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.

New emergence pattern with variant porcine epidemic diarrhea viruses, South Korea, 2012-2015

Chung, H.C.,Lee, J.H.,Nguyen, V.G.,Huynh, T.M.L.,Lee, G.E.,Moon, H.J.,Park, S.J.,Kim, H.K.,Park, B.K. ELSEVIER 2016 VIRUS RESEARCH Vol.226 No.-

Since outbreaks of porcine epidemic diarrhea virus (PEDV) in the United States in 2013, explosive outbreaks of PED in South Korea have infected all age groups of pigs in 2014-2015year. This study analyzed a large collection of the Spike protein coding gene to infer the spatial-temporal diffusion history of PEDV. The studying results suggested that PEDVs in Korea belonged to different genogroups. While classical G1 was continuingly circulating between provinces of Korea, the pandemic G2a were recently introduced from China and USA. By the application of Bayesian phylogeographical analysis, this study demonstrated the spatial-temporal transmission of PEDVs within Korea. Of the recent emerged G2a viruses, J3142 strains showed potential recombination breakpoint (376-2,143nt) of S1 gene between KNU1303_Korea strain_G2a (KJ451046) and 45RWVCF0712_Thailand strain_G2b (KF724935). The pandemic G2a virus was partial neutralized by the antibodies invoked by the G1- based PED vaccine virus.

톱밥 발효사료 급여 및 분말어유의 첨가사료가 돈육의 이화학적 성질에 미치는 영향

이정일,문점동,박구부,박범영,박태선,황보종,이한기,진상근,김영직 한국축산학회 1997 한국축산학회지 Vol.39 No.5

This study was conducted to investigate the influence of pork quality pigs which were fed with diets supplemented with fermented sawdust and sardin powder oil. The pigs were randomly assigned to one of the four treatments: a), control (commercial feed); b), T1 (30% fermented sawdust); c), T2 (10% sardin powder oil) d), T3 (30% fermented sawdust, l0% sardin powder oil and 30% limited amino acid) from 30 kg to 110 kg B. Wt. and slaughtered. The samples were stored at 0±1 C. The physico-chemical properties were determined at the dust of 2, 4, 6, 8 and 15. For the loin the pH's of all the treatments on the 15th day were higher than those of the 2nd day, and that of TI was higher than those of the others. The total moisture contents of all treatments decreased with the storage (P$lt;0.05), and there were no significant differences among treatments on the 15th day. The water soluble proteins of all treatments increased with time and were highest on the 8th day and decreased gianibicanth thereater. T1 had significantly higher water soluble protein than others(P$lt;0.05). The salt soluble proteins of all treatments increased with lure the days(P$lt;0.05). T1 had higher salt soluble protein than others during the storage period(P$lt;0.05). The brittleness, hardness, cohesiveness, gumminess and chewiness of all the treatments were not significantly different among during the period, but elasticity of them increased after from the 2nd day. The brittleness of control was higher than that of the others on the 15th day, and the hardness and chewiness of control were higher than those of the others on the 6th day(P$lt;0.05). Forthebelly the pH's of all the treatments on the 15th day were higher than those of the at 2nd day(P$lt;0.05). T1 showed higher pH than the others during the period(P$lt;0.05). The total moisture contents of control and T1 were significantly decreased with time and there were no significant differences between T2 and T3. T1 showed the highest value. The water soluble proteins of all the treatments decreased with time and was lower on the 15th day(P$lt;0.05). That of control and T1 was significantly higher than that of others on the 2nd day, but there were no significant differences among the treatments on the 15th day. The salt soluble proteins of all treatments increased with time (P$lt;0.05), and they were highest on the 15th day. T1 had significantly higher salt soluble protein than the others on the 8th and 15th days.

A nationwide survey and emission estimates of cyclic and linear siloxanes through sludge from wastewater treatment plants in Korea

Lee, S.,Moon, H.B.,Song, G.J.,Ra, K.,Lee, W.C.,Kannan, K. Elsevier Pub.Co 2014 Science of the Total Environment Vol.497 No.-

Siloxanes are widely used in various industrial applications as well as in personal care products. Despite their widespread use and potential toxic effects, few studies have reported on the occurrence of siloxanes in the environment. In this study, we determined the concentrations of 5 cyclic and 15 linear siloxanes in sludge collected from 40 representative wastewater treatment plants (WWTPs) in Korea. Total concentrations of 20 siloxanes (Σsiloxane) in sludge ranged from 0.05 to 142 (mean: 45.7) μg/g dry weight, similar to the concentrations reported in European countries but higher than those reported in China. The concentrations of siloxanes in sludge from domestic WWTPs were significantly (p<0.01) higher than those from industrial WWTPs, indicating higher consumption of siloxanes in various personal care products (e.g. shampoos and conditioners). The major siloxane compounds found in sludge were decamethylcyclopentasilane (D5), docosamethyldecasiloxane (L10) and dodecamethylcyclohexasilane (D6), which collectively accounted for, on average, 62% of the Σsiloxane concentrations. Non-parametric multidimensional scaling ordination of the profiles of siloxanes indicated the existence of different usage patterns of siloxanes between industrial and household activities. Multiple linear regression analysis of siloxane concentrations and WWTP characteristics suggested that D5, D6 and linear siloxane concentrations in sludge were positively correlated with population served by a WWTP. Environmental emission fluxes of cyclic and linear siloxanes through sludge disposal in Korea were 14,800 and 18,500kg/year, respectively. This is the first report describing occurrence and environmental emission of siloxanes through sludge in Korea.

Inhibitory effects of sulfur compounds on methane oxidation by a methane-oxidizing consortium

Lee, E.H.,Moon, K.E.,Kim, T.G.,Lee, S.D.,Cho, K.S. Society for Bioscience and Bioengineering, Japan ; 2015 Journal of bioscience and bioengineering Vol.120 No.6

Kinetic and enzymatic inhibition experiments were performed to investigate the effects of methanethiol (MT) and hydrogen sulfide (H<SUB>2</SUB>S) on methane oxidation by a methane-oxidizing consortium. In the coexistence of MT and H<SUB>2</SUB>S, the oxidation of methane was delayed until MT and H<SUB>2</SUB>S were completely degraded. MT and H<SUB>2</SUB>S could be degraded, both with and without methane. The kinetic analysis revealed that the methane-oxidizing consortium showed a maximum methane oxidation rate (V<SUB>max</SUB>) of 3.7 mmol g-dry cell weight (DCW)<SUP>-1</SUP> h<SUP>-1</SUP> and a saturation constant (K<SUB>m</SUB>) of 184.1 μM. MT and H<SUB>2</SUB>S show competitive inhibition on methane oxidation, with inhibition values (K<SUB>i</SUB>) of 1504.8 and 359.8 μM, respectively. MT was primary removed by particulate methane monooxygenases (pMMO) of the consortium, while H<SUB>2</SUB>S was degraded by the other microorganisms or enzymes in the consortium. DNA and mRNA transcript levels of the pmoA gene expressions were decreased to ~10<SUP>6</SUP> and 10<SUP>3</SUP>pmoA gene copy number g-DCW<SUP>-1</SUP> after MT and H<SUB>2</SUB>S degradation, respectively; however, both the amount of the DNA and mRNA transcript recovered their initial levels of ~10<SUP>7</SUP> and 10<SUP>5</SUP>pmoA gene copy number g-DCW<SUP>-1</SUP> after methane oxidation, respectively. The gene expression results indicate that the pmoA gene could be rapidly reproducible after methane oxidation. This study provides comprehensive information of kinetic interactions between methane and sulfur compounds.

Performance of electrochemical double layer capacitors using highly porous activated carbons prepared from beer lees

Lee, S.G.,Park, K.H.,Shim, W.G.,balathanigaimani, M.S.,Moon, H. Korean Society of Industrial and Engineering Chemi 2011 Journal of industrial and engineering chemistry Vol.17 No.3

Beer lees precursor is chosen to prepare activated carbons having different physical and chemical properties. The beer lees-based activated carbons (BL-ACs) are characterized by N<SUB>2</SUB> adsorption/desorption isotherms, adsorption energy distributions (AEDs), and X-ray photoelectric spectroscopy (XPS). Furthermore the electrochemical properties of the BL-ACs are assessed using cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS), and constant current charge/discharge method. The maximum specific capacitance (about 188F/g at discharge current of lmA/cm<SUP>2</SUP>) is obtained in 0.1M H<SUB>2</SUB>SO<SUB>4</SUB> electrolyte solution, which is a relatively low electrolyte concentration. The overall results suggest that the BL-ACs are good candidates for EDLC electrode materials.

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.

Characterization of a novel antibody immobilization combining protein G with parylene-H for surface plasmon resonance immunosensors

Shin, E. J.,Lee, W. G.,Moon, S. J. Springer Science + Business Media 2016 Microsystem Technologies Vol.22 No.8

<P>Here we present a method for selectively and efficiently immobilizing antibodies to enhance the detection performance of surface plasmon resonance immune-sensors (SPRIs) for diagnostic applications. To improve the performance of antibody arrays, protein G was used as antibody-selective linkage layer with aldehyde functionalized poly-(para-xylylene) film. To estimate the efficiency of antibody immobilization, immunoglobulin G (IgG) was measured using the anti-IgG immobilized SPRIs. To demonstrate the proof-of-concept validation, the signal detected from the IgG using parylene-H film was compared with that of a combination of parylene-H and protein G in SPRIs. The results showed that the detection of IgG on the immobilized anti-IgG layer using the combination of parylene-H and protein G has a larger change of signal than that of using parylene-H layer. These results also imply that the anti-IgG was densely and efficiently immobilized on the modified surface with the linkage layer in a combination with parylene-H and protein G. Therefore, we believe that this combinatorial approach could selectively immobilize the antibodies, and also be applied for detection and diagnosis of immune diseases in the field of many SPRIs applications.</P>

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.

Porous carbon-coated silica macroparticles as anode materials for lithium ion batteries: Effect of boric acid

Kim, Y.K.,Moon, J.W.,Lee, J.G.,Baek, Y.K.,Hong, S.H. Elsevier Sequoia 2014 Journal of Power Sources Vol.272 No.-

We report carbon-coated porous silica macroparticles (SiO<SUB>2</SUB>Γ) prepared using polymeric templates and subsequent carbonization with sucrose for improved electrochemical energy storage in lithium-ion batteries (LIBs). In addition, boron is introduced to improve the stability of electrochemical cells by pyrolyzing mixtures of sucrose and boric acid (SiO<SUB>2</SUB>Γ + B) under inert atmosphere. The initially large surface area of porous SiO<SUB>2</SUB> (S<SUB>BET</SUB> ~ 658 m<SUP>2</SUP> g<SUP>-1</SUP>) is reduced to 102 m<SUP>2</SUP> g<SUP>-1</SUP> after carbonization and introduction of boric acid. Surface of both SiO<SUB>2</SUB>Γ and SiO<SUB>2</SUB>Γ + B are covered with amorphous carbon. In particular, SiO<SUB>2</SUB>Γ + B particles containing borosilicate (Si-O-B) phase and B-O bondings and Si-C-O bondings are also detected from the X-ray photoelectron spectra. The SiO<SUB>2</SUB>Γ + B macroparticles shows high reversible charge capacity up to 503 mAh g<SUP>-1</SUP> after 103 cycles of Li intercalation/de-intercalation although initial capacity was 200 mAh g<SUP>-1</SUP>. The improved charge capacity of SiO<SUB>2</SUB>Γ + B is attributed to formation of advantageous microstructures induced from boric acid.