• 한국환경보건학회지
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• 제33권3호
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• pp.207-210
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• 2007

The study was designed to determine the estrogenic effect of some penicillins on endocrine function in adult Japanese medaka (Oryzias latipes). Vitellogenin (Vtg) produced in male fish has been used for a biomarker to study endocrine disrupters. $17\beta-estradiol\;(E_2)$ was used a positive control that was induced Vtg in male fish. Result of total protein qantification and ELISA for female and male fish were exposed to $17\beta-estradiol$ 10ng/ml for $3\sim5$ days. As a result, male fish exposed to amoxicillin respectively appeared 0.75, 0.23, 8.21 and $9.36\%_{\circ}$ of 1, 10, 100 and 1000 ppm respectively, that value was elevated compared with control male fish. Male fish exposed to ampicillin respectively appeared 1.85, 4.68, 0.85 and $39.59\%_{\circ}$ of 1, 10, 100 and 1000 ppm respectively, that value was elevated compared with control male fish. This study is one of the first reports suggesting potential endocrine disruption of some penicillins in aquatic ecosystem. These results suggest that vitellogenin and estrogen receptor induction patterns alter in male medaka treated with selected estrogenic compounds, and that these results may be useful molecular biomarkers for screening estrogenic EDCs (endocrine-disrupting chemicals) in the shortest possible time.

• 약학회지
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• 제44권5호
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• pp.416-423
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• 2000

There is a growing concern that a wide variety of chemicals released into the environment can disrupt the endocrine system of fish, wildlife and humans. Endocrine disrupting chemicals (EDCs) include pesticides such as DDT lindane and atrazine, the food packaging chemicals, phthalates and bisphenol A, alkylphenol ethoxylate detergents and the chemical industry by-products, dioxins. Xenoestrogens in the environment have been argued about health risk, because of estrogen mimetic chemicals are exposed only small amounts to human. A number of in vivo and in vitro assays are now in use to assess the activity of xenoestrogens in the environment. A human breast cancer cell line (MCF-7) was used to develop in vitro screening assay for the detection of xenoestrogenic environmental pollutants. The E-SCREEN (MCF7-BUS) assay is proposed as a reliable, easy and rapid-to-perform method. To optimize and validate this method before it can be used routinely, several phenol compounds and pesticides suspected to be estrogenic were tested using I-SCREEN assay. The results showed that this method is a valuable tool for screening potential estrogen-mimicking environmental pollutants and quantitative determination of estrogeniciy.

• Asian Pacific Journal of Cancer Prevention
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• 제13권11호
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• pp.5445-5450
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• 2012

Although benign uterine leiomyomata (LMA) is the most common reproductive tumor in premenopausal women, its etiology is largely unknown. We aimed in the present study to demonstrate the potential role of environmental factors with estrogenic activity in tumor etiology by focusing on the role of aryl hydrocarbon receptor (AhR) which mediates the effects of many environmental endocrine disruptors and contributes to the loss of normal ovarian function in polluted environments. This case-control study aimed to compare the interactions between AhR and lifestyle factors in a clinical setting for the first time among 138 newly diagnosed LMA patients and 138 normal controls who lived in Tehran and Mashhad, respectively, during the last 10 years. To conduct immunohistochemical studies using appropriate monoclonal antibodies, 30 cases were selected retrospectively from 2009-2011 from the pathology departments of two university hospitals in Tehran. Although the levels of sex steroid receptors were similar in adjacent myometrium and uterine leiomyomas of all cases, AhR was significantly overexpressed (p=0.034, OR=1.667) in uterine LMA and this overexpression was correlated with living in Tehran [(p=0.04, OR=16 (1.216-210.58)], smoking[P=0.04, OR=2.085 (1.29-3.371)], living near polycyclic aromative hydrocarbon producing companies [p=0.007, OR=2.22 (1.256-3.926)] and eating grilled meat [p=0.042, OR=1.28 (1.92-3.842)]. Our study contributes to the understanding of the effects of EDCs on AhR levels as well as women's health and points out possible risk factors for the development and growth of uterine LMA. It seems that the development of LMA could be the result of interactions between hormonal and environmental factors.

• Toxicological Research
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• 제31권4호
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• pp.331-337
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• 2015

Synthetic pyrethroids (SPs) are the most common pesticides which are recently used for indoor pest control. The widespread use of SPs has resulted in the increased exposure to wild animals and humans. Recently, some SPs are suspected as endocrine disrupting chemicals (EDCs) and have been assessed for their potential estrogenicity by adopting various analyzing assays. In this study, we examined the estrogenic effects of lambda-cyhalothrin (LC) and cypermethrin (CP), the most commonly used pesticides in Korea, using BG-1 ovarian cancer cells expressing estrogen receptors (ERs). To evaluate the estrogenic activities of two SPs, LC and CP, we employed MTT assay and reverse-transcription polymerase chain reaction (RT-PCR) in LC or CP treated BG-1 ovarian cancer cells. In MTT assay, LC ($10^{-6}M$) and CP ($10^{-5}M$) significantly induced the growth of BG-1 cancer cells. LC or CP-induced cell growth was antagonized by addition of ICI 182,720 ($10^{-8}M$), an ER antagonist, suggesting that this effect appears to be mediated by an ER-dependent manner. Moreover, RT-PCR results showed that transcriptional level of cyclin D1, a cell cycle-regulating gene, was significantly up-regulated by LC and CP, while these effects were reversed by co-treatment of ICI 182,780. However, p21, a cyclin D-ckd-4 inhibitor gene, was not altered by LC or CP. Moreover, $ER{\alpha}$ expression was not significantly changed by LC and CP, while down-regulated by E2. Finally, in xenografted mouse model transplanted with human BG-1 ovarian cancer cells, E2 significantly increased the tumor volume compare to a negative control, but LC did not. Taken together, these results suggest that LC and CP may possess estrogenic potentials by stimulating the growth of BG-1 ovarian cancer cells via partially ER signaling pathway associated with cell cycle as did E2, but this estrogenic effect was not found in in vivo mouse model.

• Fisheries and Aquatic Sciences
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• 제14권1호
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• pp.22-30
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• 2011

Vitellogenin (Vg) is the precursor of vitellin (Vn), which is the major yolk protein in nearly all oviparous species, including fish, amphibians, reptiles, and most invertebrates. It is one of the most important factors during reproduction, and numerous studies have shown that Vg genes are markers of the reproductive cycle and effecter genes induced by endocrine-disrupting chemicals (EDCs). Previously, we isolated two distinct cDNAs encoding vitellogenin homologs Pj-Vg1 and Pj-Vg2 from Pandalus shrimp Pandalopsis japonica. In this study, full-length genomic sequences of Pj-Vg1 and Pj-Vg2 were determined using a PCR-based genome walking strategy. Isolated Pj-Vg1 and Pj-Vg2 genes were 11,910 and 11,850 bp long, respectively. Both Pj-Vg genes had 15 exons and 14 introns, and the splicing sites were also the same, suggesting that they arose via gene duplication. The similar structural characteristics of decapod Vg genes suggest that they are all orthologs that evolved from the same ancestral gene. Analysis of Pj-Vg1 and Pj-Vg2 expression revealed that the relative copy numbers of Pj-Vg1 and Pj-Vg2 were similar in the hepatopancreas, whereas Pj-Vg2 transcripts were also detected in the ovary. Expression of both Pj-Vg genes was induced in hepatopancreas of mature individuals, whereas only Pj-Vg2 transcripts were upregulated in the ovaries from mature animals, suggesting that both Pj-Vgs are important for oocyte development. A strong positive correlation was found between Pj-Vg1 and Pj-Vg2 transcripts in the same individual, indicating they are under the same control mechanisms. Additionally, a positive correlation was found between ovarian and hepatopancreatic Pj-Vg2 transcripts, suggesting that its dual expression is regulated by similar physiological conditions. Knowledge of the similarities and differences between the two vitellogenin-like genes, Pj-Vg1 and Pj-Vg2, would help us to understand their roles in reproduction and other physiological effects.

• Environmental Analysis Health and Toxicology
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• 제24권4호
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• pp.287-292
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• 2009

Over the a few decades, the incidences of obesity and associated metabolic syndrome diseases have been increased dramatically and resulted in a global health crisis. Recent findings suggest endocrine disrupting chemicals (EDCs) as an obesogen, because they disrupt normal development and homeostatic controls over adipogenesis and energy balance in obesity. Furthermore, risk of cardiovascular disease and mortality is elevated among those who were obese during childhood. Thus, we focused on etiology of obesity in children and performed biological monitoring of bisphenol A (BPA), which is a broadly exposed EDC in environment. Study subjects were age and sex-matched obese and normal children in Seoul (N=52; age, $8.67{\pm}1.46$ years). Exposure levels of BPA were analyzed with HPLC/FLD as a conjugated form in urine. As results, ranges of urinary BPA were 0~54.38 ${\mu}g/g$ creatinine (median, 4.57 ${\mu}g/g$ creatinine). Levels of urinary BPA were 1.7 fold higher in the obese children than those in the controls (medians of obese and control children, 7.31 and 4.25 ${\mu}g/g$ creatinine, respectively, p=0.22). In the near future, enlarge scaled studies should be performed to confirm the risk of BPA for obesity.

• 한국환경보건학회지
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• 제44권4호
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• pp.370-379
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• 2018

Objectives: Bisphenol A (BPA) has been extensively used in a variety of consumer products, resulting in widespread non-occupational human exposure. It is often detected in the human body. Studies have reported many health effects associated with endocrine and metabolic disruptions, including obesity, diabetes, hypertension, and cardiovascular diseases. This study was performed to explain the relationship between BPA exposure and obesity in the Korean adult population. Methods: The second stage of the Korean National Environmental Health Survey (KoNHES) was conducted from 2012 to 2014 with 6,478 persons participating. Using the results of the survey, we analyzed the exposure levels for BPA and the influence on obesity of BPA. Results: In model 1, the volume-based measure concentration of BPA, total, female and the 30s to 60s age group were positively related with BMI. In model 2, creatinine adjusted as a covariate and positive associations for BPA with BMI were observed in the female group and was marginally significantly associated in low body weight group. In model 3, creatinine adjusted (g/g-creatinine), BPA exposure, and BMI were positively related with sex, in females, and there was a marginally significant association with the low body weight group in the BMI categories. BMI was significantly associated with BPA in the female group in all three models. Conclusion: This study added further evidence that exposure to EDCs, include bisphenol A, is related with obesity among the general population. Given the environmental health concerns over BPA, it is necessary to develop comprehensive measures to reduce BPA exposure.

• Fisheries and Aquatic Sciences
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• 제12권4호
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• pp.276-285
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• 2009

Despite great commercial interest, relatively little has been described about molecular mechanism of bivalve reproduction. We investigated genes involved in ovarian maturation of the Yesso scallop, Patinopecten yessoensis. GSI index and histological analysis revealed that maturation of ovary begin in February and spawning period is from April to June which is similar to the previous study in the East Sea. As result of combination analysis of differential display RTPCR (DDRT-PCR) and histological examination, vitellogenin (Vg), ferritin (Ft) and ADT/ATP carrier protein (ACC) were identified as differently expressed genes in maturating ovary. Endpoint RT-PCR results showed that Vg is ovary-specific genes whereas Ft and ACC are expressed ubiquitously suggesting that Vg can be good molecular markers for ovarian development and sex determination in bivalves. Quantitative PCR results revealed that Vg were expressed highest during growth stage and appears to play a major role in oocyte maturation. On the contrary, expression of Ft was highest after spawning stage, which suggests that up-regulation may be involved in spawning and inactive stages in which the scallops recover from spawning. In addition, high level of the mitochondrial gene, ACC, may play a role in energy metabolism in maturating oocytes. Isolation and molecular studies of these key genes will expand our knowledge of the physiological changes from various exogenous factors including temperature, salinity, pH, even or numerous endocrine disrupting chemicals (EDCs) during reproductive cycle. In addition, further study of these genes implicates various industrial applications including the stable seed production, increased food quality, or economic aquaculture system.

• 환경위생공학
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• 제19권4호
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• pp.25-33
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• 2004

Acrylamide, difficult to analyze by GC and GC/MS due to the polarity and low volatility, was analyzed by LC/ESI/MS in the study. Acrylamide its(molecular weight 71amu) showed m/z=72 $(M+H)^+$ and high peak intensity at 22V in SIR mode. The mass spectrum ratios of acrylamide for qualitative identification had m/z=72 in precursor ion and m/z=55 in products ion, respectively. Those ratios at 30V in SIR mode ranged from 1: 1.4 to 1:1.17 despite various acrylamide concentrations. The ion intensity ratios of acrylamide $(m/z=72,\; [M+H]^+)$ to acrylamide isotopes $(m/z=73,\;[M+H]^+)$ ranged from 100 : 3.57 to 100 : 3.92. The results verified theoretical mass spectrum ratio that was 100:3.82. The linearity of standard calibration curve was y : 520.584x + 1815.26 with $r^2=0.99.$ In quality assurance and quality control, the recovery rate ranged from 81.64 percent to 90.97 percent and relative standard deviation was less than $10\%$ with 5 repeated injections at individual standard calibration solutions. The method was applied to analyze acrylamide in food at grocery stores. Snacks made of potatoes showed the highest acrylamide concentration followed by products made of French fries, wheat, and corn.

• 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.