• Journal of Microbiology and Biotechnology
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• v.20 no.11
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• pp.1563-1570
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• 2010

To construct a highly sensitive detection system for endocrine disruptors (EDs), we have compared the activity of promoters with the n-alkane-inducible cytochrome P450 gene (ALK1), isocitrate lyase gene (ICL1), ribosomal protein S7 gene (RPS7), and the translation elongation factor-1${\alpha}$ gene (TEF1) for the heterologous gene in Yarrowia lipolytica. The promoters were introduced into the upstream of the lacZ or hERa reporter genes, respectively, and the activity was evaluated by ${\beta}$-galactosidase assay for lacZ and Western blot analysis for hER${\alpha}$. The expression analysis revealed that the ALK1 and ICL1 promoters were induced by n-decane and by EtOH, respectively. The constitutive promoter of RPS7 and TEF1 showed mostly a high level of expression in the presence of glucose and glycerol, respectively. In particular, the TEF1 promoter showed the highest ${\beta}$-galactosidase activity and a significant signal by Western blotting with the anti-estrogen receptor, compared with the other promoters. Moreover, the detection system was constructed with promoters linked to the upstream of the expression vector for the hER${\alpha}$ gene transformed into the Y. lipolytica with a chromosome-integrated lacZ reporter gene under the control of estrogen response elements (EREs). It was indicated that a combination of pTEF1p-hER${\alpha}$ and CXAU1-2XERE was the most effective system for the $E_2$-dependent induction of the ${\beta}$-galactosidase activity. This system showed the highest ${\beta}$-galactosidase activity at $10^{-6}\;M\;E_2$, and the activity could be detected at even the concentration of $10^{-10}\;M\;E_2$. As a result, we have constructed a strongly sensitive detection system with Y. lipolitica to evaluate recognized/suspected ED chemicals, such as natural/synthetic hormones, pesticides, and commercial chemicals. The results demonstrate the utility, sensitivity, and reproducibility of the system for identifying and characterizing environmental estrogens.

• Development and Reproduction
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• v.13 no.3
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• pp.163-172
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• 2009

Some organotin compounds such as butyltins and phenyltins are known to induce impo-sex in various marine animals and are considered to be endocrine disruptors. In this study, the effect of organotins on follicular steroidogenesis in amphibians was examined using ovarian follicles of Rana dybowskii and Rana catesbeiana. Isolated follicles were cultured for 6 or 18 h in the presence and absence of frog pituitary homogenate (FPH) or various steroid precursors, and the levels of product steroids in the culture media oassay. Among the butyltin compounds, tributyltin (TBT) strongly and dose-dependently inhibited the FPH-induced synthesis of pregnenolone ($P_5$) and progesterone ($P_4$) by the follicles. TBT also strongly suppressed the conversion of cholesterol to $P_5$ and partially suppressed the conversion of $P_5$ to $P_4$. A high concentration of dibutyltin (DBT) also inhibited steroidogenesis by the follicles while monobutyltin and tetrabutyltin had negligible effects. The toxic effect of TBT or DBT was irreversible and a short time of exposure (30 min) was enough to suppress steroidogenesis. All the phenyltin compounds significantly inhibited FPH-induced $P_5$ synthesis by the follicles. The effective dose of 50% inhibition by diphenyltin was $0.04\;{\mu}M$ and those of monophenyltin and triphenyltin were $0.24\;{\mu}M$ and $0.3\;{\mu}M$, respectively. However, none of the phenyltin compounds significantly suppressed the conversion of $P_4$ to $17{\alpha}$-hydroxyprogesterone ($17{\alpha}$-OHP) (by $17{\alpha}$-hydroxylase), $17{\alpha}$-OHP to androstenedione (AD) (by $C_{17-20}$ lyase), or AD to testosterone by the follicles. Taken together, the data show that among the steroidogenic enzymes, P450scc in the follicles is the most sensitive to organotin compounds and that an amphibian follicle culture system can be a useful screening model for endocrine disruptors.

• Journal of Food Hygiene and Safety
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• v.14 no.4
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• pp.408-414
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• 1999

A number of chemical released into the environment eliciting their effects by disrupting normal hormonal pathways. Endocrine disrupting compounds are present in the aquatic environment and pose potential health consequences to wildlife and humans. This review are designing fur xenobiotic estrogens based on induction of the egg-yolk precursor protein vitelloge-nin. In fish of aquatic environment, it may result in decrease fertility and egg production in females or lead to reduced gonad size or feminization of genetic male fish. It has been known that male fish exposed to estrogenic compounds show induced production of vitellogenin. Vitello-genin production is normally restricted to adult females, which have elevated estrogen levels during egg production. However, vitellogenin can be induced in males by Pollution of environmental endocrine disruptors. Consequently, the presence of vitellogenin in male fish can serve as an indicator of exposure of environmental endocrine disrupting compounds. In immature fish pol-luted at low levels of environmental endocrine disrupter, vitellogenin can serve as a reliable biomarker for exposure to endocrine disrupter. This review demonstrates the utility of vitellogenin as a biomarker fur exposure to estrogenin agents in auqatic environment.

• Proceedings of the Korea Society of Environmental Toocicology Conference
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• 2000.12a
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• pp.56-56
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• 2000

• Proceedings of the Korea Environmental Mutagen Society Conference
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• 2002.11a
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• pp.169-169
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• 2002

• Proceedings of the Korea Environmental Mutagen Society Conference
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• 2003.05a
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• pp.62-63
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• 2003

• Proceedings of the Korea Environmental Mutagen Society Conference
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• 2003.05a
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• pp.78-79
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• 2003

• Proceedings of the Korean Society of Food Hygiene and Safety Conference
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• 2001.10a
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• pp.264-266
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• 2001

• Proceedings of the Korean Society of Developmental Biology Conference
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• 2004.07a
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• pp.33-33
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• 2004

• Proceedings of the Korean Society of Toxicology Conference
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• 2003.05a
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• pp.62-63
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• 2003

A case-control study was designed to determine the possible association between dioxin like compounds (such as TCDD, PCDDs, PCDFs, and PCBs) and the occurrence and severity of endometriosis using CALUX bioassay method.(omitted)