• 제목/요약/키워드: Activity assay

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Antifungal Activity of Bee Venom and Sweet Bee Venom against Clinically Isolated Candida albicans

  • Lee, Seung-Bae
    • Journal of Pharmacopuncture
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    • v.19 no.1
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    • pp.45-50
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    • 2016
  • Objectives: The purpose of this study was to investigate the antifungal effect of bee venom (BV) and sweet bee venom (SBV) against Candida albicans (C. albicans) clinical isolates. Methods: In this study, BV and SBV were examined for antifungal activities against the Korean Collection for Type Cultures (KCTC) strain and 10 clinical isolates of C. albicans. The disk diffusion method was used to measure the antifungal activity and minimum inhibitory concentration (MIC) assays were performed by using a broth microdilution method. Also, a killing curve assay was conducted to investigate the kinetics of the anti-fungal action. Results: BV and SBV showed antifungal activity against 10 clinical isolates of C. albicans that were cultured from blood and the vagina by using disk diffusion method. The MIC values obtained for clinical isolates by using the broth microdilution method varied from $62.5{\mu}g/mL$ to $125{\mu}g/mL$ for BV and from $15.63{\mu}g/mL$ to $62.5{\mu}g/mL$ for SBV. In the killing-curve assay, SBV behaved as amphotericin B, which was used as positive control, did. The antifungal efficacy of SBV was much higher than that of BV. Conclusion: BV and SBV showed antifungal activity against C. albicans clinical strains that were isolated from blood and the vagina. Especially, SBV might be a candidate for a new antifungal agent against C. albicans clinical isolates.

Antimicrobial Activity of Elfvingia applanata extract alone and in Combination with Some Antibiotics (Elfvingia applanata 엑스가 수종 항생물질의 항균력에 미치는 병용효과)

  • Kim, Young-So;Rym, Kyo-Hwan;Lee, Chong-Kil;Han, Seong-Sun
    • YAKHAK HOEJI
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    • v.38 no.6
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    • pp.742-748
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    • 1994
  • As part of our search for less toxic antimicrobial agents from natural resources, the carpophores of Elfvingia applanata$(P_{ers}.)K_{ARST}.$ was extracted with hot water. EA, the aqueous extract from the carpophores of E. applanata, was lyophilized and a dark brownish powder was obtained. Antimicrobial activity of EA was tested in vitro against Gram positive and Gram negative bacteria by serial broth dilution method, and the antimicrobial activity was expressed by minimal inhibitory concentration(MIC). Among fourteen species of bacteria tested, the antimicrobial activity of EA was the most potent against Proteus vulgaris showing MIC of 1.250 mg/ml. To investigate the effect of antimicrobial combinations of EA with four kinds of antibiotics(ampicillin, cefazolin, oxytetracycline and chloramphenicol), the fractional inhibitory concentration index(FICI) was determined by checkerboard assay for each strain. The antimicrobial combinations of EA with four kinds of antibiotics resulted in synergism in four instances, but no antagonism was observed. Four instances of synergism were observed when EA was combined with ampicillin against Micrococcus luteus, with cefazolin against Bacillus subtilis, with cefazolin against Micrococcus luteus and with oxytetracycline against Staphylococcus aureus.

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Anti-microbial Activity of Saussurea lappa C.B. Clarke Roots

  • Chang, Kyung-Mi;Choi, Soo-Im;Chung, Sophia J.;Kim, Gun-Hee
    • Preventive Nutrition and Food Science
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    • v.16 no.4
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    • pp.376-380
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    • 2011
  • We investigated the total phenolic and flavonoid contents and the antimicrobial activity of ethanol extracts obtained from Saussurea lappa C.B. Clarke. The ethanol extracts of S. lappa C.B. Clarke were fractionated with various solvents (n-hexane, chloroform, and n-butanol). The antimicrobial activity of S. lappa C.B. Clarke was examined by disc-diffusion and micro-dilution susceptibility assays with six food-borne pathogens, and compared to that of the synthetic antibiotics. It is found that the S. lappa C.B. Clarke ethanol extract and n-hexane fraction have strong activity against B. cereus and V. parahaemolyticus strains compared to ampicillin. The inhibitory concentration ($IC_{50}$) values of hexane fraction against L. monocytogenes, B. cereus, and B. subtilis were 62.5, 250 and 500 ppm, respectively. Therefore, these data suggest that S. lappa C.B. Clarke may be useful as antimicrobial agents against food-borne pathogens.

Experimental Studies on Activity of the Cultivated Mycelia of Phellinus linteus (상황(桑黃) 배양균사체의 활성에 관한 연구(I))

  • Kong, Young-Yun;Lee, Kwan-Ki;Nam, Sang-Yun;Hong, Nam-Doo
    • Korean Journal of Pharmacognosy
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    • v.22 no.4
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    • pp.233-239
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    • 1991
  • Phellinus linteus was examined for its anticancer activity using an animal model. Water extract of Phellinus linteus was prepared from artificially cultivated mycelia. Neither toxicity nor abnormal changes of hematological parameters were observed in the rat given orally with high doses of drug extract for 15 days. ICR mice were transplanted with Sarcoma-180 tumor cells intraperitoneally and drug extract was daily given to the mice from 1 day after tumer transplantation for 3 weeks. Administration of drug extract significantly prolonged the survival duration of Sarcoma 180-transplanted mice. For the better understanding of the anticancer activity, we have examined the effect of the drug extract administration on various killer cell functions, such as natural killer(NK) cells, cytotoxic T-lymphocytes (CTL) and macrophages which have been known to be main effector cells in immune responses against tumors. The results from the 4 hr $^{51}Cr-release$ assay have shown that the drug extract augments mouse NK cell activity but neither CTL nor macrophages. It is possible, then, that the anticancer activity of the Phellinus linteus may be associated with augmentation of NK cell function in the cancerated hosts.

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Biochemical Characterization of 1-Aminocyclopropane-1-Carboxylate Oxidase in Mung Bean Hypocotyls

  • Jin, Eon-Seon;Lee, Jae-Hyeok;Kim, Woo-Taek
    • BMB Reports
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    • v.31 no.1
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    • pp.70-76
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    • 1998
  • The final step in ethylene biosynthesis is catalyzed by the enzyme 1-aminocyclopropane-1-carboxylate (ACC) oxidase. ACC oxidase was extracted from mung bean hypocotyls and its biochemical characteristics were determined. In vitro ACC oxidase activity required ascorbate and $Fe^{2+}$, and was enhanced by sodium bicarbonate. Maximum specific activity (approximately 20 nl ethylene $h^{-1}$ mg $protein^{-1}$) was obtained in an assay medium containing 100 mM MOPS (pH 7.5), $25\;{\mu}M$ $FeSO_4$, 6 mM sodium ascorbate, 1 mM ACC, 5 mM sodium bicarbonate and 10% glycerol. The apparent $K_m$ for ACC was $80{\pm}3\;{\mu}M$. Pretreating mung bean hypocotyls with ethylene increased in vitro ACC oxidase activity twofold. ACC oxidase activity was strongly inhibited by metal ions such as $Co^{2+}$, $Cu^{2+}$, $Zn^{2+}$, and $Mn^{2+}$, and by salicylic acid. Inactivation of ACC oxidase by salicylic acid could be overcome by increasing the $Fe^{2+}$ concentration of the assay medium. The possible mode of inhibition of ACC oxidase activity by salicylic acid is discussed.

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Lactobacillus acidophilus Inhibits the Helicobacter pylori Adherence

  • Lee, Yeon-Hee;Shin, En-Joo;Lee, Jin-Hui;Park, Jae-Hak
    • Journal of Microbiology and Biotechnology
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    • v.9 no.6
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    • pp.794-797
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    • 1999
  • Lactobacillus acidophilus is known to have an inhibitory activity on growth of Helicobacter pylori and this activity has been attributed to lactic acid and antibacterial agents produced by Lactobacillus. Since every Lactobacilli produces lactic acid, an another factor must exist for L. acidophilus to inhibit H. pylori growth. In this work, the inhibitory activity of L. acidophilus on H. pylori adherence was studied. An immunoabsorbent assay using TLC plate was developed and used for screening the inhibitory activity of various Lactobacilli on H. pylori adherence. Glycolipid, the attachment site for H. pylori, was isolated from blood type O red blood cells and spotted on a TLC plate. The H. pylori adherence increased linearly with increasing amounts of glycolipid spotted on the TLC plate. Various L. acidophilus strains, but not L. casei, appeared to inhibit H. pylori adherence to glycolipid, and the adherence decreased linearly as the concentration of the Lactobacillus increased. The results show that the inhibitory activity of L. acidophilus on H. pylori adherence is an another factor for L. acidophilus to inhibit H. pylori growth.

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Antioxidant Activities of Aroma Extracts in Commercially Available Red Wines in Korea

  • Woo, Koan-Sik;Jeon, Geon-Uk;Yoon, Hyang-Sik;Jeong, Heon-Sang;Lee, Jun-Soo
    • Preventive Nutrition and Food Science
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    • v.13 no.3
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    • pp.237-240
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    • 2008
  • The antioxidant activities of aroma extracts from commercially available red wines in Korea were evaluated. The aroma extracts of the red wines were extracted by simultaneous steam distillation. Antioxidant activity was measured by DPPH (1,1-diphenyl-2-picrylhydrazyl) radical scavenging activity and ABTS (2,2-azino-bis-3-ethylbenzothiazoline-6-sulphonic acid) radical cation scavenging activity. DPPH radical scavenging activity of the aroma extracts in the red wines increased with increases in the amount of wine used for aroma extraction. Antioxidant activities of domestic wine 1, imported wine 7, and imported wine 12 were 97.16, 96.72 and 94.52%/20 mL wine by DPPH assay and 7.09, 8.07 and 7.28 mg ascorbic acid equivalents per mL wine by ABTS assay, respectively. This study demonstrates potent antioxidant activities of the aroma extracts of commercially available red wines in Korea.

A Continuous Spectrophotometric Assay for NADPH-cytochrome P450 Reductase Activity Using 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium Bromide

  • Yim, Sung-Kun;Yun, Chul-Ho;Ahn, Tae-Ho;Jung, Heung-Chae;Pan, Jae-Gu
    • BMB Reports
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    • v.38 no.3
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    • pp.366-369
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    • 2005
  • NADPH-cytochrome P450 reductase (CPR) transfers electrons from NADPH to cytochrome P450 and also catalyzes the one-electron reduction of many drugs and foreign compounds. Various spectrophotometric assays have been performed to examine electron-accepting properties of CPR and its ability to reduce cytochrome $b_5$, cytochrome c, and ferricyanide. In this report, reduction of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) by CPR has been assessed as a method for monitoring CPR activity. The principle advantage of this substance is that the reduction of MTT can be assayed directly in the reaction medium by a continuous spectrophotometric method. The electrons released from NADPH by CPR were transferred to MTT. MTT reduction activity was then assessed spectrophotometrically by measuring the increase of $A_{610}$. MTT reduction followed classical Michaelis-Menten kinetics ($K_m\;=\;20\;{\mu}M$, $k_{cat}\;=\;1,910\;min^{-1}$). This method offers the advantages of a commercially available substrate and short analysis time by a simple measurement of enzymatic activity of CPR.

Expression and Characterization of the Human rpS3 in a Methylotrophic Yeast Pichia pastoris

  • Kim, Joon;Lee, Jae-Yung;Jung, Sang-Oun;Youn, Bu-Hyun;Kwon, Oh-Sik
    • Journal of Microbiology
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    • v.38 no.2
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    • pp.88-92
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    • 2000
  • A human ribosomal protein S3 (rpS3), which also functions as a DNA repair enzyme(UV endonuclease III), was expressed in a methylotrophic yeast, Pichia pastoris, and biochemically characterized. UV endonuclease activity was preiously characterized, and this activity of mammalian rpS3 was found to be non-specfic upon purification and storage. Under the Pichia expression system, the subcloned cDNA of the human rpS3 gene revealed a peptide of 42 kDa by SDS-PAGE and Western blot. The secreted form of human rpS3 rendered no endonuclease activity while the intracellular form showed UV specific endonuclease activity by the nick circle assay.

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Improved immune-enhancing activity of egg white protein ovotransferrin after enzyme hydrolysis

  • Lee, Jae Hoon;Kim, Hyeon Joong;Ahn, Dong Uk;Paik, Hyun-Dong
    • Journal of Animal Science and Technology
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    • v.63 no.5
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    • pp.1159-1168
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    • 2021
  • Ovotransferrin (OTF), an egg protein known as transferrin family protein, possess strong antimicrobial and antioxidant activity. This is because OTF has two iron binding sites, so it has a strong metal chelating ability. The present study aimed to evaluate the improved immune-enhancing activities of OTF hydrolysates produced using bromelain, pancreatin, and papain. The effects of OTF hydrolysates on the production and secretion of pro-inflammatory mediators in RAW 264.7 macrophages were confirmed. The production of nitric oxide (NO) was evaluated using Griess reagent and the expression of inducible nitric oxide synthase (iNOS) were evaluated using quantitative real-time polymerase chain reaction (PCR). And the production of pro-inflammatory cytokines (tumor necrosis factor [TNF]-α and interleukin [IL]-6) and the phagocytic activity of macrophages were evaluated using an ELISA assay and neutral red uptake assay, respectively. All OTF hydrolysates enhanced NO production by increasing iNOS mRNA expression. Treating RAW 264.7 macrophages with OTF hydrolysates increased the production of pro-inflammatory cytokines and the phagocytic activity. The production of NO and pro-inflammatory cytokines induced by OTF hydrolysates was inhibited by the addition of specific mitogen-activated protein kinase (MAPK) inhibitors. In conclusion, results indicated that all OTF hydrolysates activated RAW 264.7 macrophages by activating MAPK signaling pathway.