• Title/Summary/Keyword: cell library

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In Vitro Selection of Cancer-Specific RNA Aptamers

  • Lee Young-Ju;Lee Seong-Wook
    • Journal of Microbiology and Biotechnology
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    • v.16 no.7
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    • pp.1149-1153
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    • 2006
  • In this study, nuclease-resistant RNA aptamers that are specific for Jurkat T leukemia cells were selected by a subtractive systemic evolution of ligands by exponential enrichment (SELEX) method. A randomized nuclease-resistant RNA library was incubated with normal peripheral blood mononuclear cells (PBMC) in each round to preclude RNAs that recognize the common cellular components on the surface of normal and cancer cells. The precluded RNAs were used for the selection of Jurkat T cell-specific aptamers, and the specific RNAs were then gradually enriched from start to the following selections. After 16 rounds of the subtractive SELEX, the selected aptamers were found to preferentially bind to Jurkat T cells, but not to the normal PBMC, evidenced by fluorescence-activated cell sorting analysis. Thus, the subtractive SELEX can be used to identify ligands to cancer-specific biological markers without prior knowledge of the nature of markers. The aptamers could be applied to specific cell sorting, tumor therapy, and diagnosis, and moreover, to find cancer cell-specific markers.

A Circuit Model of PEMFC for Design and Analyze Fuel Cell Power System (연료전지 전원 시스템의 설계 및 분석을 위한 PEMFC의 회로 모델)

  • Lee S.H.;Lee H.W.;Kwon S.K.
    • Proceedings of the KIPE Conference
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    • 2006.06a
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    • pp.197-199
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    • 2006
  • The Proton Exchange Membrane Fuel Cells (PEMFCs) are being used in a variety of applications including portable power generation, transportation and back-up power systems. In this paper presents a novel circuit model for a PEMFC that can be used to design and analyze fuel-cell power system. The Pspice-based model uses BJTs, L and C elements available in the Pspice library with some modification. The model includes the phenomena like activation polarization, ohmic polarization and mass transport effect present in a PEM fuel cell. Simulated characteristics of the fuel cell were compared with the experimental results obtained on a commercial fuel cell.

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Mapping Cellular Coordinates through Advances in Spatial Transcriptomics Technology

  • Teves, Joji Marie;Won, Kyoung Jae
    • Molecules and Cells
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    • v.43 no.7
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    • pp.591-599
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    • 2020
  • Complex cell-to-cell communication underlies the basic processes essential for homeostasis in the given tissue architecture. Obtaining quantitative gene-expression of cells in their native context has significantly advanced through single-cell RNA sequencing technologies along with mechanical and enzymatic tissue manipulation. This approach, however, is largely reliant on the physical dissociation of individual cells from the tissue, thus, resulting in a library with unaccounted positional information. To overcome this, positional information can be obtained by integrating imaging and positional barcoding. Collectively, spatial transcriptomics strategies provide tissue architecture-dependent as well as position-dependent cellular functions. This review discusses the current technologies for spatial transcriptomics ranging from the methods combining mechanical dissociation and single-cell RNA sequencing to computational spatial re-mapping.

HBV-X 단백질과 간세포 단백질간의 상호작용

  • 윤영대;김민재;김학대;이현숙;허윤실
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1993.04a
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    • pp.64-64
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    • 1993
  • 당해년도에는 HBV-X와 결합하는 상대 단백질을 탐색하는 것이 주목적이다. 이 목적으로 1) 우선 생물학적 활성을 보유하는 X-단백질을 E. coil system에서 다량으로 생산하는 공정을 확립하였으며 2) 이 X-단백질을 labelling한 후 Probe로 사용하여 liver cell내에 존재하는 43Kd, 48Kd, 55Kd, 100Kd의 단백질이 HBV-X에 결합하는 것을 확인하였으며 3) liver cell expression library를 screening하여 HBV-X와 결합하는 단백질을 coding하는 유전자를 cloning하여 현재 각 clone들을 규명하고 있는 중이다. 4) 또한 암억제 유전자 산물인 p53과 X-단백질과의 상호작용을 발견하였다. 이러한 결과는 X-단백질이 간암의 발생에 작용하는 기작을 설명할 수 있는 중요한 발견이다.

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Nervonic Acid Inhibits Replicative Senescence of Human Wharton's Jelly-Derived Mesenchymal Stem Cells

  • Sun Jeong Kim;Soojin Kwon;Soobeen Chung;Eun Joo Lee;Sang Eon Park;Suk-Joo Choi;Soo-Young Oh;Gyu Ha Ryu;Hong Bae Jeon;Jong Wook Chang
    • International Journal of Stem Cells
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    • v.17 no.1
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    • pp.80-90
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    • 2024
  • Cellular senescence causes cell cycle arrest and promotes permanent cessation of proliferation. Since the senescence of mesenchymal stem cells (MSCs) reduces proliferation and multipotency and increases immunogenicity, aged MSCs are not suitable for cell therapy. Therefore, it is important to inhibit cellular senescence in MSCs. It has recently been reported that metabolites can control aging diseases. Therefore, we aimed to identify novel metabolites that regulate the replicative senescence in MSCs. Using a fecal metabolites library, we identified nervonic acid (NA) as a candidate metabolite for replicative senescence regulation. In replicative senescent MSCs, NA reduced senescence-associated 𝛽-galactosidase positive cells, the expression of senescence-related genes, as well as increased stemness and adipogenesis. Moreover, in non-senescent MSCs, NA treatment delayed senescence caused by sequential subculture and promoted proliferation. We confirmed, for the first time, that NA delayed and inhibited cellular senescence. Considering optimal concentration, duration, and timing of drug treatment, NA is a novel potential metabolite that can be used in the development of technologies that regulate cellular senescence.

Anti-proliferative Effects and Apoptosis Induced by Chrysin or Emodin in Human Colorectal HCT116 Cells (Chrysin과 emodin에 의한 대장암 세포 항 성장 활성 및 세포사멸)

  • Ryu, Seung-Min;Kim, Yong-Hyun;Lee, Eun-Joo;Chung, Chungwook;Kim, Jong-Sik
    • Journal of Life Science
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    • v.31 no.10
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    • pp.929-936
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    • 2021
  • In the present study, we screened candidate natural compounds which possess the strong anti-proliferative effects on human colorectal HCT116 cells using the commercial natural product library (Selleckchem, L1400) based on cell viability assay. Human colorectal cancer HCT116 cells were incubated with 50 μM of each compound from the natural product library, and then cell viability was measured by MTT assay. From the first screening, five different kinds of natural products (chrysin, diosmetin, emodin, piperlongumine, and tanshinone I) were selected based on cell viability assay in HCT116 cells and commercial availability. All selected natural products significantly decreased cell viabilities in HCT116 cells, whereas pro-apoptotic protein NAG-1 is strongly induced by chrysin or emodin treatment. Chrysin and emodin decreased cell viability in a dose-dependent manner. Moreover, chrysin and emodin increased the expression of pro-apoptotic NAG-1 protein in a dose- and time-dependent manner. In addition, PARP cleavage induced by chrysin or emodin was recovered in part by the transfection of NAG-1 siRNA indicating that NAG-1 may be one of the genes responsible for apoptosis induced by chrysin or emodin. Overall, our findings may provide basic screening data on natural products which possess anti-proliferative activities and may help to understand the molecular mechanisms of anti-proliferative and pro-apoptotic activities mediated by chrysin and emodin.

Cyclosporine A and bromocriptine attenuate cell death mediated by intracellular calcium mobilization

  • Kim, In-Ki;Park, So-Jung;Park, Jhang-Ho;Lee, Seung-Ho;Hong, Sung-Eun;Reed, John C.
    • BMB Reports
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    • v.45 no.8
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    • pp.482-487
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    • 2012
  • To identify the novel inhibitors of endoplasmic reticulum stress-induced cell death, we performed a high throughput assay with a chemical library containing a total of 3,280 bioactive small molecules. Cyclosporine A and bromocriptine were identified as potent inhibitors of thapsigargiin-induced cell death (cut-off at $4{\sigma}$ standard score). However, U74389G, the potent inhibitor of lipid peroxidation had lower activity in inhibiting cell death. The inhibition effect of cyclosporine A and bromocriptine was specific for only thapsigargin-induced cell death. The mechanism of inhibition by these compounds was identified as modification of the expression of glucose regulated protein-78 (GRP-78/Bip) and inhibition of phosphorylation of p38 mitogen activated protein kinase (MAPK). However, these compounds did not inhibit the same events triggered by tunicamycin, which was in agreement with the cell survival data. We suggest that the induction of protective unfolded protein response by these compounds confers resistance to cell death. In summary, we identified compounds that may provide insights on cell death mechanisms stimulated by ER stress.

VLSI Design of High Speed Digital Neural Network using the Binary Convolution (Binar Convolution을 이용한 고속 디지탈 신경회로망의 VLSI 설계)

  • Choi, Seung-Ho;Kim, Young-Min
    • The Journal of the Acoustical Society of Korea
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    • v.15 no.5
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    • pp.13-20
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    • 1996
  • Recently, for implementation of neural networks extensive studies have been done especially VLSI technology has been regarded as the one of the most attractive means to implement neural networks. The main drawbacks of digital VLSI implementations are their large area and slow processing speed. In this paper to solve the speed and size problems we designed the efficient architecture using the binary convolution method for basic operation of neural cell, multiplication and addition. When it is used for implementing 3-layer network with 16 neural cell per layer that used neural cell based on binary convolution, clock of 50MHz and 26MCPS on 0.8${\mu}$ standard cell library has been achieved.

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Silencing of CaCDPK4 ( Capsicum annuum Calcium Dependent Protein Kinase) and ItsOrtholog, NbCDPK5 Induces Cell Death in Nicotiana benthamiana

  • Eunsook Chung;Kim, Young-Cheol;Oh, Sang-Keun;Younghee Jung;Kim, Soo-Yong;Park, Doil
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.77.1-77
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    • 2003
  • We have isolated a full-length cDNA clone, CaCDPK4 encoding a typical calcium-dependent protein kinase (CDPK) from hot pepper cDNA library. Genomic southern blot analysis showed that it belongs to a multigene family, but represents a single copy gone in hot pepper genome. RNA expression pattern of this gene revealed that it is induced by infiltration of Xanthomonas axonopodis pv. glycines Bra into hot pepper leaves but not by water deficit stress. However, high salt treatment of NaCl (0.4 M) solution to hot pepper plants strongly induced CaCDPK4 gene. In addition, this gene is weakly responsive to the exogenous application of salicylic acid or ethephon. Biochemical study of the GST-CaCDPK4 recominant protein showed that it autophosphorylates in vitro and the presence of EGTA, a calcium chelater, eliminates the kinase activity of the recombinant protein. As a way to identify the in vivo function of CaCDPK4 in plants, VIGS (Virus-Induced Gene Silencing) was employed. Agrobacterium-mediated TRV silencing construct containing the kinase and calmodulin domain of CaCDPK4 resulted in cell death of Nicotiana benthamiana plants. A highly homologous H benthamiana CDPK gene, NbCDPK5, to CaCDPK4 was cloned from N. benthamiana cDNA library. VIGS of NbCDPK5 also resulted in cell death. The molecular characterization of this cell death phenotype is being under investigation.

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Development of a Novel Vector System for Programmed Cell Lysis in Escherichia coli

  • Yun, Ji-Ae;Park, Ji-Hye;Park, Nan-Joo;Kang, Seo-Won;Ryu, Sang-Ryeol
    • Journal of Microbiology and Biotechnology
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    • v.17 no.7
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    • pp.1162-1168
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    • 2007
  • Although widely used as a host for recombinant protein production, Escherichia coli is unsuitable for massive screening of recombinant clones, owing to its poor secretion of proteins. A vector system containing T4 holin and T7 lysozyme genes under the control of the ptsG promoter derivative that is inducible in the absence of glucose was developed for programmed cell lysis of E. coli. Because E. coli harboring the vector grows well in the presence of glucose, but is lysed upon glucose exhaustion, the activity of the foreign gene expressed in E. coli can be monitored easily without an additional step for cell disruption after the foreign gene is expressed sufficiently with an appropriate concentration of glucose. The effectiveness of the vector was demonstrated by efficient screening of the amylase gene from a Bacillus subtilis genomic library. This vector system is expected to provide a more efficient and economic screening of bioactive products from DNA libraries in large quantities.