• Title/Summary/Keyword: in vitro detection system

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Verification of Estrogenic Activities in Ethanol Extracts of Oriental Herbal Medicines using In vitro Detection System (In vitro 검출시스템을 이용한 한약재 추출물로부터의 에스트로겐 활성의 검증)

  • Lee Sang Hyeon
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.17 no.4
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    • pp.1054-1058
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    • 2003
  • In order to evaluate the direct effect of estrogenic compounds in oriental herbal medicines, the estrogenic activity was measured using an in vitro detection system. For this system, human breast cancer cell line MCF7 was transfected using an estrogen responsive CAT reporter plasmid. Estrogenic activities of Platycodi radix, Astragali radix and Glycyrrhizae radix were evaluated using this system. Estrogenic activity of a 500 ㎍/ml ethanol extract of Platycodi radix was as same as that of a 10/sup -8/ M standard solution (17β-estradiol) and activity of a 50 ㎍/ml ethanol extract was between those of a 10/sup -8/ M and a 10/sup -9/ M standard solutions. In addition, estrogenic activity of a 50 ㎍/ml ethanol extract of Platycodi radix was as same as that of a 10/sup -10/ M standard solution. The same activity patterns were observed in the system which was treated by Astragali radix or Glycyrrhizae radix extracts. The most effective activity was observed in a system which was treated by Platycodi radix extract, but the least activity was observed by Glycyrrhizae radix extract. In this result, it was confirmed that Platycodi radix, Astragali radix and Glycyrrhizae radix extracts possess estrogenic compounds.

Establishment of an In Vitro TCD (Testosterone Compound Detection) System (테스토스테론 물질 검출을 위한 in vitro TCD 시스템 구축)

  • Lee, Dong-Geun;Jo, Jung-Kwon;Lee, Sang-Hyeon
    • Journal of Life Science
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    • v.29 no.10
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    • pp.1159-1163
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    • 2019
  • Although there is a growing interest in male menopause, a phenomenon associated with male hormone depletion, current kits using antibodies to quantify male hormones are expensive. In this study, we constructed an in vitro system for verifying the activity or concentration of male steroid hormones using a transcriptional activity test. A reporter plasmid, pGL2-Neo-ARE-AdE1BTATA, which reacts to testosterone, was constructed. In this plasmid, the ARE-AdE1bTATA sequences can be bounded by the testosterone - androgen receptor complex to express luciferase as a reporter. Then, a stable transfection was performed on the human prostate cancer cell line, LNcap-LN3. The constructed LNcap-LN3/pGL2-Neo-ARE-AdE1BTATA testosterone compound detection (TCD) system showed quantitatively proportional luciferase activities to concentrations of $10^{-13}$ to $10^{-8}M$ of standard testosterone. The established in vitro TCD system will contribute to the development of materials for health/functional foods and drugs as it will be possible to search en masse for testosterone-like or testosterone-inhibiting substances derived from natural materials.

Verification of Estrogenic Activity in Ethanol Extracts of Marine Organisms Using in vitro Test System. (In vitro 검출시스템을 이용한 해양생물 추출물로부터 에스트로겐 활성 검증)

  • 하종명;이상현
    • Journal of Life Science
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    • v.13 no.6
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    • pp.799-804
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    • 2003
  • In order to verify the occurrence of an estrogenic compound in natural products, the estrogenic activity was measured using an in vitro detection system. For this system, human breast cancer cell line MCF7 was transfected using an estrogen responsive CAT reporter plasmid. Estrogenic activities of photosynthetic algae spirulina and sea lettuce were evaluated using this system. Estrogenic activities of a $500\mug/ml\; and\; 50 \mug/ml$ ethanol extracts of spirulina were as much as that of $10^{-8}$M standard solution (17$\beta$-estradiol) and activity of $5\mug/ml$ ethanol extract of spirulina was as much as that of $10^{-10}$ M standard solution. However, no significant estrogenic activity was observed using sea lettuce extract. Estrogenic activities of marine animals, such as star fish and shrimp, were also evaluated using this system, however, no significant estrogenic activity was observed in these extracts. In this result, it is confirmed that spirulina extract possesses estrogenic compound.

Establishment of In vitro Detection System for iNOS Expression and the Verification of Suppressive Effect by Pine Needle Extract (iNOS 발현 검출을 위한 in vitro 시스템의 확립 및 적송잎 추출물에 의한 저해효과 검증)

  • Kim, Nam-Young;Jang, Hye-Ji;Lee, Dong-Geun;Jang, Min-Kyung;Lee, Seung-Woo;Jeon, Myong-Je;Kim, Mi-Hyang;Kim, Sung-Gu;Lee, Sang-Hyeon
    • KSBB Journal
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    • v.26 no.2
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    • pp.172-176
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    • 2011
  • This study was aimed to verify suppressive effect of pine-needle extract on lipopolysaccharide (LPS)-induced inducible nitric oxide synthase (iNOS) expression. In order to evaluate suppressive effect on iNOS expression, RAW 264.7 cells were stably transfected using an iNOS promoterluciferase reporter plasmid yielding RAW 264.7/pGL2-NeomiNOS_ pro11 cells. Established in vitro detection system revealed to diminish LPS-induced iNOS expression by 0.1~500 ${\mu}g/mL$ of saponin at the concentration-dependant manner. Pine needle extract also diminished LPS-induced iNOS expression to 92 and 88% at 500 and 50 ${\mu}g/mL$, respectively. These results suggest that the in vitro detection system developed here could be useful for the verification of suppressive materials on iNOS expression and pine needle extract could be used for the development of functional foods.

The Isolation of Acetylcholinesterase Inhibitory Constituents from Lycoris radiata using On-line HPLC-biochemical Detection System

  • Yang, Hee-Jung;Yoon, Kee-Dong;Chin, Young-Won;Kim, Young-Choong;Kim, Jin-Woong
    • Natural Product Sciences
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    • v.16 no.4
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    • pp.228-232
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    • 2010
  • Bioactivity-guided fractionation using on-line HPLC biochemical detection system on $CHCl_3$-soluble fraction of Lycoris radiata led to the isolation of deoxylycorenine (1), O-demethylhomolycorine (2), galanthamine (3), lycoramine (4), mixture of $6{\alpha}$-and $6{\beta}$-haemanthidine (5), and lycorine (6), identified by spectroscopic data and physicochemical property. Among the isolated compounds, 1, 3 and 6 showed acetylcholinesterase inhibitiory activities with $IC_{50}$ values of 18.0, 12.0 and $16.6\;{\mu}M$, respectively, in in vitro colorimetric microplate assay.

Evaluation of the Genetic Toxicity of Synthetic Chemicals (XVI) - in vitro Mouse Lymphoma Assay with 3 chemicals -

  • Kim, Youn-Jung;Ryu, Jae-Chun
    • Molecular & Cellular Toxicology
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    • v.2 no.4
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    • pp.244-250
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    • 2006
  • The detection of many synthetic chemicals used in industry that may pose a genetic hazard in our environment is of great concern at present. Since these substances are not limited to the original products, and enter the environment, they have become widespread environmental pollutants, thus leading to a variety of chemicals that possibly threaten the public health. In this respect, to regulate and to evaluate the chemical hazard will be important to environment and human health. The genotoxicity of 3 synthetic chemicals was evaluated in L5178Y $tk^{+/-}$ mouse lymphoma cells in vitro. 9H-carbazole (CAS No. 86-74-8) did not induce significant mutation frequencies both in the presence and absence of metabolic activation system. 1, 3-Dichloro-2-propanol (CAS No. 96-23-1) revealed a significant increase of mutation frequencies in the range of $625-373\;{\mu}g/mL$ in the absence of metabolic activation system and $157-79\;{\mu}g/mL$ in the presence of metabolic activation system. And also, fenpropathrin (CAS No. 64257-84-7) appeared the positive results only in the absence of metabolic activation system. Through the results of MLA tk assay with 3 synthetic chemicals in L5178Y cells in vitro, we may provide the important clues on the genotoxic potentials of these 3 chemicals.

Development of Virtual Endoscopy and Evaluation of Performance as a 3D Virtual Colonoscopy (가상내시경의 개발 및 가상 대장내시경으로 적용 시 성능평가)

  • 김정훈;이상훈;고성호;김상준
    • Journal of Biomedical Engineering Research
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    • v.24 no.2
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    • pp.69-75
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    • 2003
  • Virtual colonoscopy is one of the Powerful tool for non-invasive colon examination and many in-vitro and in-vivo studies have shown its accuracy in Polyp or adenoma detection. But most of virtual colonoscopy requires high quality workstation and software and its cost is high to setup whole system. We developed PC-based 3D model creation and navigation program which has diverse functions. It can be easily installed to PC and connected to network system. The performance. when used as a virtual colonoscopy. is evaluated by calculating sensitivity of detection for the simulated polyp which is artificially made inside the Pig's colon and checked its clinical feasibility, Its total sensitivity is 76%. Grouping according to Polyps diameter, the sensitivity for detection of polyps 10 ㎜ or larger was 100%(40 of 40); 5.0-9.9 ㎜, 90.0(90 of 100): and smaller then 5 ㎜. 36.7%(22 of 60).

Evaluation of the Genetic Toxicity of Synthetic Chemicals (IV) - in vitro Chromosomal Aberration Assay with 18 Chemicals in Chinese Hamster Lung Cells -

  • Ryu, Jae-Chun;Kim, Kyung-Ran;Kim, Youn-Jung
    • Environmental Mutagens and Carcinogens
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    • v.22 no.3
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    • pp.149-156
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    • 2002
  • The detection of many synthetic chemicals used in industry that may pose a genetic hazard in our environment is of great concern at present. Since these substances are not limited to the original products, and enter the environment, they have become widespread environmental pollutants, thus leading to a variety of chemicals that possibly threaten the public health. In this respect, to regulate and to evaluate the chemical hazard will be important to environment and human health. The clastogenicity of 18 synthetic chemicals was evaluated in Chinese hamster lung fibroblast cells in vitro. 4-Chloro-3,5-dimethyl phenol (CAS No. 88-04-0) induced chromosomal aberrations with significance at the concentration of 15.7 $\mu\textrm{g}$/$m\ell$ both in the presence and absence of metabolic activation system. Phenoxybenzene (CAS No. 101-84-8) which is one of the most cytotoxic chemical among 18 chemicals tested revealed no clastogenicity in the range of 0.11-0.43 $\mu\textrm{g}$/$m\ell$ both in the presence and absence of metabolic activation system. From the results of chromosomal aberration assay with 18 synthetic chemicals in Chinese hamster lung cells in vitro, 4-chloro-3,5-dimethyl phenol (CAS No. 88-04-0) revealed weak positive clastogenic results in this study.

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Evaluation of the Genetic Toxicity of Synthetic Chemicals (XIV)-in vitro Chromosomal Aberration Assay with 11 Chemicals in Chinese Hamster Lung Cells

  • Kim, Youn-Jung;Ryu, Jae-Chun
    • Molecular & Cellular Toxicology
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    • v.2 no.2
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    • pp.89-96
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    • 2006
  • The detection of many synthetic chemicals used in industry that may pose a genetic hazard in our environment is of great concern at present. Since these substances are not limited to the original products, and enter the environment, they have become widespread environmental pollutants, thus leading to a variety of chemicals that possibly threaten the public health. In this respect, to regulate and to evaluate the chemical hazard will be important to environment and human health. The clastogenicity of 11 synthetic chemicals was evaluated in Chinese hamster lung fibroblast cells in vitro. 1-Chloro-3-bromopropane CAS No. 109-70-6) induced chromosomal aberrations with significance at the concentration of $185.0\;{\mu}g/mL\;and\;1,600\;{\mu}g/mL$ both in the presence and absence of metabolic activation system, respectively. Triphenyl phosphite (CAS No. 101-02-0), which is one of the most cytotoxic chemical among 11 chemicals tested revealed no clastogenicity in the range of $95.0-4.9\;{\mu}g/mL$ both in the presence and absence of metabolic activation system. From the results of chromosomal aberration assay with 11 synthetic chemicals in Chinese hamster lung cells in vitro, 1-chloro-3-bromopropane revealed a positive clastogenic result in this study.

Detection of Testosterone-like Activities of Hydrothermal and Ethanol Extracts of Oriental Medicines Using an in Vitro Detection System (In vitro 검출 시스템을 이용한 한약재 열수 및 에탄올 추출물의 남성호르몬 유사활성 검출)

  • Lee, Dong-Geun;Jo, Jung-Kwon;Lee, Sang-Hyeon
    • Journal of Life Science
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    • v.30 no.6
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    • pp.550-554
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    • 2020
  • The purpose of this study was to screen the testosterone-like activities of eight oriental medicines with an in vitro testosterone compound detection (TCD) system. Distilled water and 80% ethanol, both at 80℃, were used to obtain extracts from Cervus nippon temminck (Nokgaksang), Cynanchum wilfordii (Baeksuo), Lespedeza cuneata (Yagwanmun), Panax ginseng (Hongsam), Smilax china (Toryeong), Taraxacum platycarpum (Mindlre), Tribulus terresteis (Namgase), and Trigonella foenum-graecum (Horopa), and extracts at concentrations of 5, 50, and 500 ㎍/ml were assessed using the TCD system. The testosterone-like activities of the 80% ethanol extracts were seen to increase with concentration and ranged from 0.9 to 5.0 times higher than those of the negative control; Horopa and Yagwanmun exhibited superior testosterone-like activities, followed by Mindlre and Hongsam at the same concentration. Toryeong and Namgase, on the other hand, showed low activity with Nokgaksang and Baeksuo exhibiting very low levels. Compared with the activities of the positive control (5α-DHT), the 80% ethanol extracts of Horopa and Yagwanmun at a concentration of 500 ㎍/ml showed higher testosterone-like activities than 10-8 M 5α-DHT. The water extracts showed lower activity levels than the ethanol samples, and the change of activity with concentration was also lower. It was confirmed that the 80% ethanol extracts of Horopa, Yagwanmun, and Mindlre etc. enhance the transcription efficiency of testosterone-related genes and may therefore be useful in developing health food or medicinal treatments for the improvement of male menopausal symptoms.