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>

Trypsin inhibitor 결여 大豆品種의 탐색 및 그의 遺傳育種學的 硏究 : I. Trypsin inhibitors의 전기영동 감정방법에 의한 대두 품종별 비교 및 DEAE-cellulose에 의한 분리 I. Soybean trypsin inhibitors: electrophoretic differences among varieties and their fractionation on DEAE-cellulose

金秀一,李錫河,李弘石,文亢植,羅志英 서울大學校 農科大學 1985 서울대농학연구지 Vol.10 No.1

대두의 단백추출액을 polyacrylamide gel 전기영동에 의하여 분류하고 trypsin inhibitor (T.I) band를 동정하였다. T.I. band는 전기영동한 gel을 trypsin으로 가수분해하거나 추출액에 trypsin을 처리한 후 전기영동하거나 발색기질을 이용하여 gel을 착색시키거나 또는 gel slice의 T.I.activity를 측정하는 등 네 가지 방법을 사용하여 검정하였다. 이중 추출액을 trypsin으로 처리한 후 전기영동하는 방법과 gel slice의 T.I.activity를 측정하는 방법이 가장 적합하였으며 두 방법의 결과를 비교하여 T.I.band를 검정하는 것이 보다 확실하였다. Sephadex G-75 Chromatography 에서 물로 추출한 대두 단백질은 3 fraction으로 분리하였고 T.I.activity는 제 2 fraction 에만 나타났다. Kunitz 및 Bowman-Birk형 inhibitor는 DEAE-cellulose column chromatography로 분리하였다. Kunitz형은 5개의 fraction으로, Bowman-birk형은 4개의 fraction으로 분리되었다. 단백질 추출액과 DEAE-cellulose chormatography에서 분리된 Kuniz 및 Bowman-Birk T.I.의 polyacryamide gel 전기영동 pattern을 비교하여 본 결과, 확실하게 동정된 T.I.band는 band3과 band4로서 각각 Orf등이 발표한 Ti¹과 Ti2에 해당하였으며, 그 외에 band 6과 band 10이 T.I.로 추정되었고 band 1,2,5,7,8,9는 T.I.가 아닌 것으로 판명되었다. trypsin inhibitor 함유량은 총 trypsin units inhibited 값(T.U.I)으로 볼 때 42품종에서 25에서 76까지 품종 간에 차이가 현저하였으며 시비 및 파종기의 영향은 나타나지 않았다. Ti¹ inhibitor 를 보유하고 있는 것은 37품종이었고, Ti²를 보유하는 것은 7품종이었으며, Ti¹과 Ti²를 같이 가지고 있는 품종은 발견되지 않았다. 이러한 품종의 Ti¹,Ti² 보유 pattern은 재배조건에 의해 변화되지 않았다. 2조합의 pattern은 재배조건에 의해 변화되지 않았다. 2조합의 정역교배에서 얻은 F₁ 종자의 전기영동 pattern을 비교해 본 결과, Ti¹품종끼리의 교배종자에서는 정역교배에 상관없이 Ti¹ inhibitor 만 나타났고 Ti¹품종과 Ti²품종의 교배종자에서는 Ti²를 모본으로 한 종자에서는 Ti¹과 Ti² 두 inhibitor가 검출되었으나 여교배에서는 모본의 Ti¹ inhibitor만 검출되었다. 여교배에서 Ti¹만 나타난 것은 분석시료 종자가 적었고 교배의 여부를 확인할 수 없어 모본의 세포질적 영향에 의한 것인지 또는 자가수정에 의한 것인지 분명치 않았다. The protein extracts from soybean seeds were examined by polyacrylamide gel electrophoresis and the trypsin inhibitor (T.I) bands were detected. The water-extractable protein was fractionated into three fractions by Sephadex G-75 gel filtration. The T.I activity was found only in the second fraction. Kunitz and Bowman-Birk inhibitors were fractionated by DEAE-cellulose chromatography into seven and six fractions, respectively. In kunitz inhibitor, 5 fractions were found to have T.I activity and 4 fractions in Bowman-Birk inhibitor. From the and patterns of the protein extracts and those of DEAE-cellulose chromatographic fractions, it was found that band 3 and 4 were T.I. band, corresponding to Ti¹ and Ti² band, respectively. In addition, band 6 and 10 were presumed to be T.I. band. Of the 42 varieties sampled, 35 revealed only Ti¹ band and 7 only Ti² band. The T.I. band patterns were not changed by the culture condition. The T.I. content, when expressed as the number of trypsin units inhibited (T.U.I), showed remarkable differences from 25 to 76 between varieties. The seedtime and fertilization condition had no effect on the T.I. content. Judged from the results of F ₁seeds analysis, we assumed that Ti¹ and Ti²band were controlled by codominant allele at a single locus.

Cytogenetic assessment of <i>Lilium longiflorum × L. hansonii</i> revealed by genomic in situ hybridization (GISH)

Mazharul, I.M.,Reshma, Y.,Jung, J.M.,Mohammad, D.D.,Lim, K.B. International Society for Horticultural Science 2019 Acta Horticulturae Vol.1237 No.-

<P> Martagon (<I>Lilium hansonii;</I> MM) and <I>Longiflorum</I> (LL) are two major groups under the family <I>Liliaceae</I>, used for modern breeding to introduce new inter-genomic lily cultivars. Interspecific F<SUB>1</SUB> hybrids (LM) introduced through cut-style method between two diploid <I>Lilium longiflorum (2n=2x=24)</I> and <I>Lilium hansonii (2n=2x=24)</I> were evaluated cytogenetically by genomic in situ hybridization (GISH) technique. However, GISH analysis of F<SUB>1</SUB> interspecific (LM) hybrids showed equal chromosomal contribution from both female <I>Lilium longiflorum</I> (LL) and male <I>Lilium hansonii</I> (MM). Each of the parent contributed 12 chromosomes except three crosses i.e., two of <I>L. longiflorum</I> 'White Tower' × <I>L. hansonii;</I> (2x-1) and one of <I>L. longiflorum</I> 'Bright Tower' × <I>L. hansonii;</I> (2x-1). Among 11 inter-genomic crosses, 3 crosses failed (False hybrid) and 8 crosses (True hybrids) showed different ploidy level i.e., 2n=2x=24, 2n=2x-1=23 and 2n=2x-1=23 respectively. Recombinant chromosome usually not found in F<SUB>1</SUB> interspecific lily hybrids. Most often, genomic recombination occurred in the cross between two genetically different parents. Chromosome pairing and crossing over normally occurred during meiosis in backcross progenies. However, in this study, genome analysis (GISH) of F<SUB>1</SUB> hybrids (<I>L. longiflorum</I> 'White Tower' × <I>L. hansonii</I>) showed four recombinant sites including two M/L and two L/M recombinant chromosomes that denotes high genetic relationship between <I>L. longiflorum</I> and <I>L. hansonii.</I> </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>

방사성 동위원소옥소(131I)에 의한 갑상선질환의 임상적 연구

이정상,이문호,고창순,노흥규,구인서,서환조,이경자,이홍규 대한핵의학회 1970 핵의학 분자영상 Vol.4 No.2

서울대학교 의과대학 내과학교실 및 방사성 동위원소 진료실에서 1960년 5월부터 1969년 10월까지 진료한 2,658명의 각종 갑상선 질환 환자에 대하여 131I에 의한 각종 갑상선 기능 검사 및 기능 항진증 환자에 대한 131I의 치료 성적을 종합 검토하여 아래와 같은 결론을 얻었다. 1) 2,658명의 갑상선 질환 환자중 독성 미만성 선종이 929명(34.9%)으로 가장 많고 비중독성 미만성 선종 및 비중독성 결절성 선종이 각각 762명(28.7%), 699명(26.3%)이며 기능저하가 210명(7.9%), 독성 결절성 선종이 58명(2.2%)였다. 2) 갑상선 질환의 성별 발생 빈도는 남자 300명(11.4%), 여자 2,358명(88.6%)로서 그 비는 1:8였다. 3) 연령별 발생 빈도는 20∼49세에서 전체의 79.1%인 2,102명이며 기능 항진증의 경우는 79.0%에 달하였다. 4) 각종 갑상선 기능 검사중 131I 섭취율, 131I 혈청내 방사능 BMR치등에 대한 고찰을 하는 한편 기능항진 및 저하증때 나타내는 각종 자학증세를 관찰하였다. 5) 갑상선 기능 항진증 환자 867명에 대하여 131I 치료를 하고 그 중 579명에서 47.8%의 초회 치료율을 확인하였다. 6) 131I 투여 후의 합병증인 기능 저하증의 발생 빈도는 초회 투여에서 6.75%였다. 7) 갑상선의 $quot; A summary of the clinical data of the (131)^I-thyroid function tests and the therapeutic results of 1(31I)^ among the 2,658 patients of various thyroid diseases treated over the past 10 years from May 1960 to Oct. 1969 at the Radioisotope Clinci and Laboratory, SNUH were presented and discussed. 1) The patients examined consisted of 929 cases (34.9%) of diffuse toxic goiter, 762 cases (28.7%) of diffuse nontoxic goiter, 699 cases (26.3%) of nodular nontoxic goiter, 58 cases (2.2%) of nodular toxic goiter and 210 cases (7.9%) of hypothyroidism. 2) There were 300 (11.4%) male and 2358 (88.6%) female, showing a ratio of 1:8. 3) The majority of patients (79.1%) were in the 3rd-5th decade of their lives. 4) The normal ranges, diagnostic values of (131)^I uptake test, 48 hrs serum activity, BMR and main subjective symptoms of various thyroid diseases were discussed. 5) In the 579 patients among 867 cases with hyperthyroidism treated with (131)^I, 47.8% were confirmed to be cured completely after single therapeutic doses. 6) The complications of 131I therapy were discussed and myxedema had developed in 6.75% of our patients. 7) The results of (131)^I thyroid function tests were analysed among the 160 cases of thyroid diseases which were confirmed the diagnosis with histopathological measures.

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>

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.

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>

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>

Identification of thioredoxin domain-containing protein 17 from big-belly seahorse <i>Hippocampus abdominalis</i>: Molecular insights, immune responses, and functional characterization

Liyanage, D.S.,Omeka, W.K.M.,Yang, Hyerim,Godahewa, G.I.,Kwon, Hyukjae,Nam, Bo-Hye,Lee, Jehee Elsevier 2019 Fish & shellfish immunology Vol.86 No.-

<P><B>Abstract</B></P> <P>Thioredoxin domain-containing protein 17 (TXNDC17) is a small protein (∼14 kDa) involved in maintaining cellular redox homeostasis via a thiol-disulfide reductase activity. In this study, TXNDC17 was identified and characterized from <I>Hippocampus abdominalis</I>. The open reading frame (ORF) consisted of 369 bp and 123 amino acids. Similar to the other thioredoxins, TXNDC17 contained a conserved WCXXC functional motif. The highest spatial mRNA expressions of <I>HaTXNDC17</I> were observed in the muscle, brain, and intestine. Interestingly, the mRNA expression of <I>HaTXNDC17</I> in blood showed significant upregulation at 48 h against all the pathogen associated molecular patterns (PAMPs) and bacteria. Further, <I>HaTXNDC17</I> transcripts in the trunk kidney were significantly upregulated at 24–48 h by bacterial endotoxin lipopolysaccharides (LPS), viral mimic polyinosinic: polycytidylic acid (poly I:C), and gram-negative bacteria (<I>Edwardsiella tarda</I>). The DPPH assay showed that the radical scavenging activity varies in a concentration-dependent manner. The insulin reduction assay demonstrated a significant logarithmic relationship with the concentration of rHaTXNDC17. Moreover, FHM cells treated with recombinant HaTXNDC17 significantly enhanced cellular viability under oxidative stress. Together, these results show that HaTXNDC17 function is important for maintaining cellular redox homeostasis and that it is also involved in the immune mechanism in seahorses.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Big-belly seahorse thioredoxin domain containing protein 17 maintain redox homeostasis. </LI> <LI> TXNDC17 is ubiquitously found in cytosol and extracellular space. </LI> <LI> Thiol active CXXC conserved motif consists in HaTXNDC17. </LI> <LI> Spatial and temporal mRNA expression was evaluated. </LI> <LI> Radical scavenging ability, antioxidant activity and cellular viability were measured. </LI> </UL> </P>