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      • Diagnostic usefulness of a T cell-based assay for latent tuberculosis infection in kidney transplant candidates before transplantation

        Kim, S.-H.,Lee, S.-O.,Park, I.-A.,Park, S.J.,Choi, S.-H.,Kim, Y.S.,Woo, J.H.,Park, S.-K.,Park, J.S.,Kim, S.C.,Han, D.J. Blackwell Publishing Inc 2010 Transplant infectious disease Vol.12 No.2

        <P>S.-H. Kim, S.-O. Lee, I.-A. Park, S.J. Park, S.-H. Choi, Y.S. Kim, J.H. Woo, S.-K. Park, J.S. Park, S.C. Kim, D.J. Han. Diagnostic usefulness of a T cell-based assay for latent tuberculosis infection in kidney transplant candidates before transplantation.Transpl Infect Dis 2010: <B>12:</B> 113–119. All rights reserved</P><P>Background</P><P>The presence of latent tuberculosis (TB) infection (LTBI) should be evaluated before kidney transplantation. Although a new T cell-based assay for diagnosing LTBI gave promising results, this assay has not yet been compared with the tuberculin skin test (TST) for diagnosing LTBI in renal transplant candidates before transplantation.</P><P>Patients and methods</P><P>All adult patients admitted to a single institute for renal transplantation over a 1-year period were prospectively enrolled. A clinically predictive risk of LTBI was defined as: (i) recent close contact with a person with pulmonary TB; (ii) abnormal chest radiography; (iii) a history of untreated or inadequately treated TB; or (iv) a new infection (i.e., a recent conversion of TST).</P><P>Results</P><P>Of 209 renal recipients, 47 (22%) had a positive TST≥5 mm, 21 (10%) had a positive TST≥10 mm, 65 (30%) had a positive T-SPOT.<I>TB</I> test, and 25 (12%) had an indeterminate T-SPOT.<I>TB</I> test. The induration size of TST was significantly associated with a high positivity rate on T-SPOT.<I>TB</I> (<I>P</I><0.001). Agreement between T-SPOT.<I>TB</I> test and TST≥10 mm was fair (<I>k</I>=0.24, 95% confidence interval 0.11–0.36). However, neither univariate nor multivariate analysis showed any association between the clinical risk for LTBI and positivity on T-SPOT.<I>TB</I> or TST.</P><P>Conclusion</P><P>T-SPOT.<I>TB</I> test was more frequently positive than TST in renal transplant candidates. However, further longitudinal studies are awaited to determine whether the ability of T-SPOT.<I>TB</I> assay to detect LTBI in renal transplant recipients can better predict the development of TB than can TST after transplantation.</P>

      • <i>CYP2A6</i> and <i>ERCC1</i> polymorphisms correlate with efficacy of S-1 plus cisplatin in metastatic gastric cancer patients

        Park, S R,Kong, S-Y,Nam, B-H,Choi, I J,Kim, C G,Lee, J Y,Cho, S J,Kim, Y W,Ryu, K W,Lee, J H,Rhee, J,Park, Y-I,Kim, N K Nature Publishing Group 2011 The British journal of cancer Vol.104 No.7

        <P><B>Background:</B></P><P>We evaluated the association between polymorphisms of cytochrome P450 2A6 (<I>CYP2A6</I>)/excision repair cross-complementation group 1 (<I>ERCC1</I>)/X-ray repair cross-complementing group 1(<I>XRCC1</I>) and treatment outcomes of metastatic gastric cancer (MGC) patients treated with S-1/cisplatin.</P><P><B>Methods:</B></P><P>Among MGC patients (<I>n</I>=108), who received S-1 (40 mg m<SUP>−2</SUP> b.i.d., days 1–14) and cisplatin (60 mg m<SUP>−2</SUP>, day 1) every 3 weeks, we analysed the wild-type allele (<I>W</I>) and variants (<I>V</I>) of <I>CYP2A6</I> (<I>*4</I>, <I>*7, *9, *10</I>), and the polymorphisms of <I>ERCC1</I> (rs11615, rs3212986) and <I>XRCC1</I> (rs25487).</P><P><B>Results:</B></P><P>Patients having fewer <I>CYP2A6</I> variants had better response rates (<I>W</I>/<I>W vs W</I>/<I>V</I> other than <I>*1/*4 vs V</I>/<I>V</I> or <I>*1/*4</I>=66.7 <I>vs</I> 58.3 <I>vs</I> 32.3% <I>P</I>=0.008), time to progression (TTP) (7.2 <I>vs</I> 6.1 <I>vs</I> 3.5 months, <I>P</I>=0.021), and overall survival (23.2 <I>vs</I> 15.4 <I>vs</I> 12.0 months, <I>P</I>=0.004). <I>ERCC1 19442C</I>><I>A</I> (rs3212986) was also associated with response rate (<I>C/C</I>, 46.7% <I>vs C/A</I>, 55.3% <I>vs A/A</I>, 87.5%) (<I>P</I>=0.048) and TTP (4.4 <I>vs</I> 7.6 <I>vs</I> 7.9 months) (<I>P</I>=0.012). Patients carrying both risk genotypes of <I>CYP2A6</I> (<I>V</I>/<I>V</I> or <I>1/*4</I>) and <I>ERCC1 19442C</I>><I>A</I> (<I>C/C</I>) <I>vs</I> those carrying none showed an adjusted odds ratio of 0.113 (<I>P</I>=0.004) for response, and adjusted hazard ratios of 3.748 (<I>P</I>=0.0001) for TTP and 2.961 (<I>P</I>=0.006) for death.</P><P><B>Conclusion:</B></P><P>Polymorphisms of <I>CYP2A6</I> and <I>ERCC1 19442C</I>><I>A</I> correlated with the efficacy of S-1/cisplatin.</P>

      • KCI등재

        Role of the Meckel’s Cartilage in Embryonic Mandibular Development of Mice

        J. W Choi,S. B Han,J. H Sung,H. I Shin 대한구강악안면병리학회 2005 대한구강악안면병리학회지 Vol.29 No.5

        Mecke!'s car t ilage is one of the ea rliest structu re to appear in a mandible derived from the lï rst branchi a l a rch and serves as the primorclium I"or the formation 01‘ mandible‘ mall eus. incus. and sphenomandibular li gament However. its direct role a nd the mechanism in mandibular clevelopment a re not well elucidated. 1'0 address t his Issue‘ we observed morphol ogical and histological changes and gene expression patterns in the Mecke!'s cart ilage 01" a cleveloping mouse. I"rom E13.5 to E18.5 embryos. using skeletal preparation samples a ncl routinely prepa red s lide secti ons for light mi croscopic observation in various sectional planes. The following methods were per |‘ ormecl : H&E staining I"or general hi st이 og i cal observation ‘ Von Kossafor detection of minerali zation. TRAP activ ity staining for locali zaLion 01’ osteoclastic cell s. immunohistochemistry for !Iα@-1 a ncl -9 forevaluati on of enzy matic activity 01" osteoclasLic cell s. a ncl in situ hybricli zation for detection of collagen type 1. Il. ancl X mRNA ex presslon‘ respecLively. AL E1 3.5 Mec kel's cartilage appeared as a V-shaped rod fused a t the micl line and thin minera li zed ma ndibular buccal plaLe was I"ormed lateral to. and at some clistance from. Meckel’s carti lage in an intramembranous ossi lïcation mocle. WiLh the progression of tooth development. t he Meckel’s in carti lage adjacent incisors revealecl hyperLrophi c chonclrocyte di ff"er entiation with minerali zation of the chondroid matrix. The Meckel’s car Li lage was replacecl with bone by o~ L eoc l asLs . showing strong immunoreact ivity for MMP- l ancl -9 from E16 5 Wi Lh ti me‘ Lhis bony replacement of Meckel's cartilage in an endochondral ossification mode was ex Lenclecl up Lo the mid-porLion of Lhe molar sockets til l EI8.5. The bony replacement of minera li zed hypertrophic chondrocyte zone expressing X collagen mHNA conLri buted to the formation of thick mandibular lingual plate . 1'hese f"i ndings suggesL LhaL mandibular formalion and development is closely relatecl with not only Mecke!'s carLi lage. buL also wiLh Lhc developing LooLh. and thaL C'erLai n in f"l uence from the developing tooth may play a role in detcrmin in g Lhe faLc of Meckel’s ca rLi lage cluring ma ndi bular development.

      • SCISCIESCOPUS

        Induction of bone formation by <i>Escherichia coli</i>‐expressed recombinant human bone morphogenetic protein‐2 using block‐type macroporous biphasic calcium phosphate in orthotopic and ectopic rat models

        Park, J‐,C.,So, S,S.,Jung, I,H.,Yun, J‐,H.,Choi, S,H.,Cho, K‐,S.,Kim, C‐,S. Blackwell Publishing Ltd 2011 Journal of periodontal research Vol.46 No.6

        <P><I>Park J‐C, So S‐S, Jung I‐H, Yun J‐H, Choi S‐H, Cho K‐S, Kim C‐S. Induction of bone formation by</I> Escherichia coli<I>‐expressed recombinant human bone morphogenetic protein‐2 using block‐type macroporous biphasic calcium phosphate in orthotopic and ectopic rat models. J Periodont Res 2011; 46: 682–690. © 2011 John Wiley & Sons A/S</I></P><P><B>Background and Objective: </B> The potential of the <I>Escherichia coli</I>‐expressed recombinant human bone morphogenetic protein‐2 (ErhBMP‐2) to support new bone formation/maturation using a block‐type of macroporous biphasic calcium phosphate (bMBCP) carrier was evaluated in an orthotopic and ectopic rat model.</P><P><B>Material and Methods: </B> Critical‐size (Φ 8 mm) calvarial defects and subcutaneous pockets in 32 Sprague–Dawley rats received implants of rhBMP‐2 (2.5 μg) in a bMBCP carrier or bMBCP alone (control). Implant sites were evaluated using histological and histometric analysis following 2‐ and 8‐wk healing intervals (eight animals/group/interval).</P><P><B>Results: </B> ErhBMP‐2/bMBCP supported significantly greater bone formation at 2 and 8 wk (10.8% and 25.4%, respectively) than the control at 2 and 8 wk (5.3% and 14.0%, respectively) in calvarial defects (<I>p</I> < 0.01). Bone formation was only observed for the ErhBMP‐2/bMBCP ectopic sites and was significantly greater at 8 wk (7.5%) than at 2 wk (4.5%) (<I>p</I> < 0.01). Appositional and endochondral bone formation was usually associated with a significant increase in fatty marrow at 8 wk. The bMBCP carrier showed no evidence of bioresorption.</P><P><B>Conclusion: </B> ErhBMP‐2/bMBCP induced significant bone formation in both calvarial and ectopic sites. Further study appears to be required to evaluate the relevance of the bMBCP carrier.</P>

      • SCISCIESCOPUS

        Enhanced adipogenic differentiation and reduced collagen synthesis induced by human periodontal ligament stem cells might underlie the negative effect of recombinant human bone morphogenetic protein‐2 on periodontal regeneration

        Song, D‐,S.,Park, J‐,C.,Jung, I,H.,Choi, S,H.,Cho, K‐,S.,Kim, C‐,K.,Kim, C‐,S. Blackwell Publishing Ltd 2011 Journal of periodontal research Vol.46 No.2

        <P> <I>Song D‐S, Park J‐C, Jung I‐H, Choi S‐H, Cho K‐S, Kim C‐K, Kim C‐S. Enhanced adipogenic differentiation and reduced collagen synthesis induced by human periodontal ligament stem cells might underlie the negative effect of recombinant human bone morphogenetic protein‐2 on periodontal regeneration. J Periodont Res 2011; 46: 193–203. © 2010 John Wiley & Sons A/S</I> </P><P><B>Background and Objective: </B> Recombinant human bone morphogenetic protein‐2 (rhBMP‐2) is a potent inducer for the regeneration of mineralized tissue, but has a limited effect on the regeneration of cementum and periodontal ligament (PDL). The aim of the present study was to determine the effects of rhBMP‐2 on the <I>in vitro</I> and <I>in vivo</I> biologic activity of well‐characterized human PDL stem cells (hPDLSCs) and to elucidate the underlying mechanism of minimal periodontal regeneration by rhBMP‐2.</P><P><B>Material and Methods: </B> hPDLSCs were isolated and cultured, and then transplanted into an ectopic subcutaneous mouse model using a carrier treated either with or without rhBMP‐2. Comprehensive histologic, histometric and immunohistochemical analyses were performed after an 8‐wk healing period. The effects of rhBMP‐2 on the adipogenic and osteogenic/cementogenic differentiation of hPDLSCs were also evaluated. The effect of rhBMP‐2 on both soluble and insoluble collagen synthesis was analyzed, and the expression of mRNA and protein for collagen types I, II, III and V was assessed.</P><P><B>Results: </B> In the present study, rhBMP‐2 promoted both adipogenic and osteogenic/cementogenic differentiation of hPDLSCs <I>in vitro</I>, and the <I>in vivo</I> potential of hPDLSCs to form mineralized cementum and organized PDL tissue was down‐regulated following treatment with rhBMP‐2. Collagen synthesis, which plays a crucial role in the regeneration of cementum and the periodontal attachment, was significantly reduced, with associated modification of the relevant mRNA and protein expression profiles.</P><P><B>Conclusion: </B> In summary, the findings of the present study suggest that enhanced adipogenic differentiation and inhibition of collagen synthesis by hPDLSCs appear to be partly responsible for the minimal effect of rhBMP‐2 on cementum and PDL tissue regeneration by hPDLSCs.</P>

      • Phase I/II study of S-1 combined with weekly docetaxel in patients with metastatic gastric carcinoma

        Park, S R,Kim, H K,Kim, C G,Choi, I J,Lee, J S,Lee, J H,Ryu, K W,Kim, Y-W,Bae, J-M,Kim, N K Cancer Research UK 2008 The British journal of cancer Vol.98 No.8

        We designed a phase I/II trial of S-1 combined with weekly docetaxel to determine the maximum tolerated dose (MTD) and recommended dose (RD) and to evaluate the efficacy and toxicity in metastatic gastric carcinoma (MGC). Patients with measurable disease received S-1 orally b.i.d. on days 1–14 and docetaxel intravenously on days 1 and 8 every 3 weeks. In phase I (n=30), each cohort received escalating doses of S-1 (30–45 mg m<SUP>−2</SUP> b.i.d.) and docetaxel (25–40 mg m<SUP>−2</SUP>); MTD was 45 mg m<SUP>−2</SUP> b.i.d. S-1/35 mg m<SUP>−2</SUP> docetaxel and RD was 40 mg m<SUP>−2</SUP> b.i.d. S-1/35 mg m<SUP>−2</SUP> docetaxel. Dose-limiting toxicities included grade 3 elevated liver enzymes, gastric perforation, grade 3 diarrhoea/fatigue, febrile neutropenia with grade 3 anorexia/fatigue, and neutropenic infection with grade 3 stomatitis/anorexia. In phase II (n=52), the overall response rate was 66.7% (95% confidence interval (CI): 53.8–79.6%) and the median time to progression and overall survival were 6.5 months (95% CI: 4.9–8.1) and 13.7 months (95% CI: 9.9–17.5), respectively. The most common grade 3/4 toxicity was neutropenia (29.4%), and febrile neutropenia/neutropenic infection occurred in 19.6% of patients. Non-haematological toxicities were generally mild. There was one treatment-related death due to pneumonitis. S-1 combined with weekly docetaxel is active in MGC with moderate toxicities.British Journal of Cancer (2008) 98, 1305–1311. doi:10.1038/sj.bjc.6604312 www.bjcancer.com Published online 25 March 2008

      • Differentiation of Recurrent Tumor and Posttreatment Changes in Head and Neck Squamous Cell Carcinoma: Application of High b-Value Diffusion-Weighted Imaging

        Hwang, I.,Choi, S.H.,Kim, Y.-J.,Kim, K.G.,Lee, A.L.,Yun, T.J.,Kim, J.-h.,Sohn, C.-H. American Society of Neuroradiology 2013 AJNR, American journal of neuroradiology Vol.34 No.12

        <P><B>BACKGROUND AND PURPOSE:</B></P><P>High b-value DWI has been expected to have an additional diagnostic role and demonstrated some promising results in head and neck cancer. The aim of this study was to evaluate the diagnostic performance of DWI at a high b-value (<I>b</I>=2000 s/mm<SUP>2</SUP>) compared with a standard b-value (<I>b</I>=1000 s/mm<SUP>2</SUP>) and the ratio of ADC values of high and standard b-values for their ability to differentiate between recurrent tumor and posttreatment changes after the treatment of head and neck squamous cell carcinoma.</P><P><B>MATERIALS AND METHODS:</B></P><P>A total of 33 patients diagnosed with head and neck squamous cell carcinoma were enrolled in the present study; all had contrast-enhancing lesions on follow-up MR imaging. All patients underwent single-shot echo-planar DWI at <I>b</I>=1000 s/mm<SUP>2</SUP> and <I>b</I>=2000 s/mm<SUP>2</SUP>, and corresponding ADC maps were generated (ADC<SUB>1000</SUB> and ADC<SUB>2000</SUB>, respectively). The mean ADC<SUB>1000</SUB>, ADC<SUB>2000</SUB>, and ADC<SUB>ratio</SUB> (ADC<SUB>ratio</SUB> = ADC<SUB>2000</SUB>/ADC<SUB>1000</SUB> × 100) values were evaluated within a manually placed ROI with contrast-enhanced T1-weighted images as references. For the statistical analysis, we performed a Student <I>t</I> test and multivariate logistic regression.</P><P><B>RESULTS:</B></P><P>The mean ADC<SUB>1000</SUB> in recurrent tumor was significantly lower than that in posttreatment changes (<I>P</I> < .001), whereas the mean ADC<SUB>2000</SUB> resulted in no significant difference (<I>P</I> = .365). The mean ADC<SUB>ratio</SUB> was significantly higher in recurrent tumor than that in posttreatment changes (73.5 ± 7.2% vs 56.9 ± 8.8%, respectively; <I>P</I> < .001). Multivariate logistic regression analysis revealed that the ADC<SUB>ratio</SUB> was the only independently differentiating variable (<I>P</I> = .024). The sensitivity, specificity, and accuracy of ADC<SUB>ratio</SUB> were 95.0%, 69.2%, and 84.8%, respectively, by use of the optimal cutoff value of 62.6%.</P><P><B>CONCLUSIONS:</B></P><P>We suggest that the ADC<SUB>ratio</SUB> calculated from the ADC<SUB>1000</SUB> and ADC<SUB>2000</SUB> is a promising value for the differentiation of recurrent tumor and posttreatment changes in head and neck squamous cell carcinoma.</P>

      • KCI등재

        벼 육묘트레이를 이용한 어린 잎 채소 생육에 미치는 광 조절 영향

        최기영 ( K. Y. Choi ),김시홍 ( S. H. Kim ),김재경 ( J. K. Kim ),유형주 ( H. J. Yoo ),김일섭 ( I. S. Kim ) 강원대학교 농업생명과학연구원(구 농업과학연구소) 2016 강원 농업생명환경연구 Vol.28 No.2

        벼 육묘 트레이를 활용한 어린잎 채소 재배 가능성을 탐색하고자 재배 기간 중 차광 처리하여 다채, 청경채, 알파인배추, 청치마 상추 등 4종을 17일간 재배하였다. 차광 처리는 무차광 17일(NS), 차광 8일+무차광 9일 (S8NS9), 무차광 8일+차광 9일(NS8S9), 차광 17일(S17) 하였다. 처리에 따른 누적 광량은 NS>S8NS9>NS8S9>S17처리구 순으로 높았으나, 누적 온도는 처리에 따른 차이가 크지 않았다 차광 8일 째 어린잎채소 4종의 생육과 엽록소함량은 감소하였다. 광 조절 17일 째 어린잎채소 4종의 초장, 엽장, 엽폭, 경경, 생체중, 엽록소 함량 등의 생육은 무차광 NS 처리구에서 가장 높았다. 생육 초기 광조절에 따른 생육 후기 초장과 생체중 증가율이 어린잎채소 4종 모두 NS 또는 S8NS9 처리구에서 높았다. 따라서 벼 육묘장 다층 시스템에서의 공간 활용을 고려한 어린잎채소 재배시 차광이 많은 하부 공간은 생육초기에 활용하고, 수확기에는 광 확보가 이루어지는 상부에서 재배가 이루어짐이 적합하리라 본다. This study was conducted to investigate the effect of light control on growth of baby leaf vegetables (tatsoi, pak-choi, chinese cabbage ‘alpine’ and leaf lettuce ‘cheongchima’) in rice seedling tray. Light treatment controlled in non-shading (NS), shading for 8 days+non-shading for 9 days (S8NS9), non-shading for 8 days+shading for 9 days (NS8S9), and shading for 17 days (S17). Integrated solar radiation in NS was highest, and in S8NS9>NS8S9>S17 order. Integrated temperature did not shown difference among the treatments for growing periods. Growth and chlorophyll content of baby leaf vegetables were lowest in shading treatment for 8days. The highest growth characteristics (plant height, leaf length, leaf width, stem diameters, and fresh weight) and chlorophyll content of baby leaf vegetables was shown in NS. Increment rate of plant height and fresh weight of baby leaf vegetables were high in NS or S8NS9 as affected by light control for 8 days in early growth periods. These results suggest that we can utilize in low layer for early period and in high layer for harvest periods for baby vegetable production in rice seedling factory.

      • Association of Extracellular Cleavage of E-Cadherin Mediated by MMP-7 with HGF-Induced in vitro Invasion in Human Stomach Cancer Cells

        Lee, K.H.,Choi, E.Y.,Hyun, M.S.,Jang, B.I.,Kim, T.N.,Kim, S.W.,Song, S.K.,Kim, J.H.,Kim, J.-R. S. Karger AG 2007 European surgical research Vol.39 No.4

        <P><I>Background:</I> Proteolytic shedding of the ectodomain of a variety of transmembrane proteins, including cell-to-cell adhesion molecules, has been observed in solid cancers. We have investigated whether extracellular cleavage of E-cadherin mediated by matrix metalloproteinase-7 (MMP-7) is involved in hepatocyte growth factor (HGF) induced in vitro invasion in stomach cancer cells. <I>Methods:</I> The effects of HGF on the expression of E-cadherin/β-catenin and MMP-7 at both the protein and mRNA levels were assessed in stomach cancer cells, NUGC-3 and MKN-28, and in cells in which the expression of MMP-7 was downregulated by transfection with a MMP-7 short hairpin RNA plasmid. <I>Results:</I> Treatment with HGF increased the extracellular cleavage of E-cadherin and the release of MMP-7 and reduced the level of E-cadherin in a dose- and time-dependent manner. HGF treatment repressed the phosphorylation of β-catenin in a Triton-soluble fraction, but enhanced this phosphorylation in a Triton-insoluble fraction. The association of E-cadherin with β-catenin was decreased by HGF treatment in the Triton-soluble fraction. In addition, treatment of MMP-7 short hairpin RNA transfected NUGC-3 cells with HGF resulted in no extracellular cleavage of E-cadherin and also decreased the in vitro cell invasion. <I>Conclusions:</I> These results suggest that incubation with HGF mediated the release of MMP-7, resulting in extracellular cleavage of E-cadherin from stomach cancer cells. This might be a key mechanism in HGF-induced in vitro invasion and metastasis.</P><P>Copyright © 2007 S. Karger AG, Basel</P>

      • Characterization and expression analysis of a goose-type lysozyme from the rock bream Oplegnathus fasciatus, and antimicrobial activity of its recombinant protein

        Whang, I.,Lee, Y.,Lee, S.,Oh, M.J.,Jung, S.J.,Choi, C.Y.,Lee, W.S.,Kim, H.S.,Kim, S.J.,Lee, J. Academic Press 2011 Fish & shellfish immunology Vol.30 No.2

        Lysozyme (muramidase) represents an important defense molecule of the fish innate immune system. Known for its bactericidal properties, lysozyme catalyzes the hydrolysis of β-(1,4)-glycosidic bonds between the N-acetyl glucosamine and N-acetyl muramic acid in the peptidoglycan layer of bacterial cell walls. In this study, the complete coding sequence of a g-type lysozyme (RBgLyz) was identified in the Oplegnathus fasciatus rock bream fish genome by means of multi-tissue normalized cDNA pyrosequencing using Roche 454 GS-FLX(TM) technology. RBgLyz is composed of 669 bp, with a 567 bp open reading frame that encodes 188 amino acids. Protein motif searches indicated that RBgLyz contains the soluble lytic transglycosylase domain involved in maintaining cell wall integrity. Furthermore, RBgLyz shares significant identity (81.4%) with Chinese perch Siniperca chuatsi. Quantitative real-time RT-PCR analysis results showed that RBgLyz transcripts are constitutively expressed in various tissues from healthy rock breams. In order to determine RBgLyz function in immunity, its expression was analyzed in head kidney following exposure to known immune stimulants or pathogens. RBgLyz transcripts were significantly up-regulated in response to challenge with lipopolysaccharide (LPS) and Edwardsiella tarda, as compared to non-injected control fish. Polyinosinic:polycytidylic acid (poly I:C) dsRNA stimulated a moderate expression of RBgLyz, as did Streptococcus iniae but to a lesser extent. There were no specific time-dependent effects on RBgLyz mRNA expression observed in response to rock bream iridovirus (RBIV) infection. Taken together, the gene expression results indicated that g-type lysozyme plays a role in the innate immune response to LPS, poly I:C, E. tarda and S. iniae in rock bream. Thus, we generated recombinant RBgLyz in an Escherichia coli expression system and characterized its antimicrobial activity. Our results indicated that recombinant RBgLyz had lytic activity against Gram-negative Vibrio salmonicida, Gram-positive Listeria monocytogenes, S. iniae and Micrococcus lysodeikticus. In addition, observations by scanning electron microscope (SEM) confirmed that the cell morphology of M. lysodeikticus was altered in the presence of recombinant RBgLyz.

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