Search results for: cells encapsulation
515 Sol-Gel Derived 58S Bioglass Substituted by Li and Mg: A Comparative Evaluation on in vitro Bioactivity, MC3T3 Proliferation and Antibacterial Efficiency
Authors: Amir Khaleghipour, Amirhossein Moghanian, Elhamalsadat Ghaffari
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Modified bioactive glass has been considered as a promising multifunctional candidate in bone repair and regeneration due to its attractive properties. The present study mainly aims to evaluate how the individual substitution of lithium (L-BG) and magnesium (M-BG) for calcium can affect the in vitro bioactivity of sol-gel derived substituted 58S bioactive glass (BG); and to present one composition in both of the 60SiO₂–(36-x)CaO–4P₂O₅–(x)Li₂O and 60SiO₂–(36-x)CaO–4P₂O₅–(x)MgO quaternary systems (where x= 0, 5, 10 mol.%) with improved biocompatibility, enhanced alkaline phosphatase (ALP) activity, and the most efficient antibacterial activity against methicillin-resistant Staphylococcus aureus bacteria. To address these aims, and study the effect of CaO/Li₂O and CaO/MgO substitution up to 10 mol % in 58S-BGs, the samples were characterized by X-ray diffraction, Fourier transform infrared spectroscopy, inductively coupled plasma atomic emission spectrometry and scanning electron microscopy after immersion in simulated body fluid up to 14 days. Results indicated that substitution of either CaO/ Li₂O and CaO/ MgO had a retarding effect on in vitro hydroxyapatite (HA) formation due to the lower supersaturation degree for nucleation of HA compared with 58s-BG. Meanwhile, magnesium had a more pronounced effect. The 3-(4, 5dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and alkaline phosphatase (ALP) assays showed that both substitutions of CaO/ Li₂O and CaO/ MgO up to 5mol % in 58s-BGs led to increased biocompatibility and stimulated proliferation of the pre-osteoblast MC3T3 cells with respect to the control. On the other hand, substitution of either Li or Mg for Ca in the 58s BG composition resulted in improved bactericidal efficiency against MRSA bacteria. Taken together, sample 58s-BG with 5 mol % CaO/Li₂O substitution (BG-5L) was considered as a multifunctional biomaterial in bone repair/regeneration with improved biocompatibility, enhanced ALP activity as well enhanced antibacterial efficiency against methicillin-resistant Staphylococcus aureus (MRSA) bacteria among all of the synthesized L-BGs and M-BGs.Keywords: alkaline, alkaline earth, bioactivity, biomedical applications, sol-gel processes
Procedia PDF Downloads 190514 Numerical Modeling and Prediction of Nanoscale Transport Phenomena in Vertically Aligned Carbon Nanotube Catalyst Layers by the Lattice Boltzmann Simulation
Authors: Seungho Shin, Keunwoo Choi, Ali Akbar, Sukkee Um
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In this study, the nanoscale transport properties and catalyst utilization of vertically aligned carbon nanotube (VACNT) catalyst layers are computationally predicted by the three-dimensional lattice Boltzmann simulation based on the quasi-random nanostructural model in pursuance of fuel cell catalyst performance improvement. A series of catalyst layers are randomly generated with statistical significance at the 95% confidence level to reflect the heterogeneity of the catalyst layer nanostructures. The nanoscale gas transport phenomena inside the catalyst layers are simulated by the D3Q19 (i.e., three-dimensional, 19 velocities) lattice Boltzmann method, and the corresponding mass transport characteristics are mathematically modeled in terms of structural properties. Considering the nanoscale reactant transport phenomena, a transport-based effective catalyst utilization factor is defined and statistically analyzed to determine the structure-transport influence on catalyst utilization. The tortuosity of the reactant mass transport path of VACNT catalyst layers is directly calculated from the streaklines. Subsequently, the corresponding effective mass diffusion coefficient is statistically predicted by applying the pre-estimated tortuosity factors to the Knudsen diffusion coefficient in the VACNT catalyst layers. The statistical estimation results clearly indicate that the morphological structures of VACNT catalyst layers reduce the tortuosity of reactant mass transport path when compared to conventional catalyst layer and significantly improve consequential effective mass diffusion coefficient of VACNT catalyst layer. Furthermore, catalyst utilization of the VACNT catalyst layer is substantially improved by enhanced mass diffusion and electric current paths despite the relatively poor interconnections of the ion transport paths.Keywords: Lattice Boltzmann method, nano transport phenomena, polymer electrolyte fuel cells, vertically aligned carbon nanotube
Procedia PDF Downloads 201513 Poly-ε-Caprolactone Nanofibers with Synthetic Growth Factor Enriched Liposomes as Controlled Drug Delivery System
Authors: Vera Sovkova, Andrea Mickova, Matej Buzgo, Karolina Vocetkova, Eva Filova, Evzen Amler
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PCL (poly-ε-caprolactone) nanofibrous scaffolds with adhered liposomes were prepared and tested as a possible drug delivery system for various synthetic growth factors. TGFβ, bFGF, and IGF-I have been shown to increase hMSC (human mesenchymal stem cells) proliferation and to induce hMSC differentiation. Functionalized PCL nanofibers were prepared with synthetic growth factors encapsulated in liposomes adhered to them in three different concentrations. Other samples contained PCL nanofibers with adhered, free synthetic growth factors. The synthetic growth factors free medium served as a control. The interaction of liposomes with the PCL nanofibers was visualized by SEM, and the release kinetics were determined by ELISA testing. The potential of liposomes, immobilized on the biodegradable scaffolds, as a delivery system for synthetic growth factors, and as a suitable system for MSCs adhesion, proliferation and differentiation in vitro was evaluated by MTS assay, dsDNA amount determination, confocal microscopy, flow cytometry and real-time PCR. The results showed that the growth factors adhered to the PCL nanofibers stimulated cell proliferation mainly up to day 11 and that subsequently their effect was lower. By contrast, the release of the lowest concentration of growth factors from liposomes resulted in gradual proliferation of MSCs throughout the experiment. Moreover, liposomes, as well as free growth factors, stimulated type II collagen production, which was confirmed by immunohistochemical staining using monoclonal antibody against type II collagen. The results of this study indicate that growth factors enriched liposomes adhered to surface of PCL nanofibers could be useful as a drug delivery instrument for application in short timescales, be combined with nanofiber scaffolds to promote local and persistent delivery while mimicking the local microenvironment. This work was supported by project LO1508 from the Ministry of Education, Youth and Sports of the Czech RepublicKeywords: drug delivery, growth factors, hMSC, liposomes, nanofibres
Procedia PDF Downloads 290512 Preparedness for Microbial Forensics Evidence Collection on Best Practice
Authors: Victor Ananth Paramananth, Rashid Muniginin, Mahaya Abd Rahman, Siti Afifah Ismail
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Safety issues, scene protection, and appropriate evidence collection must be handled in any bio crime scene. There will be a scene or multi-scene to be cordoned for investigation in any bio-incident or bio crime event. Evidence collection is critical in determining the type of microbial or toxin, its lethality, and its source. As a consequence, from the start of the investigation, a proper sampling method is required. The most significant challenges for the crime scene officer would be deciding where to obtain samples, the best sampling method, and the sample sizes needed. Since there could be evidence in liquid, viscous, or powder shape at a crime scene, crime scene officers have difficulty determining which tools to use for sampling. To maximize sample collection, the appropriate tools for sampling methods are necessary. This study aims to assist the crime scene officer in collecting liquid, viscous, and powder biological samples in sufficient quantity while preserving sample quality. Observational tests on sample collection using liquid, viscous, and powder samples for adequate quantity and sample quality were performed using UV light in this research. The density of the light emission varies upon the method of collection and sample types. The best tools for collecting sufficient amounts of liquid, viscous, and powdered samples can be identified by observing UV light. Instead of active microorganisms, the invisible powder is used to assess sufficient sample collection during a crime scene investigation using various collection tools. The liquid, powdered and viscous samples collected using different tools were analyzed using Fourier transform infrared - attenuate total reflection (FTIR-ATR). FTIR spectroscopy is commonly used for rapid discrimination, classification, and identification of intact microbial cells. The liquid, viscous and powdered samples collected using various tools have been successfully observed using UV light. Furthermore, FTIR-ATR analysis showed that collected samples are sufficient in quantity while preserving their quality.Keywords: biological sample, crime scene, collection tool, UV light, forensic
Procedia PDF Downloads 195511 Cedrela Toona Roxb.: An Exploratory Study Describing Its Antidiabetic Property
Authors: Kinjal H. Shah, Piyush M. Patel
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Diabetes mellitus is considered to be a serious endocrine syndrome. Synthetic hypoglycemic agents can produce serious side effects including hematological effects, coma, and disturbances of the liver and kidney. In addition, they are not suitable for use during pregnancy. In recent years, there have been relatively few reports of short-term side effects or toxicity due to sulphonylureas. Published figures and frequency of side effects in large series of patient range from about 1 to 5%, with symptoms severe enough to lead to the withdrawal of the drug in less than 1 to 2%. Adverse effects, in general, have been of the following type: allergic skin reactions, gastrointestinal disturbances, blood dyscrasias, hepatic dysfunction, and hypoglycemia. The associated disadvantages with insulin and oral hypoglycemic agents have led to stimulation in the research for locating natural resources showing antidiabetic activity and to explore the possibilities of using traditional medicines with proper chemical and pharmacological profiles. Literature survey reveals that the inhabitants of Abbottabad district of Pakistan use the dried leaf powder along with table salt and water orally for treating diabetes, skin allergy, wounds and as a blood purifier, where they pronounced the plant locally as ‘Nem.' The detailed phytochemical investigation of the Cedrela toona Roxb. leaves for antidiabetic activity has not been documented. Hence, there is a need for phytochemical investigation of the leaves for antidiabetic activity. The collection of fresh leaves and authentification followed by successive extraction, phytochemical screening, and testing of antidiabetic activity. The blood glucose level was reduced maximum in ethanol extract at 5th and 7th h after treatment. Blood glucose was depressed by 8.2% and 10.06% in alloxan – induced diabetic rats after treatment which was comparable to the standard drug, Glibenclamide. This may be due to the activation of the existing pancreatic cells in diabetic rats by the ethanolic extract.Keywords: antidiabetic, Cedrela toona Roxb., phytochemical screening, blood glucose
Procedia PDF Downloads 260510 Optimization of Polymerase Chain Reaction Condition to Amplify Exon 9 of PIK3CA Gene in Preventing False Positive Detection Caused by Pseudogene Existence in Breast Cancer
Authors: Dina Athariah, Desriani Desriani, Bugi Ratno Budiarto, Abinawanto Abinawanto, Dwi Wulandari
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Breast cancer is a regulated by many genes. Defect in PIK3CA gene especially at position of exon 9 (E542K and E545K), called hot spot mutation induce early transformation of breast cells. The early detection of breast cancer based on mutation profile of this hot spot region would be hampered by the existence of pseudogene, marked by its substitution mutation at base 1658 (E545A) and deletion at 1659 that have been previously proven in several cancers. To the best of the authors’ knowledge, until recently no studies have been reported about pseudogene phenomenon in breast cancer. Here, we reported PCR optimization to to obtain true exon 9 of PIK3CA gene from its pseudogene hence increasing the validity of data. Material and methods: two genomic DNA with Dev and En code were used in this experiment. Two pairs of primer were design for Standard PCR method. The size of PCR products for each primer is 200bp and 400bp. While other primer was designed for Nested-PCR followed with DNA sequencing method. For Nested-PCR, we optimized the annealing temperature in first and second run of PCR, and the PCR cycle for first run PCR (15x versus 25x). Result: standard PCR using both primer pairs designed is failed to detect the true PIK3CA gene, appearing a substitution mutation at 1658 and deletion at 1659 of PCR product in sequence chromatogram indicated pseudogene. Meanwhile, Nested-PCR with optimum condition (annealing temperature for the first round at 55oC, annealing temperatung for the second round at 60,7oC with 15x PCR cycles) and could detect the true PIK3CA gene. Dev sample were identified as WT while En sample contain one substitution mutation at position 545 of exon 9, indicating amino acid changing from E to K. For the conclusion, pseudogene also exists in breast cancer and the apllication of optimazed Nested-PCR in this study could detect the true exon 9 of PIK3CA gene.Keywords: breast cancer, exon 9, hotspot mutation, PIK3CA, pseudogene
Procedia PDF Downloads 244509 Performance Assessment of Carrier Aggregation-Based Indoor Mobile Networks
Authors: Viktor R. Stoynov, Zlatka V. Valkova-Jarvis
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The intelligent management and optimisation of radio resource technologies will lead to a considerable improvement in the overall performance in Next Generation Networks (NGNs). Carrier Aggregation (CA) technology, also known as Spectrum Aggregation, enables more efficient use of the available spectrum by combining multiple Component Carriers (CCs) in a virtual wideband channel. LTE-A (Long Term Evolution–Advanced) CA technology can combine multiple adjacent or separate CCs in the same band or in different bands. In this way, increased data rates and dynamic load balancing can be achieved, resulting in a more reliable and efficient operation of mobile networks and the enabling of high bandwidth mobile services. In this paper, several distinct CA deployment strategies for the utilisation of spectrum bands are compared in indoor-outdoor scenarios, simulated via the recently-developed Realistic Indoor Environment Generator (RIEG). We analyse the performance of the User Equipment (UE) by integrating the average throughput, the level of fairness of radio resource allocation, and other parameters, into one summative assessment termed a Comparative Factor (CF). In addition, comparison of non-CA and CA indoor mobile networks is carried out under different load conditions: varying numbers and positions of UEs. The experimental results demonstrate that the CA technology can improve network performance, especially in the case of indoor scenarios. Additionally, we show that an increase of carrier frequency does not necessarily lead to improved CF values, due to high wall-penetration losses. The performance of users under bad-channel conditions, often located in the periphery of the cells, can be improved by intelligent CA location. Furthermore, a combination of such a deployment and effective radio resource allocation management with respect to user-fairness plays a crucial role in improving the performance of LTE-A networks.Keywords: comparative factor, carrier aggregation, indoor mobile network, resource allocation
Procedia PDF Downloads 178508 Non-Candida Albicans Candida: Virulence Factors and Species Identification in India
Authors: Satender Saraswat, Dharmendra Prasad Singh, Rajesh Kumar Verma, Swati Sarswat
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Background and Purpose: The predominant cause of candidiasis was Candida albicans which has shifted towards non-Candida albicans Candida (NCAC) (Candida species other than the C. albicans). NCAC, earlier considered non-pathogenic or minimally virulent, are now considered a primary cause of morbidity and mortality in immunocompromised. With the NCAC spp. gaining weightage in the clinical cases, this study was conducted to determine the prevalence of NCAC spp. in different clinical specimens and to assess a few of their virulence factors. Material and Methods: Routine samples for bacterial culture and sensitivity, showing colony characteristics like Candida on Blood Agar and microscopic features resembling Candida spp. were processed further. Candida isolates were tested for chlamydospore formation, biochemical tests including sugar fermentation and sugar assimilation tests, and growth at 42oC, colony colour on HiCrome™ Candida Differential Agar, HiCandida Identification Kit and VITEK-2 Compact. Virulence factors like adherence to buccal epithelial cells (ABEC), biofilm formation, hemolytic activity, and production of coagulase enzyme were also tested. Results: Mean age of the patients was 38.46 with a male-female ratio of 1.36:1. 137 Candida isolates were recovered. 45.3% isolates were isolated from urine, 19.7% from vaginal swabs and 13.9% from oropharyngeal swabs. 55 (40.1%) isolates of C. albicans and 82 (59.9%) of NCAC spp. were identified, with C. tropicalis (23.4%) in NCAC. C. albicans (3; 50%) was the commonest species in cases of candidemia. Haemolysin production (85.5%) and ABEC (78.2%) were the major virulence factors in C. albicans. C. tropicalis (59.4%) and C. dubliniensis (50%) showed maximum ABEC. Biofilm forming capacity was higher in C. tropicalis (78.1%) than C. albicans (67%). Conclusion: This study suggests varied prevalence and virulence based on geographical locations, even within a subcontinent. It clearly demarcates the emergence of NCAC and their predominance in different body fluids. Identification of Candida to species level should become a routine in all the laboratories.Keywords: ABEC, NCAC, non-Candida albicans Candida, Vitek-2TM compact
Procedia PDF Downloads 133507 Flow Reproduction Using Vortex Particle Methods for Wake Buffeting Analysis of Bluff Structures
Authors: Samir Chawdhury, Guido Morgenthal
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The paper presents a novel extension of Vortex Particle Methods (VPM) where the study aims to reproduce a template simulation of complex flow field that is generated from impulsively started flow past an upstream bluff body at certain Reynolds number Re-Vibration of a structural system under upstream wake flow is often considered its governing design criteria. Therefore, the attention is given in this study especially for the reproduction of wake flow simulation. The basic methodology for the implementation of the flow reproduction requires the downstream velocity sampling from the template flow simulation; therefore, at particular distances from the upstream section the instantaneous velocity components are sampled using a series of square sampling-cells arranged vertically where each of the cell contains four velocity sampling points at its corner. Since the grid free Lagrangian VPM algorithm discretises vorticity on particle elements, the method requires transformation of the velocity components into vortex circulation, and finally the simulation of the reproduction of the template flow field by seeding these vortex circulations or particles into a free stream flow. It is noteworthy that the vortex particles have to be released into the free stream exactly at same rate of velocity sampling. Studies have been done, specifically, in terms of different sampling rates and velocity sampling positions to find their effects on flow reproduction quality. The quality assessments are mainly done, using a downstream flow monitoring profile, by comparing the characteristic wind flow profiles using several statistical turbulence measures. Additionally, the comparisons are performed using velocity time histories, snapshots of the flow fields, and the vibration of a downstream bluff section by performing wake buffeting analyses of the section under the original and reproduced wake flows. Convergence study is performed for the validation of the method. The study also describes the possibilities how to achieve flow reproductions with less computational effort.Keywords: vortex particle method, wake flow, flow reproduction, wake buffeting analysis
Procedia PDF Downloads 311506 Multifunctional Plasmonic Ag-TiO2 Nano-biocompoistes: Surface Enhanced Raman Scattering and Anti-microbial Properties
Authors: Jai Prakash, Promod Kumar, Chantel Swart, J. H. Neethling, A. Janse van Vuuren, H. C. Swart
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Ag nanoparticles (NPs) have been used as functional nanomaterials due to their optical and antibacterial properties. Similarly, TiO2 photocatalysts have also been used as suitable nanomaterials for killing cancer cells, viruses and bacteria. Here, we report on multifunctional plasmonic Ag-TiO2 nano-biocomposite synthesized by the sol-gel technique and their optical, surface enhanced Raman scattering (SERS) and antibacterial activities. The as-prepared composites of Ag–TiO2 with different silver content and TiO2 nanopowder were characterized by X-ray diffraction, scanning electron microscopy, high-resolution transmission electron microscopy, energy-dispersed X-ray analysis (EDX), UV-vis and Raman spectroscopy. The Ag NPs were found to be uniformly distributed and strongly attached to the TiO2 matrix. The novel optical response of the Ag-TiO2 nanocomposites is due to the strong electric field from the surface plasmon excitation of the Ag NPs. The Raman spectrum of Ag-TiO2 nanocomposite was found to be enhanced as compared to TiO2. The enhancement of the low frequency band is evident. This indicates the SERS effect of the TiO2 NPs in close vicinity of Ag NPs. In addition, nanocomposites showed enhancement in the SERS signals of methyl orange (MO) dye molecules with increasing Ag content. The localized electromagnetic field from the surface plasmon excitation of the Ag NPs was responsible for the SERS signals of the TiO2 NPs and MO molecules. The antimicrobial effect of the Ag–TiO2 nanocomposites with different silver content and TiO2 nanopowder were carried out against the bacterium Staphylococcus aureus. The Ag–TiO2 composites showed antibacterial activity towards S. aureus with increasing Ag content as compared to the TiO2 nanopowder. These results foresee promising applications of the functional plasmonic metal−semiconductor based nanobiocomposites for both chemical and biological samples.Keywords: metal-Semiconductor, nano-Biocomposites, anti-microbial activity, surface enhanced Raman scattering
Procedia PDF Downloads 230505 Microfabrication of Three-Dimensional SU-8 Structures Using Positive SPR Photoresist as a Sacrificial Layer for Integration of Microfluidic Components on Biosensors
Authors: Su Yin Chiam, Qing Xin Zhang, Jaehoon Chung
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Complementary metal-oxide-semiconductor (CMOS) integrated circuits (ICs) have obtained increased attention in the biosensor community because CMOS technology provides cost-effective and high-performance signal processing at a mass-production level. In order to supply biological samples and reagents effectively to the sensing elements, there are increasing demands for seamless integration of microfluidic components on the fabricated CMOS wafers by post-processing. Although the PDMS microfluidic channels replicated from separately prepared silicon mold can be typically aligned and bonded onto the CMOS wafers, it remains challenging owing the inherently limited aligning accuracy ( > ± 10 μm) between the two layers. Here we present a new post-processing method to create three-dimensional microfluidic components using two different polarities of photoresists, an epoxy-based negative SU-8 photoresist and positive SPR220-7 photoresist. The positive photoresist serves as a sacrificial layer and the negative photoresist was utilized as a structural material to generate three-dimensional structures. Because both photoresists are patterned using a standard photolithography technology, the dimensions of the structures can be effectively controlled as well as the alignment accuracy, moreover, is dramatically improved (< ± 2 μm) and appropriately can be adopted as an alternative post-processing method. To validate the proposed processing method, we applied this technique to build cell-trapping structures. The SU8 photoresist was mainly used to generate structures and the SPR photoresist was used as a sacrificial layer to generate sub-channel in the SU8, allowing fluid to pass through. The sub-channel generated by etching the sacrificial layer works as a cell-capturing site. The well-controlled dimensions enabled single-cell capturing on each site and high-accuracy alignment made cells trapped exactly on the sensing units of CMOS biosensors.Keywords: SU-8, microfluidic, MEMS, microfabrication
Procedia PDF Downloads 522504 Synthesis and Characterization of Graphene Composites with Application for Sustainable Energy
Authors: Daniel F. Sava, Anton Ficai, Bogdan S. Vasile, Georgeta Voicu, Ecaterina Andronescu
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The energy crisis and environmental contamination are very serious problems, therefore searching for better and sustainable renewable energy is a must. It is predicted that the global energy demand will double until 2050. Solar water splitting and photocatalysis are considered as one of the solutions to these issues. The use of oxide semiconductors for solar water splitting and photocatalysis started in 1972 with the experiments of Fujishima and Honda on TiO2 electrodes. Since then, the evolution of nanoscience and characterization methods leads to a better control of size, shape and properties of materials. Although the past decade advancements are astonishing, for these applications the properties have to be controlled at a much finer level, allowing the control of charge-carrier lives, energy level positions, charge trapping centers, etc. Graphene has attracted a lot of attention, since its discovery in 2004, due to the excellent electrical, optical, mechanical and thermal properties that it possesses. These properties make it an ideal support for photocatalysts, thus graphene composites with oxide semiconductors are of great interest. We present in this work the synthesis and characterization of graphene-related materials and oxide semiconductors and their different composites. These materials can be used in constructing devices for different applications (batteries, water splitting devices, solar cells, etc), thus showing their application flexibility. The synthesized materials are different morphologies and sizes of TiO2, ZnO and Fe2O3 that are obtained through hydrothermal, sol-gel methods and graphene oxide which is synthesized through a modified Hummer method and reduced with different agents. Graphene oxide and the reduced form could also be used as a single material for transparent conductive films. The obtained single materials and composites were characterized through several methods: XRD, SEM, TEM, IR spectroscopy, RAMAN, XPS and BET adsorption/desorption isotherms. From the results, we see the variation of the properties with the variation of synthesis parameters, size and morphology of the particles.Keywords: composites, graphene, hydrothermal, renewable energy
Procedia PDF Downloads 498503 Effect of Serine/Threonine Kinases on Autophagy Mechanism
Authors: Ozlem Oral, Seval Kilic, Ozlem Yedier, Serap Dokmeci, Devrim Gozuacik
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Autophagy is a degradation pathway, activating under stress conditions. It digests macromolecules, such as abnormal proteins and long-lived organelles by engulfing them and by subsequent delivery of the cargo to lysosomes. The members of the phospholipid-dependent serine/threonine kinases, involved in many signaling pathways, which are necessary for the regulation of cellular metabolic activation. Previous studies implicate that, serine/threonine kinases have crucial roles in the mechanism of many diseases depend on the activated and/or inactivated signaling pathway. Data indicates, the signaling pathways activated by serine/threonine kinases are also involved in activation of autophagy mechanism. However, the information about the effect of serine/threonine kinases on autophagy mechanism and the roles of these effects in disease formation is limited. In this study, we investigated the effect of activated serine/threonine kinases on autophagic pathway. We performed a commonly used autophagy technique, GFP-LC3 dot formation and by using microscopy analyses, we evaluated promotion and/or inhibition of autophagy in serine/threonine kinase-overexpressed fibroblasts as well as cancer cells. In addition, we carried out confocal microscopy analyses and examined autophagic flux by utilizing the differential pH sensitivities of RFP and GFP in mRFP-GFP-LC3 probe. Based on the shRNA-library based screening, we identified autophagy-related proteins affected by serine/threonine kinases. We further studied the involvement of serine/threonine kinases on the molecular mechanism of newly identified autophagy proteins and found that, autophagic pathway is indirectly controlled by serine/threonine kinases via specific autophagic proteins. Our data indicate the molecular connection between two critical cellular mechanisms, which have important roles in the formation of many disease pathologies, particularly cancer. This project is supported by TUBITAK-1001-Scientific and Technological Research Projects Funding Program, Project No: 114Z836.Keywords: autophagy, GFP-LC3 dot formation assay, serine/threonine kinases, shRNA-library screening
Procedia PDF Downloads 292502 Evaluation of κ -Carrageenan Hydrogel Efficiency in Wound-Healing
Authors: Ali Ayatic, Emad Mozaffari, Bahareh Tanhaei, Maryam Khajenoori, Saeedeh Movaghar Khoshkho, Ali Ayati
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The abuse of antibiotics, such as tetracycline (TC), is a great global threat to people and the use of topical antibiotics is a promising tact that can help to solve this problem. Antibiotic therapy is often appropriate and necessary for acute wound infections, while topical tetracycline can be highly efficient in improving the wound healing process in diabetics. Due to the advantages of drug-loaded hydrogels as wound dressing, such as ease of handling, high moisture resistance, excellent biocompatibility, and the ability to activate immune cells to speed wound healing, it was found as an ideal wound treatment. In this work, the tetracycline-loaded hydrogels combining agar (AG) and κ-carrageenan (k-CAR) as polymer materials were prepared, in which span60 surfactant was introduced inside as a drug carrier. The Field Emission Scanning Electron Microscopes (FESEM) and Fourier-transform infrared spectroscopy (FTIR) techniques were employed to provide detailed information on the morphology, composition, and structure of fabricated drug-loaded hydrogels and their mechanical properties, and hydrogel permeability to water vapor was investigated as well. Two types of gram-negative and gram-positive bacteria were used to explore the antibacterial properties of prepared tetracycline-contained hydrogels. Their swelling and drug release behavior was studied using the changing factors such as the ratio of polysaccharides (MAG/MCAR), the span60 surfactant concentration, potassium chloride (KCl) concentration and different release media (deionized water (DW), phosphate-buffered saline (PBS), and simulated wound fluid (SWF)) at different times. Finally, the kinetic behavior of hydrogel swelling was studied. Also, the experimental data of TC release to DW, PBS, and SWF using various mathematical models such as Higuchi, Korsmeyer-Peppas, zero-order, and first-order in the linear and nonlinear modes were evaluated.Keywords: drug release, hydrogel, tetracycline, wound healing
Procedia PDF Downloads 80501 Pathogenic Candida Biofilms Producers Involved in Healthcare Associated Infections
Authors: Ouassila Bekkal Brikci Benhabib, Zahia Boucherit Otmani, Kebir Boucherit, A. Seghir
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The establishment of intravenous catheters in hospitalized patient is an act common in many clinical situations. These therapeutic tools, from their insertion in the body, represent gateways including fungal germs prone. The latter can generate the growth of biofilms, which can be the cause of fungal infection. Faced with this problem, we conducted a study at the University Hospital of Tlemcen in the neurosurgery unit and aims to isolate and identify Candida yeasts from intravenous catheters. Then test their ability to form biofilms. Materials and methods: 256 patient hospitalized in surgery of the hospital in west Algeria were submitted to this study. All samples were taken from peripheral venous catheters implanted for 72 hours or more days. A total of 31 isolates of Candida species were isolated. MIC and SMIC are determined at 80% inhibition by the test XTT tetrazolium measured at 490 nm. The final concentrations of antifungal agent being between 0.03 and 16 mg / ml for amphotericin B and from 0.015 to 8 mg / mL caspofungin. Results: 31 Candida species isolates from catheters including 14 Candida albicans and 17 Candida non albicans . 21 strains of all the isolates were able to form biofilms. In their form of Planktonic cells, all isolates are 100% susceptible to antifungal agents tested. However, in their state of biofilms, more isolates have become tolerant to the tested antifungals. Conclusion: Candida yeasts isolated from intravascular catheters are considered an important virulence factor in the pathogenesis of infections. Their involvement in catheter-related infections can be disastrous for their potential to generate biofilms. They survive high concentrations of antifungal where treatment failure. Pending the development of a therapeutic approach antibiofilm related to catheters, their mastery is going through: -The risk of infection prevention based on the training and awareness of medical staff, -Strict hygiene and maximum asepsis, and -The choice of material limiting microbial colonization.Keywords: candida, biofilm, hospital, infection, amphotericin B, caspofungin
Procedia PDF Downloads 323500 Pioglitazone Ameliorates Methotrexate-Induced Renal Endothelial Dysfunction via Amending Detrimental Changes in Antioxidant Profile, Systemic Cytokines and Apoptotic Factors
Authors: Sahar M. El-Gowilly, Mai M. Helmy, Hanan M. El-Gowelli
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Methotrexate (MTX) is widely used in treatment of cancers and autoimmune diseases. However, nephrotoxicity is one of the most important side effects of MTX. The peroxisome proliferator-activated receptor gamma agonist, pioglitazone (PIO), is known to exert anti-inflammatory and reno-protective effects in various kidney injuries. The purpose of this study was to investigate the potential involvement of endothelial damage in MTX-induced renal injury and to elaborate the possible protective effect of PIO against MTX-induced nephropathy. Compared with saline-treated rats, treatment with MTX (7 mg/kg for 3 day) caused significant elevations in serum levels of urea and creatinine, increased renal nitrate/nitrite level and impaired renovascular responsiveness of isolated perfused kidney to endothelium-dependent vasodilations induced by acetylcholine (0.01-2.43 nmol) and isoprenaline (1µmol). These effects were abolished by concurrent treatment with PIO (2.5 mg/kg, for 5 days starting two days before MTX). Alternatively, MTX treatment did not affect endothelium-independent renovascular relaxation induced by sodium nitroprusside (1-30 μmole). The possibility that alterations in renal antioxidants, circulating cytokine and apoptotic factor (Fas) levels contributed to MTX-PIO interaction was assessed. PIO treatment abrogated renal oxidative stress (decreased reduced glutathione and catalase activity and increased malondialdehyde), elevated serum cytokine (interleukin-6, interleukin-10, tumor necrosis factor-alpha and transforming growth factor-beta1) and Fas induced by MTX. Histologically, MTX caused defused tubular cells swelling and vacuolization associated with endothelial damage in renal arterioles. These effects disappeared upon co-treated with PIO. Collectively, PIO abolished MTX-induced endothelium dysfunction and nephrotoxicity via ameliorating oxidative stress and rectifying cytokines and Fas abnormalities caused by MTX.Keywords: methotrexate, pioglitazone, endothelium, kidney
Procedia PDF Downloads 312499 Study of the Hysteretic I-V Characteristics in a Polystyrene/ZnO-Nanorods Stack Layer
Authors: You-Lin Wu, Yi-Hsing Sung, Shih-Hung Lin, Jing-Jenn Lin
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Performance improvement in optoelectronic devices such as solar cells and photodetectors has been reported when a polymer/ZnO nanorods stack is used. Resistance switching of polymer/ZnO nanocrystals (or nanorods) hybrid has also gained a lot of research interests recently. It has been reported that high- and low-resistance states of a metal/insulator/metal (MIM) structure diode with a polystyrene (PS) and ZnO hybrid as the insulator layer can be switched by applied bias after a high-voltage forming process, while the same device structure merely with a PS layer does not show any forming behavior. In this work, we investigated the current-voltage (I-V) characteristics of an MIM device with a PS/ZnO nanorods stack deposited on fluorine-doped tin oxide (FTO) glass substrate. The ZnO nanorods were grown by a hydrothermal method using a mixture of zinc nitrate, hexamethylenetetramine, and DI water. Following that, a PS layer was deposited by spin coating. Finally, the device with a structure of Ti/ PS/ZnO nanorods/FTO was completed by e-gun evaporated Ti layer on top of the PS layer. Semiconductor parameters analyzer Agilent 4156C was then used to measure the I-V characteristics of the device by applying linear ramp sweep voltage with sweep sequence of 0V → 4V → 0V → 3V → 0V → 2V → 0V → 1V → 0V in both positive and negative directions. It is interesting to find that the I-V characteristics are bias dependent and hysteretic, indicating that the device Ti/PS/ZnO nanorods/FTO structure has ferroelectricity. Our results also show that the maximum hysteresis loop height of the I-V characteristics as well as the voltage at which the maximum hysteresis loop height of each scan occurs increase with increasing maximum sweep voltage. It should be noticed that, although ferroelectricity has been found in ZnO at its melting temperature (1975℃) and in Li- or Co-doped ZnO, neither PS nor ZnO has ferroelectricity at room temperature. Using the same structure but with a PS or ZnO layer only as the insulator does not give and hysteretic I-V characteristics. It is believed that a charge polarization layer is induced near the PS/ZnO nanorods stack interface and thus causes the ferroelectricity in the device with Ti/PS/ZnO nanorods/FTO structure. Our results show that the PS/ZnO stack can find a potential application in a resistive switching memory device with MIM structure.Keywords: ferroelectricity, hysteresis, polystyrene, resistance switching, ZnO nanorods
Procedia PDF Downloads 311498 DNA-Polycation Condensation by Coarse-Grained Molecular Dynamics
Authors: Titus A. Beu
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Many modern gene-delivery protocols rely on condensed complexes of DNA with polycations to introduce the genetic payload into cells by endocytosis. In particular, polyethyleneimine (PEI) stands out by a high buffering capacity (enabling the efficient condensation of DNA) and relatively simple fabrication. Realistic computational studies can offer essential insights into the formation process of DNA-PEI polyplexes, providing hints on efficient designs and engineering routes. We present comprehensive computational investigations of solvated PEI and DNA-PEI polyplexes involving calculations at three levels: ab initio, all-atom (AA), and coarse-grained (CG) molecular mechanics. In the first stage, we developed a rigorous AA CHARMM (Chemistry at Harvard Macromolecular Mechanics) force field (FF) for PEI on the basis of accurate ab initio calculations on protonated model pentamers. We validated this atomistic FF by matching the results of extensive molecular dynamics (MD) simulations of structural and dynamical properties of PEI with experimental data. In a second stage, we developed a CG MARTINI FF for PEI by Boltzmann inversion techniques from bead-based probability distributions obtained from AA simulations and ensuring an optimal match between the AA and CG structural and dynamical properties. In a third stage, we combined the developed CG FF for PEI with the standard MARTINI FF for DNA and performed comprehensive CG simulations of DNA-PEI complex formation and condensation. Various technical aspects which are crucial for the realistic modeling of DNA-PEI polyplexes, such as options of treating electrostatics and the relevance of polarizable water models, are discussed in detail. Massive CG simulations (with up to 500 000 beads) shed light on the mechanism and provide time scales for DNA polyplex formation independence of PEI chain size and protonation pattern. The DNA-PEI condensation mechanism is shown to primarily rely on the formation of DNA bundles, rather than by changes of the DNA-strand curvature. The gained insights are expected to be of significant help for designing effective gene-delivery applications.Keywords: DNA condensation, gene-delivery, polyethylene-imine, molecular dynamics.
Procedia PDF Downloads 120497 Pioglitazone Ameliorates Methotrexate-Induced Renal Endothelial Dysfunction via Amending Detrimental Changes in Antioxidant Profile, Systemic Cytokines and Fas Production
Authors: Sahar M. El-Gowilly, Mai M. Helmy, Hanan M. El-Gowelli
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Methotrexate (MTX) is widely used in treatment of cancers and autoimmune diseases. However, nephrotoxicity is one of its most important side effects. The peroxisome proliferator-activated receptor gamma agonist, pioglitazone, is known to exert antiinflammatory and reno-protective effects in various kidney injuries. The purpose of this study was to investigate the potential involvement of endothelial damage in MTX-induced renal injury and to elaborate the possible protective effect of pioglitazone against MTX-induced endothelial impairment. Compared with saline-treated rats, treatment with MTX (7 mg/kg for 3 day) caused significant elevations in serum levels of urea and creatinine, increased renal nitrate/nitrite level and impaired renovascular responsiveness of isolated perfused kidney to endothelium-dependent vasodilations induced by acetylcholine (0.01-2.43 nmol) and isoprenaline (1µmol). These effects were abolished by concurrent treatment with pioglitazone (2.5 mg/kg, for 5 days starting two days before MTX). Alternatively, MTX treatment did not affect endothelium-independent renovascular relaxation induced by sodium nitroprusside (0.001-10 μmole). The possibility that alterations in renal antioxidants, circulating cytokine and apoptotic factor (Fas) levels contributed to MTX-pioglitazone interaction was assessed. Pioglitazone treatment abrogated renal oxidative stress (decreased reduced glutathione and catalase activity and increased malondialdehyde), elevated serum cytokine (interleukin-6, interleukin-10, tumor necrosis factor-alpha and transforming growth factor-beta1) and Fas induced by MTX. Histologically, MTX caused defused tubular cells swelling and vacuolization associated with endothelial damage in renal arterioles. These effects disappeared upon co-treated with pioglitazone. Collectively, pioglitazone abolished MTX-induced endothelium dysfunction and nephrotoxicity via ameliorating oxidative stress and rectifying cytokines and Fas abnormalities caused by MTX.Keywords: methotrexate, pioglitazone, endothelium, kidney
Procedia PDF Downloads 500496 A Review on Bioremediation of Waste Effluent Associated with Pulp and Paper Industry
Authors: Adamu Muhammed Tukur
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Pulp and paper industry is one of the fastest growing industries due to an increased demand in paper products. For it to satisfy this ever increasing demand, it adopts new technological innovations some of which are proved to affect our environment negatively. Global consumption of paper has increased by 400% in the last four decades and this suggests that more research is required to assess the impact of industrial effluents to our environment and public health. Paper products are generally biodegradable, however, the processes involved in its production which involve the use of mainly bleaching agents and other non-biodegradable substances pose serious problem to the environment. There are more than 250 chemicals released in paper mill waste and some are xenobiotics. Different methods such as physical and chemical methods can be adopted for the remediation of the effluents but are proved to be costly and not safe to the environment. On the other hand, biological method is shown to be less costly and environmentally friendly. Microorganisms and their enzymes have shown a promising future for bioremediation of effluents related to paper mill. Many studies prove that one of the major pollutants in the paper mill effluent is phenol especially its chlorinated derivatives. Pentachlorophenol is extremely hazardous to living cells and therefore need to be removed from the environment. Microorganisms including bacteria and fungi have the potential to degrade phenolic compounds e.g. Bacillus stearothermiphilus, Pseudomonas putida, Coricus versicolor, Sphingomonas chlorophenolica, Fusarium sp, Bacillus subtilis and P. aeroginosa. Enzymes used for the degradation include phenol hydrooxylase, polyphenoloxylase, laccase, peroxidase among others. Lignin is another important pollutant and is resistant to microbial degradation but it has been proved that certain bacteria and fungi like can degrade it. Among the fungi white-rot fungi like Fomes lividus and Trametes vesicolor are the most important bioremediators. This review focused on use of microorganism to reduce or eradicate pollutants released from the paper industry. It can serve as a review for further research to be conducted especially in the field of Biotechnology.Keywords: bioremediation, pulp and paper, pentachlorophenol, environment
Procedia PDF Downloads 326495 Collagen Hydrogels Cross-Linked by Squaric Acid
Authors: Joanna Skopinska-Wisniewska, Anna Bajek, Marta Ziegler-Borowska, Alina Sionkowska
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Hydrogels are a class of materials widely used in medicine for many years. Proteins, such as collagen, due to the presence of a large number of functional groups are easily wettable by polar solvents and can create hydrogels. The supramolecular network capable to swelling is created by cross-linking of the biopolymers using various reagents. Many cross-linking agents has been tested for last years, however, researchers still are looking for a new, more secure reactants. Squaric acid, 3,4-dihydroxy 3-cyclobutene 1,2- dione, is a very strong acid, which possess flat and rigid structure. Due to the presence of two carboxyl groups the squaric acid willingly reacts with amino groups of collagen. The main purpose of this study was to investigate the influence of addition of squaric acid on the chemical, physical and biological properties of collagen materials. The collagen type I was extracted from rat tail tendons and 1% solution in 0.1M acetic acid was prepared. The samples were cross-linked by the addition of 5%, 10% and 20% of squaric acid. The mixtures of all reagents were incubated 30 min on magnetic stirrer and then dialyzed against deionized water. The FTIR spectra show that the collagen structure is not changed by cross-linking by squaric acid. Although the mechanical properties of the collagen material deteriorate, the temperature of thermal denaturation of collagen increases after cross-linking, what indicates that the protein network was created. The lyophilized collagen gels exhibit porous structure and the pore size decreases with the higher addition of squaric acid. Also the swelling ability is lower after the cross-linking. The in vitro study demonstrates that the materials are attractive for 3T3 cells. The addition of squaric acid causes formation of cross-ling bonds in the collagen materials and the transparent, stiff hydrogels are obtained. The changes of physicochemical properties of the material are typical for cross-linking process, except mechanical properties – it requires further experiments. However, the results let us to conclude that squaric acid is a suitable cross-linker for protein materials for medicine and tissue engineering.Keywords: collagen, squaric acid, cross-linking, hydrogel
Procedia PDF Downloads 388494 Management of Autoimmune Diseases with Ayurveda
Authors: Simmi Chopra
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In the last few years, there has been a surge of Autoimmune diseases that have become more like an epidemic all over the world. The reasons vary from stress, insufficient sleep, smoking, genetics, environmental pollution, adulterated foods, and a diet full of “the deadly white,” which is white sugar and white flour. Most of the people diagnosed with these diseases are given steroids, opioids, supplements, or elimination diets to manage their lives, but most of them continue suffering to varying degrees. On the other hand, Ayurveda can help manage autoimmune problems effectively. Ayurveda is a 5000 years old holistic medical system from India that has an individualistic approach where health problems are looked at from the lens of balancing body and mind and by targeting the root cause of the problem. A combination of diet and lifestyle according to Ayurvedic principles, Ayurvedic herbal formulations and Ayurvedic therapies can help in the management of autoimmune and other chronic diseases. Panchkarma, which is an intense six weeks detox method, helps balance our body and mind, and has been very effective in managing autoimmune problems. The paper will introduce the basic concepts of Ayurveda and describe the terminologies- doshas, agni and ama. The paper will discuss the importance of diet and lifestyle according to the individual’s imbalance in the three functional parameters - doshas, which govern every aspect of our body and mind, our cells and tissues. The significance of agni, which can be correlated to digestive strength and ama, which can be correlated to toxins that are formed in our body leading to health problems, will be outlined. The Ayurvedic pathophysiology of autoimmune diseases will be discussed with emphasis on Rheumatoid arthritis, Multiple sclerosis and Psoriasis. Ayurvedic management will be discussed for these autoimmune conditions. As Ayurveda is an individualistic system, one protocol will not work for everyone. Therefore, case studies with Ayurvedic protocols for the above autoimmune disease will be presented. Conclusion: Ayurveda can help in managing as well as arresting the progression of autoimmune problems. Ayurveda is an ancient medical system, is much more needed today than ever. It is a tried and tested holistic system which has been practiced for the past many generations in India.Keywords: ayurveda, autoimmune, diseases, nutrition
Procedia PDF Downloads 66493 In vivo Antidiabetic and Antioxidant Potential of Pseudovaria macrophylla Extract
Authors: Aditya Arya, Hairin Taha, Ataul Karim Khan, Nayiar Shahid, Hapipah Mohd Ali, Mustafa Ali Mohd
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This study has investigated the antidiabetic and antioxidant potential of Pseudovaria macrophylla bark extract on streptozotocin–nicotinamide induced type 2 diabetic rats. LCMS-QTOF and NMR experiments were done to determine the chemical composition in the methanolic bark extract. For in vivo experiments, the STZ (60 mg/kg/b.w, 15 min after 120 mg/kg/1 nicotinamide, i.p.) induced diabetic rats were treated with methanolic extract of Pseuduvaria macrophylla (200 and 400 mg/kg∙bw) and glibenclamide (2.5 mg/kg) as positive control respectively. Biochemical parameters were assayed in the blood samples of all groups of rats. The pro-inflammatory cytokines, antioxidant status and plasma transforming growth factor βeta-1 (TGF-β1) were evaluated. The histological study of the pancreas was examined and its expression level of insulin was observed by immunohistochemistry. In addition, the expression of glucose transporters (GLUT 1, 2 and 4) were assessed in pancreas tissue by western blot analysis. The outcomes of the study displayed that the bark methanol extract of Pseuduvaria macrophylla has potentially normalized the elevated blood glucose levels and improved serum insulin and C-peptide levels with significant increase in the antioxidant enzyme, reduced glutathione (GSH) and decrease in the level of lipid peroxidation (LPO). Additionally, the extract has markedly decreased the levels of serum pro-inflammatory cytokines and transforming growth factor beta-1 (TGF-β1). Histopathology analysis demonstrated that Pseuduvaria macrophylla has the potential to protect the pancreas of diabetic rats against peroxidation damage by downregulating oxidative stress and elevated hyperglycaemia. Furthermore, the expression of insulin protein, GLUT-1, GLUT-2 and GLUT-4 in pancreatic cells was enhanced. The findings of this study support the anti-diabetic claims of Pseudovaria macrophylla bark.Keywords: diabetes mellitus, Pseuduvaria macrophylla, alkaloids, caffeic acid
Procedia PDF Downloads 357492 Highly Active, Non-Platinum Metal Catalyst Material as Bi-Functional Air Cathode in Zinc Air Battery
Authors: Thirupathi Thippani, Kothandaraman Ramanujam
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Current research on energy storage has been paid to metal-air batteries, because of attractive alternate energy source for the future. Metal – air batteries have the probability to significantly increase the power density, decrease the cost of energy storage and also used for a long time due to its high energy density, low-level pollution, light weight. The performance of these batteries mostly restricted by the slow kinetics of the oxygen reduction reaction (ORR) and oxygen evolution reaction (OER) on cathode during battery discharge and charge. The ORR and OER are conventionally carried out with precious metals (such as Pt) and metal oxides (such as RuO₂ and IrO₂) as catalysts separately. However, these metal-based catalysts are regularly undergoing some difficulties, including high cost, low selectivity, poor stability and unfavorable to environmental effects. So, in order to develop the active, stable, corrosion resistance and inexpensive bi-functional catalyst material is mandatory for the commercialization of zinc-air rechargeable battery technology. We have attempted and synthesized non-precious metal (NPM) catalysts comprising cobalt and N-doped multiwalled carbon nanotubes (N-MWCNTs-Co) were synthesized by the solid-state pyrolysis (SSP) of melamine with Co₃O₄. N-MWCNTs-Co acts as an excellent electrocatalyst for both the oxygen reduction reaction (ORR) and the oxygen evolution reaction (OER), and hence can be used in secondary metal-air batteries and in unitized regenerative fuel cells. It is important to study the OER and ORR at high concentrations of KOH as most of the metal-air batteries employ KOH concentrations > 4M. In the first 16 cycles of the zinc-air battery while using N-MWCNTs-Co, 20 wt.% Pt/C or 20 wt.% IrO₂/C as air electrodes. In the ORR regime (the discharge profile of the zinc-air battery), the cell voltage exhibited by N-MWCNTs-Co was 44 and 83 mV higher (based on 5th cycle) in comparison to of 20 wt.% Pt/C and 20 wt.% IrO₂/C respectively. To demonstrate this promise, a zinc-air battery was assembled and tested at a current density of 0.5 Ag⁻¹ for charge-discharge 100 cycles.Keywords: oxygen reduction reaction (ORR), oxygen evolution reaction(OER), non-platinum, zinc air battery
Procedia PDF Downloads 234491 Identification of the Antimicrobial Effect of Liquorice Extracts on Gram-Positive Bacteria: Determination of Minimum Inhibitory Concentration and Mechanism of Action Using a luxABCDE Reporter Strain
Authors: Madiha El Awamie, Catherine Rees
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Natural preservatives have been used as alternatives to traditional chemical preservatives; however, a limited number have been commercially developed and many remain to be investigated as sources of safer and effective antimicrobials. In this study, we have been investigating the antimicrobial activity of an extract of Glycyrrhiza glabra (liquorice) that was provided as a waste material from the production of liquorice flavourings for the food industry, and to investigate if this retained the expected antimicrobial activity so it could be used as a natural preservative. Antibacterial activity of liquorice extract was screened for evidence of growth inhibition against eight species of Gram-negative and Gram-positive bacteria, including Listeria monocytogenes, Listeria innocua, Staphylococcus aureus, Enterococcus faecalis and Bacillus subtilis. The Gram-negative bacteria tested include Pseudomonas aeruginosa, Escherichia coli and Salmonella typhimurium but none of these were affected by the extract. In contrast, for all of the Gram-positive bacteria tested, growth was inhibited as monitored using optical density. However parallel studies using viable count indicated that the cells were not killed meaning that the extract was bacteriostatic rather than bacteriocidal. The Minimum Inhibitory Concentration [MIC] and Minimum Bactericidal Concentration [MBC] of the extract was also determined and a concentration of 50 µg ml-1 was found to have a strong bacteriostatic effect on Gram-positive bacteria. Microscopic analysis indicated that there were changes in cell shape suggesting the cell wall was affected. In addition, the use of a reporter strain of Listeria transformed with the bioluminescence genes luxABCDE indicated that cell energy levels were reduced when treated with either 12.5 or 50 µg ml-1 of the extract, with the reduction in light output being proportional to the concentration of the extract used. Together these results suggest that the extract is inhibiting the growth of Gram-positive bacteria only by damaging the cell wall and/or membrane.Keywords: antibacterial activity, bioluminescence, Glycyrrhiza glabra, natural preservative
Procedia PDF Downloads 340490 Synthesis and Characterization of the Carbon Spheres Built Up from Reduced Graphene Oxide
Authors: Takahiro Saida, Takahiro Kogiso, Takahiro Maruyama
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The ordered structural carbon (OSC) material is expected to apply to the electrode of secondary batteries, the catalyst supports, and the biomaterials because it shows the low substance-diffusion resistance by its uniform pore size. In general, the OSC material is synthesized using the template material. Changing size and shape of this template provides the pore size of OSC material according to the purpose. Depositing the oxide nanosheets on the polymer sphere template by the layer by layer (LbL) method was reported as one of the preparation methods of OSC material. The LbL method can provide the controlling thickness of structural wall without the surface modification. When the preparation of the uniform carbon sphere prepared by the LbL method which composed of the graphene oxide wall and the polymethyl-methacrylate (PMMA) core, the reduction treatment will be the important object. Since the graphene oxide has poor electron conductivity due to forming a lot of functional groups on the surface, it could be hard to apply to the electrode of secondary batteries and the catalyst support of fuel cells. In this study, the graphene oxide wall of carbon sphere was reduced by the thermal treatment under the vacuum conditions, and its crystalline structure and electronic state were characterized. Scanning electron microscope images of the carbon sphere after the heat treatment at 300ºC showed maintaining sphere shape, but its shape was collapsed with increasing the heating temperature. In this time, the dissolution rate of PMMA core and the reduction rate of graphene oxide were proportionate to heating temperature. In contrast, extending the heating time was conducive to the conservation of the sphere shape. From results of X-ray photoelectron spectroscopy analysis, its electronic state of the surface was indicated mainly sp² carbon. From the above results, we succeeded in the synthesis of the sphere structure composed by the reduction graphene oxide.Keywords: carbon sphere, graphene oxide, reduction, layer by layer
Procedia PDF Downloads 141489 Multilayered Assembly of Gelatin on Nanofibrous Matrix for 3-D Cell Cultivation
Authors: Ji Un Shin, Wei Mao, Hyuk Sang Yoo
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Electrospinning is a versatile tool for fabricating nano-structured polymeric materials. Gelatin hydrogels are considered to be a good material for cell cultivation because of high water swellability as well as good biocompatibility. Three-dimensional (3-D) cell cultivation is a desirable method of cell cultivation for preparing tissues and organs because cell-to-cell interactions or cell-to-matrix interactions can be much enhanced through this approach. For this reason, hydrogels were widely employed as tissue scaffolds because they can support cultivating cells and tissue in multi-dimensions. Major disadvantages of hydrogel-based cell cultivation include low mechanical properties, lack of topography, which should be enhanced for successful tissue engineering. Herein we surface-immobilized gelatin on the surface of nanofibrous matrix for 3-D cell cultivation in topographical cues added environments. Electrospun nanofibers were electrospun with injection of poly(caprolactone) through a single nozzle syringe. Electrospun meshes were then chopped up with a high speed grinder to fine powders. This was hydrolyzed in optimized concentration of sodium hydroxide solution from 1 to 6 hours and harvested by centrifugation. The freeze-dried powders were examined by scanning electron microscopy (SEM) for revealing the morphology and fibrilar shaped with a length of ca. 20um was observed. This was subsequently immersed in gelatin solution for surface-coating of gelatin, where the process repeated up to 10 times for obtaining desirable coating of gelatin on the surface. Gelatin-coated nanofibrils showed high waterswellability in comparison to the unmodified nanofibrils, and this enabled good dispersion properties of the modified nanofibrils in aqueous phase. The degree of water-swellability was increased as the coating numbers of gelatin increased, however, it did not any meaning result after 10 times of gelatin coating process. Thus, by adjusting the gelatin coating times, we could successfully control the degree of hydrophilicity and water-swellability of nanofibrils.Keywords: nano, fiber, cell, tissue
Procedia PDF Downloads 167488 Localisation of Fluorescently Labelled Drug-Free Phospholipid Vesicles to the Cartilage Surface of Rat Synovial Joints
Authors: Sam Yurdakul, Nick Baverstock, Jim Mills
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TDT 064 (FLEXISEQ®) is a drug-free gel used to treat osteoarthritis (OA)-associated pain and joint stiffness. It contains ultra-deformable phospholipid Sequessome™ vesicles, which can pass through the skin barrier intact. In six randomized OA studies, topical TDT 064 was well tolerated and improved joint pain, physical function and stiffness. In the largest study, these TDT 064-mediated effects were statistically significantly greater than oral placebo and equivalent to celecoxib. To understand the therapeutic effects of TDT 064, we investigated the localisation of the drug-free vesicles within rat synovial joints. TDT 064 containing DiO-labelled Sequessome™ vesicles was applied to the knees of four 6-week-old CD® hairless rats (10 mg/kg/ joint), 2–3 times/day, for 3 days (representing the recommended clinical dose). Eighteen hours later, the animals and one untreated control were sacrificed, and the knee joints isolated, flash frozen and embedded in Acrytol Mounting Media™. Approximately 15 sections (10 µm) from each joint were analysed by fluorescence microscopy. To investigate whether the localisation of DiO fluorescence was associated with intact vesicles, an anti-PEG monoclonal antibody (mAb) was used to detect Tween, a constituent of Sequessome™ vesicles. Sections were visualized at 484 nm (DiO) and 647 nm (anti-PEG mAb) and analysed using inForm 1.4 (Perkin Elmer, Inc.). Significant fluorescence was observed at 484 nm in sections from TDT 064-treated animals. No non-specific fluorescence was observed in control sections. Fluorescence was detected as discrete vesicles on the cartilage surfaces, inside the cartilaginous matrix and within the synovial space. The number of DiO-labelled vesicles in multiple fields of view was consistent and >100 in sections from four different treated knees. DiO and anti-PEG mAb co-localised within the collagenous tissues in four different joint sections. Under higher magnification (40x), vesicles were seen in the intercellular spaces of the synovial joint tissue, but no fluorescence was seen inside cells. These data suggest that the phospholipid vesicles in TDT 064 localize at the surface of the joint cartilage; these vesicles may therefore be supplementing the phospholipid deficiency reported in OA and acting as a biolubricant within the synovial joint.Keywords: joint pain, osteoarthritis, phospholipid vesicles, TDT 064
Procedia PDF Downloads 443487 Large-Scale Screening for Membrane Protein Interactions Involved in Platelet-Monocyte Interactions
Authors: Yi Sun, George Ed Rainger, Steve P. Watson
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Background: Beyond the classical roles in haemostasis and thrombosis, platelets are important in the initiation and development of various thrombo-inflammatory diseases. In atherosclerosis and deep vein thrombosis, for example, platelets bridge monocytes with endothelium and form heterotypic aggregates with monocytes in the circulation. This can alter monocyte phenotype by inducing their activation, stimulating adhesion and migration. These interactions involve cell surface receptor-ligand pairs on both cells. This list is likely incomplete as new interactions of importance to platelet biology are continuing to be discovered as illustrated by our discovery of PEAR-1 binding to FcεR1α. Results: We have developed a highly sensitive avidity-based assay to identify novel extracellular interactions among 126 recombinantly-expressed platelet cell surface and secreted proteins involved in platelet aggregation. In this study, we will use this method to identify novel platelet-monocyte interactions. We aim to identify ligands for orphan receptors and novel partners of well-known proteins. Identified interactions will be studied in preliminary functional assays to demonstrate relevance to the inflammatory processes supporting atherogenesis. Conclusions: Platelet-monocyte interactions are essential for the development of thromboinflammatory disease. Up until relatively recently, technologies only allow us to limit our studies on each individual protein interaction at a single time. These studies propose for the first time to study the cell surface platelet-monocyte interactions in a systematic large-scale approach using a reliable screening method we have developed. If successful, this will likely to identify previously unknown ligands for important receptors that will be investigated in details and also provide a list of novel interactions for the field. This should stimulate studies on developing alternative therapeutic strategies to treat vascular inflammatory disorders such as atherosclerosis, DVT and sepsis and other clinically important inflammatory conditions.Keywords: membrane proteins, large-scale screening, platelets, recombinant expression
Procedia PDF Downloads 151486 Optimization of SOL-Gel Copper Oxide Layers for Field-Effect Transistors
Authors: Tomas Vincze, Michal Micjan, Milan Pavuk, Martin Weis
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In recent years, alternative materials are gaining attention to replace polycrystalline and amorphous silicon, which are a standard for low requirement devices, where silicon is unnecessarily and high cost. For that reason, metal oxides are envisioned as the new materials for these low-requirement applications such as sensors, solar cells, energy storage devices, or field-effect transistors. Their most common way of layer growth is sputtering; however, this is a high-cost fabrication method, and a more industry-suitable alternative is the sol-gel method. In this group of materials, many oxides exhibit a semiconductor-like behavior with sufficiently high mobility to be applied as transistors. The sol-gel method is a cost-effective deposition technique for semiconductor-based devices. Copper oxides, as p-type semiconductors with free charge mobility up to 1 cm2/Vs., are suitable replacements for poly-Si or a-Si:H devices. However, to reach the potential of silicon devices, a fine-tuning of material properties is needed. Here we focus on the optimization of the electrical parameters of copper oxide-based field-effect transistors by modification of precursor solvent (usually 2-methoxy ethanol). However, to achieve solubility and high-quality films, a better solvent is required. Since almost no solvents have both high dielectric constant and high boiling point, an alternative approach was proposed with blend solvents. By mixing isopropyl alcohol (IPA) and 2-methoxy ethanol (2ME) the precursor reached better solubility. The quality of the layers fabricated using mixed solutions was evaluated in accordance with the surface morphology and electrical properties. The IPA:2ME solution mixture reached optimum results for the weight ratio of 1:3. The cupric oxide layers for optimal mixture had the highest crystallinity and highest effective charge mobility.Keywords: copper oxide, field-effect transistor, semiconductor, sol-gel method
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