransferase normalization after therapy withdrawal was achieved by only two (13%) of the 16 patients with IgM anti-HCVcore cut-off index >3 compared with 11 (44%) of the 25 patients with that <3 (P < .05). IgM anti-HCVcore cut-off index decreased after therapy in patients who cleared the virus in sera but increased again after reappearance of viremia. These findings suggest that IgM anti-HCVcore may serve as a simple serological indicator of active virus replication and have relevance to the outcome of antiviral therapy. (Hepatology 1995; 22:402–406.)
  Hepatology 07/1995; 22(2):402 - 406. · 11.66 Impact Factor