• Title/Summary/Keyword: E-SCREEN assay

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Toxicogenomic Analysis and Identification of Estrogen Responsive Genes of Di (n-ethylhexyl) Phthalate in MCF-7 Cells

  • Kim, Youn-Jung;Yun, Hye-Jung;Ryu, Jae-Chun
    • Molecular & Cellular Toxicology
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    • v.1 no.3
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    • pp.149-156
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    • 2005
  • Di (n-ethylhexyl) phthalate (DEHP) is thought to mimic estrogens in their action, and are called endocrine disrupting chemicals. DEHP is used in numerous consumer products, especially those made of flexible polyvinyl chloride and have been reported to be weakly estrogenic. In this study, DEHP were tested for estrogenic properties in vitro models and with microarray analysis. First, the E-screen assay was used to measure the proliferation of DEHP in MCF-7 cells, a human breast cancer cell line. DEHP induced an increase in MCF-7 cell proliferation at concentration of $10^{-4}M$. Second, we carried out a microarray analysis of MCF-7 cells treated with DEHP using human c-DNA microarray including 401 endocrine system related genes. Of the genes analyzed, 60 genes were identified showing significant changes in gene expression resulting from DEHP. Especially, 4 genes were repressed and 4 genes were induced by DEHP compared to $17{\beta}-estradiol$. Among these genes, trefoil factor 3 (intestinal), breast cancer 1, early onset and CYP1B1 are involved in estrogen metabolism and regulation. Therefore it suggests that these genes may be associated with estrogenic effect of the DEHP on transcriptional level. The rationale is that, as gene expression is a sensitive endpoint, alterations of these genes may act as useful biomarkers to define more precisely the nature and level of exposure to kinds of phthalates.

Evaluation of Endocrine Disrupting Chemicals-Complex Mixture in Diesel Exhaust Respirable Particulate Matter

  • Ryu, Byung-Tak;Jang, Hyoung-Seok;Kim, Yun-Hee;Kim, Soung-Ho;Lee, Do-Han;Han, Kyu-Tae;Oh, Seung-Min;Chung, Kyu-Hyuck
    • Proceedings of the Korea Society of Environmental Toocicology Conference
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    • 2003.05a
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    • pp.195-195
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    • 2003
  • It is well known that diesel exhaust particulate matter contains mutagenic PAHs, such as benzo[${\alpha}$]pyrene, benz[${\alpha}$]anthracene, chrysene, etc. Therefore it is suspected that these chemicals act on estrogen receptor and reveal endocrine-disrupting effects. Recent attention has focused on causative chemicals of endocrine-disrupting effects. We examined the estrogenic activity of respirable diesel exhaust particulate matter derived from diesel powered vehicle. PM2.5 diesel exhaust of vehicle was collected using a high volume sampler equipped with a cascade impactor. Diesel exhaust samples were fractionated according to EPA methods. The presence of estrogenic and antiestrogenic chemicals in PM 2.5 diesel exhaust was determined using E-screen assay. To quantitatively assess the estrogenic and antiestrogenic activities in diesel exhaust particulate matter, estradiol equivalent concentration (bio-EEQ) was calculated by comparing the concentration response curve of the sample with those of the estrogen calibration curve. Weak estrogenic activities and strong antiestrogenic activities were detected in the crude extract and moderately polar fractions. Higher antiestrogenic potency was observed with higher EROD activities in aliphatic and aromatic compounds fraction. In conclusion, estrogenic/antiestrogenic-like activities were present in diesel exhaust particulate matter. However, the health consequences of this observation was unknown, the presence of these activities may contribute to and exacerbate adverse health effect evoked by diesel exhaust particulate matter.

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Aberrant Expression of CCAT1 Regulated by c-Myc Predicts the Prognosis of Hepatocellular Carcinoma

  • Zhu, Hua-Qiang;Zhou, Xu;Chang, Hong;Li, Hong-Guang;Liu, Fang-Feng;Ma, Chao-Qun;Lu, Jun
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.13
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    • pp.5181-5185
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    • 2015
  • Background: CCAT1 has been reported to be linked with pathogenesis of malignancies including colon cancer and gastric cancer. However, the regulatory effect of CCAT1 in hepatocellular carcinoma (HCC) remains unclear. The purpose of this research was to identify any role of CCAT1 in the progression of HCC. Materials and Methods: Real time-PCR was performed to test the relative expression of CCAT1 in HCC tissues. A computation screen of CCAT1 promoter was conducted to search for transcription-factor-binding sites. The association of c-Myc with CCAT1 promoter in vivo was tested by Pearson correlation analysis and chromatin immunoprecipitation assay. Additionally, Kaplan-Meier analysis and Cox proportional hazards analyses were performed. Results: c-Myc directly binds to the E-box element in the promoter region of CCAT, and when ectopically expressed increases promoter activity and expression of CCAT1. Moreover, Kaplan-Meier analysis demonstrated that the patients with low expression of CCAT1 demonstrated better overall and relapse-free survival compared with the high expression group. Cox proportional hazards analyses showed that CCAT1 expression was an independent prognostic factor for HCC patients. Conclusions: The findings demonstrated CCAT1, acting as a potential biomarker in predicting the prognosis of HCC, is regulated by c-Myc.

Antibiotics (항균제)

  • 박승함
    • Korean Journal of Microbiology
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    • v.9 no.2
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    • pp.86-93
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    • 1971
  • Escherichiae-like organisms were isolated from rectal specimens of 56 children who were either in preschool age or in elementary school. The isolated strains were subjected to tests to screen enteropathogens producing heat-labile enterotoxin and susceptibility test to various antibiotics by disc diffusion method on agar plates. Production of heat-labile enterotoxin by the strains was assyed in the sensitive and reproducible cultured adrenal tumor cell system. The assay was sterodogenesis of the cell in the presence of heat-labile enterotoxin. Among 56 strains, gave positive reaction in the test of toxin production. This meant that about 10% of the children population objected to the study harbored the toxigenic strain of enteropathogenes. Some of these toxigenic strains were resistant to the antibiotics employed in the test. This study suggested that considerable population in Korea may harbor entertoxigenic E. coli as a part of intestinal normal flora. The toxigenic strains which are resistant to antibiotics may bring issue of public health in the future.

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Estrogenic Activity of Sanguiin H-6 through Activation of Estrogen Receptor α Coactivator-binding Site

  • Trinh, Tuy An;Park, Eun-Ji;Lee, Dahae;Song, Ji Hoon;Lee, Hye Lim;Kim, Ki Hyun;Kim, Younghoon;Jung, Kiwon;Kang, Ki Sung;Yoo, Jeong-Eun
    • Natural Product Sciences
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    • v.25 no.1
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    • pp.28-33
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    • 2019
  • A popular approach for the study of estrogen receptor ${\alpha}$ inhibition is to investigate the protein-protein interaction between the estrogen receptor (ER) and the coactivator surface. In our study, we investigated phytochemicals from Rubus coreanus that were able to disrupt $ER{\alpha}$ and coactivator interaction with an $ER{\alpha}$ antagonist. The E-screen assay and molecular docking analysis were performed to evaluate the effects of the estrogenic activity of R. coreanus extract and its constituents on the MCF-7 human breast cancer cell line. At $100{\mu}g/mL$, R. coreanus extract significantly stimulated cell proliferation ($574.57{\pm}8.56%$). Sanguiin H6, which was isolated from R. coreanus, demonstrated the strongest affinity for the $ER{\alpha}$ coactivator-binding site in molecular docking analysis, with a binding energy of -250.149. The initial results of the study indicated that sanguiin H6 contributed to the estrogenic activity of R. coreanus through the activation of the $ER{\alpha}$ coactivator-binding site.

Development of reverse transcription loop-mediated isothermal amplification assays for point-of-care testing of avian influenza virus subtype H5 and H9

  • Zhang, Songzi;Shin, Juyoun;Shin, Sun;Chung, Yeun-Jun
    • Genomics & Informatics
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    • v.18 no.4
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    • pp.40.1-40.8
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    • 2020
  • Avian influenza (AIV) outbreaks can induce fatal human pulmonary infections in addition to economic losses to the poultry industry. In this study, we aimed to develop a rapid and sensitive point-of-care AIV test using loop-mediated isothermal amplification (LAMP) technology. We designed three sets of reverse transcription LAMP (RT-LAMP) primers targeting the matrix (M) and hemagglutinin (HA) genes of the H5 and H9 subtypes. RT-LAMP targeting the universal M gene was designed to screen for the presence of AIV and RT-LAMP assays targeting H5-HA and H9-HA were designed to discriminate between the H5 and H9 subtypes. All three RT-LAMP assays showed specific amplification results without nonspecific reactions. In terms of sensitivity, the detection limits of our RT-LAMP assays were 100 to 1,000 RNA copies per reaction, which were 10 times more sensitive than the detection limits of the reference reverse-transcription polymerase chain reaction (RT-PCR) (1,000 to 10,000 RNA copies per reaction). The reaction time of our RT-LAMP assays was less than 30 min, which was approximately four times quicker than that of conventional RT-PCR. Altogether, these assays successfully detected the existence of AIV and discriminated between the H5 or H9 subtypes with higher sensitivity and less time than the conventional RT-PCR assay.

Evaluation of Gestational Age Median Value by Use of the Quad Test with Dimeric Inhibin A for Korea Pregnant Women (Inhibin-A를 추가한 Quad Test의 한국인 산모의 임신주수별 Median치에 대한 평가)

  • Yoo, Ja-Young;Choi, Sam-Kyu;Cho, Young-Suk;Hwang, Do-Young
    • Korean Journal of Clinical Laboratory Science
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    • v.37 no.1
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    • pp.56-60
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    • 2005
  • Human chorionic gonadotrophin (hCG) and unconjugated estriol (uE3) were added to AFP to make what is commonly known as the Triple test. The Triple test combines results from these three tests and has been a standard screening procedure for several years. Recent studies have demonstrated the usefulness of adding inhibin-A to Down's syndrome risk assessment. The Quad test adds dimeric Inhibin-A (DIA) to the three other markers and uses the same computer program to calculate risk factors. Testing was performed between 14 and 21 weeks of gestation. Sample size were 648 samples and period of study was from 1, July, 2004 to 30, September, 2004. Used analytical methods for AFP, hCG and uE3 were radioimmunoassay (RIA) and dimeric inhibin A was enzyme-linked immunosorbent assay (ELISA). Adding dimeric inhibin-A as a fourth marker to the standard triple test increases the detection rate from 62 % to 75 % with a false-positive rate of 5%. The DIA based Quad test has been shown to be the most effective second trimester screening test for Down's syndrome suitable for routine use. Increased DIA values are observed during normal pregnancy where a bimodal pattern response is seen. Values increase during the first trimester, decline after 14 weeks, and re-ascend between 17-25 weeks. Values for DIA may be additionally elevated during a Down's syndrome pregnancy. Dimeric inhibin A is a glycoprotein hormone made by the ovary and placenta. DIA levels are twice as high in Down's syndrome pregnancies. AFP, hCG, and uE3 levels vary with gestational age, and incorrect gestational dating will influence results. DIA levels do not vary substantially with gestational age, resulting in greater screening accuracy. Although the Quad test is an improvement over the Triple test, it is important to underscore the fact that a positive test on both should be done. Most women who initially screen positive will be found to be carrying normal babies when amniocentesis and definitive diagnostic chromosome analysis are done.

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Representative Parameter of Immunostimulatory Ginseng Polysaccharide to Predict Radioprotection (방사선 방어효과 예측 가능한 면역증강 인삼 다당체의 활성인자)

  • Son, Hyeog-Jin;Shim, Ji-Young;Ahn, Ji-Yeon;Yun, Yeon-Sook;Song, Jie-Young
    • Journal of Radiation Protection and Research
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    • v.33 no.3
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    • pp.99-104
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    • 2008
  • According to the increase in the use of radiotherapy to cancer patients, many approaches have been tried to develop new agents for the protection of surrounding normal tissues. However, it is still few applied in the clinic as a radioprotector. We aim to find a representative parameter for radioprotection to easily predict the activity of in vivo experiment from the results of in vitro screening. The polysaccharide extracted from Panax ginseng was used in this study because the immunostimulator has been regarded as one of the radioprotective agent category and was already reported having a promising radioprotective activity through the increase of hematopoietic cells and the production of several cytokines. Mitogenic activity, AK cells activity and nitric oxide production were monitored for the in vitro immunological assay, and endogenous colony-forming unit (e-CFU) was measured as in vivo radioprotective parameter. The immunological activity was increased by the galactose contents of ginseng polysaccharide dependently. The result of this study suggests that mitogenic activity of splenocytes demonstrated a good correlation with in vivo radioprotective effect, and may be used as a representative parameter to screen the candidates for radioprotector.

Analysis of Protein Domain for Interaction between α-COP and ε-COP in Aspergillus nidulans (Aspergillus nidulans 분비소낭 구성요소인 α-COP과 ε-COP의 결합 부위 분석)

  • Song, Eun-Jung;Kim, Ki-Hyun;Lee, Hwan-Hee;Park, Jeong-Seok;Kang, Eun-Hye;Park, Hee-Moon
    • The Korean Journal of Mycology
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    • v.40 no.4
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    • pp.224-228
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    • 2012
  • In order to screen interactor(s) of the Aspergillus nidulans ${\alpha}$-COP of COPI vesicle, we performed the yeast two hybrid screening by using the gene for A. nidulans ${\alpha}$-COP as a bait and identified ${\varepsilon}$-COP of the COPI vesicle as an interacting protein. The A. nidualns gene for the ${\varepsilon}$-COP was designated $aneA^+$ ($\underline{A.}$ $\underline{n}$idulans $\underline{e}$psi-lone-COP), which encoded 296 amino acid residues with high level of identity with orthologs from other fungi. Domain analyses with yeast two-hybrid system suggested that the interaction between ${\alpha}$-COP and ${\varepsilon}$-COP relied on the C-terminus of both proteins, and that the N-terminal WD domian of ${\alpha}$-COP and the TPR region of ${\varepsilon}$-COP were not essential but required for the enhancement of the interaction. These results indicate that the interaction mode between ${\alpha}$-COP and ${\varepsilon}$-COP of COPI vesicle is evolutionarily well conserved in eukaryotes.

Antioxidant Activity and Physiological Function of the Anomala albopilosa Extracts (청동풍탱이(Anomala albopilosa)추출물의 항산화성 및 생리기능)

  • Yoon, Weon-Jong;Lee, Jung-A;Kim, Ji-Young;Kim, Sang-Bum;Park, Soo-Yeong
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.36 no.6
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    • pp.670-677
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    • 2007
  • We analyzed antioxidant and physiological activities to investigate the functional effects of ethanol extracts of Anomala albopilosa imago and A. albopilosa larva. In order to effectively screen for anti-inflammatory agents, we first investigated the antioxidant activities such as DPPH radical scavenging capacity, superoxide radical scavenging capacity, xanthine oxidase inhibitory activity, and nitric oxide scavenging capacity of the A. albopilosa extracts. By the screening system, we found that A. albopilosa extracts had antioxidant activity which increased with increments of the extract concentration. Moreover, we examined the inhibitory effect of the A. albopilosa extracts on the production of anti-inflammatory factors that the nitric oxide (NO), inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2) and prostaglandin $E_2\;(PGE_2)$ production activated with LPS ($1{\mu}g/mL$) in murine macrophage cell line RAW 264.7. A. albopilosa extracts potentially inhibited the iNOS and COX-2 in a dose-dependent manner. The inhibition of iNOS activity was correlated with the decrease in nitrite levels. Additionally, the $PEG_2$ production is markedly inhibited after a treatment with the A. albopilosa extracts.