• Title/Summary/Keyword: Cell Characterization

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Engineered microdevices for single cell immunological assay

  • Choi, Jong-Hoon
    • Interdisciplinary Bio Central
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    • v.2 no.2
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    • pp.1.1-1.8
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    • 2010
  • Microdevices have been used as effective experimental tools for the rapid and multiplexed analysis of individual cells in single-cell assays. Technological advances for miniaturizing such systems and the optimization of delicate controls in micron-sized space homing cells have motivated many researchers from diverse fields (e.g., cancer research, stem cell research, therapeutic agent development, etc.) to employ microtools in their scientific research. Microtools allow high-throughput, multiplexed analysis of single cells, and they are not limited by the lack of large samples. These characteristics may significantly benefit the study of immune cells, where the number of cells available for testing is usually limited. In this review, I present an overview of several microtools that are currently available for single-cell analyses in two popular formats: microarrays and microfluidic microdevices. Then, I discuss the potential to study human immunology on the single-cell level, and I highlight several recent examples of immunoassays performed with single-cell microdevice assays. Finally, I discuss the outlook for the development of optimized assay platforms to study human immune cells. The development and application of microdevices for studies on single immune cells presents novel opportunities for the qualitative and quantitative characterization of immune cells and may lead to a comprehensive understanding of fundamental aspects of human immunology.

Requirement of CD4 Help for Induction of CD8 T Cell Response Specific for Virally Derived H60

  • Ryu, Su-Jeong;Kang, Bo-Ra;Kim, Seok-Ho;Kim, Tae-Woo;Chang, Jun;Choi, Eun-Young
    • IMMUNE NETWORK
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    • v.12 no.3
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    • pp.118-125
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    • 2012
  • CD40-CD40L-mediated help from CD4 T cells is essential to induce primary CD8 T cell responses specific to the non-inflammatory cell-based antigen H60. In this study, using H60 as a model antigen, we generated recombinant vaccinia viruses (rVVs) expressing the H60 CD8 epitope and investigated whether CD4 help was required to activate the CD8 T cell response specific to the virally expressed H60. The immune response after infection with rVVs expressing H60 was similar to that after immunization with H60 congenic splenocytes, with a peak frequency of H60-specific CD8 T cells detected in the blood on day 10 post-infection. A CD8 T cell response specific for virally derived H60 was not induced in CD4-depleted mice, but was in CD40-deficient mice. These results provide insights into the characterization of the CD8 T cell response specifically for antigens originating from cellular sources compared to viral sources.

Cloning and Characterization of the Mycobacterium bovis BCG panB Gene Encoding Ketopantoate Hydroxymethyltransferase

  • Kim, Jin-Koo;Kim, Kwang-Dong;Lim, Jong-Seok;Lee, Hee-Gu;Kim, Sang-Jae;Cho, Sang-Hyun;Jeong, Won-Hwa;Choe, In-Seong;Chung, Thi-Wha;Paik, Sang-Gi;Choe, Yong-Kyung
    • BMB Reports
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    • v.34 no.4
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    • pp.342-346
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    • 2001
  • The Mycobacterium bovis BCG panB gene, encoding ketopantoate hydroxymethyltransferase (KPHMT), was cloned from a ${\lambda}gt11$ genomic library and sequenced. The DNA sequence encodes a protein that contains 281 amino acid residues (M, 29,337) with a high similarity to the KPHMTs. Subcloning of a 846 by open reading frame (ORF), but not a 735 by ORF, into the vector pUC19 led to complementation of the panB mutant of Escherichia coli. The BCG pang gene was overexpressed in E. coli and the KPHMT purified to homogeneity The recombinant protein was further confirmed by an enzymatic assay.

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Functional Characterization of ABCB4 Mutations Found in Low Phospholipid-Associated Cholelithiasis (LPAC)

  • Kim, Tae Hee;Park, Hyo Jin;Choi, Ji Ha
    • The Korean Journal of Physiology and Pharmacology
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    • v.17 no.6
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    • pp.525-530
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    • 2013
  • Multidrug resistance 3 (MDR3) is expressed on the canalicular membrane of the hepatocytes and plays an important role in protecting the liver from bile acids. Altered ABCB4 gene expression can lead to a rare hepatic disease, low phospholipid-associated cholelithiasis (LPAC). In this study, we characterized 3 ABCB4 mutations in LPAC patients using various in vitro assay systems. We first measured the ability of each mutant to transport paclitaxel and then the mechanisms by which these mutations might change MDR3 transport activity were determined using immunoblotting, cell surface protein biotinylation, and immunofluorescence. Through a membrane vesicular transport assay, we observed that the uptake of paclitaxel was significantly reduced in membrane vesicles expressing 2 ABCB4 mutations, F165I and S320F. Both mutants showed significantly decreased total and cell surface MDR3 expression. These data suggest two missense mutations of ABCB4 may alter function of MDR3 and ultimately can be determined as LPAC-causing mutations.

Fabrication and Characterization of Array Tactile Actuator Based on Cellulose Acetate (셀룰로오스 아세테이트 기반 어레이 촉각 액추에이터의 제작 및 특성평가)

  • Kim, Hyun-Chan;Yun, Sungryl;Ko, Hyun-U;Kim, Jaehwan
    • Journal of the Korean Society for Precision Engineering
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    • v.32 no.8
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    • pp.743-748
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    • 2015
  • This paper reports the enhanced fabrication and characterization of a $3{\times}3$ array tactile actuator composed of cellulose acetate. The array tactile actuator, with dimensions of $15{\times}15{\times}1mm^3$, consists of 9 pillar-supported cells made from a cellulose-acetate molding. The fabrication process and performance test along with the results for the suggested actuator are explained. To improve the cell-array fabrication, a laser cut was adopted after the molding process. The displacement of the unit cell increased the input voltage and frequency. Various top masses are added onto the actuator to mimic the touch force, and the acceleration of the actuator is measured under actuation. When 2 kV is applied to the actuator, the maximum acceleration is 0.64 g, which is above the vibrotactile threshold. The actuation mechanism is associated with the electrostatic force between the top and bottom electrodes.

Characterization of Nafion/Pt/Polypyrrole Composite Membrane Prepared by Chemical In-situ Polymerization for DMFC (화학적 합성에 의해 제조된 직접 메탄올 연료전지용 나피온/백금/폴리피롤 복합 막의 특성 분석)

  • Park, Ho-Seok;Kim, Yo-Jin;Im, Hun-Suk;Choi, Bong-Gill;Hong, Won-Hi
    • 한국신재생에너지학회:학술대회논문집
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    • 2006.11a
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    • pp.421-424
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    • 2006
  • Nafion/Pt/Polypyrrole composite membranes were fabricated by chemical in-situ polymerization of pyrrole monomers with Pt precursors in Nafion matrix for DMFC. We demonstrated that positively charged pyrrolinum groups of polypyrrole particles were co-interacted with sulfonic groups of Nafion as verified by FT-IR results. Mutual interaction between $Nafion-SO_3^-$ (or negatively charged Pt precursors) and Polypyrrole$-NH_2^+$ influenced the physical properties of pristine Nafion. Thermal property proton conductivity, methanol permeability, and cell performance of pristine and modified Nafion were analyzed for an application of DMFC membrane. Thermal stabilities of sulfonic groups and side chains in Nafion/Pt/polypyrrole composite membranes were higher than those of Nafion due to mutual interaction between sulfonic groups of Nafion and pyrrolinum groups of polypyrrole. Methanol permeabilities of Nafion/Pt/Polypyrrole composite were reduced more proton conductivities with the increase in the content of Pt particles. As a result of that, the enhancement of cell performance by Nafion/Pt/Polypyrole O2 relative to Nafion was more pronounced under the specific experimental condition such as high temperature and more concentrated methanol solution.

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Purification and Biochemical Characterization of Recombinant Alanine Dehydrogenase fvom Thermus caldophilux GK24

  • Bae, Jung-Don;Cho, Youn-Jeung;Kim, Doo-Il;Lee, Dae-Sil;Shin, Hyun-Jae
    • Journal of Microbiology and Biotechnology
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    • v.13 no.4
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    • pp.628-631
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    • 2003
  • The recombinant alanine dehydrogenase (ADH) from E. coli containing Thermus caldophilus ADH was purified to homogeneity from a cell-free extract. The enzyme was purified 38-fold with a yield of 68% from the starting cell-free extract. The purified enzyme gave a single band in polyacrylamide gel electrophoresis, and its molecular weight was estimated to be 45 kDa. The pH optimum was 8.0 for reductive amination of pyruvate and 12.0 for oxidative deamination of L-alanine. The enzyme was stable up to $70^{\circ}C$. The activity of the enzyme was inhibited by 1 mM $Zn^{2+}$, 20% hexane, and 20% $CHCl_3$. However, 10 mM $Mg^{2+}$ and 40% propanol had no effect on the enzyme activity. The Michaelis constants ($K_m$) for the substrates were $50\;\mu\textrm{M}$ for NADH, 0.2 mM for pyruvate, 39.4 mM for $NH_4+$, 2.6 mM for L-alanine, and 1.8 mM for $NAD^+$.

Effect of Carbon Support (CNTs) on Pt/Au/TiO2 Catalyst Preparation and Characterization for Direct Methanol Fuel Cell (DMFC) (탄소 담지체(CNTs)에 따른 직접메탄올연료전지용 Pt/Au/TiO2 촉매 제조와 평가)

  • You, Sun-Kyung;Kim, Han-Joo;Kim, Tae-Il;Kang, Kyung-Suk;Park, Soo-Gil
    • Journal of the Korean Electrochemical Society
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    • v.12 no.3
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    • pp.282-286
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    • 2009
  • The synthesis and characterization of catalysts supported on multi-walled carbon nanotubes (CNTs). $Pt/Au/TiO_2$ is added to a CNTs(cabon nano tube) carbon support to improve the performance of a direct methanol fuel cell. XRD and SEM showed that uniform anatase $TiO_2$ and Pt/Au particles were about 200 nm and 20${\sim}$25 nm in diameter. The composite catalyst activities were measured by cyclic voltammetry (CV), demonstrating that it is more promising for use in fuel cells.

Message in a Bottle: Chemical Biology of Induced Disease Resistance in Plants

  • Schreiber, Karl;Desveaux, Darrell
    • The Plant Pathology Journal
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    • v.24 no.3
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    • pp.245-268
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    • 2008
  • The outcome of plant-pathogen interactions is influenced significantly by endogenous small molecules that coordinate plant defence responses. There is currently tremendous scientific and commercial interest in identifying chemicals whose exogenous application activates plant defences and affords protection from pathogen infection. In this review, we provide a survey of compounds known to induce disease resistance in plants, with particular emphasis on how each compound was originally identified, its putative or demonstrated mechanism of defence induction, and the known biological target(s) of each chemical. Larger polymeric structures and peptides/proteins are also discussed in this context. The quest for novel defence-inducing molecules would be aided by the capability for high-throughput analysis of candidate compounds, and we describe some issues associated with the development of these types of screens. Subsequent characterization of hits can be a formidable challenge, especially in terms of identifying chemical targets in plant cells. A variety of powerful molecular tools are available for this characterization, not only to provide insight into methods of plant defence activation, but also to probe fundamental biological processes. Furthermore, these investigations can reveal molecules with significant commercial potential as crop protectants, although a number of factors must be considered for this potential to be realized. By highlighting recent progress in the application of chemical biology techniques for the modulation of plant-pathogen interactions, we provide some perspective on the exciting opportunities for future progress in this field of research.

Electrical Characterization of Cu(InxGa1-x)(SySe2-y) Thin Film Solar Cells

  • Kim, Dahye;Kim, Ji Eun;Cho, Yunae;Kim, Dong-Wook
    • Proceedings of the Korean Vacuum Society Conference
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    • 2014.02a
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    • pp.464.1-464.1
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    • 2014
  • Among numerous material candidates, Cu(InxGa1-x)(SySe2-y) (CIGS) thin films have emerged as promising material candidates for thin film solar cell applications due to the high energy conversion efficiency and relatively low fabrication cost. The CIGS thin film solar cells consist of several materials, including Mo back contacts, ZnO-based window layers, and CdS buffer layers. All these materials have different crystal structures and contain quite distinct chemical elements, and hence the device characterization requires careful analyses. Most of all, identification of the major trap states resulting in the carrier recombination processes is a key step toward realization of high efficiency CIGS solar cells. We have carried out electrical investigations of CIGS thin film solar cells to specify the major trap states and their roles in photovoltaic performance. In particular, we have used the temperature-dependent transport characterizations and admittance spectroscopy. In this presentation, we will introduce some exemplary studies of DC and AC electrical characteristics of the CIGS solar cells.

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