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29-kDa FN-f inhibited autophagy through modulating localization of HMGB1 in human articular chondrocytes

  • Hwang, Hyun Sook;Choi, Min Ha;Kim, Hyun Ah
    • BMB Reports
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    • v.51 no.10
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    • pp.508-513
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    • 2018
  • Fibronectin fragments found in the synovial fluid of patients with osteoarthritis (OA) induce the catabolic responses in cartilage. Nuclear high-mobility group protein Box 1 (HMGB1), a damage-associated molecular pattern, is responsible for the regulation of signaling pathways related to cell death and survival in response to various stimuli. In this study, we investigated whether changes induced by 29-kDa amino-terminal fibronectin fragment (29-kDa FN-f) in HMGB1 expression influences the pathogenesis of OA via an HMGB1-modulated autophagy signaling pathway. Human articular chondrocytes were enzymatically isolated from articular cartilage. The level of mRNA was measured by quantitative real-time PCR. The expression of proteins was examined by western blot analysis, immnunofluorescence assay, and enzyme-linked immunosorbent assay. Interaction of proteins was evaluated by immunoprecipitation. The HMGB1 level was significantly lower in human OA cartilage than in normal cartilage. Although 29-kDa FN-f significantly reduced the HMGB1 expression at the mRNA and protein levels 6 h after treatment, the cytoplasmic level of HMGB1 was increased in chondrocytes treated with 29-kDa FN-f, which significantly inhibited the interaction of HMGB1 with Beclin-1, increased the interaction of Bcl-2 with Beclin-1, and decreased the levels of Beclin-1 and phosphorylated Bcl-2. In addition, the level of microtubule-associated protein 1 light chain 3-II, an autophagy marker, was down-regulated in chondrocytes treated with 29-kDa FN-f, whereas the effect was antagonized by mTOR knockdown. Furthermore, prolonged treatment with 29-kDa FN-f significantly increased the release of HMGB1 into the culture medium. These results demonstrated that 29-kDa FN-f inhibits chondrocyte autophagy by modulating the HMGB1 signaling pathway.

Human Endogenous Retrovirus K (HERV-K) can drive gene expression as a promoter in Caenorhabditis elegans

  • Durnaoglu, Serpen;Kim, Heui-Soo;Ahnn, Joohong;Lee, Sun-Kyung
    • BMB Reports
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    • v.53 no.10
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    • pp.521-526
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    • 2020
  • Endogenous retroviruses (ERVs) are retrotransposons present in various metazoan genomes and have been implicated in metazoan evolution as well as in nematodes and humans. The long terminal repeat (LTR) retrotransposons contain several regulatory sequences including promoters and enhancers that regulate endogenous gene expression and thereby control organismal development and response to environmental change. ERVs including the LTR retrotransposons constitute 8% of the human genome and less than 0.6% of the Caenorhabditis elegans (C. elegans) genome, a nematode genetic model system. To investigate the evolutionarily conserved mechanism behind the transcriptional activity of retrotransposons, we generated a transgenic worm model driving green fluorescent protein (GFP) expression using Human endogenous retroviruses (HERV)-K LTR as a promoter. The promoter activity of HERV-K LTR was robust and fluorescence was observed in various tissues throughout the developmental process. Interestingly, persistent GFP expression was specifically detected in the adult vulva muscle. Using deletion constructs, we found that the region from positions 675 to 868 containing the TATA box was necessary for promoter activity driving gene expression in the vulva. Interestingly, we found that the promoter activity of the LTR was dependent on che-1 transcription factor, a sensory neuron driver, and lin-15b, a negative regulator of RNAi and germline gene expression. These results suggest evolutionary conservation of the LTR retrotransposon activity in transcriptional regulation as well as the possibility of che-1 function in non-neuronal tissues.

A Basic Study on Vehicle Load Analyzing System for Embedded Road (임베디드 도로를 위한 차량하중 분석시스템 기초연구)

  • Jo, Byung-Wan;Yoon, Kwang-Won;Park, Jung-Hoon;Kim, Heoun
    • KSCE Journal of Civil and Environmental Engineering Research
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    • v.31 no.1D
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    • pp.127-132
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    • 2011
  • Load managing method on road became necessary since overloaded vehicles occur damage on road facilities and existing systems for preventing this damage still show many problems. Accordingly, efficient managing system for preventing overloaded vehicles could be organized by using the road itself as a scale by applying genetic algorithm to analyze the load and the drive information of vehicles. First of all, accurate analysis of load using the behavior of road itself is needed for solving illegal axle manipulation problems of overloaded vehicles and for installing intelligent embedded load analyzing system. Accordingly in this study, to use the behavior of road, the transformation was measured by installing underground box type indoor model and indoor experiment was held using genetic algorithm and 10% error were checked.

Construction and Calibration Test of a Transmission-type Circular Polariscope for Photoelastic Stress Measurement (광탄성 응력측정을 위한 투과형 원형편광기 제작 및 시험)

  • Baek, T.H.;Koh, S.K.;Na, E.G.;Kim, J.S.;Cha, B.S.;Cho, C.H.
    • Proceedings of the KSME Conference
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    • 2001.11a
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    • pp.437-441
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    • 2001
  • This paper describes the construction of a circular polariscope. Generally, a circular polariscope contains four optical elements and a light source. The first element following the light source is called the polarizer. It converts the ordinary light into plane-polarized light. The second element is a quarter wave plate which converts the plane-polarized light into circularly polarized light. Following the quarter wave plate, a specimen made of transparent photoelastic material is located in a loading device. The second quarter wave plate is set and the last element is the analyzer. These polarizing elements, two quarter wave plates and two polarizing filters, were purchased from the USA. Frames and other structures for holding polarizing filters were machined and assembled to be rotated. Light box, which include four incandescent lamps and two sodium-vapor lamps, was made. In order to proof the function of the newly built polariscope, Tardy compensation test was applied to a rectangular shaped specimen made of poly-carbonate material (PSM1). The error of the fringe constant, which was measured by the newly built polariscope, was within 4.4 percent compared to the standard value of this material. It is possible to make a good quality of polariscope if accurate polarizing filters will be used.

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Global Sensitivity Analysis of Joints for Plug-in Digital Framework (플러그인 디지털 프레임웍을 위한 연결부 전역민감도 해석)

  • Lee, Dooho;Won, Young-Woo;Kwon, Jong-Hyun
    • Transactions of the Korean Society of Mechanical Engineers A
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    • v.40 no.5
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    • pp.483-488
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    • 2016
  • Plug-In Digital Framework is a system response analysis tool that is employed when system components are composed of black-box modules. Generally, the dynamic characteristics of joints between the system components significantly affect system responses, and they lead to displacement- and frequency-dependent stiffness and loss factor. Thus, the sensitivity of each joint parameters should be estimated from a global perspective. In this study, we introduce a global sensitivity analysis procedure under the Plug-In Digital Framework. To efficiently calculate the system responses, we introduce the frequency response function (FRF)-based substructuring method. Using the random balance designs (RBD), we generate the system responses and estimate the global first-order sensitivities for each joint stiffness. We apply the proposed global sensitivity analysis method to an interior noise problem of a passenger car, and we evaluate the efficiency of the global sensitivity analysis method.

Effects of Rearing Density on Growth of the Polychaete Rockworm Marphysa sanguinea

  • Parandavar, Hossein;Kim, Kyeong-Hun;Kim, Chang-Hoon
    • Fisheries and Aquatic Sciences
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    • v.18 no.1
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    • pp.57-63
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    • 2015
  • Effects of rearing density on growth and survival of the polychaete rockworm Marphysa sanguinea have been investigated in order to develop rearing techniques for this species. This study was examined over a nine-month period in the Fisheries Science and Technology Center of Pukyong National University. Three rockworm densities, 500, 1,000 and $2,000worms{\cdot}m^{-2}$ with weight ranges of < 0.5 g, 0.6-1.5 g, and 1.6-2.5 g, and the no feed control treatment, were stocked in triplicate $0.10m^2$ boxes with sand bottoms. Growth rates were checked with 15 randomly sampled rockworms from each box at months 3, 6 and 9. Results showed that SGRs in all treatments were higher during the first period (0-3 months) than the second (3-6 months) and third periods (6-9 months) for all treatment densities, while SGRs decreased with increasing density. However, survival and growth of worms at high density was not better than low density, but daily biomass production in medium and high density groups was $6.28gm^{-2}day^{-1}$ for the rockworms of 0.6-1.5 g with $2,000inds{\cdot}m^{-2}$, and $12.6gm^{-2}day^{-1}$ for group between 1.6-2.5 g with $2,000inds{\cdot}m^{-2}$, and $14.7gm^{-2}day^{-1}$ for the group of individuals <0.5 g with $1,000inds{\cdot}m^{-2}$. Results showed that M. sanguinea can be one of the most suitable species to commercially exploit in a farming system. In particular, specified densities permit elevated pure production.

Style Analysis on : Barry Salt's Statistical Analysis (<지슬>의 스타일 분석: 배리 솔트의 통계적 분석을 중심으로)

  • Kim, Jung-Ho;Kim, Hak-Min
    • The Journal of the Korea Contents Association
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    • v.14 no.10
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    • pp.486-505
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    • 2014
  • is the winner of Grand Jury Prize in World Cinema-Dramatic at Sundance Film Festival 2013 and made high Box Office performance in Korean Indie Film distribution. tells the story of the horrific massacre of 30,000 civilians at Jeju Island in Korea in 1948. Civilians were killed by Korean Army under the supervision of U. S. Army and by the Korean Communist partisan. From the Statistical Style Analysis, has a lot in common with Ming-liang Tsai's movies and similar traits of art cinema. According to David Boardwell, Art Cinema is intensify the traditional continuity through selection and concentration rather than become the Alternative, Oppositional film practice of Mainstream Classical Hollywood Cinema.

Variations of Air Temperature, Relative Humidity and Pressure in a Low Pressure Chamber for Plant Growth (식물생장용 저압챔버 내의 기온, 상대습도 및 압력의 변화)

  • Park, Jong-Hyun;Kim, Yong-Hyeon
    • Journal of Bio-Environment Control
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    • v.18 no.3
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    • pp.200-207
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    • 2009
  • This study was conducted to analyze the variations of air temperature, relative humidity and pressure in a low pressure chamber for plant growth. The low pressure chamber was composed of an acrylic cylinder, a stainless plate, a mass flow controller, an elastomer pressure controller, a read-out-box, a vacuum pump, and sensors of air temperature, relative humidity, and pressure. The pressure leakage in the low pressure chamber was greatly affected by the material and connection method of tubes. The leakage rate in the low pressure chamber with the welding of the stainless tubes and a plate decreased by $0.21kPa{\cdot}h^{-1}$, whereas the leakage in the low pressure chamber with teflon tube and rubber O-ring was given by $1.03kPa{\cdot}h^{-1}$. Pressure in the low pressure chamber was sensitively fluctuated by the air temperature inside the chamber. An elastomer pressure controller was installed to keep the pressure in the low pressure chamber at a setting value. However, inside relative humidity at dark period increased to saturation level.. Two levels (25 and 50kPa) of pressure and two levels (500 and 1,000sccm) of mass flow rate were provided to investigate the effect of low pressure and mass flow rate on relative humidity inside the chamber. It was concluded that low setting value of pressure and high mass flow rate of mixed gas were the effective methods to control the pressure and to suppress the excessive rise of relative humidity inside the chamber.

Forskolin-Induced Stimulation of RGS2 mRNA in C6 Astrocytoma Cells

  • Kim Sung-Dae;Cho Jae-Youl;Park Hwa-Jin;Kim Sang-Keun;Rhee Man-Hee
    • Biomedical Science Letters
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    • v.12 no.3
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    • pp.131-137
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    • 2006
  • RGS is a negative regulator of G-protein signaling and can be identified by the presence of a conserved $120{sim}125$ amino acid motif, which is referred to as the RGS box. A number of RGSs are induced in response to a wide variety of stimuli. Increased levels of RGSs lead to significant decreases in GPCR responsiveness. To obtain further evidence of a role of RGS proteins in rat C6 astrocytoma cells, we first determined the expression profile of RGS-specific mRNA in C6 cells using reverse transcription-polymerase chain reaction (RT-PCR) with a poly dT18 primer and transcript-specific primers. We found that RGS2, RGS3, RGS6, RGS9, RGS10, RGS12, and RGS16 were differentially expressed in C6 astrocytoma cells. The highest expression rate was found for RGS3, followed by RGS16, RGS10 and RGS9, whereas the expression level for RGS2 was barely detectable. We next assessed whether forskolin regulated the expression of RGSs expressed in C6 astrocytoma cells. The present study found that forskolin dose-dependently stimulated the expression of RGS2 transcripts. This up-regulation of RGS2 gene was abrogated by H-89, potent and broad-spectrum protein kinase A (PKA) inhibitors. Actinomycin D completely inhibited the up-regulation of RGS2 gene induced by forskolin $(10{\mu}M)$, indicating that the regulation of RGS2 gene is controlled at the transcriptional level. In addition, forskolin did significantly activate transcriptional cAMP response element (CRE) in either HEK 293 cells or C6 cells and did not modulate the $NF-{\kappa}B$ and AP-l activity as measured by luciferase reporter gene assay. Finally, forskolin induced the expression of RGS2 mRNA in C6 astrocytoma cells, which depend on the PKA pathway and CRE transcriptional pathways.

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Anti-septic effects of dabrafenib on HMGB1-mediated inflammatory responses

  • Jung, Byeongjin;Kang, Hyejin;Lee, Wonhwa;Noh, Hyun Jin;Kim, You-Sun;Han, Min-Su;Baek, Moon-Chang;Kim, Jaehong;Bae, Jong-Sup
    • BMB Reports
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    • v.49 no.4
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    • pp.214-219
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    • 2016
  • A nucleosomal protein, high mobility group box 1 (HMGB1) is known to be a late mediator of sepsis. Dabrafenib is a B-Raf inhibitor and initially used for the treatment of metastatic melanoma therapy. Inhibition of HMGB1 and renewal of vascular integrity is appearing as an engaging therapeutic strategy in the administration of severe sepsis or septic shock. Here, we examined the effects of dabrafenib (DAB) on the modulation of HMGB1-mediated septic responses. DAB inhibited the release of HMGB1 and downregulated HMGB1-dependent inflammatory responses by enhancing the expressions of cell adhesion molecules (CAMs) in human endothelial cells. In addition, treatment with DAB inhibited the HMGB1 secretion by CLP and sepsis-related mortality and pulmonary injury. This study demonstrated that DAB could be alternative therapeutic options for sepsis or septic shock via the inhibition of the HMGB1 signaling pathway.