Search results for: collagen membrane

Authors: Shende Suraj Balu, A. B. Deoghare, K. M. Pandey

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The human middle ear (ME) is a delicate and vital organ. It has a complex structure that performs various functions such as receiving sound pressure and producing vibrations of eardrum and propagating it to inner ear. It consists of Tympanic Membrane (TM), three auditory ossicles, various ligament structures and muscles. Incidents such as traumata, infections, ossification of ossicular structures and other pathologies may damage the ME organs. The conditions can be surgically treated by employing prosthesis. However, the suitability of the prosthesis needs to be examined in advance prior to the surgery. Few decades ago, this issue was addressed and analyzed by developing an equivalent representation either in the form of spring mass system, electrical system using R-L-C circuit or developing an approximated CAD model. But, nowadays a three-dimensional ME model can be constructed using micro X-Ray Computed Tomography (μCT) scan data. Moreover, the concern about patient specific integrity pertaining to the disease can be examined well in advance. The current research work emphasizes to develop the ME model from the stacks of μCT images which are used as input file to MIMICS Research 19.0 (Materialise Interactive Medical Image Control System) software. A stack of CT images is converted into geometrical surface model to build accurate morphology of ME. The work is further extended to understand the dynamic behaviour of Harmonic response of the stapes footplate and umbo for different sound pressure levels applied at lateral side of eardrum using finite element approach. The pathological condition Cholesteatoma of ME is investigated to obtain peak to peak displacement of stapes footplate and umbo. Apart from this condition, other pathologies, mainly, changes in the stiffness of stapedial ligament, TM thickness and ossicular chain separation and fixation are also explored. The developed model of ME for pathologies is validated by comparing the results available in the literatures and also with the results of a normal ME to calculate the percentage loss in hearing capability.

Keywords: computed tomography (μCT), human middle ear (ME), harmonic response, pathologies, tympanic membrane (TM)

Procedia PDF Downloads 175

Authors: S. N. Moghaddas Tafreshi, S. M. Amin Ghotbi

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This paper presents a numerical study for investigating the effectiveness of geocell reinforcement in reducing pressure and settlement over EPS geofoam blocks in road embankments. A 3-D FEM model of soil and geofoam was created in ABAQUS, and geocell was also modeled realistically using membrane elements. The accuracy of the model was tested by comparing its results with previous works. Sensitivity analyses showed that reinforcing the soil cover with geocell has a significant influence on the reduction of imposed stresses over geofoam and consequently decreasing its deformation.

Keywords: EPS geofoam, geocell, reinforcement, road embankments, lightweight fill

Procedia PDF Downloads 273

Authors: L. Hamra, J. F. Demonceau, V. Denoël

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The aim of this paper is to study a beam extracted from a frame and subjected to blast loading. The demand of ductility depends on six dimensionless parameters: two related to the blast loading, two referring to the bending behavior of the beam and two corresponding to the dynamic behavior of the rest of the structure. We develop a full analytical procedure that provides the ductility demand as a function of these six dimensionless parameters.

Keywords: analytical procedure, blast loading, membrane force, P-I diagram

Procedia PDF Downloads 427

Authors: S. A. Abdellatef, A. Taniguchi, Namiki, Tsukuba, Ibaraki

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The alterations in the physicochemical characteristics of bio-materials are renowned for their impact in cellular behaviors. Surface chemistry and substratum topography are separately considered as mutable characteristics with deep impact on the overall cell behaviors. In our recent work, we examined the manipulation of the physical cues on hepatic cellular behaviors. We have proven that the geometrical or dimensional characteristics of nano features are essential for the optimum hepatocellular functions. While here, the collective impact of both physical and chemical cues on hepatocellular behaviors was investigated. On which RGD peptide was immobilized on a TiO2 nano pattern that imitates the hierarchically extend collagen nano fibrillar structures. The hepatocytes morphological and functional changes induced by simultaneously combining the diversified cues were investigated. TiO2 substrates that integrate nano topography with the adhesive peptide motif (RGD) had showed an increase in the hepatocellular functionality to the maximum extent. While a significant enhancement in expression of these liver specific markers on RGD coated surfaces were observed compared to uncoated substrates regardless of topography. Consequently in depth understanding of the relationship between various kind of cues and hepatocytes behaviors would be a paving step in the application of tissue engineering and bio reactor technology.

Keywords: biomaterial, tiO2, hepG2, RGD

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Authors: G. G. Kagramanov, E. N. Farnosova, Linn Maung Maung

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The technologies of ion exchange and nanofiltration can be used for treatment of wastewater containing copper and other heavy metal ions to decrease the environmental risks. Nanofiltration characteristics under water treatment of heavy metals have been studied. The influence of main technical process parameters - pressure, temperature, concentration and pH value of the initial solution on flux and rejection of nanofiltration membranes has been considered. And ion exchange capacities of resins in removal of heavy metal ions from wastewater have been determined.

Keywords: exchange capacity, heavy metals, ion exchange, membrane separation, nanofiltration

Procedia PDF Downloads 286

Authors: Nor Khaizan Anuar, Nur Karimah Aziz, Tin Wui Wong, Ahmad Sazali Hamzah, Wan Rozita Wan Engah

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The physicochemical attributes and wound healing activity of pectin hydrogel in rat models, following partial thickness thermal injury were investigated. The pectin hydrogel was prepared by solvent evaporation method with the aid of glutaraldehyde as crosslinking agent and glycerol as plasticizer. The physicochemical properties were mainly evaluated using differential scanning calorimetry (DSC) and Fourier transform infrared (FTIR) spectroscopy, while the wound healing activity was examined by the macroscopic images, wound size reduction and histological evaluation using haematoxylin and eosin (H&E) stain for 14 days. The DSC and FTIR analysis suggested that pectin hydrogel exhibited higher extent of polymer-polymer interaction at O-H functional group in comparison to the unprocessed pectin. This was indicated by the increase of endothermic enthalpy values from 139.35 ± 13.06 J/g of unprocessed pectin to 156.23 ± 2.86 J/g of pectin hydrogel, as well as the decrease of FTIR wavenumber corresponding to O-H at 3432.07 ± 0.49 cm-1 of unprocessed pectin to 3412.62 ± 13.06 cm-1 of pectin hydrogel. Rats treated with pectin hydrogel had significantly smaller wound size (Student’s t-test, p<0.05) when compared to the untreated group starting from day 7 until day 14. H&E staining indicated that wounds received pectin hydrogel had more fibroblasts, blood vessels and collagen bundles on day 14 in comparison to the untreated rats.

Keywords: pectin, physicochemical, rats, wound

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Authors: Asim Laeeq Khan, Mazhar Amjad Gilani, Tayub Raza

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The use of fossil fuels for energy generation leads to the emission of greenhouse gases particularly CO2 into the atmosphere. To date, several techniques have been proposed for the efficient removal of CO2 from flue gas mixtures. Membrane technology is a promising choice due to its several inherent advantages such as low capital cost, high energy efficiency, and low ecological footprint. One of the goals in the development of membranes is to achieve high permeability and selectivity. Mixed matrix membranes comprising of inorganic fillers embedded in polymer matrix are a class of membranes that have showed improved separation properties. One of the biggest challenges in the commercialization if mixed matrix membranes are the removal of non-selective voids existing at the polymer-filler interface. In this work, mixed matrix membranes were prepared using polysulfone as polymer matrix and ordered mesoporous MCM-41 as filler materials. A new approach to removing the interfacial voids was developed by introducing room temperature ionic (RTIL) at the polymer-filler interface. The results showed that the imidazolium based RTIL not only provided wettability characteristics but also helped in further improving the separation properties. The removal of interfacial voids and good contact between polymer and filler was verified by SEM measurement. The synthesized membranes were tested in a custom built gas permeation set-up for the measurement of gas permeability and ideal gas selectivity. The results showed that the mixed matrix membranes showed significantly higher CO2 permeability in comparison to the pristine membrane. In order to have further insight into the role of fillers, diffusion and solubility measurements were carried out. The results showed that the presence of highly porous fillers resulted in increasing the diffusion coefficient while the solubility showed a slight drop. The RTIL filled membranes showed higher CO2/CH4 and CO2/N2 selectivity than unfilled membranes while the permeability dropped slightly. The increase in selectivity was due to the highly selective RTIL used in this work. The study revealed that RTIL filled mixed matrix membranes are an interesting candidate for gas separation membranes.

Keywords: ionic liquids, CO2 separation, membranes, mixed matrix membranes

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Authors: Carlos Amnael Orozco-Diaz, Robert David Moorehead, Gwendolen Reilly, Fiona Gilchrist, Cheryl Ann Miller

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The current gold-standard for maxillofacial reconstruction surgery (MRS) utilizes auto-grafted cancellous bone as a filler. This study was aimed towards developing a polylactic-acid/hydroxyapatite (PLA-HA) composite suitable for fused-deposition 3D printing. Functionalization of the polymer through the addition of HA was directed to promoting bone-regeneration properties so that the material can rival the performance of cancellous bone grafts in terms of bone-lesion repair. This kind of composite enables the production of MRS implants based off 3D-reconstructions from image studies – namely computed tomography – for anatomically-correct fitting. The present study encompassed in-vitro degradation and in-vitro biocompatibility profiling for 3D-printed PLA and PLA-HA composites. PLA filament (Verbatim Co.) and Captal S hydroxyapatite micro-scale HA powder (Plasma Biotal Ltd) were used to produce PLA-HA composites at 5, 10, and 20%-by-weight HA concentration. These were extruded into 3D-printing filament, and processed in a BFB-3000 3D-Printer (3D Systems Co.) into tensile specimens, and were mechanically challenged as per ASTM D638-03. Furthermore, tensile specimens were subjected to accelerated degradation in phosphate-buffered saline solution at 70°C for 23 days, as per ISO-10993-13-2010. This included monitoring of mass loss (through dry-weighing), crystallinity (through thermogravimetric analysis/differential thermal analysis), molecular weight (through gel-permeation chromatography), and tensile strength. In-vitro biocompatibility analysis included cell-viability and extracellular matrix deposition, which were performed both on flat surfaces and on 3D-constructs – both produced through 3D-printing. Discs of 1 cm in diameter and cubic 3D-meshes of 1 cm3 were 3D printed in PLA and PLA-HA composites (n = 6). The samples were seeded with 5000 MG-63 osteosarcoma-like cells, with cell viability extrapolated throughout 21 days via resazurin reduction assays. As evidence of osteogenicity, collagen and calcium deposition were indirectly estimated through Sirius Red staining and Alizarin Red staining respectively. Results have shown that 3D printed PLA loses structural integrity as early as the first day of accelerated degradation, which was significantly faster than the literature suggests. This was reflected in the loss of tensile strength down to untestable brittleness. During degradation, mass loss, molecular weight, and crystallinity behaved similarly to results found in similar studies for PLA. All composite versions and pure PLA were found to perform equivalent to tissue-culture plastic (TCP) in supporting the seeded-cell population. Significant differences (p = 0.05) were found on collagen deposition for higher HA concentrations, with composite samples performing better than pure PLA and TCP. Additionally, per-cell-calcium deposition on the 3D-meshes was significantly lower when comparing 3D-meshes to discs of the same material (p = 0.05). These results support the idea that 3D-printable PLA-HA composites are a viable resorbable material for artificial grafts for bone-regeneration. Degradation data suggests that 3D-printing of these materials – as opposed to other manufacturing methods – might result in faster resorption than currently-used PLA implants.

Keywords: bone regeneration implants, 3D-printing, in vitro testing, biocompatibility, polymer degradation, polymer-ceramic composites

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Authors: C. Labate, M. P. De Santo, G. Lombardo, R. Barberi, M. Lombardo, N. M. Ziebarth

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In the past decades, the importance of corneal biomechanics in the normal and pathological functions of the eye has gained its credibility. In fact, the mechanical properties of biological tissues are essential to their physiological function. We are convinced that an improved understanding of the nanomechanics of corneal tissue is important to understand the basic molecular interactions between collagen fibrils. Ultimately, this information will help in the development of new techniques to cure ocular diseases and in the development of biomimetic materials. Therefore, nanotechnology techniques are powerful tools and, in particular, Atomic Force Microscopy has demonstrated its ability to reliably characterize the biomechanics of biological tissues either at the micro- or nano-level. In the last years, we have investigated the mechanical anisotropy of the human corneal stroma at both the tissue and molecular levels. In particular, we have focused on corneal cross-linking, an established procedure aimed at slowing down or halting the progression of the disease known as keratoconus. We have obtained the first evidence that riboflavin/UV-A corneal cross-linking induces both an increase of the elastic response and a decrease of the viscous response of the most anterior stroma at the scale of stromal molecular interactions.

Keywords: atomic force spectroscopy, corneal stroma, cross-linking, viscoelasticity

Procedia PDF Downloads 312

Authors: Rahul Saraswat

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More recently, a focus has been given to replacing machined stainless steel metal flow fields with inexpensive wire mesh current collectors. The flow fields are based on simple woven wire mesh screens of various stainless steels, which are sandwiched between a thin metal plate of the same material to create a bipolar plate/flow field configuration for use in a stack. Major advantages of using stainless steel wire screens include the elimination of expensive raw materials as well as machining and/or other special fabrication costs. The objective of the project is to improve the performance of the passive direct methanol fuel cell without increasing the cost of the cell and to make it as compact and light as possible. From the literature survey, it was found that very little is done in this direction, and the following methodology was used. 1. The passive direct methanol fuel cell (DMFC) can be made more compact, lighter, and less costly by changing the material used in its construction. 2. Controlling the fuel diffusion rate through the cell improves the performance of the cell. A passive liquid feed direct methanol fuel cell (DMFC) was fabricated using a given MEA (Membrane Electrode Assembly) and tested for different current collector structures. Mesh current collectors of different mesh densities along with different support structures, were used, and the performance was found to be better. Methanol concentration was also varied. Optimisation of mesh size, support structure, and fuel concentration was achieved. Cost analysis was also performed hereby. From the performance analysis study of DMFC, we can conclude with the following points: Area specific resistance (ASR) of wire mesh current collectors is lower than the ASR of stainless steel current collectors. Also, the power produced by wire mesh current collectors is always more than that produced by stainless steel current collectors. 1. Low or moderate methanol concentrations should be used for better and stable DMFC performance. 2. Wiremesh is a good substitute for stainless steel for current collector plates of passive DMFC because of its lower cost (by about 27 %), flexibility, and light in weight characteristics of wire mesh.

Keywords: direct methanol fuel cell, membrane electrode assembly, mesh, mesh size, methanol concentration, support structure

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Authors: Arif İsmailov, Naila Hasanova, Gunay Orujalieva

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Homocysteine (HCY) is an indicator of prognostic value in monitoring regenerative processes in osteoporosis and osteoporotic fractures. The osteoporosis is known to be a serious health and economic problem, especially for women in the postmenopausal period. The study was carried out on patients 45-83 years old divided into 3 groups: group I – 14 patients with osteoporosis , group II – 15 patients with non-osteoporotic fractures, group III – 25 patients with osteoporotic fractures. The control group consisted of practically healthy 14 people. A blood sample was taken at 3 stages to monitor the dynamics of HCY level: on the 1st day before treatment, on the 10th day of treatment and 1 month after it. Blood levels of Hcy were determined at a wavelength of 450 nm by the ELİSA(Cloud Clone Corp.Elisa kits,USA). The statistical evaluation was performed by using SPSS 26.0 program (IBM SPSS Inc., USA).The results showed that on the 1st day before the treatment HCY concentration was statistically increased 2.7 times(PU = 0.108) in group I, 5.6 times (PU <0.001) in group II and 6.5 times (PU <0.001) in group III compared to the control group. Thus, the average value of HCY in group I was 1.76 ± 0.56 μg/ml; in group II – 3.57 ± 0.62 μg/ml; in group III – 4.2 ± 0.50 μg/ml. HCY level increases more sharply after fractures,especially in osteoporotic patients. In treatment period Vitamin D plays an important role in synthesis of the Cystathionine β‐synthase enzyme, which regulates HCY metabolism. Increased Hcy levels could lead to an increase in the risk of fracture through the interference in collagen cross-linking.

Keywords: homocysteine, osteoporosis, osteoporotic fractures, Vitamin D

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Authors: Abdel-Mohdy, A. Abou-Okeil, S. El-Sabagh, S. M. El-Sawy

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Chitosan polyacrylic acid composite membranes were prepared by a bulk polymerization method in the presence of N, N'-methylene bisacrylamide (crosslinker) and ammonium persulphate as initiator. Membranes prepared from this copolymer in presence and absence of Ag nanoparticles were characterized by measuring mechanical and physical properties, water up-take and antibacterial properties. The results obtained indicated that the prepared membranes have antibacterial properties which increases with adding Ag nanoparticles.

Keywords: Ag nanoparticles , antimicrobial, Membrane, composites, mechanical properties, physical properties

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Authors: Jyh-Ping Chen, Chia-Lin Sheu

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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

Procedia PDF Downloads 181

Authors: Atlal El-Assaad

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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

Procedia PDF Downloads 113

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*

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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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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

Procedia PDF Downloads 254

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

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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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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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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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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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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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Authors: Husain S. Yawer, Vasim Raja Panwar, Nidhi Priya

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The objective of this study is to evaluate and compare the role of nano-gelatin and Bioengineered Scaffolds on the attachment, proliferation, and osteogenic differentiation of human dental pulp stem cells (DPSCs). Tooth decay and early fall have each been one of the most prevailing dental disorders which cause physical and emotional suffering and compromise the patient's quality of life. The design of novel scaffolding materials will be based on mimicking the architecture of natural dental extracellular matrix which may provide as in vivo environments for proper cell growth. This methodology will involve the combination of nano-fibred gelatin as well as biodegradable hydrogel based tooth scaffold. We have measured and optimized the Dental Pulp Stem Cells growth profile in cultures carried out on collagen-coated plastic surface, however, for tissue regeneration study, we aim to develop an enhanced microenvironment for stem cell growth and dental tissue regeneration. We believe biomimetic cell adhesion and scaffolds might provide a near in vivo growth environment for proper growth and differentiation of human DPSCs, which further help in dentin/pulp tissue regeneration.

Keywords: nano-gelatin, stem cells, dental pulp, scaffold

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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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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

Procedia PDF Downloads 131

Authors: Alexander Vandesompele, Joni Dambre

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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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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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Authors: Lyanne Rodriguez, Eduardo Fuente, Andrés Trostchansky, Ivan Palomo

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Cardiovascular diseases (CVD) are the leading cause of death in the world. The development of platelet-rich thrombi has been considered a trigger for acute cardiovascular events. A healthy diet, rich in fruit and vegetables, has been related to increased protection against cardiovascular events. Previous studies have observed that tomato pomace has a potent antiplatelet activity, due could be attributed to its high content of fatty acids (> 30%). It has been shown that unsaturated fatty acids can undergo endogenous intracellular nitration reactions during digestion after lipid consumption. Additionally, nitrated fatty acids (NO2-FA) can significantly reduce atherosclerotic lesion formation, inhibiting the expression of adhesion molecules on dysfunctional endothelium and platelet activation. In this work, we have proposed the nitration of fatty acids present in tomato pomace to improve its antiplatelet action. The gastric digestion of the tomato pomace allowed the nitration of the fatty acids, while by HPLC/MS/MS we were able to identify and quantify the nitrated fatty acids. The nitrated tomase extracts showed antiplatelet potential when platelets were stimulated with TRAP-6 and collagen. This activity was related to the presence of nitrated linoleic acid, which inhibited platelet activation by flow cytometry. The knowledge about the antiplatelet activity of nitrated fatty acids from tomato pomace will further develop new and more effective agents.

Keywords: cardiovascular, tomato extracts, nitrated fatty acids, antiplatelet activity

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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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Authors: Aziz Ghoufi, Ayman Kanaan

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Clean water is ubiquitous from drinking to agriculture and from energy supply to industrial manufacturing. Since the conventional water sources are becoming increasingly rare, the development of new technologies for water supply is crucial to address the world’s clean water needs in the 21st century. Desalination is in many regards the most promising approach to long-term water supply since it potentially delivers an unlimited source of fresh water. Seawater desalination using reverse osmosis (RO) membranes has become over the past decade a standard approach to produce fresh water. While this technology has proven to be efficient, it remains however relatively costly in terms of energy input due to the use of high-pressure pumps resulting of the low water permeation through polymeric RO membranes. Recently, water channels incorporated in lipidic and polymeric membranes were demonstrated to provide a selective water translocation that enables to break permeability- selectivity trade-off. Biomimetic Artificial Water channels (AWCs) are becoming highly attractive systems to achieve a selective transport of water. The first developed AWCs formed from imidazole quartet (I-quartet) embedded in lipidic membranes exhibited an ion selectivity higher than AQPs however associated with a lower water flow performance. Recently it has been conducted pioneer work in this field with the fabrication of the first AWC@Polyamide(PA) composite membrane with outstanding desalination performance. However, the microscopic desalination mechanism in play is still unknown and its understanding represents the shortest way for a long-term conception and design of AWC@PA composite membranes with better performance. In this work we gain an unprecedented fundamental understanding and rationalization of the nanostructuration of the AWC@PA membranes and the microscopic mechanism at the origin of their water transport performance from advanced molecular simulations. Using osmotic molecular dynamics simulations and a non-equilibrium method with water slab control, we demonstrate an increase in porosity near the AWC@PA interfaces, enhancing water transport without compromising the rejection rate. Indeed, the water transport pathways exhibit a single-file structure connected by hydrogen bonds. Finally, by comparing AWC@PA and PA membranes, we show that the difference in water flux aligns well with experimental results, validating the model used.

Keywords: water desalination, biomimetic membranes, molecular simulation, nanochannels

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Authors: Faris M. Altaf, Abdullah M Elkeshy

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Forty-two albino male rats aged between 30 and 40 days (weighted 200 g to 250 g) were used in the present study. The left sural nerves of 36 rats were subjected to crush injury at 1 to 6 weeks intervals using 6 animals at each interval. The right and left sural nerves of the rest 6 rats were used as a control. After 2 weeks of the crush injury, the endoneurium showed channel-like spaces that were lined by the fibroblast-like cells and collagen bundles. These channels contained degenerated myelin and were connected with the perivascular and subperineurial spaces. Some of the flattened fibroblast-like cells were arranged in several layers in the subperineurial and perivascular spaces, forming barrier-like cellular sheets localizing the endoneurial edema in these spaces. Fibroblast-like cells also wrapped the regenerating nerve fibers by their branching cytoplasmic processes. At the end of the third week, the flattened fibroblasts formed nearly continuous sheets in the subperineurial and perivascular spaces. Macrophages were frequently noticed between these cellular barrier-like sheets and in the subperineurial and perivascular spaces. Conclusion: it could be concluded that the endoneurial fibroblast-like cells form barrier-like cellular sheets that localized the endoneurial edema in the subperineurial and perivascular spaces and create also the endoneurial channel-like spaces containing degenerated myelin and endoneurial edema helping the resolution of such edema.

Keywords: sural nerve, endoneurial fibroblast-like cells, endoneurial edema, barrier-like and channel-like spaces

Procedia PDF Downloads 343