• Title/Summary/Keyword: 2-dimensional gel

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Comparison of Specific Proteins of Shiga Toxin-producing E. coli (STEC) Adhesion by Lactobacillus acidophilus Strains Using Two Dimensional Gel Electrophoresis (이차원 전기영동을 이용한 Lactobacillus acidophilus Strains의 Shiga Toxin-producing E. coli (STEC) 부착 억제와 관련된 단백질 발현 변화 분석)

  • Kim Young-Hoon;Moon Yong-Il
    • Food Science of Animal Resources
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    • v.26 no.2
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    • pp.263-268
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    • 2006
  • Probiotics including Lactobacillus acidophilus, refer to a group of nonpathogenic organisms that protect the human host against gastrointestinal(GI) infections by pathogenic bacteria such as Shiga toxin-producing E. coli(STEC). In the study, the inhibitory effects of STEC ATCC 43894 adhesion by L. acidophilus A4 was investigated on the HT-29 epithelial cells. Specific proteins regulated by cell Iysates of L. acidophilus A4 on STEC ATCC 43894 were also characterized by proteomic analysis. Both cell mass and Iysate of L. acidophilus A4 have exhibited the profound inhibitory activity on the HT-29 cells(about 1.5 log scale reduction). Two-dimensional gel electrophoresis(2-DE) revealed seven proteins that were up-regulated by cell Iysates of L. acidophilus A4 and three proteins that were down-regulated. In addition, three protein spots were only detected in the presence of cell Iysates. These results suggest that inhibitory effects of STEC adhesion by L. acidophilus may be due to the regulation of specific protein of STEC.

Impact of UV curing process on mechanical properties and dimensional accuracies of digital light processing 3D printed objects

  • Lee, Younghun;Lee, Sungho;Zhao, Xing Guan;Lee, Dongoh;Kim, Taemin;Jung, Hoeryong;Kim, Namsu
    • Smart Structures and Systems
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    • v.22 no.2
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    • pp.161-166
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    • 2018
  • In the last decade, there has been an exponential increase of scientific interest in smart additive manufacturing (AM) technology. Among the different AM techniques, one of the most commonly applied processes is digital light processing (DLP). DLP uses a digital projector screen to flash an ultraviolet light which cures photopolymer resins. The resin is cured to form a solid to produce parts with precise high dimensional accuracy. During the curing process, there are several process parameters that need to be optimized. Among these, the exposure time affects the quality of the 3D printed specimen such as mechanical strength and dimensional accuracy. This study examines optimal exposure times and their impact on printed part. It was found that there is optimal exposure time for printed part to have appropriate mechanical strength and accurate dimensions. The gel fraction and TGA test results confirmed that the improvement of mechanical properties with the increasing UV exposure time was due to the increase of crosslinked network formation with UV exposure time in acrylic resins. In addition, gel fraction and thermogravimetric analysis were employed to microscopically investigate how this process parameter impacts mechanical performance.

Fabricatoin and electrochemical property of 3-dimesional $LiMn_2O_4$ thin film (3차원 구조의 $LiMn_2O_4$ 박막전극의 제조 및 전기화학적 특성)

  • Park, Bo-Gun;Ryu, Jea-Hyeok;Park, Yong-Joon
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • 2008.06a
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    • pp.64-65
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    • 2008
  • 3D microbattery에 사용할 수 있는 $LiMn_2O_4$ 3차원 박막전극을 제조하여 그 전기화학적 특성을 관찰하였다. 3차원 구조의 형성을 위하여 먼저 polystyrene(PS) microsphere를 platinum이 증착된 Si/$SiO_2$ 기판위에 dip-coating 방식으로 코팅시켜 template로 사용하였다. 그 위에 sol-gel법을 이용, 박막을 형성시킨 후 template 를 제거하는 방식으로 $LiMn_2O_4$ 3차원 박막전극을 형성하였는데 이때 solution은 Lithium acetylacetonate[$LiCH_3CO-CHCOCH_3$], Manganese(III) acetylacetonate [Mn$(CH_3COCHCOCH_3)_3$]를 source 물질로 1-butanol과 acetic acid를 solvent로 활용하여 제조하였다.

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TNF-${\alpha}$ Up-regulated the Expression of HuR, a Prognostic Marker for Ovarian Cancer and Hu Syndrome, in BJAB Cells

  • Lee, Kyung-Yeol
    • IMMUNE NETWORK
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    • v.4 no.3
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    • pp.184-189
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    • 2004
  • Background: Hu syndrome, a neurological disorder, is characterized by the remote effect of small cell lung cancer on the neural degeneration. The suspicious effectors for this disease are anti-Hu autoantibodies or Hu-related CD8+ T lymphocytes. Interestingly, the same effectors have been suggested to act against tumor growth and this phenomenon may represent natural tumor immunity. For these diagnostic and therapeutic reasons, the demand for antibodies against Hu protein is rapidly growing. Methods: Polyclonal and monoclonal antibodies were generated using recombinant HuR protein. Western blot analyses were performed to check the specificity of generated antibodies using various recombinant proteins and cell lysates. Extracellular stimuli for HuR expression had been searched and HuR-associated proteins were isolated from polysome lysates and then separated in a 2-dimensional gel. Results: Polyclonal and monoclonal antibodies against HuR protein were generated and these antibodies showed HuR specificity. Antibodies were also useful to detect and immunoprecipitate endogenous HuR protein in Jurkat and BJAB. This report also revealed that TNF-${\alpha}$ treatment in BJAB up-regulated HuR expression. Lastly, protein profile in HuR-associated mRNAprotein complexes was mapped by 2-dimensional gel electrophoresis. Conclusion: This study reported that new antibodies against HuR protein were successfully generated. Currently, project to develop a diagnostic kit is in process. Also, this report showed that TNF-${\alpha}$ up-regulated HuR expression in BJAB and protein profile associated with HuR protein was mapped.

복제 소 태반과 IVF 소 태반의 protein pattern 분석

  • 김홍래;강재구;윤종택;성한우;조민래;박창식;진동일
    • Proceedings of the Korean Society of Developmental Biology Conference
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    • 2003.10a
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    • pp.90-90
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    • 2003
  • 체세포 핵이식에 의한 복제기술은 매우 낮은 성공률 나타내고 있어 실용화에 지장을 초래하고 있다. 이것은 후생적인 유전현상인 reprogramming이 불완전하게 이루어지기 때문인 것으로 추측되어지고 있다(Reik et al., Theriogenology 2003, 59: 21-32; Han et al, Theriogenology 2003, 59: 33-44). 체세포 핵이식 후에 태아사망의 원인이 태반의 비정상적인 기능과 관계가 있는 것으로 추정되는데 복제시 태아사망의 원인을 찾기 위해 본 연구를 시행하였다. 한우에서 체세포 복제 후 임신 말기에 태아가 사망한 태반조직 3개와 IVF 수정란 이식 후 동일한 시기에 제왕절개술을 실시한 태반조직 2개를 실험에 이용하였다. 태반 protein을 Two-Dimensional electrophoresis와 Mass spectrometer를 이용하여 분석 비교하였다. IPG-system을 이용하여 pH 4~7, pH 6~9에서 1차 전기영동을 한 후, 8~l6%의 SDS-PAGE gel에 2차 전기영동을 실시하였고 G-250 Coomassie로 염색하였다. gel 이미지는 Malanie III program을 이용하여 분석하였다. 전체 gel에서 약 1800개의 구분 가능한 protein spot이 나타났다. pH 4~7 범위에서 양적으로 차이나는 것 15개 중 복제한우 태반에서 증가되는 protein spot 5개와 감소하는 protein spot 10개를 골라 protein identification을 실시하였다. MALDI-TOF-MS를 이용하여 동정한 결과 phosphatidylinositol transfer protein-$\alpha$와 interleukin-18 등의 protein이 복제태반에서 발현이 증가되었고, 복제한우에서 발현이 감소되는 것으로는 vimentin, Rho-GDI-$\beta$, TRAST $\beta$-chain, ovarian sterol carrier protein 2, triosephosphate isomerase, tropemyosin beta chain, Aldose reductase 등으로 나타났다. 이러한 protein들은 inositol 지질 신호전달과 면역시스템, 세포분열, 산소 운반, steroidogenic 세포에서의 콜레스테롤 이동, 촉매 작용, 대사 작용 등에 중요한 역할을 하는 것으로 알려져 체세포 복제에 의한 태아사망 원인은 태반에서 이러한 protein들의 비정상적인 발현에 기인된 것으로 추정된다.

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Antioxidant Effects of Tocotrienol in Rice Bran (미강 함유 Tocotrienol의 항산화 효과)

  • Woo Ki-Min;Lee Young-Sang;Kim Yong-Ho
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.50 no.spc1
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    • pp.4-7
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    • 2005
  • The pharmaceutical function of tocotrienol in rice bran was evaluated. Distinctive antioxidative effects by 1,1-diphenyl-2-picrylhydrazyl(DPPH) could be observed. Also, Superoxide Dismutase(SOD) and Glutathione Peroxidase(GPX) activities of the cultured cells such as human firbroblast and hepatocyte, were increased up to 2 fold by the treatment of tocotrienol. The effects on GPX activity were more evident than SOD activity, and the stimulation was up to 2 fold. The changes of gene expression patterns were examined by applying the cell extracts of fibroblast treated with the increasing concentrations of tocotrienol on two-dimensional gel electrophoresis(2-D gel electrophoresis). As the concentrations increasing, many proteins began to appear with the increasing amounts, while several proteins diminished or disappeared. From these results, tocotrienol was clearly shown to have abilities on protecting any oxidizing damages and stimulating anti-oxidizing activities of the organisms.

Identification of a Marker Protein for Cardiac Ischemia and Reperfusion Injury by Two-Dimensional Gel Electrophoresis and Matrix-Assisted Laser Desorption Ionization Mass Spectrometry

  • Lee, Young-Suk;Kim, Na-Ri;Kim, Hyun-Ju;Joo, Hyun;Kim, Young-Nam;Jeong, Dae-Hoon;Cuong, Dang Van;Kim, Eui-Yong;Hur, Dae-Young;Park, Young-Shik;Hong, Yong-Geun;Lee, Sang-Kyung;Chung, Joon-Yong;Seog, Dae-Hyun;Han, Jin
    • The Korean Journal of Physiology and Pharmacology
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    • v.8 no.4
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    • pp.207-211
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    • 2004
  • The purpose of the present study was to evaluate the expression of cardiac marker protein in rabbit cardiac tissue that was exposed to ischemic preconditioning (IPC), or ischemiareperfusion injury (IR) using two-dimensional gel electrophoresis (2DE) and matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS). We compared 2DE gels of control (uninjured) cardiac tissue with those of IPC and IR cardiac tissue. Expression of one protein was detected in IR heart tissue, however the protein was not detected in the samples of control and IPC tissue. To further characterize the detected protein molecule, the protein in the 2D gel was isolated and subjected to trypsin digestion, followed by MALDI-MS. The protein was identified as myoglobin, which was confirmed also by Western blot analysis. These results are consistent with previous studies of cardiac markers in ischemic hearts, indicating myoglobin as a suitable marker of myocardial injury. In addition, the present use of multiple techniques indicates that proteomic analysis is an appropriate means to identify cardiac markers in studies of IPC and IR.

Proteome Approach as a Tool for the Efficient Separation of Seed Storage Proteins from Buckwheat

  • Cho, Seong-Woo;Kwon, Soo-Jeong;Roy, Swapan Kumar;Woo, Sun-Hee
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.60 no.1
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    • pp.29-32
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    • 2015
  • Two-dimensional electrophoresis (2-DE) was executed to separate the seed storage proteins from the buckwheat. The proteins extracted from the whole seed proteins were better separated and observed in the use of lysis buffer. Using this method, the highly reproducible isoelectric focusing (IEF) can be obtained from polyacrylamide gels, and IEF from the polyacrylamide gel at all the possible pH range (5.0-8.0) was more easily separated than IPG (immobilized pH gradient) gels. The polyacrylamide gels in the first dimension in 2-DE was used to separate and identify a number of whole seed proteins in the proteome analysis. In this new apparatus using 2-DE, 27cm in length of plate coated with polyacrylamide gel was used and the experiment was further investigated under the various conditions.

Analysis of outer mombrane proteins of Brucella abortus using two dimensional polyacrylamide gel electrophoresis (2차원 전기영동법을 이용한 Brucella abortus 세포외막 특이단백질의 분석)

  • Kim, Byung-su;Kim, Sun-hee;Kim, Jong-suk;Baek, Byeong-kirl
    • Korean Journal of Veterinary Research
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    • v.38 no.2
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    • pp.328-335
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    • 1998
  • Outer membrane proteins(OMPs) of Brucella abortus 1119-3 strain were extracted by Triton X-100 treatment, and fractionated by DEAE-cellulose column chromatography and Sephacryl S-300 column chromatography. The antigenic proteins in these fractions were identified by Western blot analysis. In Western blot analysis, a single band(38kDa) was observed in the DEAE fractions from 36th fraction to 38th fraction against sera of cattle infected with B abortus. And other fractions have several bands. However, the Sephacryl S-300 fractions exhibited a total of 3 peaks of proteins with a broad range from about 30 to 116kDa. In order to characterize further, the extracted OMPs and the DEAE fractions were analyzed by two dimensional polyacrylamide gel electrophoresis(2-DE) and Western blot using serum from naturally infected cattle with Brucella spp. The 2-DE immunoblots of DEAE fraction showed immunoreactive spots more than twenty two. The major protein spots have ranging from about 32 to 47kDa. The pI values of the spots were detected from pH 4.7 to 5.4. Among the major protein spots, the 38kDa protein which is a specific antigen, located at the point of approximately a pI 4.8.

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Microcontact Printing of Bacteria Using Hybrid Agarose Gel Stamp (혼성 아가로즈젤 스탬프를 이용한 박테리아 마이크로 컨택트 프린팅)

  • Shim, Hyun-Woo;Lee, Ji-Hye;Lee, Chang-Soo
    • KSBB Journal
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    • v.21 no.4
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    • pp.273-278
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    • 2006
  • The noble method of hybrid agarose gel microstamp fabricated by replica molding against PDMS master to make bacteria patterns on agar surface was presented. After the fabricated hybrid agarose gel microstamp was inked with E. coli, we could obtain 2 dimensional bacterial arrays with $50{\mu}m$ circular spots. And the various shaped patterns based on experimental design were easily generated. The analysis of mean fluorescent signal was showed that bacterial pattern have high contrast between spots and background and homogeneity of pattern. Our proposed method solved the problem of wetting and handling with small soft agarose gel microstamp when bacteria were used for ink. The agarose gel stamp provides appropriate environment to inked bacteria, which is essential technology for cell patterning with high retaining viability during the patterning process. This method is reproducible, convenient, rapid, and could be applied to screening system, study of cell-surface interaction, and microbial ecology.