• Title/Summary/Keyword: Cell Screening

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28-day Repeated-dose Toxicity Study of Flutamide, an Anti- androgenic Agent, in Rats: Establishment of Screening Methods for Endocrine Disruptors (내분비계 장애물질 검색법의 확립을 위한 항안드로젠성 물질 flutamide의 랫드 28일 반복투여 독성실험)

  • 정문구;김종춘;임광현;하창수
    • Toxicological Research
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    • v.16 no.2
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    • pp.163-172
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    • 2000
  • Recently, there is a worldwide concern that a great number of man-made chemicals have a hormone-like action both in humans and in animals. DECD is developing screening programs using validated test systems to determine whether certain substances may have an effect in humans. In the present study. the establishment oj repeated-dose toxicity test method was tried. Flutamide. an anti-androgenic agent. was administered by gavage to Sprague-Dawley rats for 28 days at dose levels of 0. 0.5. 3 and 18 mg/kg body weight (10-15 rats/sex/group) to examine the effects on general findings. especially reproductive and endocrine parameters. Clinical signs. body weights, food consumption, and sexual cycle were checked and measured. For the gross and microscopic examinations. 10 rats/sex/group were sacrificed at the end of dosing period and the remaining animals of control and high dose groups (5 each) were sacrificed after 14 days recovery. Examinations for hematology and clinical chemistry were carried out at necropsy. There were no treatment-related changes in clinical signs. body weights, food consumption. gross necropsy. hematology and clinical chemistry at all doses of both sexes. The period and regularity of sexual cycle were not adversely affected at all doses by the test agent. At 18 mg/kg. both decreased weights of prostate, seminal vesicle and epididymis in males and increased weights of spleen and thymus in females were observed. In addition, decreased number of spermatids and sperms. increased serum testosterone concentration and increased incidence (100%) of interstitial cell hyperplasia were seen in males. At 18 mg/kg of the recovery group. decreased prostate weight. reduced sperm count and increased incidence (20%) of interstitial cell hyperplasia in males and increased thymus weight in females were observed. At 3 mg/kg. reduced sperm count was found. There were no adverse effects on parameters examined at 0.5 mg/kg of both sexes. The results suggested that the potential target organs of flutamide may be accessory sexual glands including testes for males and spleen and thymus for females. Taken together. this test method was found to be a useful screening test system for endocrine disrupting chemicals.

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The Frequency and Distribution of Unexpected Red Cell Antibodies and Analysis of Antigen Exposure

  • Yang, Sung Sik;Yang, Hoi Joo;Park, Hyun Jun;Kwon, Seog Woon
    • Korean Journal of Clinical Laboratory Science
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    • v.45 no.4
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    • pp.164-169
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    • 2013
  • Red cell alloantibodies other than naturally occurring anti-A or anti-B are called unexpected red cell antibodies, and can be detected by performing an antibody screening. The frequency and distribution of unexpected antibody identified in Asan Medical Center were analyzed. We investigated a total of 135,238 cases of antibody screening test in AMC for 3 years from 2010 to 2012. Using column agglutination techniques, antibody identification tests were performed for the cases with positive antibody screening. Among 135,238 cases, 854 (0.6%) cases showed positive results of antibody screening test. In the order of frequency, 284 (33.3%) anti-Rh, 89 (10.4%) anti-MNS, 62 (7.3%) anti-Lewis, 34 (4.0%) anti-Kidd, 10 (1.2%) anti-Duffy, and 9 (1.1%) anti-P were identified. Multiple antibodies were detected in 199 (23.3%) cases. Among 381 subjects investigated for transfusion history, 299 (78.5%) had history of transfusion while 82 (21.5%) had unknown history. Thus the incidence of unexpected antibody was higher in the group with history of transfusion than the group without (p<0.001). Also, among 435 subjects investigated for the history of pregnancy, 46 (10.6%) had no history while 389 (89.4%) had history of pregnancy, showing higher incidence of unexpected antibody in the group with history of pregnancy than the group without pregnancy (p<0.001). Evaluated amounts and frequency of antigen exposure due to transfusion and pregnancy is suggested to increase the frequency of identification of unexpected antibody.

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Identification of Chinese Cabbage Sentrin as a Suppressor of Bax-Induced Cell Death in Yeast

  • Sawitri, Widhi Dyah;Slameto, Slameto;Sugiharto, Bambang;Kim, Kyung-Min
    • Journal of Microbiology and Biotechnology
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    • v.22 no.5
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    • pp.600-606
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    • 2012
  • Studies into the cell death program termed apoptosis have resulted in new information regarding how cells control and execute their own demise, including insights into the mechanism by which death-preventing factors can inhibit Bax-induced caspase activation. We investigated high temperature stress-induced cell death in Brassica rapa. Using a yeast functional screening from a Brassica rapa cDNA library, the BH5-127 EST clone encoding an apoptotic suppressor peptide was identified. However, a phylogenic tree showed that BH5-127 clusters within a clade containing SUMO-1 (Small Ubiquitin-like Modifier-1). BH5-127 was confirmed similar to have function to SUMO-1 as Fas suppression. Expression of BH5-127 showed that substantial suppression of cell death survived on SD-galactose-$Leu^-$-$Ura^-$ medium. The results suggest that BrSE ($\underline{B}$rassica rapa $\underline{S}$entrin $\underline{E}$ST, BH5-127) is one of the important regulatory proteins in programming cell death, especially in the seedling stage of Chinese cabbage.

Screening and Isolation of the Cell Differentiation Inducers from Medicinal Plants (I) (생약으로부터 세포분화유도물질의 검색 및 분리 및 분리 (I))

  • Park, Eun-Jung;Kim, Jin-Woong
    • Korean Journal of Pharmacognosy
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    • v.28 no.4
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    • pp.225-232
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    • 1997
  • 300 extracts derived from 100 plants were tested for their potential to induce HL-60 cell differentiation using NBT assay and NSE/SE staining methods. Morphological changes from suspended to adherent state of the cells were also observed by microscopic examination. In result, 55 extracts induced cell differentiation into monocyte/macrophage lineage in the NBT and the NSE assay.

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Fabrication of a Parallel Polymer Cantilever to Measure the Contractile Force of Drug-treated Cardiac Cells (약물처리된 심장세포의 세포 수축력 측정을 위한 병렬 폴리머 캔틸레버 제작)

  • Kim, Dong-Su;Lee, Dong-Weon
    • Journal of Sensor Science and Technology
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    • v.29 no.2
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    • pp.100-104
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    • 2020
  • Thus far, several in vivo biosensing platforms have been proposed to measure the mechanical contractility of cultured cardiomyocytes. However, the low sensitivity and screening rate of the developed sensors severely limit their practical applications. In addition, intensive research and development in cardiovascular disease demand a high-throughput drug-screening platform based on biomimetic engineering. To overcome the drawbacks of the current state-of-the-art methods, we propose a high-throughput drug-screening platform based on 16 functional high-sensitivity well plates. The proposed system simulates the physiological accuracy of the heart function in an in vitro environment. We fabricated 64 cantilevers using highly flexible and optically transparent silicone rubber and placed in 16 independent wells. Nanogrooves were imprinted on the surface of the cantilever to promote cell alignment and maturation. The adverse effects of the cardiovascular drugs on the cultured cardiomyocytes were systematically investigated. The 64 cantilevers demonstrated a highly reliable and reproducible mechanical contractility of the drug-treated cardiomyocytes. Real-time high-throughput screening and simultaneous evaluation of the cardiomyocyte mechanical contractility under multiple drugs verified that the proposed system could be used as an efficient drugtoxicity test platform.

Optimization and Elucidation of Interactions between Ammonium, Nitrate and Phosphate in Centella asiatica Cell Culture Using Response Surface Methodology

  • Omar Rozita;Abdullah M. A.;Hasan M. A.;Marziah M.;Mazlina M.K.Siti
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.10 no.3
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    • pp.192-197
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    • 2005
  • The effects of macronutrients $(NO_3^-,\; NH_4^+\;and\;PO_4^{3-})$ on cell growth and triterpenoids production in Centella asiatica cell suspension cultures were analyzed using the Box­Behnken response surface model experimental design. In screening and optimization experiments, $PO_4^{3-}$ as a single factor significantly influenced cell growth where increasing the phosphate level from 0.1 to 2.4 or 2.6 mM, elevated cell growth from 3.9 to $14\~16g/L$. The optimum values predicted from the response surface model are 5.05mM $NH_4^+$, 15.0mM $NO_3^-$ and 2.6mM $PO_4^{3-}$, yielding 16.0g/L cell dry weight with $99\%$ fitness to the experimental data. While the $NH_4^+-NO_3^-$ interaction influenced cell growth positively in the optimization experiment, $NH_4^+$ and $NO_3^-$ as single factors; and interactions of $NO_3^--PO_4^{3-},\;NH_4^+-PO_4^{3-}$ and $NH_4^+-NO_3^-$ were all negative in the screening experiment. Cell growth and the final pH level were positively affected by $PO_4^{3-}$, but negatively affected by $NH_4^+\;and\;NH_4^+-PO_4^{3-}$ interactions. The different effects of factors and their interactions on cell growth and final pH are influenced by a broad or narrow range of macronutrient concentrations. The productions of triterpenoids however were lower than 4mg/g cell dry weight.

Single cell heterogeneity in human pluripotent stem cells

  • Yang, Seungbok;Cho, Yoonjae;Jang, Jiwon
    • BMB Reports
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    • v.54 no.10
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    • pp.505-515
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    • 2021
  • Human pluripotent stem cells (hPSCs) include human embryonic stem cells (hESCs) derived from blastocysts and human induced pluripotent stem cells (hiPSCs) generated from somatic cell reprogramming. Due to their self-renewal ability and pluripotent differentiation potential, hPSCs serve as an excellent experimental platform for human development, disease modeling, drug screening, and cell therapy. Traditionally, hPSCs were considered to form a homogenous population. However, recent advances in single cell technologies revealed a high degree of variability between individual cells within a hPSC population. Different types of heterogeneity can arise by genetic and epigenetic abnormalities associated with long-term in vitro culture and somatic cell reprogramming. These variations initially appear in a rare population of cells. However, some cancer-related variations can confer growth advantages to the affected cells and alter cellular phenotypes, which raises significant concerns in hPSC applications. In contrast, other types of heterogeneity are related to intrinsic features of hPSCs such as asynchronous cell cycle and spatial asymmetry in cell adhesion. A growing body of evidence suggests that hPSCs exploit the intrinsic heterogeneity to produce multiple lineages during differentiation. This idea offers a new concept of pluripotency with single cell heterogeneity as an integral element. Collectively, single cell heterogeneity is Janus-faced in hPSC function and application. Harmful heterogeneity has to be minimized by improving culture conditions and screening methods. However, other heterogeneity that is integral for pluripotency can be utilized to control hPSC proliferation and differentiation.

Korean physicians' attitudes toward the prenatal screening for fetal aneuploidy and implementation of non-invasive prenatal testing with cell-free fetal DNA

  • Kim, Soo Hyun;Kim, Kun Woo;Han, You Jung;Lee, Seung Mi;Lee, Mi-Young;Shim, Jae-Yoon;Cho, Geum Joon;Lee, Joon Ho;Oh, Soo-young;Kwon, Han-Sung;Cha, Dong Hyun;Ryu, Hyun Mee
    • Journal of Genetic Medicine
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    • v.15 no.2
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    • pp.72-78
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    • 2018
  • Purpose: Physicians' attitudes may have a strong influence on women's decision regarding prenatal screening options. The aim of this study is to assess the physicians' attitudes toward prenatal screening for fetal aneuploidy including non-invasive prenatal testing (NIPT) in South Korea. Materials and Methods: Questionnaires were distributed and collected at several obstetrics-gynecological conferences and meetings. The questionnaire included 31 multiple choice and 5 fill-in-the-blank questions. Seven questions requested physicians' demographic information, 17 questions requested information about the NIPT with cell-free fetal DNA, and 12 questions requested information about general prenatal screening practices. Results: Of the 203 obstetricians that completed the survey. In contrast with professional guidelines recommending the universal offering of aneuploidy screening, only 53.7% answered that prenatal aneuploidy testing (screening and/or invasive diagnostic testing) should be offered to all pregnant women. Physicians tended to have positive attitudes toward the clinical application of NIPT as both primary and secondary screening methods for patients at high-risk for fetal trisomy. However, for patients at average-risk for fetal trisomy, physicians tended to have positive attitudes only as a secondary screening method. Physicians with more knowledge about NIPT were found to tend to inform their patients that the detection rate of NIPT is higher. Conclusion: This is the first study to investigate expert opinion on prenatal screening in South Korea. Education of physicians is essential to ensure responsible patient counseling, informed consent, and appropriate management after NIPT.

A Gap Phase-Specific Inhibitor of the Mammalian Cell Cycle from Streptomyces sp. ZF10 (Streptomyces sp. ZF-10이 생산하는 세포주기 저해제)

  • ;;Hiroyuki Osada
    • Microbiology and Biotechnology Letters
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    • v.22 no.5
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    • pp.495-498
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    • 1994
  • Genistein, a inhibitor of the progression of G$_{1}$ and G$_{2}$ phase of the mammalian cell cycle, was discovered through a unique screening system, in which effects of microbial metabolites on the cycle progression of the cultured mouse mammalian carcinoma cell were monitored by flow cytometry. The inhibitor was extracted from the fermentation broth of Streptomyces sp. ZF10 with ethyl acetate, and purified by silica gel column chromatography and HPLC.

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Antitumoral Compound, MCS-202, an Effector on Proliferation and Morphology of Human Breast Tumor Cell Line, MCF-7 (인체유암세포주 MCF-7의 형태변화와 증식에 영향을 주는 항암활성물질, MCS-202)

  • 이성우;김세은;김항섭;김환묵;이정준;김영호
    • Microbiology and Biotechnology Letters
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    • v.21 no.6
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    • pp.594-599
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    • 1993
  • In the course of screening for microbial metabolites employing human cancer cell line, we identified a mycelial extract of Streptomyces sp. 1365, which are effective on growth inhibition and morphological change of MCF-7, human breasr cancer cell line. By repeased column chromatography and recrystallization process, yellow needle crystals were obtained as an active compound and identified as resistomycin by spectral analysis.

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