• Title/Summary/Keyword: Minimal inhibition concentration

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In-Vitro, Anti-Bacterial Activities of Aqueous Extracts of Acacia catechu (L.F.)Willd, Castanea sativa, Ephedra sinica stapf and shilajita mumiyo Against Gram Positive and Gram Negative Bacteria

  • Dashtdar, Mehrab;Dashtdar, Mohammad Reza;Dashtdar, Babak;shirazi, Mohammad khabaz;Khan, Saeed Ahmad
    • Journal of Pharmacopuncture
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    • v.16 no.2
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    • pp.15-22
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    • 2013
  • Objective: Evaluations of the in-vitro anti-bacterial activities of aqueous extracts of Acacia catechu (L.F.)Willd, Castanea sativa, Ephedra sinica stapf and Shilajita mumiyo against gram-positive bacteria (Staphylococcus aureus, Streptococcus pneumonia) and gram-negative bacteria (Escherichia coli, klebsiella pneumoniae, Proteus mirabilis, Pseudomonas aeruginosa) are reasonable since these ethnomedicinal plants have been used in Persian folk medicine for treating skin diseases, venereal diseases, respiratory problems and nervous disorders for ages. Methods: The well diffusion method (KB testing) with a concentration of $250{\mu}g/disc$ was used for evaluating the minimal inhibitory concentrations (MIC). Maximum synergistic effects of different combinations of components were also observed. Results: A particular combination of Acacia catechu (L.F.) Willd, Castanea sativa, Ephedra sinica stapf and shilajita mumiyo extracts possesses an outstanding anti-bacterial activity. It's inhibiting effect on microorganisms is significant when compared to the control group (P<0.05). Staphylococcus aureus was the most sensitive microorganism. The highest anti-bacterial activity against gram-positive bacteria (Staphylococcus aureus, Streptococcus pneumonia) or gram-negative bacteria (Escherichia coli, Klebsiella pneumonia, Proteus mirabilis, and Pseudomonas aeruginosa) was exerted by formula number 2 (table 1). Conclusion: The results reveal the presence of anti-bacterial activities of Acacia catechu, Castanea sativa husk, Ephedra sp. and Mumiyo against gram-positive and gram-negative bacteria. Synergistic effects in a combined formula, especially in formula number 2 (ASLAN$^{(R)}$) can lead to potential sources of new antiseptic agents for treatment of acute or chronic skin ulcers. These results considering the significant anti-bacterial effect of the present formulation, support ethnopharmacological uses against diarrheal and venereal diseases and demonstrate use of these plants to treat infectious diseases.

Inhibition of Tyrosine Hydroxylase by $(1R,9S)-{\beta}-Hydrastine$ Hydrochloride in PC12 cells

  • Yin, Shou-Yu;Kim, Yu-Mi;Lee, Jae-Joon;Jin, Chun-Mei;Yang, Yoo-Jung;Lim, Kyo-Whan;Kang, Min-Hee;Lee, Myung-Koo
    • Natural Product Sciences
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    • v.10 no.3
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    • pp.114-118
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    • 2004
  • It is reported that $(1R,9S)-{\beta}-Hydrastine$ hydrochloride (BHSH) decreased the intracellular dopamine content by inhibiting tyrosine hydroxylase (TH) activity in PC12 cells. In this study, the inhibitory mechanisms on TH activity by BHSH in PC12 cells were investigated. BHSH treatment caused a reduction of TH activity and TH mRNA level in a dose-dependent manner. After the treatment of $20\;{\mu}M$ BHSH, TH activity and TH mRNA content were reduced at 15 min, reached the minimal levels at 6-24 h, and then recovered gradually to the control level. BHSH at $10-50\;{\mu}M$ caused a decrease in the basal intracellular cyclic AMP levels at 10 min in a concentration-dependent manner. In addition, BHSH at $20-100\;{\mu}M$ decreased the basal intracellular $Ca^{2+}$ concentration $([Ca^{2+}]_i)$ immediately in a dose-dependent manner. BHSH also inhibited the 56 mM $K^+ $ depolarization-induced elevation in $[Ca^{2+}]_i$, and blocked caffeine-activated store-operated $Ca^{2+}$ entry in PC12 cells. These data suggest that BHSH inhibits TH activity and TH gene expression, in part, through reducing cyclic AMP content and basal $[Ca^{2+}]_i$ in PC12 cells.

Two Flavonoid-Based Compounds from Murraya paniculata as Novel Human Carbonic Anhydrase Isozyme II Inhibitors Detected by a Resazurin Yeast-Based Assay

  • Sangkaew, Anyaporn;Samritsakulchai, Nawara;Sanachai, Kamonpan;Rungrotmongkol, Thanyada;Chavasiri, Warinthorn;Yompakdee, Chulee
    • Journal of Microbiology and Biotechnology
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    • v.30 no.4
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    • pp.552-560
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    • 2020
  • Human carbonic anhydrase (CA) isozyme II has been used as protein target for disorder treatment including glaucoma. Current clinically used sulfonamide-based CA inhibitors can induce side effects, and so alternatives are required. This study aimed to investigate a natural CA inhibitor from Murraya paniculata. The previously developed yeast-based assay was used to screen 14 compounds isolated from M. paniculata and identified by NMR analysis for anti-human CA isozyme II (hCAII) activity. Cytotoxicity of the compounds was also tested using the same yeast-based assay but in a different cultivation condition. Two flavonoid candidate compounds, 5, 6, 7, 8, 3', 4', 5'-heptamethoxyflavone (4) and 3, 5, 7, 8, 3', 4', 5'-heptamethoxyflavone (9), showed potent inhibitory activity against hCAII with a minimal effective concentration of 10.8 and 21.5 μM, respectively, while they both exhibited no cytotoxic effect, even at the highest concentration tested (170 μM). The results from an in vitro esterase assay of the two candidates confirmed their hCAII inhibitory activity with IC50 values of 24.0 and 34.3 μM, respectively. To investigate the potential inhibition mechanism of compound 4, in silico molecular docking was performed using the FlexX and SwissDock software. This revealed that compound 4 coordinated with the Zn2+ ion in the hCAII active site through its methoxy oxygen at a distance of 1.60 Å (FlexX) or 2.29 Å (SwissDock). The interaction energy of compound 4 with hCAII was -13.36 kcal/mol. Thus, compound 4 is a potent novel flavonoid-based hCAII inhibitor and may be useful for further anti-CAII design and development.

Antifungal and Antioxidative Activities of Yucca smallina Fern

  • Jin, Yu-Lan;Jung, Woo-Jin;Kuk, Ju-Hee;Kim, Jung-Bong;Kim, Kil-Yong;Park, Ro-Dong
    • Journal of Applied Biological Chemistry
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    • v.49 no.4
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    • pp.165-170
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    • 2006
  • The antifungal activity of crude methanolic extract and fractions from Yucca smalliana Fern. leaves, roots and flowers were investigated in vitro against a panel of plant pathogenic fungi. The minimal inhibitory concentration(MIC) was determined by an agar dilution method. Preliminary liquid culture and agar plate assays showed that the growth of Fu sarium oxysporum, Phytophthora capsici, Rhizoctonia solani and Botrytis cinerea were inhibited by Y. smalliana extracts. The extracts from flowers and leaves showed antifungal activity of 64.0% and 34.0% against F. oxysporum, 66.0% and 62.0% against P. capsici, and 27.0% and 41.0% against B. cinerea, respectively. The methanolic extract from Y. smallina leaves in distilled water was fractionated using solvents of increasing polarity: hexane, ethyl acetate and butanol. These fractions had a broad spectrum of antifungal activity, found to reside entirely in the butanol and aqueous fraction. The aqueous fraction showed inhibition rate of 60.0, 67.8, 84.6 and 58.3% against F. oxysporum, R. solani, C. gloeosporioides, and B. cinerea, respectively, and the butganol fracgtion showed 36.0, 46.0, 66.1 and 58.3%, respectively. Phenolics(e.g. flavonoids, steroids and terpenoids) were observed in the thin layer profile of the different fractions. Leave extract showed a prominent antioxidant activity totally scavenging the free radical of DPPH at a concentration of 1 mg/ml.

Studies on the cytotoxicity and Antimicrobial Effects of the Extract of Houttuynia cordata (IV) (어성초 추출물의 세포독성과 향균효과 (IV))

  • Lee Jeong-Ho;Park Nang-Kyu;Yang Eun-Yeong;Lee Hyun-Ok;Han Dong-Min;Baek Seung-Hwa
    • Journal of Society of Preventive Korean Medicine
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    • v.4 no.1
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    • pp.144-151
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    • 2000
  • This study was carried out to evaluate cytotoxic effects of Houttuynia cordata Thunberg extracts on murine leukemia tumor cell lines. Disruptions in cell organelles were determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazol iumbromide (MMT) assay. The comparison of $IC_{50}$ values of Houttuynia cordata Thunberg extracts on $L1210,\;P388D_1$ and Vero cell lines showed that the methanol extract of Houttuynia cordata Thunberg indicated the most antitumor activity in the MTT assay. In order to develop a antimicrobial agent, dried Houttuynia cordata Thunberg was extracted with several solvents, and then antimicrobial activity was investigated. The minimal inhibitory concentration (MIC) of the extracted substance against microorganisms were also examined. Antimicrobial activity of amocla and ketoconazole as references was compared to those of other solvent extracts such as $H_2O$, n-hexane, chloroform, ethyl acetate ethanol and methanol. The antimicrobial activity of all extracts from the sample had growth inhibition activity against gram-negative bacteria, yam-positive bacteria and fungi $(MIC,\;>\;200\;{\mu} g/ml)$. These results suggest that the methanol soluble extract of Houttuynia cordata Thunberg may be a valuable choice for the studies on the treaeent of murine leukemia tumor cell lines and antimicrobial agents.

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Antibacterial Effect of Ipyo-san against Methicillin-Resistant Staphylococcus aureus (입효산(立效散)의 Methicillin-Resistant Staphylococcus aureus에 대한 항균활성에 관한 연구)

  • Yoon, Jae-Hong;Choi, Yeun-Ju;Jeong, Seung-Hyun;Shin, Gil-Cho
    • The Journal of Internal Korean Medicine
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    • v.34 no.3
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    • pp.278-288
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    • 2013
  • Objectives : Methicillin-resistant Staphylococcus aureus (MRSA) has a cephalosporin and beta-lactam antibiotic-resistant strains. MRSA is one of the major pathogens causing hospital infection and the isolation ratio of MRSA has gradually increased. Consequently, increased resistance to antibiotics is causing serious problems in the world. Therefore, there is a need to develop alternative antimicrobial drugs for the treatment of infectious diseases. Methods : The antibacterial activities of Ipyo-san were evaluated against 2 strains of MRSA and 1 standard Methicillin-susceptible staphylococcus aureus (MSSA) strain by using the disc diffusion method, minimal inhibitory concentrations (MIC) assay, colorimetric assay using MTT test, checkerboard dilution test and time-kill assay performed under dark. Results : The MIC of Ipyo-san water extract against S. aureus strains ranged from 1000 to $2,000{\mu}g/ml$, so we confirmed that it had a strong antibacterial effect. Also, the combinations of Ipyo-san water extract and conventional antibiotics exhibited improved inhibition of MRSA with synergy effect. We suggest that Ipyo-san water extract against MRSA has antibacterial activity so it has potential as alternatives to antibiotic agents. For the combination test, we used Triton X-100 (TX) and DCCD for measurement of membrane permeability and inhibitor of ATPase. As a result, antimicrobial activity of Ipyo-san water extract was affected by the cell membrane. Conclusions : We suggest that the Ipyo-san water extract lead the treatment of bacterial infection to solve the resistance and remaining side-effect problems that are the major weak points of traditional antibiotics.

Evaluation on Antimicrobial Activity of Psoraleae semen Extract Controlling the Growth of Gram-Positive Bacteria

  • Kim, Sejeong;Lee, Soomin;Lee, Heeyoung;Ha, Jimyeong;Lee, Jeeyeon;Choi, Yukyung;Oh, Hyemin;Hong, Joonbae;Yoon, Yohan;Choi, Kyoung-Hee
    • Food Science of Animal Resources
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    • v.37 no.4
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    • pp.502-510
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    • 2017
  • This study investigated bacterial growth-inhibitory effect of 69 therapeutic herbal plants extracts on 9 bacterial strains using a disc diffusion assay. Especially, the antimicrobial activity of Psoraleae semen, which showed different activity on pathogenic Gram-positive and Gram-negative bacteria, was evaluated by MIC (minimal inhibition concentration) and biofilm formation assay. The effect of Psoraleae semen extract on bacterial cell membranes was examined by measurement of protein leakage (optical density at 280 nm) and scanning electron microscope (SEM). No clear zone was formed on discs containing Gram-negative bacteria, but Gram-positive bacteria exhibited clear zones. The MICs of Psoraleae semen extract were $8{\mu}g/mL$ for Streptococcus mutans, and $16{\mu}g/mL$ for Enterococci and Staphylococcus aureus. In addition, biofilm formation was inhibited at concentration $8-16{\mu}g/mL$. Protein leakage values and SEM images revealed that cell membranes of Gram-positive bacteria were impaired following exposure to the extract. Further, the extract inhibited the growth of Listeria monocytogenes in sausages. These results indicate that Psoraleae semen extract could be utilized as a natural antimicrobial agent against Gram-positive bacteria.

Effects of the growth inhibition against plant pathogenic bacteria using Beauveria bassiana (백강균(Beauveria bassiana)을 이용한 식물병원성세균 생장억제효능 검증)

  • Lee, Ki Man;Nam, Sung Hee;Hong, In Pyo;Sung, Gyoo Byung;Bae, Yoon Hwan;Kang, Tae Jin
    • Journal of Mushroom
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    • v.9 no.2
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    • pp.87-89
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    • 2011
  • This study was performed to investigate the antibacterial activities and the minimal inhibitory concentration (MIC) of 6 strains of Beauveria bassiana against mulberry pathogenic bacteria. The antibacterial activities and the MIC were measured using paper disc method and broth dilution method, respectively. The antibacterial activities were found out just B. bassiana J200, and shown at 13 mm from Erwinia rhapontici KACC 10407 and at 17 mm from Pseudomonas syringae KACC 10390 and Xanthomonas campestris KACC 12134. The MIC were all observed at 4.0% from E. rhapontici KACC 10407, P. syringae KACC 10390 and X. campestris KACC 12134. The results suggest that B. bassiana could play a good role for biological control against mulberry pathogenic bacteria.

Effects of Environmental Factors on Nitrite Accumulation in a Strong Nitrogen Removal System (고농도 질소폐수 처리 공정에서 환경인자가 아질산염 축적에 미치는 영향)

  • Park, Noh-Back;Choi, Woo-Yung;Yoon, Ae-Hwa;Jun, Hang-Bae;Park, Sang-Min
    • Journal of Korean Society of Water and Wastewater
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    • v.24 no.1
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    • pp.51-62
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    • 2010
  • The high concentration of N in the wastewater from livestock farming generally renders the efficiency of the wastewater treatment. Therefore, removal of N in livestock wastewater is crucial for successful treatment. The current study was conducted to investigate the optimum conditions for partial nitrification under anaerobic condition following nitritation in TPAD-BNR(two-phase anaerobic digestion-biological nitrogen removal) operating system. Sequential operating test to stimulate partial nitrification in reactor showed that partial nitrification occurred at a ratio of 1.24 in $NO_2{^-}$-N:$NH_4{^+}$-N. With this result, a wide range of factors affecting stable nitritation were examined through regression analysis. In the livestock wastewater treatment procedure, the hydraulic retention time (HRT) and pH range for optimum nitrite accumulation in the reactor were 1-1.5 days and 7-8, respectively. It was appeared that accumulation of $NO_2{^-}$-N in the reactor is due to inhibition of the $NO_2{^-}$-N oxidizer by free ammonia (FA) while the effect of free nitrous acid was minimal. Nitrification was not influenced by DO concentration at a range of 2.0-3.0 mg/L and the difference in the growth rate between $NH_4{^+}$-N oxidizer and $NO_2{^-}$-N oxidizer was dependent on the temperature in the reactor.

In Vitro Antibacterial Effects of Wandae-tang Aqueous Extracts and Their Combination Effects with Clindamycin against Gardnerella Vaginalis (완대탕(完帶湯)의 Gardnerella vaginalis에 대한 시험관내 항균력 및 Clindamycin과 병용효과)

  • Lee, Seung-Hye;Kim, Dong-Chul
    • The Journal of Korean Obstetrics and Gynecology
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    • v.25 no.1
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    • pp.34-46
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    • 2012
  • Objectives: The object of this study was to observe the in vitro antibacterial effects of Wandae-tang extracts and combination of Wandae-tang extracts and Clindamycin against Gardnerella vaginalis ATCC14018. Methods: Antibacterial activities against Gardnerella vaginalis ATCC14018 of Wandae-tang extracts were detected using standard agar microdilution methods. In addition, the effects on the bacterial growth curve were also monitored at minimal inhibitory concentration(MIC) and $MIC{\times}2$ levels. The combination effects of Wandae-tang extracts with Clindamycin were observed by Checkerboard microtiter assay, and the effects of bacterial growth curve was treated with Wandae-tang extracts MIC+Clindamycin MIC, 1/2 MIC and 1/4 MIC, respectively. Results: MIC of Wandae-tang extracts and Clindamycin against Gardnerella vaginalis ATCC14018 were detected as $1.719{\pm}0.856$(0.782~3.125) $mg/m{\ell}$ and $0.010{\pm}0.006$ (0.004~0.016) ${\mu}g/m{\ell}$. In addition, Clindamycin and Wandae-tang extracts were also showed marked dosage-dependent inhibition of bacterial growth, and more dramatical inhibitions were detected in Clindamycin+Wandae-tang extracts MIC treatment. Fractional inhibitory concentration index in combination of Wandae-tang extracts and Clindamycin were detected as $0.294{\pm}0.052$(0.250~0.375) at Checkerboard microtiter assay. Conclusions: The results obtained in this study suggest that Wandae-tang extracts showed antibacterial effects against Gardnerella vaginalis ATCC14018, and they also showed dosage-dependent inhibitory effects on the bacterial growth. In addition, combination treatment of Wandae-tang extracts with Clindamycin showed more synergistically potent inhibitory effects on the growth of Gardnerella vaginalis.