어류 병원세균, Listonella anguillarum에 대한 Pseudomonas aeruginosa MB I-3의 항균 효과

이수정,윤이나,김진도,이정식,김은희,Lee, Su-Jung,Youn, I Na,Kim, Jin-Do,Lee, Jung Sick,Kim, Eunheui 한국어병학회 2014 한국어병학회지 Vol.27 No.1

질병 치료를 위하여 사용되는 항균제는 효과적이고 사용이 편리한 장점이 있지만, 내성균 발생이나 항균물질 잔류와 같은 문제점들을 안고 있다. 따라서 어류 질병 치료 및 예방을 위해 어류와 인체에 안전한 생약제 개발과 함께 어류 질병 원인균의 과다 발생을 억제하며 어체의 건강을 유지시키기 위하여 유용 균주를 이용하고자 하는 연구가 다양하게 진행되고 있다. 본 연구는 이전 연구에서 분리되어 Listonella anguillarum에 대하여 생장 억제효과가 있는 것으로 밝혀진 Pseudomonas aeruginosa MB I-3 (MB I-3)를 이용하여 넙치의 비브리오병에 대한 생물학적 방제효과를 검토하였다. Double layered plate assay와 co-culture을 통하여 MB I-3의 L. anguillarum에 대한 생장 억제능력을 조사하였고, MB I-3 균주 배양액을 ethyl acetate로 추출하여 disk 확산법으로 추출물의 항균 효과를 확인하였다. 액체 및 고체 배양에서 생장이 억제된 L. anguillarum을 전자현미경으로 관찰하였으며, 넙치 치어 사육 수조에 L. anguillarum과 MB I-3 균주를 동시에 첨가하여 폐사율을 비교하였다. MB I-3는 8종의 병원성 비브리오균에 대하여 항균력을 나타내었으며, 96시간 동안 실시한 co-culture에서 L. anguillarum은 배양 후 9시간까지 생장 증가를 보였으나, 그 후 감소하는 경향을 보였다. 한편 MB I-3 배양액 추출물 또한 L. anguillarum에 항균활성을 보여 항균 물질이 ethyl acetate로 추출됨을 알 수 있었다. 전자현미경 관찰에서 L. anguillarum은 세포질의 밀도 감소 및 세포막의 swelling에 의한 세포 용해 현상을 보였다. 한편 MB I-3를 L. anguillarum과 함께 투여한 넙치 치어는 대조군에 비해 누적 폐사율이 약 20% 감소되는 결과를 보였으므로, MB I-3를 이용한 수산용 probiotics 개발 가능성을 시사하였다. To study the possible use of probiotics in fish farming, The in vitro and in vivo antibacterial effects of Pseudomonas aeruginosa MB I-3 (MB I-3) against the fish pathogenic bacterium Listonella anguillarum were evaluated. The inhibitory effects of MB I-3 against vibrios were investigated by the double layer method and the co-culture. The results showed that MB I-3 inhibited the growth of pathogenic vibrios including Listonella anguillarum, Vibrio alginolyticus, Vibrio cholerae, Vibrio fluvialis, Vibrio furnissii, Vibrio harveyi, Vibrio parahaemolyticus and Vibrio vulnificus. Extracellular substances obtained from the cultural supernatant of MB I-3 by ethyl acetate extraction showed inhibitory effects on L. anguillarum. The antibacterial substance of MB I-3 was evaluated to destroy the cell membrane of L. anguillarum in electron micrographs. The probiotic effects of MB I-3 was tested by exposing olive flounder (Paralichthys olivaceus) fry to L. anguillarum with or without MB I-3. The cumulative mortality of olive flounder fry infected with L. anguillarum was 24% in the group with MB I-3, while it was 46% in the control group without MB I-3. These results indicate that MB I-3 has potential applications as a probiotic for the control of fish pathogenic vibrios in fish rearing system.

Manganese-superoxide dismutase (MnSOD), a role player in seahorse (<i>Hippocampus abdominalis</i>) antioxidant defense system and adaptive immune system

Perera, N.C.N.,Godahewa, G.I.,Lee, Seongdo,Kim, Myoung-Jin,Hwang, Jee Youn,Kwon, Mun Gyeong,Hwang, Seong Don,Lee, Jehee Elsevier 2017 Fish & shellfish immunology Vol.68 No.-

<P><B>Abstract</B></P> <P>Manganese superoxide dismutase (MnSOD) is a metaloenzyme that catalyzes dismutation of the hazardous superoxide radicals into less hazardous H<SUB>2</SUB>O<SUB>2</SUB> and H<SUB>2</SUB>O. Here, we identified a homolog of MnSOD from big belly seahorse (<I>Hippocampus abdominalis</I>; <I>HaMnSOD</I>) and characterized its structural and functional features. HaMnSOD transcript possessed an open reading frame (ORF) of 672 bp which codes for a peptide of 223 amino acids. Pairwise alignment showed that HaMnSOD shared highest identity with rock bream MnSOD. Results of the phylogenetic analysis of HaMnSOD revealed a close proximity with rock bream MnSOD which was consistent with the result of homology alignment. The intense expression of <I>HaMnSOD</I> was observed in the ovary, followed by the heart and the brain. Further, immune related responses of <I>HaMnSOD</I> towards pathogenic stimulation were observed through bacterial and viral challenges. Highest <I>HaMnSOD</I> expression in response to stimulants <I>Edwardsiella tarda</I>, <I>Streptococcus iniae</I>, lipopolysaccharide (LPS), and polyinosinic-polycytidylic acid (Poly I:C) was observed in the late stage in the blood tissue. Xanthine/xanthine oxidase assay (XOD assay) indicated the ROS-scavenging ability of purified recombinant HaMnSOD (rHaMnSOD). The optimum conditions for the SOD activity of rHaMnSOD were pH 9 and the 25 °C. Collectively, the results obtained through the expressional analysis profiles and the functional assays provide insights into potential immune related and antioxidant roles of <I>HaMnSOD</I> in the big belly seahorse.</P> <P><B>Highlights</B></P> <P> <UL> <LI> MnSOD was identified from big belly seahorse (HaMnSOD). </LI> <LI> HaMnSOD was cloned and expressed to evaluate its distinct functional features. </LI> <LI> XOD (Xanthine oxidase) assay confirmed the superoxide scavenging ability of HaMnSOD. </LI> <LI> Transcriptional level of <I>HaMnSOD</I> was modulated by pathological stress. </LI> </UL> </P>

Radiosynthesis and <i>in vitro</i> evaluation of 1‐(tetrahydro‐5‐hydroxy‐6‐(hydroxymethyl)‐2H‐pyran‐3‐yl)‐5‐[¹²⁵I]iodouracil: A new potential agent for HSV1‐tk

Jo, Nam Hyun,Kim, Jung Young,El‐,Gamal, Mohammed I.,Choi, Won‐,Kyoung,Park, Jin‐,Hun,Kim, Eun Jung,Cho, Jung‐,Hyuck,Ha, Hyun‐,Joon,Choi, Tae Hyun,Oh, Chang‐,Hyun John Wiley Sons, Ltd. 2011 Journal of labelled compounds & radiopharmaceutica Vol.54 No.2

<P><B>Abstract</B></P><P>Synthesis, radiolabelling, and <I>in vitro</I> evaluation of a new <SUP>125</SUP>I‐labelled iodouracil hexitol nucleoside analogue are reported. The target compound was successfully synthesized by an iodination–destannylation method and then purified by reverse phase HPLC. The radiochemical purity of the product was >99% with decay‐corrected yields of 48±3%. <I>In vitro</I> cellular uptake testing was carried out using MCA and MCA‐tk cell lines for comparison of compound 1 with [<SUP>18</SUP>F]FHBG. The newly synthesized compound 1 showed higher accumulation in herpex simplex virus type 1 thymidine kinase (HSV1‐tk) gene expression cell line (MCA‐tk cell line) than in the wild type MCA cell line compared with [<SUP>18</SUP>F]FHBG. The MCA‐tk to MCA cellular uptake ratio for compound 1 was higher than that of [<SUP>18</SUP>F]FHBG from 2 h after incubation. The radioiodine‐labelled compound 1 (I‐125, <I>t</I><SUB>1/2</SUB>=59.37 days) has a longer physical half‐life than F‐18‐(<I>t</I><SUB>1/2</SUB>=110 min) labelled FHBG. Radioiodine‐labelled compound 1 could be used for monitoring gene expression for a long time. The selectivity for MCA‐tk cell line makes compound 1 a promising imaging agent for HSV1‐tk expression. Copyright © 2010 John Wiley & Sons, Ltd.</P>

Identification and characterization of a carboxypeptidase N1 from red lip mullet (<i>Liza haematocheila</i>); revealing its immune relevance

Perera, N.C.N.,Godahewa, G.I.,Jung, Sumi,Kim, Myoung-Jin,Nam, Bo-Hye,Lee, Jehee ACADEMIC PRESS LTD 2019 FISH AND SHELLFISH IMMUNOLOGY Vol.84 No.-

<P><B>Abstract</B></P> <P>Complement system orchestrates the innate and adaptive immunity <I>via</I> the activation, recruitment, and regulation of immune molecules to destroy pathogens. However, regulation of the complement is essential to avoid injuries to the autologous tissues. The present study unveils the characteristic features of an important complement component, anaphylatoxin inactivator from red lip mullet at its molecular and functional level. Mullet carboxypeptidase N1 (MuCPN1) cDNA sequence possessed an open reading frame of 1347 bp, which encoded a protein of 449 amino acids with a predicted molecular weight of 51 kDa. <I>In silico</I> analysis discovered two domains of PM14-Zn carboxypeptidase and a C-terminal domain of M14 N/E carboxypeptidase, two zinc-binding signature motifs, and an N-glycosylation site in the MuCPN1 sequence. Homology analysis revealed that most of the residues in the sequence are conserved among the other selected homologs. Phylogeny analysis showed that MuCPN1 closely cladded with the <I>Maylandia zebra</I> CPN1 and clustered together with the teleostean counterparts. A challenge experiment showed modulated expression of MuCPN1 upon polyinosinic:polycytidylic acid and <I>Lactococcus garviae</I> in head kidney, spleen, gill, and liver tissues. The highest upregulation of MuCPN1 was observed 24 h post infection against poly I:C in each tissue. Moreover, the highest relative expressions upon <I>L. garviae</I> challenge were observed at 24 h post infection in head kidney tissue and 48 h post infection in spleen, gill, and liver tissues. MuCPN1 transfected cells triggered a 2.2-fold increase of nitric oxide (NO) production upon LPS stimulation compared to the un-transfected controls suggesting that MuCPN1 is an active protease which releases arginine from complement C3a, C4a, and C5a. These results have driven certain way towards enhancing the understanding of immune role of MuCPN1 in the complement defense mechanism of red lip mullet.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Carboxypeptidase N1 complement component was identified from the red lip mullet. </LI> <LI> Ubiquitous expression of MuCPN1 was observed in healthy mullet tissues. </LI> <LI> Modulated transcriptions of MuCPN1 revealed the importance in the immune responses. </LI> <LI> MuCPN1 was enhanced the nitric oxide production at an inflammatory condition. </LI> </UL> </P>

I-V 장치 특성 및 I-V 장치를 이용한 ZnO 나노막대의 전기적 특성 연구

김정란 ( Zhen Lan Jin ),박창인 ( C. I. Park ),김병혁 ( B. H. Kim ),한상욱 ( S. W. Han ) 전북대학교 과학교육연구소 2010 과학과 과학교육 논문지 Vol.35 No.-

수직으로 배열이 잘 된 ZnO 나노막대는 metal organic chemical vapor deposition방법을 이용하여 Al2O3(0001)기판 위에 생장시킨 후 질소이온을 주입하여 400 ℃에 열처리하였다. ZnO 나노막대의 전기적 특성은 I-V 장비를 이용하여 측정하였다. I-V 장비는 매우 미세한 전류를 측정할 수 있는 장비로서 주위 환경의 안정성 그리고 측정 시 두 팁사이의 거리, 팁이 샘플에 주는 압력, 인가한 전압의 크기 및 팁의 지름의 크기와 매우 밀접한 관계가 있음을 볼 수 있었다. ZnO 나노막대의 전기적 특성을 측정한 결과 ohmic contact 현상이 발생하였음을 알 수 있었다. We have synthesized vertically aligned ZnO nanorods by metal organic chemical vapor deposition on Al2O3(0001) substrate. We implanted N+ to vertically aligned ZnO nanorods and annealed at different temperatures. The electrical property of Zno nanorods was measured by using I-V measurements. I-V systems is very sensitive with around condition, between two tips distance, pressure of tip for the sample, source range of electric field and used tip diameter at measurement. We observed that ZnO nanorods showed ohmic contact.

Comparison of single-dose and non-single-dose protocol of methotrexate in treatment of ectopic pregnancy: a systematic review and meta-analysis

( Jin-sung Yuk ),( Won I. Park ),( Jung Hun Lee ),( Hyeong Sik Ahn ),( Hyun Jung Kim ) 대한산부인과학회 2016 대한산부인과학회 학술대회 Vol.102 No.-

목적: The purpose of this study was to compare treatment success rates, side effects rates and time to resolution between single-dose protocol and non-single-dose protocol (multi-dose protocol and two-dose protocol) of methotrexate (MTX) in treatment of ectopic pregnancy (EP) using randomized controlled trials (RCTs). 방법: We searched MEDLINE, EMBASE, SCOPUS, Web of Science, the Cochrane Central Register of Controlled Trials, ClinicalTrials.gov, International Clinical Trials Registry Platform and EU Clinical Trials Register. The relative risk (RR) and 95% confidence intervals (CIs) were used for calculating dichotomous outcomes, while mean difference (MD) and 95% CIs were used for calculating continuous outcomes. 결과: We selected 9 RCTs among total 1521 articles. The success rate of single-dose protocol was similar to that of non-single-dose protocol in treating EP [RR (95% CI): 1.02 (0.98~1.06; 9 trials, 891 patients), I2=0%]. Although non-single-dose protocol tended to have a higher side effects compared to single-dose protocol, there was no significant difference in both group, [RR (95% CI): 0.76 (0.58~1.01; 7 trials, 704 patients), I2=12%]. However, in subgroup analysis about tubal pregnancy, non-single dose protocol appeared to have a higher side effects rate than single-dose protocol [RR (95% CI): 0.69 (0.51~0.93; 3 trials, 369 patients), I2=0%]. Single-dose protocol had longer resolution time than non-single-dose protocol [MD (95% CI): 4.5 (0.40~8.60; 5 trials, 528 patients), I2=83%]. 결론: Single-dose protocol has similar success rates to non-single-dose protocol in treating EP, and tends to lower side effects rate than non-single- dose protocol. Also, in treating tubal pregnancy, single-dose protocol has lower side effects rate than non-single-dose protocol. However, single- dose protocol has longer resolution time than non-single-dose protocol.

Cryptotanshinone, a lipophilic compound of Salvia miltiorrriza root, inhibits TNF-α-induced expression of adhesion molecules in HUVEC and attenuates rat myocardial ischemia/reperfusion injury in vivo

Jin, Y.C.,Kim, C.W.,Kim, Y.M.,Nizamutdinova, I.T.,Ha, Y.M.,Kim, H.J.,Seo, H.G.,Son, K.H.,Jeon, S.J.,Kang, S.S.,Kim, Y.S.,Kam, S.C.,Lee, J.H.,Chang, K.C. North-Holland ; Elsevier Science Ltd 2009 european journal of pharmacology Vol.614 No.1

The aim of the present study was to evaluate the protective effect of cryptotanshinone (CTS), one of active ingredients of Salvia miltiorrhiza root, on myocardial ischemia-reperfusion injury in rat due to inhibition of some inflammatory events that occur by NF-kB-activation during ischemia and reperfusion. Myocardial ischemia and reperfusion injury was induced by occluding the left anterior descending coronary artery for 30 min followed by either 2 h (biochemical analysis) or 24 h (myocardial function and infarct size measurement) reperfusion. CTS injected (i.v.) 10 min before ischemia and reperfusion insult. CTS significantly reduced the infarct size and improved ischemia and reperfusion-induced myocardial contractile dysfunction. Furthermore, CTS inhibited NF-kB translocation, expression of pro-inflammatory cytokines (TNF-α, IL-1β, IL-6), neutrophil infiltration and MPO activity in ischemic myocardial tissues. CTS also significantly reduced plasma levels of TNF-α, IL-1β due to ischemia and reperfusion. Interestingly, H<SUB>2</SUB>O<SUB>2</SUB>-stimulated NF-kB-luciferase activity and TNF-α-induced expression of vascular cell adhesion molecule-1 (VCAM-1) and intracellular adhesion molecule-1 (ICAM-1) expressions in human umbilical vein endothelial cells (HUVEC) were significantly inhibited by CTS. Taken together, it is concluded that CTS may attenuate ischemia and reperfusion-induced microcirculatory disturbances by inhibition of proinflammatory cytokine production, reduction of neutrophil infiltration and possibly inhibition of adhesion molecules through inhibition of NF-kB-activation during ischemia and reperfusion.

Effect of Alkali Metal Ions on Nucleophilic Substitution Reactions of 4-Nitrophenyl X-Substituted Benzoates with Alkali Metal Ethoxides in Anhydrous Ethanol

Jin-A Seo,Song-I Kim,Yeon-Ju Hong,엄익환 대한화학회 2010 Bulletin of the Korean Chemical Society Vol.31 No.2

Pseudo-first-order rate constants (kobsd) have been measured spectrophotometrically for nucleophilic substitution reactions of 4-nitrophenyl benzoate (5a), 4-nitrophenyl 4-methoxybenzoate (5b), and 4-nitrophenyl 4-hydroxybenzoate (5c) with alkali metal ethoxides, EtO–M+ (M+= Li+, Na+ and K+) in anhydrous ethanol (EtOH) at 25.0 ± 0.1 oC. The plots of kobsd vs. [EtO–M+] exhibit upward curvatures in all cases, indicating that M+ ions catalyze the reactions and ionpaired EtO–M+ species are more reactive than dissociated EtO–. Second-order rate constants for reactions with dissociated EtO– and ion-paired EtO–M+ (i.e., kEtO– and kEtO–M+, respectively) have been calculated from ion-pair treatment for the reactions of 5a and 5b. However, such ion-pair treatment has failed to determine kEtO– and kEtO–M+ values for the reactions of 5c. It has been concluded that reactions of 5a and 5b are catalyzed by one metal ion, which increases electrophilicity of the reaction center through coordination on the carbonyl oxygen. In contrast, reactions of 5c have been suggested to involve two metal ions, i.e., the one coordinated on the carbonyl oxygen increases the electrophilicity of the reaction center while the other one associated on the phenoxy oxygen decreases the charge repulsion between the anionic reagents (i.e., EtO– and deprotonated 5c). It has been found that the rate equation derived from the mechanism involving two metal ions fits nicely to the kinetic results obtained for the reactions of 5c.

Derivation of Q* Parameter for Evaluating Creep Crack Growth Rate of Modified 9Cr-1Mo Steel

I.M.W. Ekaputra,Woo-Gon Kim(김우곤),Jae-Young Park(박재영),Seon-Jin Kim(김선진),Min-Hwan Kim(김민환),Yong-Wan Kim(김용완) 대한기계학회 2013 대한기계학회 춘추학술대회 Vol.2013 No.12

The concept of the Q* parameter was first proposed by Yokobori et al. The Q* parameter is defined as the exponent of the exponential function in the thermal activation process equation. In this study, the creep crack growth (CCG) behavior of the modified 9Cr-1Mo steel is evaluated using the Q* parameter. The CCG tests were carried out under various loads at 550oC and 600oC. The K and C* parameters have been used to characterize the CCGR of modified 9Cr-1Mo steel. The results show that the K parameter exhibits the largest scatter data, and there is no systematic trend in each series of tests, while C* showed a narrower scattering of data of CCGR than the K parameter. However, C* decreased during the early stage of crack growth and subsequently increased, i.e., a dual value due to the nose appearance, and it did not distinguish among the series of data under various loads at 550oC and 600oC clearly in a one linear line. The Q* parameter was able to evaluate the CCG by a simple monotical linear function without a dual value owing to nose existing in the early stage, and it exhibited an increase or decrease regardless of the testing conditions. The Q* was regarded as an independent parameter that is not depending on stress and temperature, whereas the C* was regarded as a dependent parameter depending on the creep stress and temperature.

Nucleophilic Substitution Reactions of Y-Substituted-Phenyl Benzoates with Potassium Ethoxide in Anhydrous Ethanol: Reaction Mechanism and Role of K+ Ion

Song-I Kim,Hyo-Jin Cho,엄익환 대한화학회 2014 Bulletin of the Korean Chemical Society Vol.35 No.1

Pseudo-first-order rate constants (k obsd ) have been measured spectrophotometrically for the reactions of Y- substituted-phenyl benzoates (5a-j) with potassium ethoxide (EtOK) in anhydrous ethanol at 25.0 ± 0.1 o C. The plots of k obsd vs. [EtOK] curve upward regardless of the electronic nature of the substituent Y in the leaving group. Dissection of k obsd into the second-order rate constants for the reactions with the dissociated EtO – and ion-paired EtOK (i.e., k EtO- and k EtOK , respectively) has revealed that the ion-paired EtOK is more reactive than the dissociated EtO – . The Brønsted-type plots for the reactions with the dissociated EtO – and ion-paired EtOK exhibit highly scattered points with βlg = –0.5 ± 0.1. The Hammett plots correlated with σo constants result in excellent linear correlations, indicating that no negative charge develops on the O atom of the leaving Y- substituted-phenoxide ion in transition state. Thus, it has been concluded that the reactions with the dissociated EtO – and ion-paired EtOK proceed through a stepwise mechanism, in which departure of the leaving group occurs after the RDS, and that K + ion catalyzes the reactions by increasing the electrophilicity of the reaction center through a four-membered cyclic TS structure.