Search results for: beta-barrel membrane proteins
939 Targeted Delivery of Novel Copper-Based Nanoparticles for Advance Cancer Therapeutics
Authors: Arindam Pramanik, Parimal Karmakar
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We have explored the synergistic anti-cancer activity of copper ion and acetylacetone complex containing 1,3 diketone group (like curcumin) in metallorganic compound “Copper acetylacetonate” (CuAA). The cytotoxicity mechanism of CuAA complex was evaluated on various cancer cell lines in vitro. Among these, reactive oxygen species (ROS), glutathione level (GSH) in the cell was found to increase. Further mitochondrial membrane damage was observed. The fate of cell death was found to be induced by apoptosis. For application purpose, we have developed a novel biodegradable, non-toxic polymer-based nanoparticle which has hydrophobically modified core for loading of the CuAA. Folic acid is conjugated on the surface of the polymer (chitosan) nanoparticle for targeting to cancer cells for minimizing toxicity to normal cells in-vivo. Thus, this novel drug CuAA has an efficient anticancer activity which has been targeted specifically to cancer cells through polymer nanoparticle.Keywords: anticancer, apoptosis, copper nanoparticle, targeted drug delivery
Procedia PDF Downloads 473938 Non-Steroidal Microtubule Disrupting Analogues Induce Programmed Cell Death in Breast and Lung Cancer Cell Lines
Authors: Marcel Verwey, Anna M. Joubert, Elsie M. Nolte, Wolfgang Dohle, Barry V. L. Potter, Anne E. Theron
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A tetrahydroisoquinolinone (THIQ) core can be used to mimic the A,B-ring of colchicine site-binding microtubule disruptors such as 2-methoxyestradiol in the design of anti-cancer agents. Steroidomimeric microtubule disruptors were synthesized by introducing C'2 and C'3 of the steroidal A-ring to C'6 and C'7 of the THIQ core and by introducing a decorated hydrogen bond acceptor motif projecting from the steroidal D-ring to N'2. For this in vitro study, four non-steroidal THIQ-based analogues were investigated and comparative studies were done between the non-sulphamoylated compound STX 3450 and the sulphamoylated compounds STX 2895, STX 3329 and STX 3451. The objective of this study was to investigate the modes of cell death induced by these four THIQ-based analogues in A549 lung carcinoma epithelial cells and metastatic breast adenocarcinoma MDA-MB-231 cells. Cytotoxicity studies to determine the half maximal growth inhibitory concentrations were done using spectrophotometric quantification via crystal violet staining and lactate dehydrogenase (LDH) assays. Microtubule integrity and morphologic changes of exposed cells were investigated using polarization-optical transmitted light differential interference contrast microscopy, transmission electron microscopy and confocal microscopy. Flow cytometric quantification was used to determine apoptosis induction and the effect that THIQ-based analogues have on cell cycle progression. Signal transduction pathways were elucidated by quantification of the mitochondrial membrane integrity, cytochrome c release and caspase 3, -6 and -8 activation. Induction of autophagic cell death by the THIQ-based analogues was investigated by morphological assessment of fluorescent monodansylcadaverine (MDC) staining of acidic vacuoles and by quantifying aggresome formation via flow cytometry. Results revealed that these non-steroidal microtubule disrupting analogues inhibited 50% of cell growth at nanomolar concentrations. Immunofluorescence microscopy indicated microtubule depolarization and the resultant mitotic arrest was further confirmed through cell cycle analysis. Apoptosis induction via the intrinsic pathway was observed due to depolarization of the mitochondrial membrane, induction of cytochrome c release as well as, caspase 3 activation. Potential involvement of programmed cell death type II was observed due to the presence of acidic vacuoles and aggresome formation. Necrotic cell death did not contribute significantly, indicated by stable LDH levels. This in vitro study revealed the induction of the intrinsic apoptotic pathway as well as possible involvement of autophagy after exposure to these THIQ-based analogues in both MDA-MB-231- and A549 cells. Further investigation of this series of anticancer drugs still needs to be conducted to elucidate the temporal, mechanistic and functional crosstalk mechanisms between the two observed programmed cell deaths pathways.Keywords: apoptosis, autophagy, cancer, microtubule disruptor
Procedia PDF Downloads 239937 Stability Enhancement of Supported Ionic Liquid Membranes Using Ion Gels for Gas Separation
Authors: Y. H. Hwang, J. Won, Y. S. Kang
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Supported ionic liquid membranes (SILMs) have attracted due to the negligible vapor pressure of ionic liquids (ILs) as well as the high gas selectivity for specific gases such as CO2 or olefin. 1-ethyl-3-methylimidazolium tricyanomethanide ([EMIM][TCM]), 1-butyl-3-methylimidazolium tricyanomethanide ([BMIM][TCM]), show high CO2 solubility, CO2 absorption, rapid CO2 absorption rate and negligible vapor pressure, SILMs using these ILs have been good candidates as CO2 separation membranes. However, SILM has to be operated at a low differential pressure to prevent the solvent from being expelled from the pores of supported membranes. In this paper, we improve the mechanical strength by forming ion gels which provide the stability while it retains the diffusion properties of the liquid stage which affects the gas separation properties. The ion gel was created by the addition of tri-block copolymer, poly(styrene-ethylene oxide-b-styrene) in RTIL. SILM using five different RTILs, are investigated with and without ion gels. The gas permeance were measured and the gas performance with and without the SEOS were investigated.Keywords: ion gel, ionic liquid, membrane, nanostructure
Procedia PDF Downloads 305936 Urea Amperometric Biosensor Based on Entrapment Immobilization of Urease onto a Nanostructured Polypyrrol and Multi-Walled Carbon Nanotube
Authors: Hamide Amani, Afshin FarahBakhsh, Iman Farahbakhsh
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In this paper, an amprometric biosensor based on surface modified polypyrrole (PPy) has been developed for the quantitative estimation of urea in aqueous solutions. The incorporation of urease (Urs) into a bipolymeric substrate consisting of PPy was performed by entrapment to the polymeric matrix, PPy acts as amperometric transducer in these biosensors. To increase the membrane conductivity, multi-walled carbon nanotubes (MWCNT) were added to the PPy solution. The entrapped MWCNT in PPy film and the bipolymer layers were prepared for construction of Pt/PPy/MWCNT/Urs. Two different configurations of working electrodes were evaluated to investigate the potential use of the modified membranes in biosensors. The evaluation of two different configurations of working electrodes suggested that the second configuration, which was composed of an electrode-mediator-(pyrrole and multi-walled carbon nanotube) structure and enzyme, is the best candidate for biosensor applications.Keywords: urea biosensor, polypyrrole, multi-walled carbon nanotube, urease
Procedia PDF Downloads 312935 Physical Characteristics of Cookies Enriched with Microencapsulated Cherry Pomace Extract
Authors: Jovana Petrović, Ivana Lončarević, Vesna Tumbas Šaponjac, Biljana Pajin, Danica Zarić
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Pomace, a by-product from fruit processing industry is the potential source of valuable bioactive. Cookies are popular, ready to eat and low price foods; therefore, enrichment of these products is of great importance. In this work, bioactive compounds extracted from cherry pomace, encapsulated in soy and whey proteins, have been incorporated in cookies, replacing 10 (SP10 and WP10) and 15% of wheat flour (SP15 and WP15). Cookie geometry (diameter (D), thickness (T) and spread ratio (D/T)), cookie weight, cookie hardness and cookie surface colour were measured. Sensory characteristics are also examined. The results show that encapsulated cherry pomace bioactives have positively influenced the cookie mass. Diameter, redness (a* value) and cookie hardness increased. Sensory evaluation of cookies, revealed that up to 15% substitution of wheat flour with WP encapsulate produced acceptable cookies similar to the control (100% wheat flour) cookies.Keywords: cherry pomace, polyphenols, microencapsulation, cookies, physical characteristics
Procedia PDF Downloads 460934 In-Silico Investigation of Phytochemicals from Ocimum Sanctum as Plausible Antiviral Agent in COVID-19
Authors: Dileep Kumar, Janhavi Ramchandra Rao Kumar, Rao
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COVID-19 has ravaged the globe, and it is spreading its Spectre day by day. In the absence of established drugs, this disease has created havoc. Some of the infected persons are symptomatic or asymptomatic. The respiratory system, cardiac system, digestive system, etc. in human beings are affected by this virus. In our present investigation, we have undertaken a study of the Indian Ayurvedic herb, Ocimum sanctum against SARS-CoV-2 using molecular docking and dynamics studies. The docking analysis was performed on the Glide module of Schrödinger suite on two different proteins from SARS-CoV-2 viz. NSP15 Endoribonuclease and spike receptor-binding domain. MM-GBSA based binding free energy calculations also suggest the most favorable binding affinities of carvacrol, β elemene, and β caryophyllene with binding energies of −61.61, 58.23, and −54.19 Kcal/mol respectively with spike receptor-binding domain and NSP15 Endoribonuclease. It rekindles our hope for the design and development of new drug candidates for the treatment of COVID19.Keywords: molecular docking, COVID-19, ocimum sanctum, binding energy
Procedia PDF Downloads 175933 Surface Adjustments for Endothelialization of Decellularized Porcine Pericardium
Authors: M. Markova, E. Filova, O. Kaplan, R. Matejka, L. Bacakova
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The porcine pericardium is used as a material for cardiac and aortic valves substitutes. Current biological aortic heart valve prosthesis have a limited lifetime period because they undergo degeneration. In order to make them more biocompatible and prolong their lifetime it is necessary to reseed the decellularized prostheses with endothelial cells and with valve interstitial cells. The endothelialization of the prosthesis-surface may be supported by suitable chemical surface modification of the prosthesis. The aim of this study is to prepare bioactive fibrin layers which would both support endothelialization of porcine pericardium and enhance differentiation and maturation of the endothelial cells seeded. As a material for surface adjustments we used layers of fibrin with/without heparin and some of them with adsorbed or chemically bound FGF2, VEGF or their combination. Fibrin assemblies were prepared in 24-well cell culture plate and were seeded with HSVEC (Human Saphenous Vein Endothelial Cells) at a density of 20,000 cells per well in EGM-2 medium with 0.5% FS and without heparin, without FGF2 and without VEGF; medium was supplemented with aprotinin (200 U/mL). As a control, surface polystyrene (PS) was used. Fibrin was also used as homogeneous impregnation of the decellularized porcine pericardium throughout the scaffolds. Morphology, density, and viability of the seeded endothelial cells were observed from micrographs after staining the samples by LIVE/DEAD cytotoxicity/viability assay kit on the days 1, 3, and 7. Endothelial cells were immunocytochemically stained for proteins involved in cell adhesion, i.e. alphaV integrin, vinculin, and VE-cadherin, markers of endothelial cells differentiation and maturation, i.e. von Willebrand factor and CD31, and for extracellular matrix proteins typically produced by endothelial cells, i.e. type IV collagen and laminin. The staining intensities were subsequently quantified using a software. HSVEC cells grew on each of the prepared surfaces better than on control surface. They reached confluency. The highest cell densities were obtained on the surface of fibrin with heparin and both grow factors used together. Intensity of alphaV integrins staining was highest on samples with remained fibrin layer, i.e. on layers with lower cell densities, i.e. on fibrin without heparin. Vinculin staining was apparent, but was rather diffuse, on fibrin with both FGF2 and VEGF and on control PS. Endothelial cells on all samples were positively stained for von Willebrand factor and CD31. VE-cadherin receptors clusters were best developed on fibrin with heparin and growth factors. Significantly stronger staining of type IV collagen was observed on fibrin with heparin and both growth factors. Endothelial cells on all samples produced laminin-1. Decellularized pericardium was homogeneously filled with fibrin structures. These fibrin-modified pericardium samples will be further seeded with cells and cultured in a bioreactor. Fibrin layers with/without heparin and with adsorbed or chemically bound FGF2, VEGF or their combination are good surfaces for endothelialization of cardiovascular prostheses or porcine pericardium based heart valves. Supported by the Ministry of Health, grants No15-29153A and 15-32497A, and the Grant Agency of the Czech Republic, project No. P108/12/G108.Keywords: aortic valves prosthesis, FGF2, heparin, HSVEC cells, VEGF
Procedia PDF Downloads 255932 Energy Self-Sufficiency Through Smart Micro-Grids and Decentralised Sector-Coupling
Authors: C. Trapp, A. Vijay, M. Khorasani
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Decentralised micro-grids with sector coupling can combat the spatial and temporal intermittence of renewable energy by combining power, transportation and infrastructure sectors. Intelligent energy conversion concepts such as electrolysers, hydrogen engines and fuel cells combined with energy storage using intelligent batteries and hydrogen storage form the back-bone of such a system. This paper describes a micro-grid based on Photo-Voltaic cells, battery storage, innovative modular and scalable Anion Exchange Membrane (AEM) electrolyzer with an efficiency of up to 73%, high-pressure hydrogen storage as well as cutting-edge combustion-engine based Combined Heat and Power (CHP) plant with more than 85% efficiency at the university campus to address the challenges of decarbonization whilst eliminating the necessity for expensive high-voltage infrastructure.Keywords: sector coupling, micro-grids, energy self-sufficiency, decarbonization, AEM electrolysis, hydrogen CHP
Procedia PDF Downloads 168931 The Role of P2X7 Cytoplasmic Anchor in Inflammation
Authors: Federico Cevoli
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Purinergic P2X7 receptors (P2X7R) are ligand-gated non-selective cation channels involved in several physiological and pathological processes. They are particularly promising pharmacological targets as they are present in an increasing number of different cells types. P2X7R activation is triggered following elevated concentrations of extracellular ATP, similarly to those observed in tissues injury, chronic inflammation and T-cell activation, as well as in the scrambling of phospholipids leading to membrane blebbing and apoptosis. Another hallmark of P2X7 is cell permeabilization, commonly known as “macropore” formation allowing the passage of nanometer-sized molecules up to 900Da. Recently, full-length P2X7 Cryo-EM structures revealed unique functional sites, including two cytoplasmic domains - the cytoplasmic "anchor" and "ballast". To date, the molecular units/complex by which P2X7R exerts its pathophysiological functions are unknown. Using custom-made cell-penetrating HIV-1 TAT peptides, we show for the first-time potential implications of P2X7 cytoplasmic anchor in the regulation of caspase3/7 activation as well as TNFα regulation.Keywords: P2X7R, immunology, TAT-peptide, cell death
Procedia PDF Downloads 119930 Genetic Polymorphism and Insilico Study Epitope Block 2 MSP1 Gene of Plasmodium falciparum Isolate Endemic Jayapura
Authors: Arsyam Mawardi, Sony Suhandono, Azzania Fibriani, Fifi Fitriyah Masduki
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Malaria is an infectious disease caused by Plasmodium sp. This disease has a high prevalence in Indonesia, especially in Jayapura. The vaccine that is currently being developed has not been effective in overcoming malaria. This is due to the high polymorphism in the Plasmodium genome especially in areas that encode Plasmodium surface proteins. Merozoite Surface Protein 1 (MSP1) Plasmodium falciparum is a surface protein that plays a role in the invasion process in human erythrocytes through the interaction of Glycophorin A protein receptors and sialic acid in erythrocytes with Reticulocyte Binding Proteins (RBP) and Duffy Adhesion Protein (DAP) ligands in merozoites. MSP1 can be targeted to be a specific antigen and predicted epitope area which will be used for the development of diagnostic and malaria vaccine therapy. MSP1 consists of 17 blocks, each block is dimorphic, and has been marked as the K1 and MAD20 alleles. Exceptions only in block 2, because it has 3 alleles, among others K1, MAD20 and RO33. These polymorphisms cause allelic variations and implicate the severity of patients infected P. falciparum. In addition, polymorphism of MSP1 in Jayapura isolates has not been reported so it is interesting to be further identified and projected as a specific antigen. Therefore, in this study, we analyzed the allele polymorphism as well as detected the MSP1 epitope antigen candidate on block 2 P. falciparum. Clinical samples of selected malaria patients followed the consecutive sampling method, examining malaria parasites with blood preparations on glass objects observed through a microscope. Plasmodium DNA was isolated from the blood of malarial positive patients. The block 2 MSP1 gene was amplified using PCR method and cloned using the pGEM-T easy vector then transformed to TOP'10 E.coli. Positive colonies selection was performed with blue-white screening. The existence of target DNA was confirmed by PCR colonies and DNA sequencing methods. Furthermore, DNA sequence analysis was done through alignment and formation of a phylogenetic tree using MEGA 6 software and insilico analysis using IEDB software to predict epitope candidate for P. falciparum. A total of 15 patient samples have been isolated from Plasmodium DNA. PCR amplification results show the target gene size about ± 1049 bp. The results of MSP1 nucleotide alignment analysis reveal that block 2 MSP1 genes derived from the sample of malarial patients were distributed in four different allele family groups, K1 (7), MAD20 (1), RO33 (0) and MSP1_Jayapura (10) alleles. The most commonly appears of the detected allele is MSP1_Jayapura single allele. There was no significant association between sex variables, age, the density of parasitemia and alel variation (Mann Whitney, U > 0.05), while symptomatic signs have a significant difference as a trigger of detectable allele variation (U < 0.05). In this research, insilico study shows that there is a new epitope antigen candidate from the MSP1_Jayapura allele and it is predicted to be recognized by B cells with 17 amino acid lengths in the amino acid sequence 187 to 203.Keywords: epitope candidate, insilico analysis, MSP1 P. falciparum, polymorphism
Procedia PDF Downloads 172929 Improved Operating Strategies for the Optimization of Proton Exchange Membrane Fuel Cell System Performance
Authors: Guillaume Soubeyran, Fabrice Micoud, Benoit Morin, Jean-Philippe Poirot-Crouvezier, Magali Reytier
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Proton Exchange Membrane Fuel Cell (PEMFC) technology is considered as a solution for the reduction of CO2 emissions. However, this technology still meets several challenges for high-scale industrialization. In this context, the increase of durability remains a critical aspect for competitiveness of this technology. Fortunately, performance degradations in nominal operating conditions is partially reversible, meaning that if specific conditions are applied, a partial recovery of fuel cell performance can be achieved, while irreversible degradations can only be mitigated. Thus, it is worth studying the optimal conditions to rejuvenate these reversible degradations and assessing the long-term impact of such procedures on the performance of the cell. Reversible degradations consist mainly of anode Pt active sites poisoning by carbon monoxide at the anode, heterogeneities in water management during use, and oxidation/deactivation of Pt active sites at the cathode. The latter is identified as a major source of reversible performance loss caused by the presence oxygen, high temperature and high cathode potential that favor platinum oxidation, especially in high efficiency operating points. Hence, we studied here a recovery procedure aiming at reducing the platinum oxides by decreasing cathode potential during operation. Indeed, the application of short air starvation phase leads to a drop of cathode potential. Cell performances are temporarily increased afterwards. Nevertheless, local temperature and current heterogeneities within the cells are favored and shall be minimized. The consumption of fuel during the recovery phase shall also be considered to evaluate the global efficiency. Consequently, the purpose of this work is to find an optimal compromise between the recovery of reversible degradations by air starvation, the increase of global cell efficiency and the mitigation of irreversible degradations effects. Different operating parameters have first been studied such as cell voltage, temperature and humidity in single cell set-up. Considering the global PEMFC system efficiency, tests showed that reducing duration of recovery phase and reducing cell voltage was the key to ensure an efficient recovery. Recovery phase frequency was a major factor as well. A specific method was established to find the optimal frequency depending on the duration and voltage of the recovery phase. Then, long-term degradations have also been studied by applying FC-DLC cycles based on NEDC cycles on a 4-cell short stack by alternating test sequences with and without recovery phases. Depending on recovery phase timing, cell efficiency during the cycle was increased up to 2% thanks to a mean voltage increase of 10 mV during test sequences with recovery phases. However, cyclic voltammetry tests results suggest that the implementation of recovery phases causes an acceleration of the decrease of platinum active areas that could be due to the high potential variations applied to the cathode electrode during operation.Keywords: durability, PEMFC, recovery procedure, reversible degradation
Procedia PDF Downloads 122928 Preliminary Study on Using of Thermal Energy from Effluent Water for the SBR Process of RO
Authors: Gyeong-Sung Kim, In-soo Ahn, Yong Cho
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SBR (Sequencing Batch Reactor) process is usually applied to membrane water treatment plants to treat its concentrated wastewater. The role of SBR process is to remove COD (Chemical Oxygen Demand) and NH3 from wastewater before discharging it outside of the water treatment plant using microorganism. Microorganism’s nitrification capability is influenced by water temperature because the nitrification rate of the concentrated wastewater becomes ‘zero’ as water temperature approach 0℃. Heating system is necessary to operate SBR in winter season even though the operating cost increase sharply. The operating cost of SBR at ‘D’ RO water treatment plant in Korea was 51.8 times higher in winter (October to March) compare to summer (April to September) season in 2014. Otherwise the effluent water temperature maintained around 8℃ constantly in winter. This study focuses on application heat pump system to recover the thermal energy from the effluent water of ‘D’ RO plant so that the operating cost will be reduced.Keywords: water treatment, water thermal energy, energy saving, RO, SBR
Procedia PDF Downloads 508927 A Rare Atypical Presentation of Iichthyosis Follicularis, Alopecia, and Photophobia Syndrome
Authors: D. R. Apoorva
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Ichthyosis follicularis, alopecia, and photophobia (IFAP) syndrome is a rare oculocutaneous disorder of genetic origin. This disorder results from mutations in the membrane-bound transcription factor protease site, two genes that impair cholesterol homeostasis, and the ability to cope with endoplasmic reticulum stress. We report a rare case of IFAP syndrome with an atypical presentation, and it was interesting to note that the child had patchy non-scarring alopecia over the scalp along with unilateral madarosis. To our best knowledge, this unique presentation has not been described earlier. The child presented with photophobia and unilateral ptosis. The child also had short stature and intellectual disability. Skin histopathology was nonspecific and consisted of dilated hair follicles with keratin plugs extending above the skin surface. This rare oculocutaneous disorder requires proper documentation so that identification of its variants may be possible in the future. Early recognition of atypical presentations can help in preventing cardiovascular complications, which remain the major cause of death.Keywords: alopecia, photophobia, ichthyosis follicularis, IFAP syndrome
Procedia PDF Downloads 68926 Modelling and Simulation of Milk Fouling
Authors: Harche Rima, Laoufi Nadia Aicha
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This work focuses on the study and modeling of the fouling phenomenon in a vertical pipe. In the first step, milk is one of the fluids obeying the phenomenon of fouling because of the denaturation of these proteins, especially lactoglobulin, which is the active element of milk, and to facilitate its use, we chose to study milk as a fouling fluid. In another step, we consider the test section of our installation as a tubular-type heat exchanger that works against the current and in a closed circuit. A simple mathematical model of Kern & Seaton, based on the kinetics of the fouling resistance, was used to evaluate the influence of the operating parameters (fluid flow velocity and exchange wall temperature) on the fouling resistance. The influence of the variation of the fouling resistance with the operating conditions on the efficiency of the heat exchanger and the importance of the dirty state exchange coefficient as an exchange quality control parameter were discussed and examined. On the other hand, an electronic scanning microscope analysis was performed on the milk deposit in order to obtain its actual image and composition, which allowed us to calculate the thickness of this deposit.Keywords: fouling, milk, tubular heat exchanger, fouling resistance
Procedia PDF Downloads 35925 The Impact of Steel Connections on the Fire Resistance of Composite Buildings
Authors: Shuyuan Lin, Zhaohui Huang, Mizi Fan
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In the majority of previous research into modelling large scale composite floor subjected to fire, the beam-to-column and beam-to-beam connections were assumed to behave either as pinned or rigid for simplicity, and the vertical shear and axial tension failures of the connection were not taken into account. We have recently developed robust two-noded connection models for modeling endplate and partial endplate steel connections under fire conditions. The main objective of this research is to systematically investigate the impact of the connections of protected beams, on the tensile membrane actions of supported floor slabs in which the failures of the connections, such as, axial tension, vertical shear and bending are accounted for. The models developed have very good numerical stability under a static solver condition, and can be used for large scale modelling of composite buildings in fire.Keywords: fire, steel structure, component-based model, beam-to-column connections
Procedia PDF Downloads 442924 Designing Next Generation Platforms for Recombinant Protein Production by Genome Engineering of Escherichia coli
Authors: Priyanka Jain, Ashish K. Sharma, Esha Shukla, K. J. Mukherjee
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We propose a paradigm shift in our approach to design improved platforms for recombinant protein production, by addressing system level issues rather than the individual steps associated with recombinant protein synthesis like transcription, translation, etc. We demonstrate that by controlling and modulating the cellular stress response (CSR), which is responsible for feedback control of protein synthesis, we can generate hyper-producing strains. We did transcriptomic profiling of post-induction cultures, expressing different types of protein, to analyze the nature of this cellular stress response. We found significant down-regulation of substrate utilization, translation, and energy metabolism genes due to generation CSR inside the host cell. However, transcription profiling has also shown that many genes are up-regulated post induction and their role in modulating the CSR is unclear. We hypothesized that these up-regulated genes trigger signaling pathways, generating the CSR and concomitantly reduce the recombinant protein yield. To test this hypothesis, we knocked out the up-regulated genes, which did not have any downstream regulatees, and analyzed their impact on cellular health and recombinant protein expression. Two model proteins i.e., GFP and L-Asparaginase were chosen for this analysis. We observed a significant improvement in expression levels, with some knock-outs showing more than 7-fold higher expression compared to control. The 10 best single knock-outs were chosen to make 45 combinations of all possible double knock-outs. A further increase in expression was observed in some of these double knock- outs with GFP levels being highest in a double knock-out ΔyhbC + ΔelaA. However, for L-Asparaginase which is a secretory protein, the best results were obtained using a combination of ΔelaA+ΔcysW knock-outs. We then tested all the knock outs for their ability to enhance the expression of a 'difficult-to-express' protein. The Rubella virus E1 protein was chosen and tagged with sfGFP at the C-terminal using a linker peptide for easy online monitoring of expression of this fusion protein. Interestingly, the highest increase in Rubella-sGFP levels was obtained in the same double knock-out ΔelaA + ΔcysW (5.6 fold increase in expression yield compared to the control) which gave the highest expression for L-Asparaginase. However, for sfGFP alone, the ΔyhbC+ΔmarR knock-out gave the highest level of expression. These results indicate that there is a fair degree of commonality in the nature of the CSR generated by the induction of different proteins. Transcriptomic profiling of the double knock out showed that many genes associated with the translational machinery and energy biosynthesis did not get down-regulated post induction, unlike the control where these genes were significantly down-regulated. This confirmed our hypothesis of these genes playing an important role in the generation of the CSR and allowed us to design a strategy for making better expression hosts by simply knocking out key genes. This strategy is radically superior to the previous approach of individually up-regulating critical genes since it blocks the mounting of the CSR thus preventing the down-regulation of a very large number of genes responsible for sustaining the flux through the recombinant protein production pathway.Keywords: cellular stress response, GFP, knock-outs, up-regulated genes
Procedia PDF Downloads 219923 Antibacterial Activity of Libyan Seaweed Extracts
Authors: Salmin K. Alshalmani, Nada H. Zobi, Ismaeel H. Bozakouk
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Marine organisms are potentially prolific sources of highly bio active secondary metabolites that might represent useful leads in the development of new pharmaceutical agents. The Libyan marine biodiversity including macroalgae remains partially unexplored in term of their potential bio activities. The phytochemical analysis of the alcoholic extracts of some commonly occurring seaweed Cystoseira compressa, enteromorpha intestinals, corallina, and Ulva lactuca and their evaluated for antibacterial activity by well diffusion assay were studied. Four different solvents namely water, ethanol 99 %, methanol 99 %, and methylated spirit 95 % were used for extraction. The phytochemical analysis revealed the presence of carbohydrates, steroids, tannin & phenols, saponins, proteins, and glycosides. The extracts were subjected for study of antibacterial activity. The zone of inhibition ranged between 8 to 16 mm in aqueous extract and up to 16 mm in methanol extract. The maximum activity (16 mm) was recorded from methanol extract of Ulva lactuca against Staphylococcus aureus and, minimum activity (8mm) recorded by Cystoseira compressa against S. aureus.Keywords: macroalgae, phytochemicals, antibacterial activity, methanolic extract
Procedia PDF Downloads 457922 Construction and Cross-Linking of Polyelectrolyte Multilayers Based on Polysaccharides as Antifouling Coatings
Authors: Wenfa Yu, Thuva Gnanasampanthan, John Finlay, Jessica Clarke, Charlotte Anderson, Tony Clare, Axel Rosenhahn
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Marine biofouling is a worldwide problem at vast economic and ecological costs. Historically it was combated with toxic coatings such as tributyltin. As those coatings being banned nowadays, finding environmental friendly antifouling solution has become an urgent topic. In this study antifouling coatings consisted of natural occurring polysaccharides hyaluronic acid (HA), alginic acid (AA), chitosan (Ch) and polyelectrolyte polyethylenimine (PEI) are constructed into polyelectrolyte multilayers (PEMs) in a Layer-by-Layer (LbL) method. LbL PEM construction is a straightforward way to assemble biomacromolecular coatings on surfaces. Advantages about PEM include ease of handling, highly diverse PEM composition, precise control over the thickness and so on. PEMs have been widely employed in medical application and there are numerous studies regarding their protein adsorption, elasticity and cell adhesive properties. With the adjustment of coating composition, termination layer charge, coating morphology and cross-linking method, it is possible to prepare low marine biofouling coatings with PEMs. In this study, using spin coating technology, PEM construction was achieved at smooth multilayers with roughness as low as 2nm rms and highly reproducible thickness around 50nm. To obtain stability in sea water, the multilayers were covalently cross-linked either thermally or chemically. The cross-linking method affected surface energy, which was reflected in water contact angle, thermal cross-linking led to hydrophobic surfaces and chemical cross-linking generated hydrophilic surfaces. The coatings were then evaluated regarding its protein resistance and biological species resistance. While the hydrophobic thermally cross-linked PEM had low resistance towards proteins, the resistance of chemically cross-linked PEM strongly depended on the PEM termination layer and the charge of the protein, opposite charge caused high adsorption and same charge low adsorption, indicating electrostatic interaction plays a crucial role in the protein adsorption processes. Ulva linza was chosen as the biological species for antifouling performance evaluation. Despite of the poor resistance towards protein adsorption, thermally cross-linked PEM showed good resistance against Ulva spores settlement, the chemically cross-linked multilayers showed poor resistance regardless of the termination layer. Marine species adhesion is a complex process, although it involves proteins as bioadhesives, protein resistance its own is not a fully indicator for its antifouling performance. The species will pre select the surface, responding to cues like surface energy, chemistry, or charge and so on. Thus making it difficult for one single factors to determine its antifouling performance. Preparing PEM coating is a comprehensive work involving choosing polyelectrolyte combination, determining termination layer and the method for cross-linking. These decisions will affect PEM properties such as surface energy, charge, which is crucial, since biofouling is a process responding to surface properties in a highly sensitive and dynamic way.Keywords: hyaluronic acid, polyelectrolyte multilayers, protein resistance, Ulva linza zoospores
Procedia PDF Downloads 156921 PARP1 Links Transcription of a Subset of RBL2-Dependent Genes with Cell Cycle Progression
Authors: Ewelina Wisnik, Zsolt Regdon, Kinga Chmielewska, Laszlo Virag, Agnieszka Robaszkiewicz
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Apart from protecting genome, PARP1 has been documented to regulate many intracellular processes inter alia gene transcription by physically interacting with chromatin bound proteins and by their ADP-ribosylation. Our recent findings indicate that expression of PARP1 decreases during the differentiation of human CD34+ hematopoietic stem cells to monocytes as a consequence of differentiation-associated cell growth arrest and formation of E2F4-RBL2-HDAC1-SWI/SNF repressive complex at the promoter of this gene. Since the RBL2 complexes repress genes in a E2F-dependent manner and are widespread in the genome in G0 arrested cells, we asked (a) if RBL2 directly contributes to defining monocyte phenotype and function by targeting gene promoters and (b) if RBL2 controls gene transcription indirectly by repressing PARP1. For identification of genes controlled by RBL2 and/or PARP1,we used primer libraries for surface receptors and TLR signaling mediators, genes were silenced by siRNA or shRNA, analysis of gene promoter occupation by selected proteins was carried out by ChIP-qPCR, while statistical analysis in GraphPad Prism 5 and STATISTICA, ChIP-Seq data were analysed in Galaxy 2.5.0.0. On the list of 28 genes regulated by RBL2, we identified only four solely repressed by RBL2-E2F4-HDAC1-BRM complex. Surprisingly, 24 out of 28 emerged genes controlled by RBL2 were co-regulated by PARP1 in six different manners. In one mode of RBL2/PARP1 co-operation, represented by MAP2K6 and MAPK3, PARP1 was found to associate with gene promoters upon RBL2 silencing, which was previously shown to restore PARP1 expression in monocytes. PARP1 effect on gene transcription was observed only in the presence of active EP300, which acetylated gene promoters and activated transcription. Further analysis revealed that PARP1 binding to MA2K6 and MAPK3 promoters enabled recruitment of EP300 in monocytes, while in proliferating cancer cell lines, which actively transcribe PARP1, this protein maintained EP300 at the promoters of MA2K6 and MAPK3. Genome-wide analysis revealed a similar distribution of PARP1 and EP300 around transcription start sites and the co-occupancy of some gene promoters by PARP1 and EP300 in cancer cells. Here, we described a new RBL2/PARP1/EP300 axis which controls gene transcription regardless of the cell type. In this model cell, cycle-dependent transcription of PARP1 regulates expression of some genes repressed by RBL2 upon cell cycle limitation. Thus, RBL2 may indirectly regulate transcription of some genes by controlling the expression of EP300-recruiting PARP1. Acknowledgement: This work was financed by Polish National Science Centre grants nr DEC-2013/11/D/NZ2/00033 and DEC-2015/19/N/NZ2/01735. L.V. is funded by the National Research, Development and Innovation Office grants GINOP-2.3.2-15-2016-00020 TUMORDNS, GINOP-2.3.2-15-2016-00048-STAYALIVE and OTKA K112336. AR is supported by Polish Ministry of Science and Higher Education 776/STYP/11/2016.Keywords: retinoblastoma transcriptional co-repressor like 2 (RBL2), poly(ADP-ribose) polymerase 1 (PARP1), E1A binding protein p300 (EP300), monocytes
Procedia PDF Downloads 197920 Cloning and Analysis of Nile Tilapia Toll-like receptors Type-3 mRNA
Authors: Abdelazeem Algammal, Reham Abouelmaatti, Xiaokun Li, Jisheng Ma, Eman Abdelnaby, Wael Elfeil
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Toll-like receptors (TLRs) are the best understood of the innate immune receptors that detect infections in vertebrates. However, the fish TLRs also exhibit very distinct features and a large diversity, which is likely derived from their diverse evolutionary history and the distinct environments that they occupy. Little is known about the fish immune system structure. Our work was aimed to identify and clone the Nile tilapiaTLR-3 as a model of freshwater fish species; we cloned the full-length cDNA sequence of Nile tilapia (Oreochromis niloticus) TLR-3 and according to our knowledge, it is the first report illustrating tilapia TLR-3. The complete cDNA sequence of Nile tilapia TLR-3 was 2736 pair base and it encodes a polypeptide of 912 amino acids. Analysis of the deduced amino acid sequence indicated that Nile tilapia TLR-3 has typical structural features and main components of proteins belonging to the TLR family. Our results illustrate a complete and functional Nile tilapia TLR-3 and it is considered an ortholog of the other vertebrate’s receptor.Keywords: Nile tilapia, TLR-3, cloning, gene expression
Procedia PDF Downloads 142919 Soil Arching Effect in Columnar Embankments: A Numerical Study
Authors: Riya Roy, Anjana Bhasi
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Column-supported embankments provide a practical and efficient solution for construction on soft soil due to the low cost and short construction times. In the recent years, geosynthetic have been used in combination with column systems to support embankments. The load transfer mechanism in these systems is a combination of soil arching effect, which occurs between columns and membrane effect of the geosynthetic. This paper aims at the study of soil arching effect on columnar embankments using finite element software, ABAQUS. An axisymmetric finite element model is generated and using this model, parametric studies are carried out. Thus the effects of various factors such as height of embankment fill, elastic modulus of pile and tensile stiffness of geosynthetic, on soil arching have been studied. The development of negative skin friction along the pile-soil interface have also been studied and the results obtained from this study are compared with the current design methods.Keywords: ABAQUS, geosynthetic, negative skin friction, soil arching
Procedia PDF Downloads 371918 Potential of Polyphenols from Tamarix Gallica towards Common Pathological Features of Diabetes and Alzheimer’s Diseases
Authors: Asma Ben Hmidene, Mizuho Hanaki, Kazuma Murakami, Kazuhiro Irie, Hiroko Isoda, Hideyuki Shigemori
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Type 2 diabetes mellitus (T2DM) and Alzheimer’s disease (AD) are characterized as a peripheral metabolic disorder and a degenerative disease of the central nervous system, respectively. It is now widely recognized that T2DM and AD share many pathophysiological features including glucose metabolism, increased oxidative stress and amyloid aggregation. Amyloid beta (Aβ) is the components of the amyloid deposits in the AD brain and while the component of the amyloidogenic peptide deposit in the pancreatic islets of Langerhans is identified as human islet amyloid polypeptide (hIAPP). These two proteins are originated from the amyloid precursor protein and have a high sequence similarity. Although the amino acid sequences of amyloidogenic proteins are diverse, they all adopt a similar structure in aggregates called cross-beta-spine. Add at that, extensive studies in the past years have found that like Aβ1-42, IAPP forms early intermediate assemblies as spherical oligomers, implicating that these oligomers possess a common folding pattern or conformation. These similarities can be used in the search for effective pharmacotherapy for DM, since potent therapeutic agents such as antioxidants with a catechol moiety, proved to inhibit Aβ aggregation, may play a key role in the inhibit the aggregation of hIAPP treatment of patients with DM. Tamarix gallica is one of the halophyte species having a powerful antioxidant system. Although it was traditionally used for the treatment of various liver metabolic disorders, there is no report about the use of this plant for the treatment or prevention of T2DM and AD. Therefore, the aim of this work is to investigate their protective effect towards T2DM and AD by isolation and identification of α-glucosidase inhibitors, with antioxidant potential, that play an important role in the glucose metabolism in diabetic patient, as well as, the polymerization of hIAPP and Aβ aggregation inhibitors. Structure-activity relationship study was conducted for both assays. And as for α-glucosidase inhibitors, their mechanism of action and their synergistic potential when applied with a very low concentration of acarbose were also suggesting that they can be used not only as α-glucosidase inhibitors but also be combined with established α-glucosidase inhibitors to reduce their adverse effect. The antioxidant potential of the purified substances was evaluated by DPPH and SOD assays. Th-T assay using 42-mer amyloid β-protein (Aβ42) for AD and hIAPP which is a 37-residue peptide secreted by the pancreatic β –cells for T2DM and Transmission electronic microscopy (TEM) were conducted to evaluate the amyloid aggragation of the actives substances. For α-glucosidase, p-NPG and glucose oxidase assays were performed for determining the inhibition potential and structure-activity relationship study. The Enzyme kinetic protocol was used to study the mechanism of action. From this research, it was concluded that polyphenols playing a role in the glucose metabolism and oxidative stress can also inhibit the amyloid aggregation, and that substances with a catechol and glucuronide moieties inhibiting amyloid-β aggregation, might be used to inhibit the aggregation of hIAPP.Keywords: α-glucosidase inhibitors, amyloid aggregation inhibition, mechanism of action, polyphenols, structure activity relationship, synergistic potential, tamarix gallica
Procedia PDF Downloads 270917 Capture of Co₂ From Natural Gas Using Modified Imidazolium Ionic Liquids
Authors: Alaa A. Ghanem, S. E. M. Desouky
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Natural gas (NG) is considered one of the most essential global energy sources. NG fields are often far away from the market, and a long-distance transporting pipeline usually is required. Production of NG with high content of CO₂ leads to severe problems such as equipment corrosion along with the production line until refinery.in addition to a high level of toxicity and decreasing in calorific value of the NG. So it is recommended to remove or decrease the CO₂ percent to meet transport specifications. This can be reached using different removal techniques such as physical and chemical absorption, pressure swing adsorption, membrane separation, or low-temperature separation. Many solvents and chemicals are being used to capture carbon dioxide on a large scale; among them, Ionic liquids have great potential due to their tunable properties; low vapour pressure, low melting point, and sensible thermal stability. In this research, three modifiedimidazolium ionic liquids will be synthesized and characterized using different tools of analysis such as FT-IR, 1H NMR. Thermal stability and surface activity will be studied. The synthesized compounds will be evaluated as selective solvents for CO₂ removal from natural gas using PVT cell.Keywords: natural gas, CO₂ capture, imidazolium ionic liquid, PVT cell
Procedia PDF Downloads 164916 Study of the Chemical Composition of Rye, Millet and Sorghum from Algeria
Authors: Soualem Mami Zoubida, Brixi Nassima, Beghdad Choukri, Belarbi Meriem
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Cereals are the most important source of dietary fiber in the Nordic diet. The fiber in cereals is located mainly in the outer layers of the kernel; particularly in the bran. Improved diet can help unlock the door to good health. Whole grains are an important source of nutrients that are in short supply in our diet, including digestible carbohydrates, dietary fiber, trace minerals, and other compounds of interest in disease prevention, including phytoestrogens and antioxidants (1). The objective of this study is to know the composition of whole grain cereals (rye, millet, white, and red sorghum) which a majority pushes in the south of Algeria. This shows that the millet has a high rate of the sugar estimated at 67.6%. The high proportion of proteins has been found in the two varieties of sorghum and rye. The millet presents the great percentage in lipids compared with the others cereals. And at the last, a red sorghum has the highest rate of fiber(2). These nutrients, as well as other components of whole grain cereals, have, in terms of health, an increased effect if they are consumed together.Keywords: chemical composition, miller, Secale cereal, Sorghum bicolor
Procedia PDF Downloads 403915 Ultrafiltration Process Intensification for Municipal Wastewater Reuse: Water Quality, Optimization of Operating Conditions and Fouling Management
Authors: J. Yang, M. Monnot, T. Eljaddi, L. Simonian, L. Ercolei, P. Moulin
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The application of membrane technology to wastewater treatment has expanded rapidly under increasing stringent legislation and environmental protection requirements. At the same time, the water resource is becoming precious, and water reuse has gained popularity. Particularly, ultrafiltration (UF) is a very promising technology for water reuse as it can retain organic matters, suspended solids, colloids, and microorganisms. Nevertheless, few studies dealing with operating optimization of UF as a tertiary treatment for water reuse on a semi-industrial scale appear in the literature. Therefore, this study aims to explore the permeate water quality and to optimize operating parameters (maximizing productivity and minimizing irreversible fouling) through the operation of a UF pilot plant under real conditions. The fully automatic semi-industrial UF pilot plant with periodic classic backwashes (CB) and air backwashes (AB) was set up to filtrate the secondary effluent of an urban wastewater treatment plant (WWTP) in France. In this plant, the secondary treatment consists of a conventional activated sludge process followed by a sedimentation tank. The UF process was thus defined as a tertiary treatment and was operated under constant flux. It is important to note that a combination of CB and chlorinated AB was used for better fouling management. The 200 kDa hollow fiber membrane was used in the UF module, with an initial permeability (for WWTP outlet water) of 600 L·m-2·h⁻¹·bar⁻¹ and a total filtration surface of 9 m². Fifteen filtration conditions with different fluxes, filtration times, and air backwash frequencies were operated for more than 40 hours of each to observe their hydraulic filtration performances. Through comparison, the best sustainable condition was flux at 60 L·h⁻¹·m⁻², filtration time at 60 min, and backwash frequency of 1 AB every 3 CBs. The optimized condition stands out from the others with > 92% water recovery rates, better irreversible fouling control, stable permeability variation, efficient backwash reversibility (80% for CB and 150% for AB), and no chemical washing occurrence in 40h’s filtration. For all tested conditions, the permeate water quality met the water reuse guidelines of the World Health Organization (WHO), French standards, and the regulation of the European Parliament adopted in May 2020, setting minimum requirements for water reuse in agriculture. In permeate: the total suspended solids, biochemical oxygen demand, and turbidity were decreased to < 2 mg·L-1, ≤ 10 mg·L⁻¹, < 0.5 NTU respectively; the Escherichia coli and Enterococci were > 5 log removal reduction, the other required microorganisms’ analysis were below the detection limits. Additionally, because of the COVID-19 pandemic, coronavirus SARS-CoV-2 was measured in raw wastewater of WWTP, UF feed, and UF permeate in November 2020. As a result, the raw wastewater was tested positive above the detection limit but below the quantification limit. Interestingly, the UF feed and UF permeate were tested negative to SARS-CoV-2 by these PCR assays. In summary, this work confirms the great interest in UF as intensified tertiary treatment for water reuse and gives operational indications for future industrial-scale production of reclaimed water.Keywords: semi-industrial UF pilot plant, water reuse, fouling management, coronavirus
Procedia PDF Downloads 109914 Computer Aided Screening of Secreted Frizzled-Related Protein 4 (SFRP4): A Potential Control for Diabetes Mellitus
Authors: Shazia Anwer Bukhari, Waseem Akhtar Shamshari, Mahmood-Ur-Rahman, Muhammad Zia-Ul-Haq, Hawa Z. E. Jaafar
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Diabetes mellitus is a life threatening disease and scientists are doing their best to find a cost effective and permanent treatment of this malady. The recent trend is to control the disease by target base inhibiting of enzymes or proteins. Secreted frizzled-related protein 4 (SFRP4) is found to cause five times more risk of diabetes when expressed above average levels. This study was therefore designed to analyze the SFRP4 and to find its potential inhibitors. SFRP4 was analyzed by bio-informatics tools of sequence tool and structure tool. A total of three potential inhibitors of SFRP4 were found, namely cyclothiazide, clopamide and perindopril. These inhibitors showed significant interactions with SFRP4 as compared to other inhibitors as well as control (acetohexamide). The findings suggest the possible treatment of diabetes mellitus type 2 by inhibiting the SFRP4 using the inhibitors cyclothiazide, clopamide and perindopril.Keywords: bioscreening, clopamide, cyclothiazide, diabetes mellitus, perindopril, SFRP4
Procedia PDF Downloads 440913 The Management of Behcet's Disease Patient's Mandibular Total Edentulism with Custom Made Implant Supported Bar Retainer: A Case Report
Authors: Faruk Emir, Simel Ayyıldız, Cem Şahin
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Behçet’s disease or Behçet’s syndrome is a chronic and multi-systemic inflammatory disease of unknown cause. This syndrome often presents with mucous membrane ulceration and ocular problems. As a systemic disease Behcet includes triple-symptom complex of recurrent oral aphthous ulcers, genital ulcers, and uveitis. Nearly all patients present with some form of painful oral mucocutaneous ulcerations in the form of aphthous ulcers. The aim of the treatment plan for Behçet’s Disease patients is to eliminate oral problems and increase the patient comfort.This clinical report represents the prosthodontic rehabilitation of Behcet’s disease patients mandibular total edentulism with the use of implant supported prosthesis that planned on custom abutments and bar retainers via CAD/CAM technology and patient satisfaction has been achieved in function and aesthetics.Keywords: Behçet’s disease, CAD/CAM, custom-made manufacturing, titanium milled bar retainer
Procedia PDF Downloads 320912 Android – Based Wireless Electronic Stethoscope
Authors: Aw Adi Arryansyah
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Using electronic stethoscope for detecting heartbeat sound, and breath sounds, are the effective way to investigate cardiovascular diseases. On the other side, technology is growing towards mobile. Almost everyone has a smartphone. Smartphone has many platforms. Creating mobile applications also became easier. We also can use HTML5 technology to creating mobile apps. Android is the most widely used type. This is the reason for us to make a wireless electronic stethoscope based on Android mobile. Android based Wireless Electronic Stethoscope designed by a simple system, uses sound sensors mounted membrane, then connected with Bluetooth module which will send the heart auscultation voice input data by Bluetooth signal to an android platform. On the software side, android will read the voice input then it will translate to beautiful visualization and release the voice output which can be regulated about how much of it is going to be released. We can change the heart beat sound into BPM data, and heart beat analysis, like normal beat, bradycardia or tachycardia.Keywords: wireless, HTML 5, auscultation, bradycardia, tachycardia
Procedia PDF Downloads 339911 Cell Surface Display of Xylanase on Escherichia coli by TibA Autotransporter
Authors: Yeng Min Yi, Rosli Md Illias, Salehhuddin Hamdan
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Industrial biocatalysis is mainly based on the use of cell free or intracellular enzyme systems. However, the expensive cost and relatively lower operational stability of free enzymes limit practical use in industries. Cell surface display system can be used as a cost-efficient alternative to overcome the laborious purification and substrate transport limitation. In this research, TibA autotransporter from E. coli was used to display Aspergillus fumigatus xylanase (xyn). The amplified xyn was fused in between N-terminal signal peptide and C-terminal β-barrel of TibA. The cloned was transformed and expressed in E. coli BL21 (DE3). Outer membrane localization of TibA-xyn fusion protein was confirmed by SDS PAGE and western blot with expected size of 62.5 kDa. Functional display of xyn was examined by activity assay. Cell surface displayed xyn exhibited the highest activity at 37 °c, 0.3 mM IPTG. As a summary, TibA displaying system has the potential for further industrial applications. Moreover, this is the first report of the display of xylanase using TibA on the surface of E. coli.Keywords: biocatalysis, cell surface display, Escherichia coli, TibA autotransporter
Procedia PDF Downloads 270910 Lifetime Assessment for Test Strips of POCT Device through Accelerated Degradation Test
Authors: Jinyoung Choi, Sunmook Lee
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In general, single parameter, i.e. temperature, as an accelerating parameter is used to assess the accelerated stability of Point-of-Care Testing (POCT) diagnostic devices. However, humidity also plays an important role in deteriorating the strip performance since major components of test strips are proteins such as enzymes. 4 different Temp./Humi. Conditions were used to assess the lifetime of strips. Degradation of test strips were studied through the accelerated stability test and the lifetime was assessed using commercial POCT products. The life distribution of strips, which were obtained by monitoring the failure time of test strip under each stress condition, revealed that the weibull distribution was the most proper distribution describing the life distribution of strips used in the present study. Equal shape parameters were calculated to be 0.9395 and 0.9132 for low and high concentrations, respectively. The lifetime prediction was made by adopting Peck Eq. Model for Stress-Life relationship, and the B10 life was calculated to be 70.09 and 46.65 hrs for low and high concentrations, respectively.Keywords: accelerated degradation, diagnostic device, lifetime assessment, POCT
Procedia PDF Downloads 407