• Title/Summary/Keyword: sodium azide

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The Role of Abp140p in Actin Dynamics of Budding Yeast

  • Lim, Bum-Soon;Lee, Yong-Keun;Pon, Liza A.;Yang, Hyeong-Cheol
    • International Journal of Oral Biology
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    • v.30 no.1
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    • pp.17-22
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    • 2005
  • In the previous studies of Saccharomyces cerevisiae, Abp140p (actin binding protein 140) fused to GFP has been only a protein that can label actin cables of yeast cells so far. However, the role of Abp140p in actin dynamics was remained elusive. In this study, the function of Abp140p was investigated with a deletion mutant and overexpression of GFP fused Abp140p. The deletion mutant was slightly more susceptible to Latrunculin-A (Lat-A), an actin-monomer sequestering agent, than wild type, although no significant deformation of actin structures was caused by ABP 140 deletion. Overexpression of Abp140p-GFP retarded cell growth, and produced thick and robust actin cables. Lat-A was not able to destabilize the thick actin cables, which suggests that actin dynamics was compromised in the cells with surplus of Abp140p. Therefore, Abp140p seems to stabilize actin cables together with other bundling proteins. Recently, actin cable dynamics of budding yeast was found to have a resemblance to that of filopodial tip of cultured mammalian cells. Retrograde movement of actin cables from buds to mother cells indicated local generation of the cable at bud sites. By using Abp140p-GFP, we traced the steps in the generation of a new actin cable after elimination of old cables by sodium azide. Before the appearance of a new actin cable, Abp140p-GFP concentrated in buds and disappeared, as mother cells became abundant in actin cables. Our observations provide a direct evidence of actin cable formation at buds of budding cells.

Synthesis, Crystal structure, and Magnetic Properties of Dinuclear Iron(III) Complexes with Methoxo Bridges

  • Shin, Jong-Won;Han, Jeong-Hyeong;Rowthu, Sankara Rao;Kim, Bong-Gon;Min, Kil-Sik
    • Bulletin of the Korean Chemical Society
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    • v.31 no.12
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    • pp.3617-3622
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    • 2010
  • The reaction of stoichiometric amount of $FeCl_2{\cdot}4H_2O$, (2-pyridylmethyl, 3-pyridylmethyl)amine (2,3-pyma) and sodium azide/sodium thiocyanate in methanol under aerobic conditions affords the dinuclear Fe(III) complexes, [(2,3-pyma) $(N_3)_2Fe({\mu}-OCH_3)_2Fe(N_3)_2$(2,3-pyma)]${\cdot}CH_3OH$ (1) and [(2,3-pyma)$(NCS)_2Fe({\mu}-OCH_3)_2Fe(NCS)_2$(2,3-pyma)] (2) in good yield. Two bis-methoxy-bridged diiron(III) complexes are isolated and characterized. The coordination geometries around iron(III) ions in 1 and 2 are the same tetragonally distorted octahedron. The iron(III) ions are coordinated by two nitrogens of a 2,3-pyma, two nitrogens of two azide/thiocyanate ions, and two oxygens of two methoxy groups. Both compounds are isomorphous. The structures of 1 and 2 display the C-$H{\cdots}\pi$ and/or $\pi-\pi$ stacking interactions as well as hydrogen bonding interactions, respectively. Compounds 1 and 2 show significant antiferromagnetic couplings through the bridged methoxy groups between the iron(III) ions in the temperature range from 5 to 300 K ($H=-2JS_1{\cdot}S_2$, J=-19.1 and $-13.9\;cm^{-1}$ for 1 and 2).

Types of Extended-Spectrum β-Lactamase Produced in Enteric Bacteria Isolated from Sewage Plant Drain Water (하수처리수에서 분리된 장내세균의 광범위 베타락탐분해효소의 유형)

  • Kim, Gun-Do;Lee, Hun-Ku
    • Journal of Life Science
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    • v.20 no.5
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    • pp.676-682
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    • 2010
  • This study focused on typing of the extended-spectrum $\beta$-lactamase (ESBL) produced in organisms isolated from a natural environment, rather than a clinical setting. Samples were collected from drain water issuing from a sewage plant in Kwanganri (Busan, Korea). Following double disk synergy testing, 29 strains were selected as potential ESBL positive strains. Of these, 15 strains were transconjugants of the sodium azide resistant recipient strain Escherichia coli J53 and analyzed biochemically including indole, methyl-red, Voges-Proskauer, Simmon's citrate, decarboxylase-dihydrolase and sugar-fermentation tests. The tests classified the 15 strains as Klebsiella pneumoniae (n=13) and Escherichia coli (n=2). The type of ESBL from each strain was deduced by isoelectric focusing point analysis and DNA sequencing. The results indicated that the types of ESBL were SHV-12 (n=4) and SHV-12/TEM-1 (n=9) from K. pneumoniae and TEM-1 (n=2) from E. coli strains.

Preparation and Biocompatibility of 6-amino-6-deoxychitosan for Immobilization of Epidermal Growth Factor (세포성장인자 고정화를 위한 6-amino-6-deoxychitosan의 제조와 생체적합성)

  • Son, Tae Il;Park, Se Hoon;Kang, Hahk Soo;Jang, Eui Chan
    • Applied Chemistry for Engineering
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    • v.16 no.2
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    • pp.226-230
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    • 2005
  • Chitosan derivatives, 6-amino-6-deoxychitosan (6A6DC) was successively prepared as a reactive carbohydrate for the stabilization of epidermal growth factor (EGF) by the reactions of chitosan with tosyl chloride, sodium azide, and lithium aluminum tetrahydride. The structure of 6A6DC was confirmed by EA, FT-IR, $^1H-NMR$ and $^{13}C\{^1H\}-NMR$. The degree of substitution (ds) of amino groups in 6A6DC was determined to be 0.7. 6A6DC did not show any cytotoxicity on the normal human dermal fibroblast (NHDF) proliferation at least in the range tested (0.3 g/mL 600 g/mL) and was considered as a suitable material for the stabilization of EGF against proteolytic degradation due to its non-cytotoxicity and high reactivity.

A New Double-Petaled Pasqueflower (Pulsatilla koreana) Variety, 'Gyeobi' (겹꽃 분화용 할미꽃 신품종 '겨비')

  • Lee, Ya-Seong;Kim, Dong-Kwan;Choi, Duck-Soo;Choi, Jin-Kyung;Son, Dong-Mo;Choi, Kyeong-Ju;Baek, Hyeong-Jin;Rim, Yo-Sup
    • Korean Journal of Breeding Science
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    • v.42 no.3
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    • pp.245-247
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    • 2010
  • A new pasqueflower (Pulsatilla koreana) variety, 'Gyeobi,' was derived from a mutation of native P. koreana at the Jeollanamdo Agricultural Research and Extension Services (JARES) in Naju, Korea. The 'Gyoebi' was obtained by treating seeds with 3 mM of a chemical mutagen, sodium azide, for 16 hr in 2001. The variety was established in 2005 after two years of selective breeding. 'Gyeobi' is characterized by reddish purple flowers with 12 double petals. The inherent characteristics of the variety are deep yellow anthers and reddish purple stigmas. The agronomic characteristics of the variety are 6.3 flowers per plant, 26.4 cm in flower height, 14.2 cm in leaf length, 7.5 cm in flower width, and 11.0 cm in bract width.

Effect of Ascorbic Acid on the Activities of Ethanol Metabolizing Enzymes (Ascorbic acid가 에탄올 대사효소에 미치는 영향)

  • Kim Yong-Sik
    • The Korean Journal of Pharmacology
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    • v.20 no.1 s.34
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    • pp.47-54
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    • 1984
  • Effect of ascorbic acid on various hepatic ethanol metabolizing enzymes including alcohol dehydrogenase(ADH), the microsomal . ethanol oxidizing system(MEOS), and catalase was quantitatively evaluated in liver microsomal and cytosolic preparation from Sprague-Dowley rats. In present study, ADH activities were no changed significantly by ascorbic acid. The MEOS activity, dependent on NADPH and $O_2$, was affected by azide (inhibitor of catalase) or exogenous catalase. In the presence of ascorbic acid, ethanol oxidation by rat liver microsomal preparation reacted with NADPH-generating system was increased by up to 22.5%, but decreased when liver microsome was reacted with $H_2O_2$ generated by xanthine and xanthine oxidase. Increase in the activity of the MEOS in the presence of ascorbic acid was greater in liver microsomal preparation pretreated with azide. Also ascorbic acid oxidized ethanol nonenzymatically. This ethanol oxidation induced by ascorbic acid was inhibited by OH radical scavengers (thiourea, sodium benzoate), but was not much affected by superoxide dismutase. From these results it was suggested that ascorbic acidcould interact directly with the MEOS, then promote the oxidation of ethanol. And, to some extent, ${\cdot}OH$-radicals or other radicals generated during the spontaneous autooxidation of ascorbic acid may be responsible for the production of acetaldehyde from ethanol.

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Study of Cytotoxicity of an Actinomycete Isolated in Korea (토양에서 분리한 방선균의 세포 독성에 관한 연구)

  • Park, Joon-Koo;Choi, Boung-Don;Kim, Seung-Chul;Ryeom, Kon
    • Environmental Analysis Health and Toxicology
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    • v.8 no.3_4
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    • pp.7-12
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    • 1993
  • An Actinomycete strain isolated from Mt. Dea-Dun had a strong antifungal activity. The culture brith produced by isolated strain showed only antifungal activity against fungi with the exception of yeast and bacteria. It was heat stable, dissolved in ehtylacetate. The concentrated antifungal agent showed cytotoxicity against HEP-2 and HeLa as tumor cell line, and showed weak cytotoxicity against VERO 36 as normal cell line. Morphological and physiological characteristics were tested with isolated strain. The spore color of isolated strain was gray. It had a short chain and produced brown colored lytic substance in yeast extract-malt agar. The cell wall of isolated strain was composed of meso-DAP, and we suggested it as genus Actinomadura. In the existing of chemical inhibitor, isolated strain grew on the condition of 0.0001% crystal violet, 0.1% phenol, 0.01% sodium azide and 10% sodium chloride. Carbon utilization of isolated strain was shown that glucose, sucrose, manitol and sodium citrate were well utilized.

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A New Selective Medium for the Isolation and the Detection of Leuconostocs in Foodstuffs (식품중에 함유된 Leuconostocs 균주의 새로운 선택배지 개발)

  • Choi, Hak-Jong;Shin, Young-Jae;Yu, Ju-Hyun;Yoon, Sung-Sik
    • Korean Journal of Food Science and Technology
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    • v.28 no.2
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    • pp.279-284
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    • 1996
  • To develop a selective medium for the isolation and the detection of leuconostocs from the various samples including fermented vegetables, ten strains of leuconostocs and seven strains of lactobacilli were tested for their sensitivity to various antibiotics. The basal-medium containing 5 ${\mu}g/ml$ of novobiocin inhibited the growth of lactobacilli completely, but not that of leuconostocs. On the basis of this result, a new selective medium was developed and to be named NLS medium. This medium contains 1% Tryptone (Difco), 0.1% Yeast Extract (Difco), 2% sucrose, 0.1% Beef Extract (BBL), 0.5% sodium acetate, 0.2% ammonium sulfate, 0.01% magnesium sulfate, 0.2% dipotassium phosphate, 0.05% sorbic acid, 75 ppm sodium azide (Sigma), 0.1% (vol/vol) Tween 80, 30 ${\mu}g/ml$ of Vancomycin (Sigma), 5${\mu}g/ml$ of Novobiocin (Sigma), 0.5${\mu}g/ml$ of cysteine HCI, and 1.5% Agar (Difco). All of the eighty six isolates obtained from some foodstuffs were identified as members of the genus Leuconostoc. Comparative counts with the MRS, PES, LUSM, and NLS medium indicated that the recovery percent was lower than other selective media. Therefore, this result suggested that NLS medium was suitable for the isolation of leuconostocs, but not for counting or enumerating.

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Analysis of Antioxidative Activity and Antimutagenic Effect of Ethanol Extract from Schizandra chinensis Baillon (오미자(Schizandra chinensis Baillon) 에탄올 추출물의 항산화 활성 및 항돌연변이 활성 분석)

  • Jeon, Yeon-Bee;Kil, Jin-Bee;Lim, So-Min;Kim, Mee-Hyun;Kim, Mee-Ra
    • Journal of the East Asian Society of Dietary Life
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    • v.18 no.5
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    • pp.746-752
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    • 2008
  • In this study, the antioxidative activity and antimutagenic effect of ethanol extracts from Schizandra Chinensis Baillon were assessed with regard to natural antioxidants. The antioxidative activity and antimutagenicity of ethanol extracts from Schizandra chinensis Baillon were evaluated by measuring the radical-scavenging effect on 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals, and by performing the Ames test using 4-nitroquinoline-1-oxide (4-NQO) and sodium azaid as a mutagen with Salmonella typhimurium TA100, respectively. The total polyphenolic compound and flavonoid compound contents of Schisandra chinensis Baillon were also assessed. The DPPH radical-scavenging activity of ethanol extracts from Schisandra chinensis Baillon was 57.2% on the $500\;{\mu}g$/assay. The $IC_{50}$ on DPPH radical-scavenging effect of ethanol extract was $435\;{\mu}g$/assay. In addition, the inhibition rates of ethanol extract from Schisandra chinensis Baillon toward mutagenicity induced by 4-NQO and sodium azide were 82.45% and 45.3%, respectively. Furthermore, the total polyphenol and total flavonoid contents of the extract were 9.53 mg/g and 3.97 mg/g, respectively. These results indicate that the ethanol extract from Schisandra chinensis Baillon evidences a fairly good antioxidative and antimutagenic effect.

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Purification and Characterization of Laccase from Basidiomycete Fomitella fraxinea

  • Park, Kyung-Mi;Park, Sang-Shin
    • Journal of Microbiology and Biotechnology
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    • v.18 no.4
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    • pp.670-675
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    • 2008
  • A laccase was isolated from the culture filtrate of the basidiomycete Fomitella fraxinea. The enzyme was purified to electrophoretical homogeneity using ammonium sulfate precipitation, anion-exchange chromatography, and gel-filtration chromatography. The enzyme was identified as a monomeric protein with a molecular mass of 47 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and gel-filtration chromatography, and had an isoelectric point of 3.8. The N-terminal amino acid sequence for the enzyme was ATXSNXKTLAAD, which had a very low similarity to the sequences previously reported for laccases from other basidiomycetes. The optimum pH and temperature for 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonate) (ABTS) were 3.0 and $70^{\circ}C$, respectively. The enzyme also showed a much higher level of specific activity for ABTS and 2,6-dimethoxyphenol (DMP), where the $K_m$ values of the enzyme for ABTS and 2,6-DMP were 270 and $426{\mu}M$, respectively, and the $V_{max}$ values were 876 and $433.3{\mu}M/min$, respectively. The laccase activity was completely inhibited by L-cysteine, dithiothreitol (DTT), and sodium azide, significantly inhibited by $Ni^+,\;Mn^{2+}$, and $Ba^{2+}$, and slightly stimulated by $K^+$ and $Ca^{2+}$.