• Title/Summary/Keyword: acp

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Molecular Analysis of Isoniazid-Resistance Related Genes of Mycobacterium tuberculosis Isolated from Korea

  • Hwang Joo Hwan;Jeong Eun Young;Choi Yeon Im;Bae Kiho;Song Taek Sun;Cho Sang-Nae;Lee Hyeyoung
    • Biomedical Science Letters
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    • v.11 no.4
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    • pp.455-463
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    • 2005
  • Resistance to isoniazid (INH), which is one of the most important drugs in Mycobacterium tuberculosis chemotherapy, has been associated with mutations in genes encoding the mycobacterial catalse-peroxidase (katG), the enoyl acyl carrier protein (ACP) reductase (inhA), alkyl hydroperoxide reductase (ahpC), beta-ketoacyl acyl carrier protein synthase (kasA), and NADH dehydrogenase (ndh). In this study, we examined INH-resistance related genes in 50 INH-resistant and 24 INH-susceptible isolates by PCR-sequence analysis. In brief, mutations at the katG gene were found at codon 315 alone (2/50), at codon 463 alone (19/50), and both at 315 and 463 (29/50). However, while mutations at codon 315 were only detected in INH-resistant isolates, mutations at codon 463 were also detected in INH-susceptible isolates indicating mutations at 463 alone do not seem to confer resistance to INH. Similar to the case of katG 463, some of inhA mutations were also found among INH-susceptible isolates. For example, whereas mutations at 8 upstream of the start codon (UPS) and 15 UPS of the inhA gene were detected only in INH-resistant isolates, mutations at 101, 115, and 125 UPS were detected only in INH-susceptible isolates. Many different kinds of mutations were detected in INH­resistant isolates at ahpC, oxyR gene, and intergenic region of the oxyR-ahpC genes. Howerver, the mutations were not found oxyR and the intergenic regions in INH-susceptible isolates. No mutations were found at either kasA or at ndh gene among INH-resistant isolates. In conclusion, some of mutations such as katG 315, inhA promotor region, and oxyR-ahpC seem to be strongly related to INH-resistance. Currently we are developing a molecular diagnostic method based on these results.

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Ethanol Production an Immobilized Themotolerant Mutant of Brettanomyces custersii H1-39 from Wood Hydrolyzate Media (목질계 당화액배지로부터 고온내성 변이주 Brettanomyces custersii H1-39의 고정화에 의한 에탄올생산)

  • 박승원;홍영기;김승욱;홍석인
    • Microbiology and Biotechnology Letters
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    • v.28 no.3
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    • pp.172-179
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    • 2000
  • Bretlanomyces C!tstersii Hl-39 mutant was immobilized with various caniers. Immobilized mutant Hl-39 produced more ethanol and showed higher productivity and cell concentration than those of free 81-39 in 3.4% hydrolyzate of wood-chips at different temperatures ($37^{\circ}C$, $40^{\circ}C$ and $43^{\circ}C$). At $37^{\circ}C$, ethanol concentration produced by mutant H1-39 immobilized in Ca-alginate and ARG(l % Ca-alginate, 1.67% bentonite, 0.33% glutaraldehyde) bead were higher than those produced by the other earners (ACG ; 1 % CaHalginate. ] .67% celite R-634 , 0.33% glutaraldehyde, ABP ; 1 % Ca-alginate. 1.67% bentonite, 0.33% pectin. ACP: 1 % Ca-alginate, ] .67% celiLe R-634, 0.33% pecLin). The highest value of productivity(l.23 ) was obtained by using ABG beads. At $40^{\circ}C$, ethanol conccntration and productivity obtained by ABC beads ,>,"ere 15.2 glL and 0.84 gl L.h, respectively, which showed the highest value compared to other carriers. Particularly, productivity of ilmnobilized ceIl was increased up to 90% as compared to that offree cell. On the other hand, ABP(l % Ca-alginate+L67% bentonile+O.33% pectin) beads gave the best resulLs at $43^{\circ}C$ for production of ethanol and productivity, which were 13.8 g!l and 0.77 g/l h, respectively.ively.

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Action modes of the anti-complementary polysaccharides purified from Arecae pericarpium (대복피로부터 정제된 보체활성화 다당의 작용양식)

  • Shin, Kwang-Soon;Cho, Hong-Yon;Sung, Ha-Chin;Yang, Han-Chul
    • Applied Biological Chemistry
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    • v.35 no.6
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    • pp.462-469
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    • 1992
  • Two kinds of complement activating (anti-complementary) polysaccharides, which were expected to be immunomodulators were purified from Arecae Pericarpium (the pericarps of Areca catechu), and their action modes have been studied. The active polysaccharides, AC-2-IIIa and AC-2-IIIc from Arecae Pericarpium showed dose-dependent anti-complementary activities on $TCH_{50}$. The anti-complementary activities of AC-2-IIIa and AC-2-IIIc in metal ion-free condition were completely decreased in comparison with control whereas in case of $Ca^{2+}$-free condition, these activities were maintained, considerably. Also AC-2-IIIa and AC-2-IIIc showed relatively potent alternative complement pathway activities. Furthermore, after incubation of the normal human serum with polysaccharide of Arecae Pericarpium in the absence of $Ca^{2+}$ ion, a cleavage of C3 in the serum was found to have occurred through immunoelectrophoresis (IEP) with anti-human C3. Also, from the results of IEP using anti-human whole serum, the ratios of the height of 3rd peak to ${\alpha}2-M$ peak by AC-2-IIIa and AC-2-IIIc proved to be $1.50{\pm}0.04$ and $1.22{\pm}0.08$, respectively. These results indicate that the modes of complement activation by AC-2-IIIa and AC-2-IIIc from Arecae Pericarpium are via both the classical pathway and the alternative pathway.

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Identification of Differentially Expressed Genes in Four Different Growing Stages in Korea Native Chicken Liver (황갈색 재래닭의 간에서 성장 단계별 차등 발현 유전자 분석)

  • Lee, K.Y.;Yu, S.L.;Jung, K.C.;Jang, B.K.;Choi, K.D.;Lee, J.H.
    • Korean Journal of Poultry Science
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    • v.34 no.2
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    • pp.85-90
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    • 2007
  • The chicken liver has been involved in various biological functions including detoxification, glycogen storage and plasma protein synthesis. The aim of this study was to investigate differentially expressed genes in chicken liver in four different growing stages. Using 10 arbitrary Annealing Control Primers (ACPs), five differentially expressed genes have been identified. Based on the Basic Local Alignment Search Tool (BLAST) search results, three of them were matched with previously known genes, and the other two were matched with unknown EST sequence and a hypothetical protein, respectively. In order to confirm the expression results, quantitative real-time PCR was also performed. The high similarities between the expression data using arbitrary ACPs and quantitative real-time PCR indicate that the identified genes are the real differentially expressed genes in different growing stages. The genes identified in this study can be used as valuable biomarkers in chicken with further investigation of the functions.

Effect of p-Phenylenediamine to Rat Skin (p-Phenylenediamine이 흰쥐의 피부 조직에 미치는 영향)

  • Lee, SangHee;Cho, HyunGug;Lee, Sang-Il
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.9
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    • pp.1330-1335
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    • 2005
  • Red brownish p-pheylenediamine (PPD) has been widely used hair dye for women. The dye was known to cause systemic anaphylaxis, dermatitis and bladder cancer. But the effect of PPD toxicity with oxygen free radical has not been studied. This study investigated the degree of skin injury by PPD. PPD ($2.5\%$ PPD in $2\%\;NH_{4}OH$) was applied to the rat skin ($25 mg/16.5\;cm^2$) 3 or 5 times every other day. Histopathological findings demonstrated the proliferation of epithelial cells and the increased keratinization by PPD. The activities of glucose 6-phosphatase (G6Pase) was decreased and acid phosphatase (ACP) was increased in PPD-applied rat skin. Groups in which PPD was applied 5 times were more damaged than groups applied 3 times. To examine the relationship between tissue damage and oxygen free radicals, effect of PPD on xanthine oxidase (XO) activity was measured and XO activity was more significantly increased in the group treated with PPD 5 times than 3 times. However, reduced glutathione (GSH) content, and the activities of catalase (CAT), super-oxide dismutase (SOD) and glutathione S -transferase (GST) were more decreased in PPD-applied groups than in controls. Even though the activities of XOD was not changed in the group treated with PPD 3 times, the decreased activities of oxygen free radical system and the damaged skin tissue were observed. This result might be caused by the production of toxic PPD metabolites in rat skin. In conclusion, topical PPD application led to skin injury in a dose-dependent manner, probably due to the generation rate of oxygen free radical.

A Study on the Electrochemical Properties of Porous Carbon Electrode according to the Organic Solvent Contents (유기용매의 함량비에 따른 다공성 탄소전극의 전기화학적 특성 연구)

  • Lim, Jung-Ae;Choi, Jae-Hwan
    • Applied Chemistry for Engineering
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    • v.19 no.2
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    • pp.185-190
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    • 2008
  • In order to increase the surface area of electrodes for electrosorption, porous carbon electrodes were fabricated by a wet phase inversion method. A carbon slurry consisting of a mixture of activated carbon powder (ACP), polyvinylidene fluoride (PVdF), and N-methyl-2-pyrrolidone (NMP) as a solvent was cast directly on a graphite sheet. The cast film was then immersed in pure water for phase inversion. The physical and electrochemical properties of the electrodes were investigated using scanning electron microscopy (SEM), porosimetry, and cyclic voltammetry. The SEM images verified that the pores of various sizes were formed uniformly on the electrode surface. The average pore sizes determined for the electrodes fabricated with various NMP contents ranged from 64.2 to 82.4 nm and the size increased as the NMP content increased. All of the voltammograms showed a typical behavior of charging and discharging characteristic at the electric double layer. The electrical capacitance ranged from 3.88 to $5.87F/cm^2$ depending on the NMP contents, and the electrical capacitance increased as the solvent content decreased. The experimental results showed that the solvent content is an important variable controlling pore size and ultimately the capacitance of the electrode.

Effects of Salt and Drought Stresses on Seed Germination and Gene Expression Pattern in Tall Fescue (염과 건조 스트레스 조건에서 톨 페스큐의 종자 발아율과 유전자 발현 변화분석)

  • Lee, Sang-Hoon;Lee, Ki-Won;Choi, Gi Jun;Kim, Ki-Yong;Ji, Hee Jung;Hwang, Tae Young;Lee, Dong-Gi
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.34 no.2
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    • pp.114-119
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    • 2014
  • Salinity and drought stresses are probably the most significant abiotic factor limiting plant's growth, also negatively affect seed germination and early seedling development. To study on effect of NaCl and PEG stress on seed germination and gene expression pattern of tall fescue, the levels of NaCl and PEG-induced water stresses were determined in first experiment. Different concentration of NaCl (0 to 350 mM) and PEG (0 to 30%) were used for seed treatment. Seed Germination percentage reduced with increasing osmotic potential of growth medium either due to NaCl or PEG. Seeds were not germinate at 350 mM NaCl or 30% PEG treatment. On the basis of the results, Kentucky31(E-) had more resistant than Fawn in both stress conditions. Furthermore, we have used an annealing control primer-based differential display reverse transcription-polymerase chain reaction method to identify salt- and drought stress-induced differentially expressed genes (DEGs) in tall fescue leaves. Using 120 annealing control primers, a total of 4 genes were identified and sequenced. The possible roles of the identified DEGs are discussed in the context of their putative role during salinity and drought stresses.

Chitosan and N-acetylchitosan film from Squid Pen and Their Characteristics (오징어 연골을 이용한 Chitosan 및 N-acetylchitosan film의 제조 및 특성)

  • CHOI Hyeon-Mee;LEE Keun-Tai
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.33 no.4
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    • pp.356-360
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    • 2000
  • In order to utilize the processing wastes of squid, chitosan was prepared by intermittent deacetylation treatment of ${\beta}-chitin$ richly contained in the pen of squid, and then their characteristics of chitosan and N-acetylchitosan film were studied. The acetylation time of N-acetylchitosan film-1 (N-ACE-1) manufactured from chitosan solution by treating with acetic anhydride was about 12 hrs. In SEM photomicrographs, the surface of chitosan film was regularly arranged netlike, and that of N-acetylchitosan film-2 (N-ACE-2) was rough like snowflake and larger than chitosan film. The chitosan film (thickness 0.02 mm, time 60 min) had the highest tensile strength ($1,240 kg/cm^2$) and elongation ($58.25{\%}$), N-ACP-1 (thickness 0.02 mm, time 60 min) had the highest water permeability ($539 g/m^2{\cdot}24 hrs$), oxygen permeability ($20,000 cm^3/m^2{\cdot}24 hrs{\cdot}atm$) and water uptake ($350{\%}$) among the tested films.

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Identification of intrinsic regulators in the secondary palate morphogenesis (이차구개 형태분화의 내적 조절유전자 규명)

  • Lee, Jae-Guk;Jang, Eun-Ha;Im, Yang-Hee;Kim, Ki-Byeung;Ko, Seung-O;Cho, Eui-Sic;Shin, Hyo-Keun
    • Korean Journal of Cleft Lip And Palate
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    • v.10 no.1
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    • pp.1-16
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    • 2007
  • 이차구개는 발생과정에서 구개선반의 형성과 성장, 거상과 융합의 과정을 통해 형성된다. 이와 같은 이차구개의 형성과정은 미세한 분자유전학적 신호전달기전에 의해 조절되는 것으로 알려져 있어서, 신호전달과정에 관여하는 유전인자의 발현이상이 되면 정상적인 이차구개가 형성되지 못하고 구개파열이라는 선천성 기형이 발생된다. 구개파열의 유발인자들에 대한 많은 연구에도 불구하고 현재까지 정상적인 이차구개의 형성을 조절하는 분자유전학적 기전에 대해서는 명확히 알려져 있지 않다. 따라서 본 연구에서는 이차구개의 형태분화를 조절하는 분자유전학적 기전을 알아보고자, 이차구개 형성의 내적 조절인자 중 핵심유전자로 알려져 있는 Osr2가 결손된 생쥐의 이차구개 형성과정에서 정상생쥐에 비해 발현의 변동이 나타나는 유전자를 확인하였다. 유전자 발현의 변동은 발생 14.5일(E14.5)의 구개선반으로부터 추출한 total RNA를 이용하여 ACP-based GeneFishing PCR을 시행하여 확인하였고, 각각의 변동된 유전자를 동정하여 정상생쥐의 이차구개 형성과정에서의 발현양상을 in situ hybridization을 시행하여 확인하였다. 총 120쌍의 primer를 이용한 검색을 통해서 정상생쥐의 구개선반에 비해 mutant에서 발현이 변동된 유전자는 7개가 검출되었고, 이들은 모두 정상생쥐에 비해 mutant에서 발현이 증가되는 것으로 확인되었다. 검출된 유전자는 vimentin(Vim), ${\beta}$-tropomyosin 2(Tpm2), thioredoxin-like 5(Txnl5), procollagen type II alpha 1(Col2a1), Insulin-like growth factor binding protein 7(IGFbp7), Sui 1 homologs(Sui 1), Defender against cell death1(Dad1)이었다. 검출된 유전자를 동정하여 정상생쥐의 구개 형성과정에서의 발현양상을 알아본 결과, Col2a1 을 제외한 유전자들은 모두 E13.5의 구개선반에서 특이적으로 발현되고 있었으나 구개선반이 융합된 E15.5에서는 Vim, Txnl5 그리고 Dad1 만이 봉합선을 따라 발현이 지속되고 있었다. 이상의 결과로 보아 검출된 유전자들은 구개선반의 형태분화과정에서 발현되어 이차구개의 형성과정에 관여할 것으로 여겨진다. 또한 이들은 이차구개 형성의 내적조절인자인 Osr2의 downstream target 으로 구개선반의 성장과 융합과정에 직접적으로 관여하는 유전물질일 것으로 추정된다.

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Effects of Dietary Calcium and Soy Isoflavones Supplementation on Bone Metabolism in the Ovariectomized Rats (난소절제 흰쥐의 골격대사에 대한 식이 칼슘과 대두 이소플라본의 섭취효과)

  • Kim, Min-Sun;Lee, Yeon-Sook
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.6
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    • pp.833-839
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    • 2005
  • This study was conducted to demonstrate whether dietary Ca and the soy isoflavones supplementation could reduce the bone loss associated with estrogen deficiency. Nine week-old female rats (SD) were ovariectomized and then fed on diet of low $(0.1\%)$ or normal $(0.5\%)$ Ca supplemented with soy isoflavones (80 and 160 ppm) for 6 weeks. The ovariectomized and sham-operated rats showed normal serum Ca and P levels, and dietary Ca and soy isoflavones did not changed them. The serum alkaline phosphatase activity was increased in all ovariectomized rats, especially in the rats fed low Ca diet regardless of isoflavone supplementation. The serum tartrate-resistance acid phosphatase was not significantly different among the ovariectomized rats and were not changed by dietary Ca and isoflavones. Breaking force of femur was higher in the rats fed the nomral Ca diets and not statistically changed by soy isoflavone supplementation. Femoral and lumbar Ca and P contents decreased in the ovariectomized rats and the soy isoflavones-80 ppm supplementation significantly enhanced bone minerals, but the soy isoflavones-160 ppm supplementation did not. Dietary Ca increased lumbar Ca and P contents. The results of this study have suggested that the soy isoflavones 80 ppm supplementation could be sufficient to prevent bone loss in ovariectomized rats and normal Ca supplementation could enhance the effect of soy isoflavones on bone protection.