• Title/Summary/Keyword: EDTA inhibition

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Evaluation of EDTA-based Three Methods to Detect IMP-1 and VIM-2 Type Metallo-${\beta}$-Lactamase-Producing Clinical Isolates of Imipenem Resistant Acinetobacter and Pseudomonas spp.

  • Hong, Seung-Bok;Shin, Kyung-A;Hwang, Seock-Yeon
    • Biomedical Science Letters
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    • v.17 no.2
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    • pp.135-140
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    • 2011
  • We compared three EDTA-based phenotypic screening methods for detecting IMP-1 and VIM-2 type metallo-${\beta}$- lactamase (MBL)-producing isolates of Acinetobacter and Pseudomonas spp., EDTA-double disk synergy test (EDTADDST), Etest MBL, and imipenem (IPM)-EDTA disk test. A total of 183 isolates (65 Acinetobacter spp. and 118 Pseudomonas spp. showing IPM resistance), confirmed to MBL genes by PCR, were used. The criteria for MBL production were (i) presence of a synergistic zone between IPM and EDTA disks in EDTA-DDST, (ii) reduction of IPM minimal inhibitory concentration by ${\geq}$ 3 twofold dilutions in the presence of EDTA in the Etest MBL, and (iii) ${\geq}$ 7 mm increase in the inhibition zone around the IPM plus EDTA disks compared with a sole IPM disk in the IPM-EDTA disk test. In this study using 87 MBL-producing and 96 MBL-nonproducing isolates, the sensitivities/specificities of EDTA-DDST, Etest MBL and IPM-EDTA disk tests were 94.3/78.1%, 89.7/91.7%, and 97.7/95.8%, respectively. When the threshold for the increase of the inhibition zone around the IPM plus EDTA disk over a sole IPM disk was altered to ${\geq}$ 5 mm and ${\geq}$ 8 mm for Acinetobacter spp. and Pseudomonas spp., respectively, the sensitivity and specificity of the test were 98.9% and 96.9%, respectively. Of the three EDTA-based phenotypic tests, the IMP-EDTA disk test was superior for detection of MBL-producing isolates.

Potential Role of $Ca^{++}$ on the Differentiation of Erythroid Progenitor Cells

  • Cho, In-Koo;Huh, In-Hoe;Lee, Sang-Jun;Kim, Dong-Seop;Ann, Hyung-Soo
    • Archives of Pharmacal Research
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    • v.18 no.2
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    • pp.105-112
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    • 1995
  • In ordedr to gain insight into the mechanisms byl which erythropoietin promotes erythropoiesis, effects of various inhibitors on the erythropoietin-propmoted differentiation of erythroid progenitor cells and on the erythroid progenitor cells and on the erythropoietin-promoted $Ca^{++}$ uptake in the progenitor cells were determined, and the relationship between the inhibitory activity of each inhibitor cells were determined, and he relationship between the inhibitory activity of each inhibitor toward the differentiation and channel blocker (varapamil), a $Ca^{++}$ chelator (EDTA) and a protein kinase C inhibitor (stauroporine). All of these agents inhibited both the erythropoietin-mediated differentiation of the erythroid progenitor cells, as determined by the incroporation of $^{59}Fe$ into heme, and $Ca^{++}$ uptake in a concentrtion dependent manner. In the cases of varapamil and EDTA, the half-miximal inhibitory concentration $(IC_{50})$ values for differentiation of the progenitor cells may be theconsequence of the inhibition of the $Ca^{++}$ uptake in a concentration dependent manner. In the cases of varapamil and EDTA, the half-miximal inhibitory concentration dependent manner. In the cases of verapamil and EDTA, the half-miximal inhibitory concentration $(IC_{50})$ values for differentiation of the progenitor cells may be the consequence of the inhibition of the $Ca^{++}$ uptake by the inhibitor. On the other hand, in the cases of genistein and stauroporine, the $IC_{50}$ values for inhibition of differentitation were significantly different from that for inhibition of $Ca^{++}$uptake. These results suggest that the mechanism of inhibition of differentiation by these two inhibitors in complex. However, taken all together, the above results support the proposition that $Ca^{++}$ uptake may play a role in the erythropoietin-mediated differentiation of erythoid progenitor cells.

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TiO2 Reuse and Recovery from the Photocatalytic Oxidation of Cu(II)-EDTA using TiO2/UV-A System (TiO2/UV-A 시스템을 이용한 Cu(II)-EDTA의 광촉매 산화반응에서 TiO2 재사용 및 회수)

  • Lee, Seung-Mok
    • Journal of Korean Society on Water Environment
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    • v.21 no.1
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    • pp.84-91
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    • 2005
  • $TiO_2-catalyst$ suspensions work efficiently in Photocatalytic oxidation (PCO) for wastewater treatment. Nevertheless, once photocatalysis is completed, separation of the catalyst from solution becomes the main problem. The PCO of Cu(II)-EDTA was studied to determine the reusability of the titanium dioxide catalyst. Aqueous solutions of $10^{-4}M$ Cu(II)-EDTA were treated using illuminated $TiO_2$ particles at pH 6 in a circulating reactor. $TiO_2$ was reused in PCO system for treatment of Cu(II)-EDTA comparing two procedures: reuse of water and $TiO_2$ and reuse of the entire suspension after PCO of Cu(II)-EDTA. The results are as follows; (i) Photocatalytic efficiency worsens with successive runs when catalyst and water are reused without separation and filtration, whereas, when $TiO_2$ is separated from water, the reused $TiO_2$ is not deactivated. (ii) The $TiO_2$ mean recovery (%) with reused $TiO_2$ was 86.4%(1.73g/L). Although the mean initial degradation rate of Cu(II)-EDTA and Cu(II) was lower than that using fresh $TiO_2$, there was no significant change in the rate during the course of the three-trial experiment. It is suggested that Cu(II)-EDTA could be effectively treated using an recycling procedure of PCO and catalyst recovery. (iii) However, without $TiO_2$ separation, the loss of efficiency of the PCO in the use of water and $TiO_2$ due to Cu(II), DOC remained from previous degradation and Cu(II)-EDTA added to the same suspension was observed after 2 trials, and resulted in the inhibition of the Cu(II)-EDTA, Cu(II) and DOC destruction.

Enhanced Phytoremediation by Echinochloa crus-galli in Arsenic Contaminated Soil in the Vicinity of the Abandoned Mine (폐광지역 비소오염 토양에 대한 피(Echinochloa crus-galli)를 이용한 보강된 식물상복원공법)

  • Park, Ji-Yeon;Kim, Ju-Yong;Lee, Byung-Tae;Kim, Kyoung-Woong;Lee, Jin-Soo
    • Economic and Environmental Geology
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    • v.43 no.2
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    • pp.101-107
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    • 2010
  • In order to deal with the problem that phytoremediation takes long time in achieving the practical effect, the enhanced phytoremediation by Barnyard grass (Echinochloa crus-galli) was conducted. In addition, we examined the synergistic effect by adding PSM (phosphate -solubilizing microbes) and EDTA (ethylenediaminetetraacetic acid) to the arsenic contaminated soil in the vicinity of the abandoned mine. The removal efficiency of arsenic in the site with PSM application increased about 16% when compared to control site, which was due to increase of plant biomass. The EDTA has been successfully utilized in respect of enhanced mobility and solubility of arsenic in the soil. As a result, BF (bioaccumulation factor) significantly increased but the inhibition of plant growth resulted in 20% reduction of arsenic removal efficiency. The application of PSM and EDTA may enhance the efficiency of phytoremediation. However, the time and method of EDTA application should be further examined to reach the maximum removal efficiency.

Inhibition of Human Neutrophil Elastase by NSAIDs and Inhibitors, and Molecular Pharmacological Mechanism of the Inhibition (비스테로이드성 항염증제와 효소 억제제에 의한 사람 중성구 Elastase의 활성도 억제 및 분자약리학적 기전)

  • Kang, Koo-Il;Kim, Woo-Mi;Hong, In-Sik;Lee, Moo-Sang
    • The Korean Journal of Pharmacology
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    • v.32 no.3
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    • pp.425-431
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    • 1996
  • Human neutrophil elastases (HNElastase, EC 3.4.21.37), a causative factor of inflammatory diseases, are regulated by plasma proteinase inhibitors, alpha-proteinase inhibitor and ${\alpha}_2-macroglobulin$. Under certain pathological conditions, however, released enzymes or abnormal function of inhibitors may cause various inflammatory disease. NSAIDs have been clinically applied for treatment of inflammatory diseases. Inhibition of cyclooxygenase is a known mechanism of action of NSAIDs in the treatment of inflammatory disease. In in vitro experiments, HNElastase was inhibited by naproxen, phenylbutazone, and oxyphenbutazone, but ibuprofen, ketoprofen, aspirin, salicylic acid, and tolmetin did not inhibit elastase. HNElastase was also inhibited by chelating agents, EDTA & EGTA, and tetracyclines. Removal of divalent metal ions by EDTA caused inhibition of elastase, and reconstitution of the metal ions recovered the enzyme activity to a certain level. Frequencies and contours in the Raman spectra of various conditions of human neutrophil elastase undergo drastic changes upon partial removal and/or reconstitution of calcium and zinc ions. The metal ion content dependent activities and change of the contour of the Raman spectrogram suggest us that the mechanism of action of a chelator or chelator-like agents on neutrophil elastase may be related to the conformational change at/or near the active site, especially -C=O radical or -COOH radical.

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Effect of Lysozyme, Glycine, Lysine and EDTA on the Growth of Lactobacillus plantarum (Lysozyme과 Glycine, Lysine 및 EDTA의 첨가가 Lactobacillus plantarum의 성장에 미치는 영향)

  • Lee, Sung-Ki;Kim, In-Ho;Yoo, Ick-Jong
    • Korean Journal of Food Science and Technology
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    • v.24 no.1
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    • pp.11-13
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    • 1992
  • Effect of 200 ppm lysozyme. 0.1 glycine and 0.1% lysine on inhibition growth of Lactobacillus plantarum was investigated in MRS broth. All samples except control were effective in inhibiting the growth and especially the combination of lysozyme and glycine was observed to be highly effective. The mixture effect against microbial growth was increased as concentration of lysozyme with glycine or lysozyme with EDTA was respectively increased. Lactobacillus plantarum almost didn't grow in MRS broth containing lysozyme(>200 ppm) with glycine(>0.5%), or lysozyme(>200 ppm) with EDTA(> 0.8 mM). It was found that the growth of L. plantarum could be extremely inhibited in 120 ppm lysozyme with at least 0.8 mM EDTA compared with control.

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Studies on the Myofibrilar Protein from Korean Duck Muscle (오리고기의 근원섬유 단백질에 관한 연구)

  • Chang, In-Yae;Nam, Hyun-Keun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.9 no.1
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    • pp.45-50
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    • 1980
  • Myofibrilar Protein from Korean Duck Muscle was extracted and ATPase activities were studied. The results were as follows: 1. Mg-activated ATPase activity of Myofibril from Korean Duck, muscle exhibited a biphasic response, ATPase activity was high at a low ionic strength and low activity was showed at high ionic strength. 2. Effect of EDTA on the Myofibrillar protein ATPase activity was studied, it was investigated that the EDTA inhibition was showed at the concentration of $6.9{\mu}g$ and it above. 3. It showed that the effect of Ca++ on ATPase activity was decreased at the lower than $3{\mu}g$. Inhibition showed at the concentration of $6.9{\mu}g$ and it above. 4. It showed that the effect of Mg++ on ATPase activity was decreased at the lower than $3.6{\mu}g$. Inhibition showed at the concentration of $3.9{\mu}g$ and it above.

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[$Zn^{2+}$ Modulates the Responses of Rat Dorsal Horn Neuron to C-Fiber Stimulation and Excitatory Amino Acids

  • Ahn, Chang-Hoon;Shin, Hong-Kee;Kim, Jin-Hyuk
    • The Korean Journal of Physiology and Pharmacology
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    • v.4 no.6
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    • pp.455-461
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    • 2000
  • Zinc contained in the neurons of central nervous system is activity-dependently released and then attenuates NMDA (N-methyl-D-aspartate)-induced neurotoxicity while augmenting non-NMDA-induced neurodegeneration. Zinc also has been reported to produce antinociceptive action on the inflammation- and nerve injury-induced hyperalgesia in the behavioral test. In this study, we investigated the effects of zinc on the responses of dorsal horn cells to NMDA, kainate and graded electrical stimulation of C-fibers. In the majority of WDR cells (70.6%), zinc current-dependently inhibited WDR cell responses to NMDA and in the remaining cells, produced biphasic responses; excitation followed by inhibition. Zinc augmented the responses of WDR cells to iontophoretical application of kainate. The dominant effect of $Zn^{2+}$ on the responses of WDR cells to C-fiber stimulation was excitatory, but inhibition, excitation-inhibition and no change of the responses to C-fiber stimulation were induced. $Ca^{2+}-EDTA$ antagonized the excitatory or inhibitory effects of $Zn^{2+}$ on the WDR cell responses. These experimental findings suggest that $Zn^{2+}$ modulates the transmission of sensory information in the rat spinal cord.

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Inhibition of Enzymatic Degradation of Leucine Enkephalin and $[D-Ala^2]$-Leucine Enkephalinamide in Various Rabbit Mucosal Extracts by Inhibitors (효소 억제제에 의한 토끼의 점막 추출액중 로이신엔케팔린 및 [D-알라$^2$-로이신엔케팔린아미드의 분해 억제)

  • Chun, In-Koo;Park, In-Sook;Hyun, Jeen
    • Journal of Pharmaceutical Investigation
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    • v.26 no.3
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    • pp.175-185
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    • 1996
  • To inhibit the enzymatic degradation of leucine enkephalin (Leu-Enk) and its synthetic analog. $[D-ala^2]$-leucine enkephalinamide (YAGFL), in the nasal, rectal and vaginal mucosal and serosal extracts of rabbits, effects of enzyme inhibitors such as amastatin (AM), puromycin (PM), thiorphan (TP), thimerosal (TM), EDTA, N-carboxymethyl-Phe-Leu (CPL), phenylethyl alcohol (PEA), phenylmercuric acetate (PMA), benzalkonium chloride (BC) and modified cyclodextrins, alone or in combination, were observed by assaying the pentapeptides staying intact during incubation. Mucosa extracts were prepared by exposing freshly-excised mucosal specimens mounted on Valia-Chien cells to isotonic phosphate buffer while stirring. The degradation of Leu-Enk and YAGFL followed the apparent first-order kinetics. The half-lives (mean) in the nasal, rectal and vaginal mucosal extracts were found to be 1.07, 0.33 and 1.14 hr for Leu-Enk, and 16.9, 6.2 and 6.8 hr for YAGFL, respectively. AM or PM, which is an aminopeptidase inhibitor, did not show a sufficient inhibition of Leu-Enk $(50\;{\mu}g/ml)$ degradation in all kinds of extracts. $Dimethyl-{\beta}-cyclodextrin\;(DM-{\beta}-CyD)$ decreased the degradation rate constants of Leu-Enk about 2 or 3 times, comparing with no additive. However, the use of mixed inhibitors of AM $(50\;{\mu}M)$/TM (0.25 mM)/EDTA (5 mM) resulted in a full stabilization of Leu-Enk by decreasing the degradation rate constants 67.3, 161.3 and 113.8 times far the nasal, rectal and vaginal mucosal extracts, respectively, comparing with no inhibitor. With mixed inhibitors, Leu-Enk remained intact more than 90% after 6 hr-incubation. In the stabilization of YAGFL, hM, TP or CPL alone showed little efffct, and some additives demonstrated a considerable inhibition of YAGFL degradation in the rank order of TM > BC > EDTA. However, the addition of mixed inhibitors such as TM (0.5 mM) and EDTA (5 mM) into the extracts protected YAGFL from the degradation by more than 85% even after 24 hr-incubation, suggesting almost complete inhibition of YAGFL degradation in the extract. On the other hand, $DM-{\beta}-CyD\;or\;hydroxypropyl-{\beta}-cyclodextrin$ (10%) were also found to retard enzymatic degradation rates of YAGFL markedly, and resulted in staying intact more than 80% of YAGFL in the nasal and vaginal mucosal extracts, and more than 60% in the rectal mucosal extract after 16 hr-incubation.

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Production of staphylokinase in Staphylococcus hyicus subsp. hyicus strains of swine, poultry and bovine origin (돼지, 닭 및 소유래 Staphylococcus hyicus subsp. hyicus의 staphylokinase 산생능)

  • Park, Joon-seo;Park, Cheong-kyu
    • Korean Journal of Veterinary Research
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    • v.37 no.2
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    • pp.359-365
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    • 1997
  • Staphylococcus hyicus subsp. hyicus strains isolated from pigs, chickens and cattle were examined for the production of staphylokinase after inhibition of staphylococcal proteases by two procedures with EDTA(disodium). In one, EDTA was added to the bovine fibrin-dog plasminogen agar medium in concentration of 0.07% and paper strips soaked in 2mg/ml soy bean trypsin inhibitor were then applied on the agar plates. In the other, paper strips soaked in 5% EDTA solution were applied on the bovine fibrin-dog plasminogen agar plates and the strains to be tested were then streaked at right angles with the strip. By these procedures, staphylokinase activity was detected in 8(88.9%) of 9 strains from diseased pigs and in 57(80.3%) of 71 strains from skin of healthy pigs, but not in any strains from skin of healthy chickens and milk samples of mastitic cattle. Additionally kinase activity in 9 Staphylococcus species and subspecies isolated from bovine intramammary infections was also tested by these procedures. Staphylokinase activity was detected in 74.2% of Staph aureus strains and in 25% of Staph xylosus strains.

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