소 모색관련 유전자 MC1R 의 RCR - RFLP Marker 를 이용한 한우육 판별

정의룡,김우태,김연수,한상기 한국동물자원과학회 2000 한국축산학회지 Vol.42 No.4

The melanocortin 1 receptor(MClR) plays a central role in regulation of eumelanin(black/brown) and phaeomelanin(red/yellow) pigment synthesis within the mammalian melanocytes. Mutations within the gene encoding MC1R have been shown to explain coat colour variations within several species including cattle. This study was performed to develop the identification technique of Hanwoo meat using MC1R gene associated with the coat colors of cattle. Alleles of the MC1R locus were detected by PCR-RFLP analysis and genotype frequency and DNA sequences of MC1R gene were compared among cattle breeds. Genomic DNA was extracted from meat or blood samples of five breeds including Hanwoo(n=200), Holstein(n=100), Angus(n=20), Hereford(n=20) and Charolais(n=20). The MC1R gene was used to amplify 739bp and 173bp of the bovine E-locus corresponding to positions 228-966 and 318-490, respectively, using the two specific primers. The amplified products were digested with Bse118 I or Msp I and Aci I enzymes, and DNA fragments were separated by gel electrophoresis for RFLP genotype analysis. Six genotypes, E^D/E^D E^D/E^+, E^D/e, E^+/E^+,E^+/e and e/e, controlled by three alleles E^D, E^+ and e were observed in MC1 locus. When the amplified DNA product(739bp) was digested with Bse118 I enzyme, Hanwoo meat showed a single band of 739bp, whereas two fragments of 531bp and 208bp were detected in Holstein meat and Angus breed, respectively. Also, in the RFLP patterns using Msp I enzyme, Hanwoo meat produced two fragments of 535bp and 174bp, while three fragments of 328bp, 207bp and 174bp were observed in Holstein meat and Angus breeds, respectively. Therefore, breed-specific RFLP markers showing distinct differences between these breeds were found by PCR-RFLP analysis. When the amplified DNA product(173bp) was digested with Aci I enzyme to classify subtype of E allele, the E^D allele produced three fragments of 97, 68 and 8bp, while the E^+ and d alleles produced two fragments of 173 and 8bp according to the Aci I recognition sequence. Among the six genotypes, two genotypes of E^+/e and e/e were observed in Hanwoo and their frequencies were 0.07 and 0.93, respectively. However, the E^D/ED and E^D/e genotypes were present in Holstein and E^D/E^D, E^D/E^+ and E^D/e genotypes in Angus breeds. Therefore, the E^+/e and e/e genotypes observed in Hanwoo and E^D/E^D, E^D/E^+ and E^D/e genotypes detected only in Holstein and Angus breeds may be useful as breed-specific DNA markers for distinguishing between Hanwoo meat and Holstein and Angus meats. When comparing MC1R sequences among Hanwoo, Holstein and Angus, a Gly → Val amino acid change due to a single base(G) deletion at colon 104 was found in Hanwoo. Consequently, breed specific RFLP genotypes of MC1R gene related to bovine coat colors could be used as DNA markers for identification of Hanwoo meat from Holstein and Angus meats.

The E3 ubiquitin ligase CHIP selectively regulates mutant epidermal growth factor receptor by ubiquitination and degradation

Chung, C.,Yoo, G.,Kim, T.,Lee, D.,Lee, C.S.,Cha, H.R.,Park, Y.H.,Moon, J.Y.,Jung, S.S.,Kim, J.O.,Lee, J.C.,Kim, S.Y.,Park, H.S.,Park, M.,Park, D.I.,Lim, D.S.,Jang, K.W.,Lee, J.E. Academic Press 2016 Biochemical and biophysical research communication Vol.479 No.2

Somatic mutation in the tyrosine kinase domain of epidermal growth factor receptor (EGFR) is a decisive factor for the therapeutic response to EGFR tyrosine kinase inhibitors (EGFR-TKIs) in lung adenocarcinoma. The stability of mutant EGFR is maintained by various regulators, including heat shock protein 90 (Hsp90). The C terminus of Hsc70-interacting protein (CHIP) is a Hsp70/Hsp90 co-chaperone and exhibits E3 ubiquitin ligase activity. The high-affinity Hsp90-CHIP complex recognizes and selectively regulates their client proteins. CHIP also works with its own E3 ligase activity independently of Hsp70/Hsp90. Here, we investigated the role of CHIP in regulating EGFR in lung adenocarcinoma and also evaluated the specificity of CHIP's effects on mutant EGFR. In HEK 293T cells transfected with either WT EGFR or EGFR mutants, the overexpression of CHIP selectively decreased the expression of certain EGFR mutants (G719S, L747_E749del A750P and L858R) but not WT EGFR. In a pull-down assay, CHIP selectively interacted with EGFR mutants and simultaneously induced their ubiquitination and proteasomal degradation. The expressions of mutant EGFR in PC9 and H1975 were diminished by CHIP, while the expression of WT EGFR in A549 was nearly not affected. In addition, CHIP overexpression inhibited cell proliferation and xenograft's tumor growth of EGFR mutant cell lines, but not WT EGFR cell lines. EGFR mutant specific ubiquitination by CHIP may provide a crucial regulating mechanism for EGFR in lung adenocarcinoma. Our results suggest that CHIP can be novel therapeutic target for overcoming the EGFR TKI resistance.

서울대학교 保健大學院 博士課程 敎育프로그램의 評價的 硏究

정인숙,박태진,김성옥,배은영,박은옥,김정순,문 용 서울大學校保健大學院 1997 國民保健硏究所硏究論叢 Vol.7 No.2

The doctoral program of school of Public Health in Seoul National University was established in 1976 and about 100 students have graduated or completed the course. Even though there have been several studies on evaluating the educational programs of the school, both master's and doctorate's since early 1980s, this study was carried out to identify needs of students, their performance and perceived achievements, unmet needs and barriers in achieving their educational goals only on doctoral graduates. The study was done by the students of doctoral program taking 'evaluative research in public health services' course as a practice under the direction of the teaching staff. Out of 91 eligibles 51 persons responded to the mail and/or telephone interview, and the results obtained are as followings: 1) The goal of the doctoral program of the Public Health, SNU that was to produce competent public health specialists furnished with full abilities of performong research, education and administration(service)has not been changed since 1976 as was set on the program establishment. 2) The objective of enrollment to the program stated by the subjecs varied considerably; majority of them(47%) for expanding scope of public health sceience, for advanced degree(39%), for contribution to the progress of public health sceience(20%), for better job(20%) and for pure academic interest(2%). 3) The needs of students by area of concern, for instance research, theaching and administration, each classified into several performance is viewed as the proportion accomplished out of needs, which are quantified by 5-point Likert scale, most of the graduates indicated that they have accomplished their needs in 83-92% for research abilities, 75-92% for teaching sbilities and 72-96% for administration abilities, the average performance being the highes in research. 5) The barriers that hindered their performance were the school facilities, such as library(72.5%), deficient funds for research(39.2%), insufficient students' room(33.3%) and deficient laboratory supplies(25.5%). 6) The barrier related to student's characteristics were insufficient study time for part-time student(56.9%), deficient study motivation(45.1%), tendency to take easy curriculum(29.4%) and addmittence of less qualificated students(19.6%). 7) The barrier related to professor's characteristics were deficiency of self-development endeavour(62.7%), lack of professor majoring in some particular fields of public health(39.2%), infrequent collaborative researches/activities with students (17.6%), and absolutely deficient number of professors that are necessary for the program(17.6%). In general, however, the graduates appered to be satisfied in terms of what they have accomplished through the current educational program, which have been revised by the school from time to time to cope with the student's as well as community's demand.

PCR - SSCP 기법을 이용한 소 MC1R 유전자의 다형성 분석 및 한우육 감별

정의룡,김우태,김연수,한상기 한국동물자원과학회 2001 한국축산학회지 Vol.43 No.1

The single-strand conformation polymorphism(SSCP) is a point mutation screening method based on changes in the secondary structure in a defined single-stranded DNA fragment caused by a change in sequence. This study was performed to develop the identification technique of Hanwoo meat using PCR-SSCP marker of MC1R gene associated with the coat colors of cattle. Hanwoo, Holstein and imported cattle breeds(Angus, Hereford and Charolais) were used for the detection of PCR-SSCP genotypes of MC1R gene. The specific primers were used to amplify a 138bp and a 390bp of the bovine E-locus(including point mutation) corresponding to positions 318-455 and 318-707, respectively, in the MC1R gene. For SSCP genotype analysis, the PCR products were denatured at 95℃ for 5min, loaded on a 12-15% nondenaturing polyacrylamide gel and detected by silver staining. When the PCR product of 138bp was separated by gel electrophoresis, two genotypes of E^+/e and e/e were detected only in Hanwoo meat, but E^D/E^D and E^D/e and E^D/E^D genotypes were observed in Holstein meat and Angus breed, respectively. Also, in the SSCP analysis using PCR product of 390bp, Hanwoo meat produced banding pattern of two fragments, while single fragment was observed in Holstein meat and Angus breed. Therefore, breed-specific SSCP markers showing distinct differences between Hanwoo and Holstein and Angus were found by PCR-SSCP analysis. This study shows that the PCR-SSCP technique is relatively simple, fast and a practical tool for determination of MC1R genotypes of cattle breeds when compared with PCR-RFLP method. Therefore, the PCR-SSCP markers of MC1R gene could be used as DNA markers for identification of Hanwoo meat from Holstein and Angus meats.

Heme oxygenase-1 mediates nicotine- and lipopolysaccharide-induced expression of cyclooxygenase-2 and inducible nitric oxide synthase in human periodontal ligament cells

Pi, S.-H.,Jeong, G.-S.,Oh, H.-W.,Kim, Y.-S.,Pae, H.-O.,Chung, H.-T.,Lee, S.-K.,Kim, E.-C. Blackwell Publishing Ltd 2010 Journal of periodontal research Vol.45 No.2

<P><I>Pi S-H, Jeong G-S, Oh H-W, Kim Y-S, Pae H-O, Chung H-T, Lee S-K, Kim E-C. Heme oxygenase-1 mediates nicotine- and lipopolysaccharide-induced expression of cyclooxygenase-2 and inducible nitric oxide synthase in human periodontal ligament cells. J Periodont Res 2010; 45: 177–183. © 2010 John Wiley & Sons A/S</I></P><P>Background and Objective: </P><P>Although heme oxygenase-1 (HO-1) plays a key role in inflammation, its anti-inflammatory effects and mechanism of action in periodontitis are still unknown. This study aimed to identify the effects of HO-1 on the proinflammatory mediators activated by nicotine and lipopolysaccharide (LPS) stimulation in human periodontal ligament (PDL) cells.</P><P>Material and Methods: </P><P>The production of nitric oxide (NO) and prostaglandin E<SUB>2</SUB> (PGE<SUB>2</SUB>) was evaluated using Griess reagent and an enzyme immunoassay, respectively. The expression of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2) and HO-1 proteins was evaluated by Western blot analysis.</P><P>Results: </P><P>Lipopolysaccharide and nicotine synergistically induced the production of NO and PGE<SUB>2</SUB> and increased the protein expression of iNOS, COX-2 and HO-1. Treatment with an HO-1 inhibitor and HO-1 small interfering RNAs blocked the LPS- and nicotine-stimulated NO and PGE<SUB>2</SUB> release as well as the expression of iNOS and COX-2.</P><P>Conclusion: </P><P>Our data suggest that the nicotine- and LPS-induced inflammatory effects on PDL cells may act through a novel mechanism involving the action of HO-1. Thus, HO-1 may provide a potential therapeutic target for the treatment of periodontal disease associated with smoking and dental plaque.</P>

서울대학교 保健大學院 博士課程 敎育프로그램의 評價的 硏究

김정순,김성옥,문용,박은옥,박태진,배은영,정인숙 서울大學校 保健大學院 國民保健硏究所 1997 國民保健硏究所 硏究論叢 Vol.7 No.2

간염 및 소아설사 바이러스질환에 대한 특이진단법 개발

윤재득,지영미,김기순,천두성,정윤석,박정구,황응수,김동수,정용석 국립보건연구원 1999 국립보건원보 Vol.36 No.-

High lipid composition of particulate organic matter in the northern Chukchi Sea, 2011

Kim, B.K.,Lee, J.H.,Yun, M.S.,Joo, H.,Song, H.J.,Yang, E.J.,Chung, K.H.,Kang, S.H.,Lee, S.H. Pergamon Press 2015 Deep-sea research. Part II, Topical studies in oce Vol.120 No.-

We investigated the biochemical compositions (lipids, proteins, and carbohydrates) of particulate organic matter (POM) as a potential food source in the northern Chukchi Sea. We aimed to understand physiological status of phytoplankton, determine important controlling factors, and estimate the energetic contents of POM. The major inorganic nutrients were generally depleted at upper mixed-layer depth (>20m). The average chlorophyll a (chl-a) concentration was 31.9mgm<SUP>-2</SUP> (S.D.=+/-31.3mgm<SUP>-2</SUP>) in this study, significantly higher than that reported previously in the northern Chukchi Sea. Small phytoplankton (0.7-5@?m) accounted for 65.9% of total chl-a concentration. The overall average compositions of lipids, carbohydrates, and proteins were 50% (S.D.=+/-10.7%), 35% (S.D.=+/-11.0%), and 15% (S.D.=+/-11.2%) for POM, respectively. Along with other evidence (e.g., low N:P and protein-carbohydrate ratios), the high lipid and low protein compositions of POM in this study suggests that phytoplankton might have had a nitrogen limitation and/or stationary growth phase in the northern Chukchi Sea during the cruise period, 2011. The overall average calorific content of food material (FM) was 149.2μgL<SUP>-1</SUP> (S.D.=+/-36.5μgL<SUP>-1</SUP>) or 1.0Kcalm<SUP>-3</SUP> (S.D.=+/-0.2Kcalm<SUP>-3</SUP>). The relatively higher calorific contents in the northern Chukchi Sea were due to high lipid contributions and the considerably high calorific content of FM per POC.

세관 양광주 방전에서 플라즈마 확산의 완전 해

김동준,정종문,김정현,황하청,정재윤,조윤희,임현교,구제환,최은하,조광섭,Jin, D.J.,Jeong, J.M.,Kim, J.H.,Hwang, H.C.,Chung, J.Y.,Cho, Y.H.,Lim, H.K.,Koo, J.H.,Choi, E.H.,Cho, G.S. 한국진공학회 2010 Applied Science and Convergence Technology Vol.19 No.1

관경이 수 mm인 세관 램프 내부에서 플라즈마의 확산을 조사하기 위하여 이극성(ambipolar) 확산방정식을 해하였다. 반경 방향의 확산에 의한 유리관 벽에서의 플라즈마 소멸 특성시간은 $\tau_r\;=\;(r_0/2.4)^2/D_a$로 주어진다. 반경 $r_0{\sim}1\;mm$이고 이극성 확산계수 $D_a{\sim}0.01\;m^2/s$ 이면, $\tau_r{\sim}17\;{\mu}s$이다. 이는 램프의 교류전원 구동에서 플라즈마를 유지하기 위한 구동 최소 주파수 ~30 kHz에 해당한다. 고전압이 인가되는 전극부에 발생한 고밀도의 플라즈마가 양광주로 확산되는 특성시간은 $\tau_z{\sim}0.1\;s$이다. 고밀도 플라즈마 경계에서의 시간에 대한 확산속도는 $t{\sim}10^{-6}\;s$일 때 $u_D{\sim}10^2\;m/s$이고, $t{\sim}10^{-3}\;s$이면 그 속도는 $u_D{\sim}1\;m/s$로 느려진다. 따라서 램프 길이 ~1 m에 대하여 전극부에서 생성된 고밀도 플라즈마가 양광주 전체로 확산되는 시간은 수 초가 걸린다. The ambipolar diffusion equation has been solved in a fine-tube lamp of a few mm in diameter. In the diffusion of radial direction, the plasma diffuses and vanishes away at the glass wall by recombination with the characteristic time of plasma loss is given by $\tau_r\;=\;(r_0/2.4)^2/D_a$. With the radius $r_0{\sim}1\;mm$ and the ambipolar diffusion coefficient $D_a{\sim}0.01\;m^2/s$, the vanishing time is calculated $\tau_r{\sim}10\;{\mu}s$ which corresponds to the least value of frequency 30 kHz for the sustaining the plasma in the operation of high voltage AC-power. In the diffusion of longitudinal z-direction, a high density plasma generated at the area of a high voltage electrode, diffuses into the positive column with the characteristic time $\tau_z{\sim}0.1\;s$. The plasma diffusion velocity at the boundary of high density plasma is $u_D{\sim}10^2\;m/s$ at the time $t{\sim}10^{-6}$ s and the diffusion velocity becomes slow as $u_D{\sim}1\;m/s$ at $t{\sim}10^{-3}\;s$. Therefore, for the long lamp of 1 m, it takes about several seconds for the high density plasma at the area of electrode to diffuse through the whole positive column space.

Lipopolysaccharide from <i>Prevotella nigrescens</i> stimulates osteoclastogenesis in cocultures of bone marrow mononuclear cells and primary osteoblasts

Chung, Y.-H.,Chang, E.-J.,Kim, S.-J.,Kim, H.-H.,Kim, H.-M.,Lee, S.-B.,Ko, J. S. Blackwell Publishing Ltd 2006 Journal of periodontal research Vol.41 No.4

<P>Background and Objective: </P><P>Lipopolysaccharide is thought to be a major virulence factor of pathogens associated with periodontal diseases and is believed to stimulate bone resorption <I>in vivo</I>. Although <I>Prevotella nigrescens</I> has been implicated in periodontitis, its role in osteoclastogenesis has not been reported. In this study, we investigated the effects of lipopolysaccharide from <I>P. nigrescens</I> on the formation of osteoclasts and the production of cytokines related to osteoclast differentiation.</P><P>Material and Methods: </P><P>Mouse bone marrow mononuclear cells were cultured in the presence of macrophage colony-stimulating factor (M-CSF) and receptor activator of nuclear factor &kgr;B ligand (RANKL), with or without lipopolysaccharide. Bone marrow mononuclear cells were also cocultured with calvarial osteoblastic cells in the presence or absence of lipopolysaccharide. Osteoclast formation was determined by tartrate-resistant acid phosphatase cytochemistry. The production of osteoprotegerin (OPG), M-CSF, tumor necrosis factor alpha (TNF-&agr;), transforming growth factor-beta (TGF-&bgr;) and prostaglandin E<SUB>2</SUB> (PGE<SUB>2</SUB>) was determined by enzyme-linked immunosorbent assay (ELISA).</P><P>Results: </P><P><I>P. nigrescens</I> lipopolysaccharide inhibited osteoclast differentiation from bone marrow mononuclear cells cultured in the presence of M-CSF and RANKL. However, in the coculture system, <I>P. nigrescens</I> lipopolysaccharide stimulated osteoclastogenesis. Notably, <I>P. nigrescens</I> lipopolysaccharide decreased OPG production but increased TGF-&bgr; secretion. In addition, treatment with <I>P. nigrescens</I> lipopolysaccharide increased PGE<SUB>2</SUB> production during the late stage of the culture period. There was no difference in M-CSF and TNF-&agr; production.</P><P>Conclusion: </P><P>These results demonstrate that <I>P. nigrescens</I> lipopolysaccharide stimulates osteoclastogenesis in the coculture system by decreasing the production of OPG and increasing the production of TGF-&bgr; and PGE<SUB>2</SUB>. Through the mechanisms involving these factors, <I>P. nigrescens</I> lipopolysaccharide may cause alveolar bone resorption in periodontal diseases.</P>