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DEVELOPMENT OF RISK ASSESSMENT METHODS ON HAZARD IDENTIFICATION USING CATALASE MUTANT GENE AS A PREDICTIVE INDEX

Research Project

Project/Area Number 14570299
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Hygiene
Research InstitutionOKAYAMA UNIVERSITY

Principal Investigator

WANG Da-hong  Okayama University, Graduate School of Medicine and Dentistry, Assistant, 大学院・医歯学総合研究科, 助手 (90294404)

Co-Investigator(Kenkyū-buntansha) YAMAOKA Kiyonori  Okayama University, Medical School, Professor, 医学部, 教授 (00314683)
SANO Kuniaki  Okayama University, Graduate School of Medicine and Dentistry, Assistant, 大学院・医歯学総合研究科, 助手 (00294405)
TSUTSUI Ken  Okayama University, Advanced Science Research Center, Professor, 自然生命科学研究支援センター, 教授 (70108158)
KIRA Shohei  Okayama University, Graduate School of Medicine and Dentistry, Professor, 大学院・医歯学総合研究科, 教授 (50033212)
MASUOKA Noriyoshi  Okayama University, Graduate School of Medicine and Dentistry, Associate Professor, 大学院・医歯学総合研究科, 助教授 (20116502)
Project Period (FY) 2002 – 2003
Project Status Completed (Fiscal Year 2003)
Budget Amount *help
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2003: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 2002: ¥2,500,000 (Direct Cost: ¥2,500,000)
KeywordsCatalase mutant E coli strains / Acatalasemic mice / Hypocatalasemic mice / Catalase mutant genes / Hydrogen peroxide / Cytotoxicity / Antioxidants / Inorganic heavy metal compounds / アカタラセラミクスマウス / ヒポカタラセミクマウス / カタラーゼ阻害剤 / ニトロベンゼン / DNA酸化損傷
Research Abstract

This research was aimed at developing a risk assessment method for potential hazards by using catalase mutant strains. At first, we successfully constructed catalase mutant E coli strains by transforming plasmids encoding the catalase cDNA from catalase mutant mice (Cs^b, Cs^c) and its corresponding wild type (Cs^a) into catalase-deficient Escherichia coli UM255. There was a significant difference in catalase activities among each catalase-deficient E. coli mutant. The level of catalase in Csa appeared higher than that in the others, the catalase level in Cs^c was 65.1% of Cs^a, Cs^b was 61.7%, and UM 255 was 8.8% of Cs^a. Susceptibility to H_2O_2 showed a clear dose dependency with negative correlation to catalase activity levels. The susceptibility order was UM255>Cs^b>Cs^c>Cs^a. To assess the cytotoxicity of the chemicals, we employed colony-forming efficiency test and zones of inhibition test, the results demonstrated that survival of catalase-deficient E. coli mutants was signific … More antly reduced by treatment of hydroquinone, pyrogallol and L-Dopa in a dose-dependent manner and the cell viabilities underexposure to these chemicals were also different significantly among each catalase mutant strain, showing the following orders : Cs^a>Cs^c>Cs^b>UM255. The cytotoxicities induced by hydroquinone, pyrogallol and L-Dopa were also confirmed by zone of inhibition test, in which UM255 was the most susceptible, showing the largest zone of growth inhibition surrounding the paper disc, followed by Cs^b, Cs^c and Cs^a. The toxicities of these chemicals were completely blocked by catalase and ascorbic acid in a concentration-dependent relation. Treatment by some heavy metals (Hg, Cr, Cd, As, Ni, Fe, Cu) also significantly reduced the survival of these strains in a dose-dependent manner, suggesting H_2O_2 was increasingly produced in the process of hydroquinone cytotoxicity and catalase played a protective role in inhibiting the cell damage. The results support the usefulness of these newly established strains for hazard identification of oxidative chemicals in a risk assessment process. Less

Report

(3 results)
  • 2003 Annual Research Report   Final Research Report Summary
  • 2002 Annual Research Report
  • Research Products

    (14 results)

All Other

All Publications (14 results)

  • [Publications] Reiko Sunami: "Acatalasemia sensitizes renal tubular epithelial cells to apoptosis and exacerbates renal fibrosis following unilateral ureteral obstruction"Am J Physiol-Renal Physiol. (発表予定).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Noriyoshi Masuoka: "Characterization of hydrogen peroxide removal reaction by hemoglobin in the presence of reduced pyridine nucleotides."Biochim Biophys Acta. 1637. 46-54 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] 杉山 斉: "アカタラセミアマウスと酸化ストレス誘導腎障害"腎と透析. 54・6. 749-752 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Masako Horita, Da-hon Wang, Ken Tsutsui, Kuniaki Sano, Noriyoshi Masuoka, Shohei Kira: "Hydroquinone-induced cytotoxicity in catalase-deficient Escherichia coli mutants."Arch Toxicotol. (in submission).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Reiko Sunami, Hitoshi Sugiyama, Da-Hong Wang, Mizuho Kobayashi, Yohei Maeshima, Yasushi Yamasaki, Noriyoshi Masuoka, Norio Ogawa, Shohei Kira, Hirofumi Makino: "Acatalasemia sensitizes renal tubular epithelial cells to apoptosis and exacerbates renal fibrosis following unilateral ureteral obstruction."Am J Physiol-Renal Physiol. (in press).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Da-hong Wand, Noriyoshi Masuoka, Shohei Kira: "Animal model for oxidative stress research --Catalase mutant mice"Environ Health Prev Med. 8. 37-40 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Noriyoshi Masuoka, Hiroyuki Kodama, Tadashi Abe, Da-hon Wang, Taku Nakano: "Characterization of hydrogen peroxide removal reaction by hemoglobin in the presence of reduced pyridine nucleotides"Biochim Biophys Acta 1637. 1637. 46-54 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Hitoshi Sugiyama, Reiko Sunami, Da-Hong Wang, Shohei Kira, Hirofumi Makino: "Role of acatalasemia in the progression of oxidative stress-mediated renal injury."Kidney and Dialysis. 54(6). 749-752 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Reiko Sunami: "Acatalasemia sensitizes renal tubular epithelial cells to apoptosis and exacerbates renal fibrosis following unilateral ureteral obstruction"Am J Physiol-Renal Physiol. In press. (2004)

    • Related Report
      2003 Annual Research Report
  • [Publications] Noriyoshi Masuoka: "Characterization of hydrogen peroxide removal reaction by hemoglobin in the presence of reduced pyridine nucleotides"Biochim Biophys Acta. 1637. 46-54 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] 杉山 斉: "アカタラセミアマウスと酸化ストレス誘導腎障害"腎と透析. 54・6. 749-752 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Da-hong Wang: "Animal model for oxidative stress research-Catalase mutant mice"Environmental Health and Preventive Medicine. 8(2)(in press). (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] 堀田 昌子: "カタラーゼ遺伝子の変異を指標とした有害化学物質に対するリスク評価法の開発(第1報)"日本衛生学雑誌. 58(1). 197-197 (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] 益岡 典芳: "ジエチルニトロソアミン投与アカタラセミアマウスにおけるビタミンEの効果"第14回ビタミンE研究会プログラム・アブストラクト. 21-21 (2003)

    • Related Report
      2002 Annual Research Report

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Published: 2002-04-01   Modified: 2016-04-21  

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