• Title/Summary/Keyword: sulforhodamine B (SRB) method

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Use of cccDNA Breakage Assay and Sulforhodamine B Assay for the Prescreening of Antitumor Agents from Microbial Sources (미생물 배양액으로부터 항암제의 예비선별을 위한 cccDNA Breakage 활성검정과 Assay Sulforhodamine B 활성검정의 이용)

  • Lee, Sang-Han;Lee, Dong-Sun;Kim, Jong-Guk;Hong, Soon-Duck
    • Journal of Life Science
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    • v.8 no.1
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    • pp.67-71
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    • 1998
  • In order to develop new antitumor agents from fermentation broths, we used cccDNA breakage assay abd sulforhodamine B assay for prescreening. As a result, it was shown that sample reach 3.3% when using cccDNA breakage assay. In sulforhodamine B assay, we obtained 4 acive fraction against A549 (a cell line of human lung carcinoma) and SK-OV-3 (a cell line of human adenocarcinoma). These results suggest that these assay would be a promising method for antitumor prescreening from microbial sources.

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Sulforhodamine B Assay to Determine Cytotoxicity of Vibrio vulnificus Against Human Intestinal Cells

  • Lee, Byung-Cheol;Choi, Sang-Ho;Kim, Tae-Sung
    • Journal of Microbiology and Biotechnology
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    • v.14 no.2
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    • pp.350-355
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    • 2004
  • Sulforhodamine B (SRB) assay is a rapid, sensitive, and inexpensive method for measuring cell proliferation and chemosensitivity. However, the lactate dehydrogenase (LDH) release assay is generally used to measure cytototoxicity of infectious microorganisms against host cells. In this study, we investigated the possibility of applying the SRB assay to determine cytotoxicity for infectious microorganisms, and compared the results with those obtained by the LDH release assay. We used Vibrio vulnificus as a model of infectious microorganisms. The SRB assay showed that V vulnificus strongly induced cytotoxic activity against human intestinal cells, Caco-2 and INT-407 cells. The degree of cytotoxicity closely correlated with infection time and number ratios of V. vulnificus to intestinal cells (MOI, multiplicity of infection). Furthermore, cytotoxicity values obtained by SRB assay correlated well with results obtained by the LDH release assay, and both assays gave a linear response with respect to MOI Heat-inactivation of V. vulnificus for 35 min at $60^{\circ}C$ did not induce cytotoxic activity, indicating that viability of V. vulnificus is crucial for cytotoxic activity against intestinal cells. Although both assays are suitable as cytotoxicity endpoints, the SRB assay is recommended for measuring cytotoxicity of infectious microorganisms against host cells because of its significantly lower cost and more stable endpoint than the LDH release assay.

Anti-influenza Compounds Isolated from Descurainia sophia Seeds

  • Woo Seung Yang;Choong Je Ma
    • Natural Product Sciences
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    • v.29 no.2
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    • pp.113-119
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    • 2023
  • Descurainia sophia seeds methanol extract showed significant anti-influenza activity and we tried to isolate anti-influenza compounds from the D. sophia extract. D. sophia seeds were extracted with 80% methanol and fractionated with n-hexane, ethyl acetate, CHCl3 and n-butanol. The anti-influenza activity of each fraction was assessed using sulforhodamine B (SRB) method in A549 cells, human-derived lung cancer cells. The ethyl acetate and CHCl3 fractions showed the most potent anti-influenza activity. Seven compounds were isolated from CHCl3 fraction and identified 1-decanol (1), 2-(3,4-dihydroxy-2-methylenebutoxy)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol (2), daucosterol (3), isorhamnetin (4), quercetin (5), sinapic acid (6), and helveticoside (7) by spectroscopic data such as UV, IR, 1H-NMR, 13C-NMR and mass spectroscopy. Anti-influenza activities of isolated compounds were evaluated using SRB method in A549 cells. Compounds 3, 4 and 7 had significant anti-influenza activity in a dose-dependent manner.

The Inhibitory Effects of Taraxaci Herba against Cadmium induced Cytotoxicity (포공령의 카드뮴에 대한 세포독성 억제효과)

  • Han, Du-Seok;Lee, Ki-Nam;Lee, Jong-Sub;Baek, Seung-Hwa
    • YAKHAK HOEJI
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    • v.42 no.3
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    • pp.307-311
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    • 1998
  • This study was carried out to evaluate antitoxic effects Taraxaci Herba extract against Cadium by calorimetric methods. The antitoxic activity of Taraxaci Herba ex tract in NIH 3T3 fibroblasts was evaluated by MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-phenyl-2H-tetrazoliumbromide), NR (Neutral red) and SRB (Sulforhodamine B protein) assay. The light microscopic study was carried out to observe morphological changes of the treated cells. These results were obtained as follows; The concentration of $10^{-2}mg/ml$ of Taraxaci Herba extract was shown significant antitoxic activity. The number of NIH 3T3 fibroblasts were antitoxic and tend to regenerate. These results suggest that Taraxaci Herba extract retains a potential antitoxic activity.

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Cytotoxicity of Ligularia fischeri Extracts (곰취 추출물의 세포독성 효과)

  • 함승시;이상영;오덕환;정성원;김상헌;정차권;강일준
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.27 no.5
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    • pp.987-992
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    • 1998
  • This study was investigated to observe the cytotoxicity effect of Ligularia fischeri extracts against cancer cell lines including human lung carcinoma(A549), human cervix epitheloid carcinoma(HeLa) and human hepatocellular carcinoma(HepG2) using SRB(sulforhodamine B) method. The ethanol and methanol extracts of 1$\mu\textrm{g}$/${mu}ell$ showed approximately 79.2% and 86.4% cytotoxicity effects on HepG2 cell line and the ethyl acetate fracton fractionated from ethanol extracts showed the strongest cytotoxicity effect with 94% inhibition. The inhibitory effect of ethanol extract on HeLa cell line was somewhat low with 50~56% inhibition, but ethyl acetate fraction showed higher cytotoxicity effect with 91% and 91.9% inhibition on the HeLa and A549 cell line. On the contrary, the ethanol and methanol extracts showed the lower inhibition effects on the normal liver cell, WRL68, compared to human cancer cell lines.

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Development of Liposome Immunoassay for Salmonella spp. using Immunomagnetic Separation and Immunoliposome

  • Shin, Jung-Hee;Kim, Myung-Hee
    • Journal of Microbiology and Biotechnology
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    • v.18 no.10
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    • pp.1689-1694
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    • 2008
  • The ability to detect Salmonella spp. is essential in the prevention of foodborne illness. This study examined a Salmonella spp. detection method involving the application of immunomagnetic separation and immunoliposomes (IMS/IL) encapsulating sulforhodamine B (SRB), a fluorescent dye. A quantitative assay was conducted by measuring the fluorescence intensity of SRB that was produced from an immunomagnetic bead-Salmonella spp.-immunoliposome complex. The results indicated detection limits of $2.7{\times}10^{5}$ and $5.2{\times}10^{3}$ CFU/ml for Salmonella enterica subsp. enterica serovar Enteritidis (S. Enteritidis) and Salmonella enterka subsp. enterka serovar Typhimurium (S. Typhimurium), respectivley. The signal/noise ratio was improved by using 4% skim milk as a wash solution rather than 2% BSA. In addition, higher fluorescence intensity was obtained by increasing the liposome size. Compared with the conventional plating method, which takes 3-4 days for the isolation and identification of Salmonella spp., the total assay time of to h only including 6 h of culture enrichment was necessary for the Salmonella detection by IMS/IL. These results indicate that the IMS/ IL has great potential as an alternative rapid method for Salmonella detection.

The Growth Inhibitory Effects of Epigallocatechin Gallate Against Human Skin Melanoma Cells and Human Oral Epitheloid Carcinoma Cells (Epigallocatechin gallate의 인체 피부흑색종세포와 인체 구강유상피암종세포에 대한 성장억제효과)

  • 한두석;박승택;백승화
    • Environmental Mutagens and Carcinogens
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    • v.18 no.2
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    • pp.98-103
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    • 1998
  • Epigallocatechin gallate (EGCG) was reported to exert weak cytotoxicity against normal healthy cells such as C3H10T1/2 cells, but profound inhibitory effects on the initiation or promotion stage of chemical carcinogenesis in mammary gland, blood and mouse skin. This study was carried out to develop antitumor agents with weak side effects and strong antitumor activity. Human skin melanoma cells (HBT 69) and human oral epitheloid carcinoma cells (OCL 17) were cultured in RPMI-1640 media containing 10% fetal bovine serum, antibiotic, and fungizone. After incubation for 24 hrs, the cells were treated with various amounts of (EGCG) for 48 hrs. The growth inhibitory effects of EGCG in human oral epitheloid carcinoma cells were evaluated by the 3- (4,5-djmethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT), neutral red (NR), and sulforhodamine B protein (SRB) assays of colorimetric methods. The light microscopic study was also carried out to observe morphological changes of the treated cells. These results obtained were as follows; 1. Significantly inhibitory effects of EGCG against cultured human oral epithelioid carcinoma cells. 2. Significantly inhibitory effects against cultured human skin melanoma cells treated with 50 $\mu$M EGCG, but decreased inhibitory effects in 100 $\mu$M EGCG. 3. Degenerative changes against cultured human oral epitheloid carcinoma cells. 4. Degenerative changes against human skin melanoma cells treated with 50 UM EGCG, but recovered degenerative changes in 100 $\mu$M EGCG.

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Screening for Inhibitor of c-myc Expression and Identification of Isolate No.2303

  • Chung, Ji-Hyung;Yeo, Ick-Hyun;Oh, Doo-Whan;Moon, Soon-Ok
    • Journal of Microbiology and Biotechnology
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    • v.5 no.5
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    • pp.264-268
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    • 1995
  • Sulforhodamine B(SRB) assay was performed on the human lung carcinoma, A549 cell line to screen soil microorganisms for production of anti-cancer agent. Among 4, 265 microorganisms, 45 isolates were selected for their cytotoxicity and tested for their effects on the expression of c-myc by RNA slot blot and Northern blot analysis resulting in selection of No.2303 isolate. This No.2303 was identified as Streptomyces sp. by ISP classification and the chemotaxonomic analysis method. NO.2303 inhibited the expression of cmyc in Col0320 DM and A549 cell lines. The culture extract of No. 2303 also inhibited the progression of the cell cycle of Go in NIH 313 cells, implying that the extract also inhibited the expression of c-myc in NIH 313 cell.

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Cytotoxic Effect of the Pine needle extracts (솔잎 추출물의 in vitro계 암세포 성장억제효과)

  • Kim, Eun-Jeong;Jung, Sung-Won;Choi, Keun-Pyo;Ham, Seung-Shi;Gang, Ha-Yeong
    • Korean Journal of Food Science and Technology
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    • v.30 no.1
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    • pp.213-217
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    • 1998
  • This study was performed to observe cytotoxic effect of the pine needle extracts against cancer cell lines including human gastric carcinoma (KATOIII), human lung carcinoma (A549), human hepatocellular carcinoma (Hep3B) and human breast adenocarcinoma (MCF-7) using MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide) and SRB (sulforhodamine B) method. The extracts were prepared by step-wise fractionation of ethanol extract of pine needles using diethylether, chloroform, ethylacetate, butanol and water. The growth of the cancer cells in medium containing pine needle extracts were significantly inhibited degree in proportion to the increase of the extract concentration. A significant shrinkage of Hep3B cells was observed when the cells were exposed into 0.5, 1 mg/mL of pinus rigida extract.

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A Study of Cytotoxicity from Some Korean Edible Plants (수종 한국산 식용식물의 세포독성 연구)

  • 정하숙
    • Korean journal of food and cookery science
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    • v.15 no.2
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    • pp.108-113
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    • 1999
  • Natural products derived from not only medicinal but edible plants have been used as sources of folk remedies and other useful materials, like as appetizers, health supplements and food additives. A short-term in vitro biomarker assay was accompilshed to assess cytotoxic activity on the human lung and ovary adeno cancer cell lines based on sulforhodamine B (SRB) method. As a result, the EtOAc soluble fractions from Trichosanthes kirilowii Max. and Dioscorea japonica Thunb. showed potent cytotoxicity as a below 30% of growth ratio of cancer cell at a concentration of 40 $\mu\textrm{g}$/ml on lung and ovary adeno cancer cell lines, and lung cancer cell line, respectively. Cytotoxic activity present in plant extracts appear to be promising candidates as functional foods among Korean wild edible plants, and further studies are warranted.

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