• Title/Summary/Keyword: in vitro assay

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Comparative Studies of Adriamycin and 28-Deacetyl Sendanin on In Vitro Growth Inhibition of Human Cancer Cell Lines

  • Kim, Hwan-Mook;Oh, Goo-Taeg;Han, Sang-Bae;Hong, Dong-Ho;Hwang, Bang-Yeon;Kim, Young-Ho;Lee, Jung-Joon
    • Archives of Pharmacal Research
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    • v.17 no.2
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    • pp.100-103
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    • 1994
  • The limonoid compound (28-deacetyl sendanin0 isolated from the fruit of Melia toosendan SIEB. et ZUCC. was evaluated on anticancer activity. According to a standard in vitro cytotoxicity assy, eight human cancer cell lines and SRB assay were introduced for present evaluation. As a positive standard, adriamycin was tested in parallel. The cell lines were originated from six different organs. In view of dose-response profiles to 28-deacetyl sendanin, the most sensitive cells were SF-539 and PC-3 which were derived from CNS and prostate, respecitively. In contrast, all the cell lines responded similarly to adriamycin to give rise to nearly indentical six cell lines were more sensitive to 28-deacetyl sendanin and two were more resistant. As a result, 28-deacetyl sendanin had more senstive and selective inhibitory effects on in vitro growth of human cancer cell lines in a comparison with adriamycin.

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In Vitro and Cellular Antioxidant Activity of a Water Extract of Saururus chinensis

  • Kim, Gyo-Nam;Lee, Jung-Sook;Jang, Hae-Dong
    • Food Science and Biotechnology
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    • v.17 no.6
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    • pp.1332-1336
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    • 2008
  • The water extract of Saururus chinensis was investigated for oxygen radical absorbance capacity (ORAC), reducing capacity, metal chelating activity, and intracellular antioxidant activity using HepG2 cell. When 2,2'-azobis(2-amidinopropane) dihydrochloride (AAPH) was used for the generation of peroxyl radicals in vitro, S. chinensis extract (SC-E) showed the strong and concentration-dependent scavenging activity through donating protons which could be explained by its reducing property. When hydroxyl radicals were generated in vitro through the addition of $Cu^{2+}$ and $H_2O_2$, SC-E demonstrated the antioxidant activity depending on its concentration. In HepG2 cell model, most of intracellular oxidative stress generated by AAPH was efficiently removed by SC-E. However, when $Cu^{2+}$ without $H_2O_2$ was used as an oxidant in the intracellular assay, SC-E partially reduced the oxidative stress caused by $Cu^{2+}$ in cellular antioxidant activity assay system. These results indicate that SC-E could be utilized for the development of functional foods as antioxidant resource in the near future.

Detection of Anticancer Activity from the Root of Angelica gigas In Vitro

  • Ahn, Kyung-Seop;Sim, Woong-Seop;Kim, Ik-Hwan
    • Journal of Microbiology and Biotechnology
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    • v.5 no.2
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    • pp.105-109
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    • 1995
  • Anticancer activity of a fraction of the ethanol extract from the root of Korean angelica (Angelica gigas Nakai) was recognized in human cancer cell lines HeLa $S_3$, K-562, and Hep $G_2$. The extract blocked the phorbol ester-inducing megakaryocytic differentiation of K-562 cells, which indicated the modification of protein kinase C (PKC) activity. In vitro assay showed the activation of PKC by the extract. An effective fraction of the Angelica gigas extract, of which $R_f$ value was 0.64 in a thin layer chromatography, was a different component from those of European angelicas. The $ED_50$ value of the fraction was 8, 9, and $16\;\mu\textrm{m}/ml$ against HeLa $S_3\;Hep\;G_2$, and K-562 cells, respectively, while the fraction showed higher $ED_50$ values against normal cell lines.

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Antioxidant and Anticancer Activities of Extract from Artemisia capillaries (인진쑥 추출물의 항산화 및 항암 활성)

  • Jung, Mee-Jung;Yin, Yu;Heo, Seong-Il;Wang, Myeong-Hyeon
    • Korean Journal of Pharmacognosy
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    • v.39 no.3
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    • pp.194-198
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    • 2008
  • Artemisia capillaries is a major important food and medicinal resource in Korea. In order to confirm the biological activities of Artemisia capillaries, we investigated antioxidant and anticancer activities from in vitro assays. The Artemisia capillaries methanol (MeOH) extracts was used for the evaluation of DPPH scavenging, total phenolic content, total flavonoid content, hydroxyl radical (${\bullet}OH$) scavenging, reducing power assay as antioxidant activity, as well as anticancer activities as MTT assay. As a result, the Artemisia capillaries MeOH extracts showed potent antioxidative activity and anticancer activity in vitro. These results suggest that the Artemisia capillaries MeOH extracts have a potential alleviated oxidation process, cell motility activity, and tumorigenesis.

In Vitro Mammalian Chromosomal Aberration Test of Allyl Chloride for Workers' Health (근로자의 건강보호를 위한 알릴 염화물의 포유류 배양세포 염색체이상시험)

  • Rim, Kyung-Taek;Kim, Soo-Jin
    • Journal of Korean Society of Occupational and Environmental Hygiene
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    • v.24 no.2
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    • pp.160-168
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    • 2014
  • Objectives: Chemical hazard evaluations are important for workers' health and working environments. Allyl chloride (CAS No. 107-05-1) is used in many industries, leading to concerns about the possibility of threats to the health of workers. Since only insufficient or controversial information is available about potential related hazards, an in vitro mammalian chromosomal aberration (CA) assay was conducted in order to gain additional information concerning any such hazards. Moreover, toxicological information from this study could be applied for workers' rights to know, and to prepare or update the Materials Safety Data Sheet (MSDS) for a number of industries. Methods and Results: The assay was performed using the Chinese hamster lung fibroblast cell (ATCC, CRL-1935), by the direct method (-S9) and by the metabolic activated method (+S9 mix). Using the direct method, the seven dosages in the 48-hour treatment group did not show that the frequency of CA is proportionate to the dosage. The frequency of CA is not proportionate to the dosage addition for a six-hour treatment using the metabolic activated method. Conclusions: From these findings, it was decided that this chemical does not induce chromosomal aberrations under the tested conditions.

Lipid Peroxidation and the Thiobarbituric Acid Assay: Standardization of the Assay When Using Saturated and Unsaturated Fatty Acids

  • Rael, Leonard T.;Thomas, Gregory W.;Craun, Michael L.;Curtis, C. Gerald;Bar-Or, Raphael;Bar-Or, David
    • BMB Reports
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    • v.37 no.6
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    • pp.749-752
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    • 2004
  • Saturated fatty acids are less vulnerable to lipid peroxidation than their unsaturated counterparts. In this investigation, individual fatty acids of the $C_{16}$, $C_{18}$ and $C_{20}$ families were subjected to the thiobarbituric (TBA) assay. These fatty acids were chosen based on their degree of saturation and configuration of double bonds. Interestingly, an assay threshold was reached where increasing the fatty acid concentration resulted in no additional decrease in the TBARS concentrations. Therefore, the linear range of TBARS inhibition was determined for fatty acids in the $C_{16}$ and $C_{20}$ families. The rate of TBARS inhibition was greater for the saturated than for unsaturated fatty acids, as measured from the slope of the linear range. These findings demonstrate the need to standardize the TBARS assay using multiple fatty acid concentrations when using this assay for measuring in vitro lipid peroxidation.

Comparing In Vitro and In Vivo Genomic Profiles Specific to Liver Toxicity Induced by Thioacetamide

  • Kang, Jin-Seok;Jeong, Youn-Kyoung;Shin, Ji-He;Suh, Soo-Kyung;Kim, Joo-Hwan;Lee, Eun-Mi;Kim, Seung-Hee;Park, Sue-Nie
    • Biomolecules & Therapeutics
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    • v.15 no.4
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    • pp.252-260
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    • 2007
  • As it is needed to assay possible feasibility of extrapolation between in vivo and in vitro systems and to develop a new in vitro method for toxicity testing, we investigated global gene expression from both animal and cell line treated with thioacetamide (TAA) and compared between in vivo and in vitro genomic profiles. For in vivo study, mice were orally treated with TAA and sacrificed at 6 and 24 h. For in vitro study, TAA was administered to a mouse hepatic cell line, BNL CL.2 and sampling was carried out at 6 and 24 h. Hepatotoxicity was assessed by analyzing hepatic enzymes and histopathological examination (in vivo) or lactate dehydrogenase (LDH) assay and morphological examination (in vitro). Global gene expression was assessed using microarray. In high dose TAA-treated group, there was centrilobular necrosis (in vivo) and cellular toxicity with an elevation of LDH (in vitro) at 24 h. Statistical analysis of global gene expression identified that there were similar numbers of altered genes found between in vivo and in vitro at each time points. Pathway analysis identified several common pathways existed between in vivo and in vitro system such as glutathione metabolism, bile acid biosynthesis, nitrogen metabolism, butanoate metabolism for hepatotoxicty caused by TAA. Our results suggest it may be feasible to develop toxicogenomics biomarkers by comparing in vivo and in vitro genomic profiles specific to TAA for application to prediction of liver toxicity.

Investigation of Chemotactic Activities in Differentiated HL-60 Cells by a Time-lapse Videomicroscopic Assay

  • Jung, Yun-Jae;Woo, So-Youn;Ryu, Kyung-Ha;Jang, Myoung-Ho;Miyasaka, Masayuki;Seoh, Ju-Young
    • IMMUNE NETWORK
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    • v.6 no.2
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    • pp.76-85
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    • 2006
  • Background: Chemotaxis is one of the cardinal functions of leukocytes, which enables them to be recruited efficiently to the right place at the right time. Analyzing chemotactic activities is important not only for the study on leukocyte migration but also for many other applications including development of new drugs interfering with the chemotactic process. However, there are many technical limitations in the conventional in vitro chemotaxis assays. Here we applied a new optical assay to investigate chemotactic activities induced in differentiated HL-60 cells. Methods: HL-60 cells were stimulated with 0.8% dimethylformamide (DMF) for 4 days. The cells were analyzed for morphology, flow cytometry as well as chemotactic activities by a time-lapse videomicroscopic assay using a chemotactic microchamber bearing a fibronectin-coated cover slip and an etched silicon chip. Results: Videomicroscopic observation of the real cellular motions in a stable concentration gradient of chemokines demonstrated that HL-60 cells showed chemotaxis to inflammatory chemokines (CCL3, CCL5 and CXCL8) and also a homeostatic chemokine (CXCL12) after DFM-induced differentiation to granulocytic cells. The cells moved randomly at a speed of $6.99{\pm}1.24{\mu}m/min$ (n=100) in the absence of chemokine. Chemokine stimulation induced directional migration of differentiated HL-60 cells, while they still wandered very much and significantly increased the moving speeds. Conclusion: The locomotive patterns of DMF-stimulated HL-60 cells can be analyzed in detail throughout the course of chemotaxis by the use of a time-lapse videomicroscopic assay. DMF-stimulated HL-60 cells may provide a convenient in vitro model for chemotactic studies of neutrophils.

COMBINED IN VITRO ASSAY FOR 3T3 NRU PT ASSAY AND PHOTOHEMOLYSIS AS PART OF PHOTOTOXICITY TEST

  • Chunja Nam;Kim, Baehwan;Lee, Byoungseok;Seongjoon Moon;Ihseop Chang
    • Proceedings of the Korean Society of Toxicology Conference
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    • 2001.05a
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    • pp.117-117
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    • 2001
  • The aim of this study was to assess a possible alternative method as replacement for in vivo phototoxicity test. The 3T3 mouse fibroblast neutral red uptake phototoxicity assay (3T3 NRU PT assay) is a screening method for studying DNA or cellular damage. Photohemolysis assay is a mechanistic study for investigating oxygen-dependent membrane damage.(omitted)

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Genotoxicological Safety of Hot Water Extracts of the γ-Irradiated Astragali Radix, Atractylodes Rhizoma, and Cimicifugae Rhizoma in Vitro (감마선 조사 황기, 백출 및 승마 열수 추출물의 in vitro 유전독성학적 안전성 평가)

  • 박혜란;함연호;정우희;정일윤;조성기
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.31 no.5
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    • pp.910-916
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    • 2002
  • As the utilization of medicinal herbs in food and bio-industry increases, safe hygienic technologies for them are demanded. To consider the possibility of application of radiation technology for this purpose, the genotoxi-cological safety of three r -irradiated medicinal herbs were studied. Astragali Radix, Atractylodes Rhizoma and Cimicifugae Rhizoma were irradiated at 10 kGy, and then were extracted with hot water. The genotoxicity of the extracts was examined in two short-term in vitro tests: (1) Salmonella reversion assay (Ames test) in strains of TA98 and TA100; (2) Micronucleus test in cultured Chinese hamster ovary (CHO) cells. The extract was treated at maximum doses of 5 mg/plate in Salmonella reversion assay, and 1 mg/mL in micronucleus test where growth of CHO cells was inhibited by 50%. In Salmonella reversion assay with or without metabolic activation, both ex-tracts of irradiated and non-irradiated herbs showed no significant differences in formation of revertant colonies compared with the negative control. And also in micronucleus test, the incidences of micronucleus in CHO cells cultured with extracts of irradiated herbs were almost same as negative control in less than 3%. These results of two in vitro tests suggest that ${\gamma}$-irradiated herbs do not show mutagenicity and cytogenetic toxicity. Further tests of in vivo genotoxicity and chronic toxicity are needed to ascertain the safety of ${\gamma}$-irradiated herbs.