• Title/Summary/Keyword: microbial assay

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Inhibitory Activity against Protein Kinase C of Some Medicinal Plants (수종 생약의 Protein kinase C 저해활성)

  • Lee, Hyun-Sun;Ahn, Soon-Cheol;Kim, Bo-Hyun;Park, Moon-Su;Oh, Won-Keun;Yoon, Byung-Dae;Ahn, Jong-Seog;Mheen, Tae-Ick
    • Korean Journal of Pharmacognosy
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    • v.23 no.3
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    • pp.142-145
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    • 1992
  • MeOH extract of twenty medicinal herbs were screened for their effects against protein kinase C (PKC) using bleb-forming assay and PKC enzyme assay. Smilax china and Sanguisorba officinalis showed potent anti-PKC activity. Campsis grandiflora and Galla Halepensis showed moderate inhibitory effect on PKC.

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Screening of New Bioactive Materials from Microbial Extracts of Soil Microorganism (I) Antimicrobial Activity from 200 Sampled Using Microdilution Assay

  • Jung, Sang-Oun;Kim, Joon;Chang, Il-Moo;Ryu, Jae-Chun
    • Archives of Pharmacal Research
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    • v.21 no.3
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    • pp.278-285
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    • 1998
  • The microdilution assay recommended by NCCLS (National Committee for Clinical Laboratory Standards) is one of the standardized methods of antibiotic susceptibility test. This method has been widely used clinically to obtain MIC values of antibiotics on pathogenic microorganisms. It is more convenient, rapid and simple to test many samples than other test methods such as agar diffusion assay and broth macrodilution assay. The screening of antimicrobial agents from microbial extracts is too laborious in its process. Therefore, a number of screening methods having more simple procedure have been developed. In our laboratory, we applied microdilution assay for screening the antimicrobial agents. This assay showed dose-response results and was more sensitive than disc diffusion assay in our system. We tested 200 samples of microbial extracts originated from 100 microbial strains and selected several samples as potential candidates. In this report, we show that the microdilution assay is more convenient method in screeing of antibiotic susceptibility than those previously reported.

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Use of cccDNA Breakage Assay and Sulforhodamine B Assay for the Prescreening of Antitumor Agents from Microbial Sources (미생물 배양액으로부터 항암제의 예비선별을 위한 cccDNA Breakage 활성검정과 Assay Sulforhodamine B 활성검정의 이용)

  • Lee, Sang-Han;Lee, Dong-Sun;Kim, Jong-Guk;Hong, Soon-Duck
    • Journal of Life Science
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    • v.8 no.1
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    • pp.67-71
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    • 1998
  • In order to develop new antitumor agents from fermentation broths, we used cccDNA breakage assay abd sulforhodamine B assay for prescreening. As a result, it was shown that sample reach 3.3% when using cccDNA breakage assay. In sulforhodamine B assay, we obtained 4 acive fraction against A549 (a cell line of human lung carcinoma) and SK-OV-3 (a cell line of human adenocarcinoma). These results suggest that these assay would be a promising method for antitumor prescreening from microbial sources.

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Evaluation of Microbial Epoxide Hydrolase Activity Based on Colorimetric Assay Using 4-(p-nitrobenzyl) Pyridine (4-(p-Nitrobenzyl)pyridine의 색깔반응을 이용한 미생물 epoxide hydrolase의 활성 평가)

  • Kim Hee Sook;Lee Eun Yeol
    • Journal of Life Science
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    • v.15 no.3 s.70
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    • pp.332-336
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    • 2005
  • Epoxide hydrolase activities of various microbial cells were analyzed by colorimetric assay based on alkylation of epoxides with 4-(p-nitrobenzyl)pyridine (NBP). The epoxide hydrolase activity was determined by measuring the decrease of color intensity at 560 nm due to the decrease of styrene oxide substrate by epoxide hydrolase-catalyzed hydrolysis reaction. The experimental conditions of NBP colorimetric assay were optimized for the efficient measurement of epoxide hydrolase activities from various microbial cells.

Application of ATP Bioluminescence Assay for a Rapid Estimation of Microbial Levels in Mackerel(Scomber japonicus) (고등어 표피의 미생물 오염도 신속측정을 위한 ATP Bioluminescence assay)

  • Oh, Se-Wook;Jo, Jin-Ho;Lee, Nam-Hyouck
    • Korean Journal of Food Science and Technology
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    • v.31 no.5
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    • pp.1345-1348
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    • 1999
  • The utility of a bioluminescence adenosine triphosphate(ATP) assay method for estimating bacterial levels in mackerel(Scomber japonicus) was investigated. Mackerel was stored at $1^{\circ}C$ throughout 10 days and its RLU(relative light unit) and APC(aerobic plate count) was determined. The ATP bioluminescence assay was validated during the storage of 32 samples, resulting in an agreement between the ATP assay and standard plate count methods of over 90% credibility. Therefore, ATP bioluminescence assay was considered as a rapid and near real-time means in estimating the microbial load on mackerel skin.

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Determination of Microbial Growth by Protein Assay in an Air-Cathode Single Chamber Microbial Fuel Cell

  • Li, Na;Kakarla, Ramesh;Moon, Jung Mi;Min, Booki
    • Journal of Microbiology and Biotechnology
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    • v.25 no.7
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    • pp.1114-1118
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    • 2015
  • Microbial fuel cells (MFCs) have gathered attention as a novel bioenergy technology to simultaneously treat wastewater with less sludge production than the conventional activated sludge system. In two different operations of the MFC and aerobic process, microbial growth was determined by the protein assay method and their biomass yields using real wastewater were compared. The biomass yield on the anode electrode of the MFC was 0.02 g-COD-cell/gCOD-substrate and the anolyte planktonic biomass was 0.14 g-COD-cell/g-COD-substrate. An MFC without anode electrode resulted in the biomass yield of 0.07 ± 0.03 g-COD-cell/g-CODsubstrate, suggesting that oxygen diffusion from the cathode possibly supported the microbial growth. In a comparative test, the biomass yield under aerobic environment was 0.46 ± 0.07 g-COD-cell/g-COD-substrate, which was about 3 times higher than the total biomass value in the MFC operation.

A Study on the Microbiological Assay of Bacampicillin Hydrochloride (염산바캄피실린의 역가 검정법에 관한 연구)

  • Chung, Kyeong-Soo;Kim, Young-Jun;Lee, Song-Ae;Kim, Byung-Kak
    • The Korean Journal of Mycology
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    • v.8 no.1
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    • pp.69-71
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    • 1980
  • To establish a competent method of microbiological assay for bacampicillin, one of new semi-synthetic derivatives of ${\beta}-lactam$ antibiotics, a comparison was carried out between different conditions for hydrolysis of bacampicillin into ampicillin which was then subjected to cylinder plate method. The results showed that the use of carboxylic ester hydrolase in vitro as a pretreatment of it in either pH value 6 or 8 was feasible and that the cylinder plate method with Sarcina lutea was adequate for potency estimation.

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Rapid Detection of Noroviruses in Fecal Samples and Shellfish by Nucleic Acid Sequence-based Amplification

  • Kou Xiaoxia;Wu Qingping;Zhang Jumei;Fan Hongying
    • Journal of Microbiology
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    • v.44 no.4
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    • pp.403-408
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    • 2006
  • The purpose of this study was to determine the efficacy of a nucleic acid sequence-based amplification (NASBA) method of detecting noroviruses in artificially and naturally contaminated shellfish. We used 58 fecal samples that tested positive for noroviruses with electron microscopy (EM) to develop an NASBA assay for these viruses. Oligonucleotide primers targeting the polymerase coding region were used to amplify the viral RNA in an isothermal process that resulted in the accumulation of RNA amplicons. These amplicons were detected by hybridization with digoxigenin-labeled oligonucleotide probes that were highly specific for genogroup I (GI) and genogroup II (GII) of noroviruses. The expected band of 327bp appeared in denaturing agarose gel without any nonspecific band. The specific signal for each amplicon was obtained through Northern blotting in many repeats. All fecal samples of which 46(79.3%) belonged to GII and 12(20.6%) belonged to GI were positive for noroviruses by EM and by NASBA. Target RNA concentrations as low as 5pg/ml were detected in fecal specimens using NASBA. When the assay was applied to artificially contaminated shellfish, the sensitivity to nucleic acid was 100pg/1.5g shellfish tissue. The potential use of this assay was also confirmed in naturally contaminated shellfish collected from different ponds in Guangzhou city of China, of which 24 (18.76%) out of 128 samples were positive for noroviruses; of these, 19 (79.6%) belonged to GII and 5 (20.4%) belonged to GI. The NASBA assay provided a more rapid and efficient way of detecting noroviruses in fecal samples and demonstrated its potential for detecting noroviruses in food and environmental samples with high specificity and sensitivity.

Quantitative Polymerase Chain Reaction for Microbial Growth Kinetics of Mixed Culture System

  • Cotto, Ada;Looper, Jessica K.;Mota, Linda C.;Son, Ahjeong
    • Journal of Microbiology and Biotechnology
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    • v.25 no.11
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    • pp.1928-1935
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    • 2015
  • Microbial growth kinetics is often used to optimize environmental processes owing to its relation to the breakdown of substrate (contaminants). However, the quantification of bacterial populations in the environment is difficult owing to the challenges of monitoring a specific bacterial population within a diverse microbial community. Conventional methods are unable to detect and quantify the growth of individual strains separately in the mixed culture reactor. This work describes a novel quantitative PCR (qPCR)-based genomic approach to quantify each species in mixed culture and interpret its growth kinetics in the mixed system. Batch experiments were performed for both single and dual cultures of Pseudomonas putida and Escherichia coli K12 to obtain Monod kinetic parameters (μmax and Ks). The growth curves and kinetics obtained by conventional methods (i.e., dry weight measurement and absorbance reading) were compared with that obtained by qPCR assay. We anticipate that the adoption of this qPCR-based genomic assay can contribute significantly to traditional microbial kinetics, modeling practice, and the operation of bioreactors, where handling of complex mixed cultures is required.