• Title/Summary/Keyword: Cell analysis

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Effect of Injin Butanol Fraction with Thin Layer Chromatography on Fas-mediated Apoptosis (인진butanol 분획의 TLC추출성분이 Fas-mediated Apoptosis에 미치는 영향)

  • 박용진;김영철;이장훈;우흥정
    • The Journal of Korean Medicine
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    • v.23 no.2
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    • pp.57-69
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    • 2002
  • Objective: The aim of this study is to investigate the effect of Injin butanol fractions with Thin Layer Chromatography on Fas-mediated Apoptosis. Method: Injin-butanol fraction separated by TLC. MIT assay, cell cycle analysis, Caspase-3 protease assay, DNA fragmentation assay and quantitative RT-PCR were performed to evaluate the effects of TLC extraction of lnjin-butanol fraction on cell viability, cell cycle progression and apoptosis. Results: Scopoletin, luteolin, apigenin and unknown powder was isolated by TLC. Fas-mediated apoptosis analysis shows that scopoletin has inhibiting function on apoptosis. Caspase- 3 protease assay analysis shows that scopoletin inhibits activity of caspase-3. Quantitative RT-PCR analysis shows that no activity on caspase-3, but apoptosis inhibition cytokine -Bcl-2- is activated, and apoptosis activating cytokine -Bax- is unactivated. Conclusion: These results show that each fraction of Injin-butanol TLC extraction, especially scopoletin, acts as a protective function on liver cell viability, and inhibitory function on apoptosis. (J Korean Oriental Moo 2002;23(2):57-69)

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Prediction of Mechanical and Electrical Properties of NiO-YSZ Anode Support for SOFC from Quantitative Analysis of Its Microstructure (미세조직 정량 분석을 통한 고체산화물연료전지용 NiO-YSZ 연료극 지지체의 기계적/전기적 성능 예측)

  • WAHYUDI, WANDI;KHAN, MUHAMMAD SHIRJEEL;SONG, RAK-HYUN;LEE, JONG-WON;LIM, TAK-HYOUNG;PARK, SEOK-JOO;LEE, SEUNG-BOK
    • Transactions of the Korean hydrogen and new energy society
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    • v.28 no.5
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    • pp.521-530
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    • 2017
  • Improving the microstructure of NiO/YSZ is one of several approaches used to enhance the electrical and mechanical properties of an anode support in Solid Oxide Fuel Cells (SOFCs). The aim of the work reported in this paper was to predict the relationship between these microstructural changes and the resulting properties. To this end, modification of the anode microstructure was carried out using different sizes of Poly (Methyl Methacrylate) (PMMA) beads as a pore former. The electrical conductivity and mechanical strength of these samples were measured using four-probe DC, and three-point bend-test methods, respectively. Thermal etching followed by high resolution SEM imaging was performed for sintered samples to distinguish between the three phases (NiO, YSZ, and pores). Recently developed image analysis techniques were modified and used to calculate the porosity and the contiguity of different phases of the anode support. Image analysis results were verified by comparison with the porosity values determined from mercury porosimetry measurements. Contiguity of the three phases was then compared with data from electrical and mechanical measurements. A linear relationship was obtained between the contiguity data determined from image analysis, and the electrical and mechanical properties found experimentally. Based upon these relationships we can predict the electrical and mechanical properties of SOFC support from the SEM images.

The Effect of Injin and Injinsaryungsangagambang on Liver Cell Viability, Lever Cell Cycle Progression and DNA Damage-induced Apoptosis (인진(茵蔯)과 인진사령산가감방(茵蔯四岺散加減方)이 간세포활성(肝細胞活性), 세포주기(細胞週期) 및 DNA damage-induced apoptosis에 미치는 영향(影響))

  • Kang, Woo-Sung;Lee, Jang-Hoon;Woo, Hong-Jung
    • The Journal of Korean Medicine
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    • v.20 no.1 s.37
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    • pp.91-105
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    • 1999
  • The effects of Yinjin and Yinjinsaryongsangagambang on a DNA damaging agent, etoposide-induced apoptosis, cell viability, cell cycle progression, and mRNA expression of apoptosis-related genes of human hepatocyte cell line HepG2 were investigated using tryphan blue exclusion assay, MTT assay, flow cytometry, immunocytometric analysis of PCNA, and quantitative RT-PCR analysis. MTT assay showed that Yinjin and Yinjinsaryongsangagambang increases cellular viability of HepG2 cells in a dosage-dependent manner. Stimulation of cell cycle progression by Yinjin or Yinjinsaryongsangagambang was detected by flow cytometric analysis of the DNA content and immunocytometric analysis of PCNA expression. A significant reduction of a DNA-damaging agent, etoposide-induced apoptosis were found in both Yinjin and Yinjinsaryongsangagambang-treated cells in dosage-dependent manner. In overall, 3-fold reduction of apoptosis was recognized in $10.0\;{\mu}g/ml$ of Yinjin or Yinjinsaryongsangagambang-treated cells compared to untreated cells. Although the difference is not significant, Yinjinsaryongsangagambang showed slightly higher effect on the inhibition of apoptosis than Yinjin. From flow cytometric analysis of apoptosis, while 39.9% of untreated cells showed etoposide-induced apoptotic cell death, only 19.6% or 17.4% of Yinjin or Yinjinsaryongsangagambang-treated cells were fond at apoptotic sub G1 phase, respectively. Interestingly, strong induction of Gadd45-mRNA was observed from Yinjin or Yinjinsaryongsangagambang-treated cells. However, no changes in expression levels of p53 and Waf1 were detected, demonstrating that induction of Gadd45 mRNA expression by Yinjin or Yinjinsaryongsangagambang occurs by p53-independent mechanism. Marked mRNA inductions of two apoptosis-inhibiting genes, Bcl-2 and Bcl- XL, were found in both Yinjin or Yinjinsaryongsangagambang-treated HepG2 cells while no changes was detected in expression levels of an apoptosis-promoting gene, Bax.

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DNA METHYLATION OF TPEF GENE IN HEAD AND NECK SQUAMOUS CELL CARCINOMA CELL LINES (두경부암 세포주에서 TPEF 유전자의 methylation 변이)

  • Chun, So-Young;Kim, Jung-Ock;Hong, Su-Hyung;Chung, Yu-Kyung;Jang, Hyun-Jung;Shon, Yoon-Kyung;Kim, Jung-Wan
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.31 no.6
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    • pp.468-473
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    • 2005
  • Head and neck squamous cell carcinoma (HNSCC) is the sixth most common malignancy worldwide. The molecular mechanisms involved in the development and progression of these carcinomas are not well known. Abnormalities of genomic methylation patterns have been attributed a role in carcinogenesis and local de novo methylation at tumor suppressor loci was held to be involved in silencing of tumor suppressor genes. Using Ms APPCR, we previously isolated a hypermethylated fragment corresponded to the 5' end of TPEF gene from primary liver and lung cancer cells. To confirm the inactivation of TPEF gene by hypermethylation in HNSCC, we investigated correlation between methylation pattern and expression of TPEF in 10 HNSCC cell lines. In methylation analysis such as combined-bisulfite restriction analysis(COBRA) and bisulfite sequencing, only RPMI 2650 showed none methylated pattern and another 9 cell lines showed dense methylation. The TPEF gene expression level analysis using RT-PCR showed that these 9 cell lines had not or significantly low expression levels of TPEF as compared with RPMI 2650. In addition, the increase of TPEF reexpression by 5-AzaC as demethylating agent in 9 cell lines also indicated that TPEF expression was regulated by hypermethylation. These results of this study demonstrate that epigenetic silencing of TPEF gene by aberrant methylation could play an important role in HNSCC carcinogenesis.

Automated Bacterial Cell Counting Method in a Droplet Using ImageJ (이미지 분석 프로그램을 이용한 액적 내 세포 계수 방법)

  • Jingyeong Kim;Jae Seong Kim;Chang-Soo Lee
    • Korean Chemical Engineering Research
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    • v.61 no.2
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    • pp.247-257
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    • 2023
  • Precise counting of cell number stands in important position within clinical and research laboratories. Conventional methods such as hemocytometer, migration/invasion assay, or automated cell counters have limited in analytical time, cost, and accuracy., which needs an alternative way with time-efficient in-situ approach to broaden the application avenue. Here, we present simple coding-based cell counting method using image analysis tool, freely available image software (ImageJ). Firstly, we encapsulated RFP-expressing bacteria in a droplet using microfluidic device and automatically performed fluorescence image-based analysis for the quantification of cell numbers. Also, time-lapse images were captured for tracking the change of cell numbers in a droplet containing different concentrations of antibiotics. This study confirms that our approach is approximately 15 times faster and provides more accurate number of cells in a droplet than the external analysis program method. We envision that it can be used to the development of high-throughput image-based cell counting analysis.

ROLE OF DCC(DELETED IN COLORECTAL CANCER) GENE IN ORAL SQUAMOUS CELL CARCINOMA (구강편평상피암종에서 DCC 유전자의 역할)

  • Ko, Seong-Kyu;Han, Se-Jin;Kim, Kyung-Wook
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.34 no.5
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    • pp.518-524
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    • 2008
  • Chromosome 18q alteration plays a key role in colorectal tumorigenesis, and loss of heterozygosity at 18q is associated with a poor prognosis in colon cancer. DCC(Deleted in Colorectal Cancer) is a putative tumor- suppressor gene at 18q21 that encodes a transmembrane protein with structural similarity to neural cell adhesion molecule that is involved in both epithelial and neuronal cell differentiation. DCC is implicated in regulation of cell growth, survival and proliferation. Thus, tumor progression in squamous cell carcinoma, stomach cancer, colorectal cancer correlates with downregulation of DCC expression. The mechanism for DCC suppression is associated with hypermethylation of the DCC gene promoter region. Hence, the goal of this study is to identify the promoter methylation responsible for the down-regulation of DCC expression in oral squamous cell carcinoma. 12 of tissue specimens for the study are excised and gathered from 12 patients who are diagnosed as SCC in department of OMS, dental hospital, dankook university. To find expression of DCC in each tissue samples, immunohistochemical staining, RT-PCR gene analysis and methylation specific PCR are processed. The results are as follows. 1. In the DCC gene RT-PCR analysis, 5(41.6%) of 12 specimens of oral squamous cell carcinoma did not expressed DCC gene. 2. In the promoter methylation specific PCR analysis, 5(41.6%) of 12 specimens showed promoter methylation of DCC gene. 3. In the immunohistochemical staining of poor differentiated and invasive oral squamous cell carcinoma, loss of DCC expression was observed. These findings suggest that methylation of the DCC gene may play a role in loss of gene expression in invasive oral squamous cell carcinoma.

Development of EIS Evaluation Method about PEMFC 1kW STACK (가정용 연료전지 스택의 EIS 평가 기법 개발)

  • Park, Chaneom;Han, Woonki;Jung, Jinsu;Ko, Wonsik
    • 한국신재생에너지학회:학술대회논문집
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    • 2011.05a
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    • pp.100.1-100.1
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    • 2011
  • Electrochemical impedance spectroscopy(EIS) are using widely as a useful technique mainly in the field of electrochemical for the analysis of electrode reactions or characteristics of the composites. The response analysis of the systems technique provides comprehensive informations about the characteristic and structure of complex and internal reaction. The EIS is the method to measure impedance of the measurement target classified by the frequency, it select the equivalent impedance model to give same response from the result and it calculate the parameter. Therefore, the chemical reaction inside the fuel cell is to modeling to electrical impedance. And as repeating the same experiment in each of the operating point, we can get each different parameter. As a result, we can establish the equivalent impedance model in each operating point. Therefore, if we use these models, we can evaluate the fuel cell without the internal design parameter of the fuel cell as required in existing modeling. The EIS is used typically technique for distinguish status of fuel cell called SOH(State Of Health). When the fuel cell is degradation, Efficiency and health of the fuel cell is reduced because internal impedance is increase. As usage of these principles, we can evaluate state of fuel cell through the impedance analysis of fuel cells. In this study, we are presents EIS distinction system and algorithm for residential fuel cell systems. At the time of the fuel cell installation in the fields, the EIS system and proposed algorithm will be able to apply as technique for efficiency and performance evaluation about fuel cell system.

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Inhibition of Cell-Cycle Progression in Human Promyelocytic Leukemia HL-60 Cells by MCS-C2, Novel Cyclin-Dependent Kinase Inhibitor

  • Kim, Min-Kyoung;Cho, Youl-Hee;Kim, Jung-Mogg;Chun, Moon-Woo;Lee, Seung-Ki;Lim, Yoong-Ho;Lee, Chul-Hoon
    • Journal of Microbiology and Biotechnology
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    • v.13 no.4
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    • pp.607-612
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    • 2003
  • To elucidate the action mechanism of MCS-C2, a novel analogue of toyocamycin and sangivamycin, its effect on the expression of cell cycle-related proteins in the human myelocytic leukemia cell line HL-60 was examined using Western blotting and a flow cytometric analysis. MCS-C2, a selective inhibitor of cyclin-dependent kinases, was found to inhibit cell growth in a time- and dose-dependent manner, and inhibits cell cycle progression by inducing the arrest at G1 and G2/M phases, in HL-60 cells. The flow cytometric analysis revealed an appreciable arrest of cells in the G2/M phase of the cell cycle after treatment with MCS-C2. The HL-60 cell population increased gradually from 13% at 0 h, to 28% at 12 h in the G2/M phase, after exposure to $2{\;}\mu\textrm{M}$ MCS-C2. Furthermore, Western blot analysis demonstrated that MCS-C2 induced the cell cycle arrest at G1 phase through the inhibition of pRb phosphorylation. Hypophosphorylated pRb accumulated after treatment with $5{\;}\mu\textrm{M}$ MCS-C2 for 12 h, whereas, the level of hyperphosphorylated pRb was reduced. Thus, treatment of the cell with MCS-C2 suppressed the hyperphosphorylated form of pRb with a commensurate increase in the hypophosphorylated form.

Effect of Hyulbuchukeotang on the Inhibition of Proliferation of Uterine leiomyoma cells and Cell apoptosis (혈부축어탕(血府逐瘀湯) 자궁근종세포의 증식억제와 Apoptosis 관련 유전자 발현에 미치는 영향)

  • Moon, Na-Young;Baek, Seung-Hee;Kim, Dong-Chul
    • The Journal of Korean Obstetrics and Gynecology
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    • v.19 no.2
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    • pp.186-198
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    • 2006
  • Purpose : The purpose of this study is to demonstrate the direct inhibitory effect of Hyulbuchukeotang on the proliferation of uterine leiomyoma cells through an experiment treating uterine leiomyoma cells cultivated by explantation with indicated concentrations of Hyulbuchukeotang and to research the gene expression related to cell cycle ill order to discover the connection with apoptosis and its mechanism by analyzing cell cycle. Methods : After primary culture of uterine leiomyoma cells, the cultivated uterine leiomyoma cells were treated with indicated concentrations of Hyulbuchukeotang for 24 hours. The inhibitory effect on the cell proliferation was determined by the cell count assay. The value of a cell count assay represent the percentage of cells in a phase of the cell cycle compared with total cells. In addition, a link between Hyulbuchukeotang and apoptosis was examined through flow cytometric analysis by FACS and DNA fragmentation analysis. Finally, the degree of gene expression related to cell cycle was evaluated by Western blot analysis. Results : The inhibitory effect of Hyulbuchukeotang increase of uterine leiomyoma cells treated with indicated concentrations of Hyulbuchkeotang increases. The result of gene expression related to G1 phase after treating with 100, 250, 500, 1,000 ${\mu}g/ml$ concentrations of Hyulbuchukeotang. on uterine leiomyoma cells is that the gene expression of p27 was increased but that of p53 an p21 remained unchanged and the gene of pRB, pro-caspase 3 was decreased. Conclusion Through the mentioned experiments, it is demonstrated that Hyulbuchkeotang is effective in inhibiting Proliferation of uterine leiomyoma cells by extending cell cycle G1. However it is not considered that the inhibitory effect results from the aptoposis.

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Evacuation Analysis for a Passenger Ship Using a Cellular Automata Model with Group Behavior (집단 행동을 나타내는 Cellular Automata 모델을 사용한 여객선 승객 탈출 분석)

  • Cha, Ju-Hwan;Ha, Sol;Lee, Kyu-Yeul
    • Journal of the Korea Society for Simulation
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    • v.20 no.4
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    • pp.149-155
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    • 2011
  • This paper presents simulations of advance evacuation analysis using a cellular automata model for passenger behavior in an emergency of passenger ship. The proposed cellular automata model divides the space in a uniform grid called "cell." Each passenger is located in a cell and moves to another cell according to a set of local rules assumed to be associated with the individual and crowd behaviors of the passengers. To verify the usefulness of the proposed cellular automata model, 11 tests, all of which are specified in International Maritime Organization Maritime Safety Committee/ Circulation 1238 (IMO MSC/Circ. 1238), were implemented, and it was confirmed that all the requirements of these tests had been met.