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Single Cell Array of Biotinylated Cells Using Surface Functionalization and Microcontact Printing
Lee, Zee-Won,Lee, Kyung-Bok,Hong, Jang-Hee,Kim, Jae-Hong,Choi, Inpyo,Choi, Insung S. Chemical Society of Japan 2005 Chemistry letters Vol.34 No.5
<P>This paper describes a versatile method for generating single cell arrays on a glass substrate, which could be applicable to any arbitrary cell types, by a combination of surface functionalization, biotinylation of cells, and microcontact printing (μCP).</P>
Lee, Hye-Mi,Shin, Dong-Min,Choi, Dae-Kyoung,Lee, Zee-Won,Kim, Ki-Hye,Yuk, Jae-Min,Kim, Chang Deok,Lee, Jeung-Hoon,Jo, Eun-Kyeong Blackwell Publishing Ltd 2009 Cellular microbiology Vol.11 No.4
<P>Summary</P><P><I>Mycobacterium ulcerans</I> (MU), an environmental pathogen, causes Buruli ulcer, a severe skin disease. We hypothesized that epidermal keratinocytes might not be a simple barrier, but play a role during MU infection through pattern-recognition receptors expressed in keratinocytes. We found that keratinocyte Toll-like receptors (TLRs) 2 and 4 and Dectin-1 actively participate in the innate immune response to MU, which includes the internalization of bacteria, the production of reactive oxygen species (ROS), and the expression of chemokines and LL-37. Human keratinocytes constitutively expressed TLRs 2 and 4 and induced Dectin-1 in response to MU. Exposing keratinocytes to MU resulted in rapid ROS production, which in turn contributed to the mRNA and protein expression of LL-37. In addition, TLR2, Dectin-1 and, to an extent, TLR4 are essential for the MU-mediated expression of CXCL8, CCL2 and LL-37 in keratinocytes. Furthermore, confocal analysis showed that the Dectin-1 is necessary for keratinocytes to internalize bacilli. Importantly, blockade of ROS and LL-37 significantly increased the intracellular MU growth in keratinocytes, suggesting an important role of these mediators for cutaneous innate immune responses. Our results demonstrate that TLR2, TLR4 and Dectin-1 actively sense, internalize and respond in an innate way to MU in human epidermal keratinocytes.</P>
Lee, Sang Kwang,Kim, Yongtae,Kim, Sung-Soo,Lee, Jeong Hwa,Cho, Kun,Lee, Sang Sook,Lee, Zee-Won,Kwon, Kyung-Hoon,Kim, Young Hye,Suh-Kim, Haeyoung,Yoo, Jong Shin,Park, Young Mok WILEY-VCH Verlag 2009 Proteomics Vol.9 No.18
<P>Mesenchymal stem cells (MSCs) are multipotent cells, which have the capability to differentiate into various mesenchymal tissues such as bone, cartilage, fat, tendon, muscle, and marrow stroma. However, they lose the capability of multi-lineage differentiation after several passages. It is known that basic fibroblast growth factor (bFGF) increases growth rate, differentiation potential, and morphological changes of MSCs in vitro. In this report, we have used 2-DE coupled to MS to identify differentially expressed proteins at the cell membrane level in MSCs growing in bFGF containing medium. The cell surface proteins isolated by the biotin–avidin affinity column were separated by 2-DE in triplicate experiments. A total of 15 differentially expressed proteins were identified by quadrupole-time of flight tandem MS. Nine of the proteins were upregulated and six proteins were downregulated in the MSCs cultured with bFGF containing medium. The expression level of three actin-related proteins, F-actin-capping protein subunit alpha-1, actin-related protein 2/3 complex subunit 2, and myosin regulatory light chain 2, was confirmed by Western blot analysis. The results indicate that the expression levels of F-actin-capping protein subunit alpha-1, actin-related protein 2/3 complex subunit 2, and myosin regulatory light chain 2 are important in bFGF-induced morphological change of MSCs.</P>
Lee, Sang Kwang,Kim, Jae Ho,Kim, Sung-Soo,Kang, Taewook,Park, Nam Hyun,Kwon, Kyung-Hoon,Lee, Sang Sook,Lee, Zee Won,Suh-Kim, Hae young,Cho, Kun,Yun, Su Yeoung,Han, Ji Young,Yoo, Jong Shin,An, Hyun Joo Springer-Verlag 2013 Analytical and bioanalytical chemistry Vol.405 No.16
<P>Mulitpotent mesenchymal stem cells (MSCs) derived from human bone marrow are promising candidates for the development of cell therapeutic strategies. MSC surface protein profiles provide novel biological knowledge concerning the proliferation and differentiation of these cells, including the potential for identifying therapeutic targets. Basic fibroblast growth factor (bFGF) affects cell surface proteins, which are associated with increased growth rate, differentiation potential, as well as morphological changes of MSCs in vitro. Cell surface proteins were isolated using a biotinylation-mediated method and identified using a combination of one-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis and mass spectrometry. The resulting gel lines were cut into 20 bands and digested with trypsin. Each tryptic fragment was analyzed by liquid chromatography-electrospray ionization tandem mass spectrometry. Proteins were identified using the Mascot search program and the International Protein Index human database. Noble MSC surface proteins (n = 1,001) were identified from cells cultured either with (n = 857) or without (n = 667) bFGF-containing medium in three independent experiments. The proteins were classified using FatiGO to elucidate their function. We also confirmed the proteomics results using Western blotting and immunofluorescence microscopic analysis. The nature of the proteins identified makes it clear that MSCs express a wide variety of signaling molecules, including those related to cell differentiation. Among the latter proteins, four Ras-related Rab proteins, laminin-R, and three 14-3-3 proteins that were fractionated from MSCs cultured on bFGF-containing medium are implicated in bFGF-induced signal transduction of MSCs. Consequently, these finding provide insight into the understanding of the surface proteome of human MSCs.</P>
Effect of Botulinum Toxin Type A on Morphology of Salivary Glands in Patients with Cerebral Palsy
Zee-Ihn Lee,Dong-Hyun Cho,Won-Duck Choi,박동휘,Seung-Deuk Byun 대한재활의학회 2011 Annals of Rehabilitation Medicine Vol.35 No.5
Objective To investigate the eff ect of botulinum toxin type A (BTXA) on drooling and the morphologic change of the salivary gland in patients with cerebral palsy. Method Eight cerebral palsy patients suffering from severe drooling participated in this study. BTXA was injected into both submandibular and parotid glands under intravenous sedation and with ultrasound guidance (1 unit/gland/kg: maximum 100 units) in an outpatient or inpatient procedure. The severity of drooling was measured before injection and 3 weeks after injection using the Teacher Drooling Scale, the Drooling Score-severity,frequency and the Visual Analog Scale. To investigate the morphologic change of the salivary glands, the size of salivary glands were measured before injection and 3 weeks after injection using computed tomography of the neck. The measurement values were analyzed by Wilcoxon signed rank test. Results Statistically significant improvements were shown in all three parameters for assessing the severity of drooling after BTXA injections (p<0.05). Size of the salivary glands were signifi cantly decreased at 3 weeks after BTXA injection (p<0.05). Conclusion Salivary gland injection with BTXA could be a useful treatment method to reduce drooling in patients with cerebral palsy and decreased size of salivary glands may partially explain the mechanism.
Locus-Specific Reversible DNA Methylation Regulates Transient IL-10 Expression in Th1 Cells
Hwang, Won,Lee, Choong-Gu,Lee, Changhon,Verma, Ravi,Rudra, Dipayan,Park, Zee Yong,Im, Sin-Hyeog American Association of Immunologists 2018 Journal of Immunology Vol. No.
<P>IL-10 is a pleiotropic cytokine with multifaceted functions in establishing immune homeostasis. Although expressed by Th1 and Th2 cells, conventional Th1 cells produce marginal levels of IL-10 compared with their Th2 counterparts. In this study, we investigated the epigenetic mechanisms of <I>Il-10</I> gene expression in Th1 cells. Bioinformatics EMBOSS CpG plot analysis and bisulfite pyrosequencing revealed three CpG DNA methylation sites in the <I>Il-10</I> gene locus. Progressive DNA methylation at all of the CpG regions of interest (ROIs) established a repressive program of <I>Il-10</I> gene expression in Th1 cells. Interestingly, Th1 cells treated with IL-12 and IL-27 cytokines, thereby mimicking a chronic inflammatory condition in vivo, displayed a significant increase in IL-10 production that was accompanied by selective DNA demethylation at ROI 3 located in intron 3. IL-10–producing T cells isolated from lymphocytic choriomeningitis virus–infected mice also showed enhanced DNA demethylation at ROI 3. Binding of STAT1 and STAT3 to demethylated ROI 3 enhanced IL-10 expression in an IL-12/IL-27–dependent manner. Accordingly, CD4<SUP>+</SUP> T cells isolated from STAT1- or STAT3-knockout mice were significantly defective in IL-10 production. Our data suggest that, although stably maintained DNA methylation at the promoter may repress IL-10 expression in Th1 cells, locus-specific reversible DNA demethylation may serve as a threshold platform to control transient <I>Il-10</I> gene expression.</P>
Proteomic Analysis to Identify Tightly-Bound Cell Wall Protein in Rice Calli
Cho, Won Kyong,Hyun, Tae Kyung,Kumar, Dhinesh,Rim, Yeonggil,Chen, Xiong Yan,Jo, Yeonhwa,Kim, Suwha,Lee, Keun Woo,Park, Zee-Yong,Lucas, William J.,Kim, Jae-Yean Korean Society for Molecular and Cellular Biology 2015 Molecules and cells Vol.38 No.8
Rice is a model plant widely used for basic and applied research programs. Plant cell wall proteins play key roles in a broad range of biological processes. However, presently, knowledge on the rice cell wall proteome is rudimentary in nature. In the present study, the tightly-bound cell wall proteome of rice callus cultured cells using sequential extraction protocols was developed using mass spectrometry and bioinformatics methods, leading to the identification of 1568 candidate proteins. Based on bioinformatics analyses, 389 classical rice cell wall proteins, possessing a signal peptide, and 334 putative non-classical cell wall proteins, lacking a signal peptide, were identified. By combining previously established rice cell wall protein databases with current data for the classical rice cell wall proteins, a comprehensive rice cell wall proteome, comprised of 496 proteins, was constructed. A comparative analysis of the rice and Arabidopsis cell wall proteomes revealed a high level of homology, suggesting a predominant conservation between monocot and eudicot cell wall proteins. This study importantly increased information on cell wall proteins, which serves for future functional analyses of these identified rice cell wall proteins.
객체식별아이디 이포지션 기반의 LBSNS 앱이 19대 총선 후보 지지율의 변화에 미친 영향
이상지(Sang-Zee Lee),장동혁(Dong-Heyok Jang),박성운(Sung-Woon Park),조원희(Won-Hee Cho),이기철(Gi-Chul Yi) 한국콘텐츠학회 2013 한국콘텐츠학회논문지 Vol.13 No.8
19대 총선에서 특정 후보의 이름을 포함하도록 정한 통합식별아이디 이포지션(ePosition)을 기반으로 후보 개인 맞춤형 LBSNS 앱을 기획하여 개발하고 선거기간 동안 홍보 목적으로 활용하였다. 선거를 전후하여 본 LBSNS 앱이 후보의 지지율 변화에 어느 정도 기여했는지 정량적으로 그 영향을 분석하였다. 대전광역시 6개 지역구 24명의 후보를 대상으로 개인 맞춤형 LBSNS 앱을 활용한 후보와 그렇지 않은 후보를 구분하여 선거운동 기간 동안 언론에 공개적으로 발표된 지지율과 개표 결과를 바탕으로 개인별 지지율 변화를 비교하였다. 해당 앱을 활용한 3명의 후보는 각각 12.6%, 11.4% 및 11.2%씩 두 자리 수의 지지율 상승이 있었지만 나머지 21명의 후보들은 지지율 변화는 모두 3% 이내로 머물러 개인맞춤형 스마트폰 앱을 활용함으로써 후보 지지율 상승에 상당한 효과가 있었다. During 19th general election the customized LBSNS(Location Based SNS) application for some candidates of the National Assembly planned and developed based on the object identification, ePosition, comprising the candidates name have been applied for an election campaign. The approval rating change before and after 19th election campaign period for each candidate was quantitatively studied how it would be affected by the candidate custom LBSNS application. Only 3 out of 24 candidates in 6 local electorates in the Daejon Metropolitan City have adopted the customized LBSNS application and the rest 21 candidates have not, whose approval rating change before and after an election campaign has been analyzed comparatively candidate by candidate. The approval rating for 3 candidates adopting LBSNS application went up by 12.6%, 11.4%, 11.2% respectively, but those for the rest 21 candidates all changed within 3%.