• Title/Summary/Keyword: Knock-in

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Aflatoxin B1 Promotes Cell Growth and Invasion in Hepatocellular Carcinoma HepG2 Cells through H19 and E2F1

  • Lv, Jun;Yu, Ya-Qun;Li, Shu-Qun;Luo, Liang;Wang, Qian
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.6
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    • pp.2565-2570
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    • 2014
  • H19 is an imprinted oncofetal gene, and loss of imprinting at the H19 locus results in over-expression of H19 in cancers. Aflatoxin B1(AFB1) is regarded as one of the most dangerous carcinogens. Exposure to AFB1 would most easily increase susceptibility to diseases such as hepatocellular carcinoma(HCC) but any possible relationship between AFB1 and H19 is not clear. In present study, we found that AFB1 could up-regulate the expression of H19 and promote cell growth and invasion by hepatocellular carcinoma HepG2 cells. Knocking down H19 RNA co ld reverse the effects of AFB1 on cell growth and invasion. In addition, AFB1 induced the expression of E2F1 and its knock-down could down-regulate H19 expression and suppress cell growth and invasion in hepatocellular carcinoma HepG2 cells. Furthermore, E2F1 over-expression could up-regulate H19 expression and promote cell growth and invasion, with binding to the H19 promoter being demonstrated by chromatin immunoprecipitation assays (ChIP). In summary, our results suggested that aflatoxin B1could promote cell growth and invasion in hepatocellular carcinoma HepG2 cells through actions on H19 and E2F1.

A Study on Exporting Small & Medium Enterprises Based on Accident Types of Derivatives Transactions: Focus on Exporting Small & Medium-Sized Enterprises with KIKO Currency Option (파생상품의 투자 리스크 요인 분석을 통한 중소수출 기업의 환리스크 관리 방안 - KIKO를 통해 살펴본 국내 중소제조업체를 중심으로 -)

  • Cho, Young-Hun
    • Journal of Arbitration Studies
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    • v.26 no.1
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    • pp.89-105
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    • 2016
  • 2008 began with the American financial crisis which gave way to the liquidity crisis (Fannie Mae and Freddie Mac) situation in which 'the withdrawal of investment initiated from the insufficiency of the U.S. subprime mortgage loan companies', 'the large size loss situation of the financial company (Bear Stearns) due to the American structured bond insufficiency' and the second half opening part national debt mortgage company. Within the American financial crisis was propagated the crisis of international derivatives. Due to this, the withdrawal of foreign investment progressed in the interior of a country with the considerable. By the end of 2007, the exchange rate fluctuation was absorbed in the domestic financial circle on the belief the potentiality of the domestic financial market had been growing drastically through the expansion of the foreign currency debt according to this and it came to the defence but while the exchange rate jumped up to the dollar shortage according to the international crisis, the small and medium companies making the banks and exchange rate-related derivatives contract were going bankrupt due to the derivatives loss. The small and medium factories establish the bank exchange rate-related derivatives has nose (KIKO), pivot (PIVOT), and snowball (Snowball) etc. at that time and the damage which it is the KIKO grasped at 6 end of the months in 2008 caused by reaches to 1 thousand billion 4 thousand hundred million dollars. Small and medium companies in which the dollar which it has to denounce among small and medium companies bearing the KIKO contract in fact with the Knock-In generation city bank exceeds the amount of sales were known to be 68 enterprises among 480 enterprises. This paper departs in this awareness of a problem and tries to look into the risk factor of the derivatives, including nose and study the essential ring risk management plan of small and medium manufacturer.

Knock-down of human MutY homolog (hMYH) decreases phosphorylation of checkpoint kinase 1 (Chk1) induced by hydroxyurea and UV treatment

  • Hahm, Soo-Hyun;Park, Jong-Hwa;Ko, Sung-Il;Lee, You-Ri;Chung, In-Sik;Chung, Ji-Hyung;Kang, Lin-Woo;Han, Ye-Sun
    • BMB Reports
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    • v.44 no.5
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    • pp.352-357
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    • 2011
  • The effect of human MutY homolog (hMYH) on the activation of checkpoint proteins in response to hydroxyurea (HU) and ultraviolet (UV) treatment was investigated in hMYH-disrupted HEK293 cells. hMYH-disrupted cells decreased the phosphorylation of Chk1 upon HU or UV treatment and increased the phosphorylation of Cdk2 and the amount of Cdc25A, but not Cdc25C. In siMYH-transfected cells, the increased rate of phosphorylated Chk1 upon HU or UV treatment was lower than that in siGFP-transfected cells, meaning that hMYH was involved in the activation mechanism of Chk1 upon DNA damage. The phosphorylation of ataxia telangiectasia and Rad3-related protein (ATR) upon HU or UV treatment was decreased in hMYH-disrupted HEK293 and HaCaT cells. Co-immunoprecipitation experiments showed that hMYH was immunoprecipitated by anti-ATR. These results suggest that hMYH may interact with ATR and function as a mediator of Chk1 phosphorylation in response to DNA damage.

Alpha 1,3-Galactosyltransferase Deficiency in Miniature Pigs Increases Non-Gal Xenoantigens

  • Min, Gye-Sik;Park, Jong-Yi
    • Reproductive and Developmental Biology
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    • v.35 no.4
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    • pp.511-518
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    • 2011
  • To avoid hyperacute rejection of xenografts, ${\alpha}1,3$-galactosyltransferase knock-out (GalT KO) pigs have been produced. In this study, we examined whether Sia-containing glycoconjugates are important as an immunogenic non-Gal epitope in the pig liver with disruption of ${\alpha}1,3$-galactosyltransferase gene. The target cells were then used as donor cells for somatic cell nuclear transfer (scNT). A total of 1,800 scNT embryos were transferred to 10 recipients. One recipient developed to term and naturally delivered two piglets. Real-time RT-PCR and glycosyltransferase activity showed that ${\alpha}2,3$-sialyltransferase (${\alpha}2,3ST$) and ${\alpha}2,6$-sialyltransferase (${\alpha}2,6ST$) in the heterozygote GalT KO liver have higher expression levels and activities compared to controls, respectively. According to lectin blotting, sialic acidcontaining glycoconjugate epitopes were also increased due to the decreasing of ${\alpha}$-Gal in heterozygote GalT KO liver, whereas GalNAc-containing glycoconjugate epitopes were decreased in heterozygote GalT KO liver compare to the control. Furthermore, the heterozygote GalT KO liver showed a higher Neu5Gc content than control. Taken together, these finding suggested that the deficiency of GalT gene in pigs resulted in increased production of Neu5Gc-bounded epitopes (H-D antigen) due to increase of ${\alpha}2,6$-sialyltransferase. Thus, this finding suggested that the deletion of CMAH gene to the GalT KO background is expected to further prolong xenograft survival.

p53-mediated HIV-1 Tat Suppression is Likely to be Associated with duble-stranded RNA-dependent Protein Kinase, PKR (p53에 의한 HIV-1 Tat 활성억제와 ds-RNA-dependent Protein Kinase (PKR) 관련 가능성 연구)

  • Kim, Jung-Whan;Byune, Hee-Sun;Bae, Yong-Soo
    • The Journal of Korean Society of Virology
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    • v.29 no.4
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    • pp.235-245
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    • 1999
  • HIV-1 Tat, a strong transactivator, is essential for the HIV-1 replication and AIDS progression. The Tat function is markedly inhibited by human p53 anti-oncogene. However, the detail mechanism has not yet been clearly revealed. In our previous report, we have addressed that p53 is unlikely to interact directly with HIV-1 Tat. In the consecutive experiments, Tat-phosphorylation was found to increase in proportional to the amounts of transfected p53. This work was initiated to identify the signaling factor that is involved in the p53-mediated Tat suppression. Several protein kinases were tested for the phosphorylation of Tat, and we found that PKR is likely to be involved in the p53-mediated Tat suppression. PKR was co-immunoprecipitated by anti-Tat antibody in the Tat-expressing Jurkat cell lysates only when the cells were transfected by p53, indicating that PKR-Tat interaction depends on the p53 activity. The interaction seems to result in PKR-mediated Tat-phosphorylation. Tat function was not blocked by p53 when co-transfected trasiently with antisense-PKR. We have generated PKR-knock out Jurkat cell clone. The PKR defective Jurkat cells didn't show the p53-mediated Tat suppression. These data indicate that p53-mediated Tat suppression is strongly associated with PKR. PKR-mediated Tat phosphorylation experiments are now under investigation by kinase assay and co-immunoprecipitation in the presence or absence of p53.

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Numerical Analysis of Effect of Inhomogeneous Pre-mixture on Pressure Rise Rate in HCCI Engine by Using Multizone Chemical Kinetics (화학반응수치해석을 이용한 HCCI기관의 예혼합기의 성층화성이 연소시의 압력 상승률에 미치는 영향)

  • Lim, Ock-Taeck
    • Transactions of the Korean Society of Mechanical Engineers B
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    • v.34 no.5
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    • pp.449-456
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    • 2010
  • The HCCI engine is a prospective internal combustion engine with which high diesel-like efficiencies and very low NOx and particulate emissions can be achieved. However, several technical issues must be resolved before HCCI engines can be used for different applications. One of the issues concerning the HCCI engine is that the operating range of this engine is limited by the rapid pressure rise caused by the release of excessive heat. This heat release is because of the self-accelerated combustion reaction occurring in the engine and the resulting engine knock in the high-load region. The purpose of this study is to evaluate the role of thermal stratification and fuel stratification in reducing the pressure rise rate in an HCCI engine. The concentrations of NOx and CO in the exhaust gas are also evaluated to confirm combustion completeness and NOx emission. The computation is carried out with the help of a multizone code, by using the information on the detailed chemical kinetics and the effect of thermal and fuel stratification on the onset of ignition and rate of combustion. The engine is fueled with dimethyl ether (DME), which allows heat release to occur in two stages, as opposed to methane, which allows for heat release in a single stage.

Interaction of Microtubule-associated Protein 1B Light Chain(MAP1B-LC1) and p53 Represses Transcriptional Activity of p53

  • Kim, Jung-Woong;Lee, So-Youn;Jeong, Mi-Hee;Jang, Sang-Min;Song, Ki-Hyun;Kim, Chul-Hong;Kim, You-Jin;Choi, Kyung-Hee
    • Animal cells and systems
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    • v.12 no.2
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    • pp.69-75
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    • 2008
  • The tumor suppressor and transcription factor p53 is a key modulator of cellular stress responses, and can trigger apoptosis in many cell types including neurons. In this study, we have shown that Microtubule-associated protein 1B(MAP1B) light chain interacts with tumor suppressor p53. MAP1B is one of the major cytoskeletal proteins in the developing nervous system and essential in forming axons during elongation. We also demonstrate that both p53 and MAP1B-LC1 interact in the nucleus in HEK 293 cells. Indeed, we show that the MAP1B-LC1 negatively regulates p53-dependent transcriptional activity of a reporter containing the p21 promoter. Consequently, MAP1B light chain binds with p53 and their interaction leads to the inhibition of doxorubicin-induced apoptosis in HEK 293 cells. Furthermore, these examinations might be taken into consideration when knock-down of MAP1B-LC1 is used as a cancer therapeutic strategy to enhance p53's apoptotic activity in chemotherapy.

Lean Burn Characteristics in a Heavy Duty Liquid Phase LPG Injection SI Engine (대형 액상분사식 LPG 엔진의 희박연소특성에 관한 연구)

  • O, Seung-Muk;Kim, Chang-Eop;Lee, Jin-Uk;Kim, Chang-Gi;Gang, Geon-Yong;Bae, Chung-Sik
    • 연구논문집
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    • s.33
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    • pp.5-16
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    • 2003
  • Fuel distribution, combustion, and flame propagation characteristics of heavy duty engine with the liquid phase LPG injection(LPLI) were studied in a single cylinder engine. Optically accessible single cylinder engine and laser diagnostics system were built for quantifying fuel concentration by acetone PLIF(planar laser induced fluorescence) measurements. In case of Otto cycle engine with large bore size, the engine knock and thermal stress of exhaust manifold are so critical that lean burn operation is needed to reduce the problems. It is generally known that fuel stratification is one of the key technologies to extend the lean misfire limit. The formation of rich mixture in the spark plug vicinity was achieved by open valve injection. With higher swirl strength(Rs=3.4) and open valve injection, the cloud of fuel followed the flow direction and the radial air/fuel mixing was limited by strong swirl flow. It was expected that axial stratification was maintained with open-valve injection if the radial component of the swirling motion was stronger than the axial components. The axial fuel stratification and concentration were sensitive to fuel injection timing in case of Rs=3.4 while those were relatively independent of the injection timing in case of Rs2.3. Thus, strong swirl flow could promote desirable axial fuel stratification and, in result, may make flame propagation stable in the early stage of combustion.

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Detection of Foreign Antigen-specific $CD4^+Foxp3^+$ Regulatory T Cells by MHC Class II Tetramer and Intracellular CD154 Staining

  • Choi, Jin Young;Eo, Seong Kug
    • IMMUNE NETWORK
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    • v.13 no.6
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    • pp.264-274
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    • 2013
  • The unrestricted population of $CD4^+Foxp3^+$ regulatory T (Treg) cells, which have been known to control the expression of autoimmune diseases and protective immunity to inflammatory reactions, has led to greater appreciation of functional plasticity. Detecting and/or isolating Ag-specific $CD4^+Foxp3^+$ Tregs at the single cell level are required to study their function and plasticity. In this study, we established and compared both MHC class II tetramer and intracellular CD154 staining, in order to detect $CD4^+Foxp3^+$ Treg specific for foreign Ag in acute and chronic infections with lymphocytic choriomeningitis virus (LCMV). Our results revealed that MHC class II tetramer staining showed a lower detection rate of LCMV $GP_{66-77}$-specific $CD4^+$ T cells because most of MHC class II tetramers were unbound and unstable when combined staining was performed with intracellular cytokines. In contrast, intracellular CD154 staining was revealed to be easier and simple for detecting LCMV $GP_{66-77}$-specific $CD4^+$ T cells, compared to MHC class II tetramer staining. Subsequently, we employed intracellular CD154 staining to detect LCMV $GP_{66-77}$-specific $CD4^+Foxp3^+$ Tregs using $Foxp3^{GFP}$ knock-in mouse, and found that LCMV $GP_{66-77}$-specific $CD4^+Foxp3^+$ Tregs and polyclonal $CD4^+Foxp3^+$ Tregs showed differential expansion in mice infected with LCMV Arms or Cl13 at acute (8 and 13 days pi) and chronic phases (35 days pi). Therefore, our results provide insight into the valuable use of intracellular CD154 staining to detect and characterize foreign Ag-specific $CD4^+Foxp3^+$ Treg in various models.

Resveratrol Down-regulates Bmi-1 Expression and Inhibits Breast Cancer Cell Growth In Vitro (유방암세포 성장과 Bmi-1 발현에 대한 레스베라트롤의 억제 효과)

  • Park, Hyun-Joo;Bak, Kwang Je;Ok, Chang Youp;Jang, Hye-Ock;Bae, Moon-Kyoung;Bae, Soo-Kyung
    • KSBB Journal
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    • v.32 no.3
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    • pp.224-232
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    • 2017
  • Resveratrol has been actively investigated as an anticancer drug since it induces cell growth inhibition and apoptosis in many cancer cells. Resveratrol acts through modulation of multiple pathways and genes. In this study, we found resveratrol reduced cell growth and mammosphere formation in MDA-MB-231 triple-negative human breast cancer cells. This suppressive effect of resveratrol is accompanied by a reduction in Bmi-1 gene expression. We also observed that knock-down of Bmi-1 gene by small interfering RNA effectively sensitizes breast cancer cells to resveratrol treatment. Our data demonstrate, for the first time, that resveratrol down-regulates Bmi-1 expression in human breast cancer cells and suggest that specific molecular targeting of Bmi-1 can be combined with a chemotherapeutic strategy to improve the response of breast cancer cells to resveratrol.