• Title/Summary/Keyword: dilution method

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Antibacterial Effect of Haedokgeumhwa-san against Methicillin-Resistant Staphylococcus aureus (해독금화산(解毒金花散)의 Methicillin-Resistant Staphylococcus aureus에 대한 항균효과)

  • Lee, Ha-Il;Lee, Su-Kyung;Kwon, Young-Mi;Song, Yung-Sun
    • Journal of Korean Medicine Rehabilitation
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    • v.25 no.2
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    • pp.1-13
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    • 2015
  • Objectives Methicillin-Resistant Staphylococcus aureus (MRSA) is a human pathogen and a major cause of hospital-acquired infections. New antibacterial agents that have not been compromised by bacterial resistance are needed to treat MRSA-related infections. In this study, we investigated the antimicrobial activity ofethanol extract of Haedokgeumhwa-san (HGH) which prescription is composed of korean medicine against MRSA. Methods The antibacterial activity of HGH extract was evaluated against MRSA strains by using the Disc diffusion method, broth microdilution method (minimal inhibitory concentration; MIC), checkerboard dilution test, and time-kill test; its mechanism of action was investigated by bacteriolysis, detergent or ATPase inhibitors. The checkerboard dilution test was used to examined synergistic effect of ampicillin, oxacillin, ciprofloxacin, vancomycin, gentamicin and norfloxacin in combination with HGH ethanol extract. A time-kill assay was performed a survival curve which was obtained by plotting viable colony counts depending on time on bacterial growth. Results The minimum inhibitory concentration (MIC) of ethanol extract (HGH) ranged from 1,000 to $2,000{\mu}g/mL$ against all the tested bacterial strains, respectively. We are able to confirm that HGH extract has potentially strong antibacterial activity. In the checkerboard dilution test, fractional inhibitory concentration index of HGH in combination with antibiotics indicated synergy or partial synergism against S. aureus. A time-kill study showed that the growth of the tested bacteria was considerably inhibited after 8 hr of treatment with the combination of HGH with selected antibiotics. For measurement of cell membrane permeability, HGH $250{\sim}1,000{\mu}g/mL$ along with concentration of Triton X-100 (TX) and Tris-(hydroxymethyl) aminomethane (Tris) were used. In the other hand, N,N-dicyclohexylcarbodimide (DCCD) and Sodium azide ($NaN_3$) was used as an inhibitor of ATPase. TX, Tris, DCCD and $NaN_3$ cooperation against S. aureus showed synergistic action. Accordingly, antimicrobial activity of HGH was affected by cell membrane and inhibitor of ATPase. Conclusions These results suggest that Haedokgeumhwa-san extract has antibacterial activity, and that HGH extract offers a potential as a natural antibiotic against MRSA.

Degradation of Polyvinyl Alcohol in Dye-Processing Wastewater by Agar-Acrylamide Microbial Immobilization Method (한천-아크릴아마이드 미생물 고정화법에 의한 폐수 중 폴리비닐알콜의 분해)

  • 김재훈;김정목조무환
    • KSBB Journal
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    • v.10 no.3
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    • pp.241-248
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    • 1995
  • For the treatment of poorly biodegradable polyvinyl alcohol(PVA) in dye-processing wastewater, immobilized microbial beads were prepared by uslng agar-acrylamide method. PVA removal efficiency for the synthetic wastewater was 85% at the PVA volume loading rate of $3.1g/\ell$.day. In case of real desizing wastewater, PVA removal efficiency was 81.3% at the PVA volume loading rate of $3.25g/\ell$.day. In observation of cross section of immobilized bead passed 5 months with diameter of 2.4mm, the growth of cell was limited by the resistance of substrate and oxygen transfer for the inners region of more than 48% of bead radius from the surface. It was estimated that 70% of total removed PVA was degraded by the immobilized cells in the continuous immobilized reactor. Substrate utilization rate in the suspended reactor was decreased with increasing dilution rates above 0.083 hr-1, but that in the immobilized reactor was increased with increasing dilution rates up to 0.125hr-1. The substrate removal efficiency of immobilized reactor was much superior to that of suspended reactor with increasing dilution rates. Saturation constant of substrate utilization rate equation, Ks was $6.6 g PVA/\ell$, and maximum specific substrate utilization. k was 0.175g PVA/g cell.hr

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Antibacterial activity of Tonghyeonipal-dan against Methicillin-resistant Staphylococcus aureus (통현이팔단 에탄올 추출물의 Methicillin Resistant Staphylococcus aureus에 대한 항균활성)

  • KIM, In-Won;KANG, Ok-Hwa;KONG, Ryong;KWON, Dong-Yeul
    • The Korea Journal of Herbology
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    • v.30 no.5
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    • pp.15-21
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    • 2015
  • Objectives : Methicillin-resistantStaphylococcus aureus(MRSA) is a human pathogen. New antibacterial agents are needed to treat MRSA-related infections. This study investigated the antibacterial activity of EtOH 70% extracts ofTonghyeonipal-dan(THD) which prescription is composed of oriental medicine against MRSA.Methods : The antibacterial activity of THD was evaluated against MRSA strains by using the Disc diffusion method, broth microdilution method, Checkerboard dilution test, and Time-kill test; its mechanism of action was investigated by bacteriolysis, detergent or ATPase inhibitors were used.Results : The minimum inhibitory concentration (MIC) of THD is 1,000~2,000 μg/mL against MRSA. In the checkerboard dilution test, fractional inhibitory concentration index (FICI) of THD in combination with antibiotics indicated synergy or partial synergism againstS. aureus. Furthermore, a time-kill assay showed that the growth of the tasted bacteria was considerably inhibited after 24 h of treatment with the combination of THD with selected antibiotics. For measurement of cell membrane permeability, THD 500 μg/mL along with concentration of Triton X-100 (TX) and Tris-(hydroxymethyl) aminomethane (TRIS) were used. In the other hand, N,N-dicyclohexylcarbodimide (DCCD) and Sodium azide (NaN3) were used as an inhibitor of ATPase. TX, TRIS, DCCD and NaN3 cooperation againstS. aureusshowed synergistic action.Conclusions : Accordingly, antimicrobial activity of THD was affected by cell membrane and inhibitor of ATPase were assessed. These results suggest that THD has antibacterial activity, and that THD extract offers great potential as a natural antibiotic against MRSA.

Determination of Lead in Steels by Hydride generation-Inductively Coupled Plama/Mass Spectrometry (수소화물 발생-유도결합플라스마 질량분석법에 의한 철강 중의 납의 정량)

  • Park, Chang Joon;Song, Sun Jin;Lee, Dong Soo
    • Analytical Science and Technology
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    • v.14 no.5
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    • pp.410-415
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    • 2001
  • An analytical method has been developed which determines lead in steel samples by inductively coupled plasma mass spectrometry (ICP-MS) with sample introduction by the hydride generation. The lead hydride is not stable and requires and oxidant for the oxidation into metastable Pb(IV) before reduction to $PbH_4$ with $NaBH_4$. A study was carried out to find and optimum lead hydride generation condition for a sample solution with more than $1000{\mu}g/mL$ Fe matrix. $K_2Cr_2O_7$ was found to work as an efficient oxidant when more than $10{\mu}g/mL$ Fe matrix was present. Lactic acid was used with the oxidant as a complexing agent of the metastable Pb(IV) to enhance sensitivity. Optimum concentrations of the sample acidity, oxidant and lactic acid were different depending on the matrix concentration. The isotope dilution method was employed for the quantitation of lead. The determined Pb concentrations of the NIST steel SRM 361 and 362 were in good agreement with the certified values within the uncertainty range.

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Effects of Plantain (Plantago lanceolata L.) Herb and Heat Exposure on Plasma Glucose Metabolism in Sheep

  • Al-Mamun, M.;Tanaka, C.;Hanai, Y.;Tamura, Y.;Sano, H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.6
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    • pp.894-899
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    • 2007
  • An experiment was conducted using a [6, 6-$^2H$]glucose isotope dilution method to determine the effects of plantain (Plantago lanceolata L.) on plasma glucose metabolism in sheep taken from a thermoneutral environment and exposed to a hot environment. The sheep were fed either mixed hay (MH) of orchardgrass (Dactylis glomerata L.) and reed canarygrass (Phalaris arundinacea L.) at a 60:40 ratio or MH and plantain (PL) at a 9:1 ratio in a crossover design for each 23-day period. In both dietary treatments the metabolizable energy (ME) and crude protein intake were designed to be isoenergetic and isoproteinous at around maintenance level. The sheep were taken from a thermoneutral environment ($20^{\circ}C$, 70% RH) and exposed to a hot environment ($28-30^{\circ}C$, 70% RH) for 5 days. The isotope dilution method using a single injection of [6, 6-$^2H$]glucose was performed on the $18^{th}$ day of the thermoneutral environment and on the $5^{th}$ day of heat exposure. Plasma glucose pool size was numerically lower (p = 0.26) during heat exposure on both dietary treatments, and numerically higher (p = 0.13) on the MH diet irrespective of environmental temperature. Plasma NEFA concentration (p = 0.01) and glucose turnover rate (p = 0.03) were decreased during heat exposure, but remained similar between diets. It could be concluded that, although no positive impact of plantain on glucose metabolism was found under the present experimental conditions (plantain constituted only 10% of basal diet), plantain herb is an alternative to MH for rearing sheep in both thermoneutral and hot environments.

Comparative Study Using MIC Test for Cefazedone and other First Generation Cephalosporins (1세대 세파계 항생제인 Cefazedone의 MIC Test를 통한 비교 연구)

  • Bok, Hae-Sook;Kim, Myung-Min;Choi, Kyung-Eob
    • Korean Journal of Clinical Pharmacy
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    • v.5 no.2
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    • pp.13-16
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    • 1995
  • The Minimum Inhibitory Concentration (MIC) of a first-generation cephalosporin derivative, Cefazedone (CZD; $PAZERON^R$ inj.) was determined by the two-fold serial agar dilution method. The in-vitro antibacterial activity of CZD against a wide variety of clinical isolates was compared with those of other first generation cephalosporins such as Methylol Cephalexin (CEX), Cefazolin (CEZ), Cefadroxil (CDX), Cephradine (CED), Ceftezol (CTZ) and one of second generation cephalsporin antibiotics, Cefotaxime (CTX). CZD had the most potent inhibitory effect against Gram-positive strains, when compared to the first-generation cephalosporin antibiotics tested in this study and CTX. The geometric MIC mean of CZD for Gram-positive strains was calculated as 0.386 kg/m{\ell}$, and those of CEX, CEZ, CDX, CTZ, CED, and CTX were 6.073, 0.894, 3.399, 0.748, 7.884 and 1.502 $kg/m{\ell}$, respectively. In addition, the geometric mean of CZD for staphylococclJs aureus strains was obtained as 0.340 $kg/m{\ell}$ and those of CEX, CEZ, CDX, CTZ, CED, and CTX 6.145, 0.534, 4.126, 0.442, 10.51, and 2.500 $kg/m{\ell}$, respectively. Against Gram-negative strains, CZD showed better antibacterial activity than CEZ, CDX, CTZ, and CED.

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Effects of Rice Straw Supplemented with Urea and Molasses on Intermediary Metabolism of Plasma Glucose and Leucine in Sheep

  • Alam, Mohammad Khairul;Ogata, Yasumichi;Sato, Yukari;Sano, Hiroaki
    • Asian-Australasian Journal of Animal Sciences
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    • v.29 no.4
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    • pp.523-529
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    • 2016
  • An isotope dilution method using $[U-^{13}C]glucose$ and $[1-^{13}C]leucine$ (Leu) was conducted to evaluate the effects of rice straw supplemented with urea and molasses (RSUM-diet) on plasma glucose and Leu turnover rates in sheep. Nitrogen (N) balance, rumen fermentation characteristics and blood metabolite concentrations were also determined. Four sheep were fed either mixed hay (MH-diet), or a RSUM-diet with a crossover design for two 21 days period. Feed allowance was computed on the basis of metabolizable energy at maintenance level. The isotope dilution method was performed as the primed-continuous infusion on day 21 of each dietary period. Nitrogen intake was lower (p = 0.01) for the RSUM-diet and N digestibility did not differ (p = 0.57) between diets. Concentrations of rumen total volatile fatty acids tended to be higher (p = 0.09) for the RSUM-diet than the MH-diet. Acetate concentration in the rumen did not differ (p = 0.38) between diets, whereas propionate concentration was higher (p = 0.01) for the RSUM-diet compared to the MH-diet. Turnover rates as well as concentrations of plasma glucose and Leu did not differ between diets. It can be concluded that kinetics of plasma glucose and Leu metabolism were comparable between the RSUM-diet and the MH-diet, and rumen fermentation characteristics were improved in sheep fed the RSUM-diet compared to the MH-diet.

Geometric Sensitivity Index for the GNSS Using Inner Products of Line of Sight Vectors

  • Won, Dae Hee;Ahn, Jongsun;Sung, Sangkyung;Lee, Chulsoo;Bu, Sungchun;Jang, Jeagyu;Lee, Young Jae
    • International Journal of Aeronautical and Space Sciences
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    • v.16 no.3
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    • pp.437-444
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    • 2015
  • Satellite selection and exclusion techniques have been applied to the global navigation satellite system (GNSS) with the aim of achieving a balance between navigational performance and computational efficiency. Conventional approaches to satellite selection based on the best dilution of precision (DOP) are excessively computational and complicated. This paper proposes a new method that applies a geometric sensitivity index of individual GNSS satellites. The sensitivity index is derived using the inner product of the line of sight (LOS) vector of each satellite. First, the LOS vector is computed, which accounts for the geometry between the satellite and user positions. Second, the inner product of each pair of LOS vectors is calculated, which indicates the proximities of the satellites to one another. The proximity can be determined according to the sensitivity of each satellite. A post-processing test was conducted to verify the reliability of the proposed method. The proposed index and the results of a conventional approach that measures the dilution of precision (DOP) were compared. The test results demonstrate that the proposed index produces results that are within 96% of those of the conventional approach and reduces the computational burden. This index can be utilized to estimate the sensitivity of individual satellites, obtaining a navigation solution. Therefore, the proposed index applies to satellite selection and exclusion as well as to the sensitivity analyses of multiple GNSS applications.

Antibacterial Activity and Inhibition of Resistance in Methicillin-resistant Staphylococcus aureus by Maneung-hwan Ethanol Extract (만응환(萬應丸) 에탄올 추출물의 메티실린 내성 포도상구균에 대한 항균활성 및 내성억제 효과)

  • Na, Yong-su;Kim, Jong-gyu;Song, Yung-sun
    • Journal of Korean Medicine Rehabilitation
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    • v.30 no.1
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    • pp.31-45
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    • 2020
  • Objectives In this study, we investigated the antimicrobial activity of a 70% ethanol extract of Maneung-hwan (MEH), which is prescribed by practitioners of oriental medicine for use against methicillin-resistant Staphylococcus aureus (MRSA). Methods The antibacterial activity of MEH against MRSA strains was evaluated using the disc diffusion method, broth microdilution method (minimal inhibitory concentration, MIC), checkerboard dilution test, and time-kill test. The mechanism of action of MEH was investigated by bacteriolysis using detergents or ATPase inhibitors Additionally, mRNA and protein expression were investigated by quantitative reverse transcription-polymerase chain reaction and western blot assay, respectively. Results The MIC of MEH was 25~1,600 ㎍/mL against all the tested bacterial strains. We showed that MEH extract exerts strong antibacterial activity. In the checkerboard dilution test, the fractional inhibitory concentration index of MEH in combination with antibiotics indicated synergism or partial synergism against S. aureus. The time-kill study indicated that the growth of the tested bacteria was considerably inhibited after a 24-h treatment with MEH and selected antibiotics. To measure the cell membrane permeability, MEH (3.9 ㎍/mL) was combined with Triton X-100 (TX) at various concentrations N,N-dicyclohexylcarbodimide (DCCD) was also tested as an ATPase inhibitor. TX and DCCD cooperation against S. aureus exhibited synergistic action. Accordingly, the antimicrobial activity of MEH in the context of cell membrane rupture and ATPase inhibition was assessed. Additionally, the expression of genes and proteins associated with resistance was reduced after exposing MRSA to MEH. Conclusions These results suggest that MEH possesses antibacterial activity and acts as a potential natural antibiotic against MRSA.

Drug susceptibility of bacteria and M pachydermatis isolated from canine external ear canals (개 외이도에서 분리한 세균 및 M pachydermatis의 약제감수성에 대하여)

  • Kim, Ki-hyang;Choi, Won-phil;Yeo, Sang-geon
    • Korean Journal of Veterinary Research
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    • v.39 no.3
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    • pp.560-565
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    • 1999
  • The present work was conducted to investigate the drug susceptibility of microorganisms isolated from canine external ear canals. Antifungal susceptibility test of M pachydermatis (17 strains) was perfomed by agar dilution method, using 11 antifungal drugs including amphotericin B(A), nystatin(N), pimaricin(P), griseofulvin(G), bifonazole(B), clotrimazole(C), miconazole(M), econazole(E), ketoconazole(K), tolnaftate(T), 5-fluorocytosine(F). All isolates were highly sensitive to K, M, T(geometric mean MIC ; GM $MIC{\leq}0.16{\mu}g/ml$) but they weren't sensitive to P, F and G(GM $MIC{\geq}92.37{\mu}g/ml{\sim}{\geq}128{\mu}g/ml$). Antibacterial susceptibility test against 119 isolates of bacteria was performed by agar dilution method, using 9 antibacterial drugs including erythromycin(ET), chloramphenicol(CP), gentamycin(G), vancomycin(V), ampicillin(AP), amoxacillin(AX), chlortetracycline(CT), ciprofloxacin(CF), enrofloxacin(EF). All isolates of Staphylococcus spp(101 strains) were highly sensitive to EF, CF, G(GM MIC $0.33{\sim}1.47{\mu}g/ml$). In other gram positive cocci(4 strains), they were highly sensitive to EF, CF, V(GM MIC $1{\sim}4.76{\mu}g/ml$) and CT(GM MIC 1 UFL unit/ml). In gram positive rods(13 strains), they were highly sensitive to EF, CF, G(GM $MIC{\leq}0.19{\sim}1{\mu}g/ml$). In Pseudomonas aeruginosa(1 strain), it was highly sensitive to AX, EF, ET, CF(GM MIC $0.06{\sim}1{\mu}g/ml$) and CT(GM MIC 1 UFL unit/ml). All isolates weren't sensitive to AP(GM MIC 16~>$32{\mu}g/ml$).

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