• Title/Summary/Keyword: Endogenous Development

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Reactivation of Silenced WT1 Transgene by Hypomethylating Agents - Implications for in vitro Modeling of Chemoimmunotherapy

  • Kwon, Yong-Rim;Son, Min-Jung;Kim, Hye-Jung;Kim, Yoo-Jin
    • IMMUNE NETWORK
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    • v.12 no.2
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    • pp.58-65
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    • 2012
  • Background: A cell line with transfected Wilms' tumor protein 1 (WT1) is has been used for the preclinical evaluation of novel treatment strategies of WT1 immunotherapy for leukemia due to the lack of appropriate murine leukemia cell line with endogenous WT1. However, silencing of the transgene occurs. Regarding the effects of hypomethylating agents (HMAs) on reactivation of silenced genes, HMAs are considered to be immune enhancers. Methods: We treated murine WT1- transfected C1498 (mWT1-C1498) with increasing doses of decitabine (DAC) and azacitidine (AZA) to analyze their effects on transgene reactivation. Results: DAC and AZA decreased the number of viable cells in a dose- or time-dependent manner. Quantification of WT1 mRNA level was analyzed by real-time polymerase chain reaction after mWT1-C1498 treated with increasing dose of HMA. DAC treatment for 48 h induced 1.4-, 14.6-, and 15.5-fold increment of WT1 mRNA level, compared to untreated sample, at 0.1, 1, and $10{\mu}M$, respectively. Further increment of WT1 expression in the presence of 1 and $10{\mu}M$ DAC was evident at 72 h. AZA treatment also induced up-regulation of mRNA, but not to the same degree as with DAC treatment. The correlation between the incremental increases in WT1 mRNA by DAC was confirmed by Western blot and concomitant down-regulation of WT1 promoter methylation was revealed. Conclusion: The in vitro data show that HMA can induce reactivation of WT1 transgene and that DAC is more effective, at least in mWT1-C1498 cells, which suggests that the combination of DAC and mWT1-C1498 can be used for the development of the experimental model of HMA-combined WT1 immunotherapy targeting leukemia.

A Long Run Classical Model of Price Determination (한국(韓國)의 물가모형(物價模型))

  • Park, Woo-kyu;Kim, Se-jong
    • KDI Journal of Economic Policy
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    • v.14 no.4
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    • pp.3-26
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    • 1992
  • The pupose of this paper is to construct a price determination model of the Korean economy and to find out the propogation mechanism of monetary and fiscal policies. The model is a small-size macroeconometric model consisted of ten core equations : consumption, investment, exports, imports, consumer price index, wage rate, corporate bond rate, potential GNP, capital stock, and GNP identity. The model is a Keynesian model : consumer price index is determined by markup over costs, and wage rate is expressed by Phillipse curve ralation. Two features of the model, however, distinguish this model from other macroeconometric models of the Korean economy. First of all, the estimation of potential GNP and the capital stock is endogenized as suggested by Haque, Lahiri, and Montiel (1990). This allows us to calculate the level of excess demand, which is defined as the difference between the actual GNP and the potential GNP. Second, interest rate, inflation and wages are all estimated as endogenous variables. Moreover, all quantity variables include price variables as important determinants. For instance, interest rate is an important determinant of consumption and investment. Exports and imports are determined by the real effective exchange rate. These two features make the interactions between excess demand and prices the driving forces of this model. In the model, any shock which affects quantity variable(s) affects excess demand, which in turn affects prices. This strong interaction between prices and quantities makes the model look like a classical model over the long run. That is, increases in money supply, government expenditures, and exchange rate (the price of the U.S. dollar in terms of Korean won) all have expansionery effects on the real GNP in the short run, but prices, wage, and interest rate all increase as a result. Over the long run, higher prices have dampenning effects on output. Therefore the level of real GNP turns out to be not much different from the baseline level ; on the other hand, the rates of inflation, wage and interest rate remain at higher levels.

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CK2 phosphorylates AP-2α and increases its transcriptional activity

  • Ren, Kaiqun;Xiang, Shuanglin;He, Fangli;Zhang, Wenfeng;Ding, Xiaofeng;Wu, Yanyang;Yang, Liping;Zhou, Jianlin;Gao, Xiang;Zhang, Jian
    • BMB Reports
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    • v.44 no.7
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    • pp.490-495
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    • 2011
  • Transcription factor AP-$2{\alpha}$ involves in the process of mammalian embryonic development and tumorigenesis. Many studies have shown that AP-$2{\alpha}$ functions in association with other interacting proteins. In a two-hybrid screening, the regulatory subunit ${\beta}$ of protein casein kinase 2 ($CK2{\beta}$) was identified as an interacting protein of AP-$2{\alpha}$; we confirmed this interaction using in-vitro GST pull-down and in-vivo co-immunoprecipitation assays; in an endogenous co-immunoprecipitation experiment, we further found the catalytic subunit ${\alpha}$ of protein casein kinase 2 ($CK2{\alpha}$) also exists in the complex. Phosphorylation analysis revealed that AP-$2{\alpha}$ was phosphorylated by CK2 kinase majorly at the site of Ser429, and such phosphorylation could be blocked by CK2 specific inhibitor 4,5,6,7-tetrabromobenzotriazole (TBB) in a dose-dependent manner. Luciferase assays demonstrated that both $CK2{\alpha}$ and $CK2{\beta}$ enhanced the transcription activity of AP-$2{\alpha}$; moreover, $CK2{\beta}$ increased the stability of AP-$2{\alpha}$. Our data suggest a novel cellular function of CK-2 as a transcriptional co-activator of AP-$2{\alpha}$.

Expression of Transcription Factor FOXC2 in Cervical Cancer and Effects of Silencing on Cervical Cancer Cell Proliferation

  • Zheng, Chun-Hua;Quan, Yuan;Li, Yi-Yang;Deng, Wei-Guo;Shao, Wen-Jing;Fu, Yan
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.4
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    • pp.1589-1595
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    • 2014
  • Objective: Forkhead box C2 (FOXC2) is a member of the winged helix/forkhead box (Fox) family of transcription factors. It has been suggested to regulate tumor vasculature, growth, invasion and metastasis, although it has not been studied in cervical cancer. Here, we analyzed FOXC2 expression in cervical tissues corresponding to different stages of cervical cancer development and examined its correlation with clinicopathological characteristics. In addition, we examined the effects of targeting FOXC2 on the biological behavior of human cervical cancer cells. Methods: The expression of FOXC2 in normal human cervix, CIN I-III and cervical cancer was examined by immunohistochemistry and compared among the three groups and between cervical cancers with different pathological subtypes. Endogenous expression of FOXC2 was transiently knocked down in human Hela and SiHa cervical cells by siRNA, and cell viability and migration were examined by scratch and CCK8 assays, respectively. Results: In normal cervical tissue the frequency of positive staining was 25% (10/40 cases), with a staining intensity (PI) of $0.297{\pm}0.520$, in CIN was 65% (26/40cases), with a PI of $3.00{\pm}3.29$, and in cancer was 91.8% (68/74 cases), with a PI of $5.568 {\pm}3.449$. The frequency was 100% in adenocarcinoma (5/5 cases) and 91.3% in SCCs (63/69 cases). The FOXC2 positive expression rate was 88.5% in patients with cervical SCC stage I and 100% in stage II, showing significant differences compared with normal cervix and CIN. With age, pathologic differentiation degree and tumor size, FOXC2 expression showed no significant variation. On transient transfection of Hela and SiHa cells, FOXC2-siRNA inhibition rates were 76.2% and 75.7%; CCK8 results showed reduced proliferation and relative migration (in Hela cells from $64.5{\pm}3.16$ to $49.5{\pm}9.24$ and in SiHa cells from $60.1{\pm}3.05$ to $44.3{\pm}3.98$) (P < 0.05). Conclusion: FOXC2 gene expression increases with malignancy, especially with blood vessel hyperplasia and invasion degree. Targeted silencing was associated with reduced cell proliferation as well as invasion potential.

Effects of $GA_3$ and ABA Application on After-ripening of Panax quinquefolium Seeds during Stratification ($GA_3$ 및 ABA 사용이 매장처리 중 미국삼 종자의 후숙에 미치는 영향)

  • Guixing Ren;Feng Chen;Haozhe Lian;Jinghui Zhao;Xianzong Gao;Chongming Guo
    • Journal of Ginseng Research
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    • v.20 no.1
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    • pp.83-87
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    • 1996
  • The effects of gibberilin ($GA_3$) on levels of endogenous indole-3-acetic acid (IAA) and zeatin in both fresh and stratified American ginseng (Panax quinquefolium) seeds were investigated. In our first experiment, the fresh seeds were stratified after soaked in 80 ppd $GA_3$ solution for 24 hours. We found that the IAA concentration in embryo increased by 50.7% and 82.1% respectively at the 120th day and the 188th day of stratification, and the zeatin concentration also increased by 3.8% and 51.6% respectively. In our second experiment, we treated the seeds after 134 days stratification with 80 ppm GA3 for 24 hours and then continued to stratify them. We found that the IAA concentration in embryo increased by 32.9% and 17.7% respectively at the 164th day and the 208th day of stratification while zeatin concentration increased by 22.7% and 30.6% respectively In our another experiment, we studied the effects of $GA_3$, abscislc acid (ABA) and GA, plus ABA on germination rate of seeds treated with these plant hormones during stratification. We found that when the stratified seeds whose ratio of embryo had reached 75% were treated with 80 ppm GA3 for 24 hours and then were allowed to be stratified for another 88 days, the weight and length of embryo (p < 0.05), and germination rate (p < 0.01) increased. In contrast, the 25 ppm ABA treated with for 24 hours was found to Inhibit the growth of embryo (p < 0.05) and reduce the germination rate (p < 0.05) . The experiment of combination treatment of $GA_3$ and ABA showed that $GA_3$ could relieve the inhibitory effects of the ABA on the development of the seeds.

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Human Lung Insults due Air Pollutant -A Review for Priority Setting in the Research- (대기오염에 의한 폐장조직 손상 -연구방향의 설정을 위한 논의-)

  • 김건열;백도명
    • Journal of environmental and Sanitary engineering
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    • v.7 no.2
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    • pp.95-110
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    • 1992
  • Much progress has been made in understanding the subcellular events of the human lung injuries after acute exposure to environmental air pollutants. Host of those events represent oxidative damages mediated by reactive oxygen species such as superoxide, hydrogen peroxide, and the hydroxy, free radical. Recently, nitric oxide (NO) was found to be endogenously produced by endothelial cells and cells of the reticulo-endothelial system as endothelialderived relaxation factor (EDRF) which is a vasoactive and neurotransmitter substance. Together with superoxide, NO can form another strong oxidant, peroxonitrite. The relative importance of exogenous sources of $N0/N0_2$ and endogenous production of NO by the EDRF producing enzymes in the oxidative stresses to the heman lung has to be elucidated. The exact events leading to chronic irreversible damage are still yet to be known. From chronic exposure to oxidant gases, progressive epithelial and interstitial damages develop. Type I epithelial cells become thicker and cover a smaller average alveolar surface area while thee II cells proliferate instead. Under acute damages, the extent of loss of the alveolar epithelial cell lining, especially type II cells appears to be a good predictor of the ensuing irreversible damage to alveolar compartment. Interstitial matrix undergo remodeling during chronic exposure with increased collagen fibers and interstitial fibroblasts. However, Inany of these changes can be reversed after cessation of exposure. Among chronic lung injuries, genetic damages and repair responses received particular attention in view of the known increased lung cancer risks from exposure to several air pollutants. Heavy metals from foundry emission, automobile traffics, and total suspended particulate, especially polycystic aromatic hydrocarbons have been positively linked with the development of lung cancer. Asbestos in another air pollutant with known risk of lung cancer and mesothelioma, but asbestos fibers are nonauthentic in most bioassays. Studies using the electron spin resonance spin trapping method show that the presence of iron in asbestos accelerates the production of the hydroxy, radical in vitro. Interactions of these reactive oxygen species with particular cellular components and disruption of cell defense mechanisms still await further studies to elucidate the carcinogenic potential of asbestos fibers of different size and chemical composition. The distribution of inhaled pollutants and the magnitude of their eventual effects on the respiratory tract are determined by pollutant-independent physical factors such as anatomy of the respiratory tract and level and pattern of breathing, as well as by pollutant-specific phyco-chemical factors such as the reactivity, solubility, and diffusivity of the foreign gas in mucus, blood and tissue. Many of these individual factors determining dose can be quantified in vitro. However, mathematical models based on these factors should be validated for its integrity by using data from intact human lungs.

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Associations of Single Nucleotide Polymorphisms in miR-146a, miR-196a, miR-149 and miR-499 with Colorectal Cancer Susceptibility

  • Du, Wei;Ma, Xue-Lei;Zhao, Chong;Liu, Tao;Du, Yu-Liang;Kong, Wei-Qi;Wei, Ben-Ling;Yu, Jia-Yun;Li, Yan-Yan;Huang, Jing-Wen;Li, Zi-Kang;Liu, Lei
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.2
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    • pp.1047-1055
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    • 2014
  • Background: MicroRNAs (miRNAs) are an abundant class of endogenous small non-coding RNAs of 20-25 nucleotides in length that function as negative gene regulators. MiRNAs play roles in most biological processes, as well as diverse human diseases including cancer. Recently, many studies investigated the association between SNPs in miR-146a rs2910164, miR-196a2 rs11614913, miR-149 rs229283, miR-499 rs3746444 and colorectal cancer (CRC), which results have been inconclusive. Methodology/Principal Findings: PubMed, EMBASE, CNKI databases were searched with the last search updated on November 5, 2013. For miR-196a2 rs11614913, a significantly decreased risk of CRC development was observed under three genetic models (dominant model: OR = 0.848, 95%CI: 0.735-0.979, P = 0.025; recessive model: OR = 0.838, 95%CI: 0.721-0.974, P = 0.021; homozygous model: OR = 0.754, 95%CI: 0.627-0.907, P = 0.003). In the subgroup analyses, miR-$196a2^*T$ variant was associated with a significantly decreased susceptibility of CRC (allele model: OR = 0.839, 95%CI: 0.749-0.940, P = 0.000; dominant model: OR = 0.770, 95%CI: 0.653-0.980, P = 0.002; recessive model: OR = 0.802, 95%CI: 0.685-0.939, P = 0.006; homozygous model: OR = 0.695, 95%CI: 0.570-0.847, P = 0.000). As for miR-149 rs2292832, the two genetic models (recessive model: OR = 1.199, 95% CI 1.028-1.398, P = 0.021; heterozygous model: OR = 1.226, 95% CI 1.039-1.447, P = 0.013) demonstrated increased susceptibility to CRC. On subgroup analysis, significantly increased susceptibility of CRC was found in the genetic models (recessive model: OR = 1.180, 95% CI 1.008-1.382, P = 0.040; heterozygous model: OR = 1.202, 95% CI 1.013-1.425, P = 0.013) in the Asian group. Conclusions: These findings supported that the miR-196a2 rs11614913 and miR-149 rs2292832 polymorphisms may contribute to susceptibility to CRC.

Urban Economic Regeneration Strategies of Local Initiative through the Analysis of Regional Strategic Industry Policy Cases (지역전략산업 육성 사례 분석을 통한 지역주도의 도시경제기반형 도시재생 추진 방안)

  • Ryu, Dong-Ju;Kim, Joo-Jin
    • Land and Housing Review
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    • v.7 no.4
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    • pp.239-249
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    • 2016
  • This study aims to find implications for urban economic regeneration strategies of local initiative by analyzing of promoting regional strategic industries, both Seongnam and Goyang in terms of regional industrial policy, institution and specialized service agency. The main results based on the case studies are as follows. First, It is a top priority to formulate the policy direction, such as selecting strategic industries and prepare means for improving it. It should keep reliability and continuity for inducing economic units. Second, It is necessary to consider the effectiveness and diversity of institutions. The institutions to be formalized by municipal ordinance and rules for making the successful implementing system of policies. It is necessary to implement strategic industry policy linked with the central government or public organizations for expanding of a diversity of policies. It is necessary to change in viewpoint on the deregulation and tax break for the private sectors as inducements to achieve the regional economy activation. Third, It is necessary to introduce the specialized service agency to improve an effectiveness and efficiency of institutions and accelerate a network within economic units.

Characterization, detection and identification of transgenic chili pepper harboring coat protein gene that enhances resistance to cucumber mosaic virus

  • Seo, Sang-Gyu;Kim, Ji-Seong;Jeon, Seo-Bum;Shin, Mi-Rae;Kang, Seung-Won;Lee, Gung-Pyo;Hong, Jin-Sung;Harn, Chee-Hark;Ryu, Ki-Hyun;Park, Tae-Sung;Kim, Sun-Hyung
    • Journal of Plant Biotechnology
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    • v.36 no.4
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    • pp.384-391
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    • 2009
  • Previously, two events (H15 and B20) of transgenic pepper (Capsicum annuum L.) that enhanced resistance to Cucumber mosaic virus (CMV) by the introduction of CMV coat protein (CP) gene were constructed. Presently, a single copy number of the CP gene was revealed in H15 and B20 by Southern blot. To predict possible unintended effects due to transgene insertion in an endogenous gene, we carried out sequencing of the 5'-flanking region of the CP gene and a Blastbased search. The results revealed that insertion of the transgene into genes encoding putative proteins may occur in the H15 and B20 transgenic event. Mutiplex polymerase chain reaction (PCR) for simultaneous detection and identification of transgenic pepper was conducted with a set of nine primers. Both transgenic event were differentiated from non-transgenic event by the presence of 267 bp and 430 bp PCR products indicative of CP gene specific primer pairs and primer pairs targeting the CP gene and 35S promoter. H15 and B20 uniquely possessed a 390 bp and 596 bp PCR product, respectively. The presence of a 1115 bp product corresponding to intrinsic pepper actin gene confirmed the use of pepper DNA as the PCR template. The primer set and PCR conditions used presently may allow the accurate and simple identification of CMV resistant transgenic pepper.

Strategy and Task of Government-Funded Research Institution for Post Catch-Up Innovation : Based on case of KRICT(Korea Research Institute of Chemical Technology) (탈추격 혁신을 위한 정부출연연구기관의 노력과 과제: 한국화학연구원을 중심으로)

  • Seong, Ji Eun;Ko, Young Ju
    • Journal of Technology Innovation
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    • v.21 no.2
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    • pp.85-113
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    • 2013
  • This study analyzes the government-funded research institutions as players which derive inner transition based on technology, policy and market environment changes. In this study, the main case is KRICT(Korea Research Institute of Chemical Technology) which tries to make post catch-up innovation in the organization. This institution has attempted to overcome the limitation such as change of technology paradigm and PBS(Project Based System) since 2000's. Responding to the needs of technology convergence and innovation by the paradigm change, KRICT has reformed organization and R&D management system prospectively. And this institution sets the goal like development of natural friendly technologies, small and medium-sized business support, and providing countermeasures of social problems. This case shows possibility of continuous innovation. There are some structural limitation like budget restriction, multiple organization structure, and biased quantitative assessment, but endogenous efforts of government -funded institutions make the new paradigm of post catch-up innovation.

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