Search results for: mitochondrial membrane
900 Experimental and Characterization Studies on Micro Direct Methanol Fuel Cell
Authors: S. Muthuraja Soundrapandian, C.K. Subramaniam
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A micro Direct Methanol Fuel Cell (DMFC) of 1 cm2 active area with selective sensor materials to sense methanol for redox, has been developed. Among different Pt alloys, Pt-Sn/C was able to produce high current density and repeatability. Membrane Elecctrode Assembly (MEA) of anode catalyst Pt-Sn/C was prepared with nafion as active membrane and Pt black as cathode catalyst. The sensor’s maximum ability to detect the trace levels of methanol in ppm has been analyzed. A compact sensor set up has also been made and the characterization studies were carried out. The acceptable value of current density was derived by the cell and the results are able to fulfill the needs of DMFC technology for the practical applications.Keywords: DMFC, sensor, MEA, Pt-Sn
Procedia PDF Downloads 140899 Formation of Nanochannels by Heavy Ions in Graphene Oxide Reinforced Carboxymethylcellulose Membranes for Proton Exchange Membrane Fuel Cells Applications
Authors: B. Kurbanova, M. Karibayev, N. Almas, K. Ospanov, K. Aimaganbetov, T. Kuanyshbekov, K. Akatan, S. Kabdrakhmanova
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Proton exchange membranes (PEMs) operating at high temperatures above 100 °C with the excellent mechanical, chemical and thermochemical stability have been received much attention, because of their practical application of proton exchange membrane fuel cells (PEMFCs). Nowadays, a huge number of polymers and polymer-mixed various membranes have been investigated for this application, all of which offer both pros and cons. However, PEMFCs are still lack of ideal membranes with unique properties. In this work, carboxymethylcellulose (CMC) based membranes with dispersive graphene oxide (GO) sheets were fabricated and investigated for PEMFCs application. These membranes and pristine GO were studied by a combination of XRD, XPS, Raman, Brillouin, FTIR, thermo-mechanical analysis (TGA and Dynamic Mechanical Analysis) and SEM microscopy, while substantial studies on the proton transport properties were provided by Electrochemical Impedance Spectroscopy (EIS) measurements. It was revealed that the addition of CMC to the GO boosts proton conductivity of the whole membrane, while GO provides good mechanical and thermomechanical stability to the membrane. Further, the continuous and ordered nanochannels with well-tailored chemical structures were obtained by irradiation of heavy ions Kr⁺¹⁷ with an energy of 1.75 MeV/nucleon on the heavy ion accelerator. The formation of these nanochannels led to the significant increase of proton conductivity at 50% Relative Humidity. Also, FTIR and XPS measurement results show that ion irradiation eliminated the GO’s surface oxygen chemical bonds (C=O, C-O), and led to the formation of C = C, C – C bonds, whereas these changes connected with an increase in conductivity.Keywords: proton exchange membranes, graphene oxide, fuel cells, carboxymethylcellulose, ion irradiation
Procedia PDF Downloads 92898 Thin-Film Nanocomposite Membrane with Single-Walled Carbon Nanotubes Axial Positioning in Support Layer for Desalination of Water
Authors: Ahmed A. Alghamdi
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Single-walled carbon nanotubes (SWCNTs) are an outstanding material for applications in thermoelectric power generation, nanoelectronics, electrochemical energy storage, photovoltaics, and light emission. They are ultra-lightweight and possess electrical as well as thermal conductivity, flexibility, and mechanical strength. SWCNT is applicable in water treatment, brine desalination, removal of heavy metal ions associated with pollutants, and oil-water separation. Carbon nanotube (CNT) is believed to tackle the trade-off issue between permeability, selectivity, and fouling issues in membrane filtration applications. Studying these CNT structures, as well as their interconnection in nanotechnology, assists in finding the precise position to be placed for water desalination. Reverse osmosis (RO) has been used globally for desalination, resulting in purified water. Thin film composite (TFC) membranes were utilized in the RO process for desalination. The sheet thickness increases the salt rejection and decreases the water flux when CNT is utilized as a support layer to this membrane. Thus, through a temperature-induced phase separation technique (TIPS), axially aligned SWCNT (AASWCNT) is fabricated, and its use enhances the salt rejection and water flux at short reaction times with a modified procedure. An evaluation was conducted and analogized with prior works in the literature, which exhibited that the prepared TFC membrane showed a better outcome.Keywords: single-walled carbon nanotubes, thin film composite, axially aligned swcnt, temperature induced phase separation technique, reverse osmosis
Procedia PDF Downloads 51897 Membrane Permeability of Middle Molecules: A Computational Chemistry Approach
Authors: Sundaram Arulmozhiraja, Kanade Shimizu, Yuta Yamamoto, Satoshi Ichikawa, Maenaka Katsumi, Hiroaki Tokiwa
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Drug discovery is shifting from small molecule based drugs targeting local active site to middle molecules (MM) targeting large, flat, and groove-shaped binding sites, for example, protein-protein interface because at least half of all targets assumed to be involved in human disease have been classified as “difficult to drug” with traditional small molecules. Hence, MMs such as peptides, natural products, glycans, nucleic acids with various high potent bioactivities become important targets for drug discovery programs in the recent years as they could be used for ‘undruggable” intracellular targets. Cell membrane permeability is one of the key properties of pharmacodynamically active MM drug compounds and so evaluating this property for the potential MMs is crucial. Computational prediction for cell membrane permeability of molecules is very challenging; however, recent advancement in the molecular dynamics simulations help to solve this issue partially. It is expected that MMs with high membrane permeability will enable drug discovery research to expand its borders towards intracellular targets. Further to understand the chemistry behind the permeability of MMs, it is necessary to investigate their conformational changes during the permeation through membrane and for that their interactions with the membrane field should be studied reliably because these interactions involve various non-bonding interactions such as hydrogen bonding, -stacking, charge-transfer, polarization dispersion, and non-classical weak hydrogen bonding. Therefore, parameters-based classical mechanics calculations are hardly sufficient to investigate these interactions rather, quantum mechanical (QM) calculations are essential. Fragment molecular orbital (FMO) method could be used for such purpose as it performs ab initio QM calculations by dividing the system into fragments. The present work is aimed to study the cell permeability of middle molecules using molecular dynamics simulations and FMO-QM calculations. For this purpose, a natural compound syringolin and its analogues were considered in this study. Molecular simulations were performed using NAMD and Gromacs programs with CHARMM force field. FMO calculations were performed using the PAICS program at the correlated Resolution-of-Identity second-order Moller Plesset (RI-MP2) level with the cc-pVDZ basis set. The simulations clearly show that while syringolin could not permeate the membrane, its selected analogues go through the medium in nano second scale. These correlates well with the existing experimental evidences that these syringolin analogues are membrane-permeable compounds. Further analyses indicate that intramolecular -stacking interactions in the syringolin analogues influenced their permeability positively. These intramolecular interactions reduce the polarity of these analogues so that they could permeate the lipophilic cell membrane. Conclusively, the cell membrane permeability of various middle molecules with potent bioactivities is efficiently studied using molecular dynamics simulations. Insight of this behavior is thoroughly investigated using FMO-QM calculations. Results obtained in the present study indicate that non-bonding intramolecular interactions such as hydrogen-bonding and -stacking along with the conformational flexibility of MMs are essential for amicable membrane permeation. These results are interesting and are nice example for this theoretical calculation approach that could be used to study the permeability of other middle molecules. This work was supported by Japan Agency for Medical Research and Development (AMED) under Grant Number 18ae0101047.Keywords: fragment molecular orbital theory, membrane permeability, middle molecules, molecular dynamics simulation
Procedia PDF Downloads 189896 Identifying Apis millefera Strains in Akkar District (North Lebanon) Using Mitochondrial DNA: A Step in Preserving the Local Strain A. m. Syriaca
Authors: Zeina Nasr, Bashar Merheb
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The honey bee is a social insect that had driven the human interest much more than any other organism. Beekeeping practices dated the appearance of Man on earth and now it provides a hobby or a secondary work that contributes to the family revenue and requires a little time engagement and money investment. Honey production is not the only contribution of honey bees to the economy, since honey bees play an important role in the pollination. Bee keeping in Lebanon is an important part of the agricultural economy. However, a growing concern about bees is spreading globally, due to an accelerated decline of bees colonies. This raises the alert to preserve and protect local bee strains against uncontrolled introduction of foreign strains and invasive parasitic species. Mitochondrial DNA (mtDNA) markers are commonly used in studying genetic variation in the Apis mellifera species. The DraI-COI-COII test is based on the analysis of the intergenic region between the two genes COI and COII. The different honey bee strains differ in the presence or absence of the p sequence and the number of Q sequences present. A. m. syriaca belonging to the lineage Z, is the native honey bee subspecies in Lebanon, Syria, Jordan, and Palestine. A. m. syriaca is known for its high defensiveness, even though it has many important advantages. However, commercial breeder strains, such as the Italian (A. m. ligustica), and Carniolan (A. m. carnica) strains, have been introduced by beekeepers and regularly used for honey production. This raises worries about the disappearance of the local subspecies. It is obvious that identifying A. m. syriaca colonies and protecting them against uncontrolled mating with other bee strains is a crucial step to protect and improve the original local strain. This study aims to reveal the existing sub-species of honey bee in Akkar – Lebanon and to assess the influence of introgression on the hybridization of the local strain. This will help to identify areas of pure A.m. syriaca population over this district to be considered in choosing syriaca reserves. We collected samples of bees from different regions of Akkar district in order to perform mtDNA analysis. We determined the restriction fragments length of the intergenic region COI-COII, using the restriction enzyme DraI. The results showed both the C and the Z lineages. Four restriction patterns were identified among the restriction maps of the studied samples. The most abundant mitochondrial lineage is the Z lineage constituting about 60% of the identified samples. Al-Dreib region reported the lowest introgression with foreign mtDNA of 21% making it the most suitable area for a genetic reserve of syriaca in Akkar based on its lowest introgression and suitable environment in addition to the attitude of local beekeepers to conserve the local strain. Finally, this study is the first step in constructing conservation programs for the preservation of the local strain and should be generalized to the whole Lebanese population, consistent with the effort done in neighboring countries.Keywords: Akkar Lebanon, Apis millefera syriaca, DraI-COI-COII test, mitochondrial DNA
Procedia PDF Downloads 276895 Extracellular Polymeric Substances (EPS) Attribute to Biofouling of Anaerobic Membrane Bioreactor: Adhesion and Viscoelastic Properties
Authors: Kbrom Mearg Haile
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Introduction: Membrane fouling is the bottleneck for the anaerobic membrane bioreactor (AnMBR) robust continuous operation, primarily caused by the mixed liquor suspended solids (MLSS) characteristics formed by aggregated flocs and a scaffold of microbial self-produced extracellular polymeric substances (EPS), which dictates the flocs integrity. Accordingly, the adhesion of EPS to the membrane surface versus their role in forming firm, elastic, and mechanically stable flocs under the reactor’s hydraulic shear is critical for minimizing interactions between EPS and colloids originating from the MLSS flocs with the membrane. This study aims to gain insight and investigate the effect of MLSS flocs properties, EPS adhesion and viscoelasticity, viscoelastic properties of the sludge, and membrane fouling propensity. Experimental: As a working hypothesis, to alter the aforementioned flocs’ and EPS’s properties, the addition of either coagulant or surfactant was carried out during the AnMBR operation. In the AnMBR, two flat-sheet 300 kDa pore size polyether sulfone (PES) membranes with a total filtration area of 352 cm2 were immersed in the AnMBR system treating municipal wastewater of Midreshet Ben-Gurion village at the Negev highlands, Israel. The system temperature, pH, biogas recirculation, and hydraulic retention time were regulated. TMP fluctuations during a 30-day experiment were recorded under three operating conditions: Baseline (without the addition of coagulating or dispersing agent), coagulant addition (FeCl3), and surfactant addition (sodium dodecyl sulfate). At the end of each experiment, EPS were extracted from the MLSS and from the fouled membrane, characterized for their protein, polysaccharides, and DOC contents, and correlated with the fouling tendency of the submerged UF membrane. The EPS adherence and viscoelastic properties were revealed using QCM-D via the PES-coated gold sensor used as a membrane-mimicking surface providing a detailed real-time EPS adhesion. The associated shifts in the resonance frequency and dissipation at different overtones were further modeled using the Voigt-based viscoelastic model (using Dfind software, Q-Sense Biolin Scientific) in which the thickness, shear modulus, and shear viscosity values of the adsorbed EPS layers on the PES coated sensor were calculated. Results and discussion: The observations obtained from the QCM-D analysis indicate a greater decrease in the frequency shift for the elevated membrane fouling scenarios, likely due to an observed decrease in the calculated shear viscosity and shear modulus of the EPS adsorbed layer, coupled with an increase in EPS layer hydrated thickness and fluidity (ΔD/Δf slopes). Further analysis is being conducted for the three major operating conditions-analyzing their effects on sludge rheology, dewaterability (capillary suction time-CST) and settle ability (SVI). The biofouling layer is further characterized microscopically using a confocal laser scanning microscope (CLSM) and scanning electron microscope (SEM), for analyzing the consistency of the development of the biofouling layer with sludge characteristics, i.e., thicker biofouling layer on the membrane surface when operated with surfactant addition, due to flocs with reduced integrity and availability of EPS/colloids to the membrane. Conversely, a thinner layer when operated with coagulant compared to the baseline experiment, due to elevation in flocs integrity.Keywords: viscoelasticity, biofouling, viscoelastic, AnMBR, EPS, elocintegrity
Procedia PDF Downloads 22894 Quality of Chilled Indigenous Ram Semen Using Multi-Species Skim Milk Based Extenders
Authors: Asaduzzaman Rimon, Pankaj Kumar Jha, Abdullah Al Mansur, Mohammad Mofizul Islam, Nasrin Sultana Juyena, Farida Yeasmin Bari
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This study was conducted to determine the effects of multi-species skim milk based extenders on sperm quality at 5ºC with the advancement of preservation time. Altogether forty ejaculates, 8 ejaculates for each of the 5 home-made semen extenders: cow skim milk (CSM), goat skim milk (GSM), sheep skim milk (SSM), buffalo skim milk (BSM) and commercial dried skim milk (CDSM) were examined for motility, plasma membrane integrity and normal morphology % of sperm at 0, 24, 48, 72, 96 and 120 hours, respectively. Sperm motility was significantly decreased (P < 0.05) with the increase of preservation time. There were no significant difference in motility % among CSM (84.0±1.4, 82.3±2.1), GSM (84.5±1.0, 82.5±0.6) and CDSM (85.0±80.3±1.3) extenders at 0 and 24 hours, respectively. However, the motility in GSM extender was significantly higher than BSM, SSM and CDSM extender at 48, 72, 96 and 120 hours. The plasma membrane integrity % at 0 hour had no significant difference among the extenders. But, the plasma membrane integrity % in GSM (84.3±0.9, 81.8±1.3, 78.0±2.2, 74.8±0.5, 72.0±1.4) and CSM (82.8±0.5, 80.8±1.0, 78.0±1.4, 73.5±1.7, 70.3±0.5) extenders were significantly higher than BSM (81.0±1.4, 76.3±2.5, 72.5±1.7, 63.8±2.5, 54.0±4.6), SSM (78.5±1.5, 75.0±1.6, 71.5±2.4, 64.3±1.7, 56.5±2.4) and CDSM extenders (78.3±2.4, 75.8±3.9, 72.5±3.3, 64.8±1.0, 60.5±3.3) at 24, 48, 72, 96 and 120 hours, respectively. The sperm morphology % had no significant difference at 0 hour among the extenders but were significantly higher in GSM (83.0±0.8, 81.3±1.5, 79.3±1.3, 73.0±2.2, 70.3±1.3) and CSM (81.5±1.7, 79.3±1.5, 75.8±1.5, 70.3±1.3, 66.3±1.5) than BSM (79.0±1.2, 75.0±1.4, 69.5±1.7, 64.5±3.1, 56.8±2.2), SSM (79.8±1.3, 76.8±2.1, 71.3±3.0, 66.0±2.7, 60.3±4.5) and CDSM (80.0±1.6, 77.0±2.2, 72.0±2.5, 66.3±2.5, 62.0±4.0) extenders at 24, 48, 72, 96 and 120 hours, respectively. The motility, plasma membrane integrity and normal morphology % of sperm had shown no significant difference between GSM and CSM but were found to be higher in GSM extenders. In the end, we concluded from the above study that the goat milk based extenders (GSM) had optimum sperm preserving quality. However, further studies are required to validate followed by fertility rate.Keywords: chilled semen, indigenous ram, multi-species skim milk based extenders, preservation
Procedia PDF Downloads 421893 In-situ Oxygen Enrichment for UCG
Authors: Adesola O. Orimoloye, Edward Gobina
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Membrane separation technology is still considered as an emerging technology in the mining sector and does not yet have the widespread acceptance that it has in other industrial sectors. Underground Coal Gasification (UCG), wherein coal is converted to gas in-situ, is a safer alternative to mining method that retains all pollutants underground making the process environmentally friendly. In-situ combustion of coal for power generation allows access to more of the physical global coal resource than would be included in current economically recoverable reserve estimates. Where mining is no longer taking place, for economic or geological reasons, controlled gasification permits exploitation of the deposit (again a reaction of coal to form a synthesis gas) of coal seams in situ. The oxygen supply stage is one of the most expensive parts of any gasification project but the use of membranes is a potentially attractive approach for producing oxygen-enriched air. In this study, a variety of cost-effective membrane materials that gives an optimal amount of oxygen concentrations in the range of interest was designed and tested at diverse operating conditions. Oxygen-enriched atmosphere improves the combustion temperature but a decline is observed if oxygen concentration exceeds optimum. Experimental result also reveals the preparatory method, apparatus and performance of the fabricated membrane.Keywords: membranes, oxygen-enrichment, gasification, coal
Procedia PDF Downloads 322892 Bimetallic MOFs Based Membrane for the Removal of Heavy Metal Ions from the Industrial Wastewater
Authors: Muhammad Umar Mushtaq, Muhammad Bilal Khan Niazi, Nouman Ahmad, Dooa Arif
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Apart from organic dyes, heavy metals such as Pb, Ni, Cr, and Cu are present in textile effluent and pose a threat to humans and the environment. Many studies on removing heavy metallic ions from textile wastewater have been conducted in recent decades using metal-organic frameworks (MOFs). In this study new polyether sulfone ultrafiltration membrane, modified with Cu/Co and Cu/Zn-based bimetal-organic frameworks (MOFs), was produced. Phase inversion was used to produce the membrane, and atomic force microscopy (AFM), scanning electron microscopy (SEM) were used to characterize it. The bimetallic MOFs-based membrane structure is complex and can be comprehended using characterization techniques. The bimetallic MOF-based filtration membranes are designed to selectively adsorb specific contaminants while allowing the passage of water molecules, improving the ultrafiltration efficiency. MOFs' adsorption capacity and selectivity are enhanced by functionalizing them with particular chemical groups or incorporating them into composite membranes with other materials, such as polymers. The morphology and performance of the bimetallic MOF-based membrane were investigated regarding pure water flux and metal ion rejection. The advantages of developed bimetallic MOFs based membranes for wastewater treatment include enhanced adsorption capacity because of the presence of two metals in their structure, which provides additional binding sites for contaminants, leading to a higher adsorption capacity and more efficient removal of pollutants from wastewater. Based on the experimental findings, bimetallic MOF-based membranes are more capable of rejecting metal ions from industrial wastewater than conventional membranes that have already been developed. Furthermore, the difficulties associated with operational parameters, including pressure gradients and velocity profiles, are simulated using Ansys Fluent software. The simulation results obtained for the operating parameters are in complete agreement with the experimental results.Keywords: bimetallic MOFs, heavy metal ions, industrial wastewater treatment, ultrafiltration.
Procedia PDF Downloads 90891 Passive Heat Exchanger for Proton Exchange Membrane Fuel Cell Cooling
Authors: Ivan Tolj
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Water produced during electrochemical reaction in Proton Exchange Membrane (PEM) fuel cell can be used for internal humidification of reactant gases; hydrogen and air. On such a way it is possible to eliminate expensive external humidifiers and simplify fuel cell balance-of-plant (BoP). When fuel cell operates at constant temperature (usually between 60 °C and 80 °C) relatively cold and dry ambient air heats up quickly upon entering channels which cause further drop in relative humidity (below 20%). Low relative humidity of reactant gases dries up polymer membrane and decrease its proton conductivity which results in fuel cell performance drop. It is possible to maintain such temperature profile throughout fuel cell cathode channel which will result in close to 100 % RH. In order to achieve this, passive heat exchanger was designed using commercial CFD software (ANSYS Fluent). Such passive heat exchanger (with variable surface area) is suitable for small scale PEM fuel cells. In this study, passive heat exchanger for single PEM fuel cell segment (with 20 x 1 cm active area) was developed. Results show close to 100 % RH of air throughout cathode channel with increased fuel cell performance (mainly improved polarization curve) and improved durability.Keywords: PEM fuel cell, passive heat exchange, relative humidity, thermal management
Procedia PDF Downloads 277890 Collection, Cryopreservation, and Fertilizing Potential of Bovine Spermatozoa Collected from the Epididymis Evaluated by Conventional Techniques and by Flow Cytometry
Authors: M. H. Moreira da Silva, L. Valadao, F. Moreira da Silva
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In the present study, the fertilizing capacity of bovine spermatozoa was evaluated before and after its cryopreservation. For this, the testicles of 100 bulls slaughtered on Terceira Island were dissected, the epididymal tails were separated, and semen was recovered by the flotation method and then evaluated by phase contrast microscopy and by flow cytometry. For phase contrast microscopy, a drop of semen was used to evaluate the percentage of motile spermatozoa (from 0 to 100%) and motility (from 0 to 5). After determining the concentration and the abnormal forms, semen was diluted to a final concentration of 50 x 106 spz/ml and evaluated by flow cytometer for membrane and acrosome integrity using the conjugation of fluorescent probes propidium iodide (PI) and Arachis hypogea agglutinin (FITC-PNA). Freezing was carried out in a programmable semen freezer, using 0.25 ml straws, in a total of 20 x 106 viable sperm per straw with glycerol as a cryoprotectant in a final concentration of 0.58 M. It was observed that, on average, a total of 7.25 ml of semen was collected from each bull. The viability and vitality rates were respectively 83.22 ± 7.52% and 3.8 ± 0.4 before freezing, decreasing to 58.81 ± 11.99% and 3.6 ± 0.6, respectively, after thawing. Regarding cytoplasmic droplets, it was observed that a high percentage of spermatozoa had medial cytoplasmic droplets (38.47%), with only 3.32% and 0.15% presenting proximal and distal cytoplasmic drops, respectively. By flow cytometry, it was observed that before freezing, the percentage of sperm with the damaged plasma membrane and intact acrosome was 3.61 ± 0.99%, increasing slightly to 4.21 ± 1.86% after cryopreservation (p<0.05). Regarding spermatozoa with damaged plasma membrane and acrosome, the percentage before freezing was 3.37±1.87%, increasing to 4.34 ±1.16% after thawing, and no significant differences were observed between these two values. For the percentage of sperm with the intact plasma membrane and damaged acrosome, this value was 2.04 ± 2.34% before freezing, decreasing to 0.89 ± 0.48% after thawing (p<0.05). The percentage of sperm with the intact plasma membrane and acrosome before freezing was 90.99±2.75%, with a slight decrease to 90.57±3.15% after thawing (p<0.05). From this study, it can be clearly concluded that, after the slaughtering of bulls, the spermatozoa can be recovered from the epididymis and cryopreserved, maintaining an excellent rate of sperm viability and quality after thawing.Keywords: bovine semen, epididymis, cryopreservation, fertility assessment
Procedia PDF Downloads 90889 Artificial Neural Network Reconstruction of Proton Exchange Membrane Fuel Cell Output Profile under Transient Operation
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Unbalanced power output from individual cells of Proton Exchange Membrane Fuel Cell (PEMFC) has direct effects on PEMFC stack performance, in particular under transient operation. In the paper, a multi-layer ANN (Artificial Neural Network) model Radial Basis Functions (RBF) has been developed for predicting cells' output profiles by applying gas supply parameters, cooling conditions, temperature measurement of individual cells, etc. The feed-forward ANN model was validated with experimental data. Influence of relevant parameters of RBF on the network accuracy was investigated. After adequate model training, the modelling results show good correspondence between actual measurements and reconstructed output profiles. Finally, after the model was used to optimize the stack output performance under steady-state and transient operating conditions, it suggested that the developed ANN control model can help PEMFC stack to have obvious improvement on power output under fast acceleration process.Keywords: proton exchange membrane fuel cell, PEMFC, artificial neural network, ANN, cell output profile, transient
Procedia PDF Downloads 169888 Self-Assembled Laser-Activated Plasmonic Substrates for High-Throughput, High-Efficiency Intracellular Delivery
Authors: Marinna Madrid, Nabiha Saklayen, Marinus Huber, Nicolas Vogel, Christos Boutopoulos, Michel Meunier, Eric Mazur
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Delivering material into cells is important for a diverse range of biological applications, including gene therapy, cellular engineering and imaging. We present a plasmonic substrate for delivering membrane-impermeable material into cells at high throughput and high efficiency while maintaining cell viability. The substrate fabrication is based on an affordable and fast colloidal self-assembly process. When illuminated with a femtosecond laser, the light interacts with the electrons at the surface of the metal substrate, creating localized surface plasmons that form bubbles via energy dissipation in the surrounding medium. These bubbles come into close contact with the cell membrane to form transient pores and enable entry of membrane-impermeable material via diffusion. We use fluorescence microscopy and flow cytometry to verify delivery of membrane-impermeable material into HeLa CCL-2 cells. We show delivery efficiency and cell viability data for a range of membrane-impermeable cargo, including dyes and biologically relevant material such as siRNA. We estimate the effective pore size by determining delivery efficiency for hard fluorescent spheres with diameters ranging from 20 nm to 2 um. To provide insight to the cell poration mechanism, we relate the poration data to pump-probe measurements of micro- and nano-bubble formation on the plasmonic substrate. Finally, we investigate substrate stability and reusability by using scanning electron microscopy (SEM) to inspect for damage on the substrate after laser treatment. SEM images show no visible damage. Our findings indicate that self-assembled plasmonic substrates are an affordable tool for high-throughput, high-efficiency delivery of material into mammalian cells.Keywords: femtosecond laser, intracellular delivery, plasmonic, self-assembly
Procedia PDF Downloads 530887 Wrinkling Prediction of Membrane Composite of Varying Orientation under In-Plane Shear
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In this article, the wrinkling failure of orthotropic composite membranes due to in-plane shear deformation is investigated using nonlinear finite element analyses. A nonlinear post-buckling analysis is performed to show the evolution of shear-induced wrinkles. The method of investigation is based on the post-buckling finite element analysis adopted from commercial FEM code; ANSYS. The resulting wrinkling patterns, their amplitude and their wavelengths under the prescribed loads and boundary conditions were confirmed by experimental results. Our study reveals that wrinkles develop when both the magnitudes and coverage of the minimum principal stresses in the laminated composite laminates are sufficiently large to trigger wrinkling.Keywords: composite, FEM, membrane, wrinkling
Procedia PDF Downloads 275886 Characterization of Carbon Dioxide-Rich Flue Gas Sources for Conversion to Chemicals and Fuels
Authors: Adesola Orimoloye, Edward Gobina
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Flue gas is the most prevalent source of carbon dioxide off-gas from numerous processes globally. Among the lion's share of this flue gas is the ever - present electric power plant, primarily fuelled by coal, and then secondly, natural gas. The carbon dioxide found in coal fired power plant off gas is among the dirtiest forms of carbon dioxide, even with many of the improvements in the plants; still this will yield sulphur and nitrogen compounds; among other rather nasty compounds and elements; all let to the atmosphere. This presentation will focus on the characterization of carbon dioxide-rich flue gas sources with a view of eventual conversion to chemicals and fuels using novel membrane reactors.Keywords: Flue gas, carbon dioxide, membrane, catalyst, syngas
Procedia PDF Downloads 674885 An Investigation into the Potential of Industrial Low Grade Heat in Membrane Distillation for Freshwater Production
Authors: Yehia Manawi, Ahmad Kayvanifard
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Membrane distillation is an emerging technology which has been used to produce freshwater and purify different types of aqueous mixtures. Qatar is an arid country where almost 100% of its freshwater demand is supplied through the energy-intensive thermal desalination process. The country’s need for water has reached an all-time high which stipulates finding an alternative way to augment freshwater without adding any drastic affect to the environment. The objective of this paper was to investigate the potential of using the industrial low grade waste heat to produce freshwater using membrane distillation. The main part of this work was conducting a heat audit on selected Qatari chemical industries to estimate the amounts of freshwater produced if such industrial waste heat were to be recovered. By the end of this work, the main objective was met and the heat audit conducted on the Qatari chemical industries enabled us to estimate both the amounts of waste heat which can be potentially recovered in addition to the amounts of freshwater which can be produced if such waste heat were to be recovered. By the end, the heat audit showed that around 605 Mega Watts of waste heat can be recovered from the studied Qatari chemical industries which resulted in a total daily production of 5078.7 cubic meter of freshwater. This water can be used in a wide variety of applications such as human consumption or industry. The amount of produced freshwater may look small when compared to that produced through thermal desalination plants; however, one must bear in mind that this water comes from waste and can be used to supply water for small cities or remote areas which are not connected to the water grid. The idea of producing freshwater from the two widely-available wastes (thermal rejected brine and waste heat) seems promising as less environmental and economic impacts will be associated with freshwater production which may in the near future augment the conventional way of producing freshwater currently being thermal desalination. This work has shown that low grade waste heat in the chemical industries in Qatar and perhaps the rest of the world can contribute to additional production of freshwater using membrane distillation without significantly adding to the environmental impact.Keywords: membrane distillation, desalination, heat recovery, environment
Procedia PDF Downloads 319884 Genetic Diversity and Variation of Nigerian Pigeon (Columba livia domestica) Populations Based on the Mitochondrial Coi Gene
Authors: Foluke E. Sola-Ojo, Ibraheem A. Abubakar, Semiu F. Bello, Isiaka H. Fatima, Sule Bisola, Adesina M. Olusegun, Adeniyi C. Adeola
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The domesticated pigeon, Columba livia domestica, has many valuable characteristics, including high nutritional value and fast growth rate. There is a lack of information on its genetic diversity in Nigeria; thus, the genetic variability in mitochondrial cytochrome oxidase subunit I (COI) sequences of 150 domestic pigeons from four different locations was examined. Three haplotypes (HT) were identified in Nigerian populations; the most common haplotype, HT1, was shared with wild and domestic pigeons from Europe, America, and Asia, while HT2 and HT3 were unique to Nigeria. The overall haplotype diversity was 0.052± 0.025, and nucleotide diversity was 0.026± 0.068 across the four investigated populations. The phylogenetic tree showed significant clustering and genetic relationship of Nigerian domestic pigeons with other global pigeons. The median-joining network showed a star-like pattern suggesting population expansion. AMOVA results indicated that genetic variations in Nigerian pigeons mainly occurred within populations (99.93%), while the Neutrality tests results suggested that the Nigerian domestic pigeons’ population experienced recent expansion. This study showed a low genetic diversity and population differentiation among Nigerian domestic pigeons consistent with a relatively conservative COI sequence with few polymorphic sites. Furthermore, the COI gene could serve as a candidate molecular marker to investigate the genetic diversity and origin of pigeon species. The current data is insufficient for further conclusions; therefore, more research evidence from multiple molecular markers is required.Keywords: Nigeria pigeon, COI, genetic diversity, genetic variation, conservation
Procedia PDF Downloads 195883 Region-Specific Secretory Protein, α2M, in Male Reproductive Tract of the Blue Crab And Its Dynamics during Sperm transit towards Female Spermatheca
Authors: Thanyaporn Senarai, Rapeepun Vanichviriyakit, Shinji Miyata, Chihiro Sato, Prapee Sretarugsa, Wattana Weerachatyanukul, Ken Kitajima
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In this study, we characterized a region-specific 250 kDa protein that was secreted of MSD fluid, which is believed to play dual functions in forming a spermatophoric wall for sperm physical protection, and in sperm membrane modification as part of sperm maturation process. The partial amino acid sequence and N-terminal sequencing revealed that the MSD-specific 250 kDa protein showed a high similarity with a plasma-rich protein, α-2 macroglobulin (α2M), so termed pp-α2M. This protein was a large glycoprotein contained predominantly mannose and GlcNAc. The expression of pp-α2M mRNA was detected in spermatic duct (SD), androgenic gland (AG) and hematopoietic tissue, while the protein expression was rather specific to the apical cytoplasm of MSD epithelium. The secretory pp-α2M in MSD fluid was acquired onto the MSD sperm membrane and was also found within the matrix of the acrosome. Distally, pp-α2M was removed from spermathecal sperm membrane, while its level kept constant in the sperm AC. Together the results indicate that pp-α2M is a 250 kDa region-specific secretory protein which plays roles in sperm physical protection and also acts as maturation factor in the P. pelagicus sperm.Keywords: alpha-2 macroglobulin, blue swimming crab, sperm maturation, spermatic duct
Procedia PDF Downloads 329882 Preparation and Characterizations of Natural Material Based Ceramic Membranes
Authors: In-Hyuck Song, Jang-Hoon Ha
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Recently, porous ceramic membranes have attracted great interest due to their outstanding thermal and chemical stability. In this paper, we report the results of our efforts to determine whether we could prepare a diatomite-kaolin composite coating to be deposited over a sintered diatomite support layer that could reduce the largest pore size of the sintered diatomite membrane while retaining an acceptable level of permeability. We determined under what conditions such a composite coating over a support layer could be prepared without the generation of micro-cracks during drying and sintering. The pore characteristics of the sintered diatomite membranes were studied by scanning electron microscopy and capillary flow porosimetry.Keywords: ceramic membrane, diatomite, water treatment, sintering
Procedia PDF Downloads 515881 Rapid Mitochondrial Reactive Oxygen Species Production Precedes NF-κB Activation and Pro-inflammatory Responses in Macrophages
Authors: Parinaz Tavakoli Zaniani, Dimitrios Balomenos
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Mitochondrial reactive oxygen species (mROS) play a crucial role in macrophage pro-inflammatory activation, although a detailed understanding of the mechanism and kinetics by which mROS drive signaling molecules is still lacking. In general, it is thought that NF-κB activation drives mROS and general ROS production. Here, We performed a detailed kinetic analysis of mROS production during macrophage activation. We found early mROS generation after LPS (lipopolysaccharide) stimulation. Remarkably as early as 5 minutes, mROS signaling promoted initial NF-κB, MAPK activation and pro-inflammatory cytokine production, as established through inhibition or quenching of mROS. On the contrary, NF-κB inhibition had no effect on mROS production. Our findings point to a mechanism by which mROS increase TRAF-6 ubiquitination and, thus NF-κB activity. mROS inhibition reduced LPS-induced lethality in an in vivo septic shock model by controlling pro-inflammatory cytokine production. Overall, our research provides novel insights into the role of mROS as a primary messenger in the pathway of macrophage and as a regulator of inflammatory responses. We found that early mROS production promotes initial NF-κB, and MAPK activation by regulating TRAF-6 ubiquitination and that mROS inhibition can reduce LPS-induced inflammatory cytokines and lethality in a septic shock model. These findings might lead to novel immunotherapeutic strategies targeting early mROS production and control of extreme inflammation in the context of sepsis and other inflammatory diseases.Keywords: mitochondria, reactive oxygen species, nuclear factor κB, lipopolysaccharide, macrophages
Procedia PDF Downloads 75880 In-situ Oxygen Enrichment for Underground Coal Gasification
Authors: Adesola O. Orimoloye, Edward Gobina
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Membrane separation technology is still considered as an emerging technology in the mining sector and does not yet have the widespread acceptance that it has in other industrial sectors. Underground Coal Gasification (UCG), wherein coal is converted to gas in-situ, is a safer alternative to mining method that retains all pollutants underground making the process environmentally friendly. In-situ combustion of coal for power generation allows access to more of the physical global coal resource than would be included in current economically recoverable reserve estimates. Where mining is no longer taking place, for economic or geological reasons, controlled gasification permits exploitation of the deposit (again a reaction of coal to form a synthesis gas) of coal seams in situ. The oxygen supply stage is one of the most expensive parts of any gasification project but the use of membranes is a potentially attractive approach for producing oxygen-enriched air. In this study, a variety of cost-effective membrane materials that gives an optimal amount of oxygen concentrations in the range of interest was designed and tested at diverse operating conditions. Oxygen-enriched atmosphere improves the combustion temperature but a decline is observed if oxygen concentration exceeds optimum. Experimental result also reveals the preparatory method, apparatus and performance of the fabricated membrane.Keywords: membranes, oxygen-enrichment, gasification, coal
Procedia PDF Downloads 460879 Conformational Switch of hRAGE upon Self-Association
Authors: Ikhlas Ahmed, Jamillah Zamoon
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The human receptor for advanced glycation end product is a plasma membrane receptor with an intrinsically disordered region. The protein consists of three extracellular domains, a single membrane spanning transmembrane domain, and a cytosolic domain which is intrinsically disordered and responsible for signaling. The disordered nature of the cytosolic domain allows it to be dynamic in solution. This receptor self-associates to higher forms. The association is triggered by ligand, metal or by the extracellular domain. Fluorescence spectroscopy technique is used to test the self-association of the different concentrations of the cytosolic domain. This work has concluded that the cytosolic domain of this receptor also self-associates. Moreover, the self-association does not require ligand or metal.Keywords: fluorescence spectroscopy, hRAGE, IDP, Self-association
Procedia PDF Downloads 361878 Extraction and Quantification of Triclosan in Wastewater Samples Using Molecularly Imprinted Membrane Adsorbent
Authors: Siyabonga Aubrey Mhlongo, Linda Lunga Sibali, Phumlane Selby Mdluli, Peter Papoh Ndibewu, Kholofelo Clifford Malematja
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This paper reports on the successful extraction and quantification of an antibacterial and antifungal agent present in some consumer products (Triclosan: C₁₂H₇Cl₃O₂)generally found in wastewater or effluents using molecularly imprinted membrane adsorbent (MIMs) followed by quantification and removal on a high-performance liquid chromatography (HPLC). Triclosan is an antibacterial and antifungal agent present in some consumer products like toothpaste, soaps, detergents, toys, and surgical cleaning treatments. The MIMs was fabricated usingpolyvinylidene fluoride (PVDF) polymer with selective micro composite particles known as molecularly imprinted polymers (MIPs)via a phase inversion by immersion precipitation technique. This resulted in an improved hydrophilicity and mechanical behaviour of the membranes. Wastewater samples were collected from the Umbogintwini Industrial Complex (UIC) (south coast of Durban, KwaZulu-Natal in South Africa). central UIC effluent treatment plant and pre-treated before analysis. Experimental parameters such as sample size, contact time, stirring speed were optimised. The resultant MIMs had an adsorption efficiency of 97% of TCS with reference to NIMs and bare membrane, which had 92%, 88%, respectively. The analytical method utilized in this review had limits of detection (LoD) and limits of quantification (LoQ) of 0.22, 0.71µgL-1 in wastewater effluent, respectively. The percentage recovery for the effluent samples was 68%. The detection of TCS was monitored for 10 consecutive days, where optimum TCS traces detected in the treated wastewater was 55.0μg/L inday 9 of the monitored days, while the lowest detected was 6.0μg/L. As the concentrations of analytefound in effluent water samples were not so diverse, this study suggested that MIMs could be the best potential adsorbent for the development and continuous progress in membrane technologyand environmental sciences, lending its capability to desalination.Keywords: molecularly imprinted membrane, triclosan, phase inversion, wastewater
Procedia PDF Downloads 123877 Nafion Nanofiber Mat in a Single Fuel Cell Test
Authors: Chijioke Okafor, Malik Maaza, Touhami Mokrani
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Proton exchange membrane, PEM was developed and tested for potential application in fuel cell. Nafion was electrospun to nanofiber network with the aid of poly(ethylene oxide), PEO, as a carrier polymer. The matrix polymer was crosslinked with Norland Optical Adhesive 63 under UV after compacting and annealing. The welded nanofiber mat was characterized for morphology, proton conductivity, and methanol permeability, then tested in a single cell test station. The results of the fabricated nanofiber membrane showed a proton conductivity of 0.1 S/cm at 25 oC and higher fiber volume fraction; methanol permeability of 3.6x10^-6 cm2/s and power density of 96.1 and 81.2 mW/cm2 for 5M and 1M methanol concentration respectively.Keywords: fuel cell, nafion, nanofiber, permeability
Procedia PDF Downloads 481876 Numerical Analysis of Real-Scale Polymer Electrolyte Fuel Cells with Cathode Metal Foam Design
Authors: Jaeseung Lee, Muhammad Faizan Chinannai, Mohamed Hassan Gundu, Hyunchul Ju
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In this paper, we numerically investigated the effect of metal foams on a real scale 242.57cm2 (19.1 cm × 12.7 cm) polymer electrolyte membrane fuel cell (PEFCs) using a three-dimensional two-phase PEFC model to substantiate design approach for PEFCs using metal foam as the flow distributor. The simulations were conducted under the practical low humidity hydrogen, and air gases conditions in order to observe the detailed operation result in the PEFCs using the serpentine flow channel in the anode and metal foam design in the cathode. The three-dimensional contours of flow distribution in the channel, current density distribution in the membrane and hydrogen and oxygen concentration distribution are provided. The simulation results revealed that the use of highly porous and permeable metal foam can be beneficial to achieve a more uniform current density distribution and better hydration in the membrane under low inlet humidity conditions. This study offers basic directions to design channel for optimal water management of PEFCs.Keywords: polymer electrolyte fuel cells, metal foam, real-scale, numerical model
Procedia PDF Downloads 239875 Ionic Polymer Actuators with Fast Response and High Power Density Based on Sulfonated Phthalocyanine/Sulfonated Polysulfone Composite Membrane
Authors: Taehoon Kwon, Hyeongrae Cho, Dirk Henkensmeier, Youngjong Kang, Chong Min Koo
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Ionic polymer actuators have been of interest in the bio-inspired artificial muscle devices. However, the relatively slow response and low power density were the obstacles for practical applications. In this study, ionic polymer actuators are fabricated with ionic polymer composite membranes based on sulfonated poly(arylene ether sulfone) (SPAES) and copper(II) phthalocyanine tetrasulfonic acid (CuPCSA). CuPCSA is an organic filler with very high ion exchange capacity (IEC, 4.5 mmol H+/g) that can be homogeneously dispersed on the molecular scale into the SPAES membrane. SPAES/CuPCSA actuators show larger ionic conductivity, mechanical properties, bending deformation, exceptional faster response to electrical stimuli, and larger mechanical power density (3028 W m–3) than Nafion actuators. This outstanding actuation performance of SPAES/CuPCSA composite membrane actuators makes them attractive for next generation transducers with high power density, which are currently developed biomimetic devices such as endoscopic surgery.Keywords: actuation performance, composite membranes, ionic polymer actuators, organic filler
Procedia PDF Downloads 278874 Experimental Study on Heat and Mass Transfer of Humidifier for Fuel Cell
Authors: You-Kai Jhang, Yang-Cheng Lu
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Major contributions of this study are threefold: designing a new model of planar-membrane humidifier for Proton Exchange Membrane Fuel Cell (PEMFC), an index to measure the Effectiveness (εT) of that humidifier, and an air compressor system to replicate related planar-membrane humidifier experiments. PEMFC as a kind of renewable energy has become more and more important in recent years due to its reliability and durability. To maintain the efficiency of the fuel cell, the membrane of PEMFC need to be controlled in a good hydration condition. How to maintain proper membrane humidity is one of the key issues to optimize PEMFC. We developed new humidifier to recycle water vapor from cathode air outlet so as to keep the moisture content of cathode air inlet in a PEMFC. By measuring parameters such as dry side air outlet dew point temperature, dry side air inlet temperature and humidity, wet side air inlet temperature and humidity, and differential pressure between dry side and wet side, we calculated indices obtained by dew point approach temperature (DPAT), water flux (J), water recovery ratio (WRR), effectiveness (εT), and differential pressure (ΔP). We discussed six topics including sealing effect, flow rate effect, flow direction effect, channel effect, temperature effect, and humidity effect by using these indices. Gas cylinders are used as sources of air supply in many studies of humidifiers. Gas cylinder depletes quickly during experiment at 1kW air flow rate, and it causes replication difficult. In order to ensure high stable air quality and better replication of experimental data, this study designs an air supply system to overcome this difficulty. The experimental result shows that the best rate of pressure loss of humidifier is 0.133×10³ Pa(g)/min at the torque of 25 (N.m). The best humidifier performance ranges from 30-40 (LPM) of air flow rates. The counter flow configured humidifies moisturizes the dry side inlet air more effectively than the parallel flow humidifier. From the performance measurements of the channel plates various rib widths studied in this study, it is found that the narrower the rib width is, the more the performance of humidifier improves. Raising channel width in same hydraulic diameter (Dh ) will obtain higher εT and lower ΔP. Moreover, increasing the dry side air inlet temperature or humidity will lead to lower εT. In addition, when the dry side air inlet temperature exceeds 50°C, the effect becomes even more obvious.Keywords: PEM fuel cell, water management, membrane humidifier, heat and mass transfer, humidifier performance
Procedia PDF Downloads 176873 Economic Analysis of a Carbon Abatement Technology
Authors: Hameed Rukayat Opeyemi, Pericles Pilidis Pagone Emmanuele, Agbadede Roupa, Allison Isaiah
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Climate change represents one of the single most challenging problems facing the world today. According to the National Oceanic and Administrative Association, Atmospheric temperature rose almost 25% since 1958, Artic sea ice has shrunk 40% since 1959 and global sea levels have risen more than 5.5cm since 1990. Power plants are the major culprits of GHG emission to the atmosphere. Several technologies have been proposed to reduce the amount of GHG emitted to the atmosphere from power plant, one of which is the less researched Advanced zero-emission power plant. The advanced zero emission power plants make use of mixed conductive membrane (MCM) reactor also known as oxygen transfer membrane (OTM) for oxygen transfer. The MCM employs membrane separation process. The membrane separation process was first introduced in 1899 when Walter Hermann Nernst investigated electric current between metals and solutions. He found that when a dense ceramic is heated, the current of oxygen molecules move through it. In the bid to curb the amount of GHG emitted to the atmosphere, the membrane separation process was applied to the field of power engineering in the low carbon cycle known as the Advanced zero emission power plant (AZEP cycle). The AZEP cycle was originally invented by Norsk Hydro, Norway and ABB Alstom power (now known as Demag Delaval Industrial turbomachinery AB), Sweden. The AZEP drew a lot of attention because its ability to capture ~100% CO2 and also boasts of about 30-50% cost reduction compared to other carbon abatement technologies, the penalty in efficiency is also not as much as its counterparts and crowns it with almost zero NOx emissions due to very low nitrogen concentrations in the working fluid. The advanced zero emission power plants differ from a conventional gas turbine in the sense that its combustor is substituted with the mixed conductive membrane (MCM-reactor). The MCM-reactor is made up of the combustor, low-temperature heat exchanger LTHX (referred to by some authors as air preheater the mixed conductive membrane responsible for oxygen transfer and the high-temperature heat exchanger and in some layouts, the bleed gas heat exchanger. Air is taken in by the compressor and compressed to a temperature of about 723 Kelvin and pressure of 2 Mega-Pascals. The membrane area needed for oxygen transfer is reduced by increasing the temperature of 90% of the air using the LTHX; the temperature is also increased to facilitate oxygen transfer through the membrane. The air stream enters the LTHX through the transition duct leading to inlet of the LTHX. The temperature of the air stream is then increased to about 1150 K depending on the design point specification of the plant and the efficiency of the heat exchanging system. The amount of oxygen transported through the membrane is directly proportional to the temperature of air going through the membrane. The AZEP cycle was developed using the Fortran software and economic analysis was conducted using excel and Matlab followed by optimization case study. The Simple bleed gas heat exchange layout (100 % CO2 capture), Bleed gas heat exchanger layout with flue gas turbine (100 % CO2 capture), Pre-expansion reheating layout (Sequential burning layout)–AZEP 85% (85% CO2 capture) and Pre-expansion reheating layout (Sequential burning layout) with flue gas turbine–AZEP 85% (85% CO2 capture). This paper discusses monte carlo risk analysis of four possible layouts of the AZEP cycle.Keywords: gas turbine, global warming, green house gas, fossil fuel power plants
Procedia PDF Downloads 397872 Surface Modification of SUS-304 Using Nitriding Treatment for Application of Bipolar Plates of Proton Exchange Membrane Fuel Cells
Authors: Wei-Ru Chang, Jenn-Jiang Hwang, Zen-Ting Hsiao, Shu-Feng Lee
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Proton exchange membrane (PEM) fuel cells are widely used in electrical systems as an economical, low-polluting energy source. This study investigates the effects of PEMFC gas nitriding treatment on metal bipolar plates. The test material was SUS304 stainless steel. The study explored five different pretreatment processes, varying the corrosion resistance and electrical conductivity conditions. The most effective process was industrial acid washing, followed by heating to 500 °C. Under the condition, the corrosion current density was 8.695 μA, significantly lower than that of the untreated pretreatment sample flakes, which was measured as 38.351 μA.Keywords: nitriding, bipolar, 304, corrosion, resistance, pretreatment
Procedia PDF Downloads 1087871 Structural and Functional Comparison of Untagged and Tagged EmrE Protein
Authors: S. Junaid S. Qazi, Denice C. Bay, Raymond Chew, Raymond J. Turner
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EmrE, a member of the small multidrug resistance protein family in bacteria is considered to be the archetypical member of its family. It confers host resistance to a wide variety of quaternary cation compounds (QCCs) driven by proton motive force. Generally, purification yield is a challenge in all membrane proteins because of the difficulties in their expression, isolation and solubilization. EmrE is extremely hydrophobic which make the purification yield challenging. We have purified EmrE protein using two different approaches: organic solvent membrane extraction and hexahistidine (his6) tagged Ni-affinity chromatographic methods. We have characterized changes present between ligand affinity of untagged and his6-tagged EmrE proteins in similar membrane mimetic environments using biophysical experimental techniques. Purified proteins were solubilized in a buffer containing n-dodecyl-β-D-maltopyranoside (DDM) and the conformations in the proteins were explored in the presence of four QCCs, methyl viologen (MV), ethidium bromide (EB), cetylpyridinium chloride (CTP) and tetraphenyl phosphonium (TPP). SDS-Tricine PAGE and dynamic light scattering (DLS) analysis revealed that the addition of QCCs did not induce higher multimeric forms of either proteins at all QCC:EmrE molar ratios examined under the solubilization conditions applied. QCC binding curves obtained from the Trp fluorescence quenching spectra, gave the values of dissociation constant (Kd) and maximum specific one-site binding (Bmax). Lower Bmax values to QCCs for his6-tagged EmrE shows that the binding sites remained unoccupied. This lower saturation suggests that the his6-tagged versions provide a conformation that prevents saturated binding. Our data demonstrate that tagging an integral membrane protein can significantly influence the protein.Keywords: small multidrug resistance (SMR) protein, EmrE, integral membrane protein folding, quaternary ammonium compounds (QAC), quaternary cation compounds (QCC), nickel affinity chromatography, hexahistidine (His6) tag
Procedia PDF Downloads 379