• Title/Summary/Keyword: Two-cell

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Single-cell RNA sequencing identifies distinct transcriptomic signatures between PMA/ionomycin- and αCD3/αCD28-activated primary human T cells

  • Jung Ho Lee;Brian H Lee;Soyoung Jeong;Christine Suh-Yun Joh;Hyo Jeong Nam;Hyun Seung Choi;Henry Sserwadda;Ji Won Oh;Chung-Gyu Park;Seon-Pil Jin;Hyun Je Kim
    • Genomics & Informatics
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    • v.21 no.2
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    • pp.18.1-18.11
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    • 2023
  • Immunologists have activated T cells in vitro using various stimulation methods, including phorbol myristate acetate (PMA)/ionomycin and αCD3/αCD28 agonistic antibodies. PMA stimulates protein kinase C, activating nuclear factor-κB, and ionomycin increases intracellular calcium levels, resulting in activation of nuclear factor of activated T cell. In contrast, αCD3/αCD28 agonistic antibodies activate T cells through ZAP-70, which phosphorylates linker for activation of T cell and SH2-domain-containing leukocyte protein of 76 kD. However, despite the use of these two different in vitro T cell activation methods for decades, the differential effects of chemical-based and antibody-based activation of primary human T cells have not yet been comprehensively described. Using single-cell RNA sequencing (scRNA-seq) technologies to analyze gene expression unbiasedly at the single-cell level, we compared the transcriptomic profiles of the non-physiological and physiological activation methods on human peripheral blood mononuclear cell-derived T cells from four independent donors. Remarkable transcriptomic differences in the expression of cytokines and their respective receptors were identified. We also identified activated CD4 T cell subsets (CD55+) enriched specifically by PMA/ionomycin activation. We believe this activated human T cell transcriptome atlas derived from two different activation methods will enhance our understanding, highlight the optimal use of these two in vitro T cell activation assays, and be applied as a reference standard when analyzing activated specific disease-originated T cells through scRNA-seq.

Visualization of two-phae flow by using transparent Proton Exchange Membrane Fuel Cell (고분자 전해질 연료전지 가시화 장치를 이용한 이상유동 현상 관찰)

  • Lee, Dong-Ryul;Bae, Joong-Myeon
    • 한국신재생에너지학회:학술대회논문집
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    • 2009.06a
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    • pp.374-377
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    • 2009
  • The operating temperature of Proton Exchange Membrane Fuel Cell (PEMFC) usually has to be limited under $100^{\circ}C$ to maintain the proper ionic conductivity. Therefore, the only product from reaction, water, is in the liquid phase. Two-phase flow makes the flow phenomenon in the channel difficult to understand and predict. Water blocking in the PEMFC channel or the pore of Gas Diffusion Layer (GDL), called flooding, is known as the main effect of PEMFC degradation. To analyze two-phase flow, the PEMFC with transparent acrylic plate was used. Two-phase flow patterns were observed by varying the current density. When the PEMFC is mounted horizontally, water in the cathode is mainly transported on the interface between the channel and GDL.

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Two Newly Recorded Ciliates, Oxytricha longigranulosa and o. marina (Ciliophora: Spirotrichea: Sporadotrichida) from Korea

  • Kwon, Choon-Bong;Shin, Mann-Kyoon
    • Animal Systematics, Evolution and Diversity
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    • v.24 no.1
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    • pp.81-88
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    • 2008
  • Two oxytrichid ciliates collected from the mosses and estuarine littoral in Korea were identified as Oxytricha longigranulosa Berger and Foissner, 1989 and O. marina Kahl, 1932. These species are reported for the first time from Korea. The description was based on living and protargol impregnated specimens. Diagnostic characters for each species are as follows. Oxytricha longigranulosa: Cell in vivo $80-115{\times}30-50{\mu}m$, mostly $90{\times}40{\mu}m$. Length/width ratio about 2.4/1. Cortical granules about $1{\times}1.5{\mu}m$ in size, colorless, arranged in short and discontinued longitudinal rows. Four frontoventral cirri. Adoral zone of membrane lies (AZM) covering 30-50% of cell length with 25-27 adoral membranelles (AM). Buccal area flat, typical Oxytricha pattern. Five transverse cirri, 19-23 right marginal cirri, 19-24 left marginal cirri, three caudal cirri, five dorsal kineties. Two macronuclear nodules 2 in number and spherical in shape, two micronuclei in number. Oxytricha marina: Cell in vivo $100-150{\times}30-60{\mu}m$. Cytoplasm colorless without cortical granules. Four frontoventral cirri. AZM covering 50% of cell length with 28-44 AMs, Buccal area flat, typical Oxytricha pattern. Five transverse cirri, 23-38 right marginal cirri, 19-25 left marginal cirri, three caudal cirri, five dorsal kineties. Two macronuclear nodules and spherical in shape, 1-5 micronuclei, mostly two in number.

Effects of Pluronic F-68 on Cell Growth of Digitalis lanata in Aqueous Two-Phase Systems

  • LEE , SANG-YOON;KIM, DONG-IL
    • Journal of Microbiology and Biotechnology
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    • v.14 no.6
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    • pp.1129-1133
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    • 2004
  • The effects of Pluronic F-68, a non-ionic surfactant, on the growth and physical characteristics of Digitalis lanata suspension cultures were investigated in aqueous two-phase systems (ATPSs) composed of $4.5\%$ polyethylene glycol (PEG) 20,000 and $2.8\%$ crude dextran. In the range of 0.1-10.0 g $1^{-1}$, Pluronic F-68 enhanced the maximum cell density in a medium with ATPSs, even though Pluronic F-68 did not affect cell growth in a normal growth medium. In terms of physical properties of ATPSs with cell suspension cultures, 0.2 g $1^{-1}$ of Pluronic F-68 reduced viscosity by up to $40\%$, while 0.1 g $1^{-1}$ of Pluronic F-68 significantly enhanced the oxygen transfer rate. In addition, we successfully performed aqueous two-phase cultivation in a 5-1 stirred tank bioreactor with 0.5 g $1^{-1}$ of Pluronic F-68, and discovered that cell growth in ATPSs was similar to that in normal growth medium.

Effects of Hydroxychloroquine Co-administered with Chemotherapeutic Agents on Malignant Glioma Cell Lines : in vitro Study

  • Park, Yong-Sook;Choi, Jae-Young;Chang, Jong-Hee;Park, Yong-Gou;Chang, Jin-Woo
    • Journal of Korean Neurosurgical Society
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    • v.38 no.1
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    • pp.47-53
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    • 2005
  • Objective : Anti-malaria drugs may modulate tumor resistance to chemotherapeutic agents, but it has not been proven effective in the treatment of malignant gliomas. The aim of this study was to determine whether adequate pre-clinical data on co-administration of chemotherapeutic agents with anti-malaria drugs on malignant cell lines could be obtained that would warrant its further potential consideration for use in a clinical trial for malignant gliomas. Methods : Two malignant glioma cell lines [U87MG, T98G] were treated with chemotherapeutic agents alone or with anti-malaria drugs. Cells were incubated with drugs for 4 days. Following the 4-day incubation, drug sensitivity assays were performed using 3-[4,5-dimethyl-2-thiazol-2-yl] 2,5-diphenyltetrazolium bromide [MTT] assay following optimization of experimental conditions for each cell lines and cell viability was calculated. Results : In all of four chemotherapeutic agents[doxorubicin. vincrisitne, nimustine, and cisplatin], the cell viability was found to be markedly decreased when hydroxychloroquine was co-administered on both U87MG and T98G cell lines. The two way analysis of variance[ANOVA] yielded a statistically significant two-sided p-value of 0.0033[doxorubicin], 0.0005[vincrisitne], 0.0007[nimustine], and 0.0003[cisplatin] on U87MG cell lines and 0.0006[doxorubicin], 0.0421[vincrisitne], 0.0317[nimustine], and 0.0001[cisplatin] on T98G cell lines, respectively. However, treatment with chloroquine and primaquine did not induce a decrease in cell viability on both U87MG and T98G cell lines. Conclusion : Our data support further consideration of the use of hydroxychloroquine prior to systemic chemotherapy to maximize its tumoricidal effect for patients with malignant gliomas.

Similarities of Tobacco Mosaic Virus-Induced Hypersensitive Cell Death and Copper-Induced Abiotic Cell Death in Tobacco

  • Oh, Sang-Keun;Cheong, Jong-Joo;Ingyu Hwang;Park, Doil
    • The Plant Pathology Journal
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    • v.15 no.1
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    • pp.8-13
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    • 1999
  • Hypersensitive cell death of plants during incompatible plant-pathogen interactions is one of the efficient defense mechanisms of plants against pathogen infections. For better understanding of the molecular mechanisms involved in the plant hypersensitive response (HR), TMV-induced biotic plant cell death and CuSO4-induced abiotic plant cell death were compared in terms of expression patterns of ten different defense-related genes as molecular markers. The genes include five pathogenesis-related protein genes, two plant secondary metabolite-associated genes, two oxidative stress-related genes and one wound-inducible gene isolated from tobacco. Northern blot analyses revealed that a same set of defense-related genes was induced during both biotic and abiotic cell death but with different time and magnitude. The expression of defense-related genes in tobacco plants was temporarily coincided with the time of cell death. However, when suspension cell cultures was used to monitor the expression of defense-related genes, different patterns of the gene expression were detected. This result implies that three are common and, in addition, also different branches of signaling pathways leading to the induced expression of defense-related genes in tobacco during the pathogen- and heavy metal-induced cell death.

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Design of Cell Frame Structure of Unit Cell for Molten Carbonate Fuel Cell Using CFD Analysis (CFD를 통한 용융탄산염 연료전지 단위전지용 셀 프레임 구조 설계)

  • LEE, SUNG-JOO;LIM, CHI-YOUNG;LEE, CHANG-WHAN
    • Transactions of the Korean hydrogen and new energy society
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    • v.29 no.1
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    • pp.56-63
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    • 2018
  • In this study, a $100cm^2$ cell frame for a molten carbonate fuel cell was designed using CFD analysis. Electrochemical reactions, gas flow, and the heat transfer in $100cm^2$ cell frame were modeled using COMSOL Multiphysics. Two design variables such as the height of the cell frame and the length of the gas input area were determined to obtain minimized temperature distribution and uniform gas distribution. With two design parameter such as height of the cell frame and the length of the gas flow channel, the temperature difference in the cell fame was decreased to $5^{\circ}C$ and the gas uniformity in the flow channel were achieved.

Two Cases of Non-Small Cell Lung Cancer Treated with Intravenous Cultivated Wild Ginseng Pharmacopuncture (산삼약침 혈맥주입을 시행한 비소세포성폐암 환자 2례)

  • Bang, Sun-Hwi;Kwon, Ki-Rok;Yoo, Hwa-Seung
    • Journal of Pharmacopuncture
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    • v.11 no.2
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    • pp.13-19
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    • 2008
  • Objectives To investigate the therapeutic effects of intravenous cultivated wild ginseng(Panax ginseng C.A. Meyer) pharmacopuncture(CWGP) in treating patients with non-small cell lung cancer(NSCLC). Design Prospective case series. Setting This study was conducted at the East-West Cancer Center of Dunsan Oriental Hospital, Daejeon University. Patients Two non-small cell lung cancer patients. Intervention Two non-small cell lung cancer patients were injected CWGP(20mL/day) mixed with 0.9% normal saline(100mL) intravenously. Each patient received a total of 16 and 9 cycles, respectively. One cycle is composed of 14 days. Outcome Measures The effect of intravenous CWGP was measured by scanning with computed tomography(CT) after every 2 cycle and Positron emission tomography- computed tomography(PET/CT) after every 6 cycles. Response and progression was evaluated using the Response Evaluation Criteria in Solid Tumors(RECIST) Committee classification of complete response(CR), partial response(PR), progressive disease(PD) and stable disease(SD). Results They were treated with intravenous CWGP for 8 and 5 months respectively. time later, each tumor remains stable disease(SD). Conclusion These cases may give us a possibility that intravenous CWGP offers potential benefits for non-small cell lung cancer patients.

Effect of Main Operating Conditions on Cathode Flooding Characteristics in a PEM Unit Fuel Cell (고분자전해질형 단위 연료전지의 주요 작동 조건이 공기극 플러딩 현상에 미치는 영향)

  • Min Kyoung-Doug;Kim Han-Sang
    • Transactions of the Korean Society of Mechanical Engineers B
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    • v.30 no.5 s.248
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    • pp.489-495
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    • 2006
  • Proton exchange membrane (PEM) should be sufficiently hydrated with a careful consideration of heat and water management. Water management has been a critical operation issue for better understanding the operation and optimizing the performance of a PEM fuel cell. The flooding on cathode side resulting from excess water can limit the fuel cell performance. In this study, the visual cell was designed and fabricated fur the visualization of liquid water droplet dynamics related to cathode flooding in flow channels. The experiment was carried out to observe the formation, growth and removal of water droplets using CCD imaging system. Effects of operating conditions such as cell temperature, air flow rate and air relative humidity on cathode flooding characteristics were mainly investigated. Based on this study, we can get the basic insight into flooding phenomena and its two-phase flow nature. It is expected that data obtained can be effectively used fur the setup and validation of two-phase PEM fuel cell models considering cathode flooding.

Effect of Radiation Dosage Changes on the Cell Viability and the Apoptosis Induction on Normal and Tumorigenic Cells (방사선의 선량변화가 수종의 정상세포와 종양세포주의 세포활성도와 apoptosis 유발에 미치는 영향)

  • Park In-Woo;Lee Sam-Sun;Heo Min-Suk;Choi Soon-Chul
    • Journal of Korean Academy of Oral and Maxillofacial Radiology
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    • v.29 no.2
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    • pp.435-449
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    • 1999
  • Purpose : The study was aimed to detect the differences in the cell viability and the apoptosis induction after irradiation on normal and tumorigenic cells. Materials and Methods : The study. that was generated for two human normal cells(RHEK, HGF-l) and two human tumor cells(KB. HT-1080). was tested using MTT assay at 1 day and 3 day after irradiation and TUNEL assay under confocal laser scanning microscope at 1 day after irradiation. Single irradiation of 0.5. 1, 2. 4. and 8Gy were applied to the cells. The two fractions of 1. 2. 4. and 8Gy were separated with a 4-hour time interval. The irradiation was done with 5.38Gy/min dose rate using Cs-137 irradiator at room temperature. Results and Conclusions : 1. In 3-day group. the cell viability of HGF-1 cell was significantly decreased at 2. 4 and 8Gy irradiation, the cell viability of KB cell was significantly decreased at 8Gy irradiation and the cell viability of HT-I080 cell was significantly decreased at 4 and 8Gy irradiation. 2. There was significant difference between RHEK and KB cell line in the cell viability of 3-day group at 8Gy irradiation. There was significant difference between RHEK and HGF-1 cell line in the cell viability of 3-day group at 4 and 8Gy irradiation. 3. There was a significantly decreased cell viability in 3-day group than those in 1-day group at 2. 4 and 8Gy on HGF-1 cell. at 4 and 8Gy on HT-I080 cell. at 8Gy on KB cell. 4. We could detect DNA fragmented cells only on KB cell. Number of apoptotic cells of KB cell was significantly increased at 4 and 8Gy irradiation. However, there was no correlation between cell viability and apoptosis. 5. On all 4 cell lines, there were no differences between single and split irradiation method in cell viability and apoptosis.

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