• Title/Summary/Keyword: assay

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Inhibitory Effect of Agaricus Mixed Prescription on Metastasis and Tumor Formation

  • Chu Ji Hee;Lim Nak Cheol;Kim Sung Hoon;Ryu Ki Won
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.17 no.2
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    • pp.560-567
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    • 2003
  • Antimetastatic effects of Agaricus mixed prescription (AMP) were studied in the respect of blood-borne metastasis. For this aim, cytotoxicity against various cancer cells and normal cells, Chicken Chorioallantoic membrane (CAM) assay, cancer cell adhesion assay, platelet aggregation assay, pulmonary colonization, life span of S-180 implanted mice, and cytokine release assay were evaluated, respectively. The results were summarized as follows; AMP did not exert any cytotoxicity against all cell lines with IC50 of 25mg/ml on B16BL6. AMP disrupted formation of CAM at 1mg/ml. AMP was suppressive in adhesion assay of B16BL6. AMP also inhibited tumor induced platelet aggregation. In pulmonary colonization assay by B16BL6, the number of colonies in the lungs was significantly decreased in sample group than in control group. In animal study with S-180, the life span of AMP treated group was extended than that of control group. IL-12 was effectively increased in AMP treated group in cytokine release assay. Taken together, AMP can be possibly applied to cancer or metastasis.

Validation of Photo-comet Assay as a Model for the Prediction of Photocarcinogenicity

  • Kim, Ji-Young;Koh, Woo-Suk;Lee, Mi-Chael
    • Toxicological Research
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    • v.22 no.4
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    • pp.423-429
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    • 2006
  • Recent reports on the photocarcinogenicity and photogerotoxicity of many compounds led to an increasing awareness for the need of a standard approach to test for photogenotoxicity. The comet assay has been recently validated as a sensitive and specific test system for the quantification of DNA damage. Thus, the objectives of this study are to investigate the utility of photo-comet assay for detecting photo-mutagens, and to evaluate its ability to predict rodent photo-carcinogenicity. Photo-comet assays were performed using L5178Y $Tk^{+/-}$ mouse lymphoma cells on five test substances (8-methoxypsoralen, chlorpromazine, lomefloxacin, anthracene and retinoic acid) that demonstrated positive results in photocarcinogenicity tests. For the best discrimination between the test substance-mediated DNA damage and the undesirable DNA damage caused by direct UV absorption, a UV dose-response of the cells in the absence of the test substances was firstly fnalized. Out of 5 test substances, positive comet results were obtained for chlorpromazine, lomefloxacin, anthracene and retinoic acid while 8-methoxypsoralen found negative. An investigation into the predictive value of this photo-comet assay for determining the photocarcinogenicity showed that photo-comet assay has relatively high sensitivity. Therefore, the photo-comet assay with mammalian cells seems to be a good and sensitive predictor of the photocarcinogenic potential of new substances.

Amperometric Detection of DNA by Electroreducation of O2 in an Enzyme-Amplified Two-Component Assay

  • Yoon Chang-Jung;Kim Hyug-Han
    • Journal of the Korean Electrochemical Society
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    • v.7 no.1
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    • pp.44-48
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    • 2004
  • The two-component type enzyme amplified amperometric DNA assay is described to use an ambient $O_2$ of the substrate of the DNA labeling enzyme. Although the assay detects DNA only at > 0.5M concentration, a concentration $\~10^6$ fold higher than the sandwich-type enzyme amplified amperometric DNA assay, it can be run with an always available substrate. The assay utilizes screen-printed carbon electrodes (SPEs) which were pre-coated by a co-electrodeposited film of an electron conducting redox hydrogel and a 37-base long single-stranded DNA sequence. The DNA in the electron conducting film hybridizes and captures, when present, the 37-base long detection-DNA, which is labeled with bilirubin oxidase (BOD), an enzyme catalyzing the four-electron reduction of $O_2$ to water. Because the redox hydrogel electrically connects the BOD reaction centers to the electrode, completion of the sandwich converts the film from non-electrocatalytic to electrocatalytic for the reduction of $O_2$ to water when the electrode is poised at 200 mV vs. Ag/hgCl. The advantage or the assay over the earlier reported sandwich type enzyme amplified amperometric DNA assay, in which the amplifying enzyme was horseradish peroxidase, is that it utilizes ambient $O_2$ instead of the less stable and naturally unavailable $H_2O_2$.

Molecular Biological Study of Anti-cancer Effects of Bee Venom Aqua-acupuncture (봉독약침(蜂毒藥鍼)의 항암효과(抗癌效果)에 대한 분자생물학적(分子生物學的) 연구(硏究))

  • Park, Chan-Yol;Seo, Jung-Chul;Choi, Do-Young;Ahn, Byoung-Choul
    • Journal of Pharmacopuncture
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    • v.3 no.1
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    • pp.1-19
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    • 2000
  • To study anti-cancer effect and molecular biological mechanism of bee venom for aqua-acupuncture, the effects of bee venom on cell viability and apoptosis were analyzed using MTT assay, tryphan blue assay, $[^3H]$thymidine release assay, flow cytometric analysis, and activity of caspase-3 protease activity assay. To explore whether anti-cancer effects of bee venom are associated with the transcriptional control of gene expression, quantitative RT-PCR analysis of apoptosis-related genes was performed. The obtained results are summarized as follows: 1. The MTT assay demonstrated that cell viability was decreased by bee venom in a dose-dependant manner. 2. Significant induction of apoptosis was identified using tryphan blue assay, $[^3H]$thymidine release assay, and flow cytomet1 ric analysis of sub $G_1$ fraction. 3. In analysis of caspase-3 protease activity, the activity had increased significantly, in a dose-dependant manner. 4. Quantitative RT-PCR analysis of the apoptosis-related genes showed that Bcl-2 and Bcl-$X_L$ were down-regulated whereas Bax was up-regulated by bee venom treatment.

Antioxidant Activity and Functional Components of Corn Silk(Zea mays L.) (옥수수 수염의 항산화 활성과 기능성분 분석)

  • Ku, Kang-Mo;Kim, Soon-Kwon;Kang, Young-Hwa
    • Korean Journal of Plant Resources
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    • v.22 no.4
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    • pp.323-329
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    • 2009
  • In order to find out superior corn line which has a strong antioxidant activity in the corn silk, antioxidant compounds and antioxidant activity were measured from various inbred and F1 lines. As a result, the contents of phenolic and flavonoid compounds range from 532 mg/100 g to 3,274 mg/100 g and from 980 mg/100 g to 2,420 mg/100 g respectively. Absorbance at 517 nm for contents of anthocyanins ranges from 0.05 to 0.76. Correlation coefficients between antioxidant compounds and various antioxidant assays such as DPPH, ABTS and FRAP were analyzed. Correlation coefficients between antioxidant activity and phenolic compounds and anthocyanin were significantly high in ABTS and FRAP assays. Considering acidic extraction condition and intervention between anthocyanins and DPPH solution, ABTS assay and FRAP assay are more suitable methods to evaluate antioxidant activity of corn silks. Especially, ABTS assay is thought to be the best method among three assays because the antioxidant activity in ABTS assay showed high correlation with phenol, flavonoid and anthocyanin compounds respectively. Among the samples, S15 which showed the highest contents of total phenolic compounds and the most potent antioxidant activity in ABTS and FRAP assay will be a good source for functional material.

Risk assessment on cytotoxicity for benzimidazole fungicides (Benzimidazole계 살균제의 세포독성 평가)

  • Lee, Je-Bong;Sung, Pil-Nam;Jeong, Mi-Hye;Shin, Jin-Sup;Kang, Kyu-Young
    • The Korean Journal of Pesticide Science
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    • v.7 no.3
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    • pp.198-206
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    • 2003
  • To assess potential risk of the benzimidazole fungicides, their cytotoxicities were evaluated. Activities of LDH(Lactic dehydrogenase) in the culture fluid of CHL(chinese hamster lung) fiberoblast cell treated with 4.0, 16.0 or $32.0{\mu}g/mL$ of carbendazim for 24 hours were elevated 2.16, 2.94 and 2.64 folds compared to the control, respectively. DNA synthesis was inhibited by 45% at $2.0{\mu}g/mL$ of carbendazim. Benzimidazole fungicides showed high toxicity to cell and mitochondria of CHL cell by Giemsa and MTT (3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyltetrazolium bromide) assay. $IC_{50}$ by the Giemsa assay of thiophanate-methyl, benomyl, carbendazim and captafol were over 125, 1.2, 30.0 and $0.3{\mu}g/mL$, respectively. $IC_{50}$ by the MTT assay of thiophanate-methyl, benomyl, carbendazim and captafol were over 125, 18.7, 20.4 and $2.6{\mu}g/mL$, respectively. Inhibitory concentration of cell median proliferation by SRB (sulforhodamin B) assay for thiophanate-methyl, carbendazim, benomyl, and captafol were 17.4, 5.3, 1.5 and $0.5{\mu}g/mL$, respectively. Accordingly, benzimidazole fungicides inhibited DNA synthesis, mitochondrial function, cell proliferation and induced cell necrosis.

Chemiluminescence immunochromatographic analysis for the quantitative determination of algal toxins

  • Pyo, Dongjin;Kim, Taehoon
    • ALGAE
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    • v.28 no.3
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    • pp.289-296
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    • 2013
  • For the quantitative detection of algal toxin, microcystin, a chemiluminescence immunochromatographic assay method was developed. The developed system consists of four parts, chemiluminescence assay strip (nitrocellulose membrane), horse radish peroxidase labeled microcystin monoclonal antibodies, chemiluminescence substrate (luminol and hydrogen peroxide), and luminometer. The performance of the chemiluminescence immunochromatographic assay system was compared with high performance liquid chromatography (HPLC) detection. The detection limit of chemiluminescence immunochromatographic assay system is several orders of magnitude lower than with HPLC. The chemiluminescence immunochromatography and HPLC results correlated very well with the correlation coefficient ($r^2$) of 0.979.

Validation Process of HPLC Assay Methods of Drugs in Biological Samples (생체시료내 약물의 HPLC 분석법에 대한 유효성 검토방법)

  • Chi, Sang-Cheol;Jun, H.-Won
    • Journal of Pharmaceutical Investigation
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    • v.21 no.3
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    • pp.179-188
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    • 1991
  • An HPLC assay method of a drug to be applied to the pharmacokinetic studies of the drug should be completely validated. The validation process for an HPLC assay method in a biological sample was discussed using the data obtained from the development of HPLC method for the simultaneous quantitation of verapamil and norverapamil in human serum. The validation criteria included were specificity, linearity, accuracy, precision, sensitivity, recovery, drug stability, and ruggedness of an assay method.

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Study of Anti-Cancer Effects of Cremastrae Appendiculatae Tuber on Stomach Cancer Cells (산자고가 위암세포에 미치는 영향)

  • 김선량;류봉하;류기원;박동원
    • The Journal of Korean Medicine
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    • v.22 no.2
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    • pp.75-83
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    • 2001
  • In order to investigate anti-cancer effect of Cremastrae Appendiculatae Tuber, this experiment was performed, in vitro. The results are as follows: 1. The MIT assay demonstrated that cell viability was decreased by Cremastrae Appendiculatae Tuber without statistical significance 2. The Apoptosis assay demonstated that apoptosis was induced by Cremastrae Appendiculatae Tuber without statistical significance 3. In quantitative RT-PCR analysis, there were no remarkable changes in expression levels of apoptosis-related genes, Bcl- 2, Bax, P53.

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Bradykinin Antagonistic Activities of Antihistamine Agents Containing Piperazine Moiety (피페라진계 항히스타민제들의 브라디키닌 길항작용)

  • 정성현;이소영;윤혜숙
    • YAKHAK HOEJI
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    • v.37 no.6
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    • pp.625-630
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    • 1993
  • Among the six antihistamine agents tested in this study, homochlorcyclizine showed the highest bradykinin antagonistic activity in the receptor binding assay as well as the isolated rat ileum assay. Schild plot analysis of bradykinin-induced ileal contraction in the presence of three different concentrations of homochlorcyclizine revealed a pA$_{2}$=6.26, and a correlation coefficient of 0.984. Homochlorcyclizine of (100 $\mu{M}$ final concentration) also showed 25% antagonistic activity in the receptor binding assay.

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