Search results for: cell culture passage
6888 Deciphering Tumor Stroma Interactions in Retinoblastoma
Authors: Rajeswari Raguraman, Sowmya Parameswaran, Krishnakumar Subramanian, Jagat Kanwar, Rupinder Kanwar
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Background: Tumor microenvironment has been implicated in several cancers to regulate cell growth, invasion and metastasis culminating in outcome of therapy. Tumor stroma consists of multiple cell types that are in constant cross-talk with the tumor cells to favour a pro-tumorigenic environment. Not much is known about the existence of tumor microenvironment in the pediatric intraocular malignancy, Retinoblastoma (RB). In the present study, we aim to understand the multiple stromal cellular subtypes and tumor stromal interactions expressed in RB tumors. Materials and Methods: Immunohistochemistry for stromal cell markers CD31, CD68, alpha-smooth muscle (α-SMA), vimentin and glial fibrillary acidic protein (GFAP) was performed on formalin fixed paraffin embedded tissues sections of RB (n=12). The differential expression of stromal target molecules; fibroblast activation protein (FAP), tenascin-C (TNC), osteopontin (SPP1), bone marrow stromal antigen 2 (BST2), stromal derived factor 2 and 4 (SDF2 and SDF4) in primary RB tumors (n=20) and normal retina (n=5) was studied by quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) and Western blotting. The differential expression was correlated with the histopathological features of RB. The interaction between RB cell lines (Weri-Rb-1, NCC-RbC-51) and Bone marrow stromal cells (BMSC) was also studied using direct co-culture and indirect co-culture methods. The functional effect of the co-culture methods on the RB cells was evaluated by invasion and proliferation assays. Global gene expression was studied by using Affymetrix 3’ IVT microarray. Pathway prediction was performed using KEGG and the key molecules were validated using qRT-PCR. Results: The immunohistochemistry revealed the presence of several stromal cell types such as endothelial cells (CD31+;Vim+/-); macrophages (CD68+;Vim+/-); Fibroblasts (Vim+; CD31-;CD68- );myofibroblasts (α-SMA+/ Vim+) and invading retinal astrocytes/ differentiated retinal glia (GFAP+; Vim+). A characteristic distribution of these stromal cell types was observed in the tumor microenvironment, with endothelial cells predominantly seen in blood vessels and macrophages near actively proliferating tumor or necrotic areas. Retinal astrocytes and glia were predominant near the optic nerve regions in invasive tumors with sparse distribution in tumor foci. Fibroblasts were widely distributed with rare evidence of myofibroblasts in the tumor. Both gene and protein expression revealed statistically significant (P<0.05) up-regulation of FAP, TNC and BST2 in primary RB tumors compared to the normal retina. Co-culture of BMSC with RB cells promoted invasion and proliferation of RB cells in direct and indirect contact methods respectively. Direct co-culture of RB cell lines with BMSC resulted in gene expression changes in ECM-receptor interaction, focal adhesion, IL-8 and TGF-β signaling pathways associated with cancer. In contrast, various metabolic pathways such a glucose, fructose and amino acid metabolism were significantly altered under the indirect co-culture condition. Conclusion: The study suggests that the close interaction between RB cells and the stroma might be involved in RB tumor invasion and progression which is likely to be mediated by ECM-receptor interactions and secretory factors. Targeting the tumor stroma would be an attractive option for redesigning treatment strategies for RB.Keywords: gene expression profiles, retinoblastoma, stromal cells, tumor microenvironment
Procedia PDF Downloads 3806887 Synthesis and Application of Oligosaccharides Representing Plant Cell Wall Polysaccharides
Authors: Mads H. Clausen
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Plant cell walls are structurally complex and contain a larger number of diverse carbohydrate polymers. These plant fibers are a highly valuable bio-resource and the focus of food, energy and health research. We are interested in studying the interplay of plant cell wall carbohydrates with proteins such as enzymes, cell surface lectins and antibodies. However, detailed molecular level investigations of such interactions are hampered by the heterogeneity and diversity of the polymers of interest. To circumvent this, we target well-defined oligosaccharides with representative structures that can be used for characterizing protein-carbohydrate binding. The presentation will highlight chemical syntheses of plant cell wall oligosaccharides from our group and provide examples from studies of their interactions with proteins.Keywords: oligosaccharides, carbohydrate chemistry, plant cell walls, carbohydrate-acting enzymes
Procedia PDF Downloads 3016886 The EAO2 in Essouabaa, Tebessa, Algeria: An Example of Facies to Organic Matter
Authors: Sihem Salmi Laouar, Khoudair Chabane, Rabah Laouar, Adrian J. Boyce et Anthony E. Fallick
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The solid mass of Essouabaa belongs paléogéography to the field téthysian and belonged to the area of the Mounts of Mellègue. This area was not saved by the oceanic-2 event anoxic (EAO-2) which was announced, over one short period, around the limit cénomanian-turonian. In the solid mass of Essouabba, the dominant sediments, pertaining to this period, are generally fine, dark, laminated and sometimes rolled deposits. They contain a rather rich planktonic microfaune, pyrite, and grains of phosphate, thus translating an environment rather deep and reducing rather deep and reducing. For targeting well the passage Cénomanian-Turonian (C-T) in the solid mass of Essouabaa, of the studies lithological and biostratigraphic were combined with the data of the isotopic analyses carbon and oxygen like with the contents of CaCO3. The got results indicate that this passage is marked by a biological event translated by the appearance of the "filaments" like by a positive excursion of the δ13C and δ18O. The cénomanian-turonian passage in the solid mass of Essouabaa represents a good example where during the oceanic event anoxic a facies with organic matter with contents of COT which can reach 1.36%. C E massive presents biostratigraphic and isotopic similarities with those obtained as well in the areas bordering (ex: Tunisia and Morocco) that throughout the world.Keywords: limit cénomanian-turonian (C-T), COT, filaments, event anoxic 2 (EAO-2), stable isotopes, mounts of Mellègue, Algeria
Procedia PDF Downloads 5096885 Isolation of the Leptospira spp. from the Rice Farming Lands in the North of Iran by EMJH Media
Authors: S. Rostampour Yasouri, M. Ghane
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Leptospirosis is one the most important common diseases between human and live stock occurred by different species of Leptospira. This disease has been construed as the native in the northern provinces of Iran and risk of the infection with pathogenic is high. One hundred fifteen samples of water (67), soil (36) and feces of rodents (12) were collected from the rice fields of the suburbs of Tonekabon Township situated in northern part of Iran in 2012. The samples, after passage from membranous filters, were cultured in the liquid and solid EMJH medium and incubated at 30°C for 1 month. Leptospira spp. were isolated using culture technique, and the plates were studied from viewpoint of colony formation, microscopic observations and then identified by phenotyping tests. Finally, the identification of Leptospira genus was verified by PCR technique and 16S rRNA gene sequencing. Of 115 samples totally, 55 samples (47.82%) became positive by use of the culture technique which the positive cases included 47 water samples (70.14%) and 8 soil samples (22.22%), while the isolation was not accomplished from the sample of the rodents feces. Overall, according to these data, Leptospira spp. exists with high frequency in North Iran. Hence, based on foregoing evidence environments in the north of Iran are vehicles of Leptospira spp.Keywords: EMJH Medium, Leptospira, Northern of Iran, rice fields
Procedia PDF Downloads 1756884 Structural Evaluation of Cell-Filled Pavement
Authors: Subrat Roy
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This paper describes the findings of a study carried out for evaluating the performance of cell-filled pavement for low volume roads. Details of laboratory investigations and the methodology adopted for construction of cell-filled pavement are presented. The aim of this study is to evaluate the structural behaviour of cement concrete filled cell pavement laid over three different types of subbases (water bound macadam, soil-cement and moorum). A formwork of cells of a thin plastic sheet was used to construct the cell-filled pavements to form flexible, interlocked block pavements. Surface deflections were measured using falling weight deflectometer and benkelman beam methods. Resilient moduli of pavement layers were estimated from the measured deflections. A comparison of deflections obtained from both the methodology is also presented.Keywords: cell-filled pavement, WBM, FWD, Moorum
Procedia PDF Downloads 2936883 Physiological Normoxia and Cellular Adhesion of Diffuse Large B-Cell Lymphoma Primary Cells: Real-Time PCR and Immunohistochemistry Study
Authors: Kamila Duś-Szachniewicz, Kinga M. Walaszek, Paweł Skiba, Paweł Kołodziej, Piotr Ziółkowski
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Cell adhesion is of fundamental importance in the cell communication, signaling, and motility, and its dysfunction occurs prevalently during cancer progression. The knowledge of the molecular and cellular processes involved in abnormalities in cancer cells adhesion has greatly increased, and it has been focused mainly on cellular adhesion molecules (CAMs) and tumor microenvironment. Unfortunately, most of the data regarding CAMs expression relates to study on cells maintained in standard oxygen condition of 21%, while the emerging evidence suggests that culturing cells in ambient air is far from physiological. In fact, oxygen in human tissues ranges from 1 to 11%. The aim of this study was to compare the effects of physiological lymph node normoxia (5% O2), and hyperoxia (21% O2) on the expression of cellular adhesion molecules of primary diffuse large B-cell lymphoma cells (DLBCL) isolated from 10 lymphoma patients. Quantitative RT-PCR and immunohistochemistry were used to confirm the differential expression of several CAMs, including ICAM, CD83, CD81, CD44, depending on the level of oxygen. Our findings also suggest that DLBCL cells maintained at ambient O2 (21%) exhibit reduced growth rate and migration ability compared to the cells growing in normoxia conditions. Taking into account all the observations, we emphasize the need to identify the optimal human cell culture conditions mimicking the physiological aspects of tumor growth and differentiation.Keywords: adhesion molecules, diffuse large B-cell lymphoma, physiological normoxia, quantitative RT-PCR
Procedia PDF Downloads 2716882 Parental Monitoring of Learners’ Cell Phone Use in the Eastern Cape, South Africa
Authors: Melikhaya Skhephe, Robert Mawuli Kwasi Boadzo, Zanoxolo Berington Gobingca
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This research study sought to examine parental monitoring of learners’ cell phone use in the Eastern Cape, South Africa. To this end, the researchers employed a quantitative approach. Data were obtained through questionnaires, with a sample of 15 parents having been purposively selected. The findings revealed that parents are unaware that they have to monitor the learner’s cell phone. Another finding was that parents in the 21-century did not support the use of mobile phones in education. The researchers recommend that parent’s discussion forums be created to educate parents on how a cell phone can be used in education. Cellphone companies need to be encouraged to educate parents on how they monitor cell phones used by learners. Another recommendation was that network providers need to restrict access to searching on the internet according to age.Keywords: parental monitoring, app blocking services, learner’s cell phone use, cell phone
Procedia PDF Downloads 1516881 Unexplored Anti-HCV Potential of Lichen rangiferinus: An in Vitro Study over Virus Cultures
Authors: Ila Shukla, Lubna Azmi, Shyam Sunder Gupta, C. V. Rao
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Treatments against Hepatitis-C virus (HCV) are already available, but the current high cost of such treatments limit them to wealthy patients only. Hence our current study is aimed at the rectification of HCV infection by using Lichen rangiferinus (LRE) extract in in vitro cultures. Anti-HCV activity of the given extract was evaluated using the virus grown in cell culture (HCVcc). Two control inhibitors, erlotinib and telaprevir, were systematically included in each experiment. At the end of the incubation period, we evaluated cell viability and viral replication. The LRE inhibited the growth of HCV in a dose dependent manner.Keywords: Erlotinib, Hepatitis C, Lichen rangiferinus, Telaprevir
Procedia PDF Downloads 3296880 Effectuating Theology of Culture: The Only Weapon to Confront 21st Century Global Godless Culture
Authors: Hram Bik
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This is an analytical paper on how to apply theology to the global godless culture. The paper will analyze and materialize theology of culture and come up with theo-cultural principles which will enable Christians to properly engage with today godless culture. If theology and daily life are in any way split apart, Christians will lose the authenticity essential to their calling. Living out godliness in the ungodly culture requires materializing theology into daily life. To do that has become an unbeatable challenge for Christians in 21st century with the overtaking in of global godless culture enforced by Information Technology resulting in rapid and chaotic change of global lifestyles wherein Christianity stands in danger of being swallowed up. Staying away from the culture will rob Christianity of its mission to witness and staying with and like it will rob Christianity of its effectiveness. Thus the question is how should today Christians apply theology to the culture wherein what are said to be sins in the Bible no longer look like sins? Should we forge an all-out war against it or should distance ourselves away from it? The extreme response to it could fruit Christian Jihadism on the right and the apathetic response would let it booming with no one attempting to stop it on the left. This paper calls for global Christians to essentially make theology a part of their daily lives to form a united global force to influence the godless global culture by influencing our own family and community.Keywords: Christians, global culture, godliness, theology
Procedia PDF Downloads 2586879 Contribution of Hydrogen Peroxide in the Selective Aspect of Prostate Cancer Treatment by Cold Atmospheric Plasma
Authors: Maxime Moreau, Silvère Baron, Jean-Marc Lobaccaro, Karine Charlet, Sébastien Menecier, Frédéric Perisse
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Cold Atmospheric Plasma (CAP) is an ionized gas generated at atmospheric pressure with the temperature of heavy particles (molecules, ions, atoms) close to the room temperature. Recent studies have shown that both in-vitro and in-vivo plasma exposition to many cancer cell lines are efficient to induce the apoptotic way of cell death. In some other works, normal cell lines seem to be less impacted by plasma than cancer cell lines. This is called selectivity of plasma. It is highly likely that the generated RNOS (Reactive Nitrogen Oxygen Species) in the plasma jet, but also in the medium, play a key-role in this selectivity. In this study, two CAP devices will be compared to electrical power, chemical species composition and their efficiency to kill cancer cells. A particular focus on the action of hydrogen peroxide will be made. The experiments will take place as described next for both devices: electrical and spectroscopic characterization for different voltages, plasma treatment of normal and cancer cells to compare the CAP efficiency between cell lines and to show that death is induced by an oxidative stress. To enlighten the importance of hydrogen peroxide, an inhibitor of H2O2 will be added in cell culture medium before treatment and a comparison will be made between the results of cell viability in this case and those from a simple plasma exposition. Besides, H2O2 production will be measured by only treating medium with plasma. Cell lines will also be exposed to different concentrations of hydrogen peroxide in order to characterize the cytotoxic threshold for cells and to make a comparison with the quantity of H2O2 produced by CAP devices. Finally, the activity of catalase for different cell lines will be quantified. This enzyme is an important antioxidant agent against hydrogen peroxide. A correlation between cells response to plasma exposition and this activity could be a strong argument in favor of the predominant role of H2O2 to explain the selectivity of plasma cancer treatment by cold atmospheric plasma.Keywords: cold atmospheric plasma, hydrogen peroxide, prostate cancer, selectivity
Procedia PDF Downloads 1416878 Mathematical Modeling of Cell Volume Alterations under Different Osmotic Conditions
Authors: Juliana A. Knocikova, Yann Bouret, Médéric Argentina, Laurent Counillon
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Cell volume, together with membrane potential and intracellular hydrogen ion concentration, is an essential biophysical parameter for normal cellular activity. Cell volumes can be altered by osmotically active compounds and extracellular tonicity. In this study, a simple mathematical model of osmotically induced cell swelling and shrinking is presented. Emphasis is given to water diffusion across the membrane. The mathematical description of the cellular behavior consists in a system of coupled ordinary differential equations. We compare experimental data of cell volume alterations driven by differences in osmotic pressure with mathematical simulations under hypotonic and hypertonic conditions. Implications for a future model are also discussed.Keywords: eukaryotic cell, mathematical modeling, osmosis, volume alterations
Procedia PDF Downloads 4556877 Reproduction of New Media Art Village around NTUT: Heterotopia of Visual Culture Art Education
Authors: Yu Cheng-Yu
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‘Heterotopia’, ‘Visual Cultural Art Education’ and ‘New Media’ of these three subjects seemingly are irrelevant. In fact, there are synchronicity and intertextuality inside. In addition to visual culture, art education inspires students the ability to reflect on popular culture image through visual culture teaching strategies in school. We should get involved in the community to construct the learning environment that conveys visual culture art. This thesis attempts to probe the heterogeneity of space and value from Michel Foucault and to research sustainable development strategy in ‘New Media Art Village’ heterogeneity from Jean Baudrillard, Marshall McLuhan's media culture theory and social construction ideology. It is possible to find a new media group that can convey ‘Visual Culture Art Education’ around the National Taipei University of Technology in this commercial district that combines intelligent technology, fashion, media, entertainment, art education, and marketing network. Let the imagination and innovation of ‘New Media Art Village’ become ‘implementable’ and new media Heterotopia of inter-subjectivity with the engagement of big data and digital media. Visual culture art education will also bring aesthetics into the community by New Media Art Village.Keywords: social construction, heterogeneity, new media, big data, visual culture art education
Procedia PDF Downloads 2376876 Dynamic Thermal Modelling of a PEMFC-Type Fuel Cell
Authors: Marco Avila Lopez, Hasnae Ait-Douchi, Silvia De Los Santos, Badr Eddine Lebrouhi, Pamela Ramírez Vidal
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In the context of the energy transition, fuel cell technology has emerged as a solution for harnessing hydrogen energy and mitigating greenhouse gas emissions. An in-depth study was conducted on a PEMFC-type fuel cell, with an initiation of an analysis of its operational principles and constituent components. Subsequently, the modelling of the fuel cell was undertaken using the Python programming language, encompassing both steady-state and transient regimes. In the case of the steady-state regime, the physical and electrochemical phenomena occurring within the fuel cell were modelled, with the assumption of uniform temperature throughout all cell compartments. Parametric identification was carried out, resulting in a remarkable mean error of only 1.62% when the model results were compared to experimental data documented in the literature. The dynamic model that was developed enabled the scrutiny of the fuel cell's response in terms of temperature and voltage under varying current conditions.Keywords: fuel cell, modelling, dynamic, thermal model, PEMFC
Procedia PDF Downloads 766875 Co-Culture with Murine Stromal Cells Enhances the In-vitro Expansion of Hematopoietic Stem Cells in Response to Low Concentrations of Trans-Resveratrol
Authors: Mariyah Poonawala, Selvan Ravindran, Anuradha Vaidya
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Despite much progress in understanding the regulatory factors and cytokines that support the maturation of the various cell lineages of the hematopoietic system, factors that govern the self-renewal and proliferation of hematopoietic stem cells (HSCs) is still a grey area of research. Hematopoietic stem cell transplantation (HSCT) has evolved over the years and gained tremendous importance in the treatment of both malignant and non-malignant diseases. However, factors such as graft rejection and multiple organ failure have challenged HSCT from time to time, underscoring the urgent need for development of milder processes for successful hematopoietic transplantation. An emerging concept in the field of stem cell biology states that the interactions between the bone-marrow micro-environment and the hematopoietic stem and progenitor cells is essential for regulation, maintenance, commitment and proliferation of stem cells. Understanding the role of mesenchymal stromal cells in modulating the functionality of HSCs is, therefore, an important area of research. Trans-resveratrol has been extensively studied for its various properties to combat and prevent cancer, diabetes and cardiovascular diseases etc. The aim of the present study was to understand the effect of trans-resveratrol on HSCs using single and co-culture systems. We have used KG1a cells since it is a well accepted hematopoietic stem cell model system. Our preliminary experiments showed that low concentrations of trans-resveratrol stimulated the HSCs to undergo proliferation whereas high concentrations of trans-resveratrol did not stimulate the cells to proliferate. We used a murine fibroblast cell line, M210B4, as a stromal feeder layer. On culturing the KG1a cells with M210B4 cells, we observed that the stimulatory as well as inhibitory effects of trans-resveratrol at low and high concentrations respectively, were enhanced. Our further experiments showed that low concentration of trans-resveratrol reduced the generation of reactive oxygen species (ROS) and nitric oxide (NO) whereas high concentrations increased the oxidative stress in KG1a cells. We speculated that perhaps the oxidative stress was imposing inhibitory effects at high concentration and the same was confirmed by performing an apoptotic assay. Furthermore, cell cycle analysis and growth kinetic experiments provided evidence that low concentration of trans-resveratrol reduced the doubling time of the cells. Our hypothesis is that perhaps at low concentration of trans-resveratrol the cells get pushed into the G0/G1 phase and re-enter the cell cycle resulting in their proliferation, whereas at high concentration the cells are perhaps arrested at G2/M phase or at cytokinesis and therefore undergo apoptosis. Liquid Chromatography-Quantitative-Time of Flight–Mass Spectroscopy (LC-Q-TOF MS) analyses indicated the presence of trans-resveratrol and its metabolite(s) in the supernatant of the co-cultured cells incubated with high concentration of trans-resveratrol. We conjecture that perhaps the metabolites of trans-resveratrol are responsible for the apoptosis observed at the high concentration. Our findings may shed light on the unsolved problems in the in vitro expansion of stem cells and may have implications in the ex vivo manipulation of HSCs for therapeutic purposes.Keywords: co-culture system, hematopoietic micro-environment, KG1a cell line, M210B4 cell line, trans-resveratrol
Procedia PDF Downloads 2516874 Safety of Mesenchymal Stem Cells Therapy: Potential Risk of Spontaneous Transformations
Authors: Katarzyna Drela, Miroslaw Wielgos, Mikolaj Wrobel, Barbara Lukomska
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Mesenchymal stem cells (MSCs) have a great potential in regenerative medicine. Since the initial number of isolated MSCs is limited, in vitro propagation is often required to reach sufficient numbers of cells for therapeutic applications. During long-term culture MSCs may undergo genetic or epigenetic alterations that subsequently increase the probability of spontaneous malignant transformation. Thus, factors that influence genomic stability of MSCs following long-term expansions need to be clarified before cultured MSCs are employed for clinical application. The aim of our study was to investigate the potential for spontaneous transformation of human neonatal cord blood (HUCB-MSCs) and adult bone marrow (BM-MSCs) derived MSCs. Materials and Methods: HUCB-MSCs and BM-MSCs were isolated by standard Ficoll gradient centrifugations method. Isolated cells were initially plated in high density 106 cells per cm2. After 48 h medium were changed and non-adherent cells were removed. The malignant transformation of MSCs in vitro was evaluated by morphological changes, proliferation rate, ability to enter cell senescence, the telomerase expression and chromosomal abnormality. Proliferation of MSCs was analyzed with WST-1 reduction method and population doubling time (PDT) was calculated at different culture stages. Then the expression pattern of genes characteristic for mesenchymal or epithelial cells, as well as transcriptions factors were examined by RT-PCR. Concomitantly, immunocytochemical analysis of gene-related proteins was employed. Results: Our studies showed that MSCs from all bone marrow isolations ultimately entered senescence and did not undergo spontaneous malignant transformation. However, HUCB-MSCs from one of the 15 donors displayed an increased proliferation rate, failed to enter senescence, and exhibited an altered cell morphology. In this sample we observed two different cell phenotypes: one mesenchymal-like exhibited spindle shaped morphology and express specific mesenchymal surface markers (CD73, CD90, CD105, CD166) with low proliferation rate, and the second one with round, densely package epithelial-like cells with significantly increased proliferation rate. The PDT of epithelial-like populations was around 1day and 100% of cells were positive for proliferation marker Ki-67. Moreover, HUCB-MSCs showed a positive expression of human telomerase reverse transcriptase (hTERT), cMYC and exhibit increased number of CFU during the long-term culture in vitro. Furthermore, karyotype analysis revealed chromosomal abnormalities including duplications. Conclusions: Our studies demonstrate that HUCB-MSCs are susceptible to spontaneous malignant transformation during long-term culture. Spontaneous malignant transformation process following in vitro culture has enormous effect on the biosafety issues of future cell-based therapies and regenerative medicine regimens.Keywords: mesenchymal stem cells, spontaneous, transformation, long-term culture
Procedia PDF Downloads 2556873 Direct Current Electric Field Stimulation against PC12 Cells in 3D Bio-Reactor to Enhance Axonal Extension
Authors: E. Nakamachi, S. Tanaka, K. Yamamoto, Y. Morita
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In this study, we developed a three-dimensional (3D) direct current electric field (DCEF) stimulation bio-reactor for axonal outgrowth enhancement to generate the neural network of the central nervous system (CNS). By using our newly developed 3D DCEF stimulation bio-reactor, we cultured the rat pheochromocytoma cells (PC12) and investigated the effects on the axonal extension enhancement and network generation. Firstly, we designed and fabricated a 3D bio-reactor, which can load DCEF stimulation on PC12 cells embedded in the collagen gel as extracellular environment. The connection between the electrolyte and the medium using salt bridges for DCEF stimulation was introduced to avoid the cell death by the toxicity of metal ion. The distance between the salt bridges was adopted as the design variable to optimize a structure for uniform DCEF stimulation, where the finite element (FE) analyses results were used. Uniform DCEF strength and electric flux vector direction in the PC12 cells embedded in collagen gel were examined through measurements of the fabricated 3D bio-reactor chamber. Measurement results of DCEF strength in the bio-reactor showed a good agreement with FE results. In addition, the perfusion system was attached to maintain pH 7.2 ~ 7.6 of the medium because pH change was caused by DCEF stimulation loading. Secondly, we disseminated PC12 cells in collagen gel and carried out 3D culture. Finally, we measured the morphology of PC12 cell bodies and neurites by the multiphoton excitation fluorescence microscope (MPM). The effectiveness of DCEF stimulation to enhance the axonal outgrowth and the neural network generation was investigated. We confirmed that both an increase of mean axonal length and axogenesis rate of PC12, which have been exposed 5 mV/mm for 6 hours a day for 4 days in the bioreactor. We found following conclusions in our study. 1) Design and fabrication of DCEF stimulation bio-reactor capable of 3D culture nerve cell were completed. A uniform electric field strength of average value of 17 mV/mm within the 1.2% error range was confirmed by using FE analyses, after the structure determination through the optimization process. In addition, we attached a perfusion system capable of suppressing the pH change of the culture solution due to DCEF stimulation loading. 2) Evaluation of DCEF stimulation effects on PC12 cell activity was executed. The 3D culture of PC 12 was carried out adopting the embedding culture method using collagen gel as a scaffold for four days under the condition of 5.0 mV/mm and 10mV/mm. There was a significant effect on the enhancement of axonal extension, as 11.3% increase in an average length, and the increase of axogenesis rate. On the other hand, no effects on the orientation of axon against the DCEF flux direction was observed. Further, the network generation was enhanced to connect longer distance between the target neighbor cells by DCEF stimulation.Keywords: PC12, DCEF stimulation, 3D bio-reactor, axonal extension, neural network generation
Procedia PDF Downloads 1826872 Comparison of Physical and Chemical Effects on Senescent Cells
Authors: Svetlana Guryeva, Inna Kornienko, Andrey Usanov, Dmitry Usanov, Elena Petersen
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Every day cells in our organism are exposed to various factors: chemical agents, reactive oxygen species, ionizing radiation, and others. These factors can cause damage to DNA, cellular membrane, intracellular compartments, and proteins. The fate of cells depends on the exposure intensity and duration. The prolonged and intense exposure causes the irreversible damage accumulation, which triggers the permanent cell cycle arrest (cellular senescence) or cell death programs. In the case of low dose of impacts, it can lead to cell renovation and to cell functional state improvement. Therefore, it is a pivotal question to investigate the factors and doses that result in described positive effects. In order to estimate the influence of different agents, the proliferation index and levels of cell death markers (annexin V/propidium iodide), senescence-associated β-galactosidase, and lipofuscin were measured. The experiments were conducted on primary human fibroblasts of the 8th passage. According to the levels of mentioned markers, these cells were defined as senescent cells. The effect of low-frequency magnetic field was investigated. Different modes of magnetic field exposure were tested. The physical agents were compared with chemical agents: metformin (10 mM) and taurine (0.8 mM and 1.6 mM). Cells were incubating with chemicals for 5 days. The highest decrease in the level of senescence-associated β-galactosidase (21%) and lipofuscin (17%) was observed in the primary senescent fibroblasts after 5 days after double treatments with 48 h intervals with low-frequency magnetic field. There were no significant changes in the proliferation index after magnetic field application. The cytotoxic effect of magnetic field was not observed. The chemical agent taurine (1.6 mM) decreased the level of senescence-associated β-galactosidase (23%) and lipofuscin (22%). Metformin improved the activity of senescence-associated β-galactosidase on 15% and the level of lipofuscin on 19% in this experiment. According to these results, the effect of double treatment with 48 h interval with low-frequency magnetic field and the effect of taurine (1.6 mM) were comparable to the effect of metformin, for which anti-aging properties are proved. In conclusion, this study can become the first step towards creation of the standardized system for the investigation of different effects on senescent cells.Keywords: biomarkers, magnetic field, metformin, primary fibroblasts, senescence, taurine
Procedia PDF Downloads 2696871 An Innovative Poly System Theory for the Go-Out of Chinese Culture
Authors: Jianhua Wang, Ying Zhou, Han Guo
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Translation underwent culture turn for more than half a century, which brought translation and its studies beyond intra-texts. Different cultures in recent years have developed towards a translation turn, which made a great contribution to relocate national or local cultures being localized to become regional or global cultures. As China grows quickly economically integrating into the world, it becomes urgent to relate China’s story and disseminate the Chinese culture. Due to the weaknesses and drawbacks of different existing cultural translation theories for Chinese culture to go out, a new perspective on translation turn for the go-out of Chinese culture should be drawn to spread better and disseminate Chinese culture to other countries. Based on the existing cultural translation theories, the equivalence of ideology, style of the translator and agency of the support are proposed to draw a new perspective: an innovative poly-system theory for Chinese culture translation.Keywords: cultural translation theory, Chinese culture, innovative poly system, global cultures
Procedia PDF Downloads 4456870 Microfluidic Fluid Shear Mechanotransduction Device Using Linear Optimization of Hydraulic Channels
Authors: Sanat K. Dash, Rama S. Verma, Sarit K. Das
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A logarithmic microfluidic shear device was designed and fabricated for cellular mechanotransduction studies. The device contains four cell culture chambers in which flow was modulated to achieve a logarithmic increment. Resistance values were optimized to make the device compact. The network of resistances was developed according to a unique combination of series and parallel resistances as found via optimization. Simulation results done in Ansys 16.1 matched the analytical calculations and showed the shear stress distribution at different inlet flow rates. Fabrication of the device was carried out using conventional photolithography and PDMS soft lithography. Flow profile was validated taking DI water as working fluid and measuring the volume collected at all four outlets. Volumes collected at the outlets were in accordance with the simulation results at inlet flow rates ranging from 1 ml/min to 0.1 ml/min. The device can exert fluid shear stresses ranging four orders of magnitude on the culture chamber walls which will cover shear stress values from interstitial flow to blood flow. This will allow studying cell behavior in the long physiological range of shear stress in a single run reducing number of experiments.Keywords: microfluidics, mechanotransduction, fluid shear stress, physiological shear
Procedia PDF Downloads 1266869 Nationalist Approach to the Music Culture in Early Republic Period in Turkey
Authors: Hilmi Yazici
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Just after Ottoman period, new more homogenic republic was struggling to form a national identity and dealing with the cultural and historical background of the nation. This new republic had an aim of modernization and westernization which started in the late ottoman period. In this process, the culture was an important basis to form a new nation and it clearly put forward that the new citizens of the new national republic are to have a modern and national culture. The result of this aimed change was to find the Turkish culture suppressed among the common people of the Anatolia and to take the western modernization and breed this with national culture. So in this context, we can say that this approach separated the people from ottoman culture and its roots to empower the national identity. Repeatedly, it may be said that Turkish folkloric music was an important basis for the new revolution, on the other hand classical Turkish music was alienated with the idea that it didn’t belong to Turkish culture. So the aim of this study is to determine how these efforts to nationalize a new identity and culture was successful and conscious intervention to folkloric Turkish music became efficient.Keywords: opera, nationalism in music, Turkish music
Procedia PDF Downloads 2856868 Experimental Investigation of the Impact of Biosurfactants on Residual-Oil Recovery
Authors: S. V. Ukwungwu, A. J. Abbas, G. G. Nasr
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The increasing high price of natural gas and oil with attendant increase in energy demand on world markets in recent years has stimulated interest in recovering residual oil saturation across the globe. In order to meet the energy security, efforts have been made in developing new technologies of enhancing the recovery of oil and gas, utilizing techniques like CO2 flooding, water injection, hydraulic fracturing, surfactant flooding etc. Surfactant flooding however optimizes production but poses risk to the environment due to their toxic nature. Amongst proven records that have utilized other type of bacterial in producing biosurfactants for enhancing oil recovery, this research uses a technique to combine biosurfactants that will achieve a scale of EOR through lowering interfacial tension/contact angle. In this study, three biosurfactants were produced from three Bacillus species from freeze dried cultures using sucrose 3 % (w/v) as their carbon source. Two of these produced biosurfactants were screened with the TEMCO Pendant Drop Image Analysis for reduction in IFT and contact angle. Interfacial tension was greatly reduced from 56.95 mN.m-1 to 1.41 mN.m-1 when biosurfactants in cell-free culture (Bacillus licheniformis) were used compared to 4. 83mN.m-1 cell-free culture of Bacillus subtilis. As a result, cell-free culture of (Bacillus licheniformis) changes the wettability of the biosurfactant treatment for contact angle measurement to more water-wet as the angle decreased from 130.75o to 65.17o. The influence of microbial treatment on crushed rock samples was also observed by qualitative wettability experiments. Treated samples with biosurfactants remained in the aqueous phase, indicating a water-wet system. These results could prove that biosurfactants can effectively change the chemistry of the wetting conditions against diverse surfaces, providing a desirable condition for efficient oil transport in this way serving as a mechanism for EOR. The environmental friendly effect of biosurfactants applications for industrial purposes play important advantages over chemically synthesized surfactants, with various possible structures, low toxicity, eco-friendly and biodegradability.Keywords: bacillus, biosurfactant, enhanced oil recovery, residual oil, wettability
Procedia PDF Downloads 2776867 Creative Culture to Innovative Culture: Transformal Operation
Authors: Peer M. Sathikh
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Creativity and innovation have become an important phenomenon today, whose potential is being realized through the success of Apple, Google/Android, Nike, Virgin, Dyson and other multinationals that are a household name today. Creativity and Innovation are, many times, used interchangeably, causing confusion as to what each represents and are capable of. Attempts to understand creativity and innovation clearly point to the difference, and at the same time, inter-dependency of one on the other. The assumption that having more creative personnel in a team will translate into innovation sooner or later seems generally counterproductive. What helps define the role of creativity and innovation in an organization and how can one build an innovative team? This paper points to the importance of understanding creative culture and innovation culture in order to bring about the desired innovation outcome and proposes a means to transform one to another as ideas move from mere ideas to useful innovation.Keywords: creativity, innovation, creative culture, innovation culture, transformal operators
Procedia PDF Downloads 3966866 Aligning Organizational Culture and Compensation Strategies
Authors: Giuseppe Maria Russo, Patrícia Amélia Tomei, Antônio Linhares, André Moreira Santos
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Alignment between management strategies, policies and practices with organizational cultures holds great potential to meet the challenges of retaining professionals and maintaining their commitment. In this article, authors consider that when it is aligned with company strategy, compensation acts as an incentive for developing common visions within the organizational culture. This article verified the correlation between types of culture and compensation’s strategic components and provided inputs for the definition of strategies aligned with cultural typologies. We conclude that the impact of compensation variables varies according to the type of organizational culture. This result reinforces the theory that different cultures define different organizational strategies. Thus, compensation strategies may explain types of organizational culture.Keywords: compensation, Handy’s cultural typology, organizational culture, rewards
Procedia PDF Downloads 6566865 A Guidance to Enhance the Risk Culture among the Organizations
Authors: Najeebah Almahmeed
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Risk Management is an evolving subject among organizations that include corporations, governments, non-governmental organizations, and not-for-profit corporations. In order to enhance awareness around the importance of Risk Management and make sure everyone is using it in their day-to-day job, the Risk Culture topic has emerged and gained importance not only in the Finance Sector but also in the National Oil Companies in Kuwait. Risk Culture can be defined as the shared beliefs, attitudes, and behaviors within a company that guide its approach to managing risks. It acts as a connecting force that links policies, procedures, and individuals, influencing how risks are understood and tackled through activities. In this research, benefits of Risk Culture are shared, guidelines are presented to promote a risk aware culture, and fully embed and enforce Risk-based processes and procedures. Moreover, this research demonstrates methodologies of measuring the Risk Culture using specific dimensions and clusters.Keywords: clusters, dimensions, national oil companies, risk culture, risk management
Procedia PDF Downloads 716864 Influence of Preheating Self-Adhesive Cements on the Degree of Conversion, Cell Migration and Cell Viability in NIH/3T3
Authors: Celso Afonso Klein Jr., Henrique Cantarelli, Fernando Portella, Keiichi Hosaka, Eduardo Reston, Fabricio Collares, Roberto Zimmer
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TTo evaluate the influence of preheating self-adhesive cement at 39ºC on cell migration, cytotoxicity and degree of conversion. RelyX U200, Set PP and MaxCem Elite were subjected to a degree of conversion analysis (FTIR-ATR). For the cytotoxicity analysis, extracts (24 h and 7 days) were placed in contact with NIH/3T3 cells. For cell migration, images were captured of each sample until the possible closure of the cleft occurred. In the results of the degree of conversion, preheating did not improve the conversion of cement. For the MTT, preheating did not improve the results within 24 hours. However, it generated positive results within 7 days for the Set PP resin cement. For cell migration, high rates of cell death were found in all groups. It is concluded that preheating at 39ºC caused a positive effect only in increasing the cell viability of the Set PP resin cement and that both materials analyzed are highly cytotoxic.Keywords: dental cements, resin cements, degree of conversion, cytotoxicity, cell migration assays
Procedia PDF Downloads 616863 Rauvolfine B Isolated from the Bark of Rauvolfia reflexa (Apocynaceae) Induces Apoptosis through Activation of Caspase-9 Coupled with S Phase Cell Cycle Arrest
Authors: Mehran Fadaeinasab, Hamed Karimian, Najihah Mohd Hashim, Hapipah Mohd Ali
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In this study, three indole alkaloids namely; rauvolfine B, macusine B, and isoreserpiline have been isolated from the dichloromethane crude extract of Rauvolfia reflexa bark (Apocynaceae). The structural elucidation of the isolated compounds has been performed using spectral methods such as UV, IR, MS, 1D, and 2D NMR. Rauvolfine B showed anti proliferation activity on HCT-116 cancer cell line, its cytotoxicity induction was observed using MTT assay in eight different cell lines. Annexin-V is serving as a marker for apoptotic cells and the Annexin-V-FITC assay was carried out to observe the detection of cell-surface Phosphatidylserine (PS). Apoptosis was confirmed by using caspase-8 and -9 assays. Cell cycle arrest was also investigated using flowcytometric analysis. rauvolfine B had exhibited significantly higher cytotoxicity against HCT-116 cell line. The treatment significantly arrested HCT-116 cells in the S phase. Together, the results presented in this study demonstrated that rauvolfine B inhibited the proliferation of HCT-116 cells and programmed cell death followed by cell cycle arrest.Keywords: apocynacea, indole alkaloid, apoptosis, cell cycle arrest
Procedia PDF Downloads 3266862 Cultural Traditions Petik Laut and Onjem in Gili Island, Indonesia That Potential as Ecotourism to Bring Indonesia's Culture to the World
Authors: Dwi Yulian Fahruddin Shah, Mochammad Luthfy Rizaldy Dwi Putra, Tommy Adi Rachmawan, Mona Annisa Matondang, Nadya Sylvia, Hilmy Ramzy Rinaldy
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Gili island is one of the island in Indonesia which is located in Probolinggo city, East Java. Gili Island has some potential culture as local wisdom that can be used as tourism commodity because it can be used as attractive ecotourism. With the ecotourism that utilize local wisdom of Indonesian’s culture that located in Gili can introduce the richness of Indonesian culture in the world that will increase foreign exchange. One of the cultural potential as local wisdom in Gili island are Petik Laut and Onjem. It are a culture in Gili island that can’t be found in other island in Indonesia. Not just that but also it are a cultural identity that is owned by Gili island which has fill the criteria to be used as local wisdom that can be used as ecotourism that can bring Indonesian culture to the world so that the tourists of the world will visit to Indonesia, especially to Gili island to see Petik Laut and Onjem culture directly.Keywords: Gili island, petik laut and onjem culture, ecotourism, indonesia’s culture
Procedia PDF Downloads 5496861 Digital Storytelling for Community Culture
Authors: Sariyapa Kantawan, Muanfun Kongsomsawaeng
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Chanthaburi River community is an old mixed-culture village established in the 16th century. The town advanced more rapidly than others due to the ease of transportation at the time, which used the river as a road. Therefore, the province's first road begins here, propelling it to become an important commercial and trading center for almost a century. As a result of diverse culture, the architecture has been affected by Western, Thai, Chinese, and Vietnamese, resulting in a new and distinctive style. To share the realm of memory, digital media enable the city to communicate its history and culture. This article describes a project that combines the concepts of digital storytelling and augmented reality and connects them to Chanthaburi River Community Culture by using QR codes as makers to display 3D models on mobile screens.Keywords: digital storytelling, community culture, river community, cultural heritage, augmented reality
Procedia PDF Downloads 496860 Building Organisational Culture That Stimulates Creativity and Innovation
Authors: Ala Hanetite
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The purpose of this article is to present, by means of a model, the determinants of organisational culture which influence creativity and innovation. A literature study showed that a model, based on the open systems theory and the work of Schein, can offer a holistic approach in describing organisational culture. The relationship between creativity, innovation and culture is discussed in this context. Against the background of this model, the determinants of organisational culture were identified. The determinants are strategy, structure, support mechanisms, behaviour that encourages innovation, and open communication. The influence of each determinant on creativity and innovation is discussed. Values, norms and beliefs that play a role in creativity and innovation can either support or inhibit creativity and innovation depending on how they influence individual and group behaviour. This is also explained in the article.Keywords: attitudes, creativity, innovation, organisational culture
Procedia PDF Downloads 5836859 Role of Micro-Patterning on Stem Cell-Material Interaction Modulation and Cell Fate
Authors: Lay Poh Tan, Chor Yong Tay, Haiyang Yu
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Micro-contact printing is a form of soft lithography that uses the relief patterns on a master polydimethylsiloxane (PDMS) stamp to form patterns of self-assembled monolayers (SAMs) of ink on the surface of a substrate through conformal contact technique. Here, we adopt this method to print proteins of different dimensions on our biodegradable polymer substrates. We started off with printing 20-500 μm scale lanes of fibronectin to engineer the shape of bone marrow derived human mesenchymal stem cell (hMSCs). After 8 hours of culture, the hMSCs adopted elongated shapes, and upon analysis of the gene expressions, genes commonly associated with myogenesis (GATA-4, MyoD1, cTnT and β-MHC) and neurogenesis (NeuroD, Nestin, GFAP, and MAP2) were up-regulated but gene expression associated to osteogenesis (ALPL, RUNX2, and SPARC) were either down modulated or remained at the nominal level. This is the first evidence that cellular morphology control via micropatterning could be used to modulate stem cell fate without external biochemical stimuli. We further our studies to modulate the focal adhesion (FA) instead of the macro shape of cells. Micro-contact printed islands of different smaller dimensions were investigated. We successfully regulated the FAs into dense FAs and elongated FAs by micropatterning. Additionally, the combined effects of hard (40.4 kPa), and intermediate (10.6 kPa) PA gel and FAs patterning on hMSCs differentiation were studied. Results showed that FA and matrix compliance plays an important role in hMSCs differentiation, and there is a cross-talk between different physical stimulants and the significance of these stimuli can only be realized if they are combined at the optimum level.Keywords: micro-contact printing, polymer substrate, cell-material interaction, stem cell differentiation
Procedia PDF Downloads 168