• Title/Summary/Keyword: FACS

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Induced apoptosis in human Uterine Leiomyoma Cells by treatment with Chiljehyangbu-hwan (칠제향부환(七製香附丸)이 자궁근종세포의 성장억제와 세포자멸사에 미치는 영향)

  • Kim, Seuk-Jung;Beak, Seung-Hee;Kim, En-Ha;Kim, Dong-Chul
    • The Journal of Korean Obstetrics and Gynecology
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    • v.20 no.2
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    • pp.25-42
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    • 2007
  • Purpose : Uterine leiomyoma (fibroids) are benign smooth muscle tumors originating from the myometrium. These benign neoplasms of monoclonal origin are typically diagnosed during the reproductive years, occurring only after puberty and tending to regress after menopause. In the present study we used Chiljehyangbu-hwan to determine its growth inhibitory effect and apoptosis in human uterine leiomyoma cells. Methods : Primary cultured human uterine leiomyoma cells were treated with Chiljehyangbu-hwan. Cell viability analysis was analyzed by MTS assay and FACS was performed to ascertain the effects Chiljehyangbu-hwan. DNA fragmentation analysis and casapase-3 activity test were done. Expression of apoptosis related proteins were evaluated by Western blot analysis. Results : Cell viability was significantly influenced by Chiljehyangbu-hwan treatment in a dose-dependent manner in leiomyoma cells compare to normal myometrial cells. FACS showed that Chiljehyangbu-hwan induced Sub G1 arrest. DNA fragmentation assay was carried out and apoptosis was detected. Activation of caspase-3, down-regulation of Bcl-2, with concomitant increased expression in Bid and Bax were observed. Chiljehyangbu-hwan treatment of uterine leiomyoma cells resulted in a concentration-dependent cell death induced via the mitochondrial pathway.

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A Trial of Screening of Genes Involved in Odontoblasts Differentiation from Human Dental Pulp Stem Cells

  • Park, Yoon-Kyu;Kim, Hyun-Jin
    • International Journal of Oral Biology
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    • v.37 no.4
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    • pp.167-173
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    • 2012
  • This study investigated the genes involved in the differentiation of odontoblasts derived from human dental pulp stem cells (hDPSCs). hDPSCs isolated from human tooth pulp were validated by fluorescence activated cell sorting (FACS). After odontogenic induction, hDPSCs were analyzed investigated by Alizaline red-S staining, ALP assay, ALP staining and RT-PCR. Differential display-polymerase chain reaction (DD-PCR) was performed to screen differentially expressed genes involved in the differentiation of hDPSCs. By FACS analysis, the stem cell markers CD24 and CD44 were found to be highly expressed in hDPSCs. When hDPSCs were treated with agents such as ${\beta}$-glycerophosphate (${\beta}$-GP) and ascorbic acid (AA), nodule formation was exhibited within six weeks. The ALP activity of hDPSCs was found to elevate over time, with a detectable up-regulation at 14 days after odontogenic induction. RT-PCR analysis revealed that dentin sialophosphoprotein (DSPP) and osteocalcin (OC) expression had increased in a time-dependent manner in the induction culture. Through the use of DD-PCR, several genes were differentially detected following the odontogenic induction. These results suggest that these genes may possibly be linked to a variety of cellular process during odontogenesis. Furthermore, the characterization of these regulated genes during odontogenic induction will likely provide valuable new insights into the functions of odontoblasts.

Protective Effect of Oak Extracts on Oxidative Stress Induced by Hydrogen Peroxide

  • Nam, Jeong Bin;Park, Hyung Bin;Jung, Ji Young;Yang, Jae-Kyung
    • Journal of the Korean Wood Science and Technology
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    • v.42 no.5
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    • pp.523-532
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    • 2014
  • This study was done to evaluate the antioxidant effect of oak hot water extracts on the oxidative stress induced by reactive oxygen species (ROS). The cytotoxicity of $H_2O_2$-induced oxidative stress was performed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay for the cell viability according to the dose-dependent treatment. Oak extracts demonstrated a dose-dependent ability to inhibit $H_2O_2$-induced apoptosis in cultured tenofibroblasts, as assessed by MTT assay and FACS analysis. $H_2O_2$ increased the phosphorylation of extracellular regulated kinase1/2 (ERK1/2) and of c-Jun N-terminal kinase (JNK) and the production of reactive oxygen species (ROS). In contrast, treatment with oak extracts was decreased this activation of ERK1/2 and JNK, as confirmed by western blot analysis, and reduced the production of ROS, as verified by fluorescent microscopic and flow cytometry (FACS) analyses. These findings suggest that oak extracts, by suppressing JNK, ERK1/2, and intracellular ROS production, have a concentration-dependent antiapoptotic effect on achilles tenofibroblasts exposed to an oxidative stressor, and may have therapeutic potential.

Facial Behavior Recognition for Driver's Fatigue Detection (운전자 피로 감지를 위한 얼굴 동작 인식)

  • Park, Ho-Sik;Bae, Cheol-Soo
    • The Journal of Korean Institute of Communications and Information Sciences
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    • v.35 no.9C
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    • pp.756-760
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    • 2010
  • This paper is proposed to an novel facial behavior recognition system for driver's fatigue detection. Facial behavior is shown in various facial feature such as head expression, head pose, gaze, wrinkles. But it is very difficult to clearly discriminate a certain behavior by the obtained facial feature. Because, the behavior of a person is complicated and the face representing behavior is vague in providing enough information. The proposed system for facial behavior recognition first performs detection facial feature such as eye tracking, facial feature tracking, furrow detection, head orientation estimation, head motion detection and indicates the obtained feature by AU of FACS. On the basis of the obtained AU, it infers probability each state occur through Bayesian network.

The Importance of FACS Analysis in the Development of Aptamers Specific to Pathogens

  • Moon, Ji-Hea;Kim, Giyoung;Park, Saet Byeol;Lim, Jongguk;Mo, Changyeun
    • Journal of Biosystems Engineering
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    • v.39 no.2
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    • pp.111-114
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    • 2014
  • Purpose: This review aims to introduce aptamers and the methods of its development to improve the sensitivity and selectivity to target bacteria. In this review, we have highlighted current developments and directions in the pathogen detection based on aptamers. Background: Aptamers, the specific nucleic acid sequences, can bind to targets with high affinity and specificity. Some of researches on the use of aptamers for the detection of pathogen have been reported in recent years. Aptamers have more applicability than antibodies for the development of pathogen detection using biosensor; such as easy to synthesis and labeling, lack of immunogenicity, and a low cost of production. However, only few reports on the development and use of aptamers for the detection of pathogen have been published. Review: Aptamers specific to pathogen are obtained by whole-cell systematic evolution of ligands by exponential enrichment (SELEX) process. SELEX process is composed of screening random oligonucleotide bound with target cells, multiple separation and amplification of nucleic acids, final identification of the best sequences. For improving those affinity and selectivity to target bacteria, optimization of multiple separating process to remove unbounded oligonucleotides from aptamer candidates and sorting process by flow cytometry are required.

Neuroprotective Effects of Some Plant Extracts Against Dopamine-induced Oxidative Stress on Neuronal Cell (Dopamine에 의해 산화적 스트레스를 받은 Neuronal Cell에 뇌 보호 효과를 가지는 수종 생약추출물의 검색)

  • Koo, Uk;Lee, Hak-Ju;Lee, Dong-Ho;Lee, Hyun-Jung;Ham, Ah-Rom;Mar, Woong-Chon
    • Korean Journal of Pharmacognosy
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    • v.40 no.1
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    • pp.41-45
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    • 2009
  • Parkinson's disease (PD) is the second most common neurodegenerative disorder after Alzhemier's disease. Neuropathologically, PD is characterized by the selective loss of dopaminergic neurons. The neuronal toxicity of cytosolic excess dopamine (DA) has been described in many studies using several cell lines. In dopaminergic neurons, cytosolic excess DA is easily oxidized via monoamine oxidase (MAO)-B, tyrosinase or by auto-oxidation to produce neurotoxic metabolites such as DA quinone. So, in the present study, we induced cell death by treatment of DA ($600{\mu}M$) in human neuroblastoma SH-SY5Y cell which was treated samples before 24 hr, and cell viability was measured by fluorescence activated cell sorter (FACs) analysis. Of those tested, the extracts of Poria cocos (赤茯笭)(whole), Gastrodia elata (rhizomes), Eucommia ulmoides (炒)(barks), Syneilesis palmata (whole), Acorus gramineus (rhizomes), Ligustrum japonicum (leaves) showed neuroprotective effects in dose dependent manner.

Protective Effect of NACA on Periodontal Stem Cell (NACA 처리에 따른 치주줄기세포 사멸 억제 효과)

  • Lee, Kyunghee
    • Journal of The Korean Society of Integrative Medicine
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    • v.8 no.3
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    • pp.53-62
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    • 2020
  • Purpose :Periodontal ligament stem cells maintain tissue homeostasis in periodontal ligament. The purpose of this study was to determine the characteristics of periodontal ligament stem cells isolated from premolar teeth and observe protective effects against oxidative damage caused by Triethylene glycol dimethacrylate (TEGDMA) following treatment with N-acetylsysteine amide (NACA) drug known as enzymatic antioxidants. Methods : Primary periodontal ligament stem cell (PDSC) culture was performed from simply extracted human premolar of orthodontic patients. The characteristics of the primary cultured PDSCs was analyzed using the FACS system. PDSCs was incubated with TEGDMA and NACA. The cell proliferation and survival was determined using WST-1 assay. Collected data were analyzed using SPSS Window 20. Results : Primary cultured PDSCs grow on the floor and develop rapidly in a cluster form from up to 14 days. The morphology of PDSCs showed the spindle-shaped cells and grew directionally. FACS analysis, In addition, positive expression of visible cells were observed in mesenchymal stem cell biomarkers. PDLSCs cell viability was significantly decreased at high concentration in both 3 and 6 hours after TEGDMA treatment. We observed a decrease in the number of cells as well as a morphological change of PDLSCs. Antioxidative effect was notable since the death of PDLSC death was significantly inhibited compared to the control group at 24 and 48 hours after NACA treatment. Conclusion : Therefore, based on the results of this study, further research should be encouraged considering the development of clinical treatment methods using various antioxidants as well as regenerative engineering techniques utilizing periodontal ligament stem cells.

Effects of Different Lightweight Functional Fillers for Use in Cementitious Composites

  • Hanif, Asad;Lu, Zeyu;Cheng, Yu;Diao, Su;Li, Zongjin
    • International Journal of Concrete Structures and Materials
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    • v.11 no.1
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    • pp.99-113
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    • 2017
  • The effects of different lightweight functional fillers on the properties of cement-based composites are investigated in this study. The fillers include fly ash cenospheres (FACs) and glass micro-spheres (GMS15 and GMS38) in various proportions. The developed composites were tested for compressive, flexural and tensile strengths at 10 and 28-day ages. The results indicated that both FACs and GMS38 are excellent candidates for producing strong lightweight composites. However, incorporation of GMS15 resulted in much lower specific strength values (only up to $13.64kPa/kg\;m^3$) due to its thinner shell thickness and lower isostatic crushing strength value (2.07 MPa). Microstructural analyses further revealed that GMS38 and GMS15 were better suited for thermal insulating applications. However, higher weight fraction of the fillers in composites leads to increased porosity which might be detrimental to their strength development.

Simulation of a Six-zone Simulated Moving Bed Chromatographic Process for NPK Fertilizer Production (NPK 비료 생산을 위한 6구역 모사이동층의 공정모사)

  • Lim, Youngil;Lee, Aran
    • Korean Chemical Engineering Research
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    • v.45 no.1
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    • pp.1-11
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    • 2007
  • Using strong cation exchange simulated moving bed (SMB) chromatography, a nitrogen-phosphate-potassium (NPK) fertilizer is produced in a cost-effective manner. The SMB process operated in a non-traditional way is divided into production and regeneration sections for exclusion of undesirable ions, and composed of six zones including two wash-water zones. This paper addresses modeling and simulation studies on the ion-exchange SMB process, comparing simulation results with experimental data obtained both from a pilot plant and an industrial plant. The simulation results show a good agreement with in situ experimental data obtained in the two plants. The model equation validated by the experiments will be applicable for optimization problems to obtain optimum operating conditions of the process.

Cell Surface Expression of Tumor Necrosis Factor-Alpha by Activated Rat Astrocytes

  • Chung, Il-Yup;Benveniste, Etty N.
    • BMB Reports
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    • v.29 no.6
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    • pp.530-534
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    • 1996
  • Astrocyte are the major glial cell type in the central nervous system (CNS), and analogous to macrophage, mediates the number of immune responses such as production of cytokines including tumor necrosis factor alpha ($TNF-{\alpha}$) upon activation. $TNF-{\alpha}$ has been implicated in neuroimmunological disorders through killing oligodendrocytes and thus causing demyelination. It has been previously demonstrated that mitogen-activated T cells synthesized a 26 kDa precursor form of $TNF-{\alpha}$ which is bound to the surface of a membrane, and is later secreted as a 17 kDa mature version. In order to examine whether astrocytes would produce the transmembrane form of $TNF-{\alpha}$, astrocytes were stimulated with biological stimuli and the membrane form of $TNF-{\alpha}$ was analyzed by Western blot and FACS analysis. When astrocytes are stimulated with lipopolysaccharide (LPS), $IFN-{\gamma}/LPS$, or $IFN-{\gamma}/IL-1{\beta}$, they were able to express a membrane-anchored $TNF-{\alpha}$ of approximately 26 kDa protein which was immunoreactive to an $anti-TNF-{\alpha}$ antibody, whereas unstimulated astrocytes or astrocytes treated with $IFN-{\gamma}$ or $IL-1{\beta}$ alone was not. Our FACS data were also consistent with the immunoblot analysis. Our result suggests that the membrane form of $TNF-{\alpha}$ expressed by activated astrocytes may cause local damage to oligodendrocytes by direct cell-cell contact and contribute to demyelination observed in multiple sclerosis (MS) and experimental allergic encephalomyelitis (EAE).

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