• Title/Summary/Keyword: $S_N1$ mechanism

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Characterization of a Monoclonal Antibody Specific to Human Siah-1 Interacting Protein (인체 SIP 단백질에 특이적인 단일클론 항체의 특성)

  • Yoon, Sun Young;Joo, Jong Hyuck;Kim, Joo Heon;Kang, Ho Bum;Kim, Jin Sook;Lee, Younghee;Kwon, Do Hwan;Kim, Chang Nam;Choe, In Seong;Kim, Jae Wha
    • IMMUNE NETWORK
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    • v.4 no.1
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    • pp.23-30
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    • 2004
  • Background: A human orthologue of mouse S100A6-binding protein (CacyBP), Siah-1-interacting protein (SIP) had been shown to be a component of novel ubiquitinylation pathway regulating $\beta$-catenin degradation. The role of the protein seems to be important in cell proliferation and cancer evolution but the expression pattern of SIP in actively dividing cancer tissues has not been known. For the elucidation of the role of SIP protein in carcinogenesis, it is essential to produce monoclonal antibodies specific to the protein. Methods: cDNA sequence coding for ORF region of human SIP gene was amplified and cloned into an expression vector to produce His-tag fusion protein. Recombinant SIP protein and monoclonal antibody to the protein were produced. The N-terminal specificity of anti-SIP monoclonal antibody was conformed by immunoblot analysis and enzyme linked immunosorbent assay (ELISA). To study the relation between SIP and colon carcinogenesis, the presence of SIP protein in colon carcinoma tissues was visualized by immunostaining using the monoclonal antibody produced in this study. Results: His-tag-SIP (NSIP) recombinant protein was produced and purified. A monoclonal antibody (Korea patent pending; #2003-45296) to the protein was produced and employed to analyze the expression pattern of SIP in colon carcinoma tissues. Conclusion: The data suggested that anti-SIP monoclonal antibody produced here was valuable for the diagnosis of colon carcinoma and elucidation of the mechanism of colon carcinogenesis.

The Activity of c-Jun N -terminal Kinase (JNKb) in Patients with UIP (UIP 환자에서 c-Jun N-terminal Kinase (JNK) 활성화에 관한 연구)

  • Kim, Ki-Up;Lee, Young-Mok;Kim, Do-Jin;Moon, Seung-Hyuk;Uh, Soo-Taek;Kim, Yong-Hoon;Park, Choon-Sik;Kim, Hyun-Jo;Youm, Wook;Hwang, Jung-Hwa
    • Tuberculosis and Respiratory Diseases
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    • v.51 no.5
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    • pp.437-447
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    • 2001
  • Background: TNF-alpha is related to the generation of lung fibrosis in patients with UIP. The precise mechanism leading to lung fibrosis by TNF-alpha is unknown. However, the activation of a transcription factor like AP-1(down stream of c-jun N-terminal kinase, JNK) by TNF-alpha may be related to the induction of fibrogenic cytokines like PDGF or IGF-I. Furthermore, JNK was reported to be activated in the radiation-induced lung fibrosis model. This study examined JNK activity in patients with UIP. Methods : The expression of phosphorous JNK(p-JNK), macrophage/monocyte specific markers, CD68, and cytokeratin was evaluated by immunohistochemical(IHC) staining of lung tissues from patients with UIP and lung cancer. An in vitro kinase assay was performed with alveolar macrophages obtained by a bronchol-avleolar lavage from patients with UIP and healthy persons as the control. Results : The IHC stain showed that p-JNK is expressed in the almost all of the alveolar macrophages and smooth muscle cells in patients with UIP. In case of the normal areas of the lung from patients with lung cancer, the alveolar macrophages showed little p-JNK expression. Interestingly, increased JNK activity was not found in the in vitro kinase assay of the alveolar macrophages obtained from both patients with UIP and healthy persons as the control. Furthermore, 10 ng/mL of TNF-alpha failed to increase the JNK activity of the alveolar macrophages in both patients with UIP and healthy people. Conclusion : The JNK was activated constitutionally in patients with UIP. However, the role of JNK in the pathogenesis of lung fibrosis needs to be clarified.

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Nitric Oxide-mediated Relaxation by High $K^+$ in Human Gastric Longitudinal Smooth Muscle

  • Kim, Young-Chul;Choi, Woong;Yun, Hyo-Young;Sung, Ro-Hyun;Yoo, Ra-Young;Park, Seon-Mee;Yun, Sei-Jin;Kim, Mi-Jung;Song, Young-Jin;Xu, Wen-Xie;Lee, Sang-Jin
    • The Korean Journal of Physiology and Pharmacology
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    • v.15 no.6
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    • pp.405-413
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    • 2011
  • This study was designed to elucidate high-$K^+$ induced response of circular and longitudinal smooth muscle from human gastric corpus using isometric contraction. Contraction from circular and longitudinal muscle stripes of gastric corpus greater curvature and lesser curvature were compared. Circular smooth muscle from corpus greater curvature showed high $K^+$ (50 mM)-induced tonic contraction. On the contrary, however, longitudinal smooth muscle strips showed high $K^+$ (50 mM)-induced sustained relaxation. To find out the reason for the discrepancy we tested several relaxation mechanisms. Protein kinase blockers like KT5720, PKA inhibitor, and KT5823, PKG inhibitor, did not affect high $K^+$-induced relaxation. $K^+$ channel blockers like tetraethylammonium (TEA), apamin (APA), glibenclamide (Glib) and barium ($Ba^{2+}$) also had no effect. However, N(G)-nitro-L-arginine (L-NNA) and 1H-(1,2,4) oxadiazolo (4,3-A) quinoxalin-1-one (ODQ), an inhibitor of soluble guanylate cyclase (sGC) and 4-AP (4-aminopyridine), voltage-dependent $K^+$ channel (KV) blocker, inhibited high $K^+$ -induced relaxation, hence reversing to tonic contraction. High $K^+$-induced relaxation was observed in gastric corpus of human stomach, but only in the longitudinal muscles from greater curvature not lesser curvature. L-NNA, ODQ and KV channel blocker sensitive high $K^+$-induced relaxation in longitudinal muscle of higher portion of corpus was also observed. These results suggest that longitudinal smooth muscle from greater curvature of gastric corpus produced high $K^+$-induced relaxation which was activated by NO/sGC pathway and by $K_V$ channel dependent mechanism.

Neonatal Rat Necrotizing Enterocolitis Model Adopting Oral Endotoxin and Hypoxia Exhibits Increased Apoptosis through Caspase-3 Activation (경구 내독소와 저산소로 유발된 신생쥐의 괴사성 장염모델에서 caspase-3 활성화를 통한 세포자멸사의 증가)

  • Lee, Yun-Kyoung;Kim, Ee-Kyung;Kim, Ji-Eun;Kim, Yoon-Joo;Son, Se-Hyung;Kim, Han-Suk;Kim, Beyong-Il;Choi, Jung-Hwan
    • Neonatal Medicine
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    • v.17 no.1
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    • pp.44-52
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    • 2010
  • Purpose : The aim of this study was to develop a model for necrotizing enterocolitis (NEC) in the neonatal rat using endotoxin and hypoxia, a plausible insult in a neonatal intensive care and to investigate the role of apoptosis as the underlying mechanism. Methods : Newborn rats were given oral endotoxin and intermittent 8% hypoxia$\pm$caspase inhibitor. The intestinal histology was evaluated using hematoxylin-eosin staining. Apoptosis was analyzed with TUNEL staining and by measuring the caspase 3 activity in the intestinal lysates. IEC-6 cells were assessed for apoptosis and the expression of Bax, Bcl-2, Fas and FasL was measured after treatment with endotoxin and hypoxia. Results : Oral endotoxin (5 mg/kg) and exposure to 8% hypoxia of 60-min duration twice induced human NEC-like lesions in the rat intestine. Intestinal tissue revealed increased apoptosis and caspase-3 activity. After caspase inhibitor treatment, the grades of both apoptosis and NEC were significantly reduced. IEC-6 cells exhibited increased apoptosis and caspase 3 activity after endotoxin and hypoxia treatment and significantly increased Bax/Bcl- 2 ratio compared to control cells. Conclusion : This neonatal rat model of NEC which was induced by oral endotoxin and intermittent hypoxia showed increased apoptosis of intestinal epithelial cells that was mediated by caspase 3 activation. Our model has a advantage in the study of NEC because the use of much more clinically plausible insults may provide a suitable model for the investigation of its pathophysiology and therapeutic trials.

A Study on the Characteristics of Oil-water Separation in Non-point Source Control Facility by Coalescence Mechanism of Spiral Buoyant Media (나선형 부유 고분자 여재의 Coalescence 특성을 이용한 비점오염원 저감시설의 유수분리특성 연구)

  • Kang, Sung-Won;Kim, Seog-Ku;Kim, Young-Im;Yun, Sang-Leen;Kim, Soo-Hae;Kim, Mee-Kyung
    • Journal of Korean Society of Environmental Engineers
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    • v.29 no.8
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    • pp.950-955
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    • 2007
  • Non-point source control system which had been designed only for oil-water separation in the fields of oil refinery and garage was upgraded in this research for the removal of runoff pollutants in impervious urban area. Pollutants including oil from driveway and bridge were eliminated by two types of pathway in the system. One is the coalescence mechanism that the oil droplets in the runoff come into contact with each other in the spiral buoyant media surface and form larger coalesced droplets of oil that are carried upstream to the oil layer. The other is the precipitation that solids in runoff were settled by gravity in the system. In this research, coalescing characteristics of oil and water separation were investigated through image analyses, and efficiencies of the non-point source control system were evaluated using dust in driveway and waste engine oil. Media made of high density and high molecular weight polyethylene was indeterminate helical shape and had sleek surface by analysing SEM photographs and BET. Surface area and specific gravity of media which were measured directly were 1,428 $mm^2$ and 45.3 $kg/m^3$ respectively. From the image analyses of the oil droplets photographs which were taken by using microscope, it was proved clearly that the coalescence was the main pathway in the removal of oil from the runoff. Finally, the performances of the non-point source control system filled up with the media were suspended solid $86.6\sim95.2%$, $COD_{Cr}$, $87.3\sim95.4%$, n-Hexane extractable materials $71.8\sim94.8%$ respectively.

Effects of Supplementation of β-Mannanase in Corn-soybean Meal Diets on Performance and Nutrient Digestibility in Growing Pigs

  • Lv, J.N.;Chen, Y.Q.;Guo, X.J.;Piao, X.S.;Cao, Y.H.;Dong, B.
    • Asian-Australasian Journal of Animal Sciences
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    • v.26 no.4
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    • pp.579-587
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    • 2013
  • A total of 288 crossbred (Duroc${\times}$Landrace${\times}$Yorkshire) growing pigs were used in two experiments to investigate the effects of adding ${\beta}$-mannanase to corn-soybean meal-based diets on pig performance and apparent total tract digestibility (ATTD). Both experiments lasted 28 d and were split into two phases namely 1 to 14 days (phase 1) and 15 to 28 days (phase 2). In Exp. 1,144 pigs weighing $23.60{\pm}1.59$ kg BW were assigned to one of four corn-soybean meal-based diets containing 0, 200, 400 or 600 U/kg ${\beta}$-mannanase. Increasing the level of ${\beta}$-mannanase increased weight gain (quadratic effect; p<0.01) and feed efficiency (linear and quadratic effect; p<0.01) during the second phase and the overall experiment. However, performance was unaffected (p>0.05) by treatment during phase 1. Increasing the amount of ${\beta}$-mannanase in the diet improved (linear and quadratic effect; p<0.05) the ATTD of CP, NDF, ADF, calcium, and phosphorus during both phases. Based on the results of Exp. 1, the optimal supplementation level was determined to be 400 U/kg and this was the level that was applied in Exp. 2. In Exp. 2, 144 pigs weighing $23.50{\pm}1.86$ kg BW were fed diets containing 0 or 400 U/kg of ${\beta}$-mannanase and 3,250 or 3,400 kcal/kg digestible energy (DE) in a $2{\times}2$ factorial design. ${\beta}$-Mannanase supplementation increased (p<0.01) weight gain and feed efficiency while the higher energy content increased (p<0.01) feed intake and feed efficiency during both phases and overall. Increased energy content and ${\beta}$-mannanase supplementation both increased (p<0.05) the ATTD of DM, CP, NDF, ADF, phosphorus, and GE during both phases. There were no significant interactions between energy level and ${\beta}$-mannanase for any performance or digestibility parameter. In conclusion, the ${\beta}$-mannanase used in the present experiment improved the performance of growing pigs fed diets based on corn and soybean. The mechanism through which the improvements were obtained appears to be related to improvements in ATTD.

Development of Continuous Galvanization-compatible Martensitic Steel

  • Gong, Y.F.;Song, T.J.;Kim, Han S.;Kwak, J.H.;De Cooman, B.C.
    • Corrosion Science and Technology
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    • v.11 no.1
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    • pp.1-8
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    • 2012
  • The development of martensitic grades which can be processed in continuous galvanizing lines requires the reduction of the oxides formed on the steel during the hot dip process. This reduction mechanism was investigated in detail by means of High Resolution Transmission Electron Microscopy (HR-TEM) of cross-sectional samples. Annealing of a martensitic steel in a 10% $H_2+N_2$ atmosphere with the dew point of $-35^{\circ}C$ resulted in the formation of a thin $_{C-X}MnO.SiO_{2}$ (x>1) oxide film and amorphous $_{a-X}MnO.SiO_{2}$ oxide particles on the surface. During the hot dip galvanizing in Zn-0.13%Al, the thin $_{C-X}MnO.SiO_{2}$ (x>1) oxide film was reduced by the Al. The $_{a-X}MnO.SiO_{2}$ (x<0.9) and $a-SiO_{2}$ oxides however remained embedded in the Zn coating close to the steel/coating interface. No $Fe_{2}Al_{5-X}Zn_{X}$ inhibition layer formation was observed. During hot dip galvanizing in Zn-0.20%Al, the $_{C-X}MnO.SiO_{2}$ (x>1) oxide film was also reduced and the amorphous $_{a-X}MnO.SiO_{2}$ and $a-SiO_{2}$ particles were embedded in the $Fe_{2}Al_{5-X}Zn_{X}$ inhibition layer formed at the steel/coating interface during hot dipping. The results clearly show that Al in the liquid Zn bath can reduce the crystalline $_{C-X}MnO.SiO_{2}$ (x>1) oxides but not the amorphous $_{a-X}MnO.SiO_{2}$ (x<0.9) and $a-SiO_{2}$ oxides. These oxides remain embedded in the Zn layer or in the inhibition layer, making it possible to apply a Zn or Zn-alloy coating on martensitic steel by hot dipping. The hot dipping process was also found to deteriorate the mechanical properties, independently of the Zn bath composition.

An Efficient Dynamic Workload Balancing Strategy (리트윗 행위의 동기, 이유와 가치: 요인 분석)

  • Kim, Hyo D.
    • Journal of the Korea Society of Computer and Information
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    • v.19 no.11
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    • pp.137-147
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    • 2014
  • The study aims at exploring motivation, rationale, and values in twitter users' retweet behavior. It proposes that diffusion of message is based on the complex interactional relationships among attributes of original message, user's rationale, and values. Based on a pilot study, we constructed a total of 34 questions asking message attributes, motivation, and values of retweeting. Then, twitter users participated in an online survey, in which they evaluate their own 5 retweet messages based on the constructed questions(5 messages ${\times}$ 34 questions = 170). Then, a factor analysis is done in order to see the dimensions of the concepts in retweet behavior; and understand how message attributes, motivations, and values are inter-related with each other. The main factors extracted were: (1) public fairness, (2) fun and playfulness, (3) communal help, (4) news and information, etc. Factor 2 and 4 show the traditional journalism characteristics; while factor 1 and 3 do alternative journalistic values. The latter may work as a rectifying factors for traditional journalism; however, backfiring mechanism for group polarization. In addition, (1) users' internal identities, (2) communal unity and (3) belongness were identified as rationales and values for retweet behavior.

Inhibition of Vinyl Carbamate Epoxide- and 2`-(4-Nitrophenoxy)oxirane-induced Mutagenicity by Various Nucleophilic Compounds and Detoxifying Enzymes (Vinyl Carbamate Epoxide와 2`-(4-Nitrophenoxy)oxirane으로 유발된 돌연변이에 대한 친핵성 물질 및 해독작용 효소에 의한 억제)

  • 박광균;이자현;김혜원;김종우;김윤수
    • Environmental Mutagens and Carcinogens
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    • v.17 no.2
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    • pp.97-108
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    • 1997
  • The drugs or xenobiotics introduced to the body, are detoxified through the process of biotransformation in the body. In this process, most of the insoluble compounds become more polar, soluble and easily excretable. But, parts of introduced materials are metabolized to highly reactive electrophilic carcinogens through activation pathways. These metabolites are toxic and can react with DNA, RNA and proteins which are nucleophilic compounds. The objective of this study is to illustrate the aleactivation pathways of two highly reactive epoxide compounds, vinyl carbamate epoxide (VCO) and 2'-(4-nitrophenoxy)oxirane (NPO). They are the ultimate electrophilic carcinogens of ethyl carbamate(urethane) and 4-nitrophenyl vinyl ether, respectively. In this research, we studied the inhibition of the mutagenic activities of VCO or NPO by nuchieophiles [glutahione(GSH) and N-acetylcysteine(NAC)], detoxifying enzymes[epoxide hydrolase and glutathione-S-transferase(GST)] and intracellular organelles (microsomes and cytosol). In addition we also tested the suppression of DNA adducts formation by GSH and NAC. The results are summerized as follow. 1. The microsomes and cytosol which contain epoxide hydrolase and GST, respectively, decreased the mutagenicity of VCO (74% and 95%, respecfivel), and NPO (35% and 93%, respectively). The nucleophilic GSH and NAC decreased the mutagenicity by 86% (VCO) and 80% (NPO), 76% (VCO) and 40% (NPO), respectively. 2. The purified epoxide hydrolase decreased the mutagenicity of two epoxides in a dose-dependent manner, and GSH also decreased the mutagenicity in the presence of GST. 3. Formation of two DNA adducts, 7-(2'-oxoethyi)guanine (OEG) and N2,3-ethenoguanine(EG), were compared in the presence of calf thymus DNA and epoxide (VCO or NPO) in vitro system. The amounts of DNA adducts were decreased in the presence of GSH (25% and 29% in VCO, 32% and 29% in NPO), and NAC (14% and 16% in VCO, 21% and 11% in NPO), respectively. From these results, it is concluded that the ultimate carcinogenic metabolites, VCO and NPO, can be made in the body, but much of them may be inactivated and detoxified by the nucleophilic GSH, NAC and detoxifying enzymes (epoxide hydrolase and GST). Therefore, by these mechanism, the formation of DNA adducts and mutagenic activities of these two epoxides may be lowered in vivo.

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Effect of Inhibitor of Nitric Oxide Synthesis on the Ischemic Reconditioning in Isolated Heart of Rat. (NO 억제제가 허혈전처치의 심장 보호효과에 미치는 영향)

  • 유호진;조은용
    • Journal of Chest Surgery
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    • v.29 no.8
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    • pp.807-815
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    • 1996
  • The protective effect of'ischemic preconditioning'on ischemid-reperfusion injury of heart has been reported in various animal species. but without known mechAnism in detail, In An attempt to investigate the cardioprotective mechanism of ischemic preconditioning, we examined the effects of nitric oxide(UO) synthesis in preconditioned heart of rat The isolated hearts perfused by Langendorfr's method were ex- posed to 30min global ischemia followed by 30min reperfusion with oxygenated Krebs-Henseleit(K-H) sol- ution. Ischemic preconditioning was performed with three episodes of Sm n ischemia and Smin repeyfusion before the induction of prolong ischemia(30min)-reperfusion(30min). Ischemic preconditioning prevented the depression of cardiac function(left ventricular pressure .K heart rate) observed in the ischemia- reperfusion hearts and reduced the release of lactate dehydrogenase during the reperfusion period. On electromicroscopic pictures, myocardial ultrastructures wore relatively well preserved in isthemic preconditioned hearts. N6_nitro-L-arginine methyl ester(L-NAME) an inhibitor of L-arginine citric oxide pathway, was infused at a rate O.Smllmin In a dose of 10mg kg-1 before the initial ischemic preconditioning. neither the protection of cardiac function nor the reduction of LDH releAse in ischemic preconditioning hearts was altered in the presence of added L-NAME On ultrastructural finding, the preservation of morphology in ischemic preconditioning heart was not change by the pretreatment of L-UAME. The failure of the WO synthesis inhibitor to reduce t e effect of ischemic preconditioning may be related to be species specific in that NO may allot be the trigger for ischemic preconditioning in rats.

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