• Title/Summary/Keyword: cell library

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DOBI is cleaved by caspases during TRAIL-induced apoptotic cell death

  • Park, Sun-Young;Shin, Jin-Na;Woo, Ha-Na;Piya, Su-Jan;Moon, Ae-Ran;Seo, Young-Woo;Seol, Dai-Wu;Kim, Tae-Hyoung
    • BMB Reports
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    • v.42 no.8
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    • pp.511-515
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    • 2009
  • Downstream of Bid (DOBI) known as Pus10, has been identified as a modulator of TRAIL-induced cell death using RNAi library screening. The crystal structure of DOBI has revealed that it is a crescent-shaped protein containing the pseudouridine synthase catalytic domain and a THUMP-containing domain. Here, we demonstrated that DOBI is expressed in various tissues such as heart and lung, and is also expressed in various tumor cells such as HeLa and A549. Although ectopic expression of DOBI does not promote TRAIL death signaling in HeLa cells, knock-down of DOBI expression using shRNA inhibited TRAIL death signaling. DOBI is cleaved into a 54 kD cleaved DOBI during cell death, and the recombinant DOBI protein can be directly cleaved by caspases-3, or -8 in vitro. Together, these data suggest that the cleaved DOBI may acquire a new function, possibly by cooperating with tBid in the mitochondrial event of cell death caused by TRAIL.

Regulation of signal transducer and activator of transcription 3 activation by dual-specificity phosphatase 3

  • Kim, Ba Reum;Ha, Jain;Kang, Eunjeong;Cho, Sayeon
    • BMB Reports
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    • v.53 no.6
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    • pp.335-340
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    • 2020
  • Since cancer is the leading cause of death worldwide, there is an urgent need to understand the mechanisms underlying cancer progression and the development of cancer inhibitors. Signal transducer and activator of transcription 3 (STAT3) is a major transcription factor that regulates the proliferation and survival of various cancer cells. Here, dual-specificity phosphatase 3 (DUSP3) was identified as a regulator of STAT3 based on an interaction screening performed using the protein tyrosine phosphatase library. DUSP3 interacted with the C-terminal domain of STAT3 and dephosphorylated p-Y705 of STAT3. In vitro dephosphorylation assay revealed that DUSP3 directly dephosphorylated p-STAT3. The suppressive effects of DUSP3 on STAT3 were evaluated by a decreased STAT3-specific promoter activity, which in turn reduced the expression of the downstream target genes of STAT3. In summary, DUSP3 downregulated the transcriptional activity of STAT3 via dephosphorylation at Y705 and also suppressed the migratory activity of cancer cells. This study demonstrated that DUSP3 inhibits interleukin 6 (IL-6)/STAT3 signaling and is expected to regulate cancer development. Novel functions of DUSP3 discovered in IL-6/STAT3 signaling regulation would help expand the understanding of cancer development mechanisms.

Evaluation of Safety of Streptococcus pneumoniae DNA Vaccine in Immunopathological Aspect (폐렴구균 DNA 백신의 면역병리학적 측면에서의 안전성 평가)

  • Lee Jue-Hee;Han Yongmoon
    • YAKHAK HOEJI
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    • v.50 no.1
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    • pp.33-39
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    • 2006
  • We have previously reported the minimum criteria that can be applied to evaluate efficacy and safety of a DNA vaccine with use of Streptococcus pneumoniae DNA vaccine (SPDNA). The SPDNA was formulated by inserting the DNA sequences that are codons specific for the carbohydrate epitope in the capsule of S. penumoniae by phage display peptide library. Administration of the SPDNA into mice induced both humoral and cell-mediated immunities. The induction was protective even in the absence of CD4+ T lymphocyte in mice. Profiles of cytokine and isotyping of antibody displayed tendency of the Th1. In continuation of these studies, we examined if the efficacy of the SPNDA was provoked by the peptide recognized by codons specific for the capsule. Results showed that the peptide vaccine formulae (SPP) induced protective antibody in mice as did the SPDNA. Involvement of the cell-mediated immunity was also determined. Possible side effects of autoimmune diseases such as myositis and C3a production and tumor-formation were undetectable in mice given 7 times of SPDNA vaccination during entire of 92 days. Even after the frequent immunization, immunogenicity of the SPDNA was observed as determined for antibody production, suggesting that there was no immunotolerance provoked. All together, these examining factors would be applied to measurement of a DNA vaccine safety regarding the immunopathological aspect.

Expressed sequence tags (ESTs) analysis of Acanthamoeba healyi

  • Kong, Hyun-Hee;Hwang, Mee-Yeul;Kim, Hyo-Kyung;Chung, Dong-Il
    • Parasites, Hosts and Diseases
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    • v.39 no.2
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    • pp.151-160
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    • 2001
  • Randomly selected 435 clones from Acanthamoeba healyi cDNA library were sequenced and a total of 387 expressed sequence tags (ESTs) had been generated. Based on the results of BLAST search, 130 clones (34.4%) were identified as the genes enconding surface Proteins , enzymes for DNA, energy Production or other metabolism, kinases and phosphatases, protease, proteins for signal transduction, structural and cytoskeletal proteins, cell cycle related proteins, transcription factors, transcription and translational machineries, and transporter proteins. Most of the genes (88.5%) are newly identified in the genus Acanthamoeba. Although 15 clones matched the genes of Acanthamoeba located in the public databases, twelve clones were actin gene which was the most frequently expressed gene in this study. These ESTs of Acanthamoeba would give valuable information to study the organism as a model system for biological investigations such as cytoskeleton or cell movement, signal transduction, transcriptional and translational regulations. These results would also provide clues to elucidate factors for pathogenesis in human granulomatous amoebic encephalitis or keratitis by Acanthamoeba.

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Selective Gene Transfer to Hepatocellular Carcinoma Using Homing Peptide-Grafted Cationic Liposomes

  • Tu, Ying;Kim, Ji-Seon
    • Journal of Microbiology and Biotechnology
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    • v.20 no.4
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    • pp.821-827
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    • 2010
  • Gene delivery that provides targeted delivery of therapeutic genes to the cells of a lesion enhances therapeutic efficacy and reduces toxic side effects. This process is especially important in cancer therapy when it is advantageous to avoid unwanted damage to healthy normal cells. Incorporating cancer-specific ligands that recognize receptors overexpressed on cancer cells can increase selective binding and uptake and, as a result, increase targeted transgene expression. In this study, we investigated whether a peptide capable of homing to hepatocellular carcinoma (HCC) could facilitate targeted gene delivery by cationic liposomes. This homing peptide (HBP) exhibited selective binding to a human hepatocarcinoma cell line, HepG2, at a concentration ranging from 5 to 5,000 nM. When conjugated to a cationic liposome, HBP substantially increased cellular internalization of plasmid DNA to increase the transgene expression in HepG2 cells. In addition, there was no significant enhancement in gene transfer detected for other human cell lines tested, including THLE-3, AD293, and MCF-7 cells. Therefore, we demonstrate that HBP provides targeted gene delivery to HCC by cationic liposomes.

Study of Membrane Potential Analysis According to Applying Doksam-tang to a Human Heart Failure Model (인체심장의 심부전모델에서의 獨蔘湯 투여에 따른 심장막전위 분석에 관한 연구)

  • Jeong, Dae-Yeong;Lee, Boo-Kyun;Hong, Jin-Woo;An, Won-Gun
    • Herbal Formula Science
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    • v.23 no.1
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    • pp.121-131
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    • 2015
  • Objectives : This study was conducted to investigate the membrane potential to apply Doksam-tang to a human heart failure model. Methods : The human heart model was built by Luo et al. CellML model, Priebe et al. CellML model, and a human heart mesh file. Doksam-tang gives channel the half maximal inhibitory concentration(IC 50 ), half maximal effective concentration(EC 50 ) values and compounds concentrations. These data load into the laptop with Ubuntu OS, and build the library with the data. Results : While results of the study with the heart failure model shows abnormal membrane potential from the normal heart model, the study with applying Doksam-tang to heart failure model shows restoring membrane potential that is similar to normal heart model. Conclusions : These results of the testings suggest that a conception of novel technique to investigate the effects of Korean herbal medicine.

CFD CODE DEVELOPMENT FOR THE PREDICTION OF THE SHIP RESISTANCE USING OPEN SOURCE LIBRARIES (소스공개 라이브러리를 활용한 선박 저항계산 CFD 코드 개발)

  • Park, Sun-Ho;Park, Se-Wan;Rhee, Shin-Hyung;Lee, Sang-Bong;Choi, Jung-Eun;Kang, Seon-Hyung
    • Journal of computational fluids engineering
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    • v.17 no.2
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    • pp.21-27
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    • 2012
  • Reynolds-averaged Navier-Stokes equations solver based on a pressure-based cell-centered finite volume method was developed using OpenFOAM libraries, which was an open source and providing computational continuum mechanics libraries. For the reasonable development of the turbulent boundary layer on the bow of the ship, specified library was developed. Grid sensitivities, such as skewness and aspect ratio of a cell, were tested for the solution convergence. Pressure, turbulent kinetic energy, turbulent dissipation rate contours on the ship surface computed by the developed CFD code were compared with those computed by the commercial CFD code, Fluent.

Inhibitory Mechanism of Novel Inhibitors of UDP-N-Acetylglucosamine Enolpyruvyl Transferase from Haemophilus influenzae

  • Jin, Bong-Suk;Han, Seong-Gu;Lee, Won-Kyu;Ryoo, Sung-Weon;Lee, Sang-Jae;Suh, Se-Won;Yu, Yeon-Gyu
    • Journal of Microbiology and Biotechnology
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    • v.19 no.12
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    • pp.1582-1589
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    • 2009
  • Bacterial UDP-N-acetylglucosamine enolpyruvyl transferase (MurA) catalyzes the transfer of enolpyruvate from phosphoenolpyruvate (PEP) to uridine diphospho-N-acetylglucosamine (UNAG), which is the first step of bacterial cell wall synthesis. We identified thimerosal, thiram, and ebselen as effective inhibitors of Haemophilus influenzae MurA by screening a chemical library that consisted of a wide range of bioactive compounds. When MurA was preincubated with these inhibitors, their 50% inhibitory concentrations ($IC_{50}s$) were found to range from 0.1 to $0.7\;{\mu}M$. In particular, thimerosal suppressed the growth of several different Gram-negative bacteria such as Escherichia coli, Pseudomonas aeruginosa, and Salmonella typhimurium at a concentration range of $1-2\;{\mu}g/ml$. These inhibitors covalently modified the cysteine residue near the active site of MurA. This modification changed the open conformation of MurA to a more closed configuration, which may have prevented the necessary conformational change from occurring during the enzyme reaction.

Screening of Gravity Inducible cDNAs in Rice(Oryza sativa L.) Cultured Cell (벼 (Oryza sativa L.)배양세포의 고중력유도성 cDNA의 탐색)

  • ;;Kiyoharu OONO
    • Korean Journal of Plant Tissue Culture
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    • v.21 no.2
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    • pp.111-115
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    • 1994
  • Two different gravity specific cDNA, namely, GSC 13 and GSC 124 with length of 1.34 and 0.67 kilobase pairs, and transcripts of 2.0 and 1.9 kilobase pairs, respectively. were isolated by differential screening and northern hybridization of the total RNA isolated from treated and untreated cultured cells showed that maximum levels of trannscripts were achieved after 4 h of gravity stress at 450, 000 x g for both, GSC 13 and GSC 124, suggesting that these mRNA could be expressed and translated into polyeptites related to the cell to extream gravity stress.

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Cellular Mechanisms of a New Pyrazinone Compound that Induces Apoptosis in SKOV-3 Cells

  • Wang, Guan;Jiang, Meng-Ying;Meng, Ying;Song, Hong-Rui;Shi, Wei
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.2
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    • pp.797-802
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    • 2014
  • We screened a small molecular library that was designed and independently synthesized in vitro and found a new drug (MY-03-01) that is active against ovarian cancer. We established that MY-03-01 effectively inhibited SKOV-3 cell survival in a dose-dependent manner, based on cell viability rates, and that it not only induced SKOV-3 apoptosis by itself, but also did so synergistically with paclitaxel. Secondly, when MY-03-01 was applied at $40{\mu}M$, its hemolytic activity was less than 10%, compared with the control, and there was almost no damage to nor mal cells at this concentration. In addition, we used DAPI staining and flow cytometry to show that MY-03-01 could significantly induce apoptosis of SKOV-3 cells. Finally, we found that MY-03-01 likely induced SKOV-3 apoptosis by activating caspase3 and caspase9 through the mitochondrial pathway.