• Title/Summary/Keyword: biological assay

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Comparison of Biological Activities of Synurus deltoides (Aiton) Nakai Under Different Shading Conditions

  • Jiang, Yunyao;Noh, Heesum;Wang, Myeong-Hyeon
    • Korean Journal of Plant Resources
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    • v.26 no.6
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    • pp.686-694
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    • 2013
  • Shade treatment of Synurus deltoides (Aiton) Nakai was carried out with 0, 35, and 55% shading net, and samples were marked as no shade, 35% shade, and 55% shade, respectively. We examined in vitro antioxidant and anti-inflammatory capacities using a 1,1-diphenyl-2-2-pricylhydrazyl (DPPH) radical scavenging assay, a reducing power assay, a total antioxidant assay, a metal chelating assay, a superoxide radical scavenging assay, and a nitric oxide inhibition assay. As a result, no shade and 35% shade possessed higher DPPH radical scavenging activity and reducing power ability than that of 55% shade. Notably, no shade had significantly higher total phenolic and flavonoid contents than those in the other samples. No shade exhibited significantly higher total antioxidant activity than that of 35% shade and 55% shade. However, the chelating ability of 55% shade was significantly greater than that of no shade and 35% shade; 55% shade also showed significantly higher anti-inflammatory capacity than that of no shade or 35% shade.

Rapid Quantification of Topotecan in Biological Samples by Liquid Chromatography/Tandem Mass Spectrometry

  • Shin, Beom-Soo;Lee, Mann-Hyung;Yoo, Sun-Dong
    • Journal of Pharmaceutical Investigation
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    • v.39 no.5
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    • pp.367-372
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    • 2009
  • A rapid liquid chromatography/tandem mass spectrometry (LC/MS/MS) assay method was developed for the determination of topotecan levels in rat serum. The assay utilized a single liquid-liquid extraction with a mixture of ethy l acetate and acetonitrile (6:1 v/v) and isocratic elution. The multiple reaction monitoring was based on the transition of m/z 422.0$\rightarrow$376.5 for topotecan and 315.1$\rightarrow$226.6 for clomipramine (internal standard). The developed assay was validated to demonstrate the specificity, recovery, lower limit of quantification (LLOQ), accuracy and precision. The assay was linear over a concentration range from 0.5-100 ng/mL, with LLOQ being 0.5 ng/mL using a small volume of rat serum (0.1 mL). The mean intra- and inter-day assay accuracy was 87.7-111.0% and 97.8-108.3, respectively, and the mean intra- and interday precision was between 1.6-4.3% and 3.8-10.3, respectively. The developed assay was applied to a pharmacokinetic study after a bolus i.v. injection of topotecan in rats.

Thermal-and Bio-degradation of Starch-Polyethylene Films Containing High Molecular Weight Oxidized-Polyethylene

  • Kim, Mee-Ra;Pometto, Anthony-L.
    • Preventive Nutrition and Food Science
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    • v.3 no.1
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    • pp.27-35
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    • 1998
  • Starch-polyethylene films containing high molecular weight(NW) oxidized-polyethylene and prooxidant were prepared , and thermal -and bio-degradability of the films were determined. Increased levels of starch resulted in a corresponding reduction in mechanical strength of the films. However, the addition of high MW oxidized-polyethylene did not significantly reduce the percent elongation of the films. Thefilms containing high MW oxidized-polyethylene andproosicant were degreaded faster than those containing no aadditive during the heat treatment. The films lost their measureable mechanical properties when their weight-average MW(Mw) fell below 50,000. Biodegradability of the films was determined by a pure culture assay with either Streptomyces badius 252.S. setonii 75Vi2 or S. viridosporous T7A, and by an extracellulr enzyme assay using S. setonii 75vi2. The results from pure culture assay indicated that biomass accumulation on the film surface inhibited chemical and biological degradation of the films. The extracellular enzyme assay demonstrated decrease of percent elongation and increase of carbonyl index of the films. Therefore, extracellular enzyme assay could be used as a good method to evaluate biodegradability of the films.

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Validation of Analytical Methods for Plasma Total Antioxidant Capacity by Comparing with Urinary 8-Isoprostane Level

  • Lee, Sang Gil;Wang, Taoran;Vance, Terrence M.;Hurbert, Patrice;Kim, Dae-Ok;Koo, Sung I.;Chun, Ock K.
    • Journal of Microbiology and Biotechnology
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    • v.27 no.2
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    • pp.388-394
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    • 2017
  • Although several analytical methods for measuring total antioxidant capacity (TAC) have been applied to biological samples, there were often dissimilar results due to the different principles of methods applied. Thus, this study aimed to validate four conventional analytical methods for measuring plasma TAC, including the ABTS assay, DPPH assay, FRAP assay, and ORAC assay, by comparing with urinary 8-isoprostane concentration. In addition, TAC results were compared with antioxidant enzyme activities including superoxide dismutase (SOD) and glutathione peroxidase in erythrocyte, and catalase in plasma. Plasma TAC measure by ABTS assay was strongly correlated with the result by FRAP assay. Plasma TAC by FRAP and ORAC assays were negatively correlated with erythrocyte SOD activity. The agreement among the four TAC assay methods and 8-isoprostane was determined using 95% prediction limits of linear regression, expressed as the mean of 8-isoprostane ${\pm}95%$ prediction limits. The ABTS method better agreed with 8-isoprostane than the other methods, demonstrating narrow prediction of limits. Furthermore, only plasma TAC determined by the ABTS assay was inversely correlated with urinary 8-isoprostane (r = -0.35, p < 0.05). In summary, the ABTS assay would be an appropriate method to measure overall plasma antioxidant capacity and predict the body's antioxidant status.

Aspartyl aminopeptidase of Schizosaccharomyces pombe has a molecular chaperone function

  • Lee, Song-Mi;Kim, Ji-Sun;Yun, Chul-Ho;Chae, Ho-Zoon;Kim, Kang-Hwa
    • BMB Reports
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    • v.42 no.12
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    • pp.812-816
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    • 2009
  • To screen chaperone proteins from Schizosaccharomyce pombe (S. pombe), we prepared recombinant citrate synthase of the fission yeast as a substrate of anti-aggregation assay. Purified recombinant citrate synthase showed citrate synthase activity and was suitable for the substrate of chaperone assay. Several heat stable proteins including aspartyl aminopeptidase (AAP) for candidates of chaperone were screened from the supernatant fraction of heat-treated crude extract of S. pombe. The purified AAP migrated as a single band of 47 kDa on SDS-polyacrylamide gel electrophoresis. The native size of AAP was estimated as 200 kDa by a HPLC gel permeation chromatography. This enzyme can remove the aspartyl residue at N-terminus of angiotensin I. In addition, AAP showed the heat stability and protected the aggregation of citrate synthase caused by thermal denaturation. This study showed that S. pombe AAP is a moonlight protein that has aspartyl aminopeptidase and chaperone activities.

A Sphingosine Kinase-1 Inhibitor, SKI-II, Induces Growth Inhibition and Apoptosis in Human Gastric Cancer Cells

  • Li, Pei-Hua;Wu, Jin-Xia;Zheng, Jun-Nian;Pei, Dong-Sheng
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.23
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    • pp.10381-10385
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    • 2015
  • SKI-II has been reported as an inhibitor of sphingosine kinase 1 and has been extensively used to prove the involvement of sphingosine kinase and sphingosine-1-phosphate (Sphk1) in cellular processes. In the current study, we investigated the effects of SKI-II and its potential mechanisms in human gastric cancer SGC7901 cells. After treatment with SKI-II, cell growth, cell cycle distribution, apoptosis, expression of Sphk1, NF-${\kappa}B$, Bcl-2, Bax and p27 were assessed by MTT assay, flow cytometry, electron microscopy, immunocytochemistry and Western-blot assay, respectively. Our results showed that SKI-II markedly inhibited SGC7901 cell survival in a dose-dependent manner, reduced cell proliferation with accumulation of cells in the G0/G1 phase and induced apoptosis in the tumor cells. Furthermore, Western blotting and immunocytochemistry showed that the expression of p27 and Bax was increased significantly, but the expression of NF-${\kappa}B$, Bcl-2 and Sphk1 decreased by different degrees. These results indicate that SKI-II induced cell growth arrest and apoptosis. The increased apoptotic sensitivity of SGC7901 was correlated with NF-${\kappa}B$ or Bcl-2/Bax activation.

A Revised Assay for Monitoring Autophagic Flux in Arabidopsis thaliana Reveals Involvement of AUTOPHAGY-RELATED9 in Autophagy

  • Shin, Kwang Deok;Lee, Han Nim;Chung, Taijoon
    • Molecules and Cells
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    • v.37 no.5
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    • pp.399-405
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    • 2014
  • Autophagy targets cytoplasmic cargo to a lytic compartment for degradation. Autophagy-related (Atg) proteins, including the transmembrane protein Atg9, are involved in different steps of autophagy in yeast and mammalian cells. Functional classification of core Atg proteins in plants has not been clearly confirmed, partly because of the limited availability of reliable assays for monitoring autophagic flux. By using proUBQ10-GFP-ATG8a as an autophagic marker, we showed that autophagic flux is reduced but not completely compromised in Arabidopsis thaliana atg9 mutants. In contrast, we confirmed full inhibition of auto-phagic flux in atg7 and that the difference in autophagy was consistent with the differences in mutant phenotypes such as hypersensitivity to nutrient stress and selective autophagy. Autophagic flux is also reduced by an inhibitor of phosphatidylinositol kinase. Our data indicated that atg9 is phenotypically distinct from atg7 and atg2 in Arabidopsis, and we proposed that ATG9 and phosphatidylinositol kinase activity contribute to efficient autophagy in Arabidopsis.

Endophytic Bacillus subtilis MJMP2 from Kimchi inhibits Xanthomonas oryzae pv. oryzae, the pathogen of Rice bacterial blight disease

  • Cheng, Jinhua;Jaiswal, Kumar Sagar;Yang, Seung Hwan;Suh, Joo-Won
    • Journal of Applied Biological Chemistry
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    • v.59 no.2
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    • pp.149-154
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    • 2016
  • An endophytic bacterial strain was isolated from kimchi, a Korean traditional fermented Brassica campestris and identified as Bacillus subtilis MJMP2 based on the 16S rRNA sequence. This strain showed strong antagonistic activity against Xanthomonas oryzae pv. oryzae (Xoo) KACC10331, the pathogen of bacterial rice blight disease, as well as activity against some other rice phytopathogenic fungi. The active compound was purified through size-exclusion chromatography and preparative High-performance liquid chromatography. The molecular weight was determined as m/z 1043 by mass spectroscopy, which is identical to that of iturin A. Furthermore, a crude extract from the culture supernatant of Bacillus subtilis MJMP2 showed inhibitory activity against rice blight disease in both a rice leaf explant assay and a pot assay. The crude extract also enhanced the length of roots of Arabidopsis thaliana. These results suggest that the strain Bacillus subtilis MJMP2 could be used as a biological agent to control rice blight disease.