• Title/Summary/Keyword: Ethoxyresorufin O-deethylase

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Comparative Study of TCDD Effect on CYP1A1 Gene Expression in Trout and Mouse Liver Cells (송어 간세포와 생쥐 간세포에서 TCDD가 CYP1A1 유전자 발현에 미치는 영향 비교 연구)

  • Kim, Ji-Sun;Min, Kyung-Nan;Sheen, Yhun-Yhong
    • Environmental Mutagens and Carcinogens
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    • v.24 no.3
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    • pp.143-150
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    • 2004
  • In mammalian, cytochrome P4501A1 (CYP1A1) is very important for metabolism of xenobiotics such as PAHs(Polycyclic aromatic hydrocarbon) and heterocyclic amine, and it is induced by environmental contaminants such as PAHs, TCDD(2,3,7,8-tetrchlorodibenzo-p-dioxin) and 3-MC (3-methylcholanthrene). In fish, like mammalian, when it is exposed to environmental contaminants, they cause specific and sensitive induction of CYP1A. Therefore, induction of CYP1A in fish and mammalian is widely used as a biomarker for exposure of environmental contaminants. In this study, to compare the function of Cyp1a1 in fish with it in mammalian, we have used rainbow trout(Oncorhynchys mykiss) hepatoma cells (RTH-149) and mouse hepatocyte (Hepa-I). in order to examine induction of Cyp1a1 by TCDD, we have used the bioassay system. We examined effects of TCDD on the Cyp1a1-luciferase reporter gene activity, 7-ethoxyresorufin O-deethylase(EROD) activity and Cypa mRNA level.

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Determination of Dioxin-like Components in the School Waste Incinerator Residues by EROD-microbioassay (EROD-microbioassay에 의한 학교 소각로 잔재 중 다이옥신 유사물질의 측정)

  • 정규혁;오승민;윤완진
    • Journal of Environmental Health Sciences
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    • v.26 no.3
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    • pp.11-17
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    • 2000
  • There are among the most relevant toxic emissions from incinerators such as polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs), dioxin-like polychlorinated biphenyls (PCBs). Induction of cytochrome P4501A1 catalyzed 7-ethoxyresorufin O-deethylase(EROD) activity in mammalian cell culture(EROD bioassay) is thought to be a selective and sensitive parameter used for the quantification of dioxin-like components. In this study, the toxic emissions from several school waste incinerators were evaluated by determination of CYPIA catalytic activity and cytotoxicity using cell culture microbioassay. The incinerator residue and soil samples were collected from the schools located in Kyunggi province from April to June 1999. The samples were extracted in a Soxhlet apparatus using toluene for 20 hours. In order to clean-up, concentrated crude extracts were applied to basic alumina column. The EROD activities of extracts in the H4IIE cells were from 1.91$\pm$0.32 ng-TEQ/g to 24.54$\pm$3.48 ng-TEQ/g of biochemical-TEQ value. In soil samples, CYP1A catalytic activity was 0.09~0.64 ng-TEQ/g. EROD bioassay, seems to be a useful short-term bioassay when information about the biological response of complex environmental samples is needed. Although further study is needed, these results indicate that the potent toxic emissions are produced from school waste semi-incinerators.

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Cell Culture Microbioassay for the Water Pollution Monitoring (세포배양 생화학적 기법에 의한 수환경오염 평가)

  • 오승민;정규혁
    • Toxicological Research
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    • v.16 no.4
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    • pp.285-291
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    • 2000
  • So far, investigation of environmental pollution has been achieved in field study. This remains the most exhaustive approach, current dimensions of environmental researches and their inherent complexity require that relatively inexpensive and simple laboratory procedures are developed to make possible the screening of large numbers of sites and samples. At this point. microbioassay has been high-lighted. The purpose of this study is to evaluate the water pollution using microbioassay. Two microbioassay methods were optimized and validated for the sensitive and quantitative determination of total toxic effects in the water. EROD(Ethoxyresorufin-O-deethylase) microbioassay was focused to detect PARs, PCBs and dioxinlike components in the water and E-screen assay to xenoestrogens. The EROD microbioassay was executed in rat hepatoma cell line, H4IIE and E-screen assay in MCF7-BUS cell line. Kumho river was selected for this study. 5ι of river water was extracted using combined solid-phase extraction in static adsorption mode with soxhlet extraction. Pollutants adsorbed to the XAD-4 resin were recovered by elution with ethyl acetate and methylene chloride (1 : 9). Toxic effects of extracts were determined by EROD-microbioassay and E-screen assay. EROD activities of water samples were 7.24-72.24 ng/ι MEQ. The estrogenic effect of various water samples is quantitatively evaluated by EEQ. The EEQ of samples range from 0.05 to 6.07 ng-EEQ/ι. These results suggested that Kumho river was highly polluted with organic toxic chemicals.

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Comparison of Xenobiotic Metabolism and Antioxidative Status in Cultivated and Stocked Rockfish Sebastes schlegeli (양식산과 방류산 조피볼락(Sebastes schlegeli)의 간장 중 해독 및 항산화 효소계의 비교)

  • Lee, Ji-Seon;Byun, Hee-Guk;Jeon, Joong-Kyun
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.45 no.2
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    • pp.97-103
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    • 2012
  • This study compared the composition and texture of muscle and levels of detoxification and antioxidant enzymes in the livers of stocked and cultivated rockfish Sebastes schlegeli released after the primary culture stage in Tongyoung, South Korea. The crude lipid content of muscle was significantly higher ($P$<0.05) in cultivated rockfish than stocked rockfish, while the texture did not differ significantly ($P$>0.05). The condition factor (CF) and hepatosomatic index (HSI) did not differ significantly and the growth of stocked and cultivated rockfish was similar. The levels of the detoxification enzymes cytochrome P450 (CYP) and Ethoxyresorufin-O-deethylase (EROD) were significantly lower in the livers of stocked rockfish, perhaps because of their reduced exposure to xenobiotic compounds. In addition, stocked rockfish had a significantly ($P$<0.05) lower CAT and higher GR than cultivated rockfish, but similar levels of tGPx, SOD, GSH, and GSSG. The present study shows that the growth rates of stocked and cultivated rockfish are similar and that stocked rockfish are exposed to fewer xenobiotic compounds and less oxidative stress.

Effect Evaluation of Benzo[a]pyrene on Multiple Biomarkers in Common Carp (Cyprinus carpio) (잉어 (Cyprinus carpio)의 다중바이오마커를 이용한 Benzo[a]pyrene의 영향평가)

  • Kim, Woo-Keun;Kim, Ja-Hyun;Yeom, Dong-Hyuk;Lee, Sung-Kyu
    • Environmental Analysis Health and Toxicology
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    • v.23 no.3
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    • pp.171-178
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    • 2008
  • 수백여 종의 개별물질이 불완전 연소 혹은 유기물의 열분해로 인해 발생되는 다환방향족 탄화수소(PAHs)는 환경에서 중요한 오염원이 되고 있다. 본 연구는 다양한 바이오마커를 이용하여 수서생태계에 벤조피렌(benzo[a]pyrene)과 같은 다환방향족 탄화수소의 영향을 분석하였고, 이에 대한 통합적 결과 모델을 도출하였다. 즉, 잉어(Cyprinus carpio)를 이용하여 여러 농도의 벤조피렌(3, 12, $34{\mu}g/L$, 측정농도 기준)에 10일간 노출시킨 다음, DNA single-strand break, ethoxyresorufin-O-deethylase (EROD), acetylcholine esterase (AChE)와 vitellogenin (VTG)의 농도를 측정하였다. 벤조피렌은 잉어의 DNA 손상을 유도하였고, 낮은 농도에서 EROD와 VTC의 유의적인 활성을 보였으나, 신경전달물질과 관련이 깊은 AChE 효소활성에는 영향을 미치지 않았다. 이 결과를 star plot를 이용하여 통합 및 분석하였으며, 노출농도에 따른 통합 반응지수(integrated biomarker response value: IBR)로 나타내었다. 이런 다양한 바이오마커의 결과들은 벤조피렌에 대한 어류의 영향과 수생태 모니터링 자료로 이용 가능할 것으로 여겨지며, 통합반응지수는 생태위해성평가에서 유용한 도구로 쓰일 가치가 있는 것으로 평가된다.

Studies on Toxicological Evaluation of Freshwater Sediment using a PLHC-1 Cell Comet Assay (PLHC-1세포주의 Comet assay를 이용한 하천 퇴적토의 생태독성평가)

  • Bak, Jeong-Ah;Hwang, In-Young;Baek, Seung-Hong;Kim, Young-Sug
    • Korean Journal of Environmental Biology
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    • v.29 no.1
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    • pp.23-30
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    • 2011
  • In this study, the Comet assay (evaluation of DNA damage) used the fish hepatocellular carinoma cell, PLHC-1, was tried to the sediment extract obtained from freshwater to understand its applicability as a tool for monitoring sediment toxicity. In parallel, induced EROD (7-ethoxyresorufin- O-deethylase) activity and DNA damage (TEM values) in PLHC-1 cells were measured for establishing the tandem endpoints of the PLHC-1cell test to test the ecotoxicity of sediment. Among several study sites in a small river passed through downtown and industrial park area, one of them, site B, showed a higher level of EROD activity and DNA damage than other sites. It indicates that a tandem endpoints of PLHC-1 cells could be useful tools for assessing the toxicity of sediment. The sensitivity of Comet assay with PLHC-1 cells was a little higher than that with a blood cell of frog tadpoles to the solvent extract of sediment. According to the results, a PLHC-1 cell-Comet assay could be used as a useful tool for evaluating ecotoxicity of the freshwater sediment. In addition, more detailed studies are needed to the contaminated site.

Effect of PAH on CYP1A1 Gene Expression in Trout RTH-149 Cells (PAH가 송어 RTH-149세포에서 CYP1A1 유전자 발현에 미치는 영향)

  • Kim, Ji-Sun;Sheen, Yhun-Yhong
    • Environmental Mutagens and Carcinogens
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    • v.24 no.4
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    • pp.155-162
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    • 2004
  • In mammalian, cytochrome P4501A1 (CYP1A1) is very important for metabolism of xenobiotics such as PAHs(Polycyclic aromatic hydrocarbon) and heterocyclic amine, and it is induced by environmental contaminants such as PAHs, TCDD(2,3,7,8-tetrchlorodibenzo-p-dioxin) and 3-MC (3-methylcholanthrene). In fish, like mammalian, when it is exposed to environmental contaminants, they cause specific and sensitive induction of CYP1A. Therefore, induction of CYP1A in fish and mammalian is widely used as a biomarker for exposure of environmental contaminants. In this study, to compare the function of Cyp1a1 in fish with it in mammalian, we have used rainbow trout(Oncorhynchys mykiss) hepatoma cells (RTH-149) and mouse hepatocyte (Hepa-I). in order to examine induction of Cyp1a1 by TCDD, we have used the bioassay system. We examined effects of TCDD on the Cyp1a1-luciferase reporter gene activity, 7-ethoxyresorufin O-deethylase(EROD) activity and Cypa mRNA level.

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Induction of Imposex in Rock Shell, Thais clavigera, Exposed to Organotins and Other Endocrine Disrupting Chemicals

  • Shim, Won Joon
    • Proceedings of the Korea Society of Environmental Biology Conference
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    • 2003.11a
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    • pp.7-11
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    • 2003
  • Interactions of both feminizing and masculinizing chemicals inducing imposex in gastropod were investigated with a long-term exposure experiment. Imposex-free rock shell(Thais-clavigera)was exposed to TBT and other antagonistic chemicals (nonylphenol and DDT), P450 inducer (phenobarbital) and natural sex hormone (estrogen) mixtures for 60 days. None of imposexed-females were found in control, sham (ethanol) and testosterone exposure group, on the other hand imposex was induced in the TBT and all the TBT+antagonist mixture groups. After 60-day exposure, frequency of imposexed-female ranged from 31-68%. Female mean penis length (FMPL) of imposexed-female ranged from 0.87-2.58 mm, and relative penis length indices were from 7.2 to 21.2%. However, the degrees of imposex were different among the exposure groups. The FMPLS of three exposure groups (TBT+estrogen, + DDT and +nonylphe- not) were less than that of the TBT exposure group. Ethoxyresorufin-O-deethylase (EROD) activities in gastropod decreased in the TBT and all the TBT+antagonist ekposure groups compared to the control group. A negative correlation was abtained between TBT body residue and EROD activity, while a positive relationship was obtained between 737 body residue and the degree of imposex except for the TBT +DDT exposure group. Although 737 concentration was relatively high in the TBT+DDT exposure group, the TBT +DDT exposure group demonstrated low EROD activity and low degree of imposex. These results indicate that a certain antagonistic effect of DDT occurred in induction of imposex by TBT.

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Antiestrogenic Effects of Marijuana Smoke Condensate and Cannabinoid Compounds

  • Lee Soo Yeun;Oh Seung Min;Lee Sang Ki;Chung Kyu Hyuck
    • Archives of Pharmacal Research
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    • v.28 no.12
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    • pp.1365-1375
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    • 2005
  • The antiestrogenic effects of marijuana smoke condensate (MSC) and three major cannabinoids, i.e., $\bigtriangleup^{9}$-tetrahydrocannabinol (THC), cannabidiol (CBD), and cannabinol (CBN), were evaluated using in vitro bioassays, viz., the human breast cancer cell proliferation assay, the recombinant human estrogen receptor (ER) competitive binding assay, and the reporter gene assay. The inhibitory effects on estrogen were also examined using the ethoxyresorufin-O­deethylase (EROD) assay, the aromatase assay, and the 17$\beta$-estradiol ($E_{2}$) metabolism assay. The results showed that MSC induced the antiestrogenic effect via the ER-mediated pathway, while THC, CBD, and CBN did not have any antiestrogenic activity. This suggests that the combined effects of the marijuana smoke components are responsible for the antiestrogenicity of marijuana use. In addition, MSC induced the CYP1A activity and the $E_{2}$ metabolism, but inhibited the aromatase activity, suggesting that the antiestrogenic activity of MSC is also related to the indirect ER-dependent pathway, as a result of the depletion of the in situ $E_{2}$ level available to bind to the ER. In conclusion, pyrogenic products including polycyclic aromatic hydrocarbons (PAHs) in the non-polar fraction, which is the most biologically active fraction among the seven fractions of MSC, might be responsible for the antiestrogenic effect.

Effects of Trialkyltin in vitro on the Microsomal Monooxygenase System of Digestive Gland in the Clam, Coelomactra antiquata (유기주석화합물이 명주조개 (Coelomactra antiquata)의 약물대사효소계에 미치는 영향)

  • Jeon Joong Kyun;Lee Mee Hee;Kim Do Jin;Shim Won Joon;Oh Jae Ryong;Lee Soo Hyung
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.35 no.2
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    • pp.185-190
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    • 2002
  • This study was carried out to measure the in vitro interaction of trialkyltin with the microsomal monooxygenase (MFO) system of the clam, Coelomactra antiquata. Cytochrome P450 (CYP) level and 7-ethonvesorufin-O-deethylase (EROD) activity were invetigated in the microsome isolated from digestive gland of the clam (C. antiquata) exposed to tributyltin chloride (TBTC), bis-tributyltinoxide (TBTO) and triphenyltin chloride (TPTC). The specific contents of CYP in clam microsome exposedto 0.4 mM TBTC, TBTO and TPTC for 20 minutes were decreased 52, 72 and $40\%$, respectively, compared to control group. The EROD activities also were inhibited by exposure to TBTO ($92\%$) and TPTC ($85\%$) except for TBTC, The level of CYP and the EROD actintles were decreased according to the OTC exposure concentrations. The toxic effects on the level of CYP and the EROD activities were in order of TPTC>TBTC>TBTO in this study. The measurement of CYP level and EROD activity could be applied as a biomarker for environmental study.