Search results for: membrane proteins

Authors: Yakup Ulusu, Numan Eczacıoğlu, İsa Gökçe, Helen Waller, Jeremy H. Lakey

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Besides having the appropriate amino acid sequence to perform the function of proteins, it is important to have correct conformation after this sequence to process. To consist of this conformation depends on the amino acid sequence at the primary structure, hydrophobic interaction, chaperones and enzymes in charge of folding etc. Misfolded proteins are not functional and tend to be aggregated. Cysteine originating disulfide cross-links make stable this conformation of functional proteins. When two of the cysteine amino acids come side by side, disulfide bond is established that forms a cystine bridge. Due to this feature cysteine plays an important role on the formation of three-dimensional structure of many proteins. There are two cysteine amino acids (C44, C69) in the Tol-A-III protein. Unlike protein disulfide bonds from within his own, any non-specific cystine bridge causes a change in the three dimensional structure of the protein. Proteins can be expressed in various host cells as directly or fusion (chimeric). As a result of overproduction of the recombinant proteins, aggregation of insoluble proteins in the host cell can occur by forming a crystal structure called inclusion body. In general fusion proteins are produced for provide affinity tags to make proteins more soluble and production of some toxic proteins via fusion protein expression system like pTolT. Proteins can be modified by using a site-directed mutagenesis. By this way, creation of non-specific disulfide crosslinks can be prevented at fusion protein expression system via the present cysteine replaced by another amino acid such as serine, glycine or etc. To do this, we need; a DNA molecule that contains the gene that encodes for the target protein, required primers for mutation to be designed according to site directed mutagenesis reaction. This study was aimed to be replaced cysteine encoding codon TGT with serine encoding codon AGT. For this sense and reverse primers designed (given below) and used site-directed mutagenesis reaction. Several new copy of the template plasmid DNA has been formed with above mentioned mutagenic primers via polymerase chain reaction (PCR). PCR product consists of both the master template DNA (wild type) and the new DNA sequences containing mutations. Dpn-l endonuclease restriction enzyme which is specific for methylated DNA and cuts them to the elimination of the master template DNA. E. coli cells obtained after transformation were incubated LB medium with antibiotic. After purification of plasmid DNA from E. coli, the presence of the mutation was determined by DNA sequence analysis. Developed this new plasmid is called PtolT-δ.

Keywords: site directed mutagenesis, Escherichia coli, pTolT, protein expression

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Authors: Manju Kanu, Subrata Sinha, Surabhi Johari

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Viral proteins of Ebola viruses belong to one of the best studied viruses; however no effective prevention against EBOV has been developed. Epitope-based vaccines provide a new strategy for prophylactic and therapeutic application of pathogen-specific immunity. A critical requirement of this strategy is the identification and selection of T-cell epitopes that act as vaccine targets. This study describes current methodologies for the selection process, with Ebola virus as a model system. Hence great challenge in the field of ebola virus research is to design universal vaccine. A combination of publicly available bioinformatics algorithms and computational tools are used to screen and select antigen sequences as potential T-cell epitopes of supertypes Human Leukocyte Antigen (HLA) alleles. MUSCLE and MOTIF tools were used to find out most conserved peptide sequences of viral proteins. Immunoinformatics tools were used for prediction of immunogenic peptides of viral proteins in zaire strains of Ebola virus. Putative epitopes for viral proteins (VP) were predicted from conserved peptide sequences of VP. Three tools NetCTL 1.2, BIMAS and Syfpeithi were used to predict the Class I putative epitopes while three tools, ProPred, IEDB-SMM-align and NetMHCII 2.2 were used to predict the Class II putative epitopes. B cell epitopes were predicted by BCPREDS 1.0. Immunogenic peptides were identified and selected manually by putative epitopes predicted from online tools individually for both MHC classes. Finally sequences of predicted peptides for both MHC classes were looked for common region which was selected as common immunogenic peptide. The immunogenic peptides were found for viral proteins of Ebola virus: epitopes FLESGAVKY, SSLAKHGEY. These predicted peptides could be promising candidates to be used as target for vaccine design.

Keywords: epitope, b cell, immunogenicity, ebola

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Authors: Jenny Radeva, Anke-Gundula Roth, Christian Goebbert, Robert Niestroj-Pahl, Lars Daehne, Axel Wolfram, Juergen Wiese

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Ceramic membranes for water purification combine excellent stability with long-life characteristics and high chemical resistance. Layer-by-Layer coating is a well-known technique for customization and optimization of filtration properties of membranes but is mostly used on polymeric membranes. Ceramic membranes comprising a metal oxide filtration layer of Al2O3 or TiO2 are charged and therefore highly suitable for polyelectrolyte adsorption. The high stability of the membrane support allows efficient backwash and chemical cleaning of the membrane. The presented study reports metal oxide/organic composite membrane with an increased rejection of bivalent salts like MgSO4 and the organic micropollutant Diclofenac. A self-build apparatus was used for applying the polyelectrolyte multilayers on the ceramic membrane. The device controls the flow and timing of the polyelectrolytes and washing solutions. As support for the Layer-by-Layer coat, ceramic mono-channel membranes were used with an inner capillary of 8 mm diameter, which is connected to the coating device. The inner wall of the capillary is coated subsequently with polycat- and anions. The filtration experiments were performed with a feed solution of MgSO4 and Diclofenac. The salt content of the permeate was detected conductometrically and Diclofenac was measured with UV-Adsorption. The concluded results show retention values of magnesium sulfate of 70% and diclofenac retention of 60%. Further experimental research studied various parameters of the composite membrane-like Molecular Weight Cut Off and pore size, Zeta potential and its mechanical and chemical robustness.

Keywords: water purification, polyelectrolytes, membrane modification, layer-by-layer coating, ceramic membranes

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Authors: Ziwei Wang, Andrea Lucotti, Luigi Brambilla, Matteo Tommasini, Chiara Bertarelli

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Surface-enhanced Raman Spectroscopy (SERS) is a highly sensitive detection that provides abundant information on low concentration analytes from various researching areas. Based on localized surface plasmon resonance, metal nanostructures including gold, silver and copper have been investigated as SERS substrate during recent decades. There has been increasing more attention of exploring good performance, homogenous, repeatable SERS substrates. Here, we show that electrospinning, which is an inexpensive technique to fabricate large-scale, self-standing and repeatable membranes, can be effectively used for producing SERS substrates. Nanoparticles and nanorods are added to the feed electrospinning solution to collect functionalized polymer fibrous mats. We report stable electrospun membranes as SERS substrate using gold nanorods (AuNRs) and poly(vinyl alcohol). Particularly, a post-processing crosslinking step using glutaraldehyde under acetone environment was carried out to the electrospun membrane. It allows for using the membrane in any liquid environment, including water, which is of interest both for sensing of contaminant in wastewater, as well as for biosensing. This crosslinked AuNRs/PVA membrane has demonstrated excellent performance as SERS substrate for low concentration 10-6 M Rhodamine 6G (Rh6G) aqueous solution. This post-processing for fabricating SERS substrate is the first time reported and proved through Raman imaging of excellent stability and outstanding performance. Finally, SERS tests have been applied to several analytes, and the application of AuNRs/PVA membrane is broadened by removing the detected analyte by rinsing. Therefore, this crosslinked AuNRs/PVA membrane is re-usable.

Keywords: SERS spectroscopy, electrospinning, crosslinking, composite materials

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Authors: Shayli Varasteh Moradi, Wayne A. Johnston, Dejan Gagoski, Kirill Alexandrov

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The demand for a rapid and more efficient technique to identify protein-protein interaction particularly in the areas of therapeutics and diagnostics development is growing. The method described here is a rapid in vitro protein-protein interaction analysis approach based on AlphaLISA technology combined with Leishmania tarentolae cell-free protein production (LTE) system. Cell-free protein synthesis allows the rapid production of recombinant proteins in a multiplexed format. Among available in vitro expression systems, LTE offers several advantages over other eukaryotic cell-free systems. It is based on a fast growing fermentable organism that is inexpensive in cultivation and lysate production. High integrity of proteins produced in this system and the ability to co-express multiple proteins makes it a desirable method for screening protein interactions. Following the translation of protein pairs in LTE system, the physical interaction between proteins of interests is analysed by AlphaLISA assay. The assay is performed using unpurified in vitro translation reaction and therefore can be readily multiplexed. This approach can be used in various research applications such as epitope mapping, antigen-antibody analysis and protein interaction network mapping. The intra-viral protein interaction network of Zika virus was studied using the developed technique. The viral proteins were co-expressed pair-wise in LTE and all possible interactions among viral proteins were tested using AlphaLISA. The assay resulted to the identification of 54 intra-viral protein-protein interactions from which 19 binary interactions were found to be novel. The presented technique provides a powerful tool for rapid analysis of protein-protein interaction with high sensitivity and throughput.

Keywords: AlphaLISA technology, cell-free protein expression, epitope mapping, Leishmania tarentolae, protein-protein interaction

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Authors: Wael El-Deeb

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The present study aimed to investigate the diagnostic importance of acute phase proteins in urinary tract infection (UTI) in cattle. We describe the clinical, bacteriological and biochemical findings in 99 lactating cows. Blood and urine samples from diseased (n=84) and control healthy cows (n=15) were submitted to laboratory investigations. The urine analysis revealed hematuria and pyuria in UTI group. The isolated bacteria were E.coli (43/84) Corynebacterium spp, (31/84), Proteus spp. (6/84) and Streptococcus spp (4/84). The concentrations of Haptoglobin (Hp), serum amyloid A (SAA), α1-Acid glycoprotein (AGP), fibrinogen (Fb), total protein, albumen, and globulin were higher in cows with UTI when compared to healthy ones. Fifty-one of 84 cows with UTI were successfully treated. The levels of Hp, SAA, AGP, total protein, and globulin were associated with the odds of treatment failure. Conclusively, acute phase proteins could be used as diagnostic and prognostic biomarkers in cows with UTI.

Keywords: cows, urinary, infections, haptoglobin, serum Amyloid A

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Authors: Arnab Mondal, Sanjeev Kumar Sharma, Ranjit Kumar Upadhyay

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We study the spatiotemporal dynamics of a neuronal cable. The processes of one- dimensional (1D) and 2D diffusion are considered for a single variable, which is the membrane voltage, i.e., membrane voltage diffusively interacts for spatiotemporal pattern formalism. The recovery and other variables interact through the membrane voltage. A 3D Leech-Heart (LH) model is introduced to investigate the nonlinear responses of an excitable neuronal cable. The deterministic LH model shows different types of firing properties. We explore the parameter space of the uncoupled LH model and based on the bifurcation diagram, considering v_k2_ashift as a bifurcation parameter, we analyze the 1D diffusion dynamics in three regimes: bursting, regular spiking, and a quiescent state. Depending on parameters, it is shown that the diffusive system may generate regular and irregular bursting or spiking behavior. Further, it is explored a 2D diffusion acting on the membrane voltage, where different types of patterns can be observed. The results show that the LH neurons with different firing characteristics depending on the control parameters participate in a collective behavior of an information processing system that depends on the overall network.

Keywords: bifurcation, pattern formation, spatio-temporal dynamics, stability analysis

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Authors: Jiahui Song

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The use of very high electric fields (~ 100kV/cm or higher) with pulse durations in the nanosecond range has been a recent development. The electric pulses have been used as tools to generate electroporation which has many biomedical applications. Most of the studies of electroporation have ignored possible thermal effects because of the small duration of the applied voltage pulses. However, it has been predicted membrane temperature gradients ranging from 0.2×109 to 109 K/m. This research focuses on thermal gradients that drives for electroporative enhancements, even though the actual temperature values might not have changed appreciably from their equilibrium levels. The dynamics of pore formation with the application of an externally applied electric field is studied on the basis of molecular dynamics (MD) simulations using the GROMACS package. Different temperatures are assigned to various regions to simulate the appropriate temperature gradients. The GROMACS provides the force fields for the lipid membranes, which is taken to comprise of dipalmitoyl-phosphatidyl-choline (DPPC) molecules. The water model mimicks the aqueous environment surrounding the membrane. Velocities of water and membrane molecules are generated randomly at each simulation run according to a Maxwellian distribution. For statistical significance, a total of eight MD simulations are carried out with different starting molecular velocities for each simulation. MD simulation shows no pore is formed in a 10-ns snapshot for a DPPC membrane set at a uniform temperature of 295 K after a 0.4 V/nm electric field is applied. A nano-sized pore is clearly seen in a 10-ns snapshot on the same geometry but with the top and bottom membrane surfaces kept at temperatures of 300 and 295 K, respectively. For the same applied electric field, the formation of nanopores is clearly demonstrated, but only in the presence of a temperature gradient. MD simulation results show enhanced electroporative effects arising from thermal gradients. The study suggests the temperature gradient is a secondary driver, with the electric field being the primary cause for electroporation.

Keywords: nanosecond, electroporation, thermal effects, molecular dynamics

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Authors: Min-Ah Woo, Min-Cheol Lim, Hyun-Joo Chang, Sung-Wook Choi

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We developed a paper-based colorimetric sensor utilizing magnetic nanoparticles conjugated with aptamers (MNP-Apts) against E. coli O157:H7. The MNP-Apts were applied to a test sample solution containing the target cells, and the solution was simply dropped onto PVDF (polyvinylidene difluoride) membrane. The membrane moves the sample radially to form the sample spots of different compounds as concentric rings, thus the MNP-Apts on the membrane enabled specific recognition of the target cells through a color ring generation by MNP-promoted colorimetric reaction of TMB (3,3',5,5'-tetramethylbenzidine) and H₂O₂. This method could be applied to rapidly and visually detect various bacterial pathogens in less than 1 h without cell culturing.

Keywords: aptamer, colorimetric sensor, E. coli O157:H7, magnetic nanoparticle, polyvinylidene difluoride

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

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

Keywords: green, microalgae, protein, solvents

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Authors: Narin Salehiyan, Ramin Ghasemi Shayan

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The largest genus of plant viruses, the potyvirus, is responsible for significant crop losses. Potyviruses are aphid sent in a nonpersistent way, and some of them are likewise seed communicated. As significant microorganisms, potyviruses are substantially more examined than other plant infections having a place with different genera, and their review covers numerous parts of plant virology, like utilitarian portrayal of viral proteins, sub-atomic communication with hosts and vectors, structure, scientific classification, development, the study of disease transmission, and determination. Biotechnological utilizations of potyviruses are likewise being investigated. During this last ten years, significant advances have been made in the comprehension of the sub-atomic science of these infections and the elements of their different proteins. Potyvirus multiplication, movement, and transmission, as well as potyvirus/plant compatible interactions, including pathogenicity and symptom determinants, are updated following a general overview of the family Potyviridae and the potyviral proteins. it end the survey giving data on biotechnological uses of potyviruses.

Keywords: virology, poty, virus, genome, genetic

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Authors: Maki Mizogami, Hironori Tsuchiya, Yoshiroh Hayabuchi, Kenji Shigemi

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Since drugs exhibit significant structure-dependent differences in activity and toxicity, their differentiation based on the mechanism of action should have implications for comparative drug efficacy and safety. We aimed to differentiate drug stereoisomers by their stereostructure-selective membrane interactions underlying pharmacological and toxicological effects. Biomimetic lipid bilayer membranes were prepared with phospholipids and sterols (either cholesterol or epicholesterol) to mimic the lipid compositions of neuronal and cardiomyocyte membranes and to provide these membranes with the chirality. The membrane preparations were treated with different classes of stereoisomers at clinically- and pharmacologically-relevant concentrations (25-200 μM), followed by measuring fluorescence polarization to determine the membrane interactivity of drugs to change the physicochemical property of membranes. All the tested drugs acted on lipid bilayers to increase or decrease the membrane fluidity. Drug stereoisomers could not be differentiated when interacting with the membranes consisting of phospholipids alone. However, they stereostructure-selectively interacted with neuro-mimetic and cardio-mimetic membranes containing 40 mol% cholesterol ((3β)-cholest-5-en-3-ol) to show the relative potencies being local anesthetic R(+)-bupivacaine > rac-bupivacaine > S(‒)-bupivacaine, α2-adrenergic agonistic D-medetomidine > rac-medetomidine > L-medetomidine, β-adrenergic antagonistic R(+)-propranolol > rac-propranolol > S(–)-propranolol, NMDA receptor antagonistic S(+)-ketamine > rac-ketamine, analgesic monoterpenoid (+)-menthol > (‒)-menthol, non-steroidal anti-inflammatory S(+)-ibuprofen > rac-ibuprofen > R(‒)-ibuprofen, and bioactive flavonoid (+)-epicatechin > (‒)-epicatechin. All of the order of membrane interactivity were correlated to those of beneficial and adverse effects of the tested stereoisomers. In contrast, the membranes prepared with epicholesterol ((3α)-chotest-5-en-3-ol), an epimeric form of cholesterol, reversed the rank order of membrane interactivity to be S(‒)-enantiomeric > racemic > R(+)-enantiomeric bupivacaine, L-enantiomeric > racemic > D-enantiomeric medetomidine, S(–)-enantiomeric > racemic > R(+)-enantiomeric propranolol, racemic > S(+)-enantiomeric ketamine, (‒)-enantiomeric > (+)-enantiomeric menthol, R(‒)-enantiomeric > racemic > S(+)-enantiomeric ibuprofen, and (‒)-enantiomeric > (+)-enantiomeric epicatechin. The opposite configuration allows drug molecules to interact with chiral sterol membranes enantiomer-selectively. From the comparative results, it is speculated that a 3β-hydroxyl group in cholesterol is responsible for the enantioselective interactions of drugs. In conclusion, the differentiation of drug stereoisomers by their stereostructure-selective membrane interactions would be useful for designing and predicting drugs with higher activity and/or lower toxicity.

Keywords: chiral membrane, differentiation, drug stereoisomer, enantioselective membrane interaction

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Authors: Elif Tugce Aksun Tumerkan, Chris Lowe, Hannah Krupa

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Fluorescent proteins are self-sufficient in forming chromophores with a visible wavelength from 3 amino acids sequence within their own polypeptide structure. This chromophore – a molecule that absorbs a photon of light and exhibits an energy transition equal to the energy of the absorbed photon. Fluorescent proteins (FPs) consisted of a chain of 238 amino acid residues and composed of 11 beta strands shaped in a cylinder surrounding an alpha helix structure. A better understanding of the system of the chromospheres and the increasing advance in protein engineering in recent years, the properties of FPs offers the potential for new applications. They have used sensors and probes in molecular biology and cell-based research that giving a chance to observe these FPs tagged cell localization, structural variation and movement. For clarifying functional uses of fluorescent proteins, electrophoretic properties of these proteins are one of the most important parameters. Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) analysis is used for determining electrophoretic properties commonly. While there are many techniques are used for determining the functionality of protein-based research, SDS-PAGE analysis can only provide a molecular level assessment of the proteolytic fragments. Before SDS-PAGE analysis, fluorescent proteins need to successfully purified. Due to directly purification of the target, FPs is difficult from the animal, gene expression is commonly used which must be done by transformation with the plasmid. Furthermore, used gel within electrophoresis and staining agents properties have a key role. In this review, the different factors that have the impact on the electrophoretic properties of fluorescent proteins explored. Fluorescent protein separation and purification are the essential steps before electrophoresis that should be done very carefully. For protein purification, gene expression process and following steps have a significant function. For successful gene expression, the properties of selected bacteria for expression, used plasmid are essential. Each bacteria has own characteristics which are very sensitive to gene expression, also used procedure is the important factor for fluorescent protein expression. Another important factors are gel formula and used staining agents. Gel formula has an effect on the specific proteins mobilization and staining with correct agents is a key step for visualization of electrophoretic bands of protein. Visuality of proteins can be changed depending on staining reagents. Apparently, this review has emphasized that gene expression and purification have a stronger effect than electrophoresis protocol and staining agents.

Keywords: cell biology, gene expression, staining agents, SDS-page

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Authors: Prakash Thapa, Gye Choon Park, Sung Gi Kwon, Jin Lee

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The generation of electric power from a proton exchange membrane (PEM) fuel cell is influenced by temperature, pressure, humidity, flow rate of reactant gaseous and partial flooding of membrane electrode assembly (MEA). Among these factors, temperature and cathode flooding are the most affecting parameters on the performance of fuel cell. This paper describes the detail design and effect of these parameters on PEM fuel cell. Performance of all parameters was monitored, analyzed and controlled by using 5KWatt PEM fuel cell. In the real-time data communication for remote monitoring and control of PEM fuel cell, a normalized least mean square algorithm in cognitive radio environment is used. By the use of this method, probability of energy signal detection will be maximum which solved the frequency shortage problem. So the monitoring system hanging out and slow speed problem will be solved. Also from the control unit, all parameters are controlled as per the system requirement. As a result, PEM fuel cell generates maximum electricity with better performance.

Keywords: proton exchange membrane (PEM) fuel cell, pressure, temperature and humidity sensor (PTH), efficiency curve, cognitive radio network (CRN)

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Authors: G. Carrero, C. Contreras, M. J. Hendzel

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Linker histones or histones H1 are highly mobile nuclear proteins that regulate the organization of chromatin and limit DNA accessibility by binding to the chromatin structure (DNA and associated proteins). It is known that this binding process is driven by both slow (strong binding) and rapid (weak binding) interactions. However, the exact binding mechanism has not been fully described. Moreover, the existing models only account for one type of bound population that does not distinguish explicitly between the weakly and strongly bound proteins. Thus, we propose different systems of reaction-diffusion equations to describe explicitly the rapid and slow interactions during a FRAP (Fluorescence Recovery After Photobleaching) experiment. We perform a model comparison analysis to characterize the binding mechanism of histone H1 and provide new meaningful biophysical information on the kinetics of histone H1.

Keywords: FRAP (Fluorescence Recovery After Photobleaching), histone H1, histone H1 binding kinetics, linker histone, reaction-diffusion equation

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Authors: Mahsa Jalili, Trude Johansen, Signe Dille Lovmo, Turid Rustad, Rolf Erik Olsen, Atle M. Bones

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The quality of fish muscle is a key factor in fish industry, and dietary ingredients can influence fish quality. The aim of this study was to examine the impact of krill meal, soybean meal, Bactocell® and butyrate fortified feeds and control diet on characteristics of salmon fillet. Thirty Atlantic salmon (6 per each group) were farmed for 12 weeks. All the fish were killed and frozen immediately. The white muscle from top posterior part of dorsal fin was dissected to analyze fat content, carotenoid content, content of water-soluble and salt-soluble proteins, cathepsin B and cathepsin B-L activities. ANOVA test was used to analyze mean and standard error of mean values at 0.05 significance level. There were significant difference in cathepsin B activity, water-soluble proteins and salt-soluble proteins (p-value= 0.005, 0.009 and 0.002). The mean values of other factors were not significantly different among the groups. Cathepsin B activity was higher in soymeal group. Water-soluble proteins were reported higher in soy meal and krill groups and salt-soluble proteins were significantly higher in soy meal and butyrate rich diets. Although soy meal has proven effect on enteritis, it results in higher percentage of protein in fillets. On the other hand, this feeding may have role in textural deterioration of fillets owing to higher values of endogenous cathepsin B in soymeal group.

Keywords: aquaculture, food quality, Krill protein extract, prebiotics, probiotics, Salmo salar, soy

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Authors: Nilubon Thongtan, Wilai Chiemchaisri, Chart Chiemchaisri

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Purpose of study is to investigate performance of two-stage membrane bioreactor (2S-MBR) to treat trimethoprim and sulfamethoxazole in solid waste leachate. This system consists of 2 tanks, anoxic tank with incline plates and MBR tank. The system was operated at 12 h-HRT each, of which the MBR MLSS concentration was operated at 25,000-35,000 mg/L. The average sCOD concentration of the fed leachate was 6,310±3,595 mg/L. It shows that high organic removals in terms of sCOD and sBOD were achieved as of 97-99% and 99%, respectively. The TKN and NH3-N removals were 76-98% and 91-99%, respectively. Concurrently, trimethoprim and sulfamethoxazole were detected in the leachate with concentrations of 113-0 μg/L and 74-2 μg/L, respectively. High removals of trimethoprim and sulfamethoxazole were also found as of 95-99% and 85-95%, respectively. In sum, this MBR feature and operation gave achievement in treatment of macro-pollutants including trimethoprim and sulfamethoxazole existing in low levels in the solid waste leachate.

Keywords: membrane bioreactor, solid waste leachate, sulfamethoxazole, trimethoprim

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Authors: Ahmad Kayvani Fard, Yehia Manawi

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Qatar’s primary source of fresh water is through seawater desalination. Amongst the major processes that are commercially available on the market, the most common large scale techniques are Multi-Stage Flash distillation (MSF), Multi Effect distillation (MED), and Reverse Osmosis (RO). Although commonly used, these three processes are highly expensive down to high energy input requirements and high operating costs allied with maintenance and stress induced on the systems in harsh alkaline media. Beside that cost, environmental footprint of these desalination techniques are significant; from damaging marine eco-system, to huge land use, to discharge of tons of GHG and huge carbon footprint. Other less energy consuming techniques based on membrane separation are being sought to reduce both the carbon footprint and operating costs is membrane distillation (MD). Emerged in 1960s, MD is an alternative technology for water desalination attracting more attention since 1980s. MD process involves the evaporation of a hot feed, typically below boiling point of brine at standard conditions, by creating a water vapor pressure difference across the porous, hydrophobic membrane. Main advantages of MD compared to other commercially available technologies (MSF and MED) and specially RO are reduction of membrane and module stress due to absence of trans-membrane pressure, less impact of contaminant fouling on distillate due to transfer of only water vapor, utilization of low grade or waste heat from oil and gas industries to heat up the feed up to required temperature difference across the membrane, superior water quality, and relatively lower capital and operating cost. To achieve the objective of this study, state of the art flat-sheet cross-flow DCMD bench scale unit was designed, commissioned, and tested. The objective of this study is to analyze the characteristics and morphology of the membrane suitable for DCMD through SEM imaging and contact angle measurement and to study the water quality of distillate produced by DCMD bench scale unit. Comparison with available literature data is undertaken where appropriate and laboratory data is used to compare a DCMD distillate quality with that of other desalination techniques and standards. Membrane SEM analysis showed that the PTFE membrane used for the study has contact angle of 127º with highly porous surface supported with less porous and bigger pore size PP membrane. Study on the effect of feed solution (salinity) and temperature on water quality of distillate produced from ICP and IC analysis showed that with any salinity and different feed temperature (up to 70ºC) the electric conductivity of distillate is less than 5 μS/cm with 99.99% salt rejection and proved to be feasible and effective process capable of consistently producing high quality distillate from very high feed salinity solution (i.e. 100000 mg/L TDS) even with substantial quality difference compared to other desalination methods such as RO and MSF.

Keywords: membrane distillation, waste heat, seawater desalination, membrane, freshwater, direct contact membrane distillation

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Authors: M. P. S. Tomar, Neelam Bansal

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Histomorphological, histochemical and scanning electron microscopic observations were recorded in developing cornea of buffalo fetuses. The samples from fetal cornea were collected in appropriate fixative from slaughter house and Veterinary Clinics, GADVASU, Ludhiana. The microscopic slides were stained for detailed histomorphological and histochemical studies. The scanning electron microscopic studies were performed at Electron microscopy & Nanobiology Lab, PAU Ludhiana. In present study, it was observed that, in 36 days (d) fetus, the corneal epithelium was well marked single layered structure which was placed on stroma mesenchyme. Cornea appeared as the continuation of developing sclera. The thickness of cornea and its epithelium increased as well as the epithelium started becoming double layered in 47d fetus at corneo-scleral junction. The corneal thickness in this stage suddenly increased thus easily distinguished from developing sclera. The separation of corneal endothelium from stroma was evident as a single layered epithelium. The stroma possessed numerous fibroblasts in 49d stage eye. Descemet’s membrane was appeared at 52d stage. The limbus area was separated by a depression from the developing cornea in 61d stage. In 65d stage, the Bowman’s layer was more developed. Fibroblasts were arranged parallel to each other as well as parallel to the surface of developing cornea in superficial layers. These fibroblasts and fibers were arranged in wavy pattern in the center of stroma. Corneal epithelium started to be stratified as a double layered epithelium was present in this age of fetal eye. In group II (>120 Days), the corneal epithelium was stratified towards a well marked irido-corneal angle. The stromal fibroblasts followed a complete parallel arrangement in its entire thickness. In full term fetuses, a well developed cornea was observed. It was a fibrous layer which had five distinct layers. From outside to inwards were described as the outer most layer was the 7-8 layered corneal epithelial, subepithelial basement membrane (Bowman’s membrane), substantia propria or stroma, posterior limiting membrane (Descemet’s membrane) and the posterior epithelium (corneal endothelium). The corneal thickness and connective tissue elements were continued to be increased. It was 121.39 + 3.73µ at 36d stage which increased to 518.47 + 4.98 µ in group III fetuses. In fetal life, the basement membrane of corneal epithelium and endothelium depicted strong to intense periodic Acid Schiff’s (PAS) reaction. At the irido-corneal angle, the endothelium of blood vessels was also positive for PAS activity. However, cornea was found mild positive for alcian blue reaction. The developing cornea showed strong reaction for basic proteins in outer epithelium and the inner endothelium layers. Under low magnification scanning electron microscope, cornea showed two types of cells viz. light cells and dark cells. The light cells were smaller in size and had less number of microvilli in their surface than in the dark cells. Despite these surface differences between light and dark cells, the corneal surface showed the same general pattern of microvilli studding all exposed surfaces out to the cell margin. which were long (with variable height), slight tortuous slender and possessed a micro villus shaft with a very prominent knob.

Keywords: buffalo, cornea, eye, fetus, ontogeny, scanning electron microscopy

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Authors: Netsanet Kebede Hundessa

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Substrate modification of thin film composite (TFC) membranes with various crosslinkers is typically necessary for organic solvent nanofiltration (OSN) applications. This modification is aimed at enhancing membrane stability and solvent resistance, but it often results in a decline in permeance. This study introduces a distinct approach by developing a coordination-crosslinked polyimide substrate, which differs from the covalently-crosslinked substrates traditionally used. This developed substrate achieves enhanced solvent resistance, improved hydrophilicity, and optimized porous microstructure simultaneously. The study investigates the effects of an alkaline coagulation bath, subsequent ion exchange, and further solvent activation. The resulting TFC membrane successfully overcomes the typical permeability-selectivity trade-off of OSN membranes. It demonstrates significantly improved solvent permeance (1.5–2 times higher than previously reported data) with values of 65.2 LMH/bar for methanol, 33.1 LMH/bar for ethanol, and 59.1 LMH/bar for acetone while maintaining competitive solute rejection (>98% for Rose Bengal). This research is expected to provide a new direction for developing high-performance OSN composite membranes and other separation applications.

Keywords: metal coordinatiom, thin film composite membrane, organic solvent nanofiltration, solvent activation

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Authors: Y. H. Hwang, J. Won, Y. S. Kang

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Since ionic liquids have a special interaction with gas specially CO2 and/or olefin, supported ionic liquids membrane (SILM) are fabricated for practical gas separation. However, SILM has a problem in practical application due to the low mechanical stability under high pressure for gas separation. In order to improve the mechanical strength of the selective ionic liquid layer, we prepared supported ion gel membrane by the formation of gel on the surface of Nylon support. The ion gel was prepared by the addition of poly(styrene-block-ethyleneoxide-block-styrene) triblock copolymer in four tricyanomethanide ionic liquids have different cation; 1-ethyl-3-methlyimidazolium tricyanomethanide, 1-butyl-3-methlyimidazolium tricyanomethanide, 1-butyl-1-methylpyrrolidinium tricyanomethanide, 1-butyl-4-methylpyridinium tricyanomethanide using methylenechloride as a solvent. The characters of ion gel with different cation were studied. Four different gases (CO2, N2, O2, and CH4) permeance were measured at room temperature by bubble flow meter and cation effect of tricyanomethanide ionic liquids investigated.

Keywords: membrane, ionic liquid, ion gel, nanostructure

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Authors: Nasser A. M. Barakat

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Membrane technology is the most promising process for oil-water separation operation if the hydrophilicity, fouling and reusability properties could be improved. In this study, novel effective and reusable membrane for oil-water separation process is introduced based on modification of polysulfone (PSF) electrospun nanofiber mats. The modification process was achieved by incorporation of NaOH nanoparticles inside the PSF nanofibers, and formation of a thin layer from a polyamide polymer on the surface of the electrospun mat. Typically, solutions composed of PSF and NaOH (twelve solutions were prepared based on different PSF concentrations; 15, 18 and 20 wt%, and various NaOH content; 1.5, 1.7 and 2.5 wt%) have been electrospun, then the dried nanofiber mats were treated by m-phenylenediamine and 1,3,5-benzenetricarbonyl chloride to form polyamide thin layer on the surface of the mats. The results indicated that incorporation of NaOH and the formed polyamide could decrease the water contact angle from ~ 130˚ to 13˚ for the nanofiber mats obtained from 20 wt% PSF solutions containing 1.7 wt% sodium hydroxide powders. Interestingly, the membrane having the lowest contact angle could separate oil-water mixture for three successive cycles and 100% removal of the oil with relatively high water flux; 5.5 m3/m2.day. Overall, simplicity of the manufacturing technique, and effectiveness and reusability of the produced nanofiber mats open new avenue for the introduced as promising membranes for the oil-water separation process.

Keywords: electrospinning, oil-water separation, hydrophilic membrane, nanofibers

Procedia PDF Downloads 341

Authors: Elif Tugce Aksun Tumerkan

Abstract:

Fluorescent proteins (FPs) have become very popular since green fluorescent protein discovered from crystal jellyfish. It is known that Anthozoa species have a wide range of chromophore organisms, and the initial crystal structure for non-fluorescent chromophores obtained from the reef-building coral has been determined. There are also differently coloured pigments in non-bioluminescent Anthozoa zooxanthellate and azooxanthellate which are frequently members of the GFP-like protein family. The development of fluorescent proteins (FPs) and their applications is an outstanding example of basic science leading to practical biotechnological and medical applications. Fluorescent proteins have several applications in science and are used as important indicators in molecular biology and cell-based research. With rising interest in cell biology, FPs have used as biosensor indicators and probes in pharmacology and cell biology. Using fluorescent proteins in genetically encoded metabolite sensors has many advantages than chemical probes for metabolites such as easily introduced into any cell or organism in any sub-cellular localization and giving chance to fixing to fluoresce of different colours or characteristics. There are different factors effects to signalling mechanism when they used as a biosensor. While there are wide ranges of research have been done on the significance and applications of fluorescent proteins, the cell signalling response of FPs and target cell are less well understood. In this study, it was aimed to clarify the response of adaptive mechanisms of coral species such as pH, temperature and symbiotic relationship and target cells properties on the signalling capacity. Corals are a rich natural source of fluorescent proteins that change with environmental conditions such as light, heat stress and injury. Adaptation mechanism of coral species to these types of environmental variations is important factor due to FPs properties have affected by this mechanism. Since fluorescent proteins obtained from nature, their own ecological property like the symbiotic relationship is observed very commonly in coral species and living conditions have the impact on FPs efficiency. Target cell properties also have an effect on signalling and visualization. The dynamicity of detector that used for reading fluorescence and the level of background fluorescence are key parameters for the quality of the fluorescent signal. Among the factors, it can be concluded that coral species adaptive characteristics have the strongest effect on FPs signalling capacity.

Keywords: biosensor, cell biology, environmental conditions, fluorescent protein, sea anemone

Procedia PDF Downloads 169

Authors: R. Gounina-Allouane, N. Ouali, F. Z. Berrabah, A. Bentaleb

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For a long time, the Gram-positive spore-forming bacteria Bacillus thuringiensis (Bt) has been widely used in biological control against devastating and disease vectors insects. This is due to the insecticidal activity of its crystalline parasporal inclusion (crystals) predominantly comprised of one or more proteins (Cry and Cyt proteins) also called δ-endotoxins, produced during sporulation. The shape and composition of Bt crystals vary among strains and crystalline proteins are extremely varied (more than 475 cry gene were discovered). The insecticidal activity of Bt crystals is very well studied, thus their insecticidal mode of action is well established, however, their antimicrobial effect is largely unknown. The lack of data on the antimicrobial effect of crystalline proteins of Bt and the need for searching new antimicrobial molecules encouraged us to carried out this study. The antibacterial effect of δ-endotoxines produced by two Bt stains; a strain isolated from soil at northern of Algeria (Bt 7.2.B), and a strain isolated from a bioinsecticide (Bacillus thuringiensis var aizawai), activated by proteolysis, was assayed on clinical bacterial strains and ATCC collection ones respectively. Gram positive and negative clinical bacterial strains (Escherichia coli, Klebsiella pneumonaie, Pseudomonas aeruginosa, Staphylococcus aureus) were sensitive to activated Bt 72B endotoxins. Similarly, bacterial strains from ATCC collection (Escherichia coli ATCC 25922, Pseudomonas aerugenosa ATCC 27853, Staphylococcus aureus ATCC 25923) were sensitive to activated B. thuringiensis var aizawai δ-endotoxines. The activated δ-endotoxins were separated by SDS-PAGE.

Keywords: Bacillus thuringiensis, crystals, cry proteins, δ-endotoxins, antibacterial activity

Procedia PDF Downloads 448

Authors: M. Kumpugdee-Vollrath, T. G. D. Phu, M. Helmis

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A suitable model membrane to study the pharmacological effect of pharmaceutical products is human stratum corneum because this layer of human skin is the outermost layer and it is an important barrier to be passed through. Other model membranes which were also used are for example skins from pig, mouse, reptile or fish. We are interested in fish skins in this project. The advantages of the fish skins are, that they can be obtained from the supermarket or fish shop. However, the fish skins should be freshly prepared and used directly without storage. In order to understand the effect of different model drugs e.g. lidocaine HCl, resveratrol, paracetamol, ibuprofen, acetyl salicylic acid on the properties of the model membrane from various types of fishes e.g. trout, salmon, cod, plaice permeation tests were performed and differential scanning calorimetry was applied.

Keywords: fish skin, model membrane, permeation, DSC, lidocaine HCl, resveratrol, paracetamol, ibuprofen, acetyl salicylic acid

Procedia PDF Downloads 470

Authors: Ming-Yeh Lu, Shiao-Shing Chen, Saikat Sinha Ray, Hung-Te Hsu

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Recently, anaerobic membrane bioreactors (AnMBRs) has been widely utilized, which combines anaerobic biological treatment process and membrane filtration, that can be present an attractive option for wastewater treatment and water reuse. Conventional AnMBR is having several advantages, such as improving effluent quality, compact space usage, lower sludge yield, without aeration and production of energy. However, the removal of nitrogen and phosphorus in the AnMBR permeate was negligible which become the biggest disadvantage. In recent years, forward osmosis (FO) is an emerging technology that utilizes osmotic pressure as driving force to extract clean water without additional external pressure. The pore size of FO membrane is kindly mentioned the pore size, so nitrogen or phosphorus could effectively improve removal of nitrogen or phosphorus. Anaerobic bioreactor with FO membrane (AnOMBR) can retain the concentrate organic matters and nutrients. However, phosphorus is a non-renewable resource. Due to the high rejection property of FO membrane, the high amount of phosphorus could be recovered from the combination of AnMBR and FO. In this study, development of novel submerged anaerobic osmotic membrane bioreactor integrated with periodic microfiltration (MF) extraction for simultaneous phosphorus and clean water recovery from wastewater was evaluated. A laboratory-scale AnOMBR utilizes cellulose triacetate (CTA) membranes with effective membrane area of 130 cm² was fully submerged into a 5.5 L bioreactor at 30-35℃. Active layer-facing feed stream orientation was utilized, for minimizing fouling and scaling. Additionally, a peristaltic pump was used to circulate draw solution (DS) at a cross flow velocity of 0.7 cm/s. Magnesium sulphate (MgSO₄) solution was used as DS. Microfiltration membrane periodically extracted about 1 L solution when the TDS reaches to 5 g/L to recover phosphorus and simultaneous control the salt accumulation in the bioreactor. During experiment progressed, the average water flux was achieved around 1.6 LMH. The AnOMBR process show greater than 95% removal of soluble chemical oxygen demand (sCOD), nearly 100% of total phosphorous whereas only partial removal of ammonia, and finally average methane production of 0.22 L/g sCOD was obtained. Therefore, AnOMBR system periodically utilizes MF membrane extracted for phosphorus recovery with simultaneous pH adjustment. The overall performance demonstrates that a novel submerged AnOMBR system is having potential for simultaneous wastewater treatment and resource recovery from wastewater, and hence, the new concept of this system can be used to replace for conventional AnMBR in the future.

Keywords: anaerobic treatment, forward osmosis, phosphorus recovery, membrane bioreactor

Procedia PDF Downloads 270

Authors: S. Oluwagbemiga Alayande, E. Olugbenga Dare, Titus A. M. Msagati, A. Kehinde Akinlabi , P. O. Aiyedun

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This paper presents a cheap route procedure for the preparation of a potential oil membrane with superhydrophobic /superoleophillic properties for selective removal of crude oil from water. In these study, expanded polystyrene (EPS) was electrospun to produce beaded fibers in which zeolite was introduced to the polymer matrix in order to impart rough surface to non-beaded fiber. Films of the EPS and EPS/Zeolite solutions were also made for comparative study. The electrospun fibers EPS, EPS/Zeolite and resultant films were characterized using SEM, BET, FTIR and optical contact angle. The fibers exhibited superhydrophic and superoleophillic wetting properties with water and crude oil. The selective removal of crude oil presents new opportunity for the re-use of EPS as adsorbent in petroleum/petrochemical industry.

Keywords: expanded polystyrene, superhydrophobic, superoleophillic, oil-membrane

Procedia PDF Downloads 472

Authors: Lozano Paul

Abstract:

Coupled Membrane Separation Technology (CMST), including Cross Flow Microfiltration (CFM) and Reverse Osmosis (RO), are used to concentrate microalgae biomass or/and to extract and concentrate water-soluble metabolites produced during micro-algae production cycle, as well as water recycling. Micro-algae biomass was produced using different feeding mixtures of ingredients: pure chemical origin compounds and natural/ecological water-extracted components from available local plants. Micro-algae was grown either in conventional plastic bags (100L/unit) or in small-scale innovative bioreactors (75L). Biomass was concentrated as CFM retentate using a P19-60 ceramic membrane (0.2μm pore size), and water-soluble micro-algae metabolites left in the CFM filtrate were concentrated by RO. Large volumes of water (micro-algae culture media) of were recycled by the CMTS for another biomass production cycle.

Keywords: extraction, membrane process, microalgae, natural compound

Procedia PDF Downloads 279

Authors: Ayat N. El-Shazly, Dina Magdy Abdo, Hamdy Maamoun Abdel-Ghafar, Xiangju Song, Heqing Jiang

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Product water recovery and draw solution (DS) reuse is the most energy-intensive stage in forwarding osmosis (FO) technology. Sucrose solution is the most suitable DS for FO application in food and beverages. However, sucrose DS recovery by conventional pressure-driven or thermal-driven concentration techniques consumes high energy. Herein, we developed a spontaneous and sustainable solar-driven evaporation process based on a photothermal membrane for the concentration and recovery of sucrose solution. The photothermal membrane is composed of multi-walled carbon nanotubes (f-MWCNTs)photothermal layer on a hydrophilic polyvinylidene fluoride (PVDF) substrate. The f-MWCNTs photothermal layer with a rough surface and interconnected network structures not only improves the light-harvesting and light-to-heat conversion performance but also facilitates the transport of water molecules. The hydrophilic PVDF substrate can promote the rapid transport of water for adequate water supply to the photothermal layer. As a result, the optimized f-MWCNTs/PVDF photothermal membrane exhibits an excellent light absorption of 95%, and a high surface temperature of 74 °C at 1 kW m−2 . Besides, it realizes an evaporation rate of 1.17 kg m−2 h−1 for 5% (w/v) of sucrose solution, which is about 5 times higher than that of the natural evaporation. The designed photothermal evaporation process is capable of concentrating sucrose solution efficiently from 5% to 75% (w/v), which has great potential in FO process and juice concentration.

Keywords: solar, pothothermal, membrane, MWCNT

Procedia PDF Downloads 99

Authors: Vadim V. Zefirov, Victor E. Sizov, Marina A. Pigaleva, Igor V. Elmanovich, Mikhail S. Kondratenko, Marat O. Gallyamov

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This work presents a novel method for treating porous hydrophobic polyolefin membranes using supercritical carbon dioxide that allows usage of the modified membrane in redox flow batteries with an aqueous electrolyte. Polyolefin membranes are well known and widely used, however, they cannot be used as separators in redox flow batteries with an aqueous electrolyte since they have insufficient wettability, and therefore do not provide sufficient proton conductivity. The main aim of the presented work was the development of hydrophilic composites based on cheap membranes and precursors. Supercritical fluid was used as a medium for the deposition of the hydrophilic phase on the hydrophobic surface of the membrane. Due to the absence of negative capillary effects in a supercritical medium, a homogeneous composite is obtained as a result of synthesis. The in-situ synthesized silicon oxide nanoparticles and the chitosan polymer layer act as the hydrophilic phase and not only increase the affinity of the membrane towards the electrolyte, but also reduce the pore size of the polymer matrix, which positively affects the ion selectivity of the membrane. The composite material obtained as a result of synthesis has enhanced hydrophilic properties and is capable of providing proton conductivity in redox flow batteries. The morphology of the obtained composites was characterized by electron microscopy. To analyze the phase composition, infrared spectroscopy was used. The hydrophilic properties were studied by water contact angle measurements. In addition, the proton conductivity and ion selectivity of the obtained samples were studied, and tests in real redox flow batteries were performed. As a result, modified membrane was characterised in detail and moreover it was shown that modified cheap polyolefin membranes have pronounced proton conductivity and high ion selectivity, so their performance in a real redox flow battery approaches expensive commercial analogues, reaching 70% of energy efficiency.

Keywords: carbon dioxide, chitosan, polymer membrane, redox flow batteries, silica nanoparticles, supercritical fluid

Procedia PDF Downloads 153