• Title/Summary/Keyword: Activity assay

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Use of a Sensitive Chemiluminescence-Based Assay to Evaluate the Metabolic Suppression Activity of Linezolid on Methicillin-Resistant Staphylococcus aureus Showing Reduced Susceptibility to Vancomycin

  • Komatsu, Mitsutakal;Tajima, Yutaka;Ito, Teruyo;Yamashiro, Yuichiro;Hiramatsu, Keiichi
    • Journal of Microbiology and Biotechnology
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    • v.19 no.7
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    • pp.734-741
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    • 2009
  • Recently, strains of methicillin-resistant Staphylococcus aureus (MRSA) with reduced susceptibility to vancomycin (VCM) have been clinically isolated. The antibacterial activity of a new drug, linezolid (LZD), in such a strain was evaluated by measuring bacterial metabolic activity. A total of 73 MRSA strains having various susceptibilities to VCM were subjected to a novel and highly sensitive chemiluminescence-based assay. LZD MIC in the tested strains, measured by the microbroth dilution method, was within the range 1-4 mg/l (mostly ${\leq}2$mg/l), except for one LZD-resistant strain (NRS127; MIC=7 mg/l), and showed no correlation with VCM resistance. The chemiluminescence assay demonstrated that bacterial metabolic activity was strongly suppressed with increasing LZD concentration. The chemiluminescence intensity curve had a low baseline activity without tailing in most strains. The present results suggest that LZD has strong antibacterial activity against MRSA strains, and would be effective for treatment of infections that are poorly responsive to VCM. The chemiluminescence assay facilitated sensitive and discriminative susceptibility testing within a relatively short time.

Comprehensive Evaluation of the Anti-Helicobacter pylori Activity of Scutellariae Radix

  • Lee, Ba Wool;Park, Il-Ho;Yim, Dongsool;Choi, Sung Sook
    • Natural Product Sciences
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    • v.23 no.1
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    • pp.46-52
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    • 2017
  • The aim of this study was to evaluate the anti-Helicobacter pylori activity of fractions and major aglycon compounds (baicalein, chrysin, oroxylin A, wogonin) of Scutellariae Radix. Minimum inhibitory concentration (MIC) measurement, DPPH radical-scavenging assay, DNA protection assay, and urease inhibition analysis were performed. The ethyl acetate (EtOAc) fraction showed the potent anti-Helicobacter activity, and therefore, compounds in the EtOAc fraction were subjected to further assay. The MICs of chrysin, oroxylin A, and wogonin against Helicobacter pylori 26695 were 6.25, 12.5 and $25{\mu}g/mL$, respectively. Baicalein exhibited the most effective DPPH radical-scavenging activity. DNA protection using Fenton reaction, chrysin, oroxylin A, and wogonin showed effective DNA protective effect. This result was also confirmed by quantitative real-time polymerase chain reaction (qRT-PCR). Regarding Jack bean urease (0.5 mg/mL, 50 unit/mg) inhibition, 20 mM ofbaicalein and chrysin inhibited urease activity by 88.2% and 72.5%, respectively.

Measurement of Antiviral Activities Using Recombinant Human Cytomegalovirus

  • Song, Byung-Hak;Lee, Gyu-Cheol;Lee, Chan-Hee
    • Journal of Microbiology
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    • v.38 no.4
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    • pp.255-259
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    • 2000
  • For rapid and sensitive measurement of antiviral activities, application of a recombinant virus containing firefly luciferase gene was attempted. Recombinant human cytomegalovirus (HCMV) containing luciferase gene driven by HCMV late gene pp28 promoter (HCMV/pp28-luc) was used to test the antiviral activities of three known compounds and the result was compared with results from the conventional plaque assay for measuring the production of infectious viruses. When human fibroblast cells were infected with HCMV/pp28-luc, luciferase activity was observed at 2 days after infection and reached maximum at 6 days after infection, whereas the production of infectious virus was maximal at 4 days after infection. The antiviral activities of ganciclovir, acyclovir, and papaverine were measured in HFF cells infected with HCMV/PP28-luc and the luciferase activity was compared with the infectious virus titers. Luciferase activity decreased as the concentration of ganciclovir or papaverine increased, while there was a slight decrease in luciferase activity with acyclovir. The level of the decrease in Luciferase activity was comparable to the level of decrease in the production of infectious virus. Therefore, the antiviral assay using recombinant virus HCMV/pp28-luc resulted in sensitivity similar to the conventional plaque assay with a significant reduction in assay time.

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Screening of Sterol Biosynthesis Inhibitors from Natural Products Using Recombinant Yeast Carrying Human Lanosterol Synthase

  • Sung, Chung-Ki;Kim, Eun-A;Chu, Yun-Ho;Shibuya, Masaaki;Ebizuka, Yutaka
    • Natural Product Sciences
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    • v.9 no.4
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    • pp.299-303
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    • 2003
  • For the screening of inhibitors of sterol biosynthesis from natural products, a simple and rapid assay method was developed using recombinant yeast carrying human lanosterol synthase, main target of this assay method. Sterol biosynthesis inhibition activity was monitored only by the inhibition of growth of the recombinant yeast. By changing the substrate, this assay method can figure out which step is inhibited in the sterol biosynthesis by the test material. With this assay method total 102 plant samples were screened for their inhibitory activity of sterol biosynthesis. Among plant water extracts screened, 11 plant samples showed inhibitory activity on sterol biosynthesis in ergosterol (-) medium. For selection of the specific inhibitory materials, 11 plant samples were reassayed in ergosterol (+) medium. After all 5 plant samples, Abutilon avicennae Gaertn. (stem), Alnus japonica Steud. (stem), Amaranthus mangostanus L. (aerial part), Philadelphus schrenckii Pupr. (leaf) and Pimpinelia brachycarpa Nakai (aerial part), showed specific inhibitory activity.

Antibacterial Activity of Cinnamaldehyde and Estragole Extracted from Plant Essential Oils against Pseudomonas syringae pv. actinidiae Causing Bacterial Canker Disease in Kiwifruit

  • Song, Yu-Rim;Choi, Min-Seon;Choi, Geun-Won;Park, Il-Kwon;Oh, Chang-Sik
    • The Plant Pathology Journal
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    • v.32 no.4
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    • pp.363-370
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    • 2016
  • Pseudomonas syringae pv. actinidiae (Psa) causes bacterial canker disease in kiwifruit. Antibacterial activity of plant essential oils (PEOs) originating from 49 plant species were tested against Psa by a vapor diffusion and a liquid culture assays. The five PEOs from Pimenta racemosa, P. dioica, Melaleuca linariifolia, M. cajuputii, and Cinnamomum cassia efficiently inhibited Psa growth by either assays. Among their major components, estragole, eugenol, and methyl eugenol showed significant antibacterial activity by only the liquid culture assay, while cinnamaldehyde exhibited antibacterial activity by both assays. The minimum inhibitory concentrations (MICs) of estragole and cinnamaldehyde by the liquid culture assay were 1,250 and 2,500 ppm, respectively. The MIC of cinnamaldehyde by the vapor diffusion assay was 5,000 ppm. Based on the formation of clear zones or the decrease of optical density caused by these compounds, they might kill the bacterial cells and this feature might be useful for managing the bacterial canker disease in kiwifruit.

Physiological Activity of Extracts of Bark from Ulmus davidiana var. japonica and its fractions (유근피 추출물 및 분획의 생리활성)

  • Yang, Sun a;Kim, A young;Pyo, Byoung sik;Kim, Sun min
    • Korean Journal of Pharmacognosy
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    • v.50 no.2
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    • pp.112-117
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    • 2019
  • In this study, extracts of bark from Ulmus davidiana var. japonica and its fractions were investigated the antioxidative, antibacterial and tyrosinase inhibition activity for physiological activity towards functional applications. In the measurement of DPPH radical scavenging activity, the ethyl acetate fraction showed higher radical scavenging ability than others. Moreover, in the tyrosinase inhibition assay, showed that the ethyl acetate fraction has good inhibition effects. Results of the DPPH radical scavenging and tyrosinase inhibition activity are related with the total polyphenol concentrations of ethyl acetate fraction. In antibacterial activity used to find out by utilizing the disc diffusion assay, chloroform fraction showed strong effect against Staphylococcus aureus and S. epidermidis. These results are related with the flavonoid contents of chloroform fraction. On the other hand, in the L929 cell viability measurement by MTT assay, the hexane, butanol and aqueous fraction treated at high concentration were showed cytotoxicity. But the others samples were exhibited a moderate viabilities. As a result of investigated the antioxidant and tyrosinase inhibition activity, the ethyl acetate fraction could be applicable for cosmetics related fields. And the chloroform fraction showed significant antibacterial activity against S. aureus and S. epidermidis.

Study on the Estrogenic Activity of Di-(2-Ethylhexyl) Adipate in E-Screen Assay and Uterotrophic Assay (E-screen assay 및 자궁비대반응시험 (Uterotrophic assay)을 이용한 di-(2-ethylhexyl) adipate의 에스트로겐성 작용에 관한 연구)

  • Han, Soon-Young;Kim, Hyung-Sik;Han, Sang-Kook;Lee, Rhee-Da;Yang, Kyu-Whan;Park, Kui-Lea
    • Korean Journal of Food Science and Technology
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    • v.32 no.4
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    • pp.964-969
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    • 2000
  • Di-(2-ethylhexyl) adipate(DEHA) has been used extensively as a plasticizer in the manufacture of plastic products such as PVC films. Though, phthalate esters plasticizers have been known to induce endocrine system-mediated responses, few studies have been conducted for the screening of estrogenic activity of DEHA, an adipate plasticizer. This study was initiated to evaluate the estrogenic activity of DEHA by in vitro E-screen assay and in vivo uterotrophic assay. MCF-7 human breast cancer cells were treated with $DEHA(5{\times}10^{-9}{\sim}5{\times}10^{-4}\;M)$, for 144 hr, and cell proliferation was determined by sulforhodamine B(SRB) assay. DEHA dissolved in corn oil was administered subcutaneously to ovariectomized(OVX) female Sprague-Dawley rats at dosage levels of 0, 2, 20 and 200 mg/kg/day for three consecutive days. Rats were sacrificed 24 hr after final treatment and vagina and uterus(wet and blotted) weights were obtained. E-screen assayed DEHA did not generate cell proliferation at treated concentrations$(5{\times}10^{-9}{\sim}5{\times}10^{-4}\;M)$, whereas 17 ${\beta}-estradiol$(E2), the positive control, induced cell proliferation at low concentrations$(5{\times}10^{-14}{\sim}5{\times}10^{-9}\;M)$. In the uterotrophic assay, DEHA did not change vagina and uterus(wet and blotted) weights at dosage levels up to 200 mg/kg/day treatment. These results demonstrated that DEHA did not exhibit the estrogenic activity as determined by in vitro E-screen assay and in vivo uterotrophic assay.

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Screening of Korean Medicinal and Food Plants with Antitumoral Activity (국내 약용 및 식용식물중 항종양활성 식물탐색)

  • Chung, Ill-Min;Kim, Kwang-Ho;Ahn, Joung-Kuk;Ahn, Jong-Seog;Ahn, Soon-Cheol
    • Korean Journal of Medicinal Crop Science
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    • v.7 no.1
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    • pp.37-44
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    • 1999
  • This study was conducted to screen antitumoral activity by in vitro bioassay method using 60 Korean medicinal and food plants extracted by 80% EtOH. Antitumor activity test was applied by the PKC(protein kinase C) and antibleb formation, PLC (Phospholipase C), and colorimetric tetrazolium assay (MTT assay) methods. Chenopodjum album and black Glycine max showed high antitumoral activity by 73.5% and 81.0%, respectively, against PKC by bleb-forming assay and PKC enzyme assay on human chronic leukemia K562 cell. Black Glycine max also showed 91.2% antitumoral activity in the PLC method and the lowest $IC_{50}$ $value(4.7{\mu}g/ml)$ by MTT method against P-338 cell line. In the effect of the concentration treatment on antitumoral test, the more concentration indicated the more activity value.

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Antioxidant Activity of Orange Flesh and Peel Extracted with Various Solvents

  • Park, Jae-Hee;Lee, Minhee;Park, Eunju
    • Preventive Nutrition and Food Science
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    • v.19 no.4
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    • pp.291-298
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    • 2014
  • The aim of this study was to investigate the antioxidant activity of orange (Citrus auranthium) flesh (OF) and peel (OP) extracted with acetone, ethanol, and methanol. Antioxidant potential was examined by measuring total phenolic content (TPC), 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity (RSA), total radical-trapping antioxidant potential (TRAP), oxygen radical absorbance capacity (ORAC), and cellular antioxidant activity (CAA). The comet assay was used to determine the protective effects of OF and OP against $H_2O_2$-induced DNA damage. TPC was highest in the acetone extracts of OF and OP. DPPH RSA was also higher in the acetone extracts than in the ethanol extracts. The DPPH RSA was highest in the acetone extracts of OF. The TRAP and ORAC values of the all extracts increased in a dose-dependent manner. In the TRAP assay, the acetone extracts of OF and OP had the lowest $IC_{50}$ values. In the CAA assay, the methanol and acetone extracts of OP had the lowest $IC_{50}$ values. All of the samples protected against $H_2O_2$-induced DNA damage in human leukocytes, as measured by the comet assay, but the acetone extracts of OP had the strongest effect. These results suggest that acetone is the best solvent for the extraction of antioxidant compounds from OF and OP. Furthermore, the high antioxidant activity of OP, which is a by-product of orange processing, suggests that it can be used in nutraceutical and functional foods.

Probiotic Properties of Lactobacillus brevis KU200019 and Synergistic Activity with Fructooligosaccharides in Antagonistic Activity against Foodborne Pathogens

  • Kariyawasam, Kariyawasam Majuwana Gamage Menaka Menike;Yang, Seo Jin;Lee, Na-Kyoung;Paik, Hyun-Dong
    • Food Science of Animal Resources
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    • v.40 no.2
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    • pp.297-310
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    • 2020
  • This study aims to evaluate the probiotic properties of Lactobacillus brevis (L. brevis) KU200019 and the synergistic activity with prebiotics on antimicrobial activity, and the potential application as an adjunct culture in fermented dairy products. The commercial strain, L. brevis ATCC 14869 was used as reference strain. L. brevis KU200019 was showed higher viability in simulated gastric (99.38±0.21%) and bile (115.10±0.13%) conditions compared to reference strain. L. brevis KU200019 exhibited antimicrobial activity against various foodborne pathogens. The supplementation of fructooligosaccharides (FOS) enhanced viability of lactic acid bacteria (>8 Log CFU/mL) and antioxidant activity [2,2-diphenyl-2-picrylhydrazyl radical assay (DPPH) assay, 31.23±1.14%; 2,2-azinobis (3-ethylbenzothiazoline-6-sulfonic acid (ABTS) assay, 38.82±1.46%] in fermented skim milk during refrigerated storage. L. brevis KU200019 was distinguished from the reference strain by its higher probiotic potential, antimicrobial activity, and higher antioxidant activity in fermented milk. Therefore, L. brevis KU200019 with FOS was demonstrated promising properties for further application in fermented dairy products with enhanced safety and quality.