• Molecular & Cellular Toxicology
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• v.1 no.1
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• pp.52-61
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• 2005

Transcript profiling is a particularly valuable tool in the field of steroid receptor biology, as these receptors are ligand-activated transcription factors and therefore exert their initial effects through altering gene expression in responsive cells. Also, an awareness of endocrine disrupting chemicals (EDCs) and their potential screening methods to identify endocrine activity have been increased. Here we developed an in-house cDNA microarray, named KISTCHIP-400 ver. 1.0, with 416 clones, based on public database and research papers. These clones contained estrogen, androgen, thyroid hormone & receptors, sex hormone signal transduction & regulation, c-fos, c-myc, ps2 gene, metabolism related genes etc. Also, to validate the KISTCHIP-400 ver. 1.0, we investigated gene expression profiles with reference hormones, $10^{8}\;M\;17{\beta}-estradiol,\;10^{-7}\;M\;testosterone\;and\;10^{-7}\;M$ progesterone in MCF-7 cell line. As the results, gene expression profiles of three reference hormones were distinguished from each other with significant and identified 33 $17{\beta}-estradiol$ responsive genes. This study is in first step of validation for KISTCHIP-400 ver. 1.0, as following step transcriptional profile analysis on not only low concentrations of EDCs but suspected EDCs using KISTCHIP-400 ver. 1.0 is processing. Our results indicate that the developed microarray may be a useful laboratory tool for screening EDCs and elucidating endocrine disrupting mechanism.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.48 no.4
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• pp.459-465
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• 2015

We tested biomarker systems [enzyme-linked immunosorbent assay (ELISA) and immunochromatography assay (ICG) kits] for the screening of endocrine-disrupting chemicals in contaminated environments using antibodies resulting from $17{\beta}$-Estradiol-induced vitellogenin (Vtg) in the wild scorpion fish Sebastiscus marmoratus. Monoclonal antibodies of two clones (S28 and S15) were used as capture and tracer antibodies for ELISA and ICG assays. ELISA detected Vtg at levels greater than $0.1{\mu}g/mL$, while ICG detected Vtg at levels greater than $1{\mu}g/mL$. However, the ICG system was able to detect antibodies from $17{\beta}$-Estradiol-induced Vtg serum that had been diluted 1,000 times. Our results suggest that previously developed biomarker assays can be used as detection systems to detect known endocrine-disrupting chemicals in contaminated environments, and to measure their activity.

• Environmental Engineering Research
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• v.21 no.2
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• pp.203-210
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• 2016

Hybrid materials were obtained modifying the bentonite (BC) and local clay (LC) using hexadecyltrimethylammonium bromide (HDTMA) or the clay were pillared with aluminum followed by modification with HDTMA. The materials were characterized by the SEM, FT-IR and XRD analytical tools. The batch reactor data implied that the uptake of $17{\beta}$-estradiol (E2) by the hybrid materials showed very high uptake at the neutral pH region. However, at higher and lower pH conditions, slightly less uptake of E2 was occurred. The uptake of E2 was insignificantly affected changing the sorptive concentration from 1.0 to 10.0 mg/L and the background electrolyte (NaCl) concentrations from 0.0001 to 0.1 mol/L. Moreover, the sorption of E2 by these hybrid materials was fairly efficient since within 30 mins of contact time, an apparent equilibrium between solid and solution was achieved, and the data was best fitted to the PSO (pseudo-second order) and FL-PSO (Fractal-like-pseudo second order) kinetic models compared to the PFO (pseudo-first order) model. The fixed-bed column results showed that relatively high breakthrough volume was obtained for the attenuation of E2 using these hybrid materials, and the loading capacity of E2 was estimated to be 75.984, 63.757, 58.965 and 49.746 mg/g for the solids BCH, BCAH, LCH and LCAH, respectively.

• The Journal of Korea CHUNA Manual Medicine
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• v.1 no.1
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• pp.125-137
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• 2000

the aim of this study was to investigate whether osteoporosis can be advanced after cerebral infarction Of not and role of female sex hormone. all animals were classified 4 groups; group of ovariectomy (OVXgroup) group of cerebral infarction( INF group), group of combination ovariectomy and cerebral infarction(OVX + INF group), naturally intact group for control data (NOR group ). cerebral infarction was made by Chen,s method with some modification and ovariectomy was performed by Wayforth,s method. experimental data was collected at 15 days, 1 month, 3 months and 6months after starting observation. serum 17 ${{\beta}-estradiol\;(E_2)}$ was determined by radioimmunoassay, for comparision of osteoporosis formation, bone density, serum osteocalcin, Serum total calcium and phosphorus, Serum AST Concentration, Serum ALT Concentration, Creatinine Concentration were also calculated. we have found that cerebral ischemia decreases not only the serum concentration of 17 ${{\beta}-estradiol\;(E_2)}$ and inhibits but also the physiologically compensatory function of the ovariectomized rats and that the decreased estrogen concentration followed by cerebral infarction have not produced osteoporosis, regretfully.

• Toxicological Research
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• v.22 no.1
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• pp.15-22
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• 2006

Many of endocrine disrupting chemicals induce effects via interaction with hormone receptors and responsive elements in target cells. We investigated endocrine disrupting effects of some food additives and contaminants including BHA, BHT, ethoxyquin, propionic acid, sorbic acid, benzoic acid, CPM, aflatoxin B1, cadmium chloride, genistein, TCDD and PCBs in yeast transformants expressing human steroid hormone receptors along with steroid responsive elements. The response limit of genetically recombinant yeast to $17{\beta}$-estradiol, testosterone and progesterone was $1{\times}10^{-16},\;1{\times}10^{-12}\;and\;1{\times}10^{-13}M$, respectively. BHT induced weak transcriptional activity in estrogen sensitive yeast, while BHA and sorbic acid interacted weakly with androgen receptor/responsive element. CPM induced transcriptional activities in all types of yeasts sensitive to steroid hormones. Zearalenone and genistein induced high transcriptional activation in estrogen sensitive yeast with relative potencies almost $10^8$ folds lower than $17{\beta}$-estradiol. TCDD induced transcriptional activation weakly in estrogen- and progesterone- sensitive yeasts. This study elucidated that recombinant yeast is a sensitive and high-throughput system and can be used for the direct assessment on chemical interactions with steroid receptors and responsive elements. Also, the present study raises the requirement of evaluation on the endocrine disrupting effects of BHT, BHA, sorbic acid, CPM and TCDD for their transcription activity in yeast screening system though weak in intensity.

• Proceedings of the Korean Environmental Sciences Society Conference
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• 2003.11a
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• pp.159-164
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• 2003

Effects of Al on vitellogenin (VTG) and VTG mRNA induction by estradiol-17 $\beta$($E_2$) were examined in primary hepatocyte culture of rainbow trout. Hepatocytes were precultured for 2 days and then E2 ($2{\times}10^{-6}$M) and Al ($10^{-6}-10^{-4}$M) were simultaneously added to the incubation medium. Hepatocytes were cultured for 5 more days. Media and hepatocytes were then analyzed by SDS-PAGE and Northern blotting for VTG and VTG mRNA, respectively. These metal had no appreciable effect on the viability of hepatocytes in culture. However, Al interfered with VTG production and VTG mRNA expression. Al reduced VTG production in a concentration-dependent way and a significant reduction accurred at Al concentrations greater than $5{\times}10^{-5}$M. VTG mRNA expression also decreased with a negative correlation with Al concentration (r＝-0.98). These results suggest that Al inhibit VTG production at the transcriptional level to reduce VTG mRNA expression.

• Korean Journal of Animal Reproduction
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• v.14 no.3
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• pp.223-230
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• 1990

This experiment was carried out to investigate in vitro maturation rate of pig follicular oocytes cultured from 30 to 48hr in TCM 199 supplemented with gonadotropins(FSH, LH) and estradiol-17$\beta$ and in vitro fertilization with ejaculated sperm preincubated in BO medium containing 2mM caffein and development of IVF oocytes. The results obtained in this experiments were as follows ; 1. In addition of hormones, in vitro maturation rate of follicular oocyte increased gradually from 36hr and 74.47% at 48hr in addition of hormones, but there was no differences among in vitro maturation rates after 36hr of culture. 2. Penetration rate of pig oocytes matured in FSH＋LH＋E2 and FSH＋E2 was 71.8%, 71.0% and significantly increased by the addition of hormones. 3. Percentage of developed oocytes was 44.4% for oocytes matured in FSH＋LH＋E2-added medium and 48.7% for oocytes matured in FSH＋E2-added medium, respectively. 4. Two to 16 cells stage embryos were obtained only when pig oocytes matuerd in vitro in hormones-added medium and 72hr after IVF. 5. From present results, it is concluded that gonadotropins and estradiol17$\beta$ can enhance in vitro fertilization and subsequent development as well as in vitro maturation pig follicular oocytes.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1993.04a
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• pp.98-98
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• 1993

어류를 이용한 독성 평가방법 및 기작에 관한 연구의 일환으로 여성 호르몬의 일종인 17$\beta$-estradiol (E$_2$)의 갑상선 종양 유발 촉진성을 검증하기 위하여 부화 후 7＋1일 된 암수동체성 어류인 점박이송사리 (Rivulu marmoratu) 를 N-methyl-N'-nitrosourea (MNU) 가 10 ppm 의 농도로 녹아있는 완충 사육수에 2시간 동안 전신 노출시켜서 갑상선 종양의 유도를 촉발 (initiation) 시킨 후, E$_2$가 포함된 시험 사료 (E$_2$ 20 mg/kg diet와 E$_2$ 80 mg/kg diet) 와 이에 대한 대조 사료를 30일간 먹여 사육하였다. 그 후 80일 동안 정상사료인 염전 새우의 유생으로 키우며 매 15일 간격으로 갑상선 종양의 발생 여부를 확인하였다. 발암원 처리 후 E$_2$ 가 함유되지 않은 사료로 사육한 대조군에서는 2.4% 의 개체에서 갑상선 종양이 유도된 반면, E$_2$ 20 mg이 함유된 사료를 투여한 실험군에서의 갑상선종양 유발율은 34%로, 대조군에서 보다 14배 정도 증가하였다 (p<0.01). MNU 처리 후 E$_2$ 60 mg/kg 사료를 먹인 실험군에서의 갑상선 종양 유발율은 E$_2$ 20 mg/kg 사료를 먹인 실험군에 비하여 발암 잠재기 (latency)가 15일 정도 단축되었다. 본 실험의 결과는 E$_2$가 다단계 (multistage)로 이루어진 갑상선 종양 발생기작에 발암 촉진제로써의 역할을 한다는 것을 입증할 뿐만 아니라, 본 종이 갑상선 종양의 발생 및 촉진기작을 연구하는데 매우 적합한 실험동물 모델임을 보여준다.

• Journal of Aquaculture
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• v.11 no.4
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• pp.557-564
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• 1998

Vitellogenin (VTG) induction in response estradiol-17${\beta}$ ($E_2$) were electrophoretically examined in primary hepatocyte cultures in rainbow trout. The hepatocytes were maintained as monolayers on positively charged dishes for up to 7 days. The viability of hepatocytes on Day 7 in cultures decreased about 20.7% and 23.6% with and without $E_2$, respecitively. The amount of DNA per dish also decreased to 13.7% and 14.0% with and without $E_2$, respectively. There were no differences in viability and DNA content between the control and $E_2$-treated culture. Moreover, the rate of VTG to total protein concentrations reached the maxium level at the addition of $10^{-6}$ M E2, to the incubation medium. However, the higher concentration of $10^{-5}$ M $E_2$ rater depressed the level.

• Environmental Mutagens and Carcinogens
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• v.20 no.1
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• pp.1-6
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• 2000

자궁내 반응에 대한 용량 의존적인 효과를 검색하기 위해 에스트로겐 agonist 및 antagonist를 사용하여 미성숙 랫드에서 uterotrophic assay를 실시하였다. 김(2000) 등은 이미 난소절제랫드를 이용한 3-day uterotrophic assay에서 에스트로겐성 물질에 대한 검색 시험을 실시한 바 있다. 양성대조물질인 17-ethinyl estradiol (EE)을 미성숙랫드에 매일 3일간 피하 투여하였으며, EE의 에스트로겐 활성을 차단하기 위한 항에스트로겐성 물질인 ZM189.154은 EE와 병행 투여하였다. 본 시험 결과 EE 1.0$\mu\textrm{g}$/kg 이상 투여군에서는 자공 비대와 fluid 저류등이 대조군에 비해 육안적으로 뚜렷하게 나타났다. 자궁무게(wet 및 blotted)는 용량 의존적으로 증가하였으며, 특히, EE 1.0$\mu\textrm{g}$/kg 이상에서는 유의성있는 증가를 나타내었다. 또한 질무게 증가도 자궁무게 증가와 일치 하였다. EE 0.3$\mu\textrm{g}$/kg을 투여한 후 ZM189, 154 0.1 mg/kg 및 1.0 mg/kg 투여와 비교할 때 EE에 의해 유도된 에스트로겐 활성이 용량 의존적으로 감소되었으나 유의성 있는 결과는 ZM189, 154 1.0 mg/kg군에서만 나타났다. 본 연구결과 EE는 uterotrophic assay에서 자궁 무게 (wet와 blotted)를 용량 의존적으로 증가시켰으며, ZM189, 154 1.0mg/kg은 EE의 에스트로겐 활성을 유의하게 감소시켰다. 따라서 미성숙 랫드를 이용한 3-day uterotrophic assay는 내분비계 장애물질 검색 시험법으로 우수하다는 것을 알 수 있었다.