• Title/Summary/Keyword: L5 signal

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Paw Edema was Reduced in Carrageenan Induced Acute Inflammation in Stat4 Deficient Mice

  • Zheng, Long-Tai;Baik, Haing-Woon;Lee, Seong-Kyu;Cho, Jeong-Je;Park, Cheung-Seog;Hong, Mee-Suk;Chung, Joo-Ho;Yim, Sung-Vin
    • Molecular & Cellular Toxicology
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    • v.2 no.4
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    • pp.257-261
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    • 2006
  • Signal transducer and activator of transcription 4 (STAT4) is one of the important mediators in generating inflammation and immune responses. To address the role of Stat4 in carrageenan induced acute inflammation, we performed paw edema measurement and 7.4 k mouse cDNA microarray analysis in carrageenan induced acute inflammation in Stat4 knockout (-/-) mice. Male BALB/c (n=8) and Stat4 -/- (n=5) were used and paw edema was induced with injection of $30\;{\mu}L$ of 1% carrageenan into plantar surface of right hind paw. Next, we isolated the mRNA in mouse whole brain and analyzed cDNA microarray profiles for the changes of the brain expression in Stat4 -/- mice. Interestingly, the increase in paw volume of Stat4 -/- mice was reduced by about 30% as compared to that of wild type. The cDNA microarray analysis revealed the altered expressions of several cytokines (Tnf, Il6, and Il4) and pain-associated proteins (Ptgs2, Gabra6, and Gabbr1) in Stat4 -/- mice. Our results suggest that Stat4 may be related to the inhibitory responses on carrageenan induced acute inflammation.

Effects of Saturated Long-chain Fatty Acid on mRNA Expression of Genes Associated with Milk Fat and Protein Biosynthesis in Bovine Mammary Epithelial Cells

  • Qi, Lizhi;Yan, Sumei;Sheng, Ran;Zhao, Yanli;Guo, Xiaoyu
    • Asian-Australasian Journal of Animal Sciences
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    • v.27 no.3
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    • pp.414-421
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    • 2014
  • This study was conducted to determine the effects of saturated long-chain fatty acids (LCFA) on cell proliferation and triacylglycerol (TAG) content, as well as mRNA expression of ${\alpha}s1$-casein (CSN1S1) and genes associated with lipid and protein synthesis in bovine mammary epithelial cells (BMECs). Primary cells were isolated from the mammary glands of Holstein dairy cows, and were passaged twice. Then cells were cultured with different levels of palmitate or stearate (0, 200, 300, 400, 500, and 600 ${\mu}M$) for 48 h and fetal bovine serum in the culture solution was replaced with fatty acid-free BSA (1 g/L). The results showed that cell proliferation tended to be increased quadratically with increasing addition of stearate. Treatments with palmitate or stearate induced an increase in TAG contents at 0 to 600 ${\mu}M$ in a concentration-dependent manner, and the addition of 600 ${\mu}M$ was less effective in improving TAG accumulation. The expression of acetyl-coenzyme A carboxylase alpha, fatty acid synthase and fatty acid-binding protein 3 was inhibited when palmitate or stearate were added in culture medium, whereas cluster of differentiation 36 and CSN1S1 mRNA abundance was increased in a concentration-dependent manner. The mRNA expressions of peroxisome proliferator-activated receptor gamma, mammalian target of rapamycin and signal transducer and activator of transcription 5 with palmitate or stearate had no significant differences relative to the control. These results implied that certain concentrations of saturated LCFA could stimulate cell proliferation and the accumulation of TAG, whereas a reduction may occur with the addition of an overdose of saturated LCFA. Saturated LCFA could up-regulate CSN1S1 mRNA abundance, but further studies are necessary to elucidate the mechanism for regulating milk fat and protein synthesis.

High-Level Secretory Expression of Human Procarboxypeptidase B by Fed-Batch Cultivation of Pichia pastoris and its Partial Characterization

  • Kim, Mi-Jin;Kim, Sang-Hyuk;Lee, Jae-Hyung;Seo, Jin-Ho;Lee, Jong-Hwan;Kim, Jong-Hyun;Kim, Yeon-Hee;Nam, Soo-Wan
    • Journal of Microbiology and Biotechnology
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    • v.18 no.12
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    • pp.1938-1944
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    • 2008
  • The procpb gene encoding human procarboxypeptidase B (proCPB, GeneBank access code AJ224866) was cloned and its Pichia expression plasmid, $pPIC9{\alpha}$/hproCPB (9.2 kb), was constructed, in which procpb was under the control of the AOXl promoter and connected to the downstream of the mating factor ${\alpha}$-1 ($MF{\alpha}1$) signal sequence. The plasmid was linearized by digestion with Sacl, and integrated into the genome of P. pastoris strain GS115. By culturing of Pichia transformant on methanol medium, the human proCPB was successfully expressed and secreted into the culture supernatant. Moreover, Western blot analysis of the extracellular proteins showed proCPB bands clearly at a molecular mass of 45 kDa, confirming the expression of proCPB with its right size. The CPB activity reached about 3.5 U/ml and 12.7 U/ml in the flask and fermentor batch cultures of Pichia transformant, respectively. No CPB enzyme activity was found in the intracellular fraction. When the fed-batch cultivation was performed with methanol and glycerol mixture as a feeding medium, the extracellular CPB activity was increased to 42.0 U/ml, which corresponds to a 3.3-fold higher level of CPB activity than that of batch culture. The $K_m$ and $k_{cat}$ values of recombinant human CPB enzyme for hippuryl-$_L$-Arg as a substrate were estimated to be 0.16 mM and $11.93\;sec^{-1}$, respectively.

Decreased Expression of Phospholipase C-$\beta$2 in Human Platelets with Impaired Function

  • Lee, Sang-Bong;A. Koneti Rao;Lee, Kweon-Haeng;Xu Yang;Bae, Yun-Soo
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1996.11a
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    • pp.75-84
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    • 1996
  • Platelets from a patient with a mild inherited bleeding disorder and abnormal platelet aggregation and secretion show reduced generation of inositol 1,4,5-trisphosphate (IP$_3$), mobilization of intracellular Ca$\^$2+/, and phosphorylation of pleckstrin in response to several G protein mediated agonists, suggesting a possible defect at the level of phospholipase C (PLC) activation. A procedure was developed that allows quantitation of platelet PLC isozymes. After fractionation of platelet extracts by high-performance liquid chromatography, seven, out often known PLC isoforms were detected by immunoblot analysis. The amount of these isoforms in normal platelets decreased in the order PLC-${\gamma}$2 > PLC-${\beta}$2 > PLC-${\beta}$3 > PLC-${\beta}$l > PLC-${\gamma}$ > PLC-$\delta$1 > PLC-${\beta}$4. Compared with normal platelets, platelets from the patient contained approximately one-third the amount of PLC-${\beta}$2, whereas PLC-${\beta}$4 was increased threefold. These results suggest that the impaired platelet function in the patient in response to multiple G protein mediated agonists is attributable to a deficiency of PLC-${\beta}$2. They document for the first time a specific PLC isozyme deficiency in human platelets and provide an unique opportunity to understand the role of different PLC isozymes in normal platelet function.

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The Kwinana Shoreline Fumigation Experiment in Western Australia, Australia

  • Yoon, I.H.;Sawford, B.L;Manins, P.C.
    • Proceedings of the Korean Environmental Sciences Society Conference
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    • 1996.04a
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    • pp.22-22
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    • 1996
  • ;The Kwinana Shoreline Fumigation Experiment(KSFE) took place in Fremantle, WA, Australia between 23 January and 8 February, 1995. All measurement systems performed to expectation. The CSIRO DAR(Division of Atmospheric Research) LIDAR measured plume sections from near the Kwinana Power Station(KPS) stacks to up to about 5 km downstream. It also measured boundary layer aerosols and the structure of the boundary layer on some occasions. Both stages A and C of KPS were used as tracers at different times. Radiosonde and double theodolite sounding systems measured temperature, humidity, air pressure and wind structure at the coast(Woodman Point) and at the inland(ALCOA residue dump) site at intervals of roughly two hours. These were supplemented by mid afternoon soundings(radiosonde and single theodolite) by Department of Environmental Protection(DEP) at Swanbourne. The Flinders aircraft measured wind, turbulence and temperature structure of the atmospheric boundary layer, concentrations of $C0_2,\;0_3,\;S0_2\;and\;NO_x$ in the smoke plumes and surface radiation over both land and sea. CSIRO DCET(Division of Coal and Energy Technology) vehicle successfully interceptde many smoke plumes and using a range of tracers will be able to identify the various sources much of the time. Routine data from the DEP and Kwinana Industrial Council(KIC) air quality monitoring networks were also automatically logged. Murdoch University measured surface heat flux at Hope Valldy monitoring station and also at Wattleup monitoring station for the last five days. The heart of the LIDAR system is a Neodymium-doped Yttrium-aluminumgarnet(Nd:Y AG) laser operating at a fundamental wavelength of 1064 nm, with harmonics fo 532 nm and 355 nm. A small fraction of the laser beam is scattered back to the LIDAR, collected by a telescope and detedted by a photomultiplier tube. The intensity of the signal as a function of time is a measure of the particle concentration as a function of distance along the line of the laser shot. The results of nine days special field observations are summarized in detail.etail.

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Inhibition of Cyclooxygenase and Prostaglandin E2 Synthesis by Crude Methanolic Extract from Euonymus Alatus (Thunb.) Sieb in SKBR3 Human Breast Cancer Cell Line

  • Kim Joong-Oh;Jang Tae-Hyun;Kim Min-Sung;Kim Dong-Il;Lee Tae-Kyun
    • The Journal of Korean Medicine
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    • v.26 no.1 s.61
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    • pp.37-45
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    • 2005
  • In the present study, we examined the effect of crude methanolic extract (CME) from Euonymus alatus (Thunb.) Sieb on arachidonic acid (AA) cascade in SKBR3 human breast cancer cell line. CME had a potent inhibitory activity of prostaglandin E2 (PGE2) release induced by A23187, a $Ca^{2+}$ ionophore. The inhibition was concentration-dependent, with the 50 value of about 5 M. CME had no inhibitory effect on A23187-induced phosphorylation of p42/p44 extracellular signal regulated kinase/mitogen-activated protein kinase or on the liberation of [14C]-AA from the cells labeled with [14C]-AA. However, CME concentration-dependently inhibited the conversion of AA to $PGE_2$ in microsomal preparations, showing its possible inhibition of cyclooxygenase (COX). In enzyme assay in vitro, CME inhibited the activities of both constitutive COX (COX­I) and inducible COX (COX-2) in a concentration-dependent manner, with the 50 values of about 0.8 and 2M, respectively. Lineweaver-Burk plot analysis indicated that CME competitively inhibited the activities of both COX-l and -2. This study is a first demonstration that CME directly inhibits COX activity.

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Cloning and Expression of a Thermostable ${\alpha}$-Galactosidase from the Thermophilic Fungus Talaromyces emersonii in the Methylotrophic Yeast Pichia pastoris

  • Simila, Janika;Gernig, Anita;Murray, Patrick;Fernandes, Sara;Tuohy, Maria G.
    • Journal of Microbiology and Biotechnology
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    • v.20 no.12
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    • pp.1653-1663
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    • 2010
  • The first gene (${\alpha}$-gal1) encoding an extracellular ${\alpha}$-Dgalactosidase from the thermophilic fungus Talaromyces emersonii was cloned and characterized. The ${\alpha}$-gal1 gene consisted of an open reading frame of 1,792 base pairs interrupted by six introns that encoded a mature protein of 452 amino acids, including a 24 amino acid secretory signal sequence. The translated protein had highest identity with other fungal ${\alpha}$-galactosidases belonging to glycosyl hydrolase family 27. The ${\alpha}$-gal1 gene was overexpressed as a secretory protein with an N-terminal histidine tag in the methylotrophic yeast Pichia pastoris. Recombinant ${\alpha}$-Gal1 was secreted into the culture medium as a monomeric glycoprotein with a maximal yield of 10.75 mg/l and purified to homogeneity using Hisbinding nickel-agarose affinity chromatography. The purified enzyme was maximally active at $70^{\circ}C$, pH 4.5, and lost no activity over 10 days at $50^{\circ}C$. ${\alpha}$-Gal1 followed Michaelis-Menten kinetics ($V_{max}\;of\;240.3{\mu}M/min/mg,\;K_m\;of\;0.294 mM$) and was inhibited competitively by galactose ($K_m{^{obs}}$ of 0.57 mM, $K_i$ of 2.77 mM). The recombinant T. emersonii ${\alpha}$-galactosidase displayed broad substrate preference, being active on both oligo- and polymeric substrates, yet had strict specificity for the ${\alpha}$-galactosidic linkage. Owing to its substrate preference and noteworthy stability, ${\alpha}$-Gal1 is of particular interest for possible biotechnological applications involving the processing of plant materials.

Effects of fermented black ginseng on wound healing mediated by angiogenesis through the mitogen-activated protein kinase pathway in human umbilical vein endothelial cells

  • Park, Jun Yeon;Lee, Dong-Soo;Kim, Chang-Eop;Shin, Myoung-Sook;Seo, Chang-Seob;Shin, Hyeun-Kyoo;Hwang, Gwi Seo;An, Jun Min;Kim, Su-Nam;Kang, Ki Sung
    • Journal of Ginseng Research
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    • v.42 no.4
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    • pp.524-531
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    • 2018
  • Background: Fermented black ginseng (FBG) is produced through several cycles of steam treatment of raw ginseng, at which point its color turns black. During this process, the original ginsenoside components of raw ginseng (e.g., Re, Rg1, Rb1, Rc, and Rb2) are altered, and less-polar ginsenosides are generated (e.g., Rg3, Rg5, Rk1, and Rh4). The aim of this study was to determine the effect of FBG on wound healing. Methods: The effects of FBG on tube formation and on scratch wound healing were measured using human umbilical vein endothelial cells (HUVECs) and HaCaT cells, respectively. Protein phosphorylation of mitogen-activated protein kinase was evaluated via Western blotting. Finally, the wound-healing effects of FBG were assessed using an experimental cutaneous wounds model in mice. Results and Conclusion: The results showed that FBG enhanced the tube formation in HUVECs and migration in HaCaT cells. Western blot analysis revealed that FBG stimulated the phosphorylation of p38 and extracellular signal-regulated kinase in HaCaT cells. Moreover, mice treated with $25{\mu}g/mL$ of FBG exhibited faster wound closure than the control mice did in the experimental cutaneous wounds model in mice.

The Immediate Effect of Electroacupuncture at the B62(Shinmaek) K6(Chohae) on the EEG of Vascular Dementia (신맥 조해의 전침자극이 치매환자의 뇌파에 미치는 영향)

  • Park, Woo-Soon;Lee, Tae-Young;Kim, Soo-Yong;Lee, Kwang-Gyu;Yuk, Sang-Won;Lee, Chang-Hyun;Lee, Sang-Ryong
    • Journal of Acupuncture Research
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    • v.18 no.2
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    • pp.67-78
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    • 2001
  • The aim of this study was to examine the effects of low frequency electroacupuncture(EA) at the $B_{62}$ (Shinmaek) $K_6$(Chohae) on vascular dementia in humans using nonlinear dynamics. Electroencephalogram(EEG) is a multi-scaled signal consisting of several components of time series with different dominant frequency ranges and different origins. Nonlinear measures of the EEG like the correlation dimension ($D_2$) and the first positive Lyapunov exponent ($L_1$) reflect the complexity of the EEG. In this study, $D_2$ was used as a measure of complexity. Sixteen channel EEG study was carried out in six subjects (5 females and 1 males; $age=83.83{\pm}7.19years$). We found that the baseline $D_2$ values of the EEG at F4 and F8 channels (P<0.01) were lowered than during the acupuncture treatment, indicating decreased complexity of the EEG. However, the comparison with that before and after the treatment shows no significant differences in all channels.

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A Study on the Partial Discharge Phase Properties with Branch Type Eleotrical Tree Growth in XLPE Cable Insulation (XLPE 케이블 절연체에서의 가지형 전기트리 성장에 따른 부분방전 위상 특성 연구)

  • Gang, Dong-Sik;Seon, Jong-Ho;Kim, Wi-Yeong;Lee, Hong-Sik;Park, Jeong-Hu
    • The Transactions of the Korean Institute of Electrical Engineers C
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    • v.51 no.5
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    • pp.213-221
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    • 2002
  • In order to improve the reliability of XLPE cables, it is necessary to find the useful diagnostic parameter according to long term aging. This paper described the change of partial discharge(PD) phase Properties of XLPE cable insulation with branch type electrical tree degradation. for long term aging. To understand the PD Properties with $\phi$ -q-n distributed shape of XLPE insulation, specimens were prepared by 22.9㎸ distribution cable and made in a type of block(16${\times}$16${\times}$3[mm]). Ogura needles having tip radius of l0${\mu}{\textrm}{m}$ were inserted into each block pieces. The measuring system was consisted of PD detector, digitizer for digital conversion, VXI system for signal processing. The PD properties of the specimens were measured from initiation of tree to breakdown and their characteristics were analyzed. We analyzed the relationship between electrical properties(PD Quantity, PD initiation angle, PD extinction angle, PD occurrence angle : (PD extinction angle - PD initiation angle)) and the normalized aging rate. We found PD parameter, PD initiation angle and occurrence angle, which are a useful diagnostic parameter in estimating the branch type electrical tree for XLPE insulation condition.