Gα₁₂ gep oncogene deregulation of p53-responsive microRNAs promotes epithelial–mesenchymal transition of hepatocellular carcinoma

Yang, Y M,Lee, W H,Lee, C G,An, J,Kim, E-S,Kim, S H,Lee, S-K,Lee, C H,Dhanasekaran, D N,Moon, A,Hwang, S,Lee, S J,Park, J-W,Kim, K M,Kim, S G Macmillan Publishers Limited 2015 Oncogene Vol.34 No.22

Hepatocellular carcinoma (HCC) has a poor prognosis owing to aggressive phenotype. Gα<SUB>12</SUB> gep oncogene product couples to G-protein-coupled receptors, whose ligand levels are frequently increased in tumor microenvironments. Here, we report Gα<SUB>12</SUB> overexpression in human HCC and the resultant induction of zinc-finger E-box-binding homeobox 1 (ZEB1) as mediated by microRNA deregulation. Gα<SUB>12</SUB> expression was higher in HCC than surrounding non-tumorous tissue. Transfection of Huh7 cell with an activated mutant of Gα<SUB>12</SUB> (Gα<SUB>12</SUB>QL) deregulated microRNA (miRNA or miR)-200b/a/429, -194-2/192 and -194-1/215 clusters in the miRNome. cDNA microarray analyses disclosed the targets affected by Gα<SUB>12</SUB> gene knockout. An integrative network of miRNAs and mRNA changes enabled us to predict ZEB1 as a key molecule governed by Gα<SUB>12</SUB>. Decreases of miR-200a/b, -192 and -215 by Gα<SUB>12</SUB> caused ZEB1 induction. The ability of Gα<SUB>12</SUB> to decrease p53 levels, as a result of activating protein-1 (AP-1)/c-Jun-mediated mouse double minute 2 homolog induction, contributed to transcriptional deregulation of the miRNAs. Gα<SUB>12</SUB>QL induced ZEB1 and other epithelial–mesenchymal transition markers with fibroblastoid phenotype change. Consistently, transfection with miR-200b, -192 or -215 mimic prevented the ability of Gα<SUB>12</SUB>QL to increase tumor cell migration/invasion. In xenograft studies, sustained knockdown of Gα<SUB>12</SUB> decreased the overall growth rate and average volume of tumors derived from SK-Hep1 cell (mesenchymal-typed). In HCC patients, miR-192, -215 and/or -200a were deregulated with microvascular invasion or growth advantage. In the HCC samples with higher Gα<SUB>12</SUB> level, a correlation existed in the comparison of relative changes of Gα<SUB>12</SUB> and ZEB1. In conclusion, Gα<SUB>12</SUB> overexpressed in HCC causes ZEB1 induction by deregulating p53-responsive miRNAs, which may facilitate epithelial–mesenchymal transition and growth of liver tumor. These findings highlight the significance of Gα<SUB>12</SUB> upregulation in liver tumor progression, implicating Gα<SUB>12</SUB> as an attractive therapeutic target.

The gep oncogenes, Gα₁₂ and Gα₁₃, upregulate the transforming growth factor-β1 gene

Lee, S J,Yang, J W,Cho, I J,Kim, W D,Cho, M K,Lee, C H,Kim, S G Macmillan Publishers Limited 2009 Oncogene Vol.28 No.9

Transforming growth factor-β1 (TGFβ1) plays a role in neoplastic transformation and transdifferentiation. Gα<SUB>12</SUB> and Gα<SUB>13</SUB>, referred to as the gep oncogenes, stimulate mitogenic pathways. Nonetheless, no information is available regarding their roles in the regulation of the TGFβ1 gene and the molecules linking them to gene transcription. Knockdown or knockout experiments using murine embryonic fibroblasts and hepatic stellate cells indicated that a Gα<SUB>12</SUB> and Gα<SUB>13</SUB> deficiency reduced constitutive, auto-stimulatory or thrombin-inducible TGFβ1 gene expression. In contrast, transfection of activated mutants of Gα<SUB>12</SUB> and Gα<SUB>13</SUB> enabled the knockout cells to promote TGFβ1 induction. A promoter deletion analysis suggested that activating protein 1 (AP-1) plays a role in TGFβ1 gene transactivation, which was corroborated by the observation that a deficiency of the G-proteins decreased the AP-1 activity, whereas their activation enhanced it. Moreover, mutation of the AP-1-binding site abrogated the ability of Gα<SUB>12</SUB> and Gα<SUB>13</SUB> to induce the TGFβ1 gene. Transfection of a dominant-negative mutant of Rho or Rac, but not Cdc42, prevented gene transactivation and decreased AP-1 activity downstream of Gα<SUB>12</SUB> and Gα<SUB>13</SUB>. In summary, Gα<SUB>12</SUB> and Gα<SUB>13</SUB> regulate the expression of the TGFβ1 gene through an increase in Rho/Rac-dependent AP-1 activity, implying that the G-protein-coupled receptor (GPCR)-Gα<SUB>12</SUB> pathway is involved in the TGFβ1-mediated transdifferentiation process.Oncogene (2009) 28, 1230–1240; doi:10.1038/onc.2008.488; published online 19 January 2009

Full-length genomic analysis of porcine G9P[23] and G9P[7] rotavirus strains isolated from pigs with diarrhea in South Korea

Kim, H.H.,Matthijnssens, J.,Kim, H.J.,Kwon, H.J.,Park, J.G.,Son, K.Y.,Ryu, E.H.,Kim, D.S.,Lee, W.S.,Kang, M.I.,Yang, D.K.,Hyun, B.H.,Park, S.I.,Park, S.J.,Cho, K.O. Elsevier Science 2012 INFECTION GENETICS AND EVOLUTION Vol.12 No.7

Group A rotaviruses (RVAs) are agents causing severe gastroenteritis in infants and young animals. G9 RVA strains are believed to have originated from pigs. However, this genotype has emerged as the fifth major human RVA genotype worldwide. To better understand the relationship between human and porcine RVA strains, complete RVA genome data are needed. For human RVA strains, the number of complete genome data have grown exponentially. However, there is still a lack of complete genome data on porcine RVA strains. Recently, G9 RVA strains have been identified as the third most important genotype in diarrheic pigs in South Korea in combinations with P[7] and P[23]. This study is the first report on complete genome analyses of 1 G9P[7] and 3 G9P[23] porcine RVA strains, resulting in the following genotype constellation: G9-P[7]/P[23]-I5-R1-C1-M1-A8-N1-T1-E1-H1. By comparisons of these genotype constellations, it was revealed that the Korean G9P[7] and G9P[23] RVA strains possessed a typical porcine RVA backbone, similar to other known porcine RVA strains. However, detailed phylogenetic analyses revealed the presence of intra-genotype reassortments among porcine RVA strains in South Korea. Thus, our data provide genetic information of G9 RVA strains increasingly detected in both humans and pigs, and will help to establish the role of pigs as a source or reservoir for novel human RVA strains.

MINERAL NUTRITION OF GRAZING SHEEP IN NORTHERN CHINA I. MACRO-MINERALS IN PASTURE, FEED SUPPLEMENTS AND SHEEP

Masters, D.G.,Purser, D.B.,Yu, S.X.,Wang, Z.S.,Yang, R.Z.,Liu, N.,Lu, D.X.,Wu, L.H.,Ren, J.K.,Li, G.H. Asian Australasian Association of Animal Productio 1993 Animal Bioscience Vol.6 No.1

This study determined the macro-mineral levels in plants and sheep, at different times during the year, at three farms in northern China. Samples of plants, animal tissues and faeces were collected at 5 to 8 times during the year from each site. They were analysed for calcium, sodium, phosphorus, magnesium and potassium. Sodium concentrations in plants were below those recommended for optimum animal production at all sites for all or part of the year (0.01-1.66 g/kg DM). Low concentrations of sodium in faeces were measured and signs of sodium deficiency (soil ingestion) were observed on one farm. There were seasonal trends in other mineral levels in plants and animals. Plants were lowest in potassium (2.3-13.4 g/kg DM), magnesium (1.28-4.82 g/kg DM) and phosphorus (0.24-1.62 g/kg DM) in winter and spring. However, high levels of these elements were supplied in the feed supplements used at this time of the year. During the periods of rapid pasture growth, in summer and autumn, supplements of feed and salt are often not provide even though pasture concentrations of phosphorus and sodium are low. It may be at these times that sheep will be most susceptible to deficiencies of these elements.

한국 재래 닭 부화 후 고환 발달에 관한 연구

장병귀,태현진,최철환,박영재,박병용,박상열,강형섭,김남수,이영훈,양홍현,안동춘,김인식,Jang, B.G.,Tae, H.J.,Choi, C.H.,Park, Y.J.,Park, B.Y.,Park, S.Y.,Kang, H.S.,Kim, N.S.,Lee, Y.H.,Yang, H.H.,Ahn, D.C.,Kim, I.S. 한국가금학회 2006 韓國家禽學會誌 Vol.33 No.3

이 연구는 한국 재래 닭에서 부화 후 고환 발달 과정을 명확하게 이해가기 위하여 부화 후 1, 2, 4, 6, 8, 10, 12, 14, 16, 18, 21, 28, 32, 44, 52 및 64주령(n=13마리/일령)의 한국 재래닭을 이용하여 수행하였다. 한국 재래 닭의 고환은 2.5 % glutaraldehyde를 이용하여 전신 관류 고정하고 조직 처리 과정을 거쳐 Epon-araldite에 포매하였다. 초박절편기를 사용하여 $1{\mu}m$로 절편한 다음 methylene blue로 염색하여 일반적인 조직의 변화상과 형태 계측을 일반적인 방법에 따라서 정자 생성을 측정하여 다음과 같은 결과를 얻었다. 부화 후 1주령의 고환의 평균 무게는 0.015 g이었고 점진적으로 증가하여 21주령에는 3.93 g이고 21주령부터 64주령까지는 변화가 없었다. 곱슬정세관의 용적 치밀도는 1주령에 32.6%이었으나 점차적으로 증가하여 64주령에서는 92.89이었다. 1주령의 한국 재래 닭 고환 간질 조직은 고환 실질의 67.4%를 나타내었고 이러한 비율은 성장하는 동안에 점차적으로 감소하여 64주령에 7.11%를 나타내었다. 고환내 총 정자 생성은 18주령부터 28주령까지는 유의성있게 증가하였고 고환 1g당 정자 생성은 $18\sim28$주령까지는 유의성있게 증가하였고 $28\sim52$주령까지는 변화가 없었으나 64주령에 유의성 있게 감소하였다. 곱슬정세관의 평균 직경은 $1\sim21$주령까지 주령에 따라 점진적으로 증가하였고 곱슬정세관의 길이는 1주령에 0.34 m이었고 성장하면서 유의성 있게 증가하여 64주령에서는 72.2 m이었다. 곱슬정세관내 생식세포의 발달 단계는 다음과 같이 분류할 수 있다. 1) 정조세포($1\sim8$주령), 2) 정조세포, 정모세포($10\sim12$주령), 3) 정조세포, 정모세포, 원형의 정자세포($14\sim16$주령), 4) 정조세포, 정모세포, 정자세포 및 정자($18\sim64$주령). 이러한 결과를 종합하여 보면 한국 재래 닭에서 부화 후부터 성숙시기까지 고환 발달의 양상은 신생시기-성 성숙 이전기($1\sim12$주령), 성 성숙시기($14\sim18$주령) 및 성숙시기$(21\sim64)$로 나뉜다. Changes in the chicken testis from hatching to adulthood were studied in Korean native chickens of 1, 2, 4, 6, 8, 10, 12, 14, 16, 18, 21, 24, 28, 32, 44, 52 and 64 weeks (n=13 chickens per group) of age. The present study was to investigate in more detail the post-hatching development of testis in Korean native chickens. Testes of chickens were fixed by whole body perfusion using a fixative containing 2.5% glutaraldehyde in cacodylate buffer, processed and embedded in Epon-araldite. Using $1{\mu}m$ sections stained with methylene blue-azure II, qualitative and quantitative(stereological) morphological studies were performed. Sperm production was measured by routine technique. The average volume of a testis of 1 week old Korean native chickens was determined as 0.015 g and the parameter increased linearly from 1 week to 21 weeks days (28.9 g), and did not change from 21 weeks to 64 weeks. The volume density of the seminiferous tubules increased with age from 32.6% at week 1 to 92.89% at week 64. The volume density of the interstitium represents 67.4% of the testicular parenchyma at week 1. This proportion progressively diminished during development to reach a value of 7.11% at week 64. Total sperm production per testis increased significantly from 18 weeks to 28 weeks and remained unchanged. Sperm production per 1 g testis increased significantly from 18 weeks to 28 weeks, did not change significantly from 28 weeks to 52 weeks, and declined significantly at 64 weeks of age. The average diameter of the seminiferous tubules gradually increased with age from 1 week $(42.4{\mu}m)$ to 21 weeks $(412.8{\mu}m)$. The length of the seminiferous tubules was 0.34 m at 1 week, increased significantly in subsequent age groups and reached 72.2 m by weeks 64. The stage of germ cell development in seminiferous tubules was classified as 1) spermatogonia $(1\sim8\;weeks)$, 2) spermatogonia and spermatocytes $(10\sim12\;weeks)$, 3) spermatogonia, spermatocytes and round spermatids $(14\sim16\;weeks)$, and 4) speramatogonia, spermatocytes, spermatids and spermatozoa $(18\sim64\;weeks)$. These results clarified the pattern of changes in the testicular development in Korean native chickens from hatching to adulthood as 1) neonatal-prepubertal $(1\sim12\;weeks)$, 2) puberty$(14\sim18\;weeks)$, and adult$(21\sim64\;weeks)$.

Power supply system for KSTAR neutral beam injector

Cho, W.,Bae, Y.S.,Han, W.S.,Jeong, J.H.,Kim, J.S.,Park, H.T.,Yang, H.L.,Oh, Y.K.,Kwak, J.G. Elsevier 2015 Fusion engineering and design Vol.96 No.-

<P><B>Abstract</B></P> <P>The power supply system in KSTAR neutral beam injector consists of low voltage and high current DC power supplies for plasma generator of ion source and high voltage and high current DC power supply for accelerator grid system. The arc discharge is initiated by an arc power supply supplying the arc voltage between the chamber wall and 12 filaments which are heated by individual filament power supply. The negative output of arc power supply is common to each positive output of 12 filament power supplies. To interrupt the arc discharging for the fault condition of the arc current unbalance, DCCT current monitor is placed at the positive output cable of the filament power supply. The plasma grid (G1) power supply has the maximum capability of 120kV/70A which consists of low voltage regulator with IGBT-switched chopper array system for the voltage control in unit of 600V and the high voltage rectified transformers to supply DC voltage of 20kV, 30kV, and 50kV. The output voltage of the G1 power supply is also connected to the input of the voltage divider system which supplies the gradient voltage to the gradient grid (G2) in the range of 80–90% of G1 voltage by changing tap of winding resistors in unit of 1%. The charged G1 voltage is turned on and off by the high voltage switch (HVS) system consisting of MOSFET fast semiconductor switches which can immediately be opened less than 1μs when the ion source grid breakdown occurs. The decelerating grid (G3) power supply is inverter system using capacitor-charge power supply to supply maximum −5kV/5A. The important component in power supply system is the surge absorber near the ion source to limit the arc energy deposited to accelerator grid. This paper presents configuration and features of power supply system, main controller, and interlock system of KSTAR NBI.</P> <P><B>Highlights</B></P> <P> <UL> <LI> The power supply system in KSTAR NBI consists of DC power supplies for ion source. </LI> <LI> For operation NBI, DC High Voltage based on the low voltage transformer with chopper. </LI> <LI> The surge absorber near the ion source limit the energy deposited to accelerator grid. </LI> </UL> </P>

Structural and Functional Analysis of a β₂-Adrenergic Receptor Complex with GRK5

Komolov, Konstantin E.,Du, Yang,Duc, Nguyen Minh,Betz, Robin M.,Rodrigues, Joã,o P.G.L.M.,Leib, Ryan D.,Patra, Dhabaleswar,Skiniotis, Georgios,Adams, Christopher M.,Dror, Ron O.,Chung, Ka Young Cell Press 2017 Cell Vol. No.

<P><B>Summary</B></P> <P>The phosphorylation of agonist-occupied G-protein-coupled receptors (GPCRs) by GPCR kinases (GRKs) functions to turn off G-protein signaling and turn on arrestin-mediated signaling. While a structural understanding of GPCR/G-protein and GPCR/arrestin complexes has emerged in recent years, the molecular architecture of a GPCR/GRK complex remains poorly defined. We used a comprehensive integrated approach of cross-linking, hydrogen-deuterium exchange mass spectrometry (MS), electron microscopy, mutagenesis, molecular dynamics simulations, and computational docking to analyze GRK5 interaction with the β<SUB>2</SUB>-adrenergic receptor (β<SUB>2</SUB>AR). These studies revealed a dynamic mechanism of complex formation that involves large conformational changes in the GRK5 RH/catalytic domain interface upon receptor binding. These changes facilitate contacts between intracellular loops 2 and 3 and the C terminus of the β<SUB>2</SUB>AR with the GRK5 RH bundle subdomain, membrane-binding surface, and kinase catalytic cleft, respectively. These studies significantly contribute to our understanding of the mechanism by which GRKs regulate the function of activated GPCRs.</P> <P><B>PaperClip</B></P> <P>Display Omitted</P> <P><B>Highlights</B></P> <P> <UL> <LI> GRK5-β<SUB>2</SUB>AR binding is enhanced by receptor and kinase ligands and acidic lipids </LI> <LI> GRK5 binding to the β<SUB>2</SUB>AR involves a multi-site interaction </LI> <LI> Receptor binding triggers substantial conformational changes in GRK5 </LI> <LI> RH/catalytic domain separation in GRK5 is essential for receptor phosphorylation </LI> </UL> </P> <P><B>Graphical Abstract</B></P> <P>[DISPLAY OMISSION]</P>

The quantitative assessment of MHC II on thymic epithelium: implications in cortical thymocyte development

Yang, Soo Jung,Ahn, Sejin,Park, Chan Sik,Holmes, Kevin L,Westrup, Jenifer,Chang, Cheong Hee,Kim, Moon G Oxford University Press 2006 International immunology Vol.18 No.5

<P>The dynamics of MHC II expression in various thymic stromal compartments was investigated. By including MHC II in flow cytometry in addition to the cortical CDR1, medullary UEA-1 and pan-epithelial G8.8 markers, thymic stromal compartments were subdivided into at least six different populations. The total level of surface and cytoplasmic MHC II from fresh cortical thymic epithelial cells (cTECs) of normal mouse was as high as MHC II levels in medullary thymic epithelial cells (mTECs). MHC II levels as well as the percentages and cycling status of thymic epithelial cell populations expressing MHC II were not static during post-natal development, suggesting quantitative flexibility in presenting signals to the developing thymocytes. Although there was no evidence found for regulation of surface MHC II levels by TCR or by IFN-γ, the absence of class II transactivator reduced both the level of MHC II expression and the number of MHC II<SUP>+</SUP> cells. Surprisingly, MHC II molecules were found to form distinct focal aggregates on the surface of cTEC but not mTEC using high-resolution analysis by confocal microscopy. Moreover, these aggregates were formed independent of TCR or TCR-bearing cells in the thymus. These aggregates could potentially generate a functional unit containing a much higher local MHC II concentration to yield a higher avidity interaction. We discuss possible mechanisms for positive selection by weak interactions in the presence of such preformed MHC II aggregate units in cTEC.</P>

Mouse수정란의 초급속동결에 있어서 Vitrification Solution 개발에 관한 연구 : Ⅱ. Vitrification Solution 내의 비투과성 물질(Ficoll, sucrose)과 평형시간이 초급속동결· 융해후 mouse morulae의 생존율에 미치는 영향

고경래,김중계,강민수,장덕지,양병철 濟州大學校 農科大學 動物科學硏究所 1992 動物科學論叢 Vol.7 No.1

This study was carried out to study effects of the addition level of acetamide and non-permea-ble cryoprotectants(Ficoll, sucrose) in VS(20% glycerol + 10% ethyleneglycol) and equilibration time on the survival of vitrified mouse morulae. The results are summarized as follows: 1. When 10, 15 and 20% of acetamide were added to the new vitrification solution(20G 10E), FDA-scores of embryos were 4.4(control), 4.4(10%), 3.6(15, 20%), respectively. The addition of acetamide did not affect the survival of forzen-thawed morulae. 2. The survival between 5 min(3.5) and 10 min(4.6), 10 min(4.6) and 20 min(3.2) of equili-bration in 10% sucmse, and 20 min(3.2) and 5 min(4.0), or 10 min(4.3) in 20% sucrose was sig-nificantly different(P<0.05). The highest survival(4.6) was obtained in mouse morulae equilibrat-ed in VS(2OG10E) containing 10% sucrose for 10 minutes. 3. FDA-score of morulae frozen in the new vitrification solution containing 0, 10, 20 and 30% Ficoll was 4.5, 4.2, 4.4 and 4.6, respectively and had no significant effect among concentrations of Ficoll. The development rate after culture(24h) was 89%(20% Ficoll) and 93% (30% Ficoll), respectively.

자동차 항법용 자이로 센서 시스템의 특성분석 및 Hybrid항법 알고리즘 개발

김상겸(S.G.Kim),최정훈(J.H.Choi),김정하(J.H.Kim),전진홍(J.H.Chun),양승규(S.G.Yang) 한국자동차공학회 1997 한국자동차공학회 춘 추계 학술대회 논문집 Vol.1997 No.6_1

Generally. G.P.S ( Global Positioning System ) is using for the car navigation system but it has some restrictions such as the discontinuity of earth satellites and SA ( Selective Availability ). Recently, the hybrid navigation system combining with G.P.S and Dead-Reckoning are much attractive for improving the accuracy of a vehicle positioning. G.P.S called satellite navigation system, can measure its position by using satellites. Dead-Reckoning is the self-contained navigation system, using wheel sensor for the vehicle velocity and gyro sensor for the vehicle angular velocity. Some algorithm could be generated for finding the vehicle position and orientation. In this paper, a Hybrid algorithm is developed and analyzed the characteristic of a Piezoelectric Vibrating Gyroscope & GPS.<br/>