• Title/Summary/Keyword: Kappa analysis

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Inhibition of Inducible Nitric Oxide Synthase Expression by YS 49, a Synthetic Isoquinoline Alkaloid, in ROS 17/2.8 Cells Activated with $TNF-{\alpha},\;IFN-{\gamma}$ and LPS

  • Kang, Young-Jin;Kang, Sun-Young;Lee, Young-Soo;Park, Min-Kyu;Kim, Hye-Jung;Seo, Han-Geuk;Lee, Jae-Heun;YunChoi, Hye-Sook;Chang, Ki-Churl
    • The Korean Journal of Physiology and Pharmacology
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    • v.8 no.5
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    • pp.273-280
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    • 2004
  • Nitric oxide (NO) has been suggested to act as a mediator of cytokine-induced effects of turn over of bone. Activation of the inducible nitric oxide synthase (iNOS) by inflammation has been related with apoptotic cell death in osteoblast. YS 49, a synthetic isoquinoline alkaloid, inhibits NO production in macrophages activated with cytokines. In the present study, we investigated the molecular mechanism of YS 49 to inhibit iNOS expression in ROS 17/2.8 cells, which were activated with combined treatment of inflammatory cytokines $(TNF-{\alpha},\;IFN-{\gamma})$ and lipopolysaccharide (LPS). Results indicated that YS 49 concentration-dependently reduced iNOS mRNA and protein expression, as evidenced by Northern and Western blot analysis, respectively. The underlying mechanism by which YS 49 suppressed iNOS expression was not to affect iNOS mRNA stability but to inhibit activation and translocation of $NF-_kB$ by preventing the degradation of its inhibitory protein $I_kB_{\alpha}$. As expected, YS 49 prevented NO-induced apoptotic cell death by sodium nitroprusside. Taken together, it is concluded that YS 49 inhibits iNOS expression by interfering with degradation of phosphorylated inhibitory $_kB_{\alpha}\;(p-I_kB_{\alpha})$. These actions may be beneficial for the treatment of inflammation of the joint, such as rheumatoid arthritis.

Accuracy and reliability of 2-dimensional photography versus 3-dimensional soft tissue imaging

  • Ayaz, Irem;Shaheen, Eman;Aly, Medhat;Shujaat, Sohaib;Gallo, Giulia;Coucke, Wim;Politis, Constantinus;Jacobs, Reinhilde
    • Imaging Science in Dentistry
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    • v.50 no.1
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    • pp.15-22
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    • 2020
  • Purpose: This study was conducted to objectively and subjectively compare the accuracy and reliability of 2-dimensional(2D) photography and 3-dimensional(3D) soft tissue imaging. Materials and Methods: Facial images of 50 volunteers(25 males, 25 females) were captured with a Nikon D800 2D camera (Nikon Corporation, Tokyo, Japan), 3D stereophotogrammetry (SPG), and laser scanning (LS). All subjects were imaged in a relaxed, closed-mouth position with a normal smile. The 2D images were then exported to Mirror® Software (Canfield Scientific, Inc, NJ, USA) and the 3D images into Proplan CMF® software (version 2.1, Materialise HQ, Leuven, Belgium) for further evaluation. For an objective evaluation, 2 observers identified soft tissue landmarks and performed linear measurements on subjects' faces (direct measurements) and both linear and angular measurements on all images(indirect measurements). For a qualitative analysis, 10 dental observers and an expert in facial imaging (subjective gold standard) completed a questionnaire regarding facial characteristics. The reliability of the quantitative data was evaluated using intraclass correlation coefficients, whereas the Fleiss kappa was calculated for qualitative data. Results: Linear and angular measurements carried out on 2D and 3D images showed excellent inter-observer and intra-observer reliability. The 2D photographs displayed the highest combined total error for linear measurements. SPG performed better than LS, with borderline significance (P=0.052). The qualitative assessment showed no significant differences among the 2D and 3D imaging modalities. Conclusion: SPG was found to a reliable and accurate tool for the morphological evaluation of soft tissue in comparison to 2D imaging and laser scanning.

Effects of all-trans retinoic acid on expression of Toll-like receptor 5 on immune cells (All-trans retinoic acid가 면역세포의 Toll-like receptor 5 발현에 미치는 영향)

  • Kim, Ki-Hyung;Park, Sang-Jun
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.36 no.6
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    • pp.481-489
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    • 2010
  • Introduction: TLR-5, a member of the toll-like receptor (TLR) family, is a element of the type I transmembrane receptors, which are characterized by an intracellular signaling domain homolog to the interleukin-1 receptor. These receptors recognize microbial components, particularly bacterial flagellin. All-trans retinoic acid (atRA, tretinoin), a natural metabolite of vitamin A, acts as a growth and differentiation factor in many tissues, and is also needed for immune functions. In this study, THP-1 human macrophage-monocytes were used to examine the mechanisms by which atRA regulated the expression of TLR-5. Because the molecular mechanism underlying this regulation at the transcriptional level is also unclear, this study examined which putative transcription factors are responsible for TLR-5 expression by atRA in immune cells. Materials and Methods: This study examined whether atRA induces the expression of TLR-5 in THP-1 cells using reverse transcription-polymerase chain reaction (RT-PCR), and which transcription factors are involved in regulating the TLR-5 promoter in RAW264.7 cells using a reporter assay system. Western blot analysis was used to determine which signal pathway is involved in the expression of TLR-5 in atRA-treated THP-1 cells. Results: atRA at a concentration of 10 nM greatly induced the expression of TLR-5 in THP-1 cells. Human TLR-5 promoter contains three Sp-1/GC binding sites around -50 bp and two NF-kB binding sites at -380 bp and -160 bp from the transcriptional start site of the TLR-5 gene. Sp-1/GC is primarily responsible for the constitutive TLR-5 expression, and may also contribute to NF-kB at -160 bp to induce TLR-5 after atRA stimulation in THP-1 cells. The role of NF-kB in TLR-5 expression was further confirmed by inhibitor pyrrolidine dithiocarbamate (PDTC) experiments, which greatly reduced the TLR-5 transcription by 70-80%. Conclusion: atRA induces the expression of the human TLR-5 gene and NF-kB is a critical transcription factor for the atRA-induced expression of TLR-5. Accordingly, it is conceivable that retinoids are required for adequate innate and adaptive immune responses to agents of infectious diseases. atRA and various synthetic retinoids have been used therapeutically in human diseases, such as leukemia and other cancers due to the antiproliferative and apoptosis inducing effects of retinoids. Therefore, understanding the molecular regulatory mechanism of TLR-5 may assist in the design of alternative strategies for the treatment of infectious diseases, leukemia and cancers.

Computer Simulation for the Thermal Analysis of the Energy Storage Board (에너지 축열보드 열해석을 위한 컴퓨터 수치해석)

  • 강용혁;엄태인;곽희열
    • Journal of Energy Engineering
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    • v.8 no.2
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    • pp.224-232
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    • 1999
  • Latent heat storage system using micro-encapsuled phase change material is effective method for floor heating of house and building. The temperature profile in capsule block and flow rate of hot water are important parameters for the development of heat storage system. In the present study, a mathematical model based on 3-D, non-steady state, Navier-Stokes equations, scalar conservation equations and turbulence model ($\kappa$-$\varepsilon$), is used to predict the temperature profiles in capsule and the velocity vectors in hot water pipe. The multi-block grids and fine grids embedding are used to join the circle in hot water pipe and square in capsule block. The phase change process of the capsule is quite complex not only because the size of phase change material is very small, but also because phase change material is mixed with the cement to form thermal storage block. In calculation, it's assumed that the phenomena of phase change is limited only the thermal properties of phase change material and the change of boundary is not happened in capsule. The purpose of this study is to calculate the temperature profiles in capsule block and velocity vectors in hot water pipe using the numerical calculation. Two kinds of thermal boundary condition were considered, the first (case 1) is the adiabatic condition for the both outside surfaces of the wall, the second (case 2) is the case in which one surface is natural convection with atmosphere and another surface is adaibatic. Calculation results are shown that the temperature profile in capsule block for case 1 is higher than that for case 2 due to less heat loss in adaibatic surface. Specially, in the domain of near Y=0, the difference of temperature is greater in case 1 than in case 2. The detailed experimental data of capsule block on the temperature profile and the thermal properties such as specific heat and coefficient of heat transfer with the various temperature are required to predict more exact phenomena of heat transfer.

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Effects of High Pressure and Binding Agents on the Quality Characteristics of Restructured Pork (초고압처리 및 결착제 첨가가 재구성 돈육의 품질특성에 미치는 효과)

  • Choi, Ye-Chul;Jung, Kyung-Hun;Chun, Ji-Yeon;Choi, Mi-Jung;Hong, Geun-Pyo
    • Food Science of Animal Resources
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    • v.33 no.5
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    • pp.664-671
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    • 2013
  • The objective of this study was to investigate the effect of high pressure treatment and type of binding agents on the quality characteristics of restructured pork. For binding agents, 2% (w/w) isolated soy protein (SP), 0.5% (w/w) wheat flour (WF) and 0.5% (w/w) ${\kappa}$-carrageenan (KC) were incorporated into meat batter with or without 0.5% (w/w) glucono-${\delta}$-lactone (GdL). The restructured pork was pressurized at varying pressure levels (0.1-450 MPa) for 3 min under ambient temperature and thermal treated at $75^{\circ}C$ for 30 min. As quality parameters of restructured pork, pH, water binding properties, instrumental color and texture profile analysis were determined and compared with control (C, no binder). For type of binders, SP exhibited the best water binding properties, however, the impact on textural properties were lesser than KC and WF. The addition of GdL decreased the pH of restructured pork down to 0.4 unit, while high pressure processing prevented the moisture loss caused from pH decrease by GdL. In particular, meat restructuring efficiency of SP as a binder improved under the presence of GdL. Therefore, the present study demonstrated the potential advantages of low amount of GdL (0.5%, w/w) combined with protein based binder (SP) and high pressure processing in restructuring meat particles.

High fat diet-induced obesity leads to proinflammatory response associated with higher expression of NOD2 protein

  • Kim, Min-Soo;Choi, Myung-Sook;Han, Sung-Nim
    • Nutrition Research and Practice
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    • v.5 no.3
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    • pp.219-223
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    • 2011
  • Obesity has been reported to be associated with low grade inflammatory status. In this study, we investigated the inflammatory response as well as associated signaling molecules in immune cells from diet-induced obese mice. Four-week-old C57BL mice were fed diets containing 5% fat (control) or 20% fat and 1% cholesterol (HFD) for 24 weeks. Splenocytes ($1{\times}10^7$ cells) were stimulated with $10\;{\mu}g/mL$ of lipopolysaccharide (LPS) for 6 or 24 hrs. Production of interleukin (IL)-$1{\beta}$, IL-6, and TNF-${\alpha}$ as well as protein expression levels of nucleotide-binding oligomerization domain (NOD)2, signal transducer and activator of transcription (STAT)3, and pSTAT3 were determined. Mice fed HFD gained significantly more body weight compared to mice fed control diet ($28.2{\pm}0.6$ g in HFD and $15.4{\pm}0.8$ g in control). After stimulation with LPS for 6 hrs, production of IL-$1{\beta}$ was significantly higher (P=0.001) and production of tumor necrosis factor (TNF)-${\alpha}$ tended to be higher (P < 0.064) in the HFD group. After 24 hrs of LPS stimulation, splenocytes from the HFD group produced significantly higher levels of IL-6 ($10.02{\pm}0.66$ ng/mL in HFD and $7.33{\pm}0.56$ ng/mL in control, P=0.005) and IL-$1{\beta}$ ($121.34{\pm}12.72$ pg/mL in HFD and $49.74{\pm}6.58$ pg/mL in control, P < 0.001). There were no significant differences in the expression levels of STAT3 and pSTAT3 between the HFD and the control groups. However, the expression level of NOD2 protein as determined by Western blot analysis was 60% higher in the HFD group compared with the control group. NOD2 contributes to the induction of inflammation by activation of nuclear factor ${\kappa}B$. These findings suggest that diet-induced obesity is associated with increased inflammatory response of immune cells, and higher expression of NOD2 may contribute to these changes.

Melanin extract from Gallus gallus domesticus promotes proliferation and differentiation of osteoblastic MG-63 cells via bone morphogenetic protein-2 signaling

  • Yoo, Han-Seok;Chung, Kang-Hyun;Lee, Kwon-Jai;Kim, Dong-Hee;An, Jeung Hee
    • Nutrition Research and Practice
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    • v.11 no.3
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    • pp.190-197
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    • 2017
  • BACKGROUND/OBJECTIVES: Gallus gallus domesticus (GD) is a natural mutant breed of chicken in Korea with an atypical characterization of melanin in its tissue. This study investigated the effects of melanin extracts of GD on osteoblast differentiation and inhibition of osteoclast formation. MATERIALS/METHODS: The effects of the melanin extract of GD on human osteoblast MG-63 cell differentiation were examined by evaluating cell viability, osteoblast differentiation, and expression of osteoblast-specific transcription factors such as bone morphogenetic protein 2 (BMP-2), small mothers against decapentaplegic homologs 5 (SMAD5), runt-related transcription factor 2 (RUNX2), osteocalcin and type 1 collagen (COL-1) by reverse transcription-polymerase chain reaction and western blotting analysis. We investigated the inhibitory effect of melanin on the osteoclasts formation through tartrate-resistant acid phosphatase (TRAP) activity and TRAP stains in Raw 264.7 cell. RESULTS: The melanin extract of GD was not cytotoxic to MG-63 cells at concentrations of $50-250{\mu}g/mL$. Alkaline phosphatase (ALP) activity and bone mineralization of melanin extract-treated cells increased in a dose-dependent manner from 50 to $250{\mu}g/mL$ and were 149% and 129% at $250{\mu}g/mL$ concentration, respectively (P < 0.05). The levels of BMP-2, osteocalcin, and COL-1 gene expression were significantly upregulated by 1.72-, 4.44-, and 2.12-fold in melanin-treated cells than in the control cells (P < 0.05). The levels of RUNX2 and SMAD5 proteins were higher in melanin-treated cells than in control vehicle-treated cells. The melanin extract attenuated the formation of receptor activator of nuclear factor kappa-B ligand-induced TRAP-positive multinucleated RAW 264.7 cells by 22%, and was 77% cytotoxic to RAW 264.7 macrophages at a concentration of $500{\mu}g/mL$. CONCLUSIONS: This study provides evidence that the melanin extract promoted osteoblast differentiation by activating BMP/SMADs/RUNX2 signaling and regulating transcription of osteogenic genes such as ALP, type I collagen, and osteocalcin. These results suggest that the effective osteoblastic differentiation induced by melanin extract from GD makes it potentially useful in maintaining bone health.

Analysis of Land Use Change Using RCP-Based Dyna-CLUE Model in the Hwangguji River Watershed (RCP 시나리오 기반 Dyna-CLUE 모형을 이용한 황구지천 유역의 토지이용변화 분석)

  • Kim, Jihye;Park, Jihoon;Song, Inhong;Song, Jung-Hun;Jun, Sang Min;Kang, Moon Seong
    • Journal of Korean Society of Rural Planning
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    • v.21 no.2
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    • pp.33-49
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    • 2015
  • The objective of this study was to predict land use change based on the land use change scenarios for the Hwangguji river watershed, South Korea. The land use change scenario was derived from the representative concentration pathways (RCP) 4.5 and 8.5 scenarios. The CLUE (conversion of land use and its effects) model was used to simulate the land use change. The CLUE is the modeling framework to simulate land use change considering empirically quantified relations between land use types and socioeconomic and biophysical driving factors through dynamical modeling. The Hwangguji river watershed, South Korea was selected as study area. Future land use changes in 2040, 2070, and 2100 were analyzed relative to baseline (2010) under the RCP4.5 and 8.5 scenarios. Binary logistic regressions were carried out to identify the relation between land uses and its driving factors. CN (Curve number) and impervious area based on the RCP4.5 and 8.5 scenarios were calculated and analyzed using the results of future land use changes. The land use change simulation of the RCP4.5 scenario resulted that the area of urban was forecast to increase by 12% and the area of forest was estimated to decrease by 16% between 2010 and 2100. The land use change simulation of the RCP8.5 scenario resulted that the area of urban was forecast to increase by 16% and the area of forest was estimated to decrease by 18% between 2010 and 2100. The values of Kappa and multiple resolution procedure were calculated as 0.61 and 74.03%. CN (III) and impervious area were increased by 0-1 and 0-8% from 2010 to 2100, respectively. The study findings may provide a useful tool for estimating the future land use change, which is an important factor for the future extreme flood.

Gene Structure and Altered mRNA Expression of Metallothionein in Response to Metal Exposure and Thermal Stress in Miho Spine Loach Cobitis choii (Cobitidae; Cypriniformes) (미호종개 metallothionein 유전자의 구조 및 중금속 노출과 고온 자극에 대한 MT mRNA의 발현 특징 분석)

  • Lee, Sang-Yoon;Nam, Yoon-Kwon
    • Korean Journal of Ichthyology
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    • v.23 no.1
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    • pp.61-69
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    • 2011
  • Gene and promoter structures of metallothionein(MT) from Miho spine loach (Cobitis choii; Cypriniformes) were characterized, and the transcriptional responses to experimental exposures to heavy metals and heat stress were examined. The C. choii metallothionein displayed well-conserved features of teleostean metallothioneins at gDNA, mRNA and amino acid levels. Bioinformatic analysis predicted that the C. choii MT regulatory region potentially possessed various motifs or elements targeted by various transcription factors associated with metal-coordinating regulation (e.g., metal transcription factor-1), immune responses (e.g., nuclear factor kappa B), and thermal modulations (e.g., heat shock factor). Acute heavy-metal exposures to 0.5 or $1.0\;{\mu}M$ of cadmium (Cd), copper (Cu), manganese (Mn), nickel (Ni) or zinc (Zn) showed that MT transcription was significantly stimulated by Cd (9.6-fold relative to non-exposed control) and Cu (10.4-fold), only moderately by Mn (2.4-fold), but hardly by Ni and Zn. Elevation of water temperature from $25^{\circ}C$ to $31^{\circ}C$ caused a rapid modulation of MT mRNAs toward upregulation to 9.5-fold; however, afterward the elevated mRNA level slightly decreased during further incubation at $31^{\circ}C$ for 6 h. Results from this study suggest that MT-based expression assay could be a useful basis for better understanding the metal- and/or heat-caused stresses in this endangered fish species.

Study of threshold and opacity in three-dimensional CT volume rendering of oral and maxillofacial area (구강악안면영역의 3차원 CT 영상 재형성시 역치 및 불투명도에 대한 연구)

  • Choi, Mun-Kyung;Lee, Sam-Sun;Huh, Kyung-Hoe;Yi, Won-Jin;Choi, Soon-Chul
    • Imaging Science in Dentistry
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    • v.39 no.1
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    • pp.13-18
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    • 2009
  • Purpose: This study was designed to determine a proper threshold value and opacity in three-dimensional CT volume rendering of oral and maxillofacial area. Materials and Methods: Three-dimensional CT data obtained from 50 persons who were done orthognatic surgery in department of oral and maxillofacial radiology of Seoul National University retrospectively. 12 volume rendering post-processing protocols of combination of threshold(100HU, 150HU, 221HU, 270HU) and opacity (58%, 80%, 90%) were applied. Five observers independently evaluated image quality using a five-point range scale. The results were analyzed by receiver operating characteristic curves, ANOVA and Kappa value. And three oromaxillofacial surgeons chose the all images that they thought proper clinically in the all of images. Results: Analysis using ROC curves revealed the area under each curve which indicated a diagnostic accuracy. The highest diagnostic accuracy appear with 100HU and 58% opacity. and the lowest diagnostic accuracy appear with 221HU and 58% opacity that are being used protocol in department of oral and maxillofacial radiology of Seoul National University. But, no statistically significant difference was noted between any of the protocols. And the number of proper images clinically that chosen by three oromaxillofacial surgeons is the largest in the cases of protocol 8 (221HU, opacity 80%) and protocol 11 (270HU, opacity 80%) in one after the other. Conclusion: Threshold and opacity in volume rendering can be controled easily and these can be causes of making an diagnostic accuracy. So we need to select proper values of these factors.

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