• Title/Summary/Keyword: in vitro bioassay

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Application of Single Cell Gel Electrophoresis for Detection of DNA Single Strand Breaks in DNA of Fish Blood Cell (어류혈구세포에 있어서 Single Cell Gel Electrophoresis를 응용한 DNA Single Strand Breack의 측정)

  • KIM Gi Beum;LEE Richard F.;MARUYA Keith A.
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.36 no.4
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    • pp.346-351
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    • 2003
  • Single-cell gel electrophoresis (comet assay) was used to detect DNA single strand break in blood cells from several marine fish species. Three fish species were collected from Georgia coastal area. Mummichog, Fundulus heteroclitus showed higher DNA damage than sea bass, Lateolabrax japonicus and trout, Oncorhynchus masou masou under the same experimental conditions. Mummichogs had more alkaline-labile sites on their DNA than other fish species. The comet assay with mummichog blood cells at pH 12.5 showed a dose-response curve with the increasing concentrations of hydrogen peroxide. While the isolated leucocytes showed no increase of DNA damage after in vitro exposure to 2-methyl-1,4-naphthoquinone (MNQ), erythrocytes showed dose-dependent DNA damage. These results indicate that the comet assay can be applied successfully as a bioassay using erythrocyte for environmental monitoring.

Preclinlcal Evaluation of Polysaccharides Extracted from Korean Red-ginseng as an Antineoplastic Immunostimulator (홍삼다당체의 항암면역증강작용 연구)

  • 김기환;정인성
    • Journal of Ginseng Research
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    • v.21 no.2
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    • pp.78-84
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    • 1997
  • As a part of our ongoing effort to develop new antineoplastic immunostimulator from natural sources, bioassay-directed fractionationn of polysaccharides from Korean red ginseng was carried out by observing the proliferation of marine spleen cells and the generation of lymphoklne activated killer (LAK) cells. The acidic polysaccharide fractions proliferated spleen cells and generated LAK cells in proportion to their acidity in vitro. The LAK cell which was induced by ginseng showed tumoricidal activity against both NK celt sensitive and insensitive tumor target cells without major histocompatibility (MHC) restriction. Adherent macrophages and CD4+helper T cells were involved in the generation of the LAK cells. The acidic polysaccharide from Korean red ginseng synerglzed with recombinant IL-2 (rIff-2) at lower than 3 U/ml. The optimal doses of the acidic polysaccharide from Korean red ginseng for the proliferation of spleen cells and for the generation of LAK cells were 1 mg/ml and 100 $\mu\textrm{g}$/ml, respectively; this means that the mechanisms for the both activities may be different from each other.

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Antagonism of Pseudomonas spp. against to Rhizoctonia solani and Pythium spp. (Pseudomonas spp.의 Rhizoctonia solani 및 Pythium spp. 병원균에 대한 길항작용)

  • 주영규;한정훈
    • Asian Journal of Turfgrass Science
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    • v.8 no.1
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    • pp.47-52
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    • 1994
  • Attempts were made to investigate the antagonistic activity of soil borne microorganisms Pseudomonas spp. and Trichoderma spp. against to the pathogens of turf diseases Rhizoctionia solani spp. and Pythiom spp. in vitro by a dual culture bioassay. Inhibition zone between the edge of the my-celium and the margin of each antagonistic bacteria, Pocudontonas, on potato dextrose agar was measured 3 days after incubation at 28˚C. Psudomonas spp. showed relatively high inhibition of mycelium growth of R. solani AG-i and Pythium spp. which cause brown patch and pythium blight, respectively. Antagonistic fungi Trichodenma spp. also showed effective inhibition against mycelium growth of both pathogens, more proper methods of measuring the inhibition effects were required because of fast growth of Trichodenna hypae. Brown patch and pythium blight both, re-quire most higher rate of fungicide use to control in golf curses in Korea. Application of antagon-istic microorganisms are useful as biological resources an approach to sole environmental contamination.

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Identification of Anti-Cancer Targets of Eco-Friendly Waste Punica granatum Peel by Dual Reverse Virtual Screening and Binding Analysis

  • Usha, Talambedu;Goyal, Arvind Kumar;Lubna, Syed;Prashanth, H.P.;Mohan, T. Madhan;Pande, Veena;Middha, Sushil Kumar
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.23
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    • pp.10345-10350
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    • 2015
  • Background: Punica granatum (family: Lythraceae) is mainly found in Iran, which is considered to be its primary centre of origin. Studies on pomegranate peel have revealed antioxidant, anti-inflammatory, anti-angiogenesis activities, with prevention of premature aging and reducing inflammation. In addition to this it is also useful in treating various diseases like diabetes, maintaining blood pressure and treatment of neoplasms such as prostate and breast cancer. Objectives: In this study we identified anti-cancer targets of active compounds like corilagin (tannins), quercetin (flavonoids) and pseudopelletierine (alkaloids) present in pomegranate peel by employing dual reverse screening and binding analysis. Materials and Methods: The potent targets of the pomegranate peel were annotated by the PharmMapper and ReverseScreen 3D, then compared with targets identified from different Bioassay databases (NPACT and HIT's). Docking was then further employed using AutoDock pyrx and validated through discovery studio for studying molecular interactions. Results: A number of potent anti-cancerous targets were attained from the PharmMapper server according to their fit score and from ReverseScreen 3D server according to decreasing 3D scores. Conclusion: The identified targets now need to be further validated through in vitro and in vivo studies.

Differential antiangiogenic and anticancer activities of the active metabolites of ginsenoside Rg3

  • Maryam Nakhjavani;Eric Smith;Kenny Yeo;Yoko Tomita;Timothy J. Price;Andrea Yool;Amanda R. Townsend;Jennifer E. Hardingham
    • Journal of Ginseng Research
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    • v.48 no.2
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    • pp.171-180
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    • 2024
  • Background: Epimers of ginsenoside Rg3 (Rg3) have a low bioavailability and are prone to deglycosylation, which produces epimers of ginsenoside Rh2 (S-Rh2 and R-Rh2) and protopanaxadiol (S-PPD and R-PPD). The aim of this study was to compare the efficacy and potency of these molecules as anti-cancer agents. Methods: Crystal violet staining was used to study the anti-proliferatory action of the molecules on a human epithelial breast cancer cell line, MDA-MB-231, and human umbilical vein endothelial cells (HUVEC) and compare their potency. Cell death and cell cycle were studied using flow cytometry and mode of cell death was studied using live cell imaging. Anti-angiogenic effects of the drug were studied using loop formation assay. Molecular docking showed the interaction of these molecules with vascular endothelial growth factor receptor-2 (VEGFR2) and aquaporin (AQP) water channels. VEGF bioassay was used to study the interaction of Rh2 with VEGFR2, in vitro. Results: HUVEC was the more sensitive cell line to the anti-proliferative effects of S-Rh2, S-PPD and R-PPD. The molecules induced necroptosis/necrosis in MDA-MB-231 and apoptosis in HUVEC. S-Rh2 was the most potent inhibitor of loop formation. In silico molecular docking predicted a good binding score between Rh2 or PPD and the ATP-binding pocket of VEGFR2. VEGF bioassay showed that Rh2 was an allosteric modulator of VEGFR2. In addition, SRh2 and PPD had good binding scores with AQP1 and AQP5, both of which play roles in cell migration and proliferation. Conclusion: The combination of these molecules might be responsible for the anti-cancer effects observed by Rg3.

Isolation of Ethanol Metabolizing Enzyme Inhibitors from Aloe spp.

  • Shin, Kuk-Hyun;Woo, Won-Sick;Chung, Ha-Sook;Shim, Chang-Sub
    • Natural Product Sciences
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    • v.1 no.1
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    • pp.55-60
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    • 1995
  • In the course of evaluation of hepatoprotective components against alcohol-induced toxicity from Aloe spp., the methanol extract was found to cause a significant inhibition of rat liver cytosolic alcohol dehydrogenase activity. Systematic fractionation of active tractions monitored by bioassay led to isolation of four compounds; aloe-emodin, aloenin, ethylidene-aloenin and ${\beta}-sitosterol$, which were estimated as active principles for inhibition of c-ADH and c-ALDH activities in vitro.

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Isolation and Evaluation of an Antitumor Constituent from Pyrolae Herba (녹수초의 항암활성물질의 분리 및 항암력 평가)

  • Bae, Ki-Hwan;Kim, Hwan-Mook;Lee, Sang-Myung
    • YAKHAK HOEJI
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    • v.40 no.2
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    • pp.225-229
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    • 1996
  • The cytotoixic effect of Pylorae Herba (Pyrola japonica Klenze) against L1210 and K-562 cells was studied in vitro. The methanolic extract of Pylorae Radix was add ed to the culture of L1210 cells and K-562 cells for the cytotoxic activity and the ED50 values of hexane, ethylacetate, buthanol and water fractions from methanolic extract were determined using MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazoliumbromide) assay. The active constituent isolated by bioassay guided fractionation followed by purification gave rise to a yellow needle crystal and was clarified to be chimaphilline by the comparison with the published data. The average life spans with it were not prolonged significantly on tumor growth in hybrid female mouse (BDF1-KIST) inoculated subcutaneously with P388 cells.

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Phytochemical Constituents of Geranium eriostemon

  • Chang, Sang-Wook;Kim, Ki-Hyun;Lee, Il-Kyun;Choi, Sang-Un;Lee, Kang-Ro
    • Natural Product Sciences
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    • v.15 no.3
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    • pp.151-155
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    • 2009
  • Phytochemical investigation of the MeOH extract of the aerial parts of Geranium eriostemon resulted in the isolation of one triterpene, three furofuran lignans, one syringic acid and four flavonoids. Their chemical structures were characterized by spectroscopic methods to be oleanolic acid (1), (-)-kobusin (2), (-)-eudesmin (3), (+)-magnolin (4), syringic acid (5), quercetin (6), juglanin (7), juglalin (8), and hyperin (9). All compounds (1 - 9) were isolated for the first time from this plant source and the compounds 2 - 4 were reported first from the genus Geranium. Compounds 4 - 6 exhibited moderate cytotoxicity against four human cancer cell lines in vitro using a SRB bioassay.

Inhibitory Effect of GE974 isolated from Gyrophora esculenta on ${\alpha}-Glucosidase$ (석이에서 분리한 GE974의 ${\alpha}-Glucosidase$ 저해효과)

  • Choi, Hyuck-Jai;Kim, Dong-Hyun;Kim, Nam-Jae
    • Korean Journal of Pharmacognosy
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    • v.31 no.2
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    • pp.196-202
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    • 2000
  • This study was conducted to search for the ${\alpha}-glucosidase$ inhibitor from the natural products. In the previous study, the water extract of Gyrophora esculenta exhibited a potent inhibitory effect on ${\alpha}-glucosidase$ activities. Then, by bioassay-guided fractionation followed by chromatographic separation of the water extract of Gyrophora esculenta, ${\alpha}-glucosidase$ inhibitor was isolated as GE974. GE974 showed significant inhibitory activities on some kinds of ${\alpha}-glucosidase$ in vitro. Its inhibitory mechanism seemed to be competitive for disaccharides. Also, it markedly inhibited ${\alpha}-glucosidases$ of intestine separated from both nondiabetics and diabetics.

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Monoamine Oxidase B Inhibitors from the Fruits of Opuntia ficus-indica var. saboten

  • Han, Yong-Nam;Choo, Yeun-Su;Lee, Young-Chul;Moon, Young-In;Kim, Sung-Dae;Choi, Jong-Won
    • Archives of Pharmacal Research
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    • v.24 no.1
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    • pp.51-54
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    • 2001
  • Three varieties of methyl citrate and 1 -methyl malate were isolated from the fruits of Opuntia ficus-indica var. saboten Makino through in vitro bioassay-guided isolation for the inhibition on monoamine oxidase(MAO). The $IC_50$ values for MAO-B of 1-monomethyl citrate, 1,3-dimethy citrate, trimethyl citrate and 1-methyl malate were 0.19, 0.23, 0.61 and 0.25 mM, respectively. However, on MAO-A, their inhibitions showed only marginal activity.

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