Cloning and functional characterization of the p65 subunit of NF-κB from olive flounder (Paralichthys olivaceus)

Kong, H.J.,Moon, J.H.,Moon, J.Y.,Kim, J.M.,Nam, B.H.,Kim, Y.O.,Kim, W.J.,Lee, S.J. Academic Press 2011 Fish & shellfish immunology Vol.30 No.1

NF-κB is a master transcription factor found in almost all cell types that responds to diverse cellular stimuli by activating the expression of stress response genes, including immune-related genes. cDNA encoding the p65 subunit of olive flounder (Paralichthys olivaceus) NF-κB (Po-p65) was isolated through an EST analysis of an olive flounder cDNA library, a screen of BAC library, and rapid amplification of cDNA ends (RACE). The cDNA for Po-p65 encodes a polypeptide 626 amino acids in length containing a well-conserved Rel-homology domain (RHD). The primary sequence of Po-p65 showed strong homology with p65 from perch and zebrafish (82.7 and 64.4%, respectively), and shared 43.4-42.1% homology with p65 from other species, including mammals, while the N-terminal RHD of Po-p65 showed strong identity (95.6-67.8%) with that of other species. Po-p65 mRNA expression was detected in all flounder tissues examined. The over-expression of full-length Po-p65 (Po-p65f), but not of a Po-p65 C-terminus deletion mutant (Po-p65ΔC), stimulated κB element-driven reporter (κB-luc) activity in a dose-dependent manner and regulated the expression of p65 target genes, including TNF-α and IκB-α, in HINAE olive flounder cells. Po-p65f translocated to the nucleus following stimulation with poly I:C in HINAE cells. Together, these results suggest that Po-p65 is evolutionarily and functionally conserved in flounder and mammals and may provide clues to the detailed molecular mechanism(s) underlying immune response regulation in flounder.

<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>

Early Regulation of Viral Infection Reduces Inflammation and Rescues Mx-positive Mice from Lethal avian Influenza Infection

Song, M.S.,Cho, Y.H.,Park, S.J.,Pascua, P.N.Q.,Baek, Y.H.,Kwon, H.I.,Lee, O.J.,Kong, B.W.,Kim, H.,Shin, E.C.,Kim, C.J.,Choi, Y.K. American Association of Pathologists and Bacteriol 2013 The American journal of pathology Vol.182 No.4

Differing sensitivity of influenza A viruses to antiviral effects of the Myxovirus resistance (Mx) protein implies varying global gene expression profiles in the host. The role of Mx protein during lethal avian influenza (AI) virus infection was examined using Mx1-deficient C57BL/6 (B6-Mx1<SUP>-/-</SUP>) and congenic Mx1-expressing (B6-Mx1<SUP>+/+</SUP>) mice infected with a virulent, mouse-adapted avian H5N2 Ab/Korea/ma81/07 (Av/ma81) virus. After infection, B6-Mx1<SUP>+/+</SUP> mice were completely protected from lethal AI-induced mortality, and exhibited attenuated clinical disease and reduced viral titers and pathology in the lungs, compared with B6-Mx1<SUP>-/-</SUP> mice. Transcriptional profiling of lung tissues revealed that most of the genes up-regulated after infection are involved in activation of the immune response and host defense. Notably, more abundant and sustained expression of cytokine/chemokine genes was observed up to 3 dpi in B6-Mx1<SUP>-/-</SUP> mice, and this was associated with excessive induction of cytokines and chemokines. Consequently, massive infiltration of macrophages/monocytes and granulocytes into lung resulted in severe viral pneumonia and potentially contributed to decreased survival of B6-Mx1<SUP>-/-</SUP> mice. Taken together, our data show that dysregulated gene transcriptional activity corresponded to persistent induction of cytokine/chemokines and recruitment of cytokine-producing cells that promote inflammation in B6-Mx1<SUP>-/-</SUP> mouse lungs. Thus, we provide additional evidence of the interplay of genetic, molecular, and cellular correlates governed by the Mx1 protein that critically determine disease outcome during lethal AI virus infection.

한국 재래 닭 품종 특성 및 초기성장 개량을 위한 분자표지 개발

오재돈,박미현,공홍식,이학교,전광주,연성흠,상병돈,최철환,조병욱,Oh J. D.,Park M. H.,Kong H. S.,Lee H. K.,Jeon G. J.,Yeon S. H.,Sang B. D.,Choi C. H.,Cho B. W. 한국가금학회 2005 韓國家禽學會誌 Vol.32 No.1

This study was conducted to estimate the effects of genotype for chicken major histocompatibility complex (MHC) B-LB genes on economic traits. To detect polymorphism, 400 bp fragments of MHC B-LB genes were obtained and sequenced. After digestions using restriction enzyme Hea III, two restriction enzyme sites were observed. There were two mutations at position 427 and 651 those were decided as Type I and Type II, respectively. Using RFLP analyses, type I were genotyped to TT, TC and CC, and type II to MM, Mm and mm. The relatively higher TC genotype frequencies (0.8) of Type I and Mm genotype frequencies (0.88) of Type II were observed in Korean native chickens. The effects of the genotype on 150 days body weight trait were investigated by the associations of CC and Mm genotypes (P<0.05) in Korean native chickens. This result suggests that a significant association exists between the SNP and 150 days body weight. 본 연구는 한국 재래 닭의 유전적 특성을 분자표지를 이용하여 그 차이를 규명하고 초기성장에 미치는 영향을 분석하여 이를 이용한 재래닭의 개량을 목적으로 실시하였다. MHC class II B-LB 유전자 내의 염기변이체가 경제형질에 미치는 영향에 대하여 연구하였다. MHC class II B-LB유전자 내 400 bp 크기의 유전자를 증폭하여 염기서열 분석과 제한효소 처리를 이용한 다형성 분석을 실시하였다. 연구결과 두 개의 제한효소 절단지역이 발견되었으며 427 지역을 Type I 으로 651 지역은 Type II로 정하여 RFLP 분석을 실시하였다. Type I지역의 유전자형은 TT, TC, CC로 나타났으며, TypeII 지역의 유전자형은 MM, Mm, mm으로 나타났다. TC와 Mm 유전자형이 다른 유전자형과 비교하였을 때 한국재래 닭에서 높은 출현빈도를 보였다(0.8, 0.88). 유전자형이 한국 재래 닭의 150일령 체중에 미치는 영향을 분석한 결과 CC와 Mm 유전자형에서 통계적 유의성이 도출되었다 (P<0.05). 따라서 본 연구의 결과를 이용하여 한국 재래 닭의 유전적 특성을 규명할 수 있으며 초기 성장이 높은 성적을 나타내는 CC, Mm 유전자형을 개량에 이용하게 된다면 큰 효과를 얻을 수 있을 것으로 사료되어진다. 본 연구의 결과는 차후 한국 재래 닭의 과학적이고 지속적인 유전자원의 보존과 육종 전략에 있어 매우 유용하게 활용될 것으로 기대된다.

Two novel phospholipid hydroperoxide glutathione peroxidase genes of <i>Paragonimus westermani</i> induced by oxidative stress

KIM, S.-H.,CAI, G.-B.,BAE, Y.-A.,LEE, E.-G.,LEE, Y.-S.,KONG, Y. Cambridge University Press 2009 Parasitology Vol.136 No.5

<B>SUMMARY</B><P>Phospholipid hydroperoxide glutathione peroxidase (PHGPx; GPx4) plays unique roles in the protection of cells against oxidative stress by catalysing reduction of lipid hydroperoxides. We characterized 2 novel <I>GPx</I> genes from a lung fluke, <I>Paragonimus westermani</I> (designated <I>PwGPx1</I> and <I>PwGPx2</I>). These single copy genes spanned 6559 and 12 371 bp, respectively, and contained each of 5 intervening introns. The <I>PwGPx2</I> harboured a codon for Sec and a Sec insertion sequence motif. Proteins encoded by the <I>Paragonimus</I> genes demonstrated a primary structure characteristic to the PHGPx family, including preservation of catalytic and glutathione-binding domains and absence of the subunit interaction domain. Expression of <I>PwGPx1</I> increased gradually as the parasite matured, whereas that of <I>PwGPx2</I> was temporally regulated. <I>PwGPx2</I> was expressed at the basal level from the metacercariae to the 3-week-old juveniles; however, the expression was significantly induced in the 7-week-old immature worms and reached a plateau in the 12-week-old adults and eggs. PwGPx1 and PwGPx2 were largely localized in vitellocytes within vitelline glands and eggs. Oxidative stress-inducible paraquat, juglone and H2O2 substantially augmented the <I>PwGPx1</I> and <I>PwGPx2</I> expressions in viable worms by 1·5- to 11-fold. Our results strongly suggested that PwGPxs may actively participate in detoxification of oxidative hazards in <I>P. westermani</I>.</P>

Selective novel inverse agonists for human GPR43 augment GLP-1 secretion

Park, B.O.,Kim, S.H.,Kong, G.Y.,Kim, D.H.,Kwon, M.S.,Lee, S.U.,Kim, M.O.,Cho, S.,Lee, S.,Lee, H.J.,Han, S.B.,Kwak, Y.S.,Lee, S.B.,Kim, S. North-Holland ; Elsevier Science Ltd 2016 european journal of pharmacology Vol.771 No.-

<P>GPR43/Free Fatty Acid Receptor 2 (FFAR2) is known to be activated by short-chain fatty acids and be coupled to G(i), and G(q), family of heterotrimeric G proteins. GPR43 is mainly expressed in neutrophils, adipocytes and enteroendocrine cells, implicated to be involved in inflammation, obesity and type 2 diabetes. However, several groups have reported the contradictory data about the physiological functions of GPR43, so that its roles in vivo remain unclear. Here, we demonstrate that a novel compound of pyrimidinecarboxamide class named as BTI-A-404 is a selective and potent competitive inverse agonist of human GPR43, but not the murine ortholog. Through structure-activity relationship (SAR), we also found active compound named as BTI-A-292. These regulators increased the cyclic AMP level and reduced acetate-induced cytoplasmic Ca2+ level. Furthermore, we show that they modulated the downstream signaling pathways of GPR43, such as ERK, p38 MAPK, and NF-kappa B. It was surprising that two compounds augmented the secretion of glucagon-like peptide 1 (GLP-1) in NCI-H716 cell line. Collectively, these novel and specific competitive inhibitors regulate all aspects of GPR43 signaling and the results underscore the therapeutic potential of them. (C) 2015 Elsevier B.V. All rights reserved.</P>

NFATc4 and ATF3 Negatively Regulate Adiponectin Gene Expression in 3T3-L1 Adipocytes

Kim, H. B.,Kong, M.,Kim, T. M.,Suh, Y. H.,Kim, W.-H.,Lim, J. H.,Song, J. H.,Jung, M. H. American Diabetes Association 2006 Diabetes Vol.55 No.5

<P>Expression of adiponectin decreases with obesity and insulin resistance. At present, the mechanisms responsible for negatively regulating adiponectin expression in adipocytes are poorly understood. In this investigation, we analyzed the effects of 5' serial deletion constructs on the murine adiponectin promoter. Here, we identified the repressor region located between -472 and -313 bp of the promoter. Removal of the putative nuclear factor of activated T-cells (NFATs) binding site increased the promoter activity, and overexpression of NFATc4 reduced the promoter activity. Treatment with the calcium ionophore A23187, an activator of NFAT, reduced mRNA as well as promoter activity. The binding of NFATc4 to the promoter was associated with increased recruitment of histone deacetylase 1 and reduced acetylation of histone H3 at the promoter site. In addition, binding of activating transcription factor 3 (ATF3) to the putative activator protein-1 site located adjacent to the NFAT binding site also repressed the promoter activity. Treatment with thapsigargin, an inducer of ATF3, reduced both mRNA and promoter activity. Importantly, the binding activities of NFATc4 and ATF3, increased significantly in white adipose tissues of ob/ob and db/db mice compared with controls. Taken together, this study demonstrates for the first time that NFATc4 and ATF3 function as negative regulators of adiponectin gene expression, which may play critical roles in downregulating adiponectin expression in obesity and type 2 diabetes.</P>

High-speed column-line driving with polarity switch-embedded output buffer amplifiers for TFT-LCD application

An, C.-H,Ko, J.-H,Kim, H.-R,Kong, B.-S IET 2015 Electronics letters Vol.51 No.1

<P>A novel high-speed column-line driving scheme having output buffer amplifiers embedded with polarity multiplexer switches is proposed for use in large-sized thin-film transistor liquid-crystal displays. The proposed driving scheme does not have explicit output-polarity switches, resulting in lower settling time. Experimental results in a 1.2 μm 13.5 V CMOS process indicated that using the proposed driving scheme the settling times to reach 99% of target voltages for the dot and column inversions were improved by up to 48.6%. This driving scheme can be applied to class AB- or class B-type amplifiers for liquid-crystal display column drivers and output buffers controlled by output switches.</P>

한우 경제형질에 미치는 Mitochondrial DNA D-loop 영역의 염기서열 변이효과

오재돈,윤두학,공홍식,임현진,이학교,조병욱,홍기창,전광주 한국동물자원과학회 2003 한국축산학회지 Vol.45 No.6

This study was performed to analyse the sequences of variations of mtDNA D-loop and their effects on carcass traits in Hnawoo(Korean cattle). The resulting sequences were compared with perviously published sequences for other cattle breeds(GenBank J01394). The PCR was used to amplify a total of 964 bp between nucleotide 15758 and 383 within D-loop region of mtDNA using specific primers. Twenty five polymorphic sites by nucleotide substitution were found in mtDNA of Hanwoo. The frequencies of positions at 169, 16042, 16093. 16119, 16255 and 16302 nt with high levels of sequence polymorphism were 0.891, 0.117, 0.109, 0.182, 0.197 and 0.117, respectively. The substitution effect at 169 and 16119 nt was found significant on marbling score. Also substitution effect at 169 and 16042 nt was highly significant(p<0.01) on backfat, thickness. Polymorphisrn of mtDNA sequence in D-loop region could be useful for the analysis of cytoplasmic genetic variation and associations with the other economically important traits and maternal lineage analysis in Hanwoo.

흑연료 원자흡수 분광법에 의한 혈중의 납, 카드뮴 정량을 위한 외부정도관리 시료제조 및 분석

이공주,임홍빈 이화여자대학교 생명과학연구소 1995 생명과학연구논문집 Vol.6 No.-

납과 카드뮴을 포함하는 여러 가지 농도의 동결건조된 혈액이 외부정도관리 시료로서 제조되었다. 이 시료들은 흑연료 원자흡수분광법(GFAAS)을 이용하여 성능이 파악되었다. 매트릭스 개선제로서 0.1% ammonium dihydrogen phosphate와 0.1% Triton X-100을 사용하여 섭씨 600내지 650도의 희화온도에서 혈액에 있는 납과 카드뮴의 정량 분석을 위한 GFAAS의 최적 분석조건이 얻어졌다. 제조된 혈액의 균질도와 안정도는 최적화된 분석조건에서 연구되었다. Lyophilized whole blood samples containing various concentrations of Pb and Cd have been prepared as external quality control materials. These materials have been characterized with graphite furnace atomic absorption spectrometry(GFAAS). The optimized obtained at the ashing temperature of 600~650℃ with 0.1% ammonium dihydrogen phosphate and 0.1% Triton X-100 as matrix modifier. Homogeniety and stability of the prepared whole blood have been studied at the optimized analytical condition.