Search results for: cell free supernatant

Authors: Venkat Garigapati

Abstract:

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 72

Authors: Ipsita Biswas, Arnab Sarkar, Ashikur Rahaman, Gopeswar Mukherjee, Subhrangsu Chatterjee, Shamee Bhattacharjee, Deba Prasad Mandal

Abstract:

One of the major reasons for the high mortality rate of lung cancer is the substantial delays in disease detection at late metastatic stages. It is of utmost importance to understand the detailed molecular signaling and detect the molecular markers that can be used for the early diagnosis of cancer. Several studies explored the emerging roles of long noncoding RNAs (lncRNAs) in various cancers as well as lung cancer. A long non-coding RNA LINC00273 was recently discovered to promote cancer cell migration and invasion, and its positive correlation with the pathological stages of metastasis may prove it to be a potential target for inhibiting cancer cell metastasis. Comparing real-time expression of LINC00273 in various human clinical cancer tissue samples with normal tissue samples revealed significantly higher expression in cancer tissues. This long intergenic noncoding RNA was found to be highly expressed in human liver tumor-initiating cells, human gastric adenocarcinoma AGS cell line, as well as human non-small cell lung cancer A549 cell line. SiRNA and shRNA-induced knockdown of LINC00273 in both in vitro and in vivo nude mice significantly subsided AGS and A549 cancer cell migration and invasion. LINC00273 knockdown also reduced TGF-β induced SNAIL, SLUG, VIMENTIN, ZEB1 expression, and metastasis in A549 cells. Plenty of reports have suggested the role of microRNAs of the miR200 family in reversing epithelial to mesenchymal transition (EMT) by inhibiting ZEB transcription factors. In this study, hsa-miR-200a-3p was predicted via IntaRNA-Freiburg RNA tools to be a potential target of LINC00273 with a negative free binding energy of −8.793 kcal/mol, and this interaction was verified as a confirmed target of LINC00273 by RNA pulldown, real-time PCR and luciferase assay. Mechanistically, LINC00273 accelerated TGF-β induced EMT by sponging hsa-miR-200a-3p which in turn liberated ZEB1 and promoted prometastatic functions in A549 cells in vitro as verified by real-time PCR and western blotting. The similar expression patterns of these EMT regulatory pathway molecules, viz. LINC00273, hsa-miR-200a-3p, ZEB1 and TGF-β, were also detected in various clinical samples like breast cancer tissues, oral cancer tissues, lung cancer tissues, etc. Overall, this LINC00273 mediated EMT regulatory signaling can serve as a potential therapeutic target for the prevention of lung cancer metastasis.

Keywords: epithelial to mesenchymal transition, long noncoding RNA, microRNA, non-small-cell lung carcinoma

Procedia PDF Downloads 132

Authors: T. Scattolin, E. Cavarzerani, F. Visentin, F. Rizzolio

Abstract:

Organopalladium compounds have recently attracted attention for their high stability even under physiological conditions and, above all, for their remarkable in vitro cytotoxicity towards cisplatin-resistant cell lines. Among the organopalladium derivatives, those bearing at least one N-heterocyclic carbene ligand (NHC) and the Pd(II)-η³-allyl fragment have exhibited IC₅₀ values in the micro and sub-micromolar range towards several cancer cell lines in vitro and in some cases selectivity towards cancerous vs. non-tumorigenic cells. Herein, a selection of allyl and indenyl palladates were synthesized using a solvent-free method consisting of grinding the corresponding palladium precursors with different saturated and unsaturated azolium salts. All compounds have been fully characterized by NMR, XRD and elemental analyses. The intramolecular H, Cl interaction has been elucidated and quantified using the Voronoi Deformation Density scheme. Most of the complexes showed excellent cytotoxicity towards ovarian cancer cell lines, with I₅₀ values comparable to or even lower than cisplatin. Interestingly, the potent anticancer activity was also confirmed in a high-serous ovarian cancer (HGSOC) patient-derived tumoroid, with a clear superiority of this class of compounds over classical platinum-based agents. Finally, preliminary enzyme inhibition studies of the synthesized palladate complexes against the model TrxR show that the compounds have high activity comparable to or even higher than auranofin and classical Au(I) NHC complexes. Based on such promising data, further in vitro and in vivo experiments and in-depth mechanistic studies are ongoing in our laboratories.

Keywords: anticancer activity, palladium complexes, organoids, indenyl and allyl ligands

Procedia PDF Downloads 70

Authors: Leila Ayat, Afak Meftah

Abstract:

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 486

Authors: Rajapaksha Gedara Prasad Tharanga Jayasooriya, Matharage Gayani Dilshara, Yung Hyun Choi, Gi-Young Kim

Abstract:

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 412

Authors: Aleksandrs Kovalcuks

Abstract:

Egg yolk oil is a natural source of bioactive compounds such as unsaturated fatty acids, oil soluble vitamins, pigments and others. Bioactive compound content in egg yolk oil depends from its content in eggs, from which oil was extracted. Many studies show that bioactive compound content in egg is correlated to the content of these compounds in hen feed, but there is also an opinion that hen housing systems also have influence on egg chemical content. The aim of this study was to determine which factor, laying hen housing system or hen diet, has a primary influence on bioactive compound content in egg yolk oil. The egg yolk oil was extracted from eggs obtained from 4 different hen housing systems: cage, barn and two groups of free range. All hens were fed with commercially produced compound feed except one group of free range hens which get free diet – pastured hens. Extracted egg yolk oils were analyzed for fatty acids, oil soluble vitamins and β-carotene content. α-tocopherol, ergocalcipherol and polyunsaturated fatty acid content in egg yolk oil was higher from eggs obtained from all housing systems where hens were fed with commercial compound feed. β-carotene and retinol content in egg yolk oils from free range free diet eggs was significantly (p>0.05) higher that from other eggs because hens have access to green forage. Hen physical activity in free range housing systems decreases content of some bioactive compound in egg yolk oil.

Keywords: egg yolk oil, vitamins, caged eggs, free range

Procedia PDF Downloads 439

Authors: Sharon Yunger, Liat Altman, Yuval Garini, Yaron Shav-Tal

Abstract:

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 263

Authors: Yessica Nataliani, Miin-Shen Yang

Abstract:

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 375

Authors: Hend Mesbah, Yehia Youssef, Ibrahim Hassan, Shaaban Nosier, Ahmed El-Shazly, Ahmed Helal

Abstract:

The problem of drinking water shortage is becoming more crucial nowadays as a result of the increased demand due to the population growth and the rise in the standard living. In recent years, desalination using electrodialysis powered by solar energy (PV-ED) is being widely used to help provide treated water and reduce the scarcity in water supply. In the present study, a water desalination laboratory scale ED cell with a fixed bed circulation system was designed, developed, and tested. The effect of three parameters (namely, cell voltage , flowrate, and salt concentration) on the removal percentage of salt ions was studied. The cell voltage was adjusted at 3 , 4 and 6 V. A flow rate of 5, 10, and 20 ml/s and an initial salt concentration of 2000, 5000, and 7000 ppm were investigated. The maximum salt percentage removal obtained was 52.5% at the lowest initial concentration (2000 ppm) and at the highest cell voltage (6 V). There was no significant effect of the flow rate on the removal percentage. A model of PV module has also been developed to calculate the dimensions of a solar cell based on the amount of energy consumed and it was calculated from the Overall ED cell voltage.

Keywords: desalination, electrodialysis, solar desalination, photovoltaic electrodialysis

Procedia PDF Downloads 117

Authors: Sana Bembli, Nahla Khraief Haddad, Safya Belghith

Abstract:

This paper deals with a robust model free terminal sliding mode with gravity compensation approach used to control an exoskeleton-upper limb system. The considered system is a 2-DoF robot in interaction with an upper limb used for rehabilitation. The aim of this paper is to control the flexion/extension movement of the shoulder and the elbow joints in presence of matched disturbances. In the first part, we present the exoskeleton-upper limb system modeling. Then, we controlled the considered system by the model free terminal sliding mode with gravity compensation. A stability study is realized. To prove the controller performance, a robustness analysis was needed. Simulation results are provided to confirm the robustness of the gravity compensation combined with to the Model free terminal sliding mode in presence of uncertainties.

Keywords: exoskeleton- upper limb system, model free terminal sliding mode, gravity compensation, robustness analysis

Procedia PDF Downloads 117

Authors: Abhishek Gandhi, Naresh Bhatnagar

Abstract:

In this article, a new technique has been developed to manufacture open cell extruded thermoplastic foamed sheets with the aid of extrudate surface-quenching phenomenon. As the extrudate foam exits the die, its surface is rapidly quenched which results in freezing of cells on the surface, while the cells at the core continue to grow and leads to development of open-cellular microstructure at the core. Influence of chill roll temperature was found to be extremely significant in developing porous morphological attributes. Subsequently, synergistic effect of blowing agent content and chill roll temperature was examined for their expansion ratio and open-cell microstructure. Further, chill roll rotating speed was found extremely significant in obtaining open-cellular foam structures. This study intends to enhance the understanding of researchers working in the area of open-cell foam processing.

Keywords: foams, porous materials, morphology, composite, microscopy, open-cell foams

Procedia PDF Downloads 422

Authors: Win Nee Phong, Tau Chuan Ling, Pau Loke Show

Abstract:

From an industrial perspective, the exploitation of microalgae for protein source is of great economical and commercial interest due to numerous attractive characteristics. Nonetheless, the release of protein from microalgae is limited by the multiple layers of the rigid thick cell wall that generally contain a large proportion of cellulose. Thus an efficient cell disruption process is required to rupture the cell wall. The conventional downstream processing methods which typically involve several unit operational steps such as disruption, isolation, extraction, concentration and purification are energy-intensive and costly. To reduce the overall cost and establish a feasible technology for the success of the large-scale production, microalgal industry today demands a more cost-effective and eco-friendly technique in downstream processing. One of the main challenges to extract the proteins from microalgae is the presence of rigid cell wall. This study aims to provide some guidance on the selection of the efficient solvent to facilitate the proteins released during the cell disruption process. The effects of solvent types such as methanol, ethanol, 1-propanol and water in rupturing the microalgae cell wall were studied. It is interesting to know that water is the most effective solvent to recover proteins from microalgae and the cost is cheapest among all other solvents.

Keywords: green, microalgae, protein, solvents

Procedia PDF Downloads 229

Authors: Chayma Bahar, Chokri Baccouch, Hedi Sakli, Nizar Sakli

Abstract:

Harvesting and transport of optical and radiofrequency signals are a topical subject with multiple challenges. In this paper, we present a Optical RECTENNA system. We propose here a hybrid system solar cell antenna for 5G mobile communications networks. Thus, we propose rectifying circuit. A parametric study is done to follow the influence of load resistance and input power on Optical RECTENNA system performance. Thus, we propose a solar cell antenna structure in the frequency band of future 5G standard in 2.45 GHz bands.

Keywords: antenna, IoT, optical rectenna, solar cell

Procedia PDF Downloads 152

Authors: Mousa Meratizaman, Sina Monadizadeh, Majid Amidpour

Abstract:

One of the most favorable thermal desalination methods used widely today is Multi Effect Desalination. High energy consumption in this method causes coupling it with high temperature power cycle like gas turbine. This combination leads to higher energy efficiency. One of the high temperature power systems which have cogeneration opportunities is Solid Oxide Fuel Cell / Gas Turbine. Integration of Multi Effect Desalination with Solid Oxide Fuel Cell /Gas Turbine power cycle in a range of 300-1000 kW is considered in this article. The exhausted heat of Solid Oxide Fuel Cell /Gas Turbine power cycle is used in Heat Recovery Steam Generator to produce needed motive steam for Desalination unit. Thermodynamic simulation and parametric studies of proposed system are carried out to investigate the system performance.

Keywords: solid oxide fuel cell, thermodynamic simulation, multi effect desalination, gas turbine hybrid cycle

Procedia PDF Downloads 344

Authors: Ratchaneewan Maneemaroj, Paveena Noisuwan, Chonlada Lakhonphon

Abstract:

The smoking is one of the major risk factors in Non-Communicable Disease. Free radicals from cigarette smoke can cause oxidative stress. The oxidative insults can lead to red blood cell (RBC) senescence and are involved in the clearance of red blood cells. The objective of the present study is to assess the association between smoke, oxidative stress evaluated with serum Malondialdehyde (MDA) level and phosphatidylserine (PS) externalization (biomarker of RBC senescence) evaluated with annexin V binding. A total of sixty-four male volunteers aged 25-60 years old were recruited in this study. MDA was measured by colorimetric method. Annexin V binding was detected by flow cytometry. Our results show that there was a significant increase in MDA levels in cigarette smokers as compared to non-smokers (p < 0.001). However, there was no significant different between annexin V binding (% gate) in cigarette smokers and non-smokers (p = 0.978). These results provide evidence of free radical from smoking is associated with oxidative damage to erythrocytes. However, our results suggest that PS externalization is unlikely to have a role in RBC senescence pathway of stressed erythrocytes from cigarette smoke. The other biomarker of RBC senescence should be determined on cigarette smoker erythrocytes.

Keywords: malondialdehyde, phosphatidylserine, RBC senescence, annexin V

Procedia PDF Downloads 408

Authors: Kumari Rekha, Mathur K Dhananjay, Maheshwari Deepanshu, Nautiyal Nidhi, Shubham Smriti, Laal Deepika, Sinha Swati, Kumar Anupam, Biswas Subhrajit, Shiv Kumar Sarin

Abstract:

Background: Mesenchymal stem cells are multipotent stem cells derived from mesoderm and are used for therapeutic purposes because of their self-renewal, homing capacity, Immunomodulatory capability, low immunogenicity and mitochondrial transfer signaling. MSCs have the ability to regulate the mechanism of both innate as well as adaptive immune responses through the modulation of cellular response and the secretion of inflammatory mediators. Different sources of MSC are UC MSC, BM MSC, Dental Pulp, and Adipose MSC. The most frequent source used is umbilical cord tissue due to its being easily available and free of limitations of collection procedures from respective hospitals. The immunosuppressive role of MSCs is particularly interesting for clinical use since it confers resistance to rejection by the host immune response. Methodology: In this study, T helper cells (TH4), Cytotoxic T cells (CD-8), immunoregulatory cells (CD25 +FOXP3+) are compared from isolated MSC from three different methods, UC Dissociation Kit (Miltenyi), Explant Culture and Collagenase Type-IV. To check the immunomodulatory property, these MSCs were seeded with PBMC(Coculture) in CD3 coated 24 well plates. Cd28 antibody was added in coculture for six days. The coculture was analyzed in FACS Verse flow cytometry. Results: From flow cytometry analysis of coculture, it found that All over T helper cells (CD4+) number p<0.0264 increases in (All Enzymes) MSC rather than explant MSC(p>0.0895) as compared to Collagenase(p>0.7889) in a coculture of Activated T cell and Mesenchymal Stem Cell. Similar T reg cells (CD25+, FOXP3+) expression p<0.0234increases in All Enzymes), decreases in Explant and Collagenase. Experiments have shown that MSCs can also directly prevent the cytotoxic activity of CD8 lymphocytes mainly by blocking their proliferation rather than by inhibiting the cytotoxic effect. And promoting the t-reg cells, which helps in the mediation of immune response in various diseases. Conclusion: MSC suppress Cytotoxic CD8 T cell and Enhance immunoregulatory T reg (CD4+, CD25+, FOXP3+) Cell expression. Thus, MSC maintains a proper balance(ratio) between CD4 T cells and Cytotoxic CD8 T cells.

Keywords: MSC, disease, T cell, T regulatory

Procedia PDF Downloads 80

Authors: Jing Zhang, Yvonne Reinwald, Nick Poulson, Alicia El Haj, Chung See, Mike Somekh, Melissa Mather

Abstract:

Biomedical imaging of native tissue using light offers the potential to obtain excellent structural and functional information in a non-invasive manner with good temporal resolution. Image contrast can be derived from intrinsic absorption, fluorescence, or scatter, or through the use of extrinsic contrast. A major challenge in applying optical microscopy to in vivo tissue imaging is the effects of light attenuation which limits light penetration depth and achievable imaging resolution. Recently Selective Plane Illumination Microscopy (SPIM) has been used to map the 3D distribution of fluorophores dispersed in biological structures. In this approach, a focused sheet of light is used to illuminate the sample from the side to excite fluorophores within the sample of interest. Images are formed based on detection of fluorescence emission orthogonal to the illumination axis. By scanning the sample along the detection axis and acquiring a stack of images, 3D volumes can be obtained. The combination of rapid image acquisition speeds with the low photon dose to samples optical sectioning provides SPIM is an attractive approach for imaging biological samples in 3D. To date all implementations of SPIM rely on the use of fluorescence reporters be that endogenous or exogenous. This approach has the disadvantage that in the case of exogenous probes the specimens are altered from their native stage rendering them unsuitable for in vivo studies and in general fluorescence emission is weak and transient. Here we present for the first time to our knowledge a label-free implementation of SPIM that has downstream applications in the clinical setting. The experimental set up used in this work incorporates both label-free and fluorescent illumination arms in addition to a high specification camera that can be partitioned for simultaneous imaging of both fluorescent emission and scattered light from intrinsic sources of optical contrast in the sample being studied. This work first involved calibration of the imaging system and validation of the label-free method with well characterised fluorescent microbeads embedded in agarose gel. 3D constructs of mammalian cells cultured in agarose gel with varying cell concentrations were then imaged. A time course study to track cell proliferation in the 3D construct was also carried out and finally a native tissue sample was imaged. For each sample multiple images were obtained by scanning the sample along the axis of detection and 3D maps reconstructed. The results obtained validated label-free SPIM as a viable approach for imaging cells in a 3D gel construct and native tissue. This technique has the potential use in a near-patient environment that can provide results quickly and be implemented in an easy to use manner to provide more information with improved spatial resolution and depth penetration than current approaches.

Keywords: bioimaging, optics, selective plane illumination microscopy, tissue imaging

Procedia PDF Downloads 223

Authors: Raju Ranjha, Surbhi Aggarwal

Abstract:

Background: Inflammation plays an important role in infectious and non-infectious diseases. MiRNA is also reported to play role in inflammation and associated cancers. Chemokine CXCL12 is also known to play role in inflammation and various cancers. CXCL12/CXCR4 chemokine axis was involved in pathogenesis of IBD specially UC. Supplementation of CXCL12 induces homing of dendritic cells to spleen and enhances control of plasmodium parasite in BALB/c mice. We looked at the regulation of CXCL12β by miRNA in UC colitis. Prolonged inflammation of colon in UC patient increases the risk of developing colorectal cancer. We looked at the expression differences of CXCl12β and its targeting miRNA in cancer susceptible area of colon of UC patients. Aim: Aim of this study was to find out the expression regulation of CXCL12β by miRNA in inflammation. Materials and Methods: Biopsy samples and blood samples were collected from UC patients and non-IBD controls. mRNA expression was analyzed using microarray and real-time PCR. CXCL12β targeting miRNA were looked by using online target prediction tools. Expression of CXCL12β in blood samples and cell line supernatant was analyzed using ELISA. miRNA target was validated using dual luciferase assay. Results and conclusion: We found miR-200a regulate the expression of CXCL12β in UC. Expression of CXCL12β was increased in cancer susceptible part of colon and expression of its targeting miRNA was decreased in the same part of colon. miR-200a regulate CXCL12β expression in inflammation and may be an important therapeutic target in inflammation associated cancer.

Keywords: inflammation, miRNA, regulation, CXCL12

Procedia PDF Downloads 245

Authors: P. Jost, L. Muckova, M. Matula, J. Pejchal, D. Jun, R. Stetina

Abstract:

Sulfur mustard (bis(2-chlorethyl) sulfide) is highly toxic, chemical warfare agent that has been used in the past in several armed conflicts. Except for the skin, respiratory tract is one of the important routes of exposure. The elucidation and understanding of the mechanism of toxicity of SM have been effort intensive research. The multiple targets character of SM caused cellular damage resulted in activation of many different mechanisms which contribute to cellular response and participate in the final cytopathology effect. In our present work, we compared time-dependent changes in sulfur mustard exposed adult human lung fibroblasts NHLF and lung epithelial alveolar cell line A-549. Cell viability (MTT assay, Calcein-AM assay, and xCELLigence - real-time cell analysis), apoptosis (flow cytometry), mitochondrial membrane potential (Δψm, flow cytometry), reactive oxygen species induction (DC and cell cycle distribution (flow cytometry) were studied. We observed significantly decreased mitochondrial membrane potential and subsequent induction of apoptosis correlating with decreased cellular viability in the sulfur mustard exposed cells. In low concentrations, sulfur mustard-induced S-phase cell cycle arrest, on the other hand, high concentrations, cell cycle phase distribution of sulfur mustard exposed cells resembled cell cycle phase distribution of control group, which implies nonspecific cell cycle inhibition. Epithelial cells A-549 was found as more sensible to sulfur mustard toxicity. Acknowledgements: This work was supported by a long-term organization development plan Medical Aspects of Weapons of Mass Destruction of the Faculty of Military Health Sciences, University of Defence.

Keywords: apoptosis, cell cycle, cytotoxicity, sulfur mustard

Procedia PDF Downloads 164

Authors: Zuhier Altawallbeh

Abstract:

In this paper, an explicit homotopic function is constructed to compute the Hochschild homology of a finite dimensional free k-module V. Because the polynomial algebra is of course fundamental in the computation of the Hochschild homology HH and the cyclic homology CH of commutative algebras, we concentrate our work to compute HH of the polynomial algebra.by providing certain homotopic function.

Keywords: hochschild homology, homotopic function, free and projective modules, free resolution, exterior algebra, symmetric algebra

Procedia PDF Downloads 372

Authors: Shiyin He, Zheng Huang

Abstract:

In this paper, deep learning methods are applied in bio-medical field to detect and count different types of cells in an automatic way instead of manual work in medical practice, specifically in bone marrow examination. The process is mainly composed of two steps, detection and recognition. Mask-Region-Convolutional Neural Networks (Mask-RCNN) was used for detection and image segmentation to extract cells and then Convolutional Neural Networks (CNN), as well as Deep Residual Network (ResNet) was used to classify. Result of cell detection network shows high efficiency to meet application requirements. For the cell recognition network, two networks are compared and the final system is fully applicable.

Keywords: cell detection, cell recognition, deep learning, Mask-RCNN, ResNet

Procedia PDF Downloads 160

Authors: Boudjelthia Radhwane

Abstract:

Self-compacting concrete (SCC) developed in Japan in the late 80s has enabled the construction industry to reduce demand on the resources, improve the work condition and also reduce the impact of environment by elimination of the need for compaction. The shrinkage of concrete is the main cause of cracking in bridge decks. Bridge decks tend to be restrained from shrinkage, and this restraint along with other factors causes the bridge to crack. The characteristics of SCC under restrained shrinkage are important to understand in order to predict the cracking behavior in actual structures. Restrained shrinkage testing is done in accordance to AASHTO testing protocol. The free shrinkage performance and cracking behavior were reported and compared when changing the sand to aggregate ratio and the water to cement ratio. The results of free shrinkage show that when a mix design has higher free shrinkage, it will crack in restrained shrinkage earlier than a mix with lower free shrinkage.

Keywords: concrete mix, cracking behavior, restrained shrinkage, self compacting concrete

Procedia PDF Downloads 353

Authors: Jaehyung Jung, Kiman Kim, Heesang Eum

Abstract:

Solar cell is one of the main technologies to reduce greenhouse gas (GHG). Thereby, accurate estimation of greenhouse gas reduction by solar cell technology is crucial to consider strategic applications of the solar cell. The bottom-up approach using operating data such as operation time and efficiency is one of the methodologies to improve the accuracy of the estimation. In this study, alternative GHG reductions from solar cell technology were estimated by a bottom-up approach to indirect emission source (scope 2) in Korea, 2015. In addition, the scenario-based analysis was conducted to assess the effect of technological change with respect to efficiency improvement and rate of operation. In order to estimate GHG reductions from solar cell activities in operating condition levels, methodologies were derived from 2006 IPCC guidelines for national greenhouse gas inventories and guidelines for local government greenhouse inventories published in Korea, 2016. Indirect emission factors for electricity were obtained from Korea Power Exchange (KPX) in 2011. As a result, the annual alternative GHG reductions were estimated as 21,504 tonCO2eq, and the annual average value was 1,536 tonCO2eq per each solar cell technology. Those results of estimation showed to be 91% levels versus design of capacity. Estimation of individual greenhouse gases (GHGs) showed that the largest gas was carbon dioxide (CO2), of which up to 99% of the total individual greenhouse gases. The annual average GHG reductions from solar cell per year and unit installed capacity (MW) were estimated as 556 tonCO2eq/yr•MW. Scenario analysis of efficiency improvement by 5%, 10%, 15% increased as much as approximately 30, 61, 91%, respectively, and rate of operation as 100% increased 4% of the annual GHG reductions.

Keywords: bottom-up approach, greenhouse gas (GHG), reduction, scenario, solar cell

Procedia PDF Downloads 201

Authors: K. Y. Lin, W. H. Lee, T. W. Cheng, S. W. Huang

Abstract:

Basic Oxygen Furnace (BOF) Slag is a by-product of iron making. It has great engineering properties, such as, high hardness and density, high compressive strength, low abrasion ratio, and can replace natural aggregate for building materials. However, the main problem for BOF slag is expansion, due to it contains free lime or free magnesium. The purpose of this study was to stabilize the BOF slag by using geopolymeric technology, hoping can prevent BOF slag expansion. Geopolymer processes contain a large amount of free silicon. These free silicon can react with free-lime or free magnesium oxide in BOF slag, and thus to form stable compound, therefore inhibit the expansion of the BOF slag. In this study for the successful preparation of geopolymer mortar with BOF slag, and their main properties are analyzed with regard to their use as building materials. Autoclave is used to study the volume stability of these geopolymer mortar. Finally, the compressive strength of geopolymer mortar with BOF slag can be reached 33MPa in 28 days. After autoclave testing, the volume expansion does not exceed 0.2%. Even after the autoclave test, the compressive strength can increase to 35MPa. According to the research results can be proved that using geopolymer technology for stabilizing BOF slag is very effective.

Keywords: BOF slag, autoclave test, geopolymer, swell behavior

Procedia PDF Downloads 111

Authors: Ganesh K. Maurya, Reema Chaudhary, Neha Pandey, Hari S. Misra

Abstract:

PprA, a pleiotropic protein participating in radioresistance, has been reported for its roles in DNA replication initiation, genome segregation, cell division and DNA repair in polyextremophile Deinococcus radiodurans. Interestingly, expression of deinococcal PprA in E. coli suppresses its growth by reducing the number of colony forming units and provides better resistance against γ-radiation than control. We employed different biochemical and cell biology studies using PprA and its DNA binding/polymerization mutants (K133E & W183R) in E. coli. Cells expressing wild type PprA or its K133E mutant showed reduction in the amount of genomic DNA as well as chromosome copy number in comparison to W183R mutant of PprA and control cells, which suggests the role of PprA protein in regulation of DNA replication initiation in E. coli. Further, E. coli cells expressing PprA or its mutants exhibited different impact on cell morphology than control. Expression of PprA or K133E mutant displayed a significant increase in cell length upto 5 folds while W183R mutant showed cell length similar to uninduced control cells. We checked the interaction of deinococcal PprA and its mutants with E. coli DnaA using Bacterial two-hybrid system and co-immunoprecipitation. We observed a functional interaction of EcDnaA with PprA and K133E mutant but not with W183R mutant of PprA. Further, PprA or K133E mutant has suppressed the ATPase activity of EcDnaA but W183R mutant of PprA failed to do so. These observations suggested that PprA protein regulates DNA replication initiation and cell morphology of surrogate E. coli.

Keywords: DNA replication, radioresistance, protein-protein interaction, cell morphology, ATPase activity

Procedia PDF Downloads 36

Authors: Anna Vildová, H. Hendrychová, J. Kubeš, L. Tůmová

Abstract:

The abiotic elicitation is one of the methods for increasing the secondary metabolites production in plant tissue cultures and it seems to be more effective than traditional strategies. This study verified the use of silver nitrate as elicitor to enhance flavonolignans and flavonoid taxifolin production in suspension culture of Sylibum marianum (L.) Gaertn. Silver nitrate in various concentrations (5.887.10-3 mol/L, 5.887.10-4 mol/L, 5.887.10-5 mol/L) was used as elicitor. The content of secondary metabolites in cell suspension cultures was determined by high performance liquid chromatography. The samples were taken after 6, 12, 24, 48, 72 and 168 hours of treatment. The highest content of taxifolin production (2.2 mg.g-1) in cell suspension culture of Silybum marianum (L.) Gaertn. was detected after silver nitrate (5.887.10-4 mol/L) treatment and 72 h application. Flavonolignans such as silybinA, silybin B, silydianin, silychristin, isosilybin A, isosilybin B were not produced by cell suspension culture of S. marianum after elicitor treatment. Our results show that the secondarymetabolites could be released from S. marianum cells into the nutrient medium by changed permeability of cell wall.

Keywords: Silybum marianum (L.) Gaertn., elicitation, silver nitrate, taxifolin

Procedia PDF Downloads 416

Authors: V. Marzal, J. C. Torres, B. Garcia-Camara, Manuel Cano-Garcia, Xabier Quintana, I. Perez Garcilopez, J. M. Sanchez-Pena

Abstract:

At the present time, the use of nanostructures in complex media, like liquid crystals, is widely extended to manipulate their properties, either electrical or optical. In addition, these media can also be used to control the optical properties of the nanoparticles, for instance when they are resonant. In this work, the change on electrical properties of a liquid crystal cell by adding TiO₂ nanoparticles on one of the alignment layers has been analyzed. These nanoparticles, with a diameter of 100 nm and spherical shape, were deposited in one of the substrates (ITO + polyimide) by spin-coating in order to produce a homogeneous layer. These substrates were checked using an optical microscope (objective x100) to avoid potential agglomerates. The liquid crystal cell is then fabricated, using one of these substrates and another without nanoparticles, and filled with E7. The study of the electrical response was done through impedance measurements in a long range of frequencies (3 Hz- 6 MHz) and at ambient temperature. Different nanoparticle concentrations were considered, as well as pure E7 and an empty cell for comparison purposes. Results about the effective dielectric permittivity and conductivity are presented along with models of equivalent electric circuits and its physical interpretation. As a summary, it has been observed the clear influence of the presence of the nanoparticles, strongly modifying the electric response of the device. In particular, a variation of both the effective permittivity and the conductivity of the device have been observed. This result requires a deep analysis of the effect of these nanoparticles on the trapping of free ions in the device, allowing a controlled manipulation and frequency tuning of the electrical response of these devices.

Keywords: alignment layer, electrical behavior, liquid crystal, TiO₂ nanoparticles

Procedia PDF Downloads 181

Authors: Sarah Iaquinta, Christophe Blanquart, Elena Ishow, Sylvain Freour, Frederic Jacquemin, Shahram Khazaie

Abstract:

Nanoparticles have recently emerged as a possible cancer treatment tool. Several formulations have been used to enhance the uptake of these nanoparticles by cancer cells and avoid their immediate clearance when administrated in vivo. Most of the previous studies focus on the investigation of the influence of the mechanical properties of the cell membrane and the particle. However, these studies do not account for the variation of adhesion and tension during the wrapping of the nanoparticle by the membrane. These couplings should be considered since the cell adapts to the interaction with the nanoparticle by, e.g., increasing the number of interactions (consequently leading to an increase of the cell membrane/nanoparticle adhesion) and by reorganizing its cytoskeleton, leading to the releasing of the tension of the cell membrane. The main contribution of this work is the proposal of a novel model for representing the cellular uptake of rigid circular nanoparticles based on an energetic model tailored to take into account the adaptation of the nanoparticle/cell membrane adhesion and of the membrane stress during wrapping. Several coupling models using sigmoidal functions are considered and compared. The study calculations revealed that the results considering constant parameters underestimated the final wrapping degree of the particle by up to 50%.

Keywords: adhesion, cellular adaptation, cellular uptake, mechanical properties, tension

Procedia PDF Downloads 185

Authors: Qiang Xu, Xingxin Wu, Wenjie Guo, Xingqi Wang, Yang Sun, Renxiang Tan

Abstract:

The phosphatase SHP-2 is known to exert regulatory activities on cytokine receptor signaling and the dysregulation of SHP-2 has been implicated in the pathogenesis of a variety of diseases. Here we report several small molecule regulators of SHP-2 for the treatment of T cell-mediated diseases. The new cyclodepsipeptide trichomides A, isolated from the fermentation products of Trichothecium roseum, increased the phosphorylation of SHP-2 in activated T cells, and ameliorated contact dermatitis in mice. The trichomides A’s effects were significantly reversed by using the SHP-2-specific inhibitor PHPS1 or T cell-conditional SHP-2 knockout mice. Another compound is a cerebroside Fusaruside isolated from the endophytic fungus Fusarium sp. IFB-121. Fusaruside also triggered the tyrosine phosphorylation of SHP-2, which provided a possible mean of selectively targeting STAT1 for the treatment of Th1 cell-mediated inflammation and led to the discovery of the non-phosphatase-like function of SHP-2. Namely, the Fusaruside-activated pY-SHP-2 selectively sequestrated the cytosolic STAT1 to prevent its recruitment to IFN-R, which contributed to the improvement of experimental colitis in mice. Blocking the pY-SHP-2-STAT1 interaction, with SHP-2 inhibitor NSC-87877 or using T cells from conditional SHP-2 knockout mice, reversed the effects of fusaruside. Furthermore, the fusaruside’s effect is independent of the phosphatase activity of SHP-2, demonstrating a novel role for SHP-2 in regulating STAT1 signaling and Th1-type immune responses.

Keywords: SHP-2, small molecules, T cell, T cell-mediated diseases

Procedia PDF Downloads 286

Authors: Eric Chabrol, Sidonia B.G. Eckle, Renate de Boer, James McCluskey, Jamie Rossjohn, Mirjam H.M. Heemskerk, Stephanie Gras

Abstract:

αβ and γδ T-cells are disparate T-cell lineages that, via their use of either αβ or γδ T-cell antigen receptors (TCRs) respectively, can respond to distinct antigens. Here we characterise a new population of human T-cells, term δβ T-cells, that express TCRs comprising a TCR-δ variable gene fused to a Joining-α/Constant-α domain, paired with an array of TCR-β chains. We characterised the cellular, functional, biophysical and structural characteristic feature of this new T-cells population that reveal some new insight into TCR diversity. We provide molecular bases of how δβ T-cells can recognise viral peptide presented by Human Leukocyte Antigen (HLA) molecule. Our findings highlight how components from αβ and γδTCR gene loci can recombine to confer antigen specificity thus expanding our understanding of T-cell biology and TCR diversity.

Keywords: new delta-beta TCR, HLA, viral peptide, structural immunology

Procedia PDF Downloads 400