DEVELOPMENT AND ITS PRACTICAL USE OF SCREENING METHODS ON HAZARD IDENTIFICATION USING CATALASE MUTANT GENE AS A PREDICTIVE INDEX

• Project/Area Number: 17390173
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Hygiene
• Research Institution: Okayama University
• Principal Investigator: WANG Da-hong Okayama University, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Dentistry Sciences Assistant Professor (90294404)
• Co-Investigator(Kenkyū-buntansha): TSUTSUI Ken Okayama University, Advanced Science Research Center, Professor (70108158) ; SANO Kuniaki Okayama University, Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, Assistant Professor (00294405) ; MASUOKA Noriyoshi Okayama University of Science, Department of Life Science, Professor (20116502) ; ITO Takehiko Okayama University, Faculty of Education, Professor (10291973) ; OGINO Keiki Okayama University, Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, Dentistry Sciences, Professor (70204104)
• Project Period (FY): 2005 – 2007
• Project Status: Completed (Fiscal Year 2007)
• Budget Amount: ¥7,520,000 (Direct Cost: ¥7,100,000、Indirect Cost: ¥420,000); Fiscal Year 2007: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2006: ¥2,700,000 (Direct Cost: ¥2,700,000); Fiscal Year 2005: ¥3,000,000 (Direct Cost: ¥3,000,000)
• Keywords: Catalase mutant gene / Transformants / CAT assay / Chemical compounds / Risk assessment / Oxidative stress / Antioxidants / Screening / ジチオトレイトール / 過酸化水素 / ローソン / ヘンナ製品 / ヒドロキノン / クメンハイドロペルオキサイド / 浸出水と放流水

Research Abstract

This research aimed at developing a screening method for hazard identification using catalase mutant E coli strains constructed by encoding the catalase cDNA from catalase mutant mice (Cs^b) and its corresponding wild type (Cs^a) into catalase-deficient K coli UM255. To assess the cytotoxicity of the chemicals, we employed a CAT assay (a zone of inhibition test of bacterial growth and colony-forming efficiency test using catalase mutant E coli strains). Lawsone exposure inhibited the growth of both Cs^a and Cs^b strains in a dose-dependent manner, in which Cs^b was more susceptible than Cs^a, showing the larger zone of growth inhibition surrounding the paper disc. Natural henna leaves did not show toxic effects, whereas two out of four samples of market henna products were shown toxicity effects. Catalase abolished zone of inhibition of henna products, and low concentration of exogenous MnSOD and Cu/ZnSOD eliminated the toxicity. We also exposed dithiothreitol (DTT) to catalase mutant E coli strains. The results show that DTT was also toxic to the catalase mutant K colis in dose dependent manner and significantly different between Cs^a and Cs^b. Catalase, CuZnSOD, and MnSOD also showed protective ability on tester strains. Treatment with waste disposal extracted by methanol and sterilized water showed a negative relation between the exposure dose and survival of these strains. The results support the usefulness of these newly established strains for hazard identification of oxidative chemicals in a risk assessment process.

Research Products

• [Journal Article] Development of a new assay for evaluation of L-3, 4-dihydroxyphenylalanine cytotoxicity in catalase mutant Escherichia coli. (2008)
  • Author(s): Wang DH, Horita M, Tsutsui K, Sano K, Sauriasari R, Masuoka N, Takemura Y, Takigawa T, Takaki J, Ogino K.
  • Journal Title: Environmental Toxicology and Chemistry (In press)
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Development of a new assay for evaluation of L-3,4-dihydroxy- phenylalanine cytotoxicity in catalase mutant Escherichia coli (2008)
  • Author(s): Wane, DH, Horita, M., Tsutsui, K., Sano, K., Sauriasari, R., Masuoka, N., Takemura, Y., Takigawa, T., Takaki, J., Ogino, K
  • Journal Title: Environmental Toxicology and Chemistry (in press)
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Development of a new assay for evaluation of L-3, 4-dihydroxyphenylalanine cytotoxicity in catalase mutant Escherichia coli. (2008)
  • Author(s): Wang DH, Horita M, Tsutsui K, Sano K. Sauriasari R, Masuoka N, Takemura Y, Takigawa T. Takaki J. Ogino K.
  • Journal Title: Environmental Toxicology and Chemistry (in press)
  • Peer Reviewed
• [Journal Article] Cytotoxicity of lawsone and cytoprotective activity of antioxidants in catalase mutant Escherichia coli. (2007)
  • Author(s): Sauriasari R, Wang DH, Takemura Y, Tsutsui K, Masuoka N, Sano K, Horita M, Wang BL, Ogino K.
  • Journal Title: Toxicology 235 Pages: 103-111
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Cytotoxicity of lawsone and cytoprotective activity of antioxidants in catalase mutant Escherichia coli (2007)
  • Author(s): Sauriasari, R., Wang, DH, Takemura, Y., Tsutsui, K., Masuoka, N., Sano, K., Horita, M., Wang, BL, Ogino, K
  • Journal Title: Toxicology 235 Pages: 103-111
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Cytotoxicity of lawsone and cytoprotective activity of antioxidants in catalase mutant Escherichia coli (2007)
  • Author(s): Sauriasari R, Wang DH, Takemura Y, Tsutsui K, Masuoka N, Sano K, Horita M. Wang BL, Ogino K
  • Journal Title: Toxicology 235 Pages: 103-111
  • Peer Reviewed
• [Journal Article] Protective ability of enzymatic and non enzymatic antioxidants against dithiothreitol-induced cytotoxicity on catalase mutant Escherichia coli (2007)
  • Author(s): Sauriasari R, Wang DH, Takemura Y, Tsutsui K, Masuoka N, Sano K, Wang BL, Horita M, Ogino K
  • Journal Title: Journal of Preventive Medicine 2 Pages: 36-41
  • NAID: 40016137834
  • Peer Reviewed
• [Journal Article] Cytotoxicity of lawsone and cytoprotective activity of antioxidants in catalase mutant Escherichia coli. (2007)
  • Author(s): Sauriasari R, Wang DH, Takemura Y, Tsutsui K, Masuoka N, Sano K, Horita M, Wan BL, 0gino K.
  • Journal Title: Toxicology (In press)
• [Journal Article] Characterization of acatalasemic erythrocytes treated with low and high dose hydrogen peroxide: Hemolysis and aggregation. (2006)
  • Author(s): Masuoka N, Sugiyama H, Ishibashi N, Wang DH, Masuoka T, Kodama H, Nakano T.
  • Journal Title: The Journal of Biological Chemistry 281 Pages: 21728-21734
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Characterization of acatalasemic erythrocytes treated with low and high dose hydrogen peroxide : Hemolysis and aggregation (2006)
  • Author(s): Masuoka, N., Sugiyama, H., Ishibashi, N., Wang, DH, Masuoka, T., Kodama, H., Nakano, T
  • Journal Title: The Journal of Biological Chemistry 281 Pages: 21728-21734
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Characterization of acatalasemic erythrocytes treated with low and high dose hydrogen peroxide. Hemolysis and aggregation. (2006)
  • Author(s): Masuoka N, Sugiyama H, Ishibashi N, Wang DH, Masuoka T, Kodam H, Nakano T
  • Journal Title: The Journal of Biological Chemistry 281
• [Journal Article] Involvement of oxidative stress in hydroquinone-induced cytotoxicity in catalase-deficient Escherichia coli mutants. (2005)
  • Author(s): Horita M, Wang_DH, Tsutsui K, Sano K, Masuoka N, Kira S.
  • Journal Title: Free Radical Research 39・10 Pages: 1035-1041
• [Journal Article] Catalase deficiency renders remnant kidneys more susceptible to oxidant tissue injury and renal fibrosis in mice. (2005)
  • Author(s): Kobayashi M, Sugiyama H, Wang DH, Toda N, Maeshima Y, Yamasaki Y, Masuoka N, Yamada M, Kira S, Makino H.
  • Journal Title: Kidney International 68・3 Pages: 1018-1031
• [Presentation] カタラーゼ変異遺伝子を導入した大腸菌におけるローソンによる細胞毒性と抗酸化物質の抑制作用。 (2007)
  • Author(s): ラニ サウリアサリ、汪達紘、竹村洋子、筒井研、益岡典芳、佐野訓明、筒井公子、堀田昌子、王炳玲、荻野景規
  • Organizer: 第18回中国四国生体ラジカル研究会
  • Place of Presentation: 岡山理科大学
  • Year and Date: 2007-07-20
• [Presentation] 染色剤に含まれるピロガロールの細胞毒性と活性酸素の関与 (2007)
  • Author(s): 竹村洋子、汪達紘、ラニ サウリアサリ、堀田昌子、益岡典芳、伊藤武彦、荻野景規
  • Organizer: 第77回日本衛生学会総会
  • Place of Presentation: 大阪国際交流センター
  • Year and Date: 2007-03-28
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] Involvement of oxidative stress in pyrogallol-induced mutagenicity and cytotoxicity (2007)
  • Author(s): Takemura, Y., Wang, DH, Sauriasari, R., Horita, M., Masuoka, N., Ito, T., Ogino, K
  • Organizer: The 77th annual meeting of the Japanese Society for Hygiene
  • Place of Presentation: Osaka
  • Year and Date: 2007-03-28
  • Description: 「研究成果報告書概要(欧文)」より