• Korean Journal of Ecology and Environment
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• v.40 no.3
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• pp.379-386
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• 2007

Nonylphenol (NP) has been well known as a major substance of surfactant and/or estrogenic environmental hormone. We tested toxic effects of nonylphenol on the population growth and development of aquatic organism such as algae (Microcystis aeruginosa), heterotrophic nanoflagellate (Diphylleia rotans), micro- (Brachionus calyciflorus) and macro-zooplankton (Daphnia magna) among eutrophic water food-web constituents. Dosage of NP treatment were 4 to 5 grades, according to each organism's tolerance based on pre-experiments; algae (0.01, 0.05, 0.10, 1.00 mg $L^{-1}$) Diphylleia rotans (0.5, 1,2. 5,6, 10 ${\mu}g\;L^{-1})$, Brachionus calyciflorus (0.1, 0.5, 1, 2.5, 5 ${\mu}g\;L^{-1}$), and Daphnia magna (0.5, 1, 5, 10, 50 ${\mu}g\;L^{-1}$), respectively. Toxic effects were measured by the changes of biomass of each organism after NP treatment. All experiments were triplication. As suggested, the higher concentration of NP treatment, the stronger inhibited the population growth of all organisms tested. In view of toxicity, a variety of concentration of NP showed a significant growth inhibition to organism; algae to 0.05 $mg\;L^{-1}$, D. rotans and B. calyciflorus to 1.0 ${\mu}g\;L^{-1}$, and D. magna to 5.0 ${\mu}g\;L^{-1}$, respectively. The $EC_{50}$ of each organism to the nonylphenol are as follows; 3. calyciflorus (2.49 ${\mu}g\;L^{-1}$), D. rotans (3.49 ${\mu}g\;L^{-1}$), D. magna (7.61 ${\mu}g\;L^{-1})$, and M. aeruginosa (47 ${\mu}g\;L^{-1})$. NP toxic effects on the development of zooplankton like egg production showed some differences in treatment concentration between Brachionus calyciflorus ${0.1{\sim}1NP{\mu}g\;L^{-1})$ and Daphnia magna $(0.5{\sim}5NP\;{\mu}g\;L^{-1})$. These results suggest that a strong growth inhibition of predator or grazer by the nonylphenol can stimulate the algal growth, or can play important role in evoking the nuisance algal bloom in eutrophic water with enough nutrients.

• Animal cells and systems
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• v.7 no.3
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• pp.227-235
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• 2003

Bisphenol A (BPA), nonylphenol (NP), and 4-tert-octylphenol (OP) are known endocrine disrupting chemicals (EDCs) with estrogenic activity in fish. This study compared the effects of BPA, NP and OP on in vitro vitellogenin (VTG) synthesis in primary cultures of hepatocytes of the Chinese minnow Phoxinus oxycephalus. The VTG secreted into the culture medium was measured using enzyme-linked immunosorbent assay (ELISA), which we developed in this study using an antibody prepared from homogenates of Chinese minnow egg. VTG synthesis was induced by estradiol-17$\beta$ ($E_2$) and phenols (BPA, NP and OP) treatment. $E_2$ at concentrations of 10$^{-6}$ M or higher increased VTG levels significantly (P < 0.05). Exposure to 10^5\;M\;BPA\;or\;10^-4$M NP and OPinduced in vitro VTG synthesis (P < 0.01). However, $10^-3$ M BPA, NP or OP did not induce VTG synthesis. These results suggest that SPA has the highest estrogenic potential in Chinese minnow hepatocytes. Tamoxifen, an anti-estrogen, drastically blocked the production of VTG by phenols (BPA, NP and OP) suggesting that phenols (BPA, NP and OP) may act via binding to estrogen receptor (ER) in Chinese minnow hepatocytes.

• Korean Journal of Ecology and Environment
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• v.38 no.3 s.113
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• pp.304-312
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• 2005

Effects of two endocrine disrupters (EDs) , nonylphenol (NP) and diethylhexyl phthalate (DEHP), on the population growth and morphology of two freshwater HABs (harmful algal blooms) , Microcystis aeruginosa and Stephanodiscus hantzschii, which frequently evoked the hazard bloom in an eutrophic lakes and reservoirs worldwide, were examined with seven different concentrations of EDs (0.01, 0.05, 0.1, 1,2, 2.5 and 3 ppm). Even at concentration below 0.01 ppm, NP strongly inhibited the algal growth of both M. aeruginosa and S. hantzschii, regardless of the algal growth phase. Morphologically, the algal cell treated with NP gradually lost green color in cytoplasm, became smaller in cell size, and then, was hardly seen in microscopic field. On the other hand, DEHP employed did not affect two algae at all concentrations, and rather stimulated the growth by about 10% with a treatment at 3.00 ppm compared to control. These results indicate that the continuous input of EDs, DEHP into the natural water system plays a crucial role to enhance or help an outbreak of algal bloom in eutrophic waters.

• Development and Reproduction
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• v.14 no.2
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• pp.115-121
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• 2010

The in vitro effects of nonylphenol (NP) and 2,2',4,6,6'-pentachlorobiphenyl (PCB104) on ovarian steroidogenesis of the ribbed gunnel, Dictyosoma burgeri were investigated. Oocytes taken during maturation stage were incubated with 100 ng/$m{\ell}$ of NP and PCB104 in the presence of exogenous precursor, $[^3H]-17{\alpha}$-hydroxyprogesterone ($[^3H]-17{\alpha}OHP$). Steroids were extracted from the media and the isolated oocytes, and the extracts were separated and identified by thin layer chromatography. The identities of the major metabolites were testosterone (T) and estradiol-$17{\beta}$ (E2). NP treatment inhibited production of E2 metabolite in the oocytes of 1.2, 1.3 and 1.4 mm although NP inhibited production of T metabolite at the oocytes of 1.1, 1.3 and 1.4 mm. PCB104 treatment inhibited production of T metabolite in the oocytes of all groups and E2 metabolite in the oocytes of 1.2, 1.3 and 1.4 mm. In conclusion, these results suggested that NP and PCB104 had an inhibitory effects on conversion of $[^3H]-17{\alpha}OHP$ to T and E2 during the oocyte maturation process of ribbed gunnel.

• Environmental Analysis Health and Toxicology
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• v.29
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• pp.2.1-2.7
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• 2014

Objectives Heat shock protein 90 (HSP90) is a highly conserved molecular chaperone important in the maturation of a broad spectrum of protein. In this study, an HSP90 gene was isolated from Asian paddle crab, Charybdis japonica, as a bio-indicator to monitor the marine ecosystem. Methods This work reports the responses of C. japonica HSP90 mRNA expression to cellular stress by endocrine disrupting chemicals (EDCs), such as bisphenol A (BPA) and 4-nonylphenol (NP) using real-time. reverse transcription polymerase chain reaction. Results The deduced amino acid sequence of HSP90 from C. japonica shared a high degree of homology with their homologues in other species. In a phylogenetic analysis, C. japonica HSP90 is evolutionally related with an ortholog of the other crustacean species. The expression of HSP90 gene was almost distributed in all the examined tissues of the C. japonica crab but expression levels varied among the different body parts of the crabs. We examined HSP90 mRNA expression pattern in C. japonica crabs exposed to EDCs for various exposure times. The expression of HSP90 transcripts was significantly increased in C. japonica crabs exposed to BPA and NP at different concentrations for 12, 24, 48 and 96 hours. The mRNA expression of HSP90 gene was significantly induced in a concentration- and time-dependent manner after BPA or NP exposures for 96 hours. Conclusions Taken together, expression analysis of Asian paddle crab HSP90 gene provided useful molecular information about crab responses in stress conditions and potential ways to monitor the EDCs stressors in marine environments.

• Animal cells and systems
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• v.15 no.3
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• pp.189-196
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• 2011

We investigated the in vitro effects of nonylphenol (NP) and 3,3',4,4',5-pentachlorobiphenyl (PCB126) on steroidogenesis in redlip mullet, Chelon haematocheilus, oocytes. In experiment 1, we investigated the effects of NP and PCB126 on steroid production from exogenous steroid precursors. Vitellogenic oocytes (0.75 mm in diameter) were incubated with 10 and 100 ng/ml NP or PCB126 with $[^3H]17{\alpha}$-hydroxyprogesterone as a precursor. The major metabolites produced were androstenedione, testosterone (T), estrone, and estradiol-$17{\beta}$ ($E_2$). Both NP and PCB126 increased T production and decreased $E_2$ production, except for 100 ng/ml PCB126. In experiment 2, oocytes (0.65-0.75 mm in diameter) were exposed to NP and PCB126 at different concentrations (0.01, 0.1, 1, 10, and 100 ng/mL). After the incubation, T and $E_2$ production was measured by radioimmunoassay. NP inhibited $E_2$ production at concentrations of 0.01 and 0.1 ng/ml in 0.75-mm-diameter oocytes. NP at 1 and 100 ng/mL stimulated T production, but had no observable effect on $E_2$ production. PCB126 treatment did not affect $E_2$ production at any of the concentrations tested. NP alone at 0.1 ng/mL resulted in a significant decrease in $E_2$ production in 0.65-mm-diameter oocytes. PCB126 did not show any significant effects on either T or $E_2$ production at all concentrations tested. These results suggest that NP acts like an antiestrogen at lower concentrations (0.01-0.1 ng/ml) in vitellogenic oocytes of redlip mullet.

• Proceedings of the PSK Conference
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• 2001.04a
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• pp.176.2-177
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• 2001

• Environmental Analysis Health and Toxicology
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• v.20 no.3 s.50
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• pp.195-203
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• 2005

Cytochrome P45O 17$\alpha$-hydroxylase (CYPI 7) and cytorhrome P45O aromata.ie (CYPI 9) are key steroidogenic enzymes in androgen and estrogen synthesis. ThiL study evaluated the effects of nonylphenol (NP) on CYP17 and CYP19 expression in the ovary of Sprague-Dawley rats. All female rats were administered orally with the vehicle (control, corn oil), diethylstilbestrol (DES, 5.0 $\mu$g/kg) and NP (50, 100, or 200 mg/kg/day), which was startinB when they were weaned at 21 days of age for 20 days. Twenty four hours after final dose, the animals were anelthetized with ether. Significant decreases in the uterus (wet weight) were observed with 5.0 $\mu$g/kg/day DES (78$\%$, of control) and 200 mg/kg/day NP (62$\%$ of control), respectively Additionally, ovarian weight was significantly decreased with 5.0 $\mu$g/kg/day DES (63$\%$ of control) and 200 mg/kg/day NP (72$\%$ of control). The serum estradiol levels were sligHtly lower in DES and high dose NP treatment groups, but the 74 levels were not affected by DES and NP. The expression of the ovarian CYP19 gene increased with low doses (50 and 100 mg/kg/day) of NP. while DES and high dose oi NP (200 mg/kg/day) did not affect on the CYP19 mRNA levels. In contrast to the CYP19 gene, the CYP17 gene expreLsion level was significantly down-regulated by the DES and 200 mg/ks/day NP. This result suggestE that NP inhibits ovarian estrogen synthelis by supprelsing CYP17 mRNA efprelsion, And different mechanisml might exist for the expression of Lteroidogenic CYP17 and CYP19 genes in the ovary of Sprague-Dawley rats in response to NP.

• Fisheries and Aquatic Sciences
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• v.12 no.4
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• pp.293-298
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• 2009

Several studies have reported that nonylphenol (NP) and 2,2', 4,6,6'-pentachlorobiphenyl (PCB104) exhibit estrogenic activity. To investigate the estrogenic potency of NP and PCB104 during oocyte maturation, fully vitellogenic oocytes (0.76 mm diameter in average) of yellow fin goby, Acanthogobius flavimanus, were exposed in vitro to these chemicals at different concentrations (0.1, 1, 10, 100, and 1,000 ng/mL) with the exogenous precursor $17\alpha$-hydroxyprogesterone ($17{\alpha}OHP$) 50 ng/mL in the presence or absence of human chorionic gonadotropin (HCG). The production of testosterone (T), estradiol-$17\beta$ (E2), and $17\alpha,20\beta$-dihydroxy-4-pregnen-3-one ($17\alpha20{\beta}OHP$) in response to NP or PCB104 were measured by radioimmunoassay. Steroid levels were also expressed as E2/T and E2/$17\alpha20{\beta}OHP$ ratios. In the absence of HCG, no significant differences in either NP or PCB104 treatment groups were observed. In the presence of HCG, NP treatment did not show significant differences in the production of T, E2, and $17\alpha20{\beta}OHP$ at any concentrations tested, but E2/T ratios were decreased at concentrations of 0.1, 1, 10, and 1,000 ng/mL compared with the control group. PCB104 decreased E2 production at concentrations of 0.1, 10, and 1000 ng/mL, but did not show significant differences in the production of T and $17\alpha20{\beta}OHP$ at any concentration tested. While E2/T ratios were decreased at PCB104 concentrations of 0.1, 1, 10, and 1,000 ng/mL, E2/$17\alpha20{\beta}OHP$ ratios were also decreased at 0.1, 10, and 1,000 ng/mL compared with the control. Results indicate that both NP and PCB104 appeared to have antiestrogenic effects during this phase.

• Molecular & Cellular Toxicology
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• v.5 no.1
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• pp.67-74
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• 2009

Exposures to environmental chemicals that mimic endogenous hormones are proposed for a number of adverse health effects, including infertility, abnormal prenatal and childhood development and above all cancers. In addition, recently miRNA (micro RNA) has been recognized to play an important role in various diseases and in cellular and molecular responses to toxicants. In this study, endocrine disrupting environmental toxicant, nonylphenol (NP) was treated to MCF-7 (Human breast cancer cell) and HepG2 (Human hepatocellular liver carcinoma) cell line at 3 hrs and 48 hrs time point and miRNA analysis using $mirVana^{TM}$ miRNA bioarray was performed and compared with total mRNA microarray data for the same cell line and treatment. Robust data quality was achieved through the use of dye-swap. Analysis of microarray data identifies a total of 20 and 11 miRNA expressions at 3 hrs and 48 hrs exposure to NP in MCF-7 cell line and a total of 14 and 47 miRNA expression at 3 hrs and 48 hrs exposure respectively to NP in HepG2 cell line. Expression profiling of the selected miRNA (let-7c, miR-16, miR-195, miR-200b, miR200c, miR-205, and miR-589) reveals changes in the expression of target genes related to metabolism, immune response, apoptosis, and cell differentiation. The present study can be informative and helpful to understand the role of miRNA in molecular mechanism of chemical toxicity and their influence on hormone dependent disease. Also this study may prove to be a valuable tool for screening potential estrogen mimicking pollutants in the environment.