Search results for: regulatory T cell
4150 Modeling of Oxygen Supply Profiles in Stirred-Tank Aggregated Stem Cells Cultivation Process
Authors: Vytautas Galvanauskas, Vykantas Grincas, Rimvydas Simutis
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This paper investigates a possible practical solution for reasonable oxygen supply during the pluripotent stem cells expansion processes, where the stem cells propagate as aggregates in stirred-suspension bioreactors. Low glucose and low oxygen concentrations are preferred for efficient proliferation of pluripotent stem cells. However, strong oxygen limitation, especially inside of cell aggregates, can lead to cell starvation and death. In this research, the oxygen concentration profile inside of stem cell aggregates in a stem cell expansion process was predicted using a modified oxygen diffusion model. This profile can be realized during the stem cells cultivation process by manipulating the oxygen concentration in inlet gas or inlet gas flow. The proposed approach is relatively simple and may be attractive for installation in a real pluripotent stem cell expansion processes.Keywords: aggregated stem cells, dissolved oxygen profiles, modeling, stirred-tank, 3D expansion
Procedia PDF Downloads 3004149 Legal Regulation of Personal Information Data Transmission Risk Assessment: A Case Study of the EU’s DPIA
Authors: Cai Qianyi
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In the midst of global digital revolution, the flow of data poses security threats that call China's existing legislative framework for protecting personal information into question. As a preliminary procedure for risk analysis and prevention, the risk assessment of personal data transmission lacks detailed guidelines for support. Existing provisions reveal unclear responsibilities for network operators and weakened rights for data subjects. Furthermore, the regulatory system's weak operability and a lack of industry self-regulation heighten data transmission hazards. This paper aims to compare the regulatory pathways for data information transmission risks between China and Europe from a legal framework and content perspective. It draws on the “Data Protection Impact Assessment Guidelines” to empower multiple stakeholders, including data processors, controllers, and subjects, while also defining obligations. In conclusion, this paper intends to solve China's digital security shortcomings by developing a more mature regulatory framework and industry self-regulation mechanisms, resulting in a win-win situation for personal data protection and the development of the digital economy.Keywords: personal information data transmission, risk assessment, DPIA, internet service provider, personal information data transimission, risk assessment
Procedia PDF Downloads 524148 Fabrication of Silicon Solar Cells Using All Sputtering Process
Authors: Ching-Hua Li, Sheng-Hui Chen
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Sputtering is a popular technique with many advantages for thin film deposition. To fabricate a hydrogenated silicon thin film using sputtering process for solar cell applications, the ion bombardment during sputtering will generate microstructures (voids and columnar structures) to form silicon dihydride bodings as defects. The properties of heterojunction silicon solar cells were studied by using boron grains and silicon-boron targets. Finally, an 11.7% efficiency of solar cell was achieved by using all sputtering process.Keywords: solar cell, sputtering process, pvd, alloy target
Procedia PDF Downloads 5754147 User-Friendly Task Creation Using a CAD Integrated Robotic System on a Real Workcell
Authors: Alireza Changizi, Arash Rezaei, Jamal Muhammad, Jyrki Latokartano, Minna Lanz
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Offline programming (OLP) is a new method in robot programming which is used widely in the industry nowadays which is a simulation base method that can produce the robot codes for motion according to virtual world in the simulation software. In this project Delmia v5 is used as simulation software. First the work cell component was modelled by Catia v5 and all of them was imported to a process file in Delmia and placed roughly to form the virtual work cell. Then robot was added to the work cell from the Delmia library. Work cell was calibrated corresponding to real world work cell to have accurate code. Tool calibration is the first step of calibration scheme and then work cell equipment can be calibrated using 6 point calibration method. Finally generated code needs to be reformed to match related controller code instruction. At the last stage IO were set to accomplish robots cooperation and make their motion synchronized. The pros and cons also will be discussed to clarify the presented results show the feasibility of the method and its effect on production line efficiency. Finally the positive and negative points of the implementation will be discussed.Keywords: robotic, automated, production, offline programming, CAD
Procedia PDF Downloads 3824146 Characterization of a LiFeOP₄ Battery Cell with Mechanical Responses
Authors: Ki-Yong Oh, Eunji Kwak, Due Su Son, Siheon Jung
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A pouch type of 10 Ah LiFePO₄ battery cell is characterized with two mechanical responses: swelling and bulk force. Both responses vary upon the state of charge significantly, whereas voltage shows flat responses, suggesting that mechanical responses can become a sensitive gauge to characterize microstructure transformation of a battery cell. The derivative of swelling s with respect to capacity Q, (ds/dQ) and the derivative of force F with respect to capacity Q, (dF/dQ) more clearly identify phase transitions of cathode and anode electrodes in the overall charge process than the derivative of voltage V with respect to capacity Q, (dV/dQ). Especially, the force versus swelling curves over the state of charge clearly elucidates three different stiffness over the state of charge oriented from phase transitions: the α-phase, the β-phase, and the metastable solid-solution phase. The observation from mechanical responses suggests that macro-scale mechanical responses of a battery cell are directly correlated to microscopic transformation of a battery cell.Keywords: force response, LiFePO₄ battery, strain response, stress response, swelling response
Procedia PDF Downloads 1654145 Numerical Study on Response of Polymer Electrolyte Fuel Cell (PEFCs) with Defects under Different Load Conditions
Authors: Muhammad Faizan Chinannai, Jaeseung Lee, Mohamed Hassan Gundu, Hyunchul Ju
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Fuel cell is known to be an effective renewable energy resource which is commercializing in the present era. It is really important to know about the improvement in performance even when the system faces some defects. This study was carried out to analyze the performance of the Polymer electrolyte fuel cell (PEFCs) under different operating conditions such as current density, relative humidity and Pt loadings considering defects with load changes. The purpose of this study is to analyze the response of the fuel cell system with defects in Balance of Plants (BOPs) and catalyst layer (CL) degradation by maintaining the coolant flow rate as such to preserve the cell temperature at the required level. Multi-Scale Simulation of 3D two-phase PEFC model with coolant was carried out under different load conditions. For detailed analysis and performance comparison, extensive contours of temperature, current density, water content, and relative humidity are provided. The simulation results of the different cases are compared with the reference data. Hence the response of the fuel cell stack with defects in BOP and CL degradations can be analyzed by the temperature difference between the coolant outlet and membrane electrode assembly. The results showed that the Failure of the humidifier increases High-Frequency Resistance (HFR), air flow defects and CL degradation results in the non-uniformity of current density distribution and high cathode activation overpotential, respectively.Keywords: PEM fuel cell, fuel cell modeling, performance analysis, BOP components, current density distribution, degradation
Procedia PDF Downloads 2094144 Optimization of Heterojunction Solar Cell Using AMPS-1D
Authors: Benmoussa Dennai, H. Benslimane, A. Helmaoui
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Photovoltaic conversion is the direct conversion of electromagnetic energy into electrical energy continuously. This electromagnetic energy is the most solar radiation. In this work we performed a computer modelling using AMPS 1D optimization of hetero-junction solar cells GaInP / GaAs configuration for p / n. We studied the influence of the thickness the base layer in the cell offers on the open circuit voltage, the short circuit current and efficiency.Keywords: optimization, photovoltaic cell, GaInP / GaAs AMPS-1D, hetetro-junction
Procedia PDF Downloads 5134143 Energy Management of Hybrid Energy Source Composed of a Fuel Cell and Supercapacitor for an Electric Vehicle
Authors: Mejri Achref
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This paper proposes an energy management strategy for an electrical hybrid vehicle which is composed of a Proton Exchange Membrane (PEM) fuel cell and a supercapacitor storage device. In this paper, the mathematical model for the proposed power train, comprising the PEM Fuel Cell, supercapacitor, boost converter, inverter, and vehicular structure, was modeled in MATLAB/Simulink. The proposed algorithm is evaluated for the Highway Fuel Economy Test (HWFET) driving cycle. The obtained results demonstrate the effectiveness of the proposed energy management strategy in reduction of hydrogen consumption.Keywords: proton exchange membrane fuel cell, hybrid vehicle, hydrogen consumption, energy management strategy
Procedia PDF Downloads 1734142 Functional Switching of Serratia marcescens Transcriptional Regulator from Activator to Inhibitor of Quorum Sensing by Exogenous Addition
Authors: Norihiro Kato, Yuriko Takayama
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Some gram-negative bacteria enable the simultaneous activation of gene expression involved in N-acylhomoserine lactone (AHL) dependent cell-to-cell communication system. Such regulatory system for the bacterial group behavior is termed as quorum sensing (QS) because a diffusible AHL signal can accumulate around the cell during the increase of the cell density and trigger activation of the sequential QS process. By blocking the QS, the expression of diverse genes related to infection, antibiotic production, and biofilm formation is inhibited. Conditioning of QS by regulation of the DNA-receptor-AHL interaction is a potential target for enhancing host defenses against pathogenicity. We focused on engineered application of transcriptional regulator SpnR produced in opportunistic human pathogen Serratia marcescens. The SpnR can interact with AHL signals at an N-terminal domain and also with a promoter region of a QS target gene at a C-terminal domain. As the initial process of the QS activation, the SpnR forms a complex with the AHL to enhance the expression of pig cluster; the SpnR normally acts as an activator for the expression of the QS-dependent gene. In this research, we attempt to artificially control QS by changing the role of SpnR. The QS-dependent prodigiosin production is expected to inhibit by externally added SpnR in the culture broth of AS-1 strain because the AHL concentration was kept below the threshold by AHL-SpnR complex formation. Maltose-binding protein (MBP)-tagged SpnR (MBP-SpnR) was overexpressed in Escherichia coli and purified using an affinity chromatography equipped with an amylose resin column. The specific interaction between AHL and MBP-SpnR was demonstrated by quartz crystal microbalance (QCM) sensor. AHL with amino end-group was coupled with COOH-terminated self-assembled monolayer prepared on a gold electrode of 27-MHz quartz crystal sensor using water-soluble carbodiimide. After the injection of MBP-SpnR into a cup-type sensor cell filled with the buffer solution, time course of resonant frequency change (ΔFs) was determined. A decrease of ΔFs clearly showed the uptake of MBP-SpnR onto the AHL-immobilized electrode. Furthermore, no binding affinity was observed after the heat-inactivation of MBP-SpnR at 80ºC. These results suggest that MBP-SpnR possesses a specific affinity for AHL. MBP-SpnR was added to the culture medium as an AHL trap to study inhibitory effects on intracellularly accumulated prodigiosin. With approximately 2 µM MBP-SpnR, the amount of prodigiosin induced was half that of the control without any additives. In conclusion, the function of SpnR could be switched by adding it to the cell culture. Exogenously added MBP-SpnR possesses high affinity for AHL derived from cells and acts as an inhibitor of AHL-mediated QS.Keywords: intracellular signaling, microbial biotechnology, quorum sensing, transcriptional regulator
Procedia PDF Downloads 2604141 Cdk1 Gates Cell Cycle-Dependent tRNA Synthesis by Regulating RNA Polymerase III Activity
Authors: Maricarmen Herrera, Pierre Chymkowitch, Joe Robertson, Jens Eriksson, Jorrit Enserink
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tRNA genes are transcribed by RNA polymerase III. During recent years, it has become clear that tDNA transcription fluctuates during the cell cycle. However, the mechanism by which the cell cycle controls the amplitude of tDNA transcription remains unknown. We found that the cyclin Clb5 recruits the cyclin dependent kinase Cdk1 to tRNA genes to sharply increase tRNA synthesis during a brief interval in the cell cycle. We show that Cdk1 promotes the interaction of TFIIIB with TFIIIC, that it stimulates the recruitment of TFIIIC to tRNA genes, that it prevents the formation of an overly stable TFIIIB-tDNA complex and that it augments the dynamics of RNA polymerase III. Furthermore, we identify Bdp1 as a novel Cdk1 substrate, and phosphorylation of Bdp1 is required for the cell cycle-dependent increase in tDNA transcription. In addition, we show that phosphorylation of the Cdk1 substrate Nup60 mediates formation of a Nup60-Nup2 complex at tRNA genes, which is also required for cell cycle-dependent tDNA transcription. Together, our findings indicate that Cdk1 activity gates tRNA synthesis by regulating the dynamics of the TFIIIB-TFIIIC-RNAPIII complex, and that it may promote the formation of a nuclear pore microenvironment conducive to efficient tDNA transcription.Keywords: Cdk1, cell cycle, RNAPIII machinery, tRNA
Procedia PDF Downloads 1764140 2D and 3D Breast Cancer Cells Behave Differently to the Applied Free Palbociclib or the Palbociclib-Loaded Nanoparticles
Authors: Maryam Parsian, Pelin Mutlu, Ufuk Gunduz
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Two-dimensional cell culture affords simplicity and low cost, but it has serious limitations; lacking cell-cell and cell-matrix interactions that are present in tissues. Cancer cells grown in 3D culture systems have distinct phenotypes of adhesion, growth, migration, invasion as well as profiles of gene and protein expression. These interactions cause the 3D-cultured cells to acquire morphological and cellular characteristics relevant to in vivo tumors. Palbociclib is a chemotherapeutic agent for the treatment of ER-positive and HER-negative metastatic breast cancer. Poly-amidoamine (PAMAM) dendrimer is a well-defined, special three-dimensional structure and has a multivalent surface and internal cavities that can play an essential role in drug delivery systems. In this study, palbociclib is loaded onto the magnetic PAMAM dendrimer. Hanging droplet method was used in order to form 3D spheroids. The possible toxic effects of both free drug and drug loaded nanoparticles were evaluated in 2D and 3D MCF-7, MD-MB-231 and SKBR-3 breast cancer cell culture models by performing MTT cell viability and Alamar Blue assays. MTT analysis was performed with six different doses from 1000 µg/ml to 25 µg/ml. Drug unloaded PAMAM dendrimer did not demonstrate significant toxicity on all breast cancer cell lines. The results showed that 3D spheroids are clearly less sensitive than 2D cell cultures to free palbociclib. Also, palbociclib loaded PAMAM dendrimers showed more toxic effect than free palbociclib in all cell lines at 2D and 3D cultures. The results suggest that the traditional cell culture method (2D) is insufficient for mimicking the actual tumor tissue. The response of the cancer cells to anticancer drugs is different in the 2D and 3D culture conditions. This study showed that breast cancer cells are more resistant to free palbociclib in 3D cultures than in 2D cultures. However, nanoparticle loaded drugs can be more cytotoxic when compared to free drug.Keywords: 2D and 3D cell culture, breast cancer, palbociclibe, PAMAM magnetic nanoparticles
Procedia PDF Downloads 1434139 The Methods of Immobilization of Laccase for Direct Transfer in an Enzymatic Fuel Cell
Authors: Afshin Farahbakhsh, Hoda Khodadadi
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In this paper, we compare five methods of biological fuel cell fabrication by combining a Shewanella oneidensis microbial anode and a laccase-modified air-breathing cathode. As a result of biofuel cell laccase with graphite nanofibers, carbon surface (PAMAN) on the pt/hpg electrode, graphite sheets MWCNT and with (PG) and (MWCNT) showed, respectively. Describes methods for creating controllable and reproducible bio-anodes and demonstrates the versatility of hybrid biological fuel cells. The laccase-based biocathodes prepared either with the crude extract or with the purified enzyme can provide electrochemically active and stable biomaterials. The laccase-based biocathodes prepared either with the crude extract or with the purified enzyme can provide electrochemically active and stable biomaterials. When the device was fed with transdermal extracts, containing only 30μM of glucose, the average peak power was proportionally lower (0.004mW). The result of biofuel cell with graphite nanofibers showed the enzymatic fuel cell reaches 0.5 V at open circuit voltage with both, ethanol and methanol and the maximum current density observed for E2electrode was 228.94mAcm.Keywords: enzymatic electrode, fuel cell, immobilization, laccase
Procedia PDF Downloads 2554138 Anti-Proliferative Effect of Chanterelle (Cantharellus) Mushroom Extracts on Glioblastoma Multiforme Cell Line U87MG
Authors: Justyna Moskwa, Patryk Nowakowski, Sylwia K. Naliwajko, Renata Markiewicz-Zukowska, Krystyna Gromkowska-Kepka, Anna Puscion-Jakubik, Konrad Mielcarek, Maria H. Borawska
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For centuries, mushrooms have been used in folk medicine; however, knowledge of the composition and properties of fungi comes from the last twenty years. Mushrooms show antibacterial, antioxidant, antitumor and immune-stimulating properties; however, there is a lack of reports, on anticancer treatment of brain gliomas. The aim of this study was to examine influence of Chanterelle mushroom (Cantharellus Adans. ex Fr.) ethanolic (CHE) and water (CHW) extracts, on glioblastoma multiforme cell line (U87MG). Anti-proliferative activity of CHE and CHW in concentration (50-1000 µg/mL) was determined by a cytotoxicity test and DNA binding by [³H]-thymidine incorporation after 24, 48 and 72h of incubation with U87MG glioblastoma cell line. The statistical analysis was performed using Statistica v. 13.0 software. Significant differences were assumed for p < 0.05. We examined that CHE extracts in all the tested concentrations (50, 100, 250, 500, 1000 µg/mL) after all hours of incubation significantly decreased cell viability (p < 0.05) on U87MG cell line, which was confirmed by the significant (p < 0.05) reduction of DNA synthesis. Our results suggest that only CHE extract a cytotoxic and anti-proliferation activities on U87MG cell line.Keywords: anticancer, food, glioblastoma, mushroom
Procedia PDF Downloads 1534137 Environmental Pb-Free Cu Front Electrode for Si-Base Solar Cell Application
Authors: Wen-Hsi Lee, C.G. Kao
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In this study, Cu paste was prepared and printed with narrow line screen printing process on polycrystalline Si solar cell which has already finished the back Al printing and deposition of double anti-reflection coatings (DARCs). Then, two-step firing process was applied to sinter the front electrode and obtain the ohmic contact between front electrode and solar cell. The first step was in air atmosphere. In this process, PbO-based glass frit etched the DARCs and Ag recrystallized at the surface of Si, constructing the preliminary contact. The second step was in reducing atmosphere. In this process, CuO reduced to Cu and sintered. Besides, Ag nanoparticles recrystallized in the glass layer at interface due to the interactions between H2, Ag and PbO-based glass frit and the volatility of Pb, constructing the ohmic contact between electrode and solar cell. By experiment and analysis, reaction mechanism in each stage was surmised, and it was also proven that ohmic contact and good sheet resistance for front electrode could both be obtained by applying newly-invented paste and process.Keywords: front electrode, solar cell, ohmic contact, screen printing, paste
Procedia PDF Downloads 3284136 Analysis of Cell Cycle Status in Radiation Non-Targeted Hepatoma Cells Using Flow Cytometry: Evidence of Dose Dependent Response
Authors: Sharmi Mukherjee, Anindita Chakraborty
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Cellular irradiation incites complex responses including arrest of cell cycle progression. This article accentuates the effects of radiation on cell cycle status of radiation non-targeted cells. Human Hepatoma HepG2 cells were exposed to increasing doses of γ radiations (1, 2, 4, 6 Gy) and their cell culture media was transferred to non-targeted HepG2 cells cultured in other Petri plates. These radiation non-targeted cells cultured in the ICCM (Irradiated cell conditioned media) were the bystander cells on which cell cycle analysis was performed using flow cytometry. An apparent decrease in the distribution of bystander cells at G0/G1 phase was observed with increased radiation doses upto 4 Gy representing a linear relationship. This was accompanied by a gradual increase in cellular distribution at G2/M phase. Interestingly the number of cells in G2/M phase at 1 and 2 Gy irradiation was not significantly different from each other. However, the percentage of G2 phase cells at 4 and 6 Gy doses were significantly higher than 2 Gy dose indicating the IC50 dose to be between 2 and 4 Gy. Cell cycle arrest is an indirect indicator of genotoxic damage in cells. In this study, bystander stress signals through the cell culture media of irradiated cells disseminated the radiation induced DNA damages in the non-targeted cells which resulted in arrest of the cell cycle progression at G2/M phase checkpoint. This implies that actual radiation biological effects represent a penumbra with effects encompassing a larger area than the actual beam. This article highlights the existence of genotoxic damages as bystander effects of γ rays in human Hepatoma cells by cell cycle analysis and opens up avenues for appraisal of bystander stress communications between tumor cells. Contemplation of underlying signaling mechanisms can be manipulated to maximize damaging effects of radiation with minimum dose and thus has therapeutic applications.Keywords: bystander effect, cell cycle, genotoxic damage, hepatoma
Procedia PDF Downloads 1784135 First Investigation on CZTS Electron affinity and Thickness Optimization using SILVACO-Atlas 2D Simulation
Authors: Zeineb Seboui, Samar Dabbabi
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In this paper, we study the performance of Cu₂ZnSnS₄ (CZTS) based solar cell. In our knowledge, it is for the first time that the FTO/ZnO:Co/CZTS structure is simulated using the SILVACO-Atlas 2D simulation. Cu₂ZnSnS₄ (CZTS), ZnO:Co and FTO (SnO₂:F) layers have been deposited on glass substrates by the spray pyrolysis technique. The extracted physical properties, such as thickness and optical parameters of CZTS layer, are considered to create a new input data of CZTS based solar cell. The optimization of CZTS electron affinity and thickness is performed to have the best FTO/ZnO: Co/CZTS efficiency. The use of CZTS absorber layer with 3.99 eV electron affinity and 3.2 µm in thickness leads to the higher efficiency of 16.86 %, which is very important in the development of new technologies and new solar cell devices.Keywords: CZTS solar cell, characterization, electron affinity, thickness, SILVACO-atlas 2D simulation
Procedia PDF Downloads 714134 Frequency of Alloimmunization in Sickle Cell Disease Patients in Africa: A Systematic Review with Meta-analysis
Authors: Theresa Ukamaka Nwagha, Angela Ogechukwu Ugwu, Martins Nweke
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Background and Objectives: Blood transfusion is an effective and proven treatment for some severe complications of sickle cell disease. Recurrent transfusions have put patients with sickle cell disease at risk of developing antibodies against the various antigens they were exposed to. This study aims to investigate the frequency of red blood cell alloimmunization in patients with sickle disease in Africa. Materials and Methods: This is a systematic review of peer-reviewed literature published in English. The review was conducted consistent with the Preferred Reporting Items for Systematic Reviews and Meta-Analyses checklist. Data sources for the review include MEDLINE, PubMed, CINAHL, and Academic Search Complete. Included in this review are articles that reported the frequency/prevalence of red blood cell alloimmunization in sickle cell disease patients in Africa. Eligible studies were subjected to independent full-text screening and data extraction. Risk of bias assessment was conducted with the aid of the mixed method appraisal tool. We employed a random-effects model of meta-analysis to estimate the pooled prevalence. We computed Cochrane’s Q statistics and I2 and prediction interval to quantify heterogeneity in effect size. Results: The prevalence estimates range from 2.6% to 29%. Pooled prevalence was estimated to be 10.4% (CI 7.7.–13.8); PI = 3.0 – 34.0%), with significant heterogeneity (I2 = 84.62; PI = 2.0-32.0%) and publication bias (Egger’s t-test = 1.744, p = 0.0965). Conclusion: The frequency of red cell alloantibody varies considerably in Africa. The alloantibodies appeared frequent in this order: the Rhesus, Kell, Lewis, Duffy, MNS, and LutheranKeywords: frequency, red blood cell, alloimmunization, sickle cell disease, Africa
Procedia PDF Downloads 924133 Induction of HIV-1 Resistance: The New Approaches Based on Gene Modification and Stem Cell Engineering
Authors: Alieh Farshbaf
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Introduction: Current anti-retroviral drugs have some restrictions for treatment of HIV-1 infection. The efficacy of retroviral drugs is not same in different infected patients and the virus rebound from latent reservoirs after stopping them. Recently, the engineering of stem cells and gene therapy provide new approaches to eliminate some drug problems by induction of resistance to HIV-1. Literature review: Up to now, AIDS-restriction genes (ARGs) were suitable candidate for gene and cell therapies, such as cc-chemokine receptor-5 (CCR5). In this manner, CCR5 provide effective cure in Berlin and Boston patients by inducing of HIV-1 resistance with allogeneic stem cell transplantation. It is showed that Zinc Finger Nuclease (ZFN) could induce HIV-1 resistance in stem cells of infected patients by homologous recombination or non-end joining mechanism and eliminate virus loading after returning the modified cells. Then, gene modification by HIV restriction factors, as TRIM5, introduced another gene candidate for HIV by interfering in infection process. These gene modifications/editing provided by stem cell futures that improve treatment in refractory disease such as HIV-1. Conclusion: Although stem cell transplantation has some complications, but in compare to retro-viral drugs demonstrated effective cure by elimination of virus loading. On the other hand, gene therapy is cost-effective for an infected patient than retroviral drugs payment in a person life-long. The results of umbilical cord blood stem cell transplantation showed that gene and cell therapy will be applied easier than previous treatment of AIDS with high efficacy.Keywords: stem cell, AIDS, gene modification, cell engineering
Procedia PDF Downloads 2974132 MTT Assay-Guided Isolation of a Cytotoxic Lead from Hedyotis umbellata and Its Mechanism of Action against Non-Small Cell Lung Cancer A549 Cells
Authors: Kirti Hira, A. Sajeli Begum, S. Mahibalan, Poorna Chandra Rao
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Introduction: Cancer is one of the leading causes of death worldwide. Although existing therapy effectively kills cancer cells, they do affect normal growing cells leading to many undesirable side effects. Hence there is need to develop effective as well as safe drug molecules to combat cancer, which is possible through phyto-research. The currently available plant-derived blockbuster drugs are the example for this. In view of this, an investigation was done to identify cytotoxic lead molecules from Hedyotis umbellata (Family Rubiaceae), a widely distributed weed in India. Materials and Methods: The methanolic extract of the whole plant of H. umbellata (MHU), prepared through Soxhlet extraction method was further fractionated with diethyl ether and n-butanol, successively. MHU, ether fraction (EMHU) and butanol fraction (BMHU) were lyophilized and were tested for the cytotoxic effect using 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay against non-small cell lung cancer (NSCLC) A549 cell lines. The potentially active EMHU was subjected to chromatographic purification using normal-phase silica columns, in order to isolate the responsible bioactive compounds. The isolated pure compounds were tested for their cytotoxic effect by MTT assay against A549 cells. Compound-3, which was found to be most active, was characterized using IR, 1H- and 13C-NMR and MS analysis. The study was further extended to decipher the mechanism of action of cytotoxicity of compound-3 against A549 cells through various in vitro cellular models. Cell cycle analysis was done using flow cytometry following PI (Propidium Iodide) staining. Protein analysis was done using Western blot technique. Results: Among MHU, EMHU, and BMHU, the non-polar fraction EMHU demonstrated a significant dose-dependent cytotoxic effect with IC50 of 67.7μg/ml. Chromatography of EMHU yielded seven compounds. MTT assay of isolated compounds explored compound-3 as potentially active one, which inhibited the growth of A549 cells with IC50value of 14.2μM. Further, compound-3 was identified as cedrelopsin, a coumarin derivative having molecular weight of 260. Results of in vitro mechanistic studies explained that cedrelopsin induced cell cycle arrest at G2/M phase and down-regulated the expression of G2/M regulatory proteins such as cyclin B1, cdc2, and cdc25C, dose dependently. This is the first report that explores the cytotoxic mechanism of cedrelopsin. Conclusion: Thus a potential small lead molecule, cedrelopsin isolated from H. umbellata, showing antiproliferative effect mediated by G2/M arrest in A549 cells was discovered. The effect of cedrelopsin against other cancer cell lines followed by in vivo studies can be performed in future to develop a new drug candidate.Keywords: A549, cedrelopsin, G2/M phase, Hedyotis umbellata
Procedia PDF Downloads 1704131 Study on the Effect of Bolt Locking Method on the Deformation of Bipolar Plate in PEMFC
Authors: Tao Chen, ShiHua Liu, JiWei Zhang
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Assembly of the proton exchange membrane fuel cells (PEMFC) has a very important influence on its performance and efficiency. The various components of PEMFC stack are usually locked and fixed by bolts. Locking bolt will cause the deformation of the bipolar plate and the other components, which will affect directly the deformation degree of the integral parts of the PEMFC as well as the performance of PEMFC. This paper focuses on the object of three-cell stack of PEMFC. Finite element simulation is used to investigate the deformation of bipolar plate caused by quantity and layout of bolts, bolt locking pressure, and bolt locking sequence, etc. Finally, we made a conclusion that the optimal combination packaging scheme was adopted to assemble the fuel cell stack. The scheme was in use of 3.8 MPa locking pressure imposed on the fuel cell stack, type Ⅱ of four locking bolts and longitudinal locking method. The scheme was obtained by comparatively analyzing the overall displacement contour of PEMFC stack, absolute displacement curve of bipolar plate along the given three paths in the Z direction and the polarization curve of fuel cell. The research results are helpful for the fuel cell stack assembly.Keywords: bipolar plate, deformation, finite element simulation, fuel cell, locking bolt
Procedia PDF Downloads 4094130 Studying the Antiapoptotic Activity of Β Cells from Cord Blood Based Mesenchymal Stem Cells as an Approach to Treat Diabetes Mellitus
Authors: Parcha Sreenivasa Rao, P. Lakshmi
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Diabetes Mellitus is metabolic disorder, characterized by high glucose levels in the blood due to one of the reason i.e., the death of β cells. The lack of β cells leads to the reduced insulin levels. The β cell death generally occurs due to apoptosis induced by the several cytokines. IL-1β, IFN- ϒ and TNF –α cytokines that are generally cause apoptosis to the β cell. The nutrient based apoptosis is generally seen with high glucose and free fatty acids. It is also noted that the β cell death triggered by Fas ligand and its receptor Fas at the surface of the activated CD8+ T- lymphocytes. Reports also reveal that the β cell apoptosis is under control of the transcription factors NF-kB and STAT- 1. The arresting or opposing of the β cell apoptosis can be overcome by the different growth factors like GLP-1, growth hormone, prolactin, VEGF, Dipeptidyl peptidase-4, Vildagliptin, suberoylanilidehydroxamic acid, trichistatin-A, XIAP, Bcl-2, FGF-21. Present investigation explains antiapoptotic property of the β cells derived from the mesenchymal stem cells of umbilical cord.Keywords: stem cells, umblical cord, diabetes, apoptosis
Procedia PDF Downloads 3744129 RhoA Regulates E-Cadherin Intercellular Junctions in Oral Squamous Carcinoma Cells
Authors: Ga-Young Lee, Hyun-Man Kim
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The modulation of the cell-cell junction is critical in epithelial-mesenchymal transition during tumorigenesis. As RhoA activity is known to be up-regulated to dissociate cell-cell junction by contracting acto-myosin complex in various cancer cells, the present study investigated if RhoA activity was also associated with the disruption of the cell-cell junction of oral cancer cells. We studied SCC-25 cells which are established from oral squamous cell carcinoma if their E-cadherin junction (ECJ) was under control of RhoA. Interestingly, development of ECJ of SCC-25 cells depended on the amount of fibronectin (FN) coated on the culture dishes. Seeded cells promptly aggregated to develop ECJ on the substrates coated with a low amount of FN, whereas they were retarded in the development of ECJ on the substrates coated with a high amount of FN. However, it was an unexpected finding that total RhoA activity was lower in the dissociated cells on the substrates of high FN than in the aggregated cells on the substrates of low FN. Treating the dissociated cells on the substrates of high FN with LPA, a RhoA activator, promoted the development to ECJ. In contrast, treating the aggregated cells on the substrates of low FN with Clostridium botulinum C3, a toxin decreasing RhoA activity, dissociated cells concomitant with the disruption of ECJ. Genetical knockdown of RhoA expression by transfecting RhoA siRNA also down-regulated the development of ECJ in SCC-25 cells. Furthermore, PMA, an activator of protein kinase C (PKC), down-regulated the development of ECJ junction of SCC-25 cells on the substrates coated with low FN. In contrast, GO6976, a PKC inhibitor, up-regulated the development of ECJ of SCC-25 cells with the activation of RhoA on the substrates coated with high FN. In conclusion, in the present study, we demonstrated unexpected results that the activation of RhoA promotes the development of ECJ, whereas the inhibition of RhoA retards the development of ECJ in SCC-25 cells.Keywords: E-cadherin junction, oral squamous cell carcinoma, PKC, RhoA, SCC-25
Procedia PDF Downloads 3254128 Biophysically Motivated Phylogenies
Authors: Catherine Felce, Lior Pachter
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Current methods for building phylogenetic trees from gene expression data consider mean expression levels. With single-cell technologies, we can leverage more information about cell dynamics by considering the entire distribution of gene expression across cells. Using biophysical modeling, we propose a method for constructing phylogenetic trees from scRNA-seq data, building on Felsenstein's method of continuous characters. This method can highlight genes whose level of expression may be unchanged between species, but whose rates of transcription/decay may have evolved over time.Keywords: phylogenetics, single-cell, biophysical modeling, transcription
Procedia PDF Downloads 374127 Conformal Coating Technology Applicable to Cell Therapeutics Using Click-Reactive Biocompatible Polymers
Authors: Venkat Garigapati
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Cell-based therapies are limited due to underlying host immune system activity. Microencapsulation of living cells to overcome this issue has some serious drawbacks, such as limitations of nutrient and oxygen diffusion, which pose a threat to the function and longevity of cells. The conformal coating could overcome the issues which are generally involved in traditional microencapsulation. Some of the theoretical advantages of conformal coating include superior nutrient and oxygen supply to cells, prolonged lifespan, improved drug-secreting cell functionality and an opportunity to load high cell doses in small volumes. Despite several advantages to the conformal coating, there are no suitable methods available to apply to living cells. The ultra-thin conformal coating was achieved utilizing click-reactive methacryloyloxyethyl phosphorylcholine (MPC) polymers, which are capable of specifically reacting one polymer to another at neutral pH in the aqueous isotonic system at the desired temperature suitable for living cells without the need of deleterious initiators. ARPE-19 (Adult Retinal Pigment Epithelial cell line-19) cell-spheroids and rat pancreatic islets were used in the formulation studies. The in vitro studies of coated ARPE-19 cell-spheroids and rat islets indicate that the coat was intact; cells were viable and functioning. The in vitro study results revealed that the conformal coating technology seems promising and in vivo studies are being planned.Keywords: cells, hydrogel, conformal coating, microencapsulation, insulin
Procedia PDF Downloads 874126 Sensitivity Studies for a Pin Homojunction a-Si:H Solar Cell
Authors: Leila Ayat, Afak Meftah
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Amorphous-silicon alloys have great promise as low cost solar cell materials. They have excellent photo-conductivity and high optical absorption to sunlight. Now PIN a-Si:H based solar cells are widely used in power generation modules. However, to improve the performance of these cells further, a better fundamental under-standing of the factors limiting cell performance in the homo junction PIN structure is necessary. In this paper we discuss the sensitivity of light J-V characteristics to various device and material parameters in PIN homo junction solar cells. This work is a numerical simulation of the output parameters of a PIN a-Si:H solar cell under AM1.5 spectrum. These parameters are the short circuit current (Jsc), the open circuit voltage (Voc), the fill factor (FF), the conversion efficiency. The simulation was performed with SCAPS-1D software version 3.3 developed at ELIS in Belgium by Marc Burgelman et al. The obtained results are in agreement with experiment. In addition, the effect of the thickness, doping density, capture cross sections of the gap states and the band microscopic mobilities on the output parameters of the cell are also presented.Keywords: amorphous silicon p-i-n junctions, thin film, solar cells, sensitivity
Procedia PDF Downloads 5164125 Mitigating the Aggregation of Human Islet Amyloid Polypeptide with Nanomaterials
Authors: Ava Faridi, Pouya Faridi, Aleksandr Kakinen, Ibrahim Javed, Thomas P. Davis, Pu Chun Ke
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Human islet amyloid polypeptide (IAPP) is a hormone associated with glycemic control and type 2 diabetes. Biophysically, the chirality of IAPP fibrils has been little explored with respect to the aggregation and toxicity of the peptide. Biochemically, it remains unclear as for how protein expression in pancreatic beta cells may be altered by cell exposure to the peptide, and how such changes may be mitigated by nanoparticle inhibitors for IAPP aggregation. In this study, we first demonstrated the elimination of the IAPP nucleation phase and shortening of its elongation phase by silica nanoribbons. This accelerated IAPP fibrillization translated to reduced toxicity, especially for the right-handed silica nanoribbons, as revealed by cell viability, helium ion microscopy, as well as zebrafish embryo survival, developmental and behavioral assays. We then examined the proteomes of βTC6 pancreatic beta cells exposed to the three main aggregation states of monomeric, oligomeric and amyloid fibrillar IAPP, and compared that with cellular protein expression modulated by graphene quantum dots (GQDs). A total of 29 proteins were significantly regulated by different forms of IAPP, and the majority of these proteins were nucleotide-binding proteins. A regulatory capacity of GQDs against aberrant protein expression was confirmed. These studies have demonstrated the great potential of employing nanomaterials targeting the mesoscopic enantioselectivity and protein expression dysregulation in pancreatic beta cells.Keywords: graphene quantum dots, IAPP, silica nanoribbons, protein expression, toxicity
Procedia PDF Downloads 1364124 Camptothecin Promotes ROS-Mediated G2/M Phase Cell Cycle Arrest, Resulting from Autophagy-Mediated Cytoprotection
Authors: Rajapaksha Gedara Prasad Tharanga Jayasooriya, Matharage Gayani Dilshara, Yung Hyun Choi, Gi-Young Kim
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Camptothecin (CPT) is a quinolone alkaloid which inhibits DNA topoisomerase I that induces cytotoxicity in a variety of cancer cell lines. We previously showed that CPT effectively inhibited invasion of prostate cancer cells and also combined treatment with subtoxic doses of CPT and TNF-related apoptosis-inducing ligand (TRAIL) potentially enhanced apoptosis in a caspase-dependent manner in hepatoma cancer cells. Here, we found that treatment with CPT caused an irreversible cell cycle arrest in the G2/M phase. CPT-induced cell cycle arrest was associated with a decrease in protein levels of cell division cycle 25C (Cdc25C) and increased the level of cyclin B and p21. The CPT-induced decrease in Cdc25C was blocked in the presence of proteasome inhibitor MG132, thus reversed the cell cycle arrest. In addition to that treatment of CPT-increased phosphorylation of Cdc25C was the resulted of activation of checkpoint kinase 2 (Chk2), which was associated with phosphorylation of ataxia telangiectasia-mutated. Interestingly CPT induced G2/M phase of the cell cycle arrest is reactive oxygen species (ROS) dependent where ROS inhibitors NAC and GSH reversed the CPT-induced cell cycle arrest. These results further confirm by using transient knockdown of nuclear factor-erythroid 2-related factor 2 (Nrf2) since it regulates the production of ROS. Our data reveal that treatment of siNrf2 increased the ROS level as well as further increased the CPT induce G2/M phase cell cycle arrest. Our data also indicate CPT-enhanced cell cycle arrest through the extracellular signal-regulated kinase (ERK) and the c-Jun N-terminal kinase (JNK) pathway. Inhibitors of ERK and JNK more decreased the Cdc25C expression and protein expression of p21 and cyclin B. These findings indicate that Chk2-mediated phosphorylation of Cdc25C plays a major role in G2/M arrest by CPT.Keywords: camptothecin, cell cycle, checkpoint kinase 2, nuclear factor-erythroid 2-related factor 2, reactive oxygen species
Procedia PDF Downloads 4284123 Following the Modulation of Transcriptional Activity of Genes by Chromatin Modifications during the Cell Cycle in Living Cells
Authors: Sharon Yunger, Liat Altman, Yuval Garini, Yaron Shav-Tal
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Understanding the dynamics of transcription in living cells has improved since the development of quantitative fluorescence-based imaging techniques. We established a method for following transcription from a single copy gene in living cells. A gene tagged with MS2 repeats, used for mRNA tagging, in its 3' UTR was integrated into a single genomic locus. The actively transcribing gene was detected and analyzed by fluorescence in situ hybridization (FISH) and live-cell imaging. Several cell clones were created that differed in the promoter regulating the gene. Thus, comparative analysis could be obtained without the risk of different position effects at each integration site. Cells in S/G2 phases could be detected exhibiting two adjacent transcription sites on sister chromatids. A sharp reduction in the transcription levels was observed as cells progressed along the cell cycle. We hypothesized that a change in chromatin structure acts as a general mechanism during the cell cycle leading to down-regulation in the activity of some genes. We addressed this question by treating the cells with chromatin decondensing agents. Quantifying and imaging the treated cells suggests that chromatin structure plays a role both in regulating transcriptional levels along the cell cycle, as well as in limiting an active gene from reaching its maximum transcription potential at any given time. These results contribute to understanding the role of chromatin as a regulator of gene expression.Keywords: cell cycle, living cells, nucleus, transcription
Procedia PDF Downloads 2934122 Comparison of Extracellular miRNA from Different Lymphocyte Cell Lines and Isolation Methods
Authors: Christelle E. Chua, Alicia L. Ho
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The development of a panel of differential gene expression signatures has been of interest in the field of biomarker discovery for radiation exposure. In the absence of the availability of exposed human subjects, lymphocyte cell lines have often been used as a surrogate to human whole blood, when performing ex vivo irradiation studies. The extent of variation between different lymphocyte cell lines is currently unclear, especially with regard to the expression of extracellular miRNA. This study compares the expression profile of extracellular miRNA isolated from different lymphocyte cell lines. It also compares the profile of miRNA obtained when different exosome isolation kits are used. Lymphocyte cell lines were created using lymphocytes isolated from healthy adult males of similar racial descent (Chinese American and Chinese Singaporean) and immortalised with Epstein-Barr virus. The cell lines were cultured in exosome-free cell culture media for 72h and the cell culture supernatant was removed for exosome isolation. Two exosome isolation kits were used. Total exosome isolation reagent (TEIR, ThermoFisher) is a polyethylene glycol (PEG)-based exosome precipitation kit, while ExoSpin (ES, Cell Guidance Systems) is a PEG-based exosome precipitation kit that includes an additional size exclusion chromatography step. miRNA from the isolated exosomes were isolated using miRNEASY minikit (Qiagen) and analysed using nCounter miRNA assay (Nanostring). Principal component analysis (PCA) results suggested that the overall extracellular miRNA expression profile differed between the lymphocyte cell line originating from the Chinese American donor and the cell line originating from the Chinese Singaporean donor. As the gender, age and racial origins of both donors are similar, this may suggest that there are other genetic or epigenetic differences that account for the variation in extracellular miRNA gene expression in lymphocyte cell lines. However, statistical analysis showed that only 3 miRNA genes had a fold difference > 2 at p < 0.05, suggesting that the differences may not be of that great a significance as to impact overall conclusions drawn from different cell lines. Subsequent analysis using cell lines from other donors will give further insight into the reproducibility of results when difference cell lines are used. PCA results also suggested that the method of exosome isolation impacted the expression profile. 107 miRNA had a fold difference > 2 at p < 0.05. This suggests that the inclusion of an additional size exclusion chromatography step altered the subset of the extracellular vesicles that were isolated. In conclusion, these results suggest that extracellular miRNA can be isolated and analysed from exosomes derived from lymphocyte cell lines. However, care must be taken in the choice of cell line and method of exosome isolation used.Keywords: biomarker, extracellular miRNA, isolation methods, lymphocyte cell line
Procedia PDF Downloads 1914121 Spectral Clustering for Manufacturing Cell Formation
Authors: Yessica Nataliani, Miin-Shen Yang
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Cell formation (CF) is an important step in group technology. It is used in designing cellular manufacturing systems using similarities between parts in relation to machines so that it can identify part families and machine groups. There are many CF methods in the literature, but there is less spectral clustering used in CF. In this paper, we propose a spectral clustering algorithm for machine-part CF. Some experimental examples are used to illustrate its efficiency. Overall, the spectral clustering algorithm can be used in CF with a wide variety of machine/part matrices.Keywords: group technology, cell formation, spectral clustering, grouping efficiency
Procedia PDF Downloads 395