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검색결과 492건 처리시간 0.023초

Heat Shock Protein $90{\beta}$ Inhibits Phospholipase $C{\gamma}-1$ Activity in vitro

  • ;;장종수
    • 대한의생명과학회지
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    • 제12권4호
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    • pp.419-425
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    • 2006
  • Phospholipase $C-{\gamma}1\;(PLC-{\gamma}1)$ is an important signaling molecule for cell proliferation and differentiation. $PLC-{\gamma}1$ contains two pleckstrin homology (PH) domains, which are responsible for protein-protein interaction and protein-lipid interaction. $PLC-{\gamma}1$ also has two Src homology (SH)2 domains and a SH3 domain, which are responsible for protein- protein interaction. To identity proteins that specifically binds to PH domain of $PLC-{\gamma}1$, we prepared and incubated the glutathione S-transferase(GST)-fused PH domains of $PLC-{\gamma}1$ with COS7 cell lysate. We found that 90 kDa protein specifically binds to PH domain of $PLC-{\gamma}1$. By matrix-assisted laser desorption ionization time of flight-mass spectrometry, the 90 kDa protein revealed to be heat shock protein (Hsp) $90{\beta}$. Hsp $90{\beta}$ is a molecular chaperone that stabilizes and facilitates the folding of proteins that are involved in cell signaling, including receptors for steroids hormones and a variety of protein kinases. To know whether Hsp $90{\beta}$ affects on $PLC-{\gamma}1$ activity, we performed $PIP_2$ hydrolyzing activity of $PLC-{\gamma}1$ in the presence of purified Hsp $90{\beta}$ in vitro. Our results show that the Hsp $90{\beta}$ dose-dependently inhibits the enzymatic activity of $PLC-{\gamma}1$ and further suggest that Hsp $90{\beta}$ regulates cell growth and differentiation via regulation of $PLC-{\gamma}1$ activity.

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RGS Protein Specificity Towards Gq- and Gi/o-Mediated ERK 1/2 and Akt Activation, in vitro

  • Anger, Thomas;Klintworth, Nils;Stumpf, Christian;Daniel, Werner G.;Mende, Ulrike;Garlichs, Christoph D.
    • BMB Reports
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    • 제40권6호
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    • pp.899-910
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    • 2007
  • Extracellular Regulated Kinases (ERK) and Protein Kinase B (Akt) are intermediaries in relaying extracellular growth signals to intracellular targets. Each pathway can become activated upon stimulation of G protein-coupled receptors mediated by $G_q$ and $G_{i/o}$ proteins subjected to regulation by RGS proteins. The goal of the study was to delineate the specificity in which cardiac RGS proteins modulate $G_{q^-}$ and $G_{i/o}$-induced ERK and Akt phosphorylation. To isolate $G_{q^-}$ and $G_{i/o}$-mediated effects, we exclusively expressed muscarinic $M_2$ or $M_3$ receptors in COS-7 cells. Western blot analyses demonstrated increase of phosphorylation of ERK 1.7-/3.3-fold and Akt 2.4-/6-fold in $M_{2^-}/M_{3^-}$ expressing cells through carbachol stimulation. In co-expressions, $M_3/G_q$-induced activation of Akt was exclusively blunted through RGS3s/RGS3, whereas activation of ERK was inhibited additionally through RGS2/RGS5. $M_2/G_{i/o}$ induced Akt activation was inhibited by all RGS proteins tested. RGS2 had no effect on $M_2/G_{i/o}$-induced ERK activation. The high degree of specificity in RGS proteins-depending modulation of $G_{q^-}$ and $G_{i/o}$-mediated ERK and Akt activation in the muscarinic network cannot merely be attributed exclusively to RGS protein selectivity towards $G_q$ or $G_{i/o}$ proteins. Counter-regulatory mechanisms and inter-signaling cross-talk may alter the sensitivity of GPCR-induced ERK and Akt activation to RGS protein regulation.

Identification of a Novel Human Lysophosphatidic Acid Acyltransferase, LPAAT-theta, Which Activates mTOR Pathway

  • Tang, Wenwen;Yuan, Jian;Chen, Xinya;Gu, Xiuting;Luo, Kuntian;Li, Jie;Wan, Bo;Wang, Yingli;Yu, Long
    • BMB Reports
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    • 제39권5호
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    • pp.626-635
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    • 2006
  • Lysophosphatidic acid acyltransferase (LPAAT) is an intrinsic membrane protein that catalyzes the synthesis of phosphatidic acid (PA) from lysophosphatidic acid (LPA). It is well known that LPAAT is involved in lipid biosynthesis, while its role in tumour progression has been of emerging interest in the last few years. To date, seven members of the LPAAT gene family have been found in human. Here we report a novel LPAAT member, designated as LPAAT-theta, which was 2728 base pairs in length and contained an open reading frame (ORF) encoding 434 amino acids. The LPAAT-theta gene consisted of 12 exons and 11 introns, and mapped to chromosome 4q21.23. LPAAT-theta was ubiquitously expressed in 18 human tissues by RT-PCR analysis. Subcellular localization of LPAAT-theta-EGFP fusion protein revealed that LPAAT-theta was distributed primarily in the endoplasmic reticulum (ER) of COS-7 cells. Furthermore, we found that the overexpression of LPAAT-theta can induce mTOR-dependent p70S6K phosphorylation on Thr389 and 4EBP1 phosphorylation on Ser65 in HEK293T cells.

Efficacy of a DNA Vaccine Carrying Eimeria maxima Gam56 Antigen Gene against Coccidiosis in Chickens

  • Xu, Jinjun;Zhang, Yan;Tao, Jianping
    • Parasites, Hosts and Diseases
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    • 제51권2호
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    • pp.147-154
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    • 2013
  • To control coccidiosis without using prophylactic medications, a DNA vaccine targeting the gametophyte antigen Gam56 from Eimeria maxima in chickens was constructed, and the immunogenicity and protective effects were evaluated. The ORF of Gam56 gene was cloned into an eukaryotic expression vector pcDNA3.1(zeo)+. Expression of Gam56 protein in COS-7 cells transfected with recombinant plasmid pcDNA-Gam56 was confirmed by indirect immunofluorescence assay. The DNA vaccine was injected intramuscularly to yellow feathered broilers of 1-week old at 3 dosages (25, 50, and $100{\mu}g/chick$). Injection was repeated once 1 week later. One week after the second injection, birds were challenged orally with $5{\times}10^4$ sporulated oocysts of E. maxima, then weighed and killed at day 8 post challenge. Blood samples were collected and examined for specific peripheral blood lymphocyte proliferation activity and serum antibody levels. Compared with control groups, the administration of pcDNA-Gam56 vaccine markedly increased the lymphocyte proliferation activity (P<0.05) at day 7 and 14 after the first immunization. The level of lymphocyte proliferation started to decrease on day 21 after the first immunization. A similar trend was seen in specific antibody levels. Among the 3 pcDNA-Gam56 immunized groups, the median dosage group displayed the highest lymphocyte proliferation and antibody levels (P<0.05). The median dosage group had the greatest relative body weight gain (89.7%), and the greatest oocyst shedding reduction (53.7%). These results indicate that median dosage of DNA vaccine had good immunogenicity and immune protection effects, and may be used in field applications for coccidiosis control.

Lack of Any Relationship between ABO and Rh Blood Groups and Clinicopathological Features in Patients with Gastrointestinal Stromal Tumors: Turkish Oncology Group

  • Urun, Yuksel;Utkan, Gungor;Yalcin, Suayib;CosKun, Hasan Senol;Kocer, Murat;Ozdemir, Nuriye Yildirim;Kaplan, Mehmet Ali;Arslan, Ulku Yalcintas;Ozdemir, Feyyaz;Oztuna, Derya;Akbulut, Hakan;Icli, Fikri
    • Asian Pacific Journal of Cancer Prevention
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    • 제13권8호
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    • pp.4129-4131
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    • 2012
  • Background: An association between the ABO blood group and the risk of certain malignancies, including pancreatic and gastric cancer, has been reported previously. However, it is unclear whether this association is valid for gastrointestinal stromal tumors (GIST). In this study, ABO blood groups and the Rh factor were investigated in a series of GIST cases. Material and Methods: In 162 patients with GIST, blood group and Rh factor were examined and compared with a control group of 3,022,883 healthy volunteer blood donors of the Turkish Red Crescent between 2004 and 2011. The relationship of blood groups with tumor size, mitotic activity, and age were also evaluated. Results: Overall, the ABO blood group and Rh factor distributions of the 162 patients with GIST were similar to those of the general population. There were no significant differences between both ABO blood types and Rh factor in terms of tumor size, mitotic activity, and age. Conclusion: This is the first study reported on this issue. In our study, we didn't find any relationship between GIST and ABO blood group and Rh factor. However further studies with larger number of patients are needed to establish the role of blood groups in this population.

Expression and in vitro function of anti-cancer mAbs in transgenic Arabidopsis thaliana

  • Song, Ilchan;Kang, Yang Joo;Kim, Dae Heon;Kim, Mi Kyung;Ko, Kisung
    • BMB Reports
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    • 제53권4호
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    • pp.229-233
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    • 2020
  • The anti-colorectal cancer monoclonal antibody CO17-1A (mAb CO), which recognizes the tumor-associated antigen EpCAM, was expressed in transgenic Arabidopsis plants. PCR and western blot analyses showed the insertion and expression of heavy chain (HC)/HC fused to the KDEL ER retention modif (HCK) and light chain (LC) of mAb CO and mAb CO with HCK (mAb COK) in Arabidopsis transformants. Both plant-derived mAbP CO and mAbP COK were purified from a biomass of approximately 1,000 seedlings grown in a greenhouse. In sandwich ELISA, both mAbP CO showed a slightly higher binding affinity for the target, EpCAM, compared to mAbM CO. In cell ELISA, both mAbsP COs showed binding affinity to the human colorectal cancer cell line SW480. Furthermore, mAbM CO, mAbP CO, and mAbP COK exhibited dose and timedependent regression effects on SW480 cells in vitro. In summation, both mAbP CO and mAbP COK, expressed in Arabidopsis, recognized the target antigen EpCAM and showed anti-proliferative activity against human colorectal cancer cells.

The Characteristics and Change of Colors on Fashion Collections in 1990s

  • Kim, Honey;Kim, Young-In
    • International Journal of Costume and Fashion
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    • 제7권2호
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    • pp.18-31
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    • 2007
  • The purpose of this study is to examine color characteristics and color changes of the fashion collections through 1990s, and to provide the efficient color information for color planning upon fashion themes. For this research, a total of 30,084 colors were collected from Paris, Milan, London, New York Collections in 1990s. Those colors were first measured by the Pantone Textile Color Specifier and COS Color System and spectrophotometer(color eye 580). These measured color values $L^{\ast}a^{\ast}b^{\ast}$of CIE were converted into H V/C of Munsell System, and 12 tones of PCCS with 5 achromatic colors. The characteristics of collected colors were analyzed in general and by place, season and year. The results of the study are as follows : First, the hues of purple blue, yellow red, red, yellow and the tones of grayish, pale, white, black, dark grayish, dull, light grayish appeared mostly. Yellow was shown quite frequently in spring/summer while purple, purple blue, red and yellow red in fall/winter. White, pale, light, light grayish and light gray were shown more frequently in spring/summer while Black, dark grayish, grayish, dark gray and dark in fall/winter. Second, the characteristics of colors by 4 representative places were similar to the general characteristics of colors in 1990's. Third, There were distributed widely Red, Yellow Red, Yellow in the early 1990s, Green Yellow, Green, Blue Green in the mid of 1990s, and Purple Blue, Purple in the late of 1990s. The distribution range of chromatic colors showed wide in both of the early of 1990s and the mid of 1990s for a while, and achromatic colors of grayish, gray and black appeared mostly in the late of 1900s.

횡단방향 측선각도 변화에 따른 유량측정성과에 대한 실험 연구 (An experimental study on discharge measurements according to the change of traverse direction sideline angle)

  • 이재혁;석수원;심은증;김성훈;김태웅
    • 한국수자원학회:학술대회논문집
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    • 한국수자원학회 2011년도 학술발표회
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    • pp.330-334
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    • 2011
  • 자연하천의 유량값은 일반적으로 횡단면의 면적과 이를 통과하는 유속의 곱으로 산정된다. 유량측정 방법은 하천의 형태와 저 평수기 및 홍수기의 수위에 따라 도섭법, 교량법, 부자법 및 보트를 이용한 ADVM 측정법 등 다양한 방법으로 실시된다. 그러나 현장 여건에 따라 흐름에 직각이 아닌 사교에서 측정이 이루어지는 경우에는 단면적의 오차를 포함할 가능성이 크기 때문에 횡단면의 측선 각도에 따라 각보정을 실시해야 한다. 현재 사교에서 유량 측정을 실시하는 경우, 흐름의 직각을 기준으로 처짐각을 측량하여 각 측선에 $cos{\theta}$를 적용하여 단면적을 보정하고 있는데, 이 처짐의 정도가 유량의 참값에서 어느 정도의 영향을 미치는지에 대한 검토가 요구된다. 본 연구에서는 한강수계 왕숙천에 위치한 퇴계원 지점에서 실시간 수위에 따른 유속을 측정하였으며, 횡단면에 직각인 측선을 기준값으로 제시하고, 처짐각의 정도를 $10^{\circ}$, $20^{\circ}$, $30^{\circ}$, $40^{\circ}$, $50^{\circ}$까지 늘려 산정된 유량값을 기준값과 비교 분석하였다. 본 연구에 쓰인 측정기기는 Price AA 유속계이고, 측정방법으로는 유량의 흐름 방향을 기준하여 직각으로 수면에서 0.6d 지점의 유량측정방법(1점법)을 적용하였다. 그 결과 유량의 흐름 방향을 기준하여 직각인 경우 $1.39m^3/s$의 유량에서 보정 전 각 $10^{\circ}$의 유량 $1.36m^3/s$, $30^{\circ}$의 유량 $1.49m^3/s$, $50^{\circ}$의 유량 $2.25m^3/s$로 각이 클수록 단면적이 크게 나타나며 유량 역시도 과대 산정됨을 알 수 있었다. 따라서 도섭법을 이용한 유량측정이나 사교에서의 교량법 등을 적용하여 유량측정을 실시할 경우 유량의 흐름방향을 기준으로 직각으로 유량측정을 실시하여 유량을 산정하되 부득이한 경우로 사교에서의 측정이 이루어 졌을 시 흐름 방향을 기준으로 각도를 측정하여 크게 나타나는 단면적에 처짐각을 보정하여 유량을 산정함이 오차를 줄일 수 있으며, 신뢰성 있는 유량자료 생산의 방법이 라 할 수 있겠다.

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직관과 연결된 나선관 입구영역의 층류 유동 (Laminar Flow in the Entrance Region of Helical Tubes Connected with Straight Ones)

  • 김영인;박종호
    • 한국유체기계학회 논문집
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    • 제11권1호
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    • pp.9-17
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    • 2008
  • A numerical study for three-dimensional laminar flow in the entrance region of helical tubes connected with straight ones is carried out to investigate the effects of Reynolds number, pitch and curvature ratio on the oscillation periods of the flow. The fully elliptic governing equations were solved by means of a finite volume method. The fully developed laminar flow boundary condition was applied at the straight tube inlet. This results cover a curvature ratio range of 1/10${\sim}$1/320, a pitch range of 0.0${\sim}$3.2, and a Reynolds number range of 62.5${\sim}$2000. A comparison is made with previous experimental correlations and numerical data. The developments of velocity, local and average friction factors are discussed. The average friction factors are oscillatory in the entrance region of helical pipes. It has been found that the angle required for the flow to be similarly developed is most affected by the curvature ratio. The pitch and Reynolds number do not have any significant effect on the angle. The characteristic angle ${\phi}_c(={\phi}/sqrt{\delta})$, or the characteristic length to diameter ratio $s_c(=l\sqrt{\delta} cos(atan{\lambda})/d)$, can be useful to represent the development of flow in helical tubes. As the pitch increases and as the curvature ratio and Reynolds number decrease, the amplitude and the number of flow oscillations along the main streamwise direction decrease.

Cloning and Characterization of the IgA Fc Receptor from Swine

  • Chen, Yumei;Liu, Yunchao;Zhang, Gaiping;Feng, Hua;Ji, Pengchao;Wang, Guoqiang;Liu, Chang;Song, Yapeng;Su, Yunfang;Qiao, Songlin;Wang, Aiping
    • Journal of Microbiology and Biotechnology
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    • 제26권12호
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    • pp.2192-2198
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    • 2016
  • The myeloid-specific IgA Fc receptor ($Fc{\alpha}R$) is a cell surface molecule on immunocytes that provides a fundamental connection between humoral and cellular immunity. In this study, the full-length cDNA sequence of swine $Fc{\alpha}RI$ ($swFc{\alpha}RI$) was isolated and characterized and found to contain a 792-base-pair open reading frame, encoding a 264-amino-acid transmembrane glycoprotein with a predicted molecular mass of 29.4 kDa. The $swFc{\alpha}RI$ shares high amino acid sequence homology (>50%) with its counterparts from cattle, seal, and horse. Rosetting analysis confirmed that COS-7 cells transfected with an $swFc{\alpha}RI$ expression plasmid was able to combine with chicken erythrocytes sensitized with porcine IgA, but not IgG.