Molecular cloning and expression of cDNAs for two distinct granulocyte colony stimulating factor genes from black rockfish Sebastes schlegelii

Nam, B.H.,An, G.H.,Baeck, G.W.,Kim, M.C.,Kim, J.W.,Park, H.J.,Lee, D.C.,Park, C.I. Academic Press 2009 FISH AND SHELLFISH IMMUNOLOGY Vol.27 No.2

Granulocyte-colony stimulating factor (G-CSF) is a cytokine that stimulates the proliferation and differentiation of hematopoietic progenitor cells committed to the neutrophil/granulocyte lineage. Here, we report the two distinct granulocyte colony stimulating factor homologues from black rockfish Sebastes schlegelii. The G-CSF homologue cDNAs were isolated from the black rockfish LPS or Con A/PMA stimulated leukocyte cDNA libraries. The cDNA for the Black rockfish G-CSF-1 homologue predicts a peptide of 202 amino acids that is the closest to the Bastard halibut (Paralichthys olivaceus) G-CSF, whereas the cDNA of the Black rockfish G-CSF-2 homologue predicts a peptide of 212 amino acids that is the closest to the Fugu (Takifugu rubripes). In a healthy fish, the mRNAs of both G-CSF homologues were predominantly expressed in leukocytes, spleen, and gill. Expression of the black rockfish G-CSF-1 homologue was induced in peripheral blood leukocytes (PBLs) after stimulation with LPS, Con A/PMA, or Poly I:C, and G-CSF-2 homologue was strongly induced in PBLs after stimulation with Con A/PMA for 24 h only.

Purification and characterization of an antimicrobial histone H1-like protein and its gene from the testes of olive flounder, Paralichthys olivaceus

Nam, B.H.,Seo, J.K.,Go, H.J.,Lee, M.J.,Kim, Y.O.,Kim, D.G.,Lee, S.J.,Park, N.G. Academic Press 2012 Fish & Shellfish Immunology Vol.33 No.1

An approximately 21 kDa antimicrobial protein was purified from an acidified testis extract of olive flounder, Paralichthys olivaceus, by ion-exchange and C<SUB>18</SUB> reversed-phase HPLC. A comparison of the N-terminal amino acid sequence with those of other known antimicrobial polypeptides revealed high homology between this antimicrobial protein and other histone H1 molecules; thus, it was designated flounder histone H1-like protein (fH1LP). fH1LP showed potent antimicrobial activity against Gram-positive bacteria, including Bacillus subtilis, Staphylococcus aureus, and Streptococcus iniae (minimal effective concentrations [MECs], 2.8-30.0 μg/ml), Gram-negative bacteria, including Aeromonas hydrophila, Escherichia coli D31, Vibrio parahaemolyticus (MECs, 1.4-12.0 μg/ml), and Candida albicans (MEC, 2.0 μg/ml). cDNA cloning and tissue distribution studies of fH1LP indicated that it is constitutively expressed in testis and ovary. The fH1LP expression level was significantly dependent on developmental stage, and decreased dramatically after hatching. However, lipopolysaccharide stimulation did not induce fH1LP mRNA in other immune organs, including the kidney and spleen. These results suggest that fH1LP plays an important role in innate immunity in fish during reproduction, including mating, fertilization, and hatching.

밀링 가공 시 절삭력 신호를 이용한 공구 상태 모니터링에 관한 연구

양기동(G. D. Yang),남수현(S. H. Nam),박경희(K. H. Park) Korean Society for Precision Engineering 2021 한국정밀공학회 학술발표대회 논문집 Vol.2021 No.11월

밀링 가공시 공구의 마모 및 파손과 같은 이상상태는 가공 오차 및 불량품 발생 등으로 인한 생산성 저하의 원인이 된다. 이상상태 감지를 위해 공구동력계(Dynamometer), 음향 방출 센서(Acoustic Emission Sensor), 가속도 센서(Acceleration Sensor), 전류/전압 센서 등이 주로 사용되고 있다. 본 논문에서는 공구동력계 신호를 이용하여 공구 상태에 따른 절삭력 변화에 대해 분석하였다. 피삭재는 SM45C 이며, 공구는 엔드밀(D16 mm, Indexable Endmill, 2-flute)을 이용하였다. 가공 조건은 가공 속도 3,000-5,000 rpm, 가공 깊이 1-6 mm, 가공 너비 1-13 mm, 날당 이송 0.05-0.15 mm/tooth 등 다양한 조합으로 설정하였다. 각각의 가공 조건하에서 공구 수명(마모 기준: Fleank Wear 300 μm)에 도달할 때까지 실험을 진행하였다. 마모의 형태로는 여유면 마모, 노치 마모, 치핑/파손이 나타났다. 절삭력은 X/Y 축의 Peak, RMS (Root Mean Square), RMS 의 Moving Average 를 계산하여 가공 Pass 에 대한 대표값을 추출하였다. 점진적 공구 마모 발생시 가공 방향 절삭력보다 반경 방향 절삭력의 증가가 크게 나타났다. 공구 마모 증가에 따른 절삭력 변화(초기 절삭력 대비 증가율, 직전 패스에서의 절삭력 대비 증가율)를 분석하여 상관관계를 조사하였다. 두날 공구 기준 상태 모니터링을 위해 공구의 1 날이 파손된 경우 파손된 날의 절삭력이 급격히 감소하므로 두 날의 절삭력 Peak 값에 대한 차이로 검출 가능하다. 반면, 공구의 2 날이 파손된 경우 가공 방향의 절삭력이 크게 증가하는 것이 확인되었다. 향후, 실제 가공현장에서의 도입을 목표로 스핀들 진동/전류 신호를 통한 공구마모 검출에 대한 연구를 진행하고자 한다.

14.6 A GeV ^28Si 중이온이 원자핵건판내에서 발생시킨 핵반응에서 생성된 2차입자의 발생각 분포

김종오,김태연,남신우,신택수,우종관,이세병,임계엽,장세덕,조재희,천병구,임인택,김기영 慶尙大學校 기초과학연구소 1990 基礎科學硏究所報 Vol.6 No.-

14.6A GeV^28Si 중이온이 원자핵 건판내에서 발생시킨 N_h=1인 핵반응에서 생성된 47개의 파쇄 α 입자와 537개의 단일하전 2차입자의 발생각들을 측정하여 변수 exp(γ-η_b)의 포괄적 분포를 회귀함수 dN=exp[a+χ{exp(γ-η_b)d{exp(γ-η_b)}로 적합시켰다. 여기서 의사신속도 γ=arctanh(cosθ)=-ln tan(θ/2)이고, 입사 중이온의 신속도 η_b=3.445이다. 그 적합결과 파쇄 α입자의 경우 χ=-0.052±0.011이고, 파쇄 p입자의 경우 χ=-0.141±0.015이었다. For LS emission angles of 47 α fragments and 537 single-charged shower particles, produced by the N_h (the number of heavyprongs)=1 interactions of 14.6 A GeV^28Si nuclei in the nuclear emulsion, the distribution of the parameter exp(γ-η_b) is well expressed by dN=exp[a+χ{exp(γ-η_b)d{exp(γ-η_b)}with χ=-0.052±0.011 for αfragments and χ=-0.141±0.015 for p 'fragments', where the pseudorapidity of secondaries γ=arctanh(cosθ)=-ln tan(θ/2) and the rapidity of incident heavy ions, η_b=3.445.

Chronic inflammatory demyelinating polyradiculoneuropathy in children: characterized by subacute, predominantly motor dominant polyeuropathy with a favorable response to the treatment

Jo, H. Y.,Park, M.-G.,Kim, D.-S.,Nam, S.-O.,Park, K.-H. Blackwell Publishing Ltd 2010 Acta neurologica Scandinavica Vol.121 No.5

<P>Jo HY, Park M-G, Kim D-S, Nam S-O, Park K-H. Chronic inflammatory demyelinating polyradiculoneuropathy in children: characterized by subacute, predominantly motor dominant polyeuropathy with a favorable response to the treatment.Acta Neurol Scand: 2010: 121: 342–347.© 2009 The Authors Journal compilation © 2009 Blackwell Munksgaard.</P><P>Objectives – </P><P>Chronic inflammatory demyelinating polyradiculopathy (CIDP) is less well-studied in children than in adults, probably due to its relative rarity. This study was performed in order to characterize the clinical features of CIDP in children.</P><P>Materials and methods – </P><P>Twenty-eight patients with CIDP who were followed up for more than 1 year were included, and were divided into a child (<I>n</I> = 7, age <16) and an adult group (<I>n</I> = 21, age ≥16). Then, we have assessed the initial progression pattern, clinical course, and serial nerve conduction findings in each patient. Finally, differential features in child and adult group were analyzed.</P><P>Results – </P><P>Distinguishing features in the child group include subacute progression within less than 2 months, predominant motor system involvement in lower extremities, and marked improvement in response to immune modulating therapy. Our study also suggested that serial nerve conduction study may be useful in assessing the effectiveness of the treatment in children. </P><P>Conclusions – </P><P>Our study showed that children with CIDP have some distinguishing features from adults in terms of clinical course and response to treatment.</P>

In vitro synergistic anticancer activity of the combination of T-type calcium channel blocker and chemotherapeutic agent in A549 cells

Byun, J.S.,Sohn, J.M.,Leem, D.G.,Park, B.,Nam, J.H.,Shin, D.H.,Shin, J.S.,Kim, H.J.,Lee, K.T.,Lee, J.Y. Pergamon Press 2016 Bioorganic & medicinal chemistry letters Vol.26 No.3

As a result of our continuous research, new 3,4-dihydroquinazoline derivative containing ureido group, KCP10043F was synthesized and evaluated for T-type Ca<SUP>2+</SUP> channel (Ca<SUB>v</SUB>3.1) blockade, cytotoxicity, and cell cycle arrest against human non-small cell lung (A549) cells. KCP10043F showed both weaker T-type Ca<SUP>2+</SUP> channel blocking activity and less cytotoxicity against A549 cells than parent compound KYS05090S [4-(benzylcarbamoylmethyl)-3-(4-biphenylyl)-2-(N,N',N'-trimethyl-1,5-pentanediamino)-3,4-dihydroquinazoline 2 hydrochloride], but it exhibited more potent G<SUB>1</SUB>-phase arrest than KYS05090S in A549 cells. This was found to be accompanied by the downregulations of cyclin-dependent kinase (CDK) 2, CDK4, CDK6, cyclin D2, cyclin D3, and cyclin E at the protein levels. However, p27<SUP>KIP1</SUP> as a CDK inhibitor was gradually upregulated at the protein levels and increased recruitment to CDK2, CDK4 and CDK6 after KCP10043F treatment. Based on the strong G<SUB>1</SUB>-phase cell cycle arrest of KCP10043F in A549 cells, the combination of KCP10043F with etoposide (or cisplatin) resulted in a synergistic cell death (combination index=0.2-0.8) via the induction of apoptosis compared with either agent alone. Taken together with these overall results and the favorable in vitro ADME (absorption, distribution, metabolism, and excretion) profiles of KCP10043F, therefore, it could be used as a potential agent for the combination therapy on human lung cancer.

Purification and antimicrobial function of ubiquitin isolated from the gill of Pacific oyster, Crassostrea gigas

Seo, J.K.,Lee, M.J.,Go, H.J.,Kim, G.D.,Jeong, H.D.,Nam, B.H.,Park, N.G. Pergamon Press 2013 Molecular immunology Vol.53 No.1

An antimicrobial polypeptide was purified from an acidified gill extract of Pacific oyster (Crassostrea gigas) by C<SUB>18</SUB> reversed-phase HPLC. The purified polypeptide had a molecular weight of 8471Da containing 74 amino acid residues. Comparison of the obtained N-terminal sequences with those of others revealed that it was identical to ubiquitin reported from other species and named cgUbiquitin. cgUbiquitin showed broad potent antimicrobial activity against Gram-positive and -negative bacteria including Streptococcus iniae and Vibrio parahemolyticus (minimal effective concentrations, 7.8 and 9.8μg/mL), respectively, without hemolytic activity. The cgUbiquitin cDNA was identified from an expressed sequence tag (EST) library of oyster gill as a precursor form, encoding ubiquitin consisting of 76 amino acids fused to ribosomal protein of S27. Although the cgUbiquitin precursor mRNA was expressed at the intermediate level in the gill, the mRNA was significantly up-regulated at 48h post injection with Vibrio sp. Analysis of the cgUbiquitin C-terminus by carboxypeptidase B treatment and comparison of the retention times revealed that cgUbiquitin lacks the terminal Gly-Gly doublet and ends in an C-terminal Arg residue which might be related to antimicrobial activity. Study of the kinetics of killing and membrane permeabilization showed that this peptide was not membrane permeable and acted through a bacteriostatic process. According to the homology modeling, this peptide is composed of three secondary structural motifs including three α-helices and four β-strands separated by 7 loops regions. Our results indicate that cgUbiquitin might be related to the innate immune defenses in the Pacific oyster and this is the first report for antimicrobial function of ubiquitin isolated from any oyster species.

추간판내 유전자 치료 : 퇴행성 질환에 있어 치료의 의의 Therapeutic Implication in Degenerative Disc Disease

문성환,박문수,강용호,김형중,이환모,김남현,Gillbertson, Lars G.,Kang, James D. 대한척추외과학회 2000 대한척추외과학회지 Vol.7 No.4

연구계획 : 인간의 추간판 세포와 가토의 추간판 조직을 이용하여 시험관내 및 생체내 실험적 연구를 시행하였다. 목 적 : 3차원적으로 배양된 인간의 추간판 세포와 생체내의 가토의 추간판에 치료적 유전자를 전달하고 기질 생성 효과가 있는지를 알아보는 것이다. 대상 및 방법 : 12례의 추간판 술실에서 채취된 추간판 조직에서 세포를 유리하고 이를 3차원적으로 배양한 후 Ad/TGF-β1으로 감염시켰다. 15마리의 가토를 사용하여 전방 도달법으로 추간판을 노출시키고 Ad/TGF-β을 주입한 다음 6주 후 희생시켰다. 배양약의 1Ad/TGF-β1농도는 ELISA법으로 측정하였고 새로이 생성된 당단백은 35S-sulfate을 이용한 Sephadex G-25PD-10 column chromatography 법으로 측정하였다. 결과 : Ad/TGF-β1로 감염된 인간의 추간판 세포에서 TGF-β1생성은 증가하였고 Ad/TGF-β1로 감염된 가토의 추간판 조직은 대조군에 비해 200%의 TGF-β1생성 증가가 있었다. Ad/TGF-β1로 감염된 인간의 추간판 세포에서 290%의 당단백 생성 증가가 있었고(p<0.05) Ad/TGF-β1로 감염된 가토의 추간판 조직은 대조군에 비해 85%의 당단백 생성 증가가 있었다(p<0.05). 결론 : 시험관 및 생체내 실험에서 유전자 전달군은 대조군에 비해 의의 있는 당단백 생성 증가가 있었는데 이는 아데노바이러스를 이용한 추간판내 유저자 전달법이 추간판 퇴행의 치료와 예방에 적용 가능한 접근법임을 시사한다. Study Design : In vitro and in vivo studies to determine the anabolic effects of entervertebral desc (IVD) to adenovirus-mediated therapeutic gene transfer. Objectives : To quantify the anabolic effect of human IVD cells in vitro and rabbit IVD in vivo to therapeutic gene transfer. Sumary of Literature Review : An alternative possibility to delivery of growth factors, in continuous manner, is the genetic modification of disc cells through gene transfer. Contemplating to extend this approach to treatment of disc degeneration, it is neccessary to demonstrate anabolic effect of human IVD cells and rabbit disc to therapeutic gene transfer. Materials and Methods : In vitro:IVD tissue was obtained from twelve patients. IVD cells were then isolated, cultured, and transduced with Ad/TGF-β1. Genetically modified disc cells were incorporated into alginate beads and cultured. In vivo:Fifteen skeletally mature New Zealand white rabbit were used. 15ul of saline containing Ad/TGF-β1 were injected into the nucleus pulposus of the disc in six rabbits. All rabbits were sacrificed 6 weeks after surgery. Nucleus pulposus tissues were harvested, weighted, and cultured. Conditioned medium of alginate bead and rabbit disc tissue cultures were subjected to ELISA to detect TGF-β1 production. Newly synthesized proteoglycan were analyzed using chromatography on Sephadex G-25 in PD-10 columns after S35-sulfatged incorporation. Results : Concentration of TGF-β1 increased over time in alginate beads cultures transduced with Ad/TGF-β1. At 6 weeks nucleus pulposus tissue from the disc injected with Ad/TGF-β1 exhibited 200%(p<0.05) increase in TGF-β1 production. There was statistically significant 290% increase in newly synthesizd\ed proteoglycan in alginate cultures transduced with Ad/TGF-β1 (p<0.05) compared to control. At 6 weeks nucleus pulposus tissue from the disc injected with Ad/TGF-β1 exhibited 85% increase in proteoglyan synthesis (p<0.05) over that of intact control. Conclusion : In this study, we observed the robust upregulation of proteoglycan synthesis in gene transferred disc cells in vitro and in vivo-indicating good prospects for biologic effects of therapeutic gene therapy in the disc using adenovirus-mediated approach.

HIV env 유전자의 V1-V5 부위 분석에 의한 HIV 분리주의 subtype 및 변이 분석

김성순,강춘,박근용,남정구,최병선,기미경,서순덕,이성래,이주실 국립보건연구원 1999 국립보건원보 Vol.36 No.-

LiF:Mg,Cu,Na,Si의 열발광 피크 분석

남영미,정운혁,이대원,도시홍,이종환,김기동 동의대학교 기초과학연구소 1998 基礎科學硏究論文集 Vol.8 No.1

Five different methods were used to determine the trap depth E, the frequency factor s, and the kinetic order associated with the glow peaks in a LiF:Mg, Cu, Na, Si phosphor, which had been newly developed indigenously. The methods used were the initial-rise method, the total glow-peak method, the peak-shaped method, the variable heating-rate method, and the isothermal decay method. The peaks obeyed simple first-order kinetics following the model of Randal and Wilkins. The values of the trap depth E determined for peaks 1 to 4 were 0.69∼0.95, 0.71∼1.17, 1.77∼1.95 and 0.66∼1.22 eV with frequency-factor, s, values of 10^8∼10^11, 10^9∼10^11, 10^19∼10^20 and 10^8∼10^10 s^-1, respectively.