JaLCDOI 10.18926/AMO/32656
フルテキストURL fulltext.pdf
著者 Hada, Hajime| Koide, Norio| Hanafusa, Tadashi| Sakaguchi, Kosaku| Shinji, Toshiyuki| Sasaki, Shunsuke| Oka, Takahiko| Takayama, Niro| Yumoto, Yasuhiro| Tsuji, Takao|
抄録 <p>We detected an antibody to HCV envelope protein (E1) in sera of patients with HCV-related chronic liver diseases (20 patients with chronic hepatitis and 5 patients with liver cirrhosis) by Western blotting using the fusion protein of E1 envelope protein and beta-galactosidase as an antigen. The antibody to HCV E1 (anti-HCV E1) was detected in 8 (42%) of 19 patients positive for HCV-RNA (16 were positive and 3 were negative for antibody to C100-3) and in 1 (17%) of 6 patients negative for HCV-RNA but positive for antibody to C100-3. HCV-RNA was detected in 8 (89%) of 9 anti-HCV E1 positive sera. The value of alanine aminotransferase was significantly higher in patients positive for anti-HCV E1 than in patients negative for the antibody. Although an antibody to the envelope protein of HCV is suspected to be one of the candidates of virus-neutralizing antibodies, our results suggest this hypothesis appears to be unlikely.</p>
キーワード hepatitis C virus envelope antibody Western blotting
Amo Type Article
発行日 1992-10
出版物タイトル Acta Medica Okayama
46巻
5号
出版者 Okayama University Medical School
開始ページ 367
終了ページ 370
ISSN 0386-300X
NCID AA00508441
資料タイプ 学術雑誌論文
言語 English
論文のバージョン publisher
査読 有り
PubMed ID 1279946
Web of Sience KeyUT A1992JX49500007
JaLCDOI 10.18926/AMO/32196
フルテキストURL fulltext.pdf
著者 Hada, Hajime| Koide, Norio| Takabatake, Hiroyuki| Hanafusa, Tadashi| Tsuji, Takao|
抄録 It has been reported that the envelope region located at the 3' portion of the structural protein coding region is one of the most variable regions at both nucleotide and amino acid sequence levels in the hepatitis C virus (HCV) genome. We cloned HCV cDNA fragments of an envelope protein coding region (HCVNK), which were derived from serum of a Japanese patient with hepatocellular carcinoma and were amplified by polymerase chain reaction. After determining the nucleotide sequence, deduced amino acid sequence of the envelope protein region was compared with those of six HCV strains already published (HCJ1, HCVUS, HCJ4, HCVJH, HCVJ and HCVBK). Homology analysis among the strains revealed that the seven strains were classified into two subtypes; a US subtype (HCJ1 and HCVUS) and a Japanese subtype (HCJ4, HCVJH, HCVJ, HCVBK and HCVNK), since percentage homologies between two subtypes (70.3-77.3%) were significantly lower than those within each subtype (83.9-93.5%). Detailed analysis of the amino acid sequences also indicates that the region at aa246-aa258, tentatively named intersubtype variable region-1, may distinguish the US subtype from the Japanese subtype.
キーワード hapatitis C virus envelope DNA sequecing homology intersubtype variable region
Amo Type Article
発行日 1991-10
出版物タイトル Acta Medica Okayama
45巻
5号
出版者 Okayama University Medical School
開始ページ 347
終了ページ 355
ISSN 0386-300X
NCID AA00508441
資料タイプ 学術雑誌論文
言語 English
論文のバージョン publisher
査読 有り
PubMed ID 1661558
Web of Sience KeyUT A1991GN53800009
JaLCDOI 10.18926/AMO/30506
フルテキストURL fulltext.pdf
著者 Wato, Masaki| Shimomura, Hiroyuki| Fujio, Kozo| Tsuji, Hideyuki| Kondo, Junichi| Fujioka, Shin-ichi| Ishii, Yasushi| Hada, Hajime| Tsuji, Takao|
抄録 <p>We purified an apurinic/apyrimidinic (AP) endonuclease from mouse ascites sarcoma (SR-C3H/He) cells. The enzyme showed nicking activity on acid-depurinated DNA but not on untreated, intact DNA. It also showed priming activity for DNA polymerase on both acid-depurinated and bleomycin-damaged DNA. The priming activity on bleomycin-damaged DNA was two times higher than that on an acid-depurinated DNA. The enzymatic properties indicate that the enzyme is a class II AP endonuclease having DNA 3' repair diesterase activity. The purified enzyme has a molecular weight of 39,000 as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The optimal pH for AP endonuclease activity was 8.0 in 50 mM Tris-HCl buffer. The AP endonuclease activity depended on divalent cation such as Mg2+ and Co2+ ions, and was inhibited by 2 mM EDTA with no addition of the divalent cation. An appropriate concentration of sodium or potassium salt stimulated the activity. Partial digestion of the AP endonuclease with Staphylococcus aureus V8 protease produced 4 major peptide fragments which may be used for protein sequencing.</p>
キーワード hepatitis C ultracentrifugation immune complex interferon
Amo Type Article
発行日 1996-06
出版物タイトル Acta Medica Okayama
50巻
3号
出版者 Okayama University Medical School
開始ページ 139
終了ページ 144
ISSN 0386-300X
NCID AA00508441
資料タイプ 学術雑誌論文
言語 English
論文のバージョン publisher
査読 有り
PubMed ID 8805852
Web of Sience KeyUT A1996UU60400004
JaLCDOI 10.18926/AMO/30405
フルテキストURL fulltext.pdf
著者 Tamura, Tomoyuki| Koide, Norio| Hada, Hajime| Shiraha, Hidenori| Tsuji, Takao|
抄録 Adult rat hepatocytes assemble to form multicellular spheroids under non-adherent environments such as immobilized chondroitin sulfate-proteoglycan in primary culture. Previously, we demonstrated that hepatocyte spheroids exhibited various differentiated structures as observed in the liver tissue. It was also shown that hepatocyte growth was highly suppressed and several differentiated functions, including albumin production and gluconeogenesis, were well preserved in spheroids. To investigate the differentiated functions of cultured hepatocytes in relation to cell morphology, we compared the expression of the albumin and transferrin genes in spheroids with those in monolayers by Northern blot analysis. Production of these proteins in the culture medium was simultaneously examined by ELISA. Gene expression and protein production of both albumin and transferrin were better preserved in spheroids. We also examined changes in the expression of liver-specific genes in response to IL-6. Reduced mRNA levels of both albumin and transferrin was only found in spheroids and no change was observed in monolayers. These results suggest that the regulation of tissue-specific gene expression is better preserved in spheroids, in which hepatocytes are in close contact with each other.
キーワード hepatocyte spheroid primary culture gene expression IL-6
Amo Type Article
発行日 1995-06
出版物タイトル Acta Medica Okayama
49巻
3号
出版者 Okayama University Medical School
開始ページ 161
終了ページ 167
ISSN 0386-300X
NCID AA00508441
資料タイプ 学術雑誌論文
言語 English
論文のバージョン publisher
査読 有り
PubMed ID 7676847
Web of Sience KeyUT A1995RH05400007