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Toxin Produced by Pathogenic Vibrios Isolated from Sea Food (수산물에서 분리된 병원성 비브리오균의 용혈성독소)

  • CHANG Dong-Suck;SHINODA Sumio
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.27 no.2
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    • pp.107-113
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    • 1994
  • Among the currently recognized pathogenic vibrios, V. vulnificus and V. cholerae non O1 are the most serious bacteria from the point of view of sea food hygiene in Korea. In this paper, the authors compared the hemolytic activities of the crude hemolysin produced by V. vulnificus and V. cholerae non O1 isolated from shellfish collected in Chungmoo, Korea. The authors also attempted to improve the purification method of V. vulnificus hemolysin(VVH) and tried to make antiserum with the purified hemolysin. VVH was produced in abundance in heart infusion broth containing $2\%$ NaCl in a shaking cultivation process(140rpm) at $37^{\circ}C$ for 15 hours. While hemolysin production patterns of V. cholerae non O1 were quite different by the strain during the culture times compared with the V. vulnificus. Hemolytic activity of the VVH on sheep erythrocytes was stronger than those of rabbit, but hemolytic activities of the hemolysin produced by V. cholerae non O1 on rabbit erythrocytes were as much as twice as strong as on those of sheep and horse. VVH was purified by two steps of hydrophobic column chromatography on Phenyl-Sepharose HP with Fast Protein Liquid Chromatography(FPLC). Purification fold and yield of VVH was much improved by changing the elution buffer's pH from 6.0 to 9.8 and adding $1\%$ CHAPS(a zwitter ionic detergent) and $50\%$ ethylene glycol to the 10mM glycine buffer during the repeated hydrophobic column chromatography. Homogeneity of the purified hemolysin was shown by polyacrylamide gel electrophoresis. According to the five times repeated purification results, the specific activity was increased 27500 times and the yield was improved by $23.4\%$ on average. About $250{\mu}g$ of purified hemolysin was harvested from the 2400ml of culture supernatant of V. vulnificus. Molecular weight of VVH was estimated to be 50KDa by the SDS-PAGE and the neutralization scores of the obtained antiserum acting against VVH were $2000{\sim}8500$.

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Feasibility Study on Double Path Capacitive Deionization Process for Advanced Wastewater Treatment (이단유로 축전식 탈염공정의 하수고도처리 적용가능성 평가)

  • Cha, Jaehwan;Shin, Kyung-Sook;Lee, Jung-Chul;Park, Seung-Kook;Park, Nam-Su;Song, Eui-Yeol
    • Journal of Korean Society of Environmental Engineers
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    • v.36 no.4
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    • pp.295-302
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    • 2014
  • This study demonstrates a double-path CDI as an alternative of advanced wastewater treatment process. While the CDI typically consists of many pairs of electrodes connected in parallel, the new double-path CDI is designed to have series flow path by dividing the module into two stages. The CFD model showed that the double-path had uniform flow distribution with higher velocity and less dead zone compared with the single-path. However, the double-path was predicted to have higher pressure drop(0.7 bar) compared the single-path (0.4 bar). From the unit cell test, the highest TDS removal efficiencies of single- and double-path were up to 88% and 91%, respectively. The rate of increase in pressure drop with an increase of flow rate was higher in double-path than single-path. At 70 mL/min of flow rate, the pressure drop of double-path was 1.67 bar, which was two times higher than single-path. When the electrode spacing was increased from 100 to $200{\mu}m$, the pressure drop of double-path decreased from 1.67 to 0.87 bar, while there was little difference in TDS removal. When proto type double-path CDI was operated using sewage water, TDS, $NH_4{^+}$-N, $NO_3{^-}$-N and $PO_4{^{3-}}$-P removal efficiencies were up to 78%, 50%, 93% and 50%, respectively.

Bee Venom Inhibits Angiogenesis by Decreasing HIF-1α Expression in HCT116 Cells (봉독의 HIF-1α 발현감소를 통한 혈관신생 억제효과)

  • Shin, Jae-Moon;Jeong, Yun-Jeong;Park, Kwan-Kyu;Choe, Jung-Yoon;Han, Sang-Mi;Lee, Kwang-Gill;Yeo, Joo-Hong;Chung, Il-Kyung;Chang, Young-Chae
    • Journal of Life Science
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    • v.22 no.1
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    • pp.41-48
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    • 2012
  • Bee venom (BV) has been used in medicine to treat a variety of diseases including arthritis, rheumatism, and various cancers. Recent reports indicate that BV has anti-angiogenic effects, but the precise molecular mechanism underlying the effects of BV against colorectal cancer remains to be elucidated. We examined the effects of BV and its major components (melittin and apamin) on tumor angiogenesis and found that BV significantly decreased protein levels of hypoxia-inducible factor-$1{\alpha}$ (HIF-$1{\alpha}$), an important factor involved in angiogenesis and tumor progression, in human colorectal carcinoma HCT116 cells. BV also suppressed the transcription of HIF-$1{\alpha}$ under hypoxia, leading to a decrease in the expression of vascular endothelial growth factor (VEGF), a major target gene of HIF-$1{\alpha}$. We also found that these effects were mainly elicited by apamin, but not melittin. BV specifically inhibited the phosphorylation of ERK1/2 without changing the total levels of this protein, but had no effect on kinases of p38/JNK and AKT. Our results suggest that BV may inhibit human colorectal cancer progression and angiogenesis by inhibiting HIF-$1{\alpha}$ and VEGF expression, thereby providing a novel potential mechanism for the anticancer action of BV.

Superovulation Response by FSH Treatments in Hanwoo (FSH 투여 용량과 방법Ul 따른 한우의 과배란 처리 효율)

  • Choi, Soo-Ho;Park, Yong-Soo;Son, Woo-Jin;Lee, Joon-Hee;Rho, Gyu-Jin;Kim, Joo-Heon;Choe, Sang-Yong
    • Reproductive and Developmental Biology
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    • v.31 no.3
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    • pp.199-205
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    • 2007
  • The present study was carried out to assess the effect of superovulation response and efficiency of embryo production induced with injection of FSH dissolved in polyethylene glycol (PEG) in Hanwoo. Eighty-eight cows were divided into four groups. In group 1, cattle were intramuscularly treated with twice-daily administration of 50mg FSH for 4 days. Group 2 and 3 were subcutaneously single injection of 400mg and 200mg FSH dissolved in 30% PEG, respectively. Group 4 were subcutaneously single injection of 200 mg FSH dissolved in 30% PEG at 7 day after CIDR insertion. The number of corpus luteum (CL) in group 2 resulted in significantly (p<0.05) higher compared to group 1, 3 and 4 (18.5 vs. 11.2, 13.1 and 13.9, respectively). However, the number of total ova $(7.9{\sim}10.4)$, transferable embryos $(3.7{\sim}4.7)$, degenerate embryos $(1.9{\sim}3.5)$ and unfertilized ova $(1.8{\sim}2.7)$ did not differ among treatment groups. No difference was observed in pregnancy rate after transferring the recovered embryos among groups $(36.0{\sim}50.0%)$. In addition, blood progesterone concentrations at embryo recovery did not differ among all groups. In conclusion, although no differences were observed in the number of total ova, transferable embryos and pregnancy rate after transfer, a single injection of reduced dose of FSH (200mg FSH) at 7 day after CIDR insertion is more practical for superovulation treatments than frequent injection because of reduction of stress in Hanwoo and decreases of cost and laber.

Salinity Effects on Growth and Yield Components of Rice (관개용수내 염분농도가 벼 생육 및 수량에 미치는 영향)

  • Choi, Sun-Hwa;Kim, Ho-Il;Ahn, Yeul;Jang, Jeon-Ryeol;Oh, Jong-Min
    • Korean Journal of Ecology and Environment
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    • v.37 no.2 s.107
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    • pp.248-254
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    • 2004
  • This study was conducted to investigate the effects of salinity in irrigation water on the growth, yield components, yield and grain quality of rice plant by the pot experiments. Irrigation waters were supplied with control and amended with NaCl at 1,000, 2,000, 3,000, 5,000, and 7,000 ${\mu}s\;cm^{-1}$ electrical conductivity. A randomized block design was used with four replicates for each treatment and control. As increasing salt concentration, plant height, tiller number, SPAD value, dry weight, content of N, P, and K, ripened grain ratio (%), 1,000 grain weight, and protein content (%) tended to decrease, especially, significant at 3,000 ${\mu}s\;cm^{-1}$ of salt level. Grain yield decreased significantly at all treatments. The percentage of head rice slightly tended to increase as the salt concentration due to the decrease of green kernel. The percentage of green kernel was significantly lower at 3,000 ${\mu}s\;cm^{-1}$ of salt level than the control.

Potential Antitumor $\alpha$-Methylene-$\gamma$-butyrolactone-Bearing Nucleic Acid Base. 3. Synthesis of $5^1$-Methyl-$5^1$-[(6-substituted-9H-purin-9-yl)methyl]-$2^1$-oxo-$3^1$-methylenetetrahydrofurans

  • Kim, Jack-C.;Kim, Si-Hwan;Kim, Ji-A;Choi, Soon-Kyu;Park, Won-Woo
    • Archives of Pharmacal Research
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    • v.21 no.4
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    • pp.458-464
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    • 1998
  • Search for a new $\alpha$-methylene-$\gamma$-butyrolactone-bearing 6-substituted purine as a potental antitumor agent has led to synthesize seven, hitherto unreported, $5^1$-Methyl-$5^1$-[(6-substituted-9H-purin-9-yl)methyl]-$2^1$-oxo-$3^1$- methylenetetrahydrofurans (H, Cl, l, $CH_3$, $NH_2$, SH, >C=O) (6a-g). These include $5^1$-Methyl-$5^1$-[(9H-purin-9-yl)methyll-$2^1$-oxo-$3^1$ -methylenetetrahydrofurans (6a), $5^1$-Methyl-$5^1$-[(6-chloro-9H-purin-9-yl)methyl]-$2^1$-oxo-$3^1$-methylenetetrahydr ofurans (6b), $5^1$-Methyl-$5^1$-[(6-chloro-9H-purin-9-yl) methyl]-$2^1$-oxo-$3^1$-methylenetetrahydrofurans (6c), $5^1$-Methyl-$5^1$-[(6-methyl-9H-purin-9-yl) methyl]-$2^1$-oxo-$3^1$-methylenetetrahydrofurans (6d), $5^1$-Methyl-$5^1$-[(9H-adenin-9-yl)methyll-$2^1$-oxo-$3^1$-methylenetetrahydrofurans (6e), $5^1$-Methyl-$5^1$-[(6-mercapto-9H-purin-9-yl) methyl]-$2^1$-oxo-$3^1$-methylenetetrahydrofurans (6f) and $5^1$-Methyl-$5^1$-[(9H-hypoxanthin-9-yl)methyll-$2^1$-oxo-$3^1$-methylenetetrahydrof urans (6g) which were made by the Reformatsky-type reaction of ethyl $\alpha$-(bromomethyl) acrylate with the corresponding (6-substituted-9H-purin-9-yl)-2-propanone intermediates (5a-g). These ketone intermediates 5a-g, 1-(9H-purin-9-yl)-2-propanone (5a), 1-(6-chloro-9H-purin-9-yl)-2-propanone (5b), 1-(6-iodo-9H-purin-9-yi)-2-propanone (5c), 1-(6-methyl-9H-purin-9-yl)-2-propanone (5d), 1-(9H-adenin-9-yl)-2-propanone (Se), 1-(6-mercapto-9H-purin-9-yl)-2-propanone (5f), and 1-(9H-hypoxanthin-9-yl)-2-propanone (5g) were directly obtained by the alkylation of the 6-substituted purine bases with the chloroacetone in the presence of $K_2$$CO_3$ (or NaH) under DMF (or DMSO). The preliminary in vitro cytotoxcity assay for the synthetic .alpha.-methylene-y-butyro-lactone compounds (6a-g) were determined against three cell lines (PM-3A, P-388, and K-562) and showed the moderate antitumor activity ($IC_50$ ranged from 1.4 to 4.3 $\mu\textrm{g}$/ml) with the compound $5^1$-methyl-$5^1$ -[(9H-hypoxanthin-9-yl)methyl]-$2^1$-oxo-$3^1$-methylenetetrahydrofuran (6g) showing the least antitumor activity.

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Expression of Anion Exchanger and CFTR in the Hepatocyte and Cholangiocytes in Bile Duct-Ligated Rat (담관 결찰 쥐의 간세포와 담관세포의 anion exchanger와 CFTR 발현)

  • Lee, Jae-Dong;Wang, Joon-Ho;Ki, Seung-Seog;Choe, Won-Hyeok;Park, Jae-Seung;Cho, Won-Kyoo;Park, Jung-Jun;Kim, Hong-Su
    • Journal of Life Science
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    • v.21 no.12
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    • pp.1772-1777
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    • 2011
  • Cystic fibrosis transmembrane conductance regulator (CFTR) gene and sodium-independent $Cl^-/HCO_3^-$ anion exchanger (AE) genes are expressed in a wide variety of mammalian tissues including cholangiocytes. They play an important role in the regulation of intracellular pH (pHi) as well as in transepithelial acid/base transport necessary for biliary bicarbonate secretion. The aim of this study was to examine the expression level of CFTR gene and AE genes (AE1, AE2 and AE3) in the cholangiocytes and the hepatocytes, and also measure AE2 gene expression level after bile duct ligation (BDL). As we previously described, isolated hepatocytes and cholangiocytes from the liver of normal and BDL rats were prepared and gene expression levels were measured by using RT-PCR. We found that AE1, AE2, and AE3 genes were expressed in both hepatocytes and cholangiocytes, but CFTR was only in cholangiocytes. AE2 gene expression level was higher in the BDL hepatocytes than normal hepatocytes, which was significantly different between two groups. AE2 gene expression level was lower in the BDL cholangiocytes than normal cholangiocytes. However, AE2 gene expression level in both hepatocytes and cholangiocytes were not changed with a longer duration of BDL. These results suggest that CFTR and AE2 may play an important role in the pathogenetic mechanism of biliary cholestatic liver disease.

Electrofusion of Yeast Cells and Their Genetic Analysis Using RAPD-PCR (효모세포의 전기융합 및 융합세포의 RAPD-PCR을 이용한 유전적 분석)

  • Kim, Seung;Kim, Jae-Sung;Sapkota, Kumar;Park, In-Sung;Cho, Moon-Gu;Park, Yeal;Chun, Hong-Sung;Choi, Bong-Suk;Park, Se-Eun;Choi, Han-Suk;Kim, Myung-Kon;Kim, Sung-Jun
    • Applied Biological Chemistry
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    • v.49 no.3
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    • pp.186-191
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    • 2006
  • In the present study, an attempt has been made to produce hybrid yeast strains of different useful and dominant characteristics. The hybrid yeast strains were produced by electrofusion and their genetic analysis were performed by RAPD-PCR (random amplified polymorphic DNA-polymerase chain reaction). The protoplast of Saccharomyces cerevisiae KCTC 7904 and Zygosaccharomyces rouxii KCTC 7966 were obtained above 92% when treated with lyticase at $30^{\circ}C$ for $60{\sim}90$ min after the pretreatment of $1{\sim}2%$ 2-mercaptoethanol at $30^{\circ}C$ for $15{\sim}20$ min. The fusant was produced from paired protoplast stage under the electric pulse at high frequency conditions (1.5 MHz/50 pV, 615 $V/256\;{\mu}sec$) within glass-platinum made electrofusion chamber. Changes in RAPD patterns in mother cells and hybrid cells proved that the fusant contains two types of yeast gene originated from its parent. Furthermore, fermentation characters exhibits by the fusant cell confirmed its genetic changes. These results suggest that genetically stable hybrid yeast strains of economic importance can be produced by electrofusion technique and these electrofused yeast cells have an enormous impact in biotechnology and biomedicine.

THE COMPARISON OF DIFFERENT CANAL IRRIGATION METHODS TO PREVENT REACTION PRECIPITATE BETWEEN SODIUM HYPOCHLORITE AND CHLORHEXIDINE (차아염소산나트륨과 클로르헥시딘의 반응침전물 형성방지를 위한 여러 가지 근관세척 방법의 비교)

  • Choi, Moon-Sun;Park, Se-Hee;Cho, Kyung-Mo;Kim, Jin-Woo
    • Restorative Dentistry and Endodontics
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    • v.35 no.2
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    • pp.80-87
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    • 2010
  • The purpose of this study was to compare the different canal irrigation methods to prevent the formation of precipitate between sodium hypochlorite (NaOCl) and chlorhexidine (CHX). Extracted 50 human single-rooted teeth were used. The root canals were instrumented using NiTi rotary file (Profile .04/#40) with 2.5% NaOCl and 17% EDTA as irrigants. Teeth were randomly divided into four experimental groups and one control group as follows; Control group: 2.5% NaOCl only, Group 1: 2.5% NaOCl + 2% CHX, Group 2: 2.5% NaOCl + paper points + 2% CHX, Group 3: 2.5% NaOCl + preparation with one large sized-file + 2% CHX, Group 4: 2.5% NaOCl +95% alcohol+ 2% CHX. The teeth were split in bucco-lingual aspect and the specimens were observed using Field Emission Scanning Electron Microscope. The percentages of remaining debris and patent dentinal tubules were determined. Statistical analysis was performed with one-way analysis of variance (ANOVA). Energy Dispersive x-ray Spectroscopy was used for analyzing the occluded materials in dentinal tubule for elementary analysis. There were no significant differences in percentage of remaining debris and patent tubules between all experimental groups at all levels (p > .05). In elementary analysis, the most occluded materials in dentinal tubule were dentin debris. NaOCl/CHX precipitate was detected in one tooth specimen of Group 1. In conclusion, there were no significant precipitate on root canal, but suspected material was detected on Group 1. The irrigation system used in this study could be prevent the precipitate formation.

Prediction of Salvaged Myocardium in Patients with Acute Myocardial Infarction after Primary Percutaneous Coronary Angioplasty using early Thallium-201 Redistribution Myocardial Perfusion Imaging (급성심근경색증의 일차적 관동맥성형술 후 조기 Tl-201 재분포영상을 이용한 구조심근 예측)

  • Choi, Joon-Young;Yang, You-Jung;Choi, Seung-Jin;Yeo, Jeong-Seok;Park, Seong-Wook;Song, Jae-Kwan;Moon, Dae-Hyuk
    • The Korean Journal of Nuclear Medicine
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    • v.37 no.4
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    • pp.219-228
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    • 2003
  • Purpose: The amount of salvaged myocardium is an important prognostic factor in patients with acute myocardial infarction (MI). We investigated if early Tl-201 SPECT imaging could be used to predict the salvaged myocardium and functional recovery in acute MI after primary PTCA. Materials and Methods: In 36 patients with first acute MI treated with primary PTCA, serial echocardiography and Tl-201 SPECT imaging ($5.8{\pm}2.1$ days after PTDA) were performed. Regional wall motion and perfusion were quantified with on 16-segment myocardial model with 5-point and 4-point scaling system, respectively. Results: Wall motion was improved in 78 of the 212 dyssynergic segments on 1 month follow-up echocardiography and 97 on 7 months follow-up echocardiography, which were proved to be salvaged myocardium. The areas under receiver operating characteristic curves of Tl-201 perfusion score for detecting salvaged myocardial segments were 0.79 for 1 month follow-up and 0.83 for 7 months follow-up. The sensitivity and specificity of Tl-201 redistribution images with optimum cutoff of 40% of peak thallium activity for detecting salvaged myocardium were 84.6% and 55.2% for 1 month follow-up, and 87.6% and 64.3% for 7 months follow-up, respectively. There was a linear relationship between the percentage of peak thallium activity on early redistribution imaging and the likelihood of segmental functional improvement 7 months after reperfusion. Conclusion: Tl-201 myocardial perfusion SPECT imaging performed early within 10 days after reperfusion can be used to predict the salvaged myocardium and functional recovery with high sensitivity during the 7 months following primary PTCA in patients with acute MI.