• Title/Summary/Keyword: 3-D Mechanism

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The Mechanisms of Resistance to TNF in TNF-Sensitive Cancer Cells Transfected with TNF-$\alpha$ Gene Using Retroviral Vector (암세포에서 Retroviral Vector를 이용한 종양괴사인자 유전자 이입후 획득된 종양괴사인자 내성의 기전)

  • Lee, Hyuk-Pyo;Oh, Yeon-Mok;Yoo, Chul-Gyu;Kim, Young-Whan;Shim, Young-Soo;Han, Sung-Koo
    • Tuberculosis and Respiratory Diseases
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    • v.44 no.3
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    • pp.547-558
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    • 1997
  • Background : Tumor necrosis factor(TNF) has been considered as an important candidate for cancer gene therapy based on its potent anti-tumor activity. However, since the efficiency of current techniques of gene transfer is not satisfactory, the majorities of current protocols is aiming the in vitro gene transfer to cancer cells and re-introducing genetically modified cancer cells to hoot. In previous study, it was shown that TNF-sensitive cancer cells transfected with TNF-$\alpha$ cDNA would become highly resistant to TNF. Understanding the mechanisms of TNF-resistance in TNF-$\alpha$ gene transfected cancer cells would be an important step for improving the efficacy of cancer gene therapy as well as for better understandings of tumor biology. This study was designed to evaluate the role of new protective protein synthesis in the acquired resistance to TNF of TNF-$\alpha$ gene transfected cancer cells. Method : We transfected TNF-$\alpha$ c-DNA to WEHI164, a murine fibrosarcoma cell line, using retroviral vector(pLT12SN(TNF)) and confirm the expression of TNF with PCR, ELISA, MIT assay. Then we determined the TNF resistance of TNF gene transfected cells(WEHI164-TNF) and the changes of TNF sensitivities after treatments with actinomycin D(transcription inhibitor) and cycloheximide ( translation inhibitor). Results : WEHI164 which was sensitive to TNF became resistant to TNF after being transfected with TNF-$\alpha$ gene and the resistance to TNF was partially reversed after treatment with actinomycin D, but not with cycloheximide. Conclusion : The acquired resistance to TNF after TNF-$\alpha$ gene transfection may be associated with synthesis of some protective proteins.

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G Protein-Coupled Receptor Signaling in Gastrointestinal Smooth Muscle

  • Sohn, Uy-Dong;Kim, Dong-Seok;Murthy, Karnam S.
    • The Korean Journal of Physiology and Pharmacology
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    • v.5 no.4
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    • pp.287-297
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    • 2001
  • Contraction of smooth muscle is initiated by an increase in cytosolic $Ca^{2+}$ leading to activation of $Ca^{2+}$/ calmodulin-dependnet myosin light chain (MLC) kinase and phosphorylation of MLC. The types of contraction and signaling mechanisms mediating contraction differ depending on the region. The involvement of these different mechanisms varies depending on the source of $Ca^{2+}$ and the kinetic of $Ca^{2+}$ mobilization. $Ca^{2+}$ mobilizing agonists stimulate different phospholipases $(PLC-{\beta},\;PLD\;and\;PLA_2)$ to generate one or more $Ca^{2+}$ mobilizing messengers $(IP_3\;and\;AA),$ and diacylglycerol (DAG), an activator of protein kinase C (PKC). The relative contributions of $PLC-{\beta},\;PLA_2$ and PLD to generate second messengers vary greatly between cells and types of contraction. In smooth muscle cell derived form the circular muscle layer of the intestine, preferential hydrolysis of $PIP_2$ and generation of $IP_3$ and $IP_3-dependent\;Ca^{2+}$ release initiate the contraction. In smooth muscle cells derived from longitudinal muscle layer of the intestine, preferential hydrolysis of PC by PLA2, generation of AA and AA-mediated $Ca^{2+}$ influx, cADP ribose formation and $Ca^{2+}-induced\;Ca^{2+}$ release initiate the contraction. Sustained contraction, however, in both cell types is mediated by $Ca^{2+}-independent$ mechanism involving activation of $PKC-{\varepsilon}$ by DAG derived form PLD. A functional linkage between $G_{13},$ RhoA, ROCK, $PKC-{\varepsilon},$ CPI-17 and MLC phosphorylation in sustained contraction has been implicated. Contraction of normal esophageal circular muscle (ESO) in response to acetylcholine (ACh) is linked to $M_2$ muscarinic receptors activating at least three intracellular phospholipases, i.e. phosphatidylcholine-specific phospholipase C (PC-PLC), phospholipase D (PLD) and the high molecular weight (85 kDa) cytosolic phospholipase $A_2\;(cPLA_2)$ to induce phosphatidylcholine (PC) metabolism, production of diacylglycerol (DAG) and arachidonic acid (AA), resulting in activation of a protein kinase C (PKC)-dependent pathway. In contrast, lower esophageal sphincter (LES) contraction induced by maximally effective doses of ACh is mediated by muscarinic $M_3$ receptors, linked to pertussis toxin-insensitive GTP-binding proteins of the $G_{q/11}$ type. They activate phospholipase C, which hydrolyzes phosphatidylinositol bisphosphate $(PIP_2),$ producing inositol 1, 4, 5-trisphosphate $(IP_3)$ and DAG. $IP_3$ causes release of intracellular $Ca^{2+}$ and formation of a $Ca^{2+}$-calmodulin complex, resulting in activation of myosin light chain kinase and contraction through a calmodulin-dependent pathway.

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Photocatalytic Degradation and Detoxification of Bisphenol A Using TiO2 Nanoparticles (TiO2 나노입자 광촉매 반응에 의한 비스페놀 A의 분해 제거 및 독성 저감)

  • Jo, A-Yeong;Jung, Jinho
    • Ecology and Resilient Infrastructure
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    • v.2 no.4
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    • pp.330-336
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    • 2015
  • Photocatalytic degradation of bisphenol A (BPA) in aqueous solution was investigated using $TiO_2$ nanoparticles (Degussa P25) in this study. After a 3 hr photocatalytic reaction (${\lambda}=365nm$ and $I=3mW\;cm^{-2}$, $[TiO_2]=2.0g\;L^{-1}$), 98% of BPA ($1.0{\times}10^{-5}M$) was degraded and 89% of the total organic carbon was removed. In addition, BPA degradation by photolytic, hydrolytic and adsorption reactions was found to be 2%, 5% and 13%, respectively. The reaction rate of BPA degradation by photocatalysis decreased with increasing concentration of methanol that is used as a hydroxyl radical scavenger. This indicates that the reaction between BPA and hydroxyl radical was the key mechanism of BPA degradation. The pseudo-first-order reaction rate constant for this reaction was determined to be $7.94{\times}10^{-4}min^{-1}$, and the time for 90% BPA removal was found to be 25 min. In addition, acute toxicity testing using Daphnia magna neonates (< 24 h old) was carried out to evaluate the reduction of BPA toxicity. Acute toxicity (48 hr) to D. magna was decreased from 2.93 TU (toxic unit) to non-toxic after photocatalytic degradation of BPA for 3 hr. This suggests that there was no formation of toxic degradation products from BPA photocatalysis.

Deposition and Characteristics of TiN Thin Films by Atomic Layer Epitaxy (ALE 법에 의한 TiN 박막의 증착 및 특성)

  • Kim, Dong-Jin;Jung, Young-Bae;Lee, Myung-Bok;Lee, Jung-Hee;Lee, Yong-Hyun;Hahm, Sung-Ho;Lee, Jong-Hwa
    • Journal of the Institute of Electronics Engineers of Korea SD
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    • v.37 no.6
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    • pp.43-49
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    • 2000
  • The TiN thin films were deposited by ALE(atomic layer epitaxy) on (100) silicon substrate. The TiN thin films were characterized by means of XRD, 4-point probe, AFM, AES and SEM. TEMAT(terakis(ethyl methy lamino)titanium) and $NH_3$ were injected into the reactor in sequence of TEMAT-$N_2-NH_3-N_2$ to ensure a saturated surface reaction. As a result, the depostion rate of the TiN film was controlled by self-limiting growth mechanism at temperature range form 150 to 220 $^{\circ}C$. Deposited TiN films, all of which show amorphous structure, had a fixed deposition rate of 4.5 ${\AA}$/cycle. The resistivity of 210 ~ 230 ${\mu}{\Omega}{\cdot}$cm and the surface r.m.s. roughness of 7.9 ~ 9.3 ${\AA}$ were measured. When TiN film of 2000 ${\AA}$ were deposited, a excellent step coverage were observed in a trench structure of 0.43${\mu}m$ contacts with 6:1 aspect ratio.

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A Study on the Three Dimensional Finite Element Analysis for the Tunnel Reinforced by Umbrella Arch Method (Umbrella Arch 공법이 적용된 터널의 3차원 유한요소 해석에 관한 연구)

  • 김창용;배규진;문현구;최용기
    • Tunnel and Underground Space
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    • v.8 no.3
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    • pp.209-225
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    • 1998
  • Recently, Umbrella Arch Method(UAM), one of the auxiliary techniques for tunnelling, is used to reinforce the ground and improve stability of tunnel face. Because UAM combines the advantages of a modern forepoling system with the grouting injection method, this technique has been applied in subway, road and utility tunnel sites for the last few years in Korea. Also, several research results are reported on the examination of the roles of inserted pipes and grouted materials in UAM. But, because of its empirical design and construction methodology, more qualitative and systematic design sequences are needed. Therefore, above sequences using numerical analysis are proposed and, the effects of some design parameters were studied in this research. In order to acco,mplish these objects, first, the roles of pipe and grouting materials, steel-rib and the others in ground improving mechanism of UAM are clarified. Second, the effects of design parameters are investigated through parametric studies. Design parameters are as follows; 1) ground condition, 2) overburden, 3) geometrical formulation of pipes, 4) grouting region and 5) characteristics of pipes.

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The Response of a Single Pile and Pile Groups to Tunnelling Performed in Weathered Rock (풍화암에서 실시된 터널굴착으로 인한 단독말뚝 및 군말뚝의 거동)

  • Lee, Cheol Ju
    • KSCE Journal of Civil and Environmental Engineering Research
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    • v.32 no.5C
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    • pp.199-210
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    • 2012
  • The effects of tunnelling in weak weathered rock on the behaviour of a pre-existing single pile and pile groups ($3{\times}3$ and $5{\times}5$ pile groups) above a tunnel have been studied by carrying out three-dimensional (3D) elasto-plastic numerical analyses. Numerical modelling of such effects considers the response of the single pile and pile groups in terms of tunnelling-induced ground and pile settlement as well as changes of the shear transfer mechanism at the pile-soil interface due to tunnelling. Due to changes in the relative shear displacement between the pile and the soil at the pile-soil interface with tunnel advancement, the shear stresses and axial pile force distributions along the pile change drastically. Based on the computed results, upward shear stresses are induced up to about Z/L=0.775 from the pile top, while downward shear stresses are mobilised below Z/L=0.775, resulting in a reduction in the axial pile force distribution with depth equivalent to a net increase in the tensile force on the pile. A maximum tensile force of about $0.36P_a$ developed on the single pile solely due to tunnelling, where $P_a$ is the service axial pile loading prior to tunnelling. The degree of interface shear strength mobilisation at the pile-soil interface was found to be a key factor governing pile-soil-tunnelling interaction. Overall it has been found that the larger the number of piles, the greater is the effect of tunnelling on the piles in terms of pile settlement, while changes of the axial pile forces for the piles in the groups are smaller than for a single pile due to the shielding effect. The reduction of apparent allowable pile capacity due to tunnelling-induced pile head settlement was significant, in particular for piles inside the groups.

Molecular Biological Study of Anti-cancer Effects of Bee Venom on Human Melanoma Cell (약침용봉독액(藥鍼用蜂毒液)이 흑색종세포(黑色腫細胞)에 미치는 항암효과(抗癌效果)에 대(對)한 분자생물학적(分子生物學的) 연구(硏究))

  • Park, Chan-Yol;Nam, Sang-Soo;Kim, Chang-Hwan;Lee, Jae-Dong;Kang, Sung-Keel;Lee, Yun-Ho;Ahn, Byoung-Choul
    • Journal of Acupuncture Research
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    • v.17 no.2
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    • pp.169-186
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    • 2000
  • To study anti-cancer effect and molecular biological mechanism of bee venom for aqua-acupuncture, the effects of bee venom on cell viability, apoptosis, and cell cycle were analyzed using MTT assay, tryphan blue assay, [3H]thymidine release assay, flow cytometric analysis, activity of caspase-3 protease activity assay, and immunocytometric analysis of PCNA. To explore whether anti-cancer effects of bee venom are associated with the transcriptional control of gene expression, quantitative RT-PCR analysis of apoptosis- and cell cycle-related genes was performed. The obtained results are summarized as follows: 1. The MTT assay demonstrated that cell viability was decreased by bee venom in a dose-dependant manner. 2. Significant induction of apoptosis was identified using tryphan blue assay, [$^3H$]thymidine release assay, and flow cytometric analysis of sub $G_1$ fraction. 3. In analysis of caspase-3 protease activity, the activity had increased significantly, in a dose-dependant manner. 4. Quantitative RT-PCR analysis of the apoptosis-related genes showed that Bcl-2 and $Bcl-X_L$ were down-regulated whereas Bax was up-regulated by bee venom treatment. 5. In flow cytometric analysis of cell cycle and immunocytometric analysis of PCNA expression, cell numbers of $G_1$ phase was increased by a dose-dependant manner. 6. In quantitative RT-PCR analysis of the cell cycle-related genes, p21, p27, and p57 were increased, while Cyclin D1, CDK4, c-Myc, c-Fos, and Histone H3 were decreased. In contrast, there were no remarkable changes in expression levels of CDC2 and c-Jun.

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Hydrochemistry, Isotopic Characteristics, and Formation Model Geothermal Waters in Dongrae, Busan, South Korea (부산 동래 온천수의 수리화학 및 동위원소 특성, 생성모델 연구)

  • Yujin Lee;Chanho Jeong;Yongcheon Lee
    • The Journal of Engineering Geology
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    • v.34 no.2
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    • pp.229-248
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    • 2024
  • This investigated the hydrogeochemical and isotopic characteristics of geothermal waters, groundwaters, and surface waters in Dongrae-gu, Busan, South Korea, in order to determine the origins of the salinity components in the geothermal waters, and their formation mechanisms and heat sources The geothermal waters are Na-Cl-type, distinct from surrounding groundwaters (Na-HCO3- and, Ca-HCO3- (SO4, Cl)-type) and surface waters (Ca-HCO3(SO4, Cl)-type). This indicates the geothermal waters formed at depth as compared with the groundwaters. δ18O and δD values of the geothermal waters are relatively depleted as compared with the groundwaters, due to altitude effects and deep circulation of the geothermal waters. Helium and neon isotope ratios (3 He/4He and, 4He/20Ne) of the geothermal waters plot on a single mixing line between mantle (3He = 3.76~4.01%) and crust (4He = 95.99~96.24 %), indirectly suggesting that the heat source is due to the decay of radioactive elements in rocks. The geothermal reservoir temperatures were calculated using the silica-enthalpy and Giggenbach models, yielding values of 82~130℃, and the depth of the geothermal reservoir is estimated to be 1.7~2.9 km below the surface. The correlation between Cl/Na and Cl/HCO3 for the Dongrae geothermal waters requires the input of salty water. The supply of saline composition is interpreted due to the dissolution of residual paleo-seawater.

Analysis of Childhood Rapidly Progressive Glomerulonephritis (소아 급속 진행성 사구체신염의 임상적 고찰)

  • Uhm Ji Hyun;Kim Mi Jin;Lee Young-Mock;Kim Ji Hong;Lee Jae Seung;Kim Pyung-Kil;Hong Soon Won;Jeung Hyeun Joo
    • Childhood Kidney Diseases
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    • v.5 no.2
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    • pp.78-86
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    • 2001
  • Purpose: Rapidly progressive glomerulonephritis (RPGN) is characterized by the rapid increase in serum creatitnin and crescents formation involving more than $50\%$ of glomeruli. 10 patients who had been treated for RPGN were studied retrospectively for thier underlying diseases and clinical features Method: Cilinical review was performed on 10 children who were diagnosed with RPGN by clinical features and renal biopsy and followed up at department of pediatrics during tile last 10 years, from May 1990 to May 2000. Result: There were 6 males and 4 females between the ages of 2.1 and 14.3 years (mean $10.9{\pm}3.8$). 3 had Henoch-$Sch{\ddot{o}}nlein$ purpura nephritis; 2, idiopathic rapidly progressive glomerulonephritis; 2, lupus nephritis; 1, hemolytic uremic syndrome; 1, membranous glomerulonephritis and 1, microscopic polyangiitis. The most common chief complaints were gross hematuria and oliguria. Initial clinical features included proteinuria, edema, hypertension, nausea and arthralgia. Mean serum BUN was $74.2{\pm}39.1\;mg/dL$ mean serum creatinin, $3.2{\pm}1.8\;mg/dL$ and mean creatinin clearance, $26.5{\pm}13.2\;mL/min/1.73m^2$. Antineutrophil cytoplasmic antibody was positive only in microscopic polyangiitis. ANA and Anti-DNA antibody were positive in two lupus nephritis patients. Serum complements were decreased in 4 patients. All patients except Hemolytic uremic syndrome received steroid pulse therapy and immunosupressive agents. 3 patients were performed acute peritoneal dialysis and 2 patients were given plasmapheresis. At the last follow up, 1 patient was dead, 4 patients had elevated serum creatinin, 2 of these 4 patients were on chronic ambulatory peritoneal dialysis and 6 patients had normal renal function. Conclusion: Rapidly progressive glomerulonephritis is a medical emergency that requires very rapid diagnosis, classification, and therapy. Appropriate therapy selected on the basis of underlying disease mechanism can substantially improve renal survival. (J. Korean Soc Pediatr Nephrol 2001 ; 5 : 78-86)

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Molecular Miology of the Poliovirus (폴리오바이러스의 분자생물학)

  • 최원상
    • Journal of Life Science
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    • v.7 no.4
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    • pp.392-401
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    • 1997
  • The poliovirus is a small, and non-enveloped virus. The RNA genome of poliovirus is continuous, linear, and has a single open reading frame. This polyprotein precursor is cleaved proteolytically to yield mature products. Most of the cleavages occur by viral protease. The mature proteins derived from the P1 polyprotein precursor are the structural components of the viral capsid. The initial cleavage by 2A protease is indirectly involved in the cleavage of a cellular protein p220, a subunit of the eukaryotic translation initiation factor 4F. This cleavage leads to the shut-off of cap-dependent host cell translation, and allows poliovirus to utilize the host cell machinery exclusively for translation its own RNA, which is initiated by internal ribosome entry via a cap-independent mechanism. The functional role of the 2B, 2C and 2BC proteins are not much known. 2B, 2C, 2BC and 3CD proteins are involved in the replication complex of virus induced vesicles. All newly synthesized viral RNAs are linked with VPg. VPg is a 22 amino acid polypeptide which is derived from 3AB. The 3C and 3CD are protease and process most of the cleavage sites of the polyprotein precursor. The 3C protein is also involved in inhibition of RNA polymerase II and III mediated transcription by converting host transcription factor to an inactive form. The 3D is the RNA dependent RNA polymerase. It is known that poliovirus replication follows the general pattern of positive strand RNA virus. Plus strand RNA is transcribed into complementary minus strand RNA that, in turn, is transcribed for the synthesis of plus strand RNA is transcribed into complementary minus strand RNA that, in turn, is transcribed for the synthesis of plus strand RNA strands. Poliovirus RNA synthesis occurs in a membranous environment but how the template RNA and proteins required for RNA replication assemble in the membrane is not much known. The RNA requirements for the encapsidation of the poliovirus genome (packaging signal) are totally unknown. The poliovirus infection cycle lasts approximately 6 hours.

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