Mesenchymal stem cells (MSCs) are promising candidates for cell therapy and tissue engineering, but their application has been impeded by lack of knowledge of their core biological properties. In order to identify MSC-specific proteins, the hydrophobic protein fraction was individually prepared from two different umbilical cord blood (UCB)-derived MSC populations; these were then subjected to two-dimensional (2D) gel electrophoresis and peptide mass fingerprinting matrix-assisted laser desorption/ionization (MALDI)-time of flight (TOF)-mass spectrometry (MS). Although the 2D gel patterns differed somewhat between the two samples, computer-assisted image analysis identified shared protein spots. 35 spots were reliably identified corresponding to 32 different proteins, many of which were chaperones. Based on their primary sub-cellular locations the proteins could be grouped into 6 categories: extracellular, cell surface, endoplasmic reticular, mitochondrial, cytoplasmic and cytoskeletal proteins. This map of the water-insoluble proteome may provide valuable insights into the biology of the cell surface and other compartments of human MSCs.
Journal of Physiology & Pathology in Korean Medicine
/
v.27
no.1
/
pp.92-98
/
2013
Bee Venom(below BV) has been used in alternative medicine to treat the diseases, such as pain diseases. BV contains a variety of peptides, including melittin, apamin, adolapin, MCD peptide, enzymes(i.e. PLA2), amines(i.e. histamine and epinephrine), and nonpeptide components. The two main components of BV are melittin and PLA2. The cell cytotoxic effects through the activation of PLA2 by melittin have been suggested to be the critical mechanism for the depress of cancer cell. Melittin and PLA2 have been reported to induce apoptosis and to possess anti-cancer effects and neurite outgrowth in PC12 cells. Analysis of proliferation was confirmed by MTT assay. BV decreased cell number through dose- and duration-dependent manner and these effects are apparent at a concentration of 3 ${\mu}g/ml$. To observe which signaling molecules will be activated by BV, phosphorylation of ERK, p38 MAPK, JNK and ERM were examined by Western blot analysis. To study the long term effect of BV in human gastric adenocarcinoma cell lines, the image of cells treated with BV for 4 days were obtained. BV was shown to exhibit anti-cancer activity in human gastric adenocarcinoma cell lines at a broad range of concentrations of 3 ${\mu}g/ml$. ERK, p38 MAPK and JNK were found to increase in BV treated cells. However, ERM which known to be involved in the cell death, was gradually decreased to 30minutes after addition 3 ${\mu}g/ml$ of BV. These results provide a possible BV-induced inhibitory signal for cancer proliferation that is initiated by the decrease in ERM activity. Moreover, it is likely that the activation of ERK, p38 MAPK and JNK are required for the BV-induced inhibition of cancer proliferation.
Seo, Young-Rok;Han, Sung-Sik;Kim, L. O′Neill;Ryu, Jae-Chun
Proceedings of the Korea Society of Environmental Toocicology Conference
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1997.12a
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pp.101-101
/
1997
Comet assay, single cell gel electrophoresis has been known as useful, rapid, simple, visual, and sensitive technique for measuring the DNA breakage in mammalian ce1ls. For evaluation of DNA damage using comet assay, early studies reported a change in comet length and intensity with DNA damage using simple visual technique, such as fluorescence microscopy with eyespiece. In recent, some workers are observing and analyzing nucleotide of comets using quantitative fluorescence image analysis system to estimate 'tail moment', which is defined as the product of the tail length and the fraction of total DNA in tail. Our laboratory also adopted the image analysis software for qualification. In addition, many of the practical features of comet assay render it potentially attractive as useful tool for molecular toxicology and carcinogenesis, because the system is already showing considerable promise as rapid predictor in both in vitro and in vivo experimental designs. Recently, the comet assay becomes a attractive technique to study of apoptosis, because apoptotic fragmentation of nuclear DNA into nucleosomal sizes can be evaluated by the comet assay. So, we attempted to apply the comet assay to studying the effect of various stress on the apoptosis-sensitive cell lines. Particularly, focusing on the hyperthermic apoptosis, we could find that heat shock(44˚C for 60 minutes) was sufficient to induced apoptosis in these cell lines. But using the highly sensitive comet assay, we could not detect DNA breaks immediately after heat shock.
In molecular imaging studies via magnetic resonance imaging, in vivo cell tracking is an important issue for the observation of cell therapy or disease behavior. High resolution imaging and longitudinal study are necessary to track the cell movement. Since the field inhomogeneity extends over several voxels, we have performed the numerical analysis using the sub-voxel method dividing a voxel of MR image into several elements and the information about the field inhomogeneity distribution around the micro-beads. We imbedded ferrite-composite micro-beads with the size of $20-150{\mu}m$ in the subject substituted for cells to induce local field distortion. In the phantom imaging with the isotropic voxel size of $200{\mu}m^3$, we could confirm the feasibility of sub-voxel tracking in a 3.0 T MRI.
Kim Jeong-Hwa;Heo Min-Suk;Lee Sam-Sun;Choi Soon-Chul
Journal of Korean Academy of Oral and Maxillofacial Radiology
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v.29
no.1
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pp.87-103
/
1999
Purpose: This study was undertaken to quantitatively estimate the degree of the damage and recovery of the irradiated rat condylar cartilage using the Image Analyzer. Materials and Methods: Experimental animals were 16 male rats of the Sprague-Dawley strain at the age of 20 day irradiated with the dose of 10 Gy in their head and neck region. Four rats were sacrificed at the each of the following time intervals - 1, 4, 7 and 14 days, respectively. The same number of control group animals were sacrificed at the each age of 21. 24, 27 and 34 days, respectively. The specimens were stained with 0.5% toluidine blue and examined with light microscope. The condylar cartilage was divided into 4 zones; fibrous zone, proliferating zone, upper hypertrophic zone, and lower hypertrophic zone. And then, the proliferating zone was subdivided into 2 layers - upper and lower layer, and upper and lower hypertrophic zone were subdivided into three layers, respectively - upper, middle and lower layer. With the aid of Image Analyzer, morphometric analysis was performed. The thickness, the numerical density of cells, the cell area density, the extracellular matrix area density, the mean area of single cell, the mean area of extracellular matrix per single cell were measured and analysed. Results: In the experimental group, the thickness of the fibrous zone was slightly increased and that of the proliferating zone and the upper and the lower hypertrophic zone was markedly decreased. With time, the thickness of the fibrous zone was gradually increased and that of the proliferating zone and the upper and the lower hypertrophic zone was steadily in the decreased state. The numerical density of cells of the proliferating zone was increased on post-irradiated 1 day, but decreased after post-irradiated 4 day, and that of the upper hypertrophic zone was decreased. The numerical density of cells of the lower hypertrophic zone was decreased in the early stage and then was decreased or not significantly different from that of the control group with time. In the experimental group, the cell area density of the fibrous zone and the proliferating zone was decreased in the early stage and then gradually increased or not significantly different from that of the control group with time. The cell area density of the upper and the lower hypertrophic zone was varied with time. The extracellular matrix area density value were totally opposite to the cell area density values: The mean area of single cell of the fibrous zone and the proliferating zone was .decreased on post-irradiated 1 day, and increased after post-irradiated 4 day. The mean area of single cell of the upper hypertrophic zone was varied with each layer and time. In the experimental group, the mean area of extracellular matrix per single cell of the fibrous zone was not significantly different with control group, and that of the proliferating zone was decreased on post-irradiated 1 day, and increased after post-irradiated 4 day. The mean area of extracellular matrix per single cell of the lower hypertrophic zone was increased in the early stage. and that of upper hypertrophic zone was varied with each layer and time. Conclusion: The condylar cartilages of rats were affected by irradiation, but the changes were vaned with each layer and time. By morphometric analysis. the changes of the cells of the condylar cartilage of irradiated rat could be calculated quantitatively.
Hee-Jae Yoon;Si-Yoon Cho;Hyeong-Geug Kim;Ji-Yeon Lee
Journal of Pharmacopuncture
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v.27
no.2
/
pp.131-141
/
2024
Objectives: Polycystic ovary syndrome (PCOS) is one of the most common disorders and it shows up to 20% prevalence in reproductive-aged women populations, but no cures are available to date. We aimed to investigate the protective effects of Changbudodam-tang (CBD) on cell death signaling pathways, inflammation, and oxidative stress observed in Bone-Marrow derived human mesenchymal stem cell (BM-hMSC) by means of PCOS therapeutics in the future. Methods: BM-hMSCs were applied with cell deaths and injuries. Apoptosis and pyroptosis signals were quenched with their related signaling pathways using quantitative PCR, Western blot, and fluorescence image analysis. Results: Our data clearly displayed hydrogen peroxide- and nigericin-treated cell death signaling pathways via regulations of mitochondrial integrity and interleukin (IL)-1β at the cellular levels (p < 0.01 or 0.001). We further observed that pre-treatment with CBD showed protective effects against oxidative stress by enhancement of antioxidant components at the cellular level, with respect to both protein and mRNA expression levels (p < 0.05, 0.01 or 0.001). The mechanisms of CBD were examined by Western blot analysis, and it showed anti-cell death, anti-inflammatory, and antioxidant effects via normalizations of the Jun N-terminal kinase/mitogen-activated protein kinase kinase 7/c-Jun signaling pathways. Conclusion: This study confirmed the pharmacological properties of CBD by regulation of cellular oxidation and the inflammation-provoked cell death condition of BM-hMSCs, which is mediated by the MKK7/JNK/c-Jun signaling pathway.
Journal of the Korea Institute of Information and Communication Engineering
/
v.17
no.5
/
pp.1239-1244
/
2013
In the field of clinical medicine and research, the analysis of such as protein and DNA at the molecular level and even at the cell level are necessary for disease diagnosis and treatment. In many cases, a real time image of samples is needed for the accurate analysis and manipulation of samples since experimental samples are degenerated with time. In this research, a three-dimensional fluorescence microscope device was developed for taking images of protein and DNA inside a single cell and the server-client based image analysis system was made for an integrated management of the real-time images taken from the microscope device. The system consists of a fluorescent measurement device, the associated software and a client program on smartphone. The developed system allows doctors or experimental managers to receive and look at the real-time experimental images taken from the samples of patients anywhere in the emergency, to analyze results and to instantly diagnose the disease and to transfer the results to the patients. As a result, the system is able to be utilized in the implementation of ubiquitous health as well.
Kim Young-Jick;Lee Myung-Kon;Park Su-A;Shin Ho-Joon;Kim In-Ae;Lee Yong-Jae;Shin Ji-Won;Shin Jung-Woog
Proceedings of the Korean Society of Precision Engineering Conference
/
2005.10a
/
pp.69-72
/
2005
In this study, effects of IHPs with various resting times to cell adhesion were investigated through the measurements of cell adhesive force, number and area of focal contacts (stained vinculin spots), and projected cell area, perimeter and circularity. In addition correlation tests and curve estimations using the experimental results were performed fur the finding an essential factor for increment of cell adhesive force. Tn the results, immediately after mechanical stimuli (150 minutes after seeding) and one hour later (210 minutes after seeding), the average adhesive force of experimental group 5 (resting time: 15min) compared with that of control group at same culture time was increased significantly (p<0.05). Average projected area and perimeter of cells at Group 5 were increased significantly (p<0.05), while average circularity of cells at Group 5 incubated fur 210 minutes was decreased significantly (p<0.05). In the digital image analysis of focal contacts containing vinculins, area and numbers of focal contacts per cell at Group 5 were higher than those of the other groups. This study indicated that IHP with appropriate resting time could contribute in improving cell adhesive force, cell spreading, development of cytoskeleton and formation of focal contacts. And cell adhesive force was correlated to the morphological aspects of cell and development of focal contacts. Particularly, area of focal contacts was closely related to cell adhesive force.
Journal of the Korean Society for Precision Engineering
/
v.19
no.6
/
pp.136-144
/
2002
Melting method has long been considered difficult to realize because of problems such as the low foamability of molten metal, the varying size of cellular structures and solidification shrinkage. The parameters to solve the problem in electric furnace were stirring temperature, stirring velocity, heating velocity and foaming temperature It is important to consider the effects of induction heating, because it brings about the inner flow by the temperature gradient. Aspect ratio also depends on the induction heating. Mechanical properties are dependent on cell sizes and aspect rations. Therefore, this paper presents the effects of these parameters on the cell sizes. For the sake of this, combined stirring process was used to fabricate aluminum foam materials by the above mentioned parameters. Image analysis was performed to calculate the cell sizes, distributions, and aspect ratioes at the cross section of feared aluminum in the direction of height.
Journal of the Korea Institute of Information and Communication Engineering
/
v.4
no.5
/
pp.971-976
/
2000
The main objective of this paper is to present the rain cell size distribution observed during squall line episodes in the Sudano-Sahelian region. The used data were collected during the EPSAT Program [Etude des Precipitation par SATellite (Satellites Study of Precipitation)] which has been developed since 1958, on an experimental area located near Niamey, Niger (2 10′32"E, 13 28′38"N). The data were obtained with a C-band radar and a network composed of approximately 100 raingages over a 10,000 $\textrm{km}^2$. In this work a culling of the squall line episodes was made for the 1992 rainy season. After radar data calibration using the raingage network a number of PPI (Plan Position Indicator) images were generated. Each image was then treated in order to obtain a series of radar reflectivity (Z) maps. To describe the cell distribution, a contouring program was used to analyze the areas with rain rate greater than or equal to the contour threshold (R$\geq$$\tau$). 24700 contours were generated, where each iso-pleth belongs to a predefined threshold. Computing each cell surface and relating its area to an equi-circle (a circle having the same area as the cell), a statistical analysis was made. The results show that the number of rain cells having a given size is an inverse exponential function of the equivalent radius. The average and median equivalent radii ate 1.4 and 0.69 In respectively. Implications of these results for the precipitation estimation using threshold methods are discussed.
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