• Title/Summary/Keyword: tcdd

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Development of Luciferase Reporter Gene-based Cell Bioassay for the Aromatic Hydrocarbon Receptor Agonists

  • Kim, Sun-Young;Choi, Eun-Jung;Yang, Jae-Ho
    • The Korean Journal of Physiology and Pharmacology
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    • v.10 no.6
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    • pp.349-354
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    • 2006
  • The aromatic hydrocarbon receptor (AhR) is a ligand-activated transcription factor that mediates many of the biological and toxicological effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD, dioxin) and related chemicals. The application of recombinant reporter plasmid such as the firefly luciferase gene has proven to be a very effective method to detect these chemicals. The bioassay system, CALUX, is sensitive in directly detecting AhR-agonists from a variety of environmental and biologic materials. However, responses of the AhR-dependent bioassays are dependent on the cell types used. Thus, we developed a sensitive bioassay using the recombinant mouse hepatoma cell (Hepa1c1c7) for the determination of dioxins. The recombinant cell line was stably transfected with firefly luciferase reporter gene (pGudLuc1.1). The transfected cells showed the highest induction of luciferase activity at 4.5 hr and a decrease beyond this time point. The system showed the highest sensitivity of detection ever reported. Upon TCDD exposure cells showed 2 fold increase at 10 pM and 7 fold increase at 100 pM, respectively. The passage number after the transfection played an important role in the sensitivity. The increase of passage number tended to increase the sensitivity of the cells up to 15. The media without phenol red showed a higher induction rate than with phenol red, suggesting the preferable use of phenol red-free media for the bioassay. Since each of the assays has unique characteristics that make them suitable for some screening applications and not others, development of sensitive bioanalytical methods based on a variety of cellular systems in a key to the successful determination of dioxins. The bioassay system developed in this study will contribute to further development of successful screening the AhR agonists among the environmental mixture. In addition, the rapid and sensitive nature of this cellular system can be applied as a valuable tool to screen the dioxin-like moieties among the prodrugs at the initial stage, thereby expediting the new drug discovery.

Multiphase Modeling on the Convective Transport of an Organic Solvent through Unsaturated Soils (비포화 토양층 내 유기 용매의 이류 이동에 대한 다상 모델링)

  • Lee Kun Sang
    • Journal of Soil and Groundwater Environment
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    • v.9 no.3
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    • pp.20-26
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    • 2004
  • In-situ photolysis is one of the most promising ways to clean up a soil contaminated with 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). This study focuses on the mathematical description and model development of the convective upward transport of an organic solvent driven by evaporation and photodecomposition at the surface as the major transport mechanism in the clean up process. A finite-element-based numerical model was proposed to incorporate effects of multiphase flow on the distribution of each fluid, gravity as a driving force, and the use of van Genutchen equation for more accurate description of k-S-p relations. This paper presents results of extensive numerical calculations conducted to investigate the various parameters that play a role in the solvent migration through a laboratory-scale unsaturated soil column. The numerical results indicate that gravity affects significantly on the fluids distribution and evaporation for highly permeable soils. The soil texture has a profound influence on the fluid saturation profile during evaporation process. The amount of solvent convective motion increases with increasing evaporation rates and decreasing initial water saturation. Simulations conducted in this study have shown that the developed model is very useful in analyzing the effects of various parameters on the convective migration of an organic solvent in the soil environments.

Histone deacetylation effects of the CYP1A1 promoter activity, proliferation and apoptosis of cells in hepatic, prostate and breast cancer cells

  • K. N. Min;K. E. Joung;M. J. Cho;J. Y. An;Kim, D. K.;Y. Y. Sheen
    • Proceedings of the Korea Society of Environmental Toocicology Conference
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    • 2003.10a
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    • pp.181-181
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    • 2003
  • We have studied the mechanism of action of TCDD on CYP1A1 promoter activity in both Hepa Ⅰ and MCF-7 cells using transient transfection system with p1A1-Luc reporter gene. When HDAC inhibitors, such as trichostatin A, HC toxin and a novel HDAC inhibitor, IN2001 were cotreated with TCDD to the cells transfected with plAt-Luc reporter gene, the basal promoter activity of CYP1A1 was increased by HBAC inhibitors. Also, in MCF-7 human breast cancer cells, HDAC inhibitors, such as IN2001 and trichostatin A increased the basal activity of CYP1A1 promoter but TCDD stimulated CYP1A1 promoter activity was not changed by HDAC inhibitors. And, in stably-transfected Hepa Ⅰ cells with p1A1-Luc, HDAC inhibitors increased the basal promoter activity only Also, we have investigated the effects of HDAC inhibitors on the human breast and prostate cancer cells in terms of cell proliferation and apoptosis based on SRB assay. IN2001 as well as trichostatin A inhibited the MCF-7, MDA-MB-231, MDA-MB-468, T47D, ZR75-1, PC3 cell growth dose-dependently. The growth inhibition of these cells with HDAC inhibitors was associated with profound morphological change, which suggests the HDAC inhibitors induced apoptosis of cells. The result of cell cycle analysis after 24h exposure of IN2001 showed G2/M cell cycle arrest in MCF-7 cells and apoptosis in T47D and MDA-MB-231 cells.

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Prediction of Adsorption Isotherms and Diffusivity on Activated Carbon for Persistent Organic Pollutant(2,3,7,8-TCDD) (활성탄 위에서 잔류성 유기 오염물질(2,3,7,8-TCDD)의 등온 흡착식 및 확산계수 예측)

  • Lim, Young-Il;Son, Hae-Jeong;Lee, Ohsung;Nam, Kyong-Soo;Yoo, Kyoung-Seun
    • Korean Chemical Engineering Research
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    • v.47 no.6
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    • pp.747-754
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    • 2009
  • In this study, adsorption isotherms of o-DCB(ortho-dichlorobenzene) on an activated carbon heated at $1000^{\circ}C$ for 24 hours were obtained by experiment and were predicted by using molecular simulation. The initial molecular structure of the activated carbon was designed on the basis of its molecular formula and functional groups ratio measured experimentally. Then, the molecular structure was optimized using the COMPASS(condensed-phase optimized molecular potentials for atomistic simulation studies) force field. The particle porosity, specific surface area, and particle density obtained from the optimized molecular structure of activated carbon were compared with those experimental data. The errors between experimental data and simulation results of the particle porosity, specific surface area, and particle density were shown as 7.6, 3.8, and 2.8%, respectively. Adsorption isotherms constants of o-DCB are calculated by the GCMC(grand canonical Monte Carlo) method in the optimized molecular structure of activated carbon. The simulation result of the adsorption isotherms showed an error of under 3%, compared to that of experimental data. Adsorption isotherms, adsorption heat and pore diffusivity of 2,3,7,8-TCDD(tetrachlorodibenzo-p-dioxin) was finally obtained in the same molecular structure of the activated carbon as used for o-DCB. Thus, adsorption characteristics of persistent organic pollutants on activated carbon, which are not easy to experimentally evaluate, are predicted by the molecular simulation.

Serum Concentration and Exposure History of Dioxins and Organochlorine Pesticides among Residents around the Camp Carroll Area (캠프캐럴 인근 주민에서 다이옥신류 및 유기염소계 농약의 혈중 농도 및 노출력)

  • Bae, Sang Geun;Kim, Geun-Bae;Cho, Yong-Sung;Lee, Yu-mi;Lee, Duk Hee;Yang, Wonho;Ju, Young-Su;Lee, Kwan;Min, Young-Sun;Lim, Hyun-Sul
    • Journal of Korean Society of Occupational and Environmental Hygiene
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    • v.26 no.3
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    • pp.277-285
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    • 2016
  • Objectives: This study was performed in order to evaluate whether 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) could be detected among residents living near Camp Caroll in Waegwan and whether serum concentrations of dioxins, including 2,3,7,8-TCDD, and organochlorine pesticides (OCPs) are associated with length of residence. Methods: Study subjects totaled 113 (for dioxins) and 190 (for OCPs) adults who were selected from participants in a medical investigation. Serum concentrations of dioxins and OCPs were measured using HRGC/HRMS. Information on length of residence was obtained through questionnaires. Results: 2,3,7,8-TCDD was not detected in serum among all subjects. When length of residence was classified as a categorical variable, after adjusting for confounding variables, only residents living in Waegwan for 40 years or longer tended to have high total TEQ values and 2,3,4,7,8-PeCDF with marginal significances. There was no dose-response relation between length of residence and serum concentrations of these chemicals. In multiple regression models with continuous values of the length of residence, total TEQ value and 1,2,3,4,6,7,8-HpCDF were positively associated with length of residence. However, they explained about 3-5% of total variations of serum concentrations of these compounds, while age, consumption of fatty fish, body mass index, alcohol drinking, and cigarette smoking were main variables affecting serum concentrations of dioxins or OCPs. Conclusions: In the current study, high concentrations of certain compounds were mainly observed among persons who lived in Waegwan for at least for 40 years without a dose-response relation. Therefore, it seems difficult to conclude that length of residence meaningfully contributed to the current serum concentrations of dioxins or OCPs among residents in Waegwan. However, considering the half-life of 2,3,7,8-TCDD and indirect exposure routes, the limitations of the current study design should be considered in the interpretation of the study findings.

A Study on the Correlation between Categorization of the Individual Exposure Levels to Agent Orange and Serum Dioxin Levels Among the Korean Vietnam Veterans (베트남 참전 제대 군인의 범주화된 에이전트 오렌지 개인 폭로량과 혈청 다이옥신 측정치와의 상관성에 관한 연구)

  • Kang, Han-K.;Lim, Hyun-Sul;Cheong, Hae-Kwan;Lim, Min-Kyung;Kim, Joung-Soon
    • Journal of Preventive Medicine and Public Health
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    • v.34 no.1
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    • pp.80-88
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    • 2001
  • Objectives : In an epidemiologic study on the health impact of Agent Orange exposure, the valid estimation of exposure level is the most important step. Based on recent studies, we examined the correlation between exposure levels categorized by personal exposure estimates and serum 2,3,7,8-tetrachlorodibenzo-p-dioxin (2,3,7,8-TCDD, Dioxin), exploring the possibility of utilizing the exposure level as a surrogate for the estimate of exposure to agent orange. Methods : During the study period (Jan 1996-Feb 1996), blood specimens of 745 subjects taken randomly among 1,329 persons and kept frozen, were analyzed for 2,3,7,8-TCDO and six other dioxin congeners. The serum dioxin and congeners were measured in 1998 by CDC, adjusted for serum lipids. We categorized the total exposure scores into five groups based on Agent Orange exposure data collected by interview and military records. Pearson and Spearman's correlation coefficients & multiple regression analysis were used to identify the relationship of the exposure level categorized with serum concentration of 2,3,7,8-TCDD, and six other dioxin congeners. Results : Dioxin and the other congeners, except 1,2,4,6,7,8-HpCDD, showed significant correlations to exposure categories (p<0.005): 2,3,7,8-TCDD and OCDD showed positive correlations, whereas the other congeners did negative. The values of 2,3,7,8-TCDD differed according to exposure category and proportionally increased from the low exposure group to the high, a dose-response relationship, even after other possible confounding variables were adjusted for. In multiple regression analysis, age$(\beta=0.033)$, dioxin$(\beta=0.433)$, 1,2,3,7,8-PeCDD$(\beta=-0.998)$, 1,2,3,4,7,8-HxCDD$(\beta=-0.773)$, 1,2,3,6,7,8-HxCDD$(\beta=0.255)$, 1,2,3,7,8,9-HxCDD$(\beta=-3.468)$, 1,2,3,4,6,7,8-HpCDD$(\beta=0.109)$ we re found to be significantly related to the total exposure score(p<0.005). Conclusion : This study demonstrated that the use of such categorizations as a surrogate measure of agent orange exposure in identifying exposure degrees in a health impact study is valid.

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Regulation of CYP1A1 and Inflammatory Cytokine by NCOA7 Isoform 4 in Response to Dioxin Induced Airway Inflammation

  • Cho, Sung-Hwan;Park, Shin Young;Lee, Eun Jeong;Cho, Yo Han;Park, Hyun Sun;Hong, Seok-Ho;Kim, Woo Jin
    • Tuberculosis and Respiratory Diseases
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    • v.78 no.2
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    • pp.99-105
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    • 2015
  • Background: Aryl hydrocarbon receptor (AhR), a ligand-dependent transcription factor, binds to a wide variety of synthetic and naturally occurring compounds. AhR is involved in the regulation of inflammatory response during acute and chronic respiratory diseases. We investigated whether nuclear receptor coactivator 7 (NCOA7) could regulate transcriptional levels of AhR target genes and inflammatory cytokines in 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD)-treated human bronchial epithelial cells. This study was based on our previous study that NCOA7 was differentially expressed between normal and chronic obstructive pulmonary disease lung tissues. Methods: BEAS-2B and A549 cells grown under serum-free conditions were treated with or without TCDD (0.15 nM and 6.5 nM) for 24 hours after transfection of pCMV-NCOA7 isoform 4. Expression levels of cytochrome P4501A1 (CYP1A1), IL-6, and IL-8 were measured by quantitative real-time polymerase chain reaction. Results: The transcriptional activities of CYP1A1 and inflammatory cytokines were strongly induced by TCDD treatment in both BEAS-2B and A549 cell lines. The NCOA7 isoform 4 oppositely regulated the transcriptional activities of CYP1A1 and inflammatory cytokines between BEAS-2B and A549 cell lines. Conclusion: Our results suggest that NCOA7 could act as a regulator in the TCDD-AhR signaling pathway with dual roles in normal and abnormal physiological conditions.