• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2005년도 생물공학의 동향(XVI)
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• pp.88-92
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• 2005

Aptamer is the single-stranded oligonucleotide which binds to various target molecules such as proteins, peptides, lipids and small organic molecules with high affinity and specificity. DNA aptamers specific for the $17{\beta}-estradiol$ were selected by SELEX (Systematic Evolution of Ligands by EXponential enrichment) process from a random DNA library. These DNA aptamers have a high affinity to $17{\beta}-estradiol$ as an endocrine disrupting chemical. Nanowell and $200{\mu}m$ gold electrode were used as substrate for DNA aptamer immobilization and electrochemical analysis. Especially, nanowell gold electrode was fabricated by e-beam lithography. The size of single nanowell is 130nm and 40,000 nanowells were deposited on one gold electrode. The immobilization method was based on the interaction between the biotinylated aptamer and streptavidin deposited on gold electrode previously. Immobilization procedure was optimized by surface plasma resonance (SPR) and electrochemical analysis. After the immobilization of DNA aptamer on streptavidin modified gold electrode, $17{\beta}-estradiol$ solution was treated on aptamer immobilized gold electrode. The current of gold electrode was decreased by the binding of $17{\beta}-estradiol$ to DNA aptamer immobilized on gold electrode. However, in negative control experiments of 1-aminoanthraquinone and 2-methoxynaphthalene, the current was rarely decreased. And more sensitive data was obtained from nanowell gold electrode comparing with $200{\mu}m$ gold electrode.

• Clinical and Experimental Reproductive Medicine
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• 제39권1호
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• pp.15-21
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• 2012

Objective: Tributyltin (TBT), an endocrine disrupting chemical, has been reported to decrease ovarian function by causing apoptosis in the ovary, but the mechanism is not fully understood. Therefore, we examined whether TBT increases the expression of adipogenesis-related genes in the ovary and the increased expression of these genes is associated with apoptosis induction. Methods: Three-week-old Sprague-Dawley rats were orally administered TBT (1 or 10 mg/kg body weight) or sesame oil as a control for 7 days. The ovaries were obtained and weighed on day 8, and then they were fixed for terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) or frozen for RNA extraction. Using the total RNA of the ovaries, adipogenesis- and apoptosis-related genes were analyzed by real-time polymerase chain reaction (PCR). Results: The ovarian weight was significantly decreased in rats administered 10 mg/kg TBT compared to that in control rats. As determined by the TUNEL assay, the number of apoptotic follicles in ovary was significantly increased in rats administered 10 mg/kg TBT. The real-time PCR results showed that the expression of adipogenesis-related genes such as $PPAR{\gamma}$, ${\alpha}P2$, CD36, and PEPCK was increased after TBT administration. In addition, apoptosis-related genes such as $TNF{\alpha}$ and TNFR1 were expressed more in the TBT-administered rats compared with the control rats. Conclusion: The present study demonstrates that TBT induces the expression of adipogenesis- and apoptosis-related genes in the ovary leading to apoptosis in the ovarian follicles. These results suggest that the increased expression of adipogenesis-related genes in the ovary by TBT exposure might induce apoptosis resulting in a loss of ovarian function.

• 한국환경보건학회지
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• 제29권2호
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• pp.16-22
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• 2003

BBP (Butyl benzyl phthalate), a widely used plasticizer. can enter the food and environment as consequence of its manufacture, use, and disposal. BBP was found to be developmental and teratogenic or endocrine disrupting chemical in rats. The effects of BBP were investigated in female rats (P) and second generation (F1) via lactations. Sprague-Dawley were given BBP by oral administration at 0, 5, 10, 100, 1000 mg/kg on day 0 to 21 of lactation period. The results were as follows : At maternal findings, there were some significant changes (p<0.05) in relative organ weight, especially liver and uterus weight by BBP administration. In estrous cycle, high treated group was inclined to be proestrus or estrus compared to control group. BBP indues estrous cycle earlier than the control group. At fetal findings, there were some significant changes in relative liver and spleen weight, especially 100, 1000 mg/kg administered groups. The relative weight of ventral prostate was decreased, so it was represent to dose-response tendency. Parent rats (P) were detected monobenzyl phthalate (MBeP) 3.21~5.81 $\mu\textrm{g}$/ml in 100, 1000 mg/kg dose groups. MBeP of male and female fetuses (F1) were detected at the level of 1.21~2.63 $\mu\textrm{g}$/ml of serum. Male serum concentration oi MBeP was higher than the females'. Estrogen receptor $\alpha$ expression by BBP and bisphenol A in uterus and testis of F1 were studied. The ER$\alpha$ expression were increased in F1 male testis and female uterus. F1 male showed distint ER$\alpha$ expression, especially in the combined exposrue. Synergistic ER$\alpha$ expression was found by combined treatment group of BBP and bisphenol A. From the above results, it could be concluded that the effects of dams and F1 by BBP administration during lactation period were estrogenic, and BBP can transfer to F1 via lactation, and make estrogenic at F1 reproductive organs.

• Biomolecules & Therapeutics
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• 제8권1호
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• pp.78-83
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• 2000

The objective of this study was to prevalidate the Organization for Economic Cooperation and Development's (OECD) rodent uterotrophic assay as a test method for screening of potential endocrine disrupting chemicals (EDCs). This study was conducted exactly as described in the OECD protocol documents. A positive control substance, 17$\alpha$-ethinyl estradiol (EE), was administered daily for three days to ovariectomized (OVX) Sprague-Dawley rats at various doses for determine the dose-response curve. Additionally, a pure antiestrogenic chemical, ZM189, 154 was administered to OVX rats at the same time EE to determine the effectiveness of the material against blocking the estrogenic effects of EE. At higher concentration of EE (10 $\mu\textrm{g}$/kg), a statistically significant difference in body weight gain and food consumption was observed compared to vehicle controls. In uterine responses, EE produced a dose-related increase in uterus weights compared to vehicle control. These increases were statistically significant at the >1.0 $\mu\textrm{g}$/kg doses. However, a similar dose-response relationship was not observed in vagina weight. A comparison of the two groups receiving ZM189,154 (0.1 and 1.0 mg/kg) with 0.3 $\mu\textrm{g}$/kg of EE and the group receiving only 0.3 $\mu\textrm{g}$/kg of EE showed dose-related decreases in uterus weights. However, statistical significance was shown in 1.0 mg/kg of ZM189,154. In conclusion, administration of EE produced a dose-related increase in uterine (wet and blotted) weights. Additionally, the 1.0mg/kg dose of ZM189,154 was effective in blocking the estrogenic activity of EE. These data suggest 3-day uterotrophic assay using OVX rats may serve as a good tool for EDCs screening.

• 한국환경독성학회:학술대회논문집
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• 한국환경독성학회 2003년도 추계국제학술대회
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• pp.173-173
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• 2003

With the introduction of DNA microarrays, a high throughput analysis of gene expression is now possible as a replacement to the traditional time-consuming Southern-blot analysis. This cDNA microarray should be ahighly favored technology in the area of molecular toxicology or analysis of environmental stresses.In this study, therefore, we developed a novel cDNA microarray for analyzing stress-specific responses in japanese Medaka fish. In the design and fabrication of this stress specific functional cDNA microarray, 123 different genes in Medaka fish were selected from eighteen different stress responsive groups and spotted on a 25${\times}$75 mm glass surface. After exposure of the fish to bisphenol A which is the one of the well-known endocrine disrupting chemicals (EDCs), over 1 or 10 days, the responses of the DNA chip were found to show distinct expression patterns according to the mode of toxic actions from environmental toxicants. As a results, they showed specific gene expression pattern to bisphenol A, additionally, the chemical spesific biomarkers could be suggested based on the chip analysis data. Therefore, this chip can be used to monitor stress responses of unknown and/or known toxic chemicals using Medaka fish and may be used for the further development of biomarkers by utilizing the gene expression patterns for known contaminants.

• 한국물환경학회지
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• 제33권6호
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• pp.736-743
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• 2017

In this study, 4-nonylphenol (4-NP), an endocrine disrupting chemical, was removed by ozone treatment processes under the various experimental conditions including UV irradiation, $TiO_2$ addition. The ozone flow rate and concentration were maintained at $1.0L{\cdot}min^{-1}$ and $70{\pm}5mg{\cdot}L^{-1}$. The pH, COD and TOC of the samples were obtained every 10 minutes for 60 minutes in laboratory scale batch reactor. We found that the combination of UV irradiation and $TiO_2$ addition for ozonation improves the removal efficiency of COD and TOC in 4-NP aqueous solution. In case of the $O_3/UV/TiO_2$ system, COD and TOC were greatly reduced to 85.3 ~ 94.0% and 89.2 ~ 97.2%, respectively. 4-NP degradation rate constants, $k_{COD}$ and $k_{TOC}$, were calculated based on the COD and TOC values. Significantly, $k_{COD}$ and $k_{TOC}$ were improved in the $O_3/UV/TiO_2$ treatment process compared with single $O_3$ process, because the oxidation and the mineralization of 4-NP were increased by generating of the hydroxyl radical. The $k_{COD}$ and $k_{TOC}$ were obtained to be $5.81{\times}10^{-4}{\sim}10.8{\times}10^{-4}sec^{-1}$ and $11.9{\times}10^{-4}{\sim}19.4{\times}10^{-4}sec^{-1}$ in the $O_3/UV/TiO_2$ process. Activation energy ($E_a$) of ozone oxidation reaction based on $k_{COD}$ and $k_{TOC}$ were increased in order of $O_3/UV/TiO_2$ < $O3/UV$ < $O_3/TiO_2$ < $O_3$ process. It was confirmed that the addition of $TiO_2$ and UV irradiation to the ozone oxidation reaction significantly reduced the $E_a$ value and the degradation of 4-NP.

• 미생물학회지
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• 제52권3호
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• pp.306-312
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• 2016

미세조류는 수환경으로 유입되는 독성물질의 배출기준을 설정하거나 환경영향을 평가하기 위한 환경변화의 잠재적 생물지표이다. 본 논문에서 해양 미세조류인 녹조류 Tetraselmis suecica, 규조류 Ditylum brightwellii, 와편모조류 Prorocentrum minimum에 대한 내분비 교란물질(EDCs) 비스페놀 A (BPA)와 Aroclor 1016의 영향을 평가하였다. 처리한 EDCs에 대하여 각각의 종은 매우 다른 민감도 차이를 보였다. 각 종에 대한 50% 영향농도($EC_{50}$)는 Aroclor 1016가 BPA보다 더 유해하였다. 실험에 사용한 미세조류중에서 규조류 D. birghtwellii(0.037 mg/L BPA과 0.002 mg/L Aroclor 1016)가 다른 종보다 매우 민감하게 반응하는 것으로 조사되었다. 본 연구 결과는 수서생태계에로 배출되는 기준 농도 이상의 EDCs가 해양 생물에게 유해 효과가 있다는 것을 제시해 준다.

• 약학회지
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• 제45권6호
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• pp.694-700
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• 2001

Synthetic pyrethroids are analysis of a natural chemical moiety, pyrethrin derived from the pyrethrum plant Chrysanthemum. The natural pyrethrin structure has been modified to be highly lipophilic and photostable, creating an effective pesticide and resulting in an increased presence in the environment. Worldwide, they are commonly used insecticides against ticks, mites, mosquitoes, and as treatment for human head lice and scabies. Therefore, human exposure to their compounds in extensive. Several studies on the effects of pyrethroids on thyroid hormone regulation, estrogen and androgen function have been reported and yet little has been done try assess their potential hormonal activities. Among humans, a pyrethroid compound was suggested to be the causal agent for gynecomastia in a group of Haitian men. The reports suggest that some pyrethroid compounds are capable of disrupting endocrine function. Therefore, we examined estrogenic/antiestrogenic potential of three pyrethroid insecticides, that is permethrin, allethrin and fenvalerate in human breast cancer cell and action mechanism mediated by the estrogen receptor. Fenvalerate showed weak estrogenic activity but aallethrin and permethrin showed no effect. In combination with high levels (10$^{-10}$ M, 10$^{-11}$ M) of 17$\beta$-estradiol and three synthetic pyrethroids inhibited cert proliferations in MCF7-BUS cell by 17$\beta$-estradiol. Whereas, fenvalerate increased cell proliferative activity at lower level of estradiol (10$^{-12}$ M, 10$^{-13}$ M). The relative affinities to the estrogen receptor were observed by allethrin and permethrin treatment, but not by fenvalerate. These results indicated that some of pyrethroid insecticides may modulate estrogen functions in human breast cancer cell. The action mechanisms of estrogen receptor mediated antiestrogenicity by allethrin and permethrin were postulated.

• Environmental Analysis Health and Toxicology
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• 제20권1호
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• pp.13-21
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• 2005

The purpose of this study was to determine the effects of bisphenol A (BPA), which is known to have estrogenic activity, on the early development of medaka fish (Oryzias latipes). The fertilized eggs of medaka were treated with BPA at different concentrations for 3 weeks. Embryonic growth, deformation, hatching success, and gonadal differentiation were determined to observe the effects of this chemical. Also we tried to measure the estrogenic activity of bisphenol A using ELISA and RT-PCR methods. By using this techniques, we evaluated the induction of vitellogenin, an estrogen-regulated gene from the whole body-homogenates of larvae. At results, a reduced blood circulation was seen in embryos and peritoneal edema and hindrance of yolk-sac absorption were observed in larvae of treated group. However, BPA at the concentrations tested (2～200 ㎍/L) did not have severe adverse effects on the early life-stages. According to the observation of gonadal histology, inter-sex or sex -reversal was not found in all test fish. After the exposure was ended, vitellogenin mRNA and protein levels were measured in larvae and then their levels were found to be increased in treated group with 200㎍/L. These results indicate that BPA can induce the expression of vitellogenin in early life-stages as well as in adult male fish.

• 한국환경보건학회지
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• 제40권5호
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• pp.425-434
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• 2014

Objectives: The purpose of this study is to introduce the establishment process and results of the Seoul Metropolitan Government's road map on environmental health policy. Methods: The process consisted of expert group meetings, civic participation, research, and questionnaire survey for priority environmental health policy agenda items in Seoul. Results: The announced vision for the environmental health policy was "a healthy environment, safety in Seoul". This policy was established in order to define environment health policy initiatives for a period of five years with an aim to protect Seoul citizens' health from hazardous environmental factors. The resulting Seoul environmental health policy consisted of four areas and 16 key agenda items. The four areas were "Protection for children against hazardous materials", "Enhancement of health and safety of all", "Carcinogen-free and endocrine-disrupting chemical-free Seoul (reducing environmental exposure to hazardous materials), and "Establish the foundation of environmental health policy". Sixteen key agenda items include the enhancement of management of spaces for children, certification of environmental health status at schools, establishment of a unit responsible for the environmental health of children, strengthening environmental health management for susceptible populations (children, the aged, and the socioeconomically vulnerable), management of hazardous materials, physical hazardous factors (noise, radiation, etc.), indoor air quality, and the enhancement of monitoring, research, and regulation of environment health. Conclusion: The Seoul Metropolitan Government established an environmental health policy road map for a five-year period (2013-2017). To implement this environmental health policy, budget allocation, and detailed execution plans are required.