Search results for: ceramic nanoporous membrane
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 1471

Search results for: ceramic nanoporous membrane

691 Core-Shell Nanofibers for Prevention of Postsurgical Adhesion

Authors: Jyh-Ping Chen, Chia-Lin Sheu

Abstract:

In this study, we propose to use electrospinning to fabricate porous nanofibrous membranes as postsurgical anti-adhesion barriers and to improve the properties of current post-surgical anti-adhesion products. We propose to combine FDA-approved biomaterials with anti-adhesion properties, polycaprolactone (PCL), polyethylene glycol (PEG), hyaluronic acid (HA) with silver nanoparticles (Ag) and ibuprofen (IBU), to produce anti-adhesion barrier nanofibrous membranes. For this purpose, PEG/PCL/Ag/HA/IBU core-shell nanofibers were prepared. The shell layer contains PEG + PCL to provide mechanical supports and Ag was added to the outer PEG-PCL shell layer during electrospinning to endow the nanofibrous membrane with anti-bacterial properties. The core contains HA to exert anti-adhesion and IBU to exert anti-inflammation effects, respectively. The nanofibrous structure of the membranes can reduce cell penetration while allowing nutrient and waste transports to prevent postsurgical adhesion. Nanofibers with different core/shell thickness ratio were prepared. The nanofibrous membranes were first characterized for their physico-chemical properties in detail, followed by in vitro cell culture studies for cell attachment and proliferation. The HA released from the core region showed extended release up to 21 days for prolonged anti-adhesion effects. The attachment of adhesion-forming fibroblasts is reduced using the nanofibrous membrane from DNA assays and confocal microscopic observation of adhesion protein vinculin expression. The Ag released from the shell showed burst release to prevent E Coli and S. aureus infection immediately and prevent bacterial resistance to Ag. Minimum cytotoxicity was observed from Ag and IBU when fibroblasts were culture with the extraction medium of the nanofibrous membranes. The peritendinous anti-adhesion model in rabbits and the peritoneal anti-adhesion model in rats were used to test the efficacy of the anti-adhesion barriers as determined by gross observation, histology, and biomechanical tests. Within all membranes, the PEG/PCL/Ag/HA/IBU core-shell nanofibers showed the best reduction in cell attachment and proliferation when tested with fibroblasts in vitro. The PEG/PCL/Ag/HA/IBU nanofibrous membranes also showed significant improvement in preventing both peritendinous and peritoneal adhesions when compared with other groups and a commercial adhesion barrier film.

Keywords: anti-adhesion, electrospinning, hyaluronic acid, ibuprofen, nanofibers

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690 Dielectric and Impedance Spectroscopy of Samarium and Lanthanum Doped Barium Titanate at Room Temperature

Authors: Sukhleen Bindra Narang, Dalveer Kaur, Kunal Pubby

Abstract:

Dielectric ceramic samples in the BaO-Re2O3-TiO2 ternary system were synthesized with structural formula Ba2-xRe4+2x/3Ti8O24 where Re= rare earth metal and Re= Sm and La where x varies from 0.0 to 0.6 with step size 0.1. Polycrystalline samples were prepared by the conventional solid state reaction technique. The dielectric, electrical and impedance analysis of all the samples in the frequency range 1KHz- 1MHz at room temperature (25°C) have been done to get the understanding of electrical conduction and dielectric relaxation and their correlation. Dielectric response of the samples at lower frequencies shows dielectric dispersion while at higher frequencies it shows dielectric relaxation. The ac conductivity is well fitted by the Jonscher law (σac = σdc+Aωn). The spectroscopic data in the impedance plane confirms the existence of grain contribution to the relaxation. All the properties are found out to be function of frequency as well as the amount of substitution.

Keywords: dielectric ceramics, dielectric constant, loss tangent, AC conductivity, impedance spectroscopy

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689 SARS-CoV-2: Prediction of Critical Charged Amino Acid Mutations

Authors: Atlal El-Assaad

Abstract:

Viruses change with time through mutations and result in new variants that may persist or disappear. A Mutation refers to an actual change in the virus genetic sequence, and a variant is a viral genome that may contain one or more mutations. Critical mutations may cause the virus to be more transmissible, with high disease severity, and more vulnerable to diagnostics, therapeutics, and vaccines. Thus, variants carrying such mutations may increase the risk to human health and are considered variants of concern (VOC). Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) - the contagious in humans, positive-sense single-stranded RNA virus that caused coronavirus disease 2019 (COVID-19) - has been studied thoroughly, and several variants were revealed across the world with their corresponding mutations. SARS-CoV-2 has four structural proteins, known as the S (spike), E (envelope), M (membrane), and N (nucleocapsid) proteins, but prior study and vaccines development focused on genetic mutations in the S protein due to its vital role in allowing the virus to attach and fuse with the membrane of a host cell. Specifically, subunit S1 catalyzes attachment, whereas subunit S2 mediates fusion. In this perspective, we studied all charged amino acid mutations of the SARS-CoV-2 viral spike protein S1 when bound to Antibody CC12.1 in a crystal structure and assessed the effect of different mutations. We generated all missense mutants of SARS-CoV-2 protein amino acids (AAs) within the SARS-CoV-2:CC12.1 complex model. To generate the family of mutants in each complex, we mutated every charged amino acid with all other charged amino acids (Lysine (K), Arginine (R), Glutamic Acid (E), and Aspartic Acid (D)) and studied the new binding of the complex after each mutation. We applied Poisson-Boltzmann electrostatic calculations feeding into free energy calculations to determine the effect of each mutation on binding. After analyzing our data, we identified charged amino acids keys for binding. Furthermore, we validated those findings against published experimental genetic data. Our results are the first to propose in silico potential life-threatening mutations of SARS-CoV-2 beyond the present mutations found in the five common variants found worldwide.

Keywords: SARS-CoV-2, variant, ionic amino acid, protein-protein interactions, missense mutation, AESOP

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688 Novel p22-Monoclonal Antibody Based Blocking ELISA for the Detection of African Swine Fever Virus Antibodies in Serum

Authors: Ghebremedhin Tsegay, Weldu Tesfagaber, Yuanmao Zhu, Xijun He, Wan Wang, Zhenjiang Zhang, Encheng Sun, Jinya Zhang, Yuntao Guan, Fang Li, Renqiang Liu, Zhigao Bu, Dongming Zhao*

Abstract:

African swine fever (ASF) is a highly infectious viral disease of pigs, resulting in significant economic loss worldwide. As there is no approved vaccines and treatments, the control of ASF entirely depends on early diagnosis and culling of infected pigs. Thus, highly specific and sensitive diagnostic assays are required for accurate and early diagnosis of ASF virus (ASFV). Currently, only a few recombinant proteins have been tested and validated for use as reagents in ASF diagnostic assays. The most promising ones for ASFV antibody detection were p72, p30, p54, and pp62. So far, three ELISA kits based on these recombinant proteins have been commercialized. Due to the complex nature of the virus and variety forms of the disease, robust serodiagnostic assays are still required. ASFV p22 protein, encoded by KP177R gene, is located in the inner membrane of viral particle and appeared transiently in the plasma membrane early after virus infection. The p22 protein interacts with numerous cellular proteins, involved in processes of phagocytosis and endocytosis through different cellular pathways. However, p22 does not seem to be involved in virus replication or swine pathogenicity. In this study, E.coli expressed recombinant p22 protein was used to generate a monoclonal antibody (mAb), and its potential use for the development of blocking ELISA (bELISA) was evaluated. A total of 806 pig serum samples were tested to evaluate the bELISA. Acording the ROC (Reciever operating chracteristic) analysis, 100% sensitivity and 98.10% of specificity was recorded when the PI cut-off value was set at 47%. The novel assay was able to detect the antibodies as early as 9 days post infection. Finaly, a highly sensitive, specific and rapid novel p22-mAb based bELISA assay was developed, and optimized for detection of antibodies against genotype I and II ASFVs. It is a promising candidate for an early and acurate detection of the antibodies and is highly expected to have a valuable role in the containment and prevention of ASF.

Keywords: ASFV, blocking ELISA, diagnosis, monoclonal antibodies, sensitivity, specificity

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687 Electric Field Analysis of XLPE, Cross-Linked Polyethylene Covered Aerial Line and Insulator Lashing

Authors: Jyh-Cherng Gu, Ming-Ta Yang, Dai-Ling Tsai

Abstract:

Both sparse lashing and dense lashing are applied to secure overhead XLPE (cross-linked polyethylene) covered power lines on ceramic insulators or HDPE polymer insulators. The distribution of electric field in and among the lashing wires, the XLPE power lines and insulators in normal clean condition and when conducting materials such as salt, metal particles, dust, smoke or acidic smog are present is studied in this paper. The ANSYS Maxwell commercial software is used in this study for electric field analysis. Although the simulation analysis is performed assuming ideal conditions due to the constraints of the simulation software, the result may not be the same as in real situation but still be of sufficient practical values.

Keywords: electric field intensity, insulator, XLPE covered aerial line, empty

Procedia PDF Downloads 261
686 Investigation of the Effect of Phosphorous on the Flame Retardant Polyacrylonitrile Nanofiber

Authors: Mustafa Yılmaz, Ahmet Akar, Nesrin Köken, Nilgün Kızılcan

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Commercially available poly(acrylonitrile-co-vinyl acetate) P(AN-VA) or poly(acrylonitrile-co-methyl acrylate) P(AN-MA) are not satisfactory to meet the demand in flame and fire-resistance. In this work, vinylphosphonic acid is used during polymerization of acrylonitrile, vinyl acetate, methacrylic acid to produce fire-retardant polymers. These phosphorus containing polymers are successfully spun in the form of nanofibers. Properties such as water absorption of polymers are also determined and compared with commercial polymers.

Keywords: flame retardant, nanofiber, polyacrylonitrile, phosphorous compound, membrane

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685 A Novel Method to Manufacture Superhydrophobic and Insulating Polyester Nanofibers via a Meso-Porous Aerogel Powder

Authors: Z. Mazrouei-Sebdani, A. Khoddami, H. Hadadzadeh, M. Zarrebini

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Silica aerogels are well-known meso-porous materials with high specific surface area (500–1000 m2/g), high porosity (80–99.8%), and low density (0.003–0.8 g/cm3). However, the silica aerogels generally are highly brittle due to their nanoporous nature. Physical and mechanical properties of the silica aerogels can be enhanced by compounding with the fibers. Although some reports presented incorporation of the fibers into the sol, followed by further modification and drying stages, no information regarding the aerogel powders as filler in the polymeric fibers is available. In this research, waterglass based aerogel powder was prepared in the following steps: sol–gel process to prepare a gel, followed by subsequent washing with propan-2-ol, n-Hexane, and TMCS, then ambient pressure drying, and ball milling. Inspired by limited dust releasing, aerogel powder was introduced to the PET electrospinning solution in an attempt to create required bulk and surface structure for the nano fibers to improve their hydrophobic and insulation properties. The samples evaluation was carried out by measuring density, porosity, contact angle, sliding angle, heat transfer, FTIR, BET and SEM. According to the results, porous silica aerogel powder was fabricated with mean pore diameter of 24 nm and contact angle of 145.9º. The results indicated the usefulness of the aerogel powder confined into nano fibers to control surface roughness for manipulating superhydrophobic nanowebs with sliding angle of 5˚ and water contact angle of 147º. It can be due to a multi-scale surface roughness which was created by nanowebs structure itself and nano fibers surface irregularity in presence of the aerogels while a laye of fluorocarbon created low surface energy. The wettability of a solid substrate is an important property that is controlled by both the chemical composition and geometry of the surface. Also, a decreasing trend in the heat transfer was observed from 22% for the nano fibers without any aerogel powder to 8% for the nano fibers with 4% aerogel powder. The development of thermal insulating materials has become increasingly more important than ever in view of the fossil energy depletion and global warming that call for more demanding energy-saving practices.

Keywords: Superhydrophobicity, Insulation, Sol-gel, Surface energy, Roughness.

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684 Significance of Molecular Autophagic Pathway in Gaucher Disease Pathology

Authors: Ozlem Oral, Emre Taskin, Aysel Yuce, Serap Dokmeci, Devrim Gozuacik

Abstract:

Autophagy is an evolutionary conserved lysosome-dependent catabolic pathway, responsible for the degradation of long-lived proteins, abnormal aggregates and damaged organelles which cannot be degraded by the ubiquitin-proteasome system. Lysosomes degrade the substrates through the activity of lysosomal hydrolases and lysosomal membrane-bound proteins. Mutations in the coding region of these proteins cause malfunctional lysosomes, which contributes to the pathogenesis of lysosomal storage diseases. Gaucher disease is a lysosomal storage disease resulting from the mutation of a lysosomal membrane-associated glycoprotein called glucocerebrosidase and its cofactor saposin C. The disease leads to intracellular accumulation of glucosylceramide and other glycolipids. Because of the essential role of lysosomes in autophagic degradation, Gaucher disease may directly be linked to this pathway. In this study, we investigated the expression of autophagy and/or lysosome-related genes and proteins in fibroblast cells isolated from patients with different mutations. We carried out confocal microscopy analysis and examined autophagic flux by utilizing the differential pH sensitivities of RFP and GFP in mRFP-GFP-LC3 probe. We also evaluated lysosomal pH by active lysosome staining and lysosomal enzyme activity. Beside lysosomes, we also performed proteasomal activity and cell death analysis in patient samples. Our data showed significant attenuation in the expression of key autophagy-related genes and accumulation of their proteins in mutant cells. We found decreased the ability of autophagosomes to fuse with lysosomes, associated with elevated lysosomal pH and reduced lysosomal enzyme activity. Proteasomal degradation and cell death analysis showed reduced proteolytic activity of the proteasome, which consequently leads to increased susceptibility to cell death. Our data indicate that the major degradation pathways are affected by multifunctional lysosomes in mutant patient cells and may underlie in the mechanism of clinical severity of Gaucher patients. (This project is supported by TUBITAK-3501-National Young Researchers Career Development Program, Project No: 112T130).

Keywords: autophagy, Gaucher's disease, glucocerebrosidase, mutant fibroblasts

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683 Stabilizing of Lithium-Solid-Electrolyte Interfaces by Atomic Layer Deposition Prepared Nano-Interlayers for a Model All-Solid-State Battery

Authors: Rainer Goetz, Zahra Ahaliabadeh, Princess S. Llanos, Aliaksandr S. Bandarenka, Tanja Kallio

Abstract:

In order to understand the electrochemistry of all-solid-state batteries (ASSBs), the use of electrochemical equivalent circuits with a physical meaning is essential. A model battery is needed whose characterization is independent of the influence of the complex battery assembly. Lithium-Ion Conducting Glass-Ceramic (LICGC), a model solid electrolyte, is chosen for its stability in the air, but on the other hand, it is also well-known for its instability against metallic lithium upon direct contact. Hence, as a first step towards a model ASSB, the interface between lithium and the solid electrolyte (SE) is stabilized with thin (5 nm and 10 nm) coatings of titanium oxide (TO) and lithium titanium oxide (LTO). Impedance data shows that both materials are able to protect the SE surface from rapid degradation due to reducing lithium and, therefore, can serve as a protective interlayer on the anode side of a model ASSB.

Keywords: all-solid-state battery, lithium anode, solid electrolytes, interlayers

Procedia PDF Downloads 107
682 Fishing Waste: A Source of Valuable Products through Anaerobic Treatments

Authors: Luisa Maria Arrechea Fajardo, Luz Stella Cadavid Rodriguez

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Fish is one of the most commercialized foods worldwide. However, this industry only takes advantage of about 55% of the product's weight, the rest is converted into waste, which is mainly composed of viscera, gills, scales and spines. Consequently, if these wastes are not used or disposed of properly, they cause serious environmental impacts. This is the case of Tumaco (Colombia), the second largest producer of marine fisheries on the Colombian Pacific coast, where artisanal fishermen process more than 50% of the commercialized volume. There, fishing waste is disposed primarily in the ocean, causing negative impacts on the environment and society. Therefore, in the present research, a proposal was made to take advantage of fishing waste through anaerobic treatments, through which it is possible to obtain products with high added value from organic waste. The research was carried out in four stages. First, the production of volatile fatty acids (VFA) in semi-continuous 4L reactors was studied, evaluating three hydraulic retention times (HRT) (10, 7 and 5 days) with four organic loading rates (OLR) (16, 14, 12 and 10 gVS/L/day), the experiment was carried out for 150 days. Subsequently, biogas production was evaluated from the solid digestate generated in the VFA production reactors, initially evaluating the biochemical methane potential (BMP) of 4 total solid concentrations (1, 2, 4 and 6% TS), for 40 days and then, with the optimum TS concentration (2 gVS/L/day), 2 HRT (15 and 20 days) in semi-continuous reactors, were evaluated for 100 days. Finally, the integration of the processes was carried out with the best conditions found, a first phase of VFA production from fishing waste and a second phase of biogas production from unrecovered VFAs and unprocessed material Additionally, an VFA membrane extraction system was included. In the first phase, a liquid digestate with a concentration and VFA production yield of 59.04 gVFA/L and 0.527 gVFA/gVS, respectively, was obtained, with the best condition found (HRT:7 days and OLR: 16 gVS/L/día), where acetic acid and isobutyric acid were the predominant acids. In the second phase of biogas production, a BMP of 0.349 Nm3CH4/KgVS was reached, and it was found as best HRT 20 days. In the integration, the isovaleric, butyric and isobutyric acid were the VFA with the highest percentage of extraction, additionally a 106.67% increase in biogas production was achieved. This research shows that anaerobic treatments are a promising technology for an environmentally safe management of fishing waste and presents the basis of a possible biorefinery.

Keywords: biogas production, fishing waste, VFA membrane extraction, VFA production

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681 Micropower Composite Nanomaterials Based on Porous Silicon for Renewable Energy Sources

Authors: Alexey P. Antropov, Alexander V. Ragutkin, Nicolay A. Yashtulov

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The original controlled technology for power active nanocomposite membrane-electrode assembly engineering on the basis of porous silicon is presented. The functional nanocomposites were studied by electron microscopy and cyclic voltammetry methods. The application possibility of the obtained nanocomposites as high performance renewable energy sources for micro-power electronic devices is demonstrated.

Keywords: cyclic voltammetry, electron microscopy, nanotechnology, platinum-palladium nanocomposites, porous silicon, power activity, renewable energy sources

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680 Evaluation of the Efficacy of Titanium Alloy Dental Implants Coated by Bio-ceramic Apatite Wollastonite (Aw) and Hydroxyapatite (Ha) by Pulsed Laser Deposition

Authors: Betsy S. Thomas, Manjeet Marpara, K. M. Bhat

Abstract:

Introduction: After the initial enthusiasm and interest in hydroxyapatite products subsided due to dissolution of the coating and failure at the coating interface, this was a unique attempt to create a next generation of dental implant. Materials and Methods: The adhesion property of AW and HA coatings at various temperature by pulsed laser deposition was assessed on titanium plates. Moreover, AW/HA coated implants implanted in the femur of the rabbits was evaluated at various intervals. Results: Decohesion load was more for AW in scratch test and more bone formation around AW coated implants on histological evaluation. Discussion: AW coating by pulsed laser deposition was more adherent to the titanium surface and led to faster bone formation than HA. Conclusion: This experiment opined that AW coated by pulsed laser deposition seems to be a promising method in achieving bioactive coatings on titanium implants.

Keywords: surface coating, dental implants, osseo integration, biotechnology

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679 SiC Particulate-Reinforced SiC Composites Fabricated by PIP Method Using Highly Concentrated SiC Slurry

Authors: Jian Gu, Sea-Hoon Lee, Jun-Seop Kim

Abstract:

SiC particulate-reinforced SiC ceramic composites (SiCp/SiC) were successfully fabricated using polymer impregnation and pyrolysis (PIP) method. The effects of green density, infiltrated method, pyrolytic temperature, and heating rate on the densification behavior of the composites were investigated. SiCp/SiC particulate reinforced composites with high relative density up to 88.06% were fabricated after 4 PIP cycles using SiC pellets with high green density. The pellets were prepared by drying 62-70 vol.% aqueous SiC slurries, and the maximum relative density of the pellets was 75.5%. The hardness of the as-fabricated SiCp/SiCs was 21.05 GPa after 4 PIP cycles, which value increased to 23.99 GPa after a heat treatment at 2000℃. Excellent mechanical properties, thermal stability, and short processing time render the SiCp/SiC composite as a challenging candidate for the high-temperature application.

Keywords: high green density, mechanical property, polymer impregnation and pyrolysis, structural application

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678 An Overview of Sludge Utilization into Fired Clay Brick

Authors: Aeslina Binti Abdul Kadir, Ahmad Shayuti Bin Abdul Rahim

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Brick is one of the most common masonry units used as building material. Due to the demand, different types of waste have been investigated to be incorporated into the bricks. Many types of sludge have been incorporated in fired clay brick for example marble sludge, stone sludge, water sludge, sewage sludge, and ceramic sludge. The utilization of these waste materials in fired clay bricks usually has positive effects on the properties such as lightweight bricks with improved shrinkage, porosity, and strength. This paper reviews on utilization of different types of sludge wastes into fired clay bricks. Previous investigations have demonstrated positive effects on the physical and mechanical properties as well as less impact towards the environment. Thus, the utilizations of sludge waste could produce a good quality of brick and could be one of alternative disposal methods for the sludge wastes.

Keywords: fired clay brick, sludge waste, compressive strength, shrinkage, water absorption

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677 Waste Recovery: A Sustainable Way for Application of Solid Waste from WTP's in Building Materials

Authors: Flavio Araujo, Livia Dias, Fabiolla Lima, Paulo Scalize, Antonio Albuquerque

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Water treatment residues (WTR) are solid waste produced during drinking water treatment and have recently been seen as a reusable material. The aim of this research was show how to use the residue generated in a Water Treatment Plant, located in Goiania, Brazil, following the considerations of the law of solid waste to obtain normative parameters and consider sustainable alternatives for reincorporation of the residues in the productive chain for manufacturing various materials construction. In order to reduce the environmental liabilities generated by sanitation companies and discontinue unsustainable forms of disposal. The analyzes performed: Granulometry, Scanning Electron Microscopy and X-Ray Diffraction demonstrated the potential application of residues to replace the soil and sand, because it has characteristics compatible with small aggregate and can be used as feedstock for the manufacture of materials as ceramic and soil-cement bricks, mortars, interlocking floors and concrete artifacts.

Keywords: residue, sustainable, water treatment plants, WTR, WTP

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676 Hybrid Fermentation System for Improvement of Ergosterol Biosynthesis

Authors: Alexandra Tucaliuc, Alexandra C. Blaga, Anca I. Galaction, Lenuta Kloetzer, Dan Cascaval

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Ergosterol (ergosta-5,7,22-trien-3β-ol), also known as provitamin D2, is the precursor of vitamin D2 (ergocalciferol), because it is converted under UV radiation to this vitamin. The natural sources of ergosterol are mainly the yeasts (Saccharomyces sp., Candida sp.), but it can be also found in fungus (Claviceps sp.) or plants (orchids). In the yeasts cells, ergosterol is accumulated in membranes, especially in free form in the plasma membrane, but also as esters with fatty acids in membrane lipids. The chemical synthesis of ergosterol does not represent an efficient method for its production, in these circumstances, the most attractive alternative for producing ergosterol at larger-scale remains the aerobic fermentation using S. cerevisiae on glucose or by-products from agriculture of food industry as substrates, in batch or fed-batch operating systems. The aim of this work is to analyze comparatively the influence of aeration efficiency on ergosterol production by S. cerevisiae in batch and fed-batch fermentations, by considering different levels of mixing intensity, aeration rate, and n-dodecane concentration. The effects of the studied factors are quantitatively described by means of the mathematical correlations proposed for each of the two fermentation systems, valid both for the absence and presence of oxygen-vector inside the broth. The experiments were carried out in a laboratory stirred bioreactor, provided with computer-controlled and recorded parameters. n-Dodecane was used as oxygen-vector and the ergosterol content inside the yeasts cells has been considered at the fermentation moment related to the maximum concentration of ergosterol, 9 hrs for batch process and 20 hrs for fed-batch one. Ergosterol biosynthesis is strongly dependent on the dissolved oxygen concentration. The hydrocarbon concentration exhibits a significant influence on ergosterol production mainly by accelerating the oxygen transfer rate. Regardless of n-dodecane addition, by maintaining the glucose concentration at a constant level in the fed-batch process, the amount of ergosterol accumulated into the yeasts cells has been almost tripled. In the presence of hydrocarbon, the ergosterol concentration increased by over 50%. The value of oxygen-vector concentration corresponding to the maximum level of ergosterol depends mainly on biomass concentration, due to its negative influences on broth viscosity and interfacial phenomena of air bubbles blockage through the adsorption of hydrocarbon droplets–yeast cells associations. Therefore, for the batch process, the maximum ergosterol amount was reached for 5% vol. n-dodecane, while for the fed-batch process for 10% vol. hydrocarbon.

Keywords: bioreactors, ergosterol, fermentation, oxygen-vector

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675 Production and Characterization of Implant Material Produced by Using Electroless Ni Plated Al2O3-Co-Cr-Ti Powders

Authors: Ahmet Yonetken, Ayhan Erol

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The microstructure, mechanical properties and corrosion characteristics of Ni plated %10Al2O3-%40Co-%20Cr and %10Ti powders were investigated using specimens produced by tube furnace sintering at 800-1200°C temperature. A uniform nickel layer on Al2O3-Co-Cr and Ti powders was deposited prior to sintering using electroless plating technique. A composite consisting of quintet additions, a metallic phase, Ti,Cr and Co including a ceramic phase, alumina, within a matrix of Ni has been prepared under Ar shroud and then tube furnace sintered. XRD, SEM (Scanning Electron Microscope), corrosion behavior in acidic media were investigated to characterize the properties of the specimens. Experimental results carried out for composition (%10Al2O3-%40Co-%20Cr- %10Ti)20Ni at 1200°C suggest that the best properties as 312.18HV were obtained at 1200°C.

Keywords: sintering, intermetallic, Electroless nickel plating, composite

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674 ICAM-2, A Protein of Antitumor Immune Response in Mekong Giant Catfish (Pangasianodon gigas)

Authors: Jiraporn Rojtinnakorn

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ICAM-2 (intercellular adhesion molecule 2) or CD102 (Cluster of Differentiation 102) is type I trans-membrane glycoproteins, composing 2-9 immunoglobulin-like C2-type domains. ICAM-2 plays the particular role in immune response and cell surveillance. It is concerned in innate and specific immunity, cell survival signal, apoptosis, and anticancer. EST clone of ICAM-2, from P. gigas blood cell EST libraries, showed high identity to human ICAM-2 (92%) with conserve region of ICAM N-terminal domain and part of Ig superfamily. Gene and protein of ICAM-2 has been founded in mammals. This is the first report of ICAM-2 in fish.

Keywords: ICAM-2, CD102, Pangasianodon gigas, antitumor

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673 Moderate Electric Field Influence on Carotenoids Extraction Time from Heterochlorella luteoviridis

Authors: Débora P. Jaeschke, Eduardo A. Merlo, Rosane Rech, Giovana D. Mercali, Ligia D. F. Marczak

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Carotenoids are high value added pigments that can be alternatively extracted from some microalgae species. However, the application of carotenoids synthetized by microalgae is still limited due to the utilization of organic toxic solvents. In this context, studies involving alternative extraction methods have been conducted with more sustainable solvents to replace and reduce the solvent volume and the extraction time. The aim of the present work was to evaluate the extraction time of carotenoids from the microalgae Heterochlorella luteoviridis using moderate electric field (MEF) as a pre-treatment to the extraction. The extraction methodology consisted of a pre-treatment in the presence of MEF (180 V) and ethanol (25 %, v/v) for 10 min, followed by a diffusive step performed for 50 min using a higher ethanol concentration (75 %, v/v). The extraction experiments were conducted at 30 °C and, to keep the temperature at this value, it was used an extraction cell with a water jacket that was connected to a water bath. Also, to enable the evaluation of MEF effect on the extraction, control experiments were performed using the same cell and conditions without voltage application. During the extraction experiments, samples were withdrawn at 1, 5 and 10 min of the pre-treatment and at 1, 5, 30, 40 and 50 min of the diffusive step. Samples were, then, centrifuged and carotenoids analyses were performed in the supernatant. Furthermore, an exhaustive extraction with ethyl acetate and methanol was performed, and the carotenoids content found for this analyses was considered as the total carotenoids content of the microalgae. The results showed that the application of MEF as a pre-treatment to the extraction influenced the extraction yield and the extraction time during the diffusive step; after the MEF pre-treatment and 50 min of the diffusive step, it was possible to extract up to 60 % of the total carotenoids content. Also, results found for carotenoids concentration of the extracts withdrawn at 5 and 30 min of the diffusive step did not presented statistical difference, meaning that carotenoids diffusion occurs mainly in the very beginning of the extraction. On the other hand, the results for control experiments showed that carotenoids diffusion occurs mostly during 30 min of the diffusive step, which evidenced MEF effect on the extraction time. Moreover, carotenoids concentration on samples withdrawn during the pre-treatment (1, 5 and 10 min) were below the quantification limit of the analyses, indicating that the extraction occurred in the diffusive step, when ethanol (75 %, v/v) was added to the medium. It is possible that MEF promoted cell membrane permeabilization and, when ethanol (75 %) was added, carotenoids interacted with the solvent and the diffusion occurred easily. Based on the results, it is possible to infer that MEF promoted the decrease of carotenoids extraction time due to the increasing of the permeability of the cell membrane which facilitates the diffusion from the cell to the medium.

Keywords: moderate electric field (MEF), pigments, microalgae, ethanol

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672 Cytokine Profiling in Cultured Endometrial Cells after Hormonal Treatment

Authors: Mark Gavriel, Ariel J. Jaffa, Dan Grisaru, David Elad

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The human endometrium-myometrium interface (EMI) is the uterine inner barrier without a separatig layer. It is composed of endometrial epithelial cells (EEC) and endometrial stromal cells (ESC) in the endometrium and myometrial smooth muscle cells (MSMC) in the myometrium. The EMI undergoes structural remodeling during the menstruation cycle which are essential for human reproduction. Recently, we co-cultured a layer-by-layer in vitro model of EEC, ESC and MSMC on a synthetic membrane for mechanobiology experiments. We also treated the model with progesterone and β-estradiol in order to mimic the in vivo receptive uterus In the present study we analyzed the cytokines profile in a single layer of EEC the hormonal treated in vitro model of the EMI. The methodologies of this research include simple tissue-engineering . First, we cultured commercial EEC (RL95-2, ATCC® CRL-1671™) in 24-wellplate. Then, we applied an hormonal stimuli protocol with 17-β-estradiol and progesterone in time dependent concentration according to the human physiology that mimics the menstrual cycle. We collected cell supernatant samples of control, pre-ovulation, ovulation and post-ovulaton periods for analysis of the secreted proteins and cytokines. The cytokine profiling was performed using the Proteome Profiler Human XL Cytokine Array Kit (R&D Systems, Inc., USA) that can detect105 human soluble cytokines. The relative quantification of all the cytokines will be analyzed using xMAP – LUMINEX. We conducted a fishing expedition with the 4 membranes Proteome Profiler. We processed the images, quantified the spots intensity and normalized these values by the negative control and reference spots at the membrane. Analyses of the relative quantities that reflected change higher than 5% of the control points of the kit revealed the The results clearly showed that there are significant changes in the cytokine level for inflammation and angiogenesis pathways. Analysis of tissue-engineered models of the uterine wall will enable deeper investigation of molecular and biomechanical aspects of early reproductive stages (e.g. the window of implantation) or developments of pathologies.

Keywords: tissue-engineering, hormonal stimuli, reproduction, multi-layer uterine model, progesterone, β-estradiol, receptive uterine model, fertility

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671 Learning from Dendrites: Improving the Point Neuron Model

Authors: Alexander Vandesompele, Joni Dambre

Abstract:

The diversity in dendritic arborization, as first illustrated by Santiago Ramon y Cajal, has always suggested a role for dendrites in the functionality of neurons. In the past decades, thanks to new recording techniques and optical stimulation methods, it has become clear that dendrites are not merely passive electrical components. They are observed to integrate inputs in a non-linear fashion and actively participate in computations. Regardless, in simulations of neural networks dendritic structure and functionality are often overlooked. Especially in a machine learning context, when designing artificial neural networks, point neuron models such as the leaky-integrate-and-fire (LIF) model are dominant. These models mimic the integration of inputs at the neuron soma, and ignore the existence of dendrites. In this work, the LIF point neuron model is extended with a simple form of dendritic computation. This gives the LIF neuron increased capacity to discriminate spatiotemporal input sequences, a dendritic functionality as observed in another study. Simulations of the spiking neurons are performed using the Bindsnet framework. In the common LIF model, incoming synapses are independent. Here, we introduce a dependency between incoming synapses such that the post-synaptic impact of a spike is not only determined by the weight of the synapse, but also by the activity of other synapses. This is a form of short term plasticity where synapses are potentiated or depressed by the preceding activity of neighbouring synapses. This is a straightforward way to prevent inputs from simply summing linearly at the soma. To implement this, each pair of synapses on a neuron is assigned a variable,representing the synaptic relation. This variable determines the magnitude ofthe short term plasticity. These variables can be chosen randomly or, more interestingly, can be learned using a form of Hebbian learning. We use Spike-Time-Dependent-Plasticity (STDP), commonly used to learn synaptic strength magnitudes. If all neurons in a layer receive the same input, they tend to learn the same through STDP. Adding inhibitory connections between the neurons creates a winner-take-all (WTA) network. This causes the different neurons to learn different input sequences. To illustrate the impact of the proposed dendritic mechanism, even without learning, we attach five input neurons to two output neurons. One output neuron isa regular LIF neuron, the other output neuron is a LIF neuron with dendritic relationships. Then, the five input neurons are allowed to fire in a particular order. The membrane potentials are reset and subsequently the five input neurons are fired in the reversed order. As the regular LIF neuron linearly integrates its inputs at the soma, the membrane potential response to both sequences is similar in magnitude. In the other output neuron, due to the dendritic mechanism, the membrane potential response is different for both sequences. Hence, the dendritic mechanism improves the neuron’s capacity for discriminating spa-tiotemporal sequences. Dendritic computations improve LIF neurons even if the relationships between synapses are established randomly. Ideally however, a learning rule is used to improve the dendritic relationships based on input data. It is possible to learn synaptic strength with STDP, to make a neuron more sensitive to its input. Similarly, it is possible to learn dendritic relationships with STDP, to make the neuron more sensitive to spatiotemporal input sequences. Feeding structured data to a WTA network with dendritic computation leads to a significantly higher number of discriminated input patterns. Without the dendritic computation, output neurons are less specific and may, for instance, be activated by a sequence in reverse order.

Keywords: dendritic computation, spiking neural networks, point neuron model

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670 The Quantitative Optical Modulation of Dopamine Receptor-Mediated Endocytosis Using an Optogenetic System

Authors: Qiaoyue Kuang, Yang Li, Mizuki Endo, Takeaki Ozawa

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G protein-coupled receptors (GPCR) are the largest family of receptor proteins that detect molecules outside the cell and activate cellular responses. Of the GPCRs, dopamine receptors, which recognize extracellular dopamine, are essential to mammals due to their roles in numerous physiological events, including autonomic movement, hormonal regulation, emotions, and the reward system in the brain. To precisely understand the physiological roles of dopamine receptors, it is important to spatiotemporally control the signaling mediated by dopamine receptors, which is strongly dependent on their surface expression. Conventionally, chemical-induced interactions were applied to trigger the endocytosis of cell surface receptors. However, these methods were subjected to diffusion and therefore lacked temporal and special precision. To further understand the receptor-mediated signaling and to control the plasma membrane expression of receptors, an optogenetic tool called E-fragment was developed. The C-terminus of a light-sensitive photosensory protein cyptochrome2 (CRY2) was attached to β-Arrestin, and the E-fragment was generated by fusing the C-terminal peptide of vasopressin receptor (V2R) to CRY2’s binding partner protein CIB. The CRY2-CIB heterodimerization triggered by blue light stimulation brings β-Arrestin to the vicinity of membrane receptors and results in receptor endocytosis. In this study, the E-fragment system was applied to dopamine receptors 1 and 2 (DRD1 and DRD2) to control dopamine signaling. First, confocal fluorescence microscope observation qualitatively confirmed the light-induced endocytosis of E-fragment fused receptors. Second, NanoBiT bioluminescence assay verified quantitatively that the surface amount of E-fragment labeled receptors decreased after light treatment. Finally, GloSensor bioluminescence assay results suggested that the E-fragment-dependent receptor light-induced endocytosis decreased cAMP production in DRD1 signaling and attenuated the inhibition effect of DRD2 on cAMP production. The developed optogenetic tool was able to induce receptor endocytosis by external light, providing opportunities to further understand numerous physiological activities by controlling receptor-mediated signaling spatiotemporally.

Keywords: dopamine receptors, endocytosis, G protein-coupled receptors, optogenetics

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669 Embryotoxicity of Nano-Iron Oxide (Fe2O3) to Bio-Indicator of Pollution of Land Helix Aspersa

Authors: S. Besnaci, S. Bensoltane, H. Locif, S. Saadi

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To validate an ecotoxicological approach to assessing toxicological effects caused by the oxide powder of nano-iron Fe2O3, we searched in the ecotoxicology laboratory cell bodies bio accumulators and bio-indicators of soil pollution the snail Helix aspersa. In this study, we evaluated the toxicity of nano Fe2O3 during a very sensitive phase of development H.aspersa (embryonic stage). During embryonic development, we observed in treated with various concentrations of nano Fe2O3 (1.25 g/l, 1.5 g/l, and 2 g/l) compared to control, the deformation of the membrane of the egg and accumulation of this molecule at the rear of the egg proven by the photographs, as with the influence on the hatching percentage.

Keywords: eggs, embryotoxicity, Fe2O3, Helix aspersa, nanoparticles

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668 Tensile and Flexural Behavior of Particulate Filled/Polymer Matrix Composites

Authors: M. Alsaadi, A. Erkliğ, M. Bulut

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This paper experimentally investigates the flexural and tensile properties of the industrial wastes sewage sludge ash (SSA) and fly ash (FA), and conventional ceramic powder silicon carbide (SiC) filled polyester composites. Four weight fractions (5, 10, 15 and 20 wt%) for each micro filler were used for production of composites. Then, test samples were produced according to ASTM. The resulting degree of particle dispersion in the polymer matrix was visualized by using scanning electron microscope (SEM). Results from this study showed that the tensile strength increased up to its maximum value at filler content 5 wt% of SSA, FA and SiC. Flexural strength increased with addition of particulate filler up to its maximum value at filler content 5 wt% of SSA and FA while for SiC decreased for all weight fractions gradually. The addition of SSA, FA and SiC fillers resulted in increase of tensile and flexural modulus for all the particulate composites. Industrial waste SSA can be used as an additive with polymer to produce composite materials.

Keywords: particle-reinforcement, sewage sludge ash, polymer matrix composites, mechanical properties

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667 Sacidava and Its Role of Military Outpost in the Moesian Sector of the Danube Limes: Animal Food Resources and Landscape Reconstruction

Authors: Margareta Simina Stanc, Aurel Mototolea, Tiberiu Potarniche

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Sacidava archeological site is located in Dobrudja region, Romania, on a hill on the right bank of the Danube - the Musait point, located at about 5 km north-east from Dunareni village. The place-name documents the fact that, prior to the Roman conquest, in the area, there was a Getic settlement. The location of the Sacidava was made possible by corroborating the data provided by the ancient sources with the epigraphic documents (the milial pillar during the time of Emperor Decius). The tegular findings attest that an infantry unit, cohors I Cilicum milliaria equitata, as well as detachments from Legio V Macedonica and Legio XI Claudia, were confined to Sacidava. During the period of the Dominion, the garrison of the fortification is the host of a cavalry unit: cuneus equitum scutariorum. In the immediate vicinity to the Roman fortress, to the east, were identified two other fortifications: a Getic settlement (4th-1st century B.C.) and an Early Medieval settlement (9th-10th century A.C.). The archaeological material recovered during the research is represented by ceramic forms such as amphoras, jugs, pots, cups, plates, to which are added oil lamps, some of them typologically new at the time of discovery. Local ceramic shapes were also founded, worked by hand or by wheel, considered un-Romanized or in the course of Romanization. During the time of the Principality, Sacidava it represented an important military outpost serving mainly the city of Tropaeum Traiani, controlling also the supply and transport on the Danube limes in the Moesic sector. This role will determine the development of the fortress and the appearance of extramuros civil structures, thus becoming an important landmark during the 5th-6th centuries A.C., becoming a representation of power of the Roman empire in an area of continuous conflict. During recent archaeological researches, faunal remains were recovered, and their analysis allowed to estimate the animal resources and subsistence practices (animal husbandry, hunting, fishing) in the settlement. The methodology was specific to archaeozoology, mainly consisting of anatomical, taxonomical, and taphonomical identifications, recording, and quantification of the data. The remains of domestic mammals have the highest proportion indicating the importance of animal husbandry; the predominant species are Bos taurus, Ovis aries/Capra hircus, and Sus domesticus. Fishing and hunting were of secondary importance in the subsistence economy of the community. Wild boar and the red deer were the most frequently hunted species. Just a few fish bones were recovered. Thus, the ancient city of Sacidava is proving to be an important element of cultural heritage of the south-eastern part of Romania, for whose conservation and enhancement efforts must be made, especially by landscape reconstruction.

Keywords: archaeozoology, landscape reconstruction, limes, military outpost

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666 Influence of Substitution on Structure of Tin Lantanium Pyrochlore La₂₋ₓSrₓSn₂O₇₋δ(0 ≤ x ≤ 0.25) Solid-Oxide Fuel Cells

Authors: Bounar Nedjemeddine

Abstract:

Materials with the pyrochlore lattice structure have attracted much recent attention due to their wide applications in ceramic thermal barrier coatings, high-permittivity dielectrics, and potential solid electrolytes in solid-oxide fuel cells. The work described in this paper is devoted to the synthesis and characterization of a pyrochlore structure based on lanthanum (La₂O₃) and tin (SnO₂) oxides of general formula La₂Sn₂O₇, substituted by Sr at the site La. Their structures were determined from X-ray powder diffraction using CELFER analysis. All the compositions present the space group Fd-3m. The substitution of La by Sr in the La₂Sn₂O₇ compound causes a variation of the cell parameters. The difference in charge between La³⁺ and Sr²⁺ and the difference in size cause the cell parameters to decrease from a=10.7165 A° to a=10.6848 A° for the substitution rates (x = 0.05, 0.1, 0.15 ...), which leads to a decrease in the volume of the mesh. For a substitution rate x = 0.25, there is an increase in the cell parameters (a=10.7035A°), which can be explained by a competitiveness of the size effect and the presence of a gap in the structure which go in the opposite direction.

Keywords: solid-oxide fuel cells, structure, pyrochlore, X-ray diffraction

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665 Prospects of Regenerative Medicine with Human Allogeneic Adipose Tissue-Derived Mesenchymal Stem Cell Sheets: Achievements and Future Outlook in Clinical Trials for Myopic Chorioretinal Atrophy

Authors: Norimichi Nagano, Yoshio Hirano, Tsutomu Yasukawa

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Mesenchymal stem cells are thought to confer neuroprotection, facilitate tissue regeneration and exert their effects on retinal degenerative diseases, however, adverse events such as proliferative vitreoretinopathy and preretinal membrane disease associated with cell suspension transplantation have also been reported. We have recently developed human (allogeneic) adipose tissue-derived mesenchymal stem cell (adMSC) sheets through our proprietary sheet transformation technique, which could potentially mitigate these adverse events. To clarify the properties of our adMSC sheets named PAL-222, we performed in vitro studies such as viability testing, cytokine secretions by ELISA, immunohistochemical study, and migration assay. The viability of the cells exceeded 70%. Vascular Endothelial Growth Factor (VEGF) and Pigment Epithelium-Derived Factor (PEDF), which are quite important cytokines for the retinal area, were observed. PAL-222 expressed type I collagen, a strength marker, type IV collagen, a marker of the basement membrane, and elastin, an elasticity marker. Finally, the migration assay was performed and showed negative, which means that PAL-222 is stably kept in the topical area and does not come to pieces. Next, to evaluate the efficacy in vivo, we transplanted PAL-222 into the subretinal space of the eye of Royal College of Surgeons rats with congenital retinal degeneration and assessed it for three weeks after transplantation. We confirmed that PAL-222 suppressed the decrease in the thickness of the outer nuclear layer, which means that the photoreceptor protective effect treated with PAL-222 was significantly higher than that in the sham group. (p < 0.01). This finding demonstrates that PAL-222 showed their retinoprotective effect in a model of congenital retinal degeneration. As the study suggested the efficacy of PAL-222 in both in vitro and in vivo studies, we are presently engaged in clinical trials of PAL-222 for myopic chorioretinal atrophy, which is one of the retinal degenerative diseases, for the purpose of regenerative medicine.

Keywords: cell sheet, clinical trial, mesenchymal stem cell, myopic chorioretinal atrophy

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664 Study of Durability of Porous Polymer Materials, Glass-Fiber-Reinforced Polyurethane Foam (R-PUF) in MarkIII Containment Membrane System

Authors: Florent Cerdan, Anne-Gaëlle Denay, Annette Roy, Jean-Claude Grandidier, Éric Laine

Abstract:

The insulation of MarkIII membrane of the Liquid Natural Gas Carriers (LNGC) consists of a load- bearing system made of panels in reinforced polyurethane foam (R-PUF). During the shipping, the cargo containment shall be potentially subject to risk events which can be water leakage through the wall ballast tank. The aim of these present works is to further develop understanding of water transfer mechanisms and water effect on properties of R-PUF. This multi-scale approach contributes to improve the durability. Macroscale / Mesoscale Firstly, the use of the gravimetric technique has allowed to define, at room temperature, the water transfer mechanisms and kinetic diffusion, in the R-PUF. The solubility follows a first kinetic fast growing connected to the water absorption by the micro-porosity, and then evolves linearly slowly, this second stage is connected to molecular diffusion and dissolution of water in the dense membranes polyurethane. Secondly, in the purpose of improving the understanding of the transfer mechanism, the study of the evolution of the buoyant force has been established. It allowed to identify the effect of the balance of total and partial pressure of mixture gas contained in pores surface. Mesoscale / Microscale The differential scanning calorimetry (DSC) and Dynamical Mechanical Analysis (DMA), have been used to investigate the hydration of the hard and soft segments of the polyurethane matrix. The purpose was to identify the sensitivity of these two phases. It been shown that the glass transition temperatures shifts towards the low temperatures when the solubility of the water increases. These observations permit to conclude to a plasticization of the polymer matrix. Microscale The Fourier Transform Infrared (FTIR) study has been used to investigate the characterization of functional groups on the edge, the center and mid-way of the sample according the duration of submersion. More water there is in the material, more the water fix themselves on the urethanes groups and more specifically on amide groups. The pic of C=O urethane shifts at lower frequencies quickly before 24 hours of submersion then grows slowly. The intensity of the pic decreases more flatly after that.

Keywords: porous materials, water sorption, glass transition temperature, DSC, DMA, FTIR, transfer mechanisms

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663 Dielectric Properties of Ternary Composite Based on Complex Perovskite Oxides Synthesized by Semi-Wet Route

Authors: K. D. Mandal, Anil Kumar Mourya, Ankur Khare

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Ceramics of 0.6 CaCu3TiO12 – 0.2 Bi2/3Cu3TiO12 – 0.2 Y2/3Cu3TiO12 (i.e. 0.6CCTO-0.2BCTO-0.2 YCTO) were prepared via semi - wet route. The phase structure of the sample was identified by X-Ray diffraction. The micro structure of the sample was observed by SEM, which displays grains of different shapes having diameter in range of 2 µm–4 µm. We have studied the frequency and temperature dependence of permittivity and impedance of the compound with LCR Meter in the range of 100 Hz–1 MHz and 300–500 K. The material shows its highest dielectric constant (428000) at 100 Hz and 368 K. The material shows Debye–like relaxation and their dielectric constant are independent of frequency and temperature over a wide range. The sample shows two electrical responses in impedance formalism, indicating that there are two distinct contributions. We attribute them to grain and grain boundaries in the ceramic sample and explain the dielectric behaviors by Maxwell–Wagner relaxation arising at the interfaces between grain and their boundaries.

Keywords: complex perovskite, ceramics composite, impedance study, SEM

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662 Microstructure and Mechanical Evaluation of PMMA/Al₂O₃ Nanocomposite Fabricated via Friction Stir Processing

Authors: Reham K. El Sawah, N. S. M. El-Tayeb

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This study aims to produce a polymer matrix composite reinforced with Al₂O₃ nanoparticles in order to enhance the mechanical properties of PMMA. The composite was fabricated via Friction stir processing to ensure homogenous dispersion of Al₂O₃ nanoparticles in the polymer, and the processing was submerged to prevent the sputtering of nanoparticles. The surface quality, microstructure, impact energy and hardness of the prepared samples were investigated. Good surface quality and dispersion of nanoparticles were attained through employing sufficient processing conditions. The experimental results indicated that as the percentage of nanoparticles increased, the impact energy and hardness increased, reaching 2 kJ/m2 and 14.7 HV at a nanoparticle concentration of 25%, which means that the toughness and the hardness of the polymer-ceramic produced composite is higher than unprocessed PMMA by 66% and 33% respectively.

Keywords: friction stir processing, polymer matrix nanocomposite, mechanical properties, microstructure

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