• Title/Summary/Keyword: NR assay

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A STUDY ON THE CYTOTOXIC EFFECTS OF MITOMYCIN C AND 5-FLUOROURACIL IN CULTURED RAT FIBROBLASTS

  • C. S. M;Park, Hong-Seog;Chung, Yeun-Tai
    • Toxicological Research
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    • v.7 no.1
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    • pp.13-20
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    • 1991
  • To investigate the cytotoxicity and genotoxicity of the DNA alkylating agnet, mitomycin C and the antimetabolite, 5-Fluorouracil (5-FU) in cultured rat fibroblasts, the colorimetric assay of netural red (NR) for cytotoxicity and for genotoxicity, sister chromatid exchange (SCE) assay and the measurement of the rate of DNA synthesis were performed in cells cultured in media containing various concentrations of mitomycin C and 5-FU. The uptake ability of neutral red decreased does-dependently. NR90 and NR50 values of mitomycin C were 1.49 nM and 6.87mM and 5-FU were 38.4mM AND 284.4Mm respectively.

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Effects of Baepungtang water extract on Cultured Spinal Sensory neurons Damaged by Xanthine Oxidase/Hypoxanthine (배풍탕(排風湯) 전탕액(煎湯液)이 XO/HX에 의해 손상(損傷)된 배양(培養) 척수감각신경세포(脊髓感覺神經細胞)에 미치는 효과(效果))

  • Yu Jin-Deok;Yun Yong-Gap
    • Herbal Formula Science
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    • v.8 no.1
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    • pp.319-328
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    • 2000
  • To evaluate the effect of Baepungtang(BPT) water extract on cultured mouse spinal sensory neuron which was inhibited by xanthine oxidase(XO) and hypoxanthine(HX)-induced oxigen radicals, MTT assay, NR assay, Neurofilament enzymeimmuno assay and LDH activity assay were carried out after the cultured mouse spinal sensory neuron were preincubated with various concentrations of BPT water extract for 3 hours prior to exposure of XO/HX. The results obtained were as follows: 1. XO/HX, a oxigen radical, decreased the survival rate of the cultured mouse spinal sensory neuron cell on NR assay and MTT assay. 2. $MTT_{50}$ value and $NR_{50}$ value of XO/HX were 30 mU/ml XO/O.2 mM HX. 3. BPT water extract have efficacy of increasing neurofilament. 4. BPT water extract have efficacy of increasing LDH activity. From above the results, It is concluded that BPT has marked efficacy as a treatment for the damages caused in the XO/HX-mediated oxidative process.

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Application of Neutral Red Staining Method to Distinguishing Live and Dead Marine Plankton for the Investigation of Efficacy of Ship's Ballast Water Treatment System (선박평형수 처리 시스템 효율 검증을 위한 해양 플랑크톤 생사판별시 Neutral red 염색법 적용 가능성 연구)

  • Hyun, Bonggil;Shin, Kyoungsoon;Chung, Hansik;Choi, Seo-Yeol;Jang, Min-Chul;Lee, Woo-Jin;Choi, Keun-Hyung
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.19 no.4
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    • pp.223-231
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    • 2014
  • In order to prevent the spread of non-indigenous aquatic species through the ballast water in commercial ships, International Maritime Organization (IMO) adopted in 2004 the International Convention for Control and Management of Ship's Ballast Water and Sediments. The Convention mandates treatment of ballast water for most transoceanic voyages and its confirmation of treatment is made with plankton live/dead assay. Fluorescein diacetate assay (FDA), which produces bright green light for live phytoplankton, has been a de facto standard method to determine the survival of marine plankton, but its staining efficacy has been in dispute. In the present study, we examined the limitation of FDA, and compared its efficacy with Neutral red (NR) staining, another promising assay and widely used especially for zooplankton mortality. For all phytoplankton species studied in the present study, except Ditylum brightwellii, the staining efficiency was <50% with FDA. The green FDA fluorescence interfered with phytoplankton autofluorescence in most samples. In contrast, NR assay stained over 90% of both phytoplankton and zooplankton species tested in this study. FDA assay also showed that green FDA fluorescence rapidly faded when phytoplankton cells were exposed to microscope light. Both FDA and NR assay were negative on formalin-killed individuals of both phytoplankton and zooplankton species. Our results suggest that NR assay is more effective for determining the survival of marine plankton and can be applied to test the efficacy of ballast water treatment.

A Study on the Cytotoxic Effects of Several Plant Extracts on the Cell viability and Cell Adhesion Activity in Cultured NIH3T3 Fibroblast (몇 가지 식물추출물이 배양 NIH3T3 섬유모세포의 세포생존율과 세포부착률에 미치는 세포독성에 관한 연구)

  • Rim, Yo-Sup;Song, Won-Seob;Seo, Young-Mi;Park, Seung-Taeck;Kim, Shin-Moo
    • Korean Journal of Clinical Laboratory Science
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    • v.42 no.3
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    • pp.116-124
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    • 2010
  • This study was aimed to clerify the cytotoxicity of some plant extracts such as Hosta longissima HONDA (HL), Hemerocallis fulva var. Kwanso REGL (HFVK), Hemerocallis fulva L (HF), Macrocapium officinale NAKAI (MO) and Mentha canadensis var. piperascens HARA (MCVP), the cultured NIH3T3 fibroblasts were treated with 25, 50, 100, 150 and $200{\mu}g/mL$ of five kinds of plant extracts for 48 hours, respectively. The cytotoxicity of plant extracts was measured by MTT and NR assays for the cell viability, and XTT assay for the cell adhesion activity. In this study, HL, MO and FHVK extracts showed the range of midtoxic-non toxic by the criteria of chemical cytotoxicity. While, the HF and MCVP extracts showed midtoxic. In the extract cytotoxicity, HL, MO and FHVK extracts showed non-toxic by the criteria of extract cytotoxicity. While, HF extract was determined as lower-toxic. In the responsive sensitivity of each plant extract on colorimetric assays, HF extract was sensitive to mitochondrial enzyme by MTT assay, lysosomal enzyme by NR assay and mitochondrial nucleus by XTT assay. While, MCVP extract was sensitive to mitochondrial enzyme by MTT assay and lysosomal enzyme by NR assay than other assays. While, HL, HFVK and MO extracts were most sensitive to NR assay. Cell culture is one of useful materials in the screening of cytotoxic and recovary effect on the putative chemical agents or plant extract. And also, colorimetric assay is regarded as very useful tools for quantitative measurement of cytotoxic effect on plant extracts in vitro.

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Effects of Yuldahansotang water extract on Cultured Primary Hippocampal Cell Culture Damaged by Hydrogen Peroxide (열다한소탕(熱多寒少湯) 전탕액(煎湯液)이 Hydrogen Peroxide에 의해 손상(損傷)된 배양(培養) 해마신경세포(海馬神經細胞)에 미치는 영향)

  • Park, Hye-Sun;Kim, Kyung-Yo;Go, Gi-Deok;Kim, Il-Hwan;Lee, Jae-Heung
    • Journal of Sasang Constitutional Medicine
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    • v.14 no.1
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    • pp.79-89
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    • 2002
  • To evaluate the effect of Yuldahansotang(YHT) water extract on cultured hippocampal cell was inhibited by hydrogen peroxide, MTT assay, NR assay, Neurofilament enzymeimmuno assay and DNA synthesis assay were carried out after the cultured hippocampal cells were preincubated with various concentrations of YHT water extract for 3 hours prior to exposure of hydrogen peroxide. The results obtained were as follows: 1. Hydrogen Peroxide decreased the survival rate of the cultured hippocampal cells on NR assay and MIT assay. 2. YHT water extract have efficacy of increasing a amount of neurofilament decreased by hydrogen peroxide in cultured hippocampal cells. 3. YHT water extract have efficacy of increasing DNA synthesis decreased by hydrogen peroxide in cultured hippocampal cells. From above the results, It is concluded that YHT has marked efficacy in preventing for the damages by hydrogen peroxide.

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Effects of Yuldahansotang water extract on Cultured Spinal Sensory Neurons Damaged by Xanthine Oxidase/Hypoxanthine (열다한소탕(熱多寒少湯) 전탕액(煎湯液)이 XO/HX에 의해 손상(損傷)된 배양척수감각신경세포(培養脊髓感覺神經細胞) 미치는 효과(效果))

  • Hong, Jeong-a;Kim, Kyung-yo;Yu, Do-gon;Park, Hye-sun;Kim, Hyung-soon
    • Journal of Sasang Constitutional Medicine
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    • v.13 no.1
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    • pp.88-96
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    • 2001
  • To evaluate the effect of Yuldahansotang(YHT) water extract on cultuted mouse spinal sensory neuron which was inhibited by xanthine oxidase(XO) and hypoxanthine(HX)-induced oxigen radicals, MIT assay, NR assay, Neurofilament enzymeimmuno assay and LDH activity assay were carried our after the cultured mouse spinal sensory neuron were preincubated with various concentrations of YHT water extract for 3 hours prior to exposure of XO/HX. The results obtained were as follows: 1. XO/HX, a oxigen radical, decreased the survival rate of the cultured mouse spinal sensory neuron cells on NR assay and MTT assay. 2. MTT50 value and NR50 value pf XO/HX were 20 mU/ml XO/0.2 mM HX and 40 mU/ml XO/0.2 mM HX. 3. YHT water extract have efficacy of increasing neurofilament. 4. YHT water extract have efficacy of increasing LDH activity. From above the results, It is concluded that YHT has marked efficacy as a treatment for the damages caused in the XO/HX-mediated oxidative process.

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Effects of Jangwon-hwan(Zhuangyuan-wan) Water Extract against Xanthine Oxidase / Hypoxanthine-induced Neurotoxicity in the Cultured Mouse Cerebral Cortical Neurons (장원환이 XO/HX에 의해 손상된 대뇌피질 신경세포에 미치는 영향)

  • 김영수;권강범;민영기;조현익;박준배;이호섭;류도곤
    • The Journal of Korean Medicine
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    • v.20 no.4
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    • pp.3-10
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    • 2000
  • In order to elucidate the toxic mechanism of neurotoxical damage and neuroprotective effect of Jangwon-hwan(Zhuangyuan-wan) water extract, this experiment was performed. Neurotoxic effects of xanthine oxidase/hypoxanthine(XO/HX) were examined by MTT and NR assay, neuroprotective effects of Jangwon-hwan(Zhuangyuan-wan) water extract were examined by neurofilament enzymeimmuno assay(EIA). XO/HX induced an increase in cell viability, and a decrease in the amount of neurofilament on cultured mouse cerebral cortical neurons in dose-dependent manner. In neuroprotective effect of herb medicine, Jangwon-hwan(Zhuangyuan-wan) water extract increased the amount of neurofilament on cultured mouse cerebral cortical neurons damaged by XO/HX. From the results, it is suggested that XO/HX showed toxic effect in cultured mouse cerebral cortical Neurons and Jangwon-hwan(Zhuangyuan-wan) water extract is very effective in the prevention of neurotoxicity induced by XO/HX.

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Development of Anticancer Agents from Korean Medicinal Plants. Part 6. -Cytotoxic Activity of the Ethyl Acetate Soluble Fraction of Lonicerae flos against Human Oral Epitheloid Carcinoma Cells- (한국산 생약으로부터 항암물질의 개발(제6보). -금은화 Ethyl Acetate 가용성 분획의 인체 구강유상피암종세포에 미치는 세포독성작용-)

  • Han, Du-Seok;Baek, Kyong-Hyun;Kim, Young-Ok;Choi, Kyu-Eun;Kwag, Jung-Suk;Baek, Seung-Hwa
    • Korean Journal of Pharmacognosy
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    • v.29 no.1
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    • pp.22-27
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    • 1998
  • This study was carried out to develop antitumor agents based on effects of the ethyl acetate soluble fraction of the methanolic extract of Lonicerae flos on human oral epitheloid carcinoma cells. Human oral epitheloid carcinoma cells were cultured in RPMI-1640 media containing 10% fetal bovine serum, antibiotic, and fungizone. After incubation for 24 hrs, the cells were treated with A, B, C, D, and E fractions for 48hrs under the same condition. MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazoliumbromide), NR (Neutral red) and SRB (Sulforhodamine B protein) assay were performed. The light microscopic findings were observed by inverted microscope. In MTT assay, fraction B was shown significant antitumor activity (P<0.001), fraction E was shown significant antitumor activities (P<0.05), but the other fractions were not shown. In NR assay, fraction B was shown significant antitumor activity (P<0.001). In SRB assay, fractions B was shown significant antitumor activities (P<0.01). fractions A and D were shown significant antitumor activities (P<0.05). but the other fractions were not shown. In light microscopy. the fraction B of the ethyl acetate soluble fraction of the methanolic extract of Lonicerae flos showed the highest antitumor activity. These finding suggested that fraction B possessed the most antitumorous agent.

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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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Development of Anticancer Agents from Korean Medicinal Plants (Part 8). - Cytotoxic Activity of Taraxaci Herba Extract against Human Skin Melanoma Cells - (한국산 생약으로부터 항암물질의 개발(제 8보) - 포공령 추출물이 인체 피부흑색종세포에 미치는 세포독성작용 -)

  • Oh, In-Kio;Yoo, Eun-Ah;Han, Du-Seok;Kang, Kil-Ung;Baek, Seung-Hwa
    • Korean Journal of Pharmacognosy
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    • v.29 no.3
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    • pp.198-203
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    • 1998
  • In the present study, we have evaluated cytotoxic effects of Taraxaci herba extract on human skin melanoma cells. The light microscopic study showed morphological changes of the treated cells. Disruptions in cell organelles were determined by calorimetric methods: MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazoliumbromide), NR (Neutral red) and SRB (Sulforhodamine B protein) assay. These results suggest that Taraxaci herba retains a potential antitumor activity.

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