• Title/Summary/Keyword: Disc assay

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Antimicrobial Activities of Licorice Extracts from Various Countries of Origin according to Extraction Conditions (원산지별 감초추출물의 추출 조건별 항균활성)

  • Bae, Jeong Yun;Jang, Ha Na;Ha, Ji Hoon;Park, Jong-Ho;Park, Jino;Park, Soo Nam
    • Microbiology and Biotechnology Letters
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    • v.42 no.4
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    • pp.361-366
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    • 2014
  • In this study, Glycyrrhiza uralensis and Glycyrrhiza glabra extracts, with their countries of origin as Korea (Jecheon), Uzbekistan and China, were prepared under various extraction conditions. There were 8 extraction conditions which the licorice were subjected to, and all conditions had different extraction solvents, temperatures and times. Antimicrobial activity on skin flora was evaluated comparatively by a disc diffusion assay, broth macrodilution assay, and kill time curve assay. Based on the antimicrobial activity of their extract confirmed by disc diffusion assay, we established optimal extraction conditions. The Korean licorice extract (85% ethanol, $40^{\circ}C$, 12 h) showed the best activity amongst the samples examined. In particular, its antimicrobial activity against Propionibacterium acnes was the highest. Minimum inhibitory concentrations (MIC) and minimum bactericidal concentrations (MBC) of the licorice extracts revealed that the Korean licorice ($156{\mu}g/ml$ and $1,250{\mu}g/ml$) had better antimicrobial activity than that of the Uzbekistani licorice ($625{\mu}g/ml$ and $2,500{\mu}g/ml$) and the Chinese licorice ($625{\mu}g/ml$ and $5,000{\mu}g/ml$). Taken together, it was shown that Korean licorice extracted in group F (85% ethanol, $40^{\circ}C$, 12 h) had the highest antimicrobial activity amongst the licorices from the other countries of origin. These results also suggest that the optimal extraction conditions are 85% ethanol, $40^{\circ}C$, 12 h, and that licorice has a potential application as a natural preservative in cosmetics products, thereby replacing synthetic preservatives.

Comparison of Antioxidant and Antimicrobial Activities of Bracken (Pteridium aquilinum Kuhn) according to Cooking Methods (조리방법에 따른 고사리의 항산화활성 및 항균활성 비교)

  • Park, Cho-Hee;Kim, Kyoung-Hee;Yook, Hong-Sun
    • The Korean Journal of Food And Nutrition
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    • v.27 no.3
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    • pp.348-357
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    • 2014
  • This study was carried out to evaluate the yield of extract, antioxidant compounds (total phenolic and total flavonoid), antioxidant (DPPH assay, ABTS assay and reducing power), and antimicrobial activities of bracken (Pteridium aquilinum Kuhn), according to cooking methods (non-blanched, blanched and seasoned). The yield of seasoned bracken extracts showed a high value of (4.59%) followed by non-blanched bracken and blanched bracken with 2.69% and 0.30%, respectively. In the total polyphenol and flavonoid contents, seasoned bracken extracts showed higher antioxidant compounds ($96.11{\pm}0.34mg\;GAE$/100 g RW, $20.90{\pm}0.mg\;CE$/100 g RW) than non-blanched and blanched. The total antioxidant activities (DPPH assay, ABTS assay and reducing power) were shown to be in the order of seasoned bracken > non-blanched bracken > blanched bracken. In the antimicrobial activities, non-blanched bracken extracts showed antimicrobial activity against B. cereus, B. subtilis, E. cloacae, E. coli, S. enterica, and P. aeruginosa except for S. aureus. The non-blanched bracken extracts (5 and 10 mg/disc) especially showed strong antimicrobial activity against P. aeruginosa ($10.00{\pm}0.71$ and $10.25{\pm}0.35mm$). The inhibition zone diameter from the extracts of blanched bracken and seasoned bracken was not detected. Many seasonings added in the process of cooking can increase the antioxidant capacities. The overall results of this study demonstrate that the cooked bracken with seasoning would be the most efficient way of ingesting antioxidant compounds.

Antimicrobial Effect of Caesalpinia sappan L. Extract on Foodborne Bacteria (식중독 세균에 대한 소목 추출물의 항균 활성)

  • Kim, Yi-Seul;Shim, Hye-Mee;Kim, Kwang-Yup
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.45 no.7
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    • pp.1026-1034
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    • 2016
  • To develop a natural antimicrobial agent, we investigated the antimicrobial activities of 13 species of edible herbal plant extracts against major Gram-positive foodborne bacteria. Among the 13 screened edible herbal plants, Caesalpinia sappan L. showed the highest antimicrobial activity. In the paper disc agar diffusion assay, Caesalpinia sappan L. extracts had strong antibacterial activities against most Gram-positive bacteria but did not have antibacterial activities against most Gram-negative bacteria. Minimum inhibitory concentrations of the ethanol extract were 0.06 mg/mL against Clostridium difficile and Listeria monocytogenes and 0.03 mg/mL against Staphylococcus aureus. Their inhibitory activities were not reduced by heat treatment or pH adjustment against C. difficile, L. monocytogenes, and S. aureus. Antimicrobial activities were higher in ethanol extract than in distilled water extract. These results support the potential use of Caesalpinia sappan L. ethanol extract as an antimicrobial agent or functional food components against Gram-positive bacteria.

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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The Relation Between Sox9, TGF-${\beta}1$, and Proteoglycan in Human Intervertebral Disc Cells

  • Lee, Yong-Jik;Kong, Min-Ho;Song, Kwan-Young;Lee, Kye-Heui;Heo, Su-Hak
    • Journal of Korean Neurosurgical Society
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    • v.43 no.3
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    • pp.149-154
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    • 2008
  • Objective: The aim of this study is to elucidate the effects of transforming growth factor-${\beta}$ (TGF-${\beta}$)1 and L-ascorbic acid on proteoglycan synthesis, and the relationship between Sox9, proteoglycan, and TGF-${\beta}1$ in intervertebral disc cells. Methods: Human intervertebral disc tissue was sequentially digested to 0.2% pronase and 0.025% collagenase in DMEM/F-12 media and extracted cells were cultured in $37^{\circ}C$, 5% $CO_2$ incubator. When intervertebral disc cells were cultured with TGF-${\beta}1$ or L-ascorbic acid, the production level of sulfated glycosaminoglycan (sGAG) was estimated by dimethyl methyleneblue (DMMB) assay. The changes of Sox9 mRNA and protein levels via TGF-${\beta}1$ were detected by RT-PCR and Western blot analysis in each. Results: The amount of sGAG was increased with the lapse of time during incubation, and sGAG content of pellet cultured cells was much larger than monolayer culture. When primary cultured intervertebral disc cells in monolayer and pellet cultures were treated by TGF-${\beta}1$ 20 ng, sGAG content of experimental group was increased significantly compared to control group in both cultures. L-Ascorbic acid of serial concentrations (50-300 ug/ml) increased sGAG content of mono layer cultured intervertebral disc cells significantly in statistics. The co-treatment of TGF-${\beta}1$ and L-ascorbic acid increased more sGAG production than respective treatment. After treating with TGF-${\beta}1$, Sox9 mRNA and protein expression rates were significantly increased in disc cells compared with the control group. Conclusion: This study suggests that TGF-${\beta}1$ would increase sulfated glycosaminoglycan (sGAG) and other proteoglycans such as versican by elevating Sox9 mRNA and protein expressions in order.

Gene Therapy Using GM-CSF Gene Transferred by a Defective Infectious Single-cycle Herpes Virus in Micro-residual Organotropic Head and Neck Squamous Cell Cancer Model (향장기성 두경부 편평세포암종의 미세잔존암 모델에서 GM-CSF 유전자를 이입시킨 제한복제성 헤르페스바이러스 벡터를 이용한 종양백신의 유전자 치료)

  • Kim Se-Heon;Choi Eun-Chang;Kim Han-Su;Chang Jung-Hyun;Kim Ji-Hoon;Kim Kwang-Moon
    • Korean Journal of Head & Neck Oncology
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    • v.19 no.1
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    • pp.25-33
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    • 2003
  • Background and Objectives: The Herpes Simplex type 2 Defective Infectious Single Cycle virus (DISC virus) is attenuated virus originally produced as viral vaccines but are also efficient gene transfer vehicle. The main goals of this study were to examine the efficiencies of the gene transfer using DISC vectors for various head and neck squamous cell carcinoma cell lines and to evaluate the efficacy of vaccination with DISC virus carrying a immunomodulatory genes (GM-CSF) as cancer therapy in a organotopic oral cavity squamous cell cancer model. Materials and Methods : We determinated the gene transfer efficiency of DISC virus by x-gal stain method and proved gene and protein expression of DISC-GMCSF transfected SCCVII cells by RT-PCR and ELISA method. Also we evaluated the ex vivo vaccination effects of SCCVII/GMCSF (DISC-GMCSF transfected SCCVII vaccine) vaccine on preventing the recurrence of micro-residual tumor. After the vaccination of SCCVII/GMCSF, specific cytotoxic T-cell responses was evaluated by CTL assay. Results: At an MOI of 10 DISC virus showed 64-88% of transfection rates in various head and neck squamous cancer cell lines. SCCVII cells transduced by DISC virus vector (MOI=10) carrying the GM-CSF gene, produced 4.5 nanogram quantities of GM-CSF per $10^6$ cells. In vivo vaccination using tumor cells transduced ex vivo with DISC-GMCSF resulted in better protection rate against subsequent tumor recurrence in organotopic oral cavity cancer model. Although tumor free survival rate was not statistically significantly increased in vaccination group (p=0.078), tumor specific cytotocic T-cell responses were significantly increased in SCCVII/GMCSF vaccination group. Conclusion: These data demonstrate that; 1) The DISC virus vector is capable of efficient gene transfer to various head and neck squamous cancer cell lines, 2) GM-CSF secreting genetically modified tumor vaccine (SCCVII/GMCSF) efficiently protected against tumor recurrence in organotopic micro-residual oral cavity cancer model and produced tumor specific cytotoxic T-cell response. DISC virus-mediated, cytokine gene transfer may prove to be useful as a clinical therapy for head and neck cancers.

The Comparison of Sponges and PLGA Scaffolds Impregnated with DBP on Growth Behaviors of Human Intervertebral Disc Cells (DBP 스폰지와 DBP/PLGA 지지체에서의 인간 디스크세포 거동분석 비교)

  • Lee, Seon-Kyoung;Hong, Hee-Kyung;Kim, Su-Jin;Kim, Yong-Ki;Song, Yi-Seul;Ha, Yoon;Lee, Dong-Won;Khang, Gil-Son
    • Polymer(Korea)
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    • v.34 no.5
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    • pp.398-404
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    • 2010
  • We fabricated sponge and poly(lactide-co-glycolide)(PLGA) scaffolds impregnated demineralized bone particle(DBP)(DBP/PLGA) and investigated proper condition to proliferation and phenotype maintenance of intervertebral disc(IVD) cells by comparison between DBP/PLGA scaffold and DBP sponge. DBP/PLGA scaffolds were prepared by solvent casting/salt leaching. Human IVD cells were seeded in scaffolds of two types. Cell viability and proliferation according to scaffolds were analyzed by WST assay and SEM. RT-PCR was assessed to measure mRNA expression of aggrecan and type II collagen of human IVD cells. In WST assay results, cell viability in scaffolds impregnated DBP/PLGA scaffold were higher than DBP sponge. We could observe that disc cell mRNA expressed better in DBP/PLGA scaffold than DBP sponge. We concluded that the using of DBP/PLGA in terms of scaffold fabrication for bio-disc with human IVD cells is helpful growth of disc cells maintenance of phenotypes.

Screening of Oriental Herbal Medicines for Antibacterial Activities

  • Bae, O-Sung;Hwang, Jae-Ock;Ahn, Duk-Kyun;Woo, Eun-Rhan;Seo, Seon-Hee;Kim, Hyoung-Ja;Park, Ho-Koon
    • Natural Product Sciences
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    • v.4 no.1
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    • pp.32-37
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    • 1998
  • The water extracts of oriental herbal medicines which have been clinically used to treat bacterial infections in Korea were screened for in vitro antibacterial activity by the paper disc assay method. Two Gram positive bacteria, Staphylococcus aureus SG511, Bacillus subtilis ATCC 6633 and two Gram negative bacteria, Escherichia coli 055, Pseudomonas aeruginosa 9027 were used as test organisms. Among 83 of the extracts tested, 25 were active against Staphylococcus aureus SG511, 9 were active against Bacillus subtilis ATCC 6633, while none showed inhibitory activity against Eschelichia coli 055 and Pseudomonas aeruginosa 9027. Among them, Hwangyonhaedoktang plus hwangyon, Chongwisan, and Ssangbaksan showed remarkably potent antibacterial activity.

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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.

Antimicrobial and Anti-inflammatory Activities of Extracts from Glycyrrhizae radix cultured with Paecilomyces japonica (감초를 이용한 눈꽃동충하초 균사체 배양 추출물의 항균 활성 및 항염증 활성 분석)

  • Kim, Eun;Kim, Meera
    • Journal of the East Asian Society of Dietary Life
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    • v.26 no.3
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    • pp.215-222
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    • 2016
  • This study was carried out to investigate the antimicrobial and anti-inflammatory activities of ethanol extract from Glycyrrhizae radix (GR) and ethanol extract from Glycyrrhizae radix cultured with Paecilomyces japonica mycelium (GRPM). Antimicrobial activity was measured by paper disc diffusion assay and minimum inhibitory concentration (MIC). Anti-inflammatory activity was evaluated by measurement of NO production in LPS-stimulated RAW264.7 cells. For the results of the paper disc diffusion assay, GRPM showed high antimicrobial activities against Bacillus cereus, Listeria monocytogenes, and Staphylococcus aureus. In addition, the MIC of GRPM (100 ppm) was lower than that of GR (200 ppm) against L. monocytogenes. When the morphology of L. monocytogenes treated with GRPM was observed using a FE-SEM, the surface of cells treated with GRPM were damaged, and some parts of the cell wall were destroyed. The inhibitory effect on NO production in LPS-stimulated RAW264.7 cells was significantly increased by GRPM treatment. In conclusion, GRPM is superior to GR in terms of antimicrobial and anti-inflammatory activities.