Search results for: cheap scaffolds

Authors: Kirill Filianin, Satu-Pia Reinikainen, Tuomo Sainio

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Frequent measurements of product steam quality create a data overload that becomes more and more difficult to handle. In the current study, plant history data with multiple variables was successfully treated by principal component analysis to detect abnormal process behavior, particularly, in copper solvent extraction. The multivariate model is based on the concentration levels of main process metals recorded by the industrial on-stream x-ray fluorescence analyzer. After mean-centering and normalization of concentration data set, two-dimensional multivariate model under principal component analysis algorithm was constructed. Normal operating conditions were defined through control limits that were assigned to squared score values on x-axis and to residual values on y-axis. 80 percent of the data set were taken as the training set and the multivariate model was tested with the remaining 20 percent of data. Model testing showed successful application of control limits to detect abnormal behavior of copper solvent extraction process as early warnings. Compared to the conventional techniques of analyzing one variable at a time, the proposed model allows to detect on-line a process failure using information from all process variables simultaneously. Complex industrial equipment combined with advanced mathematical tools may be used for on-line monitoring both of process streams’ composition and final product quality. Defining normal operating conditions of the process supports reliable decision making in a process control room. Thus, industrial x-ray fluorescence analyzers equipped with integrated data processing toolbox allows more flexibility in copper plant operation. The additional multivariate process control and monitoring procedures are recommended to apply separately for the major components and for the impurities. Principal component analysis may be utilized not only in control of major elements’ content in process streams, but also for continuous monitoring of plant feed. The proposed approach has a potential in on-line instrumentation providing fast, robust and cheap application with automation abilities.

Keywords: abnormal process behavior, failure detection, principal component analysis, solvent extraction

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Authors: Syed Asif Hassan, Tabrej Khan

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Tuberculosis (TB) caused by bacillus Mycobacterium tuberculosis (Mtb) continues to take a disturbing toll on human life and healthcare facility worldwide. The global burden of TB remains enormous. The alarming rise of multi-drug resistant strains of Mycobacterium tuberculosis calls for an increase in research efforts towards the development of new target specific therapeutics against diverse strains of M. tuberculosis. Therefore, the discovery of new molecular scaffolds targeting new drug sites should be a priority for a workable plan for fighting resistance in Mycobacterium tuberculosis (Mtb). Mtb non-acylated lipoprotein LprG (Rv1411c) has a Toll-like receptor 2 (TLR2) agonist actions that depend on its association with triacylated glycolipids binding specifically with the hydrophobic pocket of Mtb LprG lipoprotein. The detection of a glycolipid carrier function has important implications for the role of LprG in Mycobacterial physiology and virulence. Therefore, considering the pivotal role of glycolipids in mycobacterial physiology and host-pathogen interactions, designing competitive antagonist (chemotherapeutics) ligands that competitively bind to glycolipid binding domain in LprG lipoprotein, will lead to inhibition of tuberculosis infection in humans. In this study, a unified approach involving ligand-based virtual screening protocol USRCAT (Ultra Shape Recognition) software and molecular docking studies using Auto Dock Vina 1.1.2 using the X-ray crystal structure of Mtb LprG protein was implemented. The docking results were further confirmed by DSX (DrugScore eXtented), a robust program to evaluate the binding energy of ligands bound to the Ligand binding domain of the Mtb LprG lipoprotein. The ligand, which has the higher hypothetical affinity, also has greater negative value. Based on the USRCAT, Lipinski’s values and molecular docking results, [(2R)-2,3-di(hexadecanoyl oxy)propyl][(2S,3S,5S,6R)-3,4,5-trihydroxy-2,6-bis[[(2R,3S,4S,5R,6S)-3,4,5-trihydroxy-6 (hydroxymethyl)tetrahydropyran-2-yl]oxy]cyclohexyl] phosphate (XPX) was confirmed as a promising drug-like lead compound (antagonist) binding specifically to the hydrophobic domain of LprG protein with affinity greater than that of PIM2 (agonist of LprG protein) with a free binding energy of -9.98e+006 Kcal/mol and binding affinity of -132 Kcal/mol, respectively. A further, in vitro assay of this compound is required to establish its potency in inhibiting molecular evasion mechanism of MTB within the infected host macrophages. These results will certainly be helpful in future anti-TB drug discovery efforts against Multidrug-Resistance Tuberculosis (MDR-TB).

Keywords: antagonist, agonist, binding affinity, chemotherapeutics, drug-like, multi drug resistance tuberculosis (MDR-TB), RV1411c protein, toll-like receptor (TLR2)

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Authors: Mohammad Hossein Pazandeh, Monir Doudi, Sona Rostampour Yasouri

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—Results The prudent administration of antibiotics aims to avoid the side effects and the microbes' resistance to antibiotics. An approach developing methods of local administration of antibiotics is especially required for localized infections caused by bacterial colonization of medical devices or implant materials. Among the wide variety of materials used as drug delivery systems, bioactive glasses (BG) have large utilization in regenerative medicine . firstly, the production of bioactive glass/nickel oxide/tin dioxide nanocomposite using sol-gel method, and then, the controlled release of imipenem from the double metal oxide/bioactive glass nanocomposite, and finally, the investigation of the antibacterial property of the nanocomposite. against a number of implant-related infectious agents. In this study, BG/SnO2 and BG/NiO single systema with different metal oxide present and BG/NiO/SnO2 nanocomposites were synthesized by sol-gel as drug carriers for tetracycline and imepinem. These two antibiotics were widely used for osteomyelitis because of its favorable penetration and bactericidal effect on all the probable osteomyelitis pathogens. The antibacterial activity of synthesized samples were evaluated against Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa as bacteria model using disk diffusion method. The BG modification using metal oxides results to antibacterial property of samples containing metal oxide with highest efficiency for nancomposite. bioactivity of all samples was assessed by determining the surface morphology, structural and composition changes using scanning electron microscopy (SEM), FTIR and X-ray diffraction (XRD) spectroscopy, respectively, after soaking in simulated body fluid (SBF) for 28 days. The hydroxyapatite formation was clearly observed as a bioactivity measurement. Then, BG nanocomposite sample was loaded using two antibiotics, separately and their release profiles were studied. The BG nancomposite sample was shown the slow and continuous drug releasing for a period of 72 hours which is desirable for a drug delivery system. The loaded antibiotic nanocomposite sample retaining antibacterial property and showing inactivation effect against bacteria under test. The modified bioactive glass forming hydroxyapatite with controlled release drug and effective against bacterial infections can be introduced as scaffolds for bone implants after clinical trials for biomedical applications . Considering the formation of biofilm by infectious bacteria after sticking on the surfaces of implants, medical devices, etc. Also, considering the complications of traditional methods, solving the problems caused by the above-mentioned microorganisms in technical and biomedical industries was one of the necessities of this research.

Keywords: antibacterial, bioglass, drug delivery system, sol- gel

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Authors: Ahmed Auda, Manjree Agarwala, Giles Hardya, Yonglin Rena

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Botrytis cinerea is a major pest for many plants. It can attack a wide range of plant parts. It can attack buds, flowers, and leaves, stems, and fruit. However, B. cinerea can be mixed with other diseases that cause the same damage. There are many species of botrytis and more than one different strains of each. Botrytis might infect the foliage of nursery stock stored through winter in damp conditions. There are no known resistant plants. Botrytis must have nutrients or food source before it infests the plant. Nutrients leaking from wounded plant parts or dying tissue like old flower petals give the required nutrients. From this food, the fungus becomes more attackers and invades healthy tissue. Dark to light brown rot forms in the ill tissue. High humidity conditions support the growth of this fungus. However, we suppose that selection pressure can act on the morphological and neurophysiologic filter properties of the receiver and on both the biochemical and the physiological regulation of the signal. Communication is implied when signal and receiver evolves toward more and more specific matching, culminating. In other hand, receivers respond to portions of a body odor bouquet which is released to the environment not as an (intentional) signal but as an unavoidable consequence of metabolic activity or tissue damage. Each year Botrytis species can cause considerable economic losses to plant crops. Even with the application of strict quarantine and control measures, these fungi can still find their way into crops and cause the imposition of onerous restrictions on exports. Blueberry fruit mould caused by a fungal infection usually results in major losses during post-harvest storage. Therefore, the management of infection in early stages of disease development is necessary to minimize losses. The overall purpose of this study will develop sensitive, cheap, quick and robust diagnostic techniques for the detection of B. cinerea in blueberry. The specific aim was designed to investigate the performance of volatile organic compounds (VOCs) in the detection and discrimination of blueberry fruits infected by fungal pathogens with an emphasis on Botrytis in the early storage stage of post-harvest.

Keywords: botrytis cinerea, blueberry, GC/MS, VOCs

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Authors: Anshuman Srivastava, Shiv Singh, Sheelendra Pratap Singh

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A simple, sensitive and effective gas chromatography tandem mass spectrometry (GC-MS/MS) method was developed for simultaneous analysis of multi pesticide residues (organophosphate, organochlorines, synthetic pyrethroids and herbicides) in food commodities using phenolic resin based activated carbon fibers (ACFs) as reversed-dispersive solid phase extraction (r-DSPE) sorbent in modified QuEChERS (Quick Easy Cheap Effective Rugged Safe) method. The acetonitrile-based QuEChERS technique was used for the extraction of the analytes from food matrices followed by sample cleanup with ACFs instead of traditionally used primary secondary amine (PSA). Different physico-chemical characterization techniques such as Fourier transform infrared spectroscopy, scanning electron microscopy, X-ray diffraction and Brunauer-Emmet-Teller surface area analysis were employed to investigate the engineering and structural properties of ACFs. The recovery of pesticides and herbicides was tested at concentration levels of 0.02 and 0.2 mg/kg in different commodities such as cauliflower, cucumber, banana, apple, wheat and black gram. The recoveries of all twenty-six pesticides and herbicides were found in acceptable limit (70-120%) according to SANCO guideline with relative standard deviation value < 15%. The limit of detection and limit of quantification of the method was in the range of 0.38-3.69 ng/mL and 1.26 -12.19 ng/mL, respectively. In traditional QuEChERS method, PSA used as r-DSPE sorbent plays a vital role in sample clean-up process and demonstrates good recoveries for multiclass pesticides. This study reports that ACFs are better in terms of removal of co-extractives in comparison of PSA without compromising the recoveries of multi pesticides from food matrices. Further, ACF replaces the need of charcoal in addition to the PSA from traditional QuEChERS method which is used to remove pigments. The developed method will be cost effective because the ACFs are significantly cheaper than the PSA. So the proposed modified QuEChERS method is more robust, effective and has better sample cleanup efficiency for multiclass multi pesticide residues analysis in different food matrices such as vegetables, grains and fruits.

Keywords: QuEChERS, activated carbon fibers, primary secondary amine, pesticides, sample preparation, carbon nanomaterials

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Authors: S. Jayasinghe, W. G. D. Wickramasingha, V. Karunaratne, D. N. Karunaratne, A. Ekanayake

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Multi-drug resistant microbial pathogens are a serious global health problem, and hence, there is an urgent necessity for discovering new drug therapeutics. However, finding alternatives is a one of the biggest challenges faced by the global drug industry due to the spiraling high cost and serious side effects associated with modern medicine. On the other hand, plants and their secondary metabolites can be considered as good sources of scaffolds to provide structurally diverse bioactive compounds as potential therapeutic agents. 6β-hydroxy betunolic acid is a triterpenoid isolated from bark of Schumacheria castaneifolia which is an endemic plant to Sri Lanka which has shown antibacterial activity against both Staphylococcus aureus (ATCC 29213) and methicillin-resistant S. aureus with Minimum Inhibition Concentration (MIC) of 16 µg/ml. The objective of this study was to determine the anti-bacterial activity for the derivatives of 6β- hydroxy betunolic acid against standard strains of Staphylococcus aureus (ATCC 29213 and ATCC 25923), Enterococcus faecalis (ATCC 29212), Escherichia coli (ATCC 35218 and ATCC 25922), Pseudomonas aeruginosa (ATCC 27853), carbepenemas produce Kebsiella pneumonia (ATCC BAA 1705) and carbepenemas non produce Kebsiella pneumonia (ATCC BAA 1706) and four stains of clinically isolated methicillin resistance S. aureus and Acinetobacter. Structural analogues of 6β-hydroxy betunolic acid were synthesized by modifying the carbonyl group at C-3 to obtain olefin and oxime, the hydroxyl group at C-6 position to a ketone, the carboxylic acid at C-17 to obtain amide and halo ester and the olefin group at C-20 position to obtain epoxide. Chemical structures of the synthesized analogues were confirmed with spectroscopic data and antibacterial activity was determined through broth micro dilution assay. Results revealed that 6β- hydroxy betunolic acid shows significant antibacterial activity only against the Gram positive strains and it was inactive against all the tested Gram negative strains for the tested concentration range. However, structural modifications into oxime and olefin at C-3, ketone at C-6 and epoxide at C-20 decreased its antibacterial activity against the gram positive organisms and it was totally lost with the both modifications at C-17 into amide and ester. These results concluded that the antibacterial activity of 6β- hydroxy betunolic acid and derivatives is predominantly depending on the cell wall difference of the bacteria and the presence of carboxylic acid at C-17 is highly important for the antibacterial activity against Gram positive organisms.

Keywords: antibacterial activity, 6β- hydroxy betunolic acid, broth micro dilution assay, structure activity relationship

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Authors: Louise H. Gracioso, Marcela P.G. Baltazar, Ingrid R. Avanzi, Bruno Karolski, Luciana J. Gimenes, Claudio O. Nascimento, Elen A. Perpetuo

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Introduction: Higher copper concentrations promote a selection pressure on organisms such as plants, fungi and bacteria, which allows surviving only the resistant organisms to the contaminated site. This selective pressure keeps only the organisms most resistant to a specific condition and subsequently increases their bioremediation potential. Despite the bacteria importance for biosphere maintenance, it is estimated that only a small fraction living microbial species has been described and characterized. Due to the molecular biology development, tools based on analysis 16S ribosomal RNA or another specific gene are making a new scenario for the characterization studies and identification of microorganisms in the environment. News identification of microorganisms methods have also emerged like Biotyper (MALDI / TOF), this method mass spectrometry is subject to the recognition of spectroscopic patterns of conserved and features proteins for different microbial species. In view of this, this study aimed to isolate bacteria resistant to copper present in a Copper Processing Area (Sossego Mine, Canaan, PA) and identifies them in two different methods: Recent (spectrometry mass) and conventional. This work aimed to use them for a future bioremediation of this Mining. Material and Methods: Samples were collected at fifteen different sites of five periods of times. Microorganisms were isolated from mining wastes by culture enrichment technique; this procedure was repeated 4 times. The isolates were inoculated into MJS medium containing different concentrations of chloride copper (1mM, 2.5mM, 5mM, 7.5mM and 10 mM) and incubated in plates for 72 h at 28 ºC. These isolates were subjected to mass spectrometry identification methods (Biotyper – MALDI/TOF) and 16S gene sequencing. Results: A total of 105 strains were isolated in this area, bacterial identification by mass spectrometry method (MALDI/TOF) achieved 74% agreement with the conventional identification method (16S), 31% have been unsuccessful in MALDI-TOF and 2% did not obtain identification sequence the 16S. These results show that Biotyper can be a very useful tool in the identification of bacteria isolated from environmental samples, since it has a better value for money (cheap and simple sample preparation and MALDI plates are reusable). Furthermore, this technique is more rentable because it saves time and has a high performance (the mass spectra are compared to the database and it takes less than 2 minutes per sample).

Keywords: copper mining area, bioremediation, microorganisms, identification, MALDI/TOF, RNA 16S

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Authors: Alberto Mínguez-Martínez, Jesús De Vicente Y Oliva

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It seems that we live in a world in which the trend in industrial environments is the miniaturization of systems and materials and the fabrication of parts at the micro-and nano-scale. The problem arises when manufacturers want to study the quality of their production. This characteristic is becoming crucial due to the evolution of the industry and the development of Industry 4.0. As Industry 4.0 is based on digital models of production and processes, having accurate measurements becomes capital. At this point, the metrology field plays an important role as it is a powerful tool to ensure more stable production to reduce scrap and the cost of non-conformities. The most extended measuring instruments that allow us to carry out accurate measurements at these scales are optical microscopes, whether they are traditional, confocal, focus variation microscopes, profile projectors, or any other similar measurement system. However, the accuracy of measurements is connected to the traceability of them to the SI unit of length (the meter). The fact of providing adequate traceability to 2D and 3D dimensional measurements at micro-and nano-scale in industrial environments is a problem that is being studied, and it does not have a unique answer. In addition, if commercial material standards for micro-and nano-scale are considered, we can find that there are two main problems. On the one hand, those material standards that could be considered complete and very interesting do not give traceability of dimensional measurements and, on the other hand, their calibration is very expensive. This situation implies that these kinds of standards will not succeed in industrial environments and, as a result, they will work in the absence of traceability. To solve this problem in industrial environments, it becomes necessary to have material standards that are easy to use, agile, adaptive to different forms, cheap to manufacture and, of course, traceable to the definition of meter with simple methods. By using these ‘customized standards’, it would be possible to adapt and design measuring procedures for each application and manufacturers will work with some traceability. It is important to note that, despite the fact that this traceability is clearly incomplete, this situation is preferable to working in the absence of it. Recently, it has been demonstrated the versatility and the utility of using laser technology and other AM technologies to manufacture customized material standards. In this paper, the authors propose to manufacture a customized material standard using an ultraviolet laser system and a method to calibrate it. To conclude, the results of the calibration carried out in an accredited dimensional metrology laboratory are presented.

Keywords: industrial environment, material standards, optical measuring instrument, traceability

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Authors: Asim Abbasi, Muhammad Sufyan, Muhammad Kamran, Iqra

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Development of resistance in insect pests against various groups of insecticides has prompted the use of alternative integrated pest management approaches. Among these induced host plant resistance represents an important strategy as it offers a practical, cheap and long lasting solution to keep pests populations below economic threshold level (ETL). Silicon (Si) has a major role in regulating plant eco-relationship by providing strength to the plant in the form of anti-stress mechanism which was utilized in coping with the environmental extremes to get a better yield and quality end produce. Among biotic stresses, insect herbivore signifies one class against which Si provide defense. Silicon in its neutral form (H₄SiO₄) is absorbed by the plants via roots through an active process accompanied by the help of different transporters which were located in the plasma membrane of root cells or by a passive process mostly regulated by transpiration stream, which occurs via the xylem cells along with the water. Plants tissues mainly the epidermal cell walls are the sinks of absorbed silicon where it polymerizes in the form of amorphous silica or monosilicic acid. The noteworthy function of this absorbed silicon is to provide structural rigidity to the tissues and strength to the cell walls. Silicon has both direct and indirect effects on insect herbivores. Increased abrasiveness and hardness of epidermal plant tissues and reduced digestibility as a result of deposition of Si primarily as phytoliths within cuticle layer is now the most authenticated mechanisms of Si in enhancing plant resistance to insect herbivores. Moreover, increased Si content in the diet also impedes the efficiency by which insects transformed consumed food into the body mass. The palatability of food material has also been changed by Si application, and it also deters herbivore feeding for food. The production of defensive compounds of plants like silica and phenols have also been amplified by the exogenous application of silicon sources which results in reduction of the probing time of certain insects. Some studies also highlighted the role of silicon at the third trophic level as it also attracts natural enemies of insects attacking the crop. Hence, the inclusion of Si in pest management approaches can be a healthy and eco-friendly tool in future.

Keywords: defensive, phytoliths, resistance, stresses

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Authors: Jeet Chakraborty, Sanjay Dutta

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Antibiotic resistance by bacteria has proved to be a severe threat to mankind in recent times, and this fortifies an urgency to design and develop potent antibacterial small molecules/compounds with nonconventional mechanisms than the conventional ones. DNA carries the genetic signature of any organism, and bacteria maintain their genomic DNA inside the cell in a well-regulated compact form with the help of various nucleoid associated proteins like HU, HNS, etc. These proteins control various fundamental processes like gene expression, replication, etc., inside the cell. Alteration of the native DNA structure of bacteria can lead to severe consequences in cellular processes inside the bacterial cell that ultimately result in the death of the organism. The change in the global DNA structure by small molecules initiates a plethora of cellular responses that have not been very well investigated. Echinomycin and Triostin-A are biologically active Quinoxaline small molecules that typically consist of a quinoxaline chromophore attached with an octadepsipeptide ring. They bind to double-stranded DNA in a sequence-specific way and have high activity against a wide variety of bacteria, mainly against Gram-positive ones. To date, few synthetic quinoxaline scaffolds were synthesized, displaying antibacterial potential against a broad scale of pathogenic bacteria. QNOs (Quinoxaline N-oxides) are known to target DNA and instigate reactive oxygen species (ROS) production in bacteria, thereby exhibiting antibacterial properties. The divergent role of Quinoxaline small molecules in medicinal research qualifies them for the evaluation of their antimicrobial properties as a potential candidate. The previous study from our lab has given new insights on a 6-nitroquinoxaline derivative 1d as an intercalator of DNA, which induces conformational changes in DNA upon binding.7 The binding event observed was dependent on the presence of a crucial benzyl substituent on the quinoxaline moiety. This was associated with a large induced CD (ICD) appearing in a sigmoidal pattern upon the interaction of 1d with dsDNA. The induction of DNA superstructures by 1d at high Drug:DNA ratios was observed that ultimately led to DNA condensation. Eviction of invitro-assembled nucleosome upon treatment with a high dose of 1d was also observed. In this work, monoquinoxaline derivatives of 1d were synthesized by various modifications of the 1d scaffold. The set of synthesized 6-nitroquinoxaline derivatives along with 1d were all subjected to antibacterial evaluation across five different bacteria species. Among the compound set, 3a displayed potent antibacterial activity against Staphylococcus aureus bacteria. 3a was further subjected to various biophysical studies to check whether the DNA structural alteration potential was still intact. The biological response of S. aureus cells upon treatment with 3a was studied using various cell biology processes, which led to the conclusion that 3d can initiate DNA damage in the S. aureus cells. Finally, the potential of 3a in disrupting preformed S.aureus and S.epidermidis biofilms was also studied.

Keywords: DNA structural change, antibacterial, intercalator, DNA superstructures, biofilms

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Authors: Ayodeji Fasuyi

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Telfairia occidentalis is a leafy vegetable commonly grown in the tropics for nutritional benefits. The use of enzymes and probiotics is becoming prominent due to the ban on antibiotics as growth promoters in many parts of the world. It is conceived that with enzymes and probiotics additives, fibrous leafy vegetables can be incorporated into poultry feeds as protein source. However, certain antinutrients were also found in the leaves of Telfairia occidentalis. Four broiler starter and finisher diets were formulated for the two phases of the broiler experiments. A mixture of fiber degrading enzymes, Roxazyme G2 (combination of cellulase, glucanase and xylanase) and probiotics (Turbotox), a growth promoter, were used in broiler diets at 1:1. The Roxazyme G2/Turbotox mixtures were used in diets containing four varying levels of Telfairia occidentalis leaf meal (TOLM) at 0, 10, 20 and 30%. Diets 1 were standard broiler diets without TOLM and Roxazyme G2 and Turbotox additives. Diets 2, 3 and 4 had enzymes and probiotics additives. Certain mineral elements such as Ca, P, K, Na, Mg, Fe, Mn, Cu and Zn were found in notable quantities viz. 2.6 g/100 g, 1.2 g/100 g, 6.2 g/100 g, 5.1 g/100 g, 4.7 g/100 g, 5875 ppm, 182 ppm, 136 ppm and 1036 ppm, respectively. Phytin, phytin-P, oxalate, tannin and HCN were also found in ample quantities viz. 189.2 mg/100 g, 120.1 mg/100 g, 80.7 mg/100 g, 43.1 mg/100 g and 61.2 mg/100 g, respectively. The average weight gain was highest at 46.3 g/bird/day for birds on 10% TOLM diet but similar (P > 0.05) to 46.2 g/bird/day for birds on 20% TOLM. The feed conversion ratio (FCR) of 2.27 was the lowest and optimum for birds on 10% TOLM although similar (P > 0.05) to 2.29 obtained for birds on 20% TOLM. FCR of 2.61 was the highest at 2.61 for birds on 30% TOLM diet. The lowest FCR of 2.27 was obtained for birds on 10% TOLM diet although similar (P > 0.05) to 2.29 for birds on 20% TOLM diet. Most carcass characteristics and organ weights were similar (P > 0.05) for the experimental birds on the different diets except for kidney, gizzard and intestinal length. The values for kidney, gizzard and intestinal length were significantly higher (P < 0.05) for birds on the TOLM diets. The nitrogen retention had the highest value of 72.37 ± 0.10% for birds on 10% TOLM diet although similar (P > 0.05) to 71.54 ± 1.89 obtained for birds on the control diet without TOLM and enzymes/probiotics mixture. There was evidence of a better utilization of TOLM as a plant protein source. The carcass characteristics and organ weights all showed evidence of uniform tissue buildup and muscles development particularly in diets containing 10% of TOLM level. There was also better nitrogen utilization in birds on the 10% TOLM diet. Considering the cheap cost of TOLM, it is envisaged that its introduction into poultry feeds as a plant protein source will ultimately reduce the cost of poultry feeds.

Keywords: Telfairia occidentalis leaf meal, enzymes, probiotics, additives

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Authors: Dennis Otieno, Lilian Kirimi Nicholas Odhiambo, Hillary Bii

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Kenya is considered to be a middle level income country and usuaaly does not meet household food security needs especially in North and South eastern parts. Approximately half of the population is living under the poverty line (www, CIA 1, 2012). Agriculture is the largest sector in the country, employing 80% of the population. These are thereby directly dependent on the sufficiency of outputs received. This makes efficient, easy-accessible and cheap agricultural practices an important matter in order to improve food security. Maize is the prime staple food commodity in Kenya and represents a substantial share of people’s nutritional intake. This study is the result of questionnaire based interviews, Key informant and focus group discussion involving 220 small scale maize farmers Kenyan. The study was located to two separated areas; Lower Kuja, Bunyala, Nandi, Lower Nzoia, Perkerra, Mwea Bura, Hola and Galana Kulalu in Kenya. The questionnaire captured the farmers’ use and perceived importance of the use irrigation services and irrigated maize production. Viability was evaluated using the four indices which were all positive with NPV giving positive cash flows in less than 21 years at most for one season output. The mean willingness to pay was found to be KES 3082 and willingness to pay increased with increase in irrigation premiums. The economic value of water was found to be greater than the willingness to pay implying that irrigated maize production is sustainable. Farmers stated that viability was influenced by high output levels, good produce quality, crop of choice, availability of sufficient water and enforcement the last two factors had a positive influence while the other had negative effect on the viability of irrigated maize. A regression was made over the correlation between the willingness to pay for irrigated maize production using scheme and plot level factors. Farmers that already use other inputs such as animal manure, hired labor and chemical fertilizer should also have a demand for improved seeds according to Liebig's law of minimum and expansion path theory. The regression showed that premiums, and high yields have a positive effect on willingness to pay while produce quality, efficient fertilizer use, and crop season have a negative effect.

Keywords: maize, food security, profits, sustainability, willingness to pay

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Authors: Alexander A. Osmolovskiy, Anastasia V. Orekhova, Daria M. Bednenko, Yelyzaveta Boiko

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Micromycetes from Aspergillus genus can produce proteases capable of promoting proteolysis of hemostasis proteins or, along with hydrolytic activity, to show the ability to convert proenzymes of this system activating them into an active form. At the same time, practical medicine needs specific activators for quantitation of the level of some plasma enzymes, especially protein C and factor X, the lack of which leads to the development of thromboembolic diseases. Thus, some micromycetes of the genus Aspergillus were screened for the ability to synthesize extracellular proteases with promising activity for designing anti-thrombotic and diagnostic preparations. Such standard methods like salting out, electrophoresis, isoelectrofocusing were used for isolation, purification and study of physicochemical properties of proteases. Enzyme activity was measured spectrophotometrically fibrin as a substrate of the reaction and chromogenic peptide substrates of different proteases of the human hemostasis system. As a result of the screening, four active producers were selected: Aspergillus janus 301, A. flavus 1, A. terreus 2, and A. ochraceus L-1. The enzyme of A. janus 301 showed the greatest fibrinolytic activity (around 329.2 μmol Tyr/(ml × min)). The protease produced by A. terreus 2 had the highest plasmin-like activity (54.1 nmol pNA/(ml × min)), but fibrinolytic activity was lower than A. janus 301 demonstrated (25.2 μmol Tyr/(ml × min)). For extracellular protease of micromycete A. flavus a high plasmin-like activity was also shown (39.8 nmol pNA / (ml × min)). Moreover, according to our results proteases one of the fungi - A. terreus 2 were able to activate protein C of human plasma - the key factor of the human anticoagulant hemostasis system. This type of activity was 39.8 nmol pNA/(ml × min)). It was also shown that A. ochraceus L-1 could produce extracellular proteases with protein C and factor X activator activities (65.9 nmol pNA/(ml × min) and 34.6 nmol pNA/(ml × min) respectively). The maximum accumulation of the proteases falls on the 4th day of cultivation. Using isoelectrofocusing was demonstrated that the activation of both proenzymes might proceed via limited proteolysis induced by proteases of A. ochraceus L-1. The activatory activity of A. ochraceus L-1 proteases toward essential hemostatic proenzymes, protein C and X factor may be useful for practical needs. It is well known that similar enzymes, activators of protein C and X factor isolated from snake venom, South American copperhead Agkistrodon contortrix contortrix and Russell’s viper Daboia russelli russeli, respectively, are used for the in vitro diagnostics of the functional state of these proteins in blood plasma. Thus, the proteases of Aspergillus genus can be used as cheap components for enzyme thrombolytic preparations.

Keywords: anti-trombotic drugs, fibrinolysis, diagnostics, proteases, micromycetes

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Authors: M. Weller Jones, J. Moffat, J. Taylor, J. Hartland, M. Natarajan

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Background: Uganda’s teenage pregnancy rate remains a significant problem for female and maternal health in the country. Teenage pregnancy is linked to higher rates of maternal and neonatal mortality and morbidity, including preterm labour, obstructed labour, vesicovaginal fistulae, infections, and maternal mental health morbidity. In 2015, the National Strategy to End Child Marriage and Teenage Pregnancy was launched in Uganda. Research is needed so that the interventions in this Strategy can be effectively applied at a local level. This study at Kitovu and Villa Maria Hospitals, two local community hospitals near Masaka, Uganda, aimed to measure change in the local teenage pregnancy rate over the past 5 years; and to explore the awareness and attitudes of teenagers and healthcare professionals towards 1) teenage pregnancy and, 2) the challenges female adolescents still currently face. Method: Teenage delivery rate, type of delivery, incidence of complications in labour and neonatal and maternal outcomes were collected from the labour ward admission books, at both hospitals, for a six month time period in 2011 and 2016. At Kitovu Hospital, qualitative data regarding the experience of, and attitudes towards teenage pregnancy was collected from interviews conducted with 12 maternity staff members and with eight female teenagers, aged 16-19, who were pregnant or post-partum. Results: The proportion of total births to teenage mothers fell from 14% in 2011 to 7% in 2016 (Kitovu), but it remains higher in rural locations (19%, Villa Maria). Beliefs about exacerbating factors included: poor access to contraception; misconceptions that contraception is damaging to women’s health; failing sex education in schools; and poor awareness of national campaigns to reduce teenage pregnancy. Staff felt that the best way to tackle teenage pregnancy was to improve sex education in schools and to sensitise families to these issues. Six of the eight teenagers wanted more frequent sex education and easier, cheap access to contraception. Only one teenager saw positive consequences stating that teenage pregnancy would ‘avoid operations later in life.’ Discussion: Teenage pregnancy is a recognised problem and strategies in the Masaka region should focus on improving sex education in schools and initiating an organisation that educates and supplies free contraception to teenagers.

Keywords: adolescents, attitudes, teenage pregnancy, Uganda

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Authors: Kunal Bhardwaj, Christine Browne

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With the increase in global awareness of the environmental impacts of plastic-based products, there has been a massive drive to reduce our use of these products. Use of plastic-based substrates in electronic circuits has been a matter of concern recently. Plastics provide a very smooth and cheap surface for printing high-performance electronics due to their non-permeability to ink and easy mouldability. In this research, we explore the use of nano cellulose (NC) films in electronics as they provide an advantage of being 100% recyclable and eco-friendly. The main hindrance in the mass adoption of NC film as a substitute for plastic is its higher surface roughness which leads to ink penetration, and dispersion in the channels on the film. This research was conducted to tune the RMS roughness of NC films to a range where they can replace plastics in electronics(310-470nm). We studied the dependence of the surface roughness of the NC film on the following tunable aspects: 1) composition by weight of the NC suspension that is sprayed on a silicon wafer 2) the width and the depth of the channels on the silicon wafer used as a base. Various silicon wafers with channel depths ranging from 6 to 18 um and channel widths ranging from 5 to 500um were used as a base. Spray coating method for NC film production was used and two solutions namely, 1.5wt% NC and a 50-50 NC-CNC (cellulose nanocrystal) mixture in distilled water, were sprayed through a Wagner sprayer system model 117 at an angle of 90 degrees. The silicon wafer was kept on a conveyor moving at a velocity of 1.3+-0.1 cm/sec. Once the suspension was uniformly sprayed, the mould was left to dry in an oven at 50°C overnight. The images of the films were taken with the help of an optical profilometer, Olympus OLS 5000. These images were converted into a ‘.lext’ format and analyzed using Gwyddion, a data and image analysis software. Lowest measured RMS roughness of 291nm was with a 50-50 CNC-NC mixture, sprayed on a silicon wafer with a channel width of 5 µm and a channel depth of 12 µm. Surface roughness values of 320+-17nm were achieved at lower (5 to 10 µm) channel widths on a silicon wafer. This research opened the possibility of the usage of 100% recyclable NC films with an additive (50% CNC) in high-performance electronics. Possibility of using additives like Carboxymethyl Cellulose (CMC) is also being explored due to the hypothesis that CMC would reduce friction amongst fibers, which in turn would lead to better conformations amongst the NC fibers. CMC addition would thus be able to help tune the surface roughness of the NC film to an even greater extent in future.

Keywords: nano cellulose films, electronic circuits, nanocrystals and surface roughness

Procedia PDF Downloads 101

Authors: Amarachi Delight Onyemachi, Gregory Ikechukwu Onwuka

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The growing consumer trend for natural ingredients, functional foods with health benefits and the perceived risk of carcinogenesis associated with synthetic antioxidants have forced food manufacturers to look for alternatives for producing healthy and safe food. Herbs and spices are cheap, natural and harmless sources of antioxidants which can delay and prevent lipid oxidation of food products and also confer its unique organoleptic properties and health benefits to food products. The Nigerian climate has been proven to be conducive for the production of spices and herbs and is blessed bountifully with a wide range of them. Five selected Nigerian herbs and spices Piper guieense, Xylopia aethopica, Gongronema latifolium and Ocimum gratissimum were evaluated for their ability to act as radical scavengers. The spices were extracted with 80% ethanol and evaluated using total phenolic capacity (TPC), DPPH (1,1-diph diphenyl-2-picrylhydrazyl radical) ABTS (2,2’azinobis-(3-ethylbenzthiazoline-6-sulfonic acid)), total antioxidant capacity (TAC), reducing power (RP) assays. The TPC ranged from 5.33 µg GAE/mg (in Gongronema latifolium) to 15.55 µg GAE/mg (in Ocimum gratissimum). The DPPH and ABTS scavenging activity of the extracts ranged from 0.23-0.36 IC50 mg/ml and 2.32-7.25 Trolox equivalent % respectively. The TAC and RP of the extract ranged from 6.73-10.64 µg AAE/mg and 3.52-10.19 µg AAE/mg. The result of percentage yield of the extract ranged from as low as 9.94% in Gongronema latifolium and to as high as 23.85% in Xylopia aethopica. A very strong positive relationship existed between the total antioxidant capacity and total phenolic content of the tested herbs and spices (R2=0.96). All of the extracts exhibited different extent of strong antioxidant activity, high antioxidant activity was found in Ocimum gratissimum and Gongronema latifolium with the least. However, Gongronema latifolium possessed the highest total antioxidant capacity. These data confirm the appreciable antioxidant potentials and high phenolic content of Nigerian herbs and spices, thereby providing justification for their use in dishes and functional foods, prevention of cellular damage caused by free radicals and use as natural antioxidants in the food industry for prevention of lipid oxidation in food products. However, to utilize these natural antioxidants in food products, further analysis and studies of their behaviour in food systems at varying temperature, pH conditions and ionic concentrations should be carried out to displace the use of synthetic antioxidants like BHT and BHA.

Keywords: Antioxidant, free radicals, herbs, phenolic, spices

Procedia PDF Downloads 228

Authors: Juliana Kurniawati, Helen Diana Vida

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Visiting mall is one of the Indonesian communities’ lifestyle who live in urban areas. Indonesian people, especially who live in Jakarta, use a shopping mall as one of the favourite places to get pleasure. This mall visitors come from various social classes. They use the shopping mall as a place to identify themselves as urban people. Jakarta has a number of great shopping malls such as Plaza Indonesia, Plaza Senayan, Pondok Indah Mall, etc. The shopping malls become one of the popular places since Jakarta's public sphere such as parks and playgrounds are very limited in number compared to that of shopping malls. In Jakarta, people do not come to a shopping mall only for shopping. Sometimes they go there to look around, meet up with some friends, or watch a movie. We can find everything in the shopping malls. The principle of one-stop shopping becomes an attractive offer for urban people. The items for selling are various, from the cheap goods to the expensive ones. A new era in consumer culture began with the advent of shopping was localized in France in the 19th century. Since the development of the online store and the easier way to access the internet, everyone can shop 24 hours anywhere they want. The emergence of online store indirectly has an impact on the viability of conventional stores. In October 2017, in Indonesia, two outlets branded goods namely Lotus and Debenhams were closed. This may a result of increasingly rampant online stores and shopping style urban society shift. The rising of technology gives some influence on the development of e-commerce in Indonesia. Everyone can access e-commerce. However, those who can do it are the middle up class to high class people. The development of e-commerce in Indonesia is quite fast, we can observe the emergence of various online shopping sites on various social media platforms such as Zalora, Berrybenka, Bukalapak, Lazada, and Tokopedia. E-commerce is increasingly affecting people's lives in line with the development of lifestyle and increasing revenue. This research aims to know the reasons of urban society choosing e-commerce as a medium for grocery shopping, how e-commerce is affecting their shopping styles, as well as why society provides confidence in the online store for shopping. This research uses theories of lifestyle by David Chaney. The subject of this research is urban society who actively shop online on Zalora, the communities based in Jakarta. Zalora site was chosen because the site is selling branded goods. This research is expected to explain in detail about the changing style of the urban community from the shopping mall to digital media by emphasizing the aspect of public confidence towards the online store.

Keywords: e-commerce, shopping, lifestyle, changing

Procedia PDF Downloads 271

Authors: Rawan Ashraf, Ahmed E. Gomaa, Gehan Safwat, Ayman Diab

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Background: Three-dimensional (3D) bio-printing has become a versatile and powerful method for generating a variety of biological constructs, including bone or extracellular matrix scaffolds endo- or epithelial, muscle tissue, as well as organoids. Aim of the study: Fabricate a low cost DIY 3D bio-printer to produce 3D bio-printed products such as anti-microbial packaging or multi-organs on chips. We demonstrate the alignment between two types of 3D printer technology (3D Bio-printer and DLP) on Multi-organ-on-a-chip (multi-OoC) devices fabrication. Methods: First, Design and Fabrication of the Syringe Unit for Modification of an Off-the-Shelf 3D Printer, then Preparation of Hydrogel based on natural polymers Sodium Alginate and Gelatin, followed by acquisition of the cell suspension, then modeling the desired 3D structure. Preparation for 3D printing, then Cell-free and cell-laden hydrogels went through the printing process at room temperature under sterile conditions and finally post printing curing process and studying the printed structure regards physical and chemical characteristics. The hard scaffold of the Organ on chip devices was designed and fabricated using the DLP-3D printer, following similar approaches as the Microfluidics system fabrication. Results: The fabricated Bio-Ink was based onHydrogel polymer mix of sodium alginate and gelatin 15% to 0.5%, respectively. Later the 3D printing process was conducted using a higher percentage of alginate-based hydrogels because of it viscosity and the controllable crosslinking, unlike the thermal crosslinking of Gelatin. The hydrogels were colored to simulate the representation of two types of cells. The adaption of the hard scaffold, whether for the Microfluidics system or the hard-tissues, has been acquired by the DLP 3D printers with fabricated natural bioactive essential oils that contain antimicrobial activity, followed by printing in Situ three complex layers of soft-hydrogel as a cell-free Bio-Ink to simulate the real-life tissue engineering process. The final product was a proof of concept for a rapid 3D cell culturing approaches that uses an engineered hard scaffold along with soft-tissues, thus, several applications were offered as products of the current prototype, including the Organ-On-Chip as a successful integration between DLP and 3D bioprinter. Conclusion: Multiple designs for the organ-on-a-chip (multi-OoC) devices have been acquired in our study with main focus on the low cost fabrication of such technology and the potential to revolutionize human health research and development. We describe circumstances in which multi-organ models are useful after briefly examining the requirement for full multi-organ models with a systemic component. Following that, we took a look at the current multi-OoC platforms, such as integrated body-on-a-chip devices and modular techniques that use linked organ-specific modules.

Keywords: 3d bio-printer, hydrogel, multi-organ on chip, bio-inks

Procedia PDF Downloads 130

Authors: V. Schmidt, L. Janovák, N. Wiegand, B. Patczai, K. Turzó

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Large bone defects are not able to heal spontaneously. Bioactive glasses seem to be appropriate (bio)materials for bone reconstruction. Bioactive glasses are osteoconductive and osteoinductive, therefore, play a useful role in bony regeneration and repair. Because of their not optimal mechanical properties (e.g., brittleness, low bending strength, and fracture toughness), their applications are limited. Bioactive glass can be used as a coating material applied on metal surfaces. In this way -when using them as implants- the excellent mechanical properties of metals and the biocompatibility and bioactivity of glasses will be utilized. Furthermore, ion release effects of bioactive glasses regarding osteogenic and angiogenic responses have been shown. Silicate bioactive glasses (45S5 Bioglass) induce the release and exchange of soluble Si, Ca, P, and Na ions on the material surface. This will lead to special cellular responses inducing bone formation, which is favorable in the biointegration of the orthopedic prosthesis. The incorporation of other additional elements in the silicate network such as fluorine, magnesium, iron, silver, potassium, or zinc has been shown, as the local delivery of these ions is able to enhance specific cell functions. Although hip and knee prostheses present a high success rate, bacterial infections -mainly implant associated- are serious and frequent complications. Infection can also develop after implantation of hip prostheses, the elimination of which means more surgeries for the patient and additional costs for the clinic. Prosthesis-related infection is a severe complication of orthopedic surgery, which often causes prolonged illness, pain, and functional loss. While international efforts are made to reduce the risk of these infections, orthopedic surgical infections (SSIs) continue to occur in high numbers. It is currently estimated that up to 2.5% of primary hip and knee surgeries and up to 20% of revision arthroplasties are complicated by periprosthetic joint infection (PJIs). According to some authors, these numbers are underestimated, and they are also increasing. Staphylococcus aureus is the leading cause of both SSIs and PJIs, and the prevalence of methicillin-resistant S. aureus (MRSA) is on the rise, particularly in the United States. These deep infections lead to implant removal and consequently increase morbidity and mortality. The study targets this clinical problem using our experience so far with the Ag-doped polymer coatings on Titanium implants. Non-modified or modified (e.g., doped with antibacterial agents, like Ag) bioactive glasses could play a role in the prevention of infections or the therapy of infected tissues. Bioactive glasses have excellent biocompatibility, proved by in vitro cell culture studies of human osteoblast-like MG-63 cells. Ag-doped bioactive glass-scaffold has a good antibacterial ability against Escherichia coli and other bacteria. It may be concluded that these scaffolds have great potential in the prevention and therapy of implant-associated bone infection.

Keywords: antibacterial agents, bioactive glass, hip and knee prosthesis, medical implants

Procedia PDF Downloads 155

Authors: Archana Sharma, Vijayashree Nayak, Ashok Kumar

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Three-dimensional matrices were fabricated from blend of natural-natural polymers like carrageenan-gelatin and synthetic -natural polymers such as PEG- gelatin (PEG of different molecular weights (2,000 and 6,000) using two different crosslinkers; glutaraldehyde and EDC-NHS by cryogelation technique. Blends represented a feasible approach to design 3-D scaffolds with controllable mechanical, physical and biochemical properties without compromising biocompatibility and biodegradability. These matrices possessed interconnected porous structure, good mechanical strength, biodegradable nature, constant swelling kinetics, ability to withstand high temperature and visco-elastic behavior. Hemocompatibility of cryogel matrices was determined by coagulation assays and hemolytic activity assay which demonstrated that these cryogels have negligible effects on coagulation time and have excellent blood compatibility. In vitro biocompatibility (cell-matrix interaction) inferred good cell adhesion, proliferation, and secretion of ECM on matrices. These matrices provide a microenvironment for the growth, proliferation, differentiation and secretion of ECM of different cell types such as IMR-32, C2C12, Cos-7, rat bone marrow derived MSCs and human bone marrow MSCs. Hoechst 33342 and PI staining also confirmed that the cells were uniformly distributed, adhered and proliferated properly on the cryogel matrix. An ideal scaffold used for tissue engineering application should allow the cells to adhere, proliferate and maintain their functionality. Neurotransmitter analysis has been done which indicated that IMR-32 cells adhered, proliferated and secreted neurotransmitters when they interacted with these matrices which showed restoration of their functionality. The cell-matrix interaction up to molecular level was also evaluated so to check genotoxicity and protein expression profile which indicated that these cryogel matrices are non-genotoxic and maintained biofunctionality of cells growing on these matrices. All these cryogels, when implanted subcutaneously in balb/c mice, showed no adverse systemic or local toxicity effects at implantation site. There was no significant increase in inflammatory cell count has otherwise been observed after scaffold implantation. These cryogels are supermacroporous and this porous structure allows cell infiltration and proliferation of host cells. This showed the integration and presence of infiltrated cells into the cryogel implants. Histological analysis confirmed that the implanted cryogels do not have any adverse effect in spite of host immune system recognition at the site of implantation, on its surrounding tissues and other vital host organs. In vivo biocompatibility study after in vitro biocompatibility analysis has also concluded that these synthesized cryogels act as important biological substitutes, more adaptable and appropriate for transplantation. Thus, these cryogels showed their potential for soft tissue engineering applications.

Keywords: cryogelation, hemocompatibility, in vitro biocompatibility, in vivo biocompatibility, soft tissue engineering applications

Procedia PDF Downloads 194

Authors: Monica Enculescu, A. Evanghelidis, I. Enculescu

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Among the numerous methods for obtaining polymer nanofibers, the electrospinning technique distinguishes itself due to the more growing interest induced by its proved utility leading to developing and improving of the method and the appearance of novel materials. In particular, production of polymeric nanofibers in which different dopants are introduced was intensively studied in the last years because of the increased interest for the obtaining of functional electrospun nanofibers. Electrospinning is a facile method of obtaining polymer nanofibers with diameters from tens of nanometers to micrometrical sizes that are cheap, flexible, scalable, functional and biocompatible. Besides the multiple applications in medicine, polymeric nanofibers obtained by electrospinning permit manipulation of light at nanometric dimensions when doped with organic dyes or different nanoparticles. It is a simple technique that uses an electrical field to draw fine polymer nanofibers from solutions and does not require complicated devices or high temperatures. Different morphologies of the electrospun nanofibers can be obtained for the same polymeric host when different parameters of the electrospinning process are used. Consequently, we can obtain tuneable optical properties of the electrospun nanofibers (e.g. changing the wavelength of the emission peak) by varying the parameters of the fabrication method. We focus on obtaining doped polymer nanofibers with enhanced optical properties using the electrospinning technique. The aim of the paper is to produce dye-doped polymer nanofibers’ mats incorporating uniformly dispersed dyes. Transmission and fluorescence of the fibers will be evaluated by spectroscopy methods. The morphological properties of the electrospun dye-doped polymer fibers will be evaluated using scanning electron microscopy (SEM). We will tailor the luminescent properties of the material by doping the polymer (polyvinylpyrrolidone or polymethylmetacrilate) with different dyes (coumarins, rhodamines and sulforhodamines). The tailoring will be made taking into consideration the possibility of changing the luminescent properties of electrospun polymeric nanofibers that are doped with different dyes by using different parameters for the electrospinning technique (electric voltage, distance between electrodes, flow rate of the solution, etc.). Furthermore, we can evaluated the influence of the concentration of the dyes on the emissive properties of dye-doped polymer nanofibers using different concentrations. The advantages offered by the electrospinning technique when producing polymeric fibers are given by the simplicity of the method, the tunability of the morphology allowed by the possibility of controlling all the process parameters (temperature, viscosity of polymeric solution, applied voltage, distance between electrodes, etc.), and by the absence of necessity of using harsh and supplementary chemicals such as the ones used in the traditional nanofabrication techniques. Acknowledgments: The authors acknowledge the financial support received through IFA CEA Project No. C5-08/2016.

Keywords: electrospinning, luminescence, polymer nanofibers, scanning electron microscopy

Procedia PDF Downloads 180

Authors: Rangrong Yoksan, Amporn Sane, Nattaporn Khanoonkon, Chanakorn Yokesahachart, Narumol Noivoil, Khanh Minh Dang

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Polylactic acid (PLA) is the most commercially available bio-based and biodegradable plastic at present. PLA has been used in plastic related industries including single-used containers, disposable and environmentally friendly packaging owing to its renewability, compostability, biodegradability, and safety. Although PLA demonstrates reasonably good optical, physical, mechanical, and barrier properties comparable to the existing petroleum-based plastics, its brittleness and mold shrinkage as well as its price are the points to be concerned for the production of rigid and semi-rigid packaging. Blending PLA with other bio-based polymers including thermoplastic starch (TPS) is an alternative not only to achieve a complete bio-based plastic, but also to reduce the brittleness, shrinkage during molding and production cost of the PLA-based products. TPS is a material produced mainly from starch which is cheap, renewable, biodegradable, compostable, and non-toxic. It is commonly prepared by a plasticization of starch under applying heat and shear force. Although glycerol has been reported as one of the most plasticizers used for preparing TPS, its migration caused the surface stickiness of the TPS products. In some cases, mixed plasticizers or natural fibers have been applied to impede the retrogradation of starch or reduce the migration of glycerol. The introduction of fibers into TPS-based materials could reinforce the polymer matrix as well. Therefore, the objective of the present research is to study the effect of starch type (i.e. native starch and phosphate starch), plasticizer type (i.e. glycerol and xylitol with a weight ratio of glycerol to xylitol of 100:0, 75:25, 50:50, 25:75, and 0:100), and fiber content (i.e. in the range of 1-25 % wt) on properties of PLA/TPS blend and composite. PLA/TPS blends and composites were prepared using a twin-screw extruder and then converted into dumbbell-shaped specimens using an injection molding machine. The PLA/TPS blends prepared by using phosphate starch showed higher tensile strength and stiffness than the blends prepared by using the native one. In contrast, the blends from native starch exhibited higher extensibility and heat distortion temperature (HDT) than those from the modified starch. Increasing xylitol content resulted in enhanced tensile strength, stiffness, and water resistance, but decreased extensibility and HDT of the PLA/TPS blend. Tensile properties and hydrophobicity of the blend could be improved by incorporating silane treated-jute fibers.

Keywords: polylactic acid, thermoplastic starch, Jute fiber, composite, blend

Procedia PDF Downloads 397

Authors: M. Koksal, T. Ozyazici, E. Gurdal, M. Yarım, E. Demirpolat, M. B. Y. Aycan

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The discovery of new drugs in cancer chemotherapy is still a major topic because of severe side effects, selectivity problems and resistance development potential of existing drugs. In recent years, combined anticancer therapies or multi-acting drugs are clinically preferred over traditional cytotoxic treatment, with the aim of avoiding resistance and toxic side effects. Arrangement of multi-acting targets can be carried out either by combination of several drugs with different mechanisms or by usage of a single chemical compound capable of regulating several targets of a disease with multiple factors. In literature, several pyrimidine and piperazine derivatives have been involved in the structure of many compounds which have been used as chemotherapeutic agents along with wide clinical applications. The aim of this study is to combine pyrimidine and piperazine core structures to research and develop novel piperazinylpyrimidine derivatives with selective cytotoxicity over cancer cells. In this study, a group of novel 6-fluorophenyl-3-[2-(substitutedpiperazinyl)ethyl] hexahydropyrimidine-2,4-dione derivatives designed to observe the desired anticancer activity due to pyrimidine and piperazine based scaffolds. Target compounds were obtained by the reaction of appropriate piperazine derivatives and 6-(2/4-fluorophenyl)-3-(2-chloroethyl)hexahydropyrimidine-2,4-dione. The synthetic pathway of 6-(2/4-fluorophenyl)-3-(2-chloroethyl)hexahydropyrimidine-2,4-dione was started with Rodionov reaction using aldehyde, malonic acid and ammonium acetate in ethanol. Isolated β-fluorophenyl-β-amino acids were treated with 2-chloroethylisocyanate in the presence of an aqueous sodium hydroxide solution at room temperature to yield the sodium salts of the corresponding ureido acids. By addition of a mineral acid, ureido acids were precipitated. Later, these ureido acids were refluxed in thionyl chloride to give the 6-(2/4-fluorophenyl)-3-(2-chloroethyl)hexahydropyrimidine-2,4-di-one which were furthermore treated with secondary amines. Structures of purified compounds were characterized with IR, 1H-NMR, 13C-NMR, mass spectroscopies and elemental analysis. All of the compounds gave satisfactory analytical and spectroscopic data, which were in full accordance with their depicted structures. In IR spectra of the compounds, N-H group was seen at 3230-3213 cm⁻¹. C-H was seen at 3100-2820 cm⁻¹ and C=O vibrational peaks were observed approximately at 1725 and 1665 cm⁻¹ in accordance with literature. In the NMR spectra of target compounds, the methylene protons of piperazine give two separate multiplet peaks around 3.5 and 4.5 ppm representing the successful N-alkylation of the structure. The cytotoxic activity of the synthesized compounds was investigated on human bronchial epithelial (BEAS 2B), lung (A549), colon adenocarcinoma (COLO205) and breast (MCF7) cell lines, by means of sulphorhodamine B (SRB) assays in triplicate. IC₅₀ values of the screened derivatives were found in range of 11.8-78 µM. This project was supported by The Scientific and Technological Research Council of Turkey (TUBITAK, Project no: 215S157).

Keywords: cytotoxicity, hexahydropyrimidine, piperazine, sulphorhodamine B assay

Procedia PDF Downloads 130

Authors: Katerina Zaharieva, Vicente Rives, Martin Tsvetkov, Raquel Trujillano, Boris Kunev, Ivan Mitov, Maria Milanova, Zara Cherkezova-Zheleva

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The nanostructured cobalt ferrite-type materials Sample A - Co0.25Fe2.75O4, Sample B - Co0.5Fe2.5O4, and Sample C - CoFe2O4 were prepared by co-precipitation in our previous investigations. The co-precipitated Sample B and Sample C were mechanochemically activated in order to produce Sample D - Co0.5Fe2.5O4 and Sample E- CoFe2O4. The PXRD, Moessbauer and FTIR spectroscopies, specific surface area determination by the BET method, thermal analysis, element chemical analysis and temperature-programmed reduction were used to investigate the prepared nano-sized samples. The changes of the Malachite green dye concentration during reaction of the photocatalytic decolorization using nanostructured cobalt ferrite-type catalysts with different chemical composition are included. The photocatalytic results show that the increase in the degree of incorporation of cobalt ions in the magnetite host structure for co-precipitated cobalt ferrite-type samples results in an increase of the photocatalytic activity: Sample A (4 х10-3 min-1) < Sample B (5 х10-3 min-1) < Sample C (7 х10-3 min-1). Mechanochemically activated photocatalysts showed a higher activity than the co-precipitated ferrite materials: Sample D (16 х10-3 min-1) > Sample E (14 х10-3 min-1) > Sample C (7 х10-3 min-1) > Sample B (5 х10-3 min-1) > Sample A (4 х10-3 min-1). On decreasing the degree of substitution of iron ions by cobalt ones a higher sorption ability of the dye after the dark period for the co-precipitated cobalt ferrite materials was observed: Sample C (72 %) < Sample B (78 %) < Sample A (80 %). Mechanochemically treated ferrite catalysts and co-precipitated Sample B possess similar sorption capacities, Sample D (78 %) ~ Sample E (78 %) ~ Sample B (78 %). The prepared nano-sized cobalt ferrite-type materials demonstrate good photocatalytic and sorption properties. Mechanochemically activated Sample D - Co0.5Fe2.5O4 (16х10-3 min-1) and Sample E-CoFe2O4 (14х10-3 min-1) possess higher photocatalytic activity than that of the most common used UV-light catalyst Degussa P25 (12х10-3 min-1). The dependence of the photo-catalytic activity and sorption properties on the preparation method and different degree of substitution of iron ions by cobalt ions in synthesized cobalt ferrite samples is established. The mechanochemical activation leads to formation of nano-structured cobalt ferrite-type catalysts (Sample D and Sample E) with higher rate constants than those of the ferrite materials (Sample A, Sample B, and Sample C) prepared by the co-precipitation procedure. The increase in the degree of substitution of iron ions by cobalt ones leads to improved photocatalytic properties and lower sorption capacities of the co-precipitated ferrite samples. The good sorption properties between 72 and 80% of the prepared ferrite-type materials show that they could be used as potential cheap absorbents for purification of polluted waters.

Keywords: nanodimensional cobalt ferrites, photocatalyst, synthesis, mechanochemical activation

Procedia PDF Downloads 245

Authors: Paula K. Novelli, Margarida M. Barros, Luciana F. Flueri

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Utilization of agricultural by-products, wheat ban and soybean bran, as substrate for solid state fermentation (SSF) was studied, aiming the achievement of different enzymes from Aspergillus sp. with distinct biological characteristics and its application and improvement on animal nutrition. Aspergillus niger and Aspergillus oryzea were studied as they showed very high yield of phytase and protease production, respectively. Phytase activity was measure using p-nitrophenilphosphate as substrate and a standard curve of p-nitrophenol, as the enzymatic activity unit was the quantity of enzyme necessary to release one μmol of p-nitrophenol. Protease activity was measure using azocasein as substrate. Activity for phytase and protease substantially increased when the different biochemical characteristics were considered in the study. Optimum pH and stability of the phytase produced by A. niger with wheat bran as substrate was between 4.0 - 5.0 and optimum temperature of activity was 37oC. Phytase fermented in soybean bran showed constant values at all pHs studied, for optimal and stability, but low production. Phytase with both substrates showed stable activity for temperatures higher than 80oC. Protease from A. niger showed very distinct behavior of optimum pH, acid for wheat bran and basic for soybean bran, respectively and optimal values of temperature and stability at 50oC. Phytase produced by A. oryzae in wheat bran had optimum pH and temperature of 9 and 37oC, respectively, but it was very unstable. On the other hand, proteases were stable at high temperatures, all pH’s studied and showed very high yield when fermented in wheat bran, however when it was fermented in soybean bran the production was very low. Subsequently the upscale production of phytase from A. niger and proteases from A. oryzae were applied as an enzyme additive in fish fed for digestibility studies. Phytases and proteases were produced with stable enzyme activity of 7,000 U.g-1 and 2,500 U.g-1, respectively. When those enzymes were applied in a plant protein based fish diet for digestibility studies, they increased protein, mineral, energy and lipids availability, showing that these new enzymes can improve animal production and performance. In conclusion, the substrate, as well as, the microorganism species can affect the biochemical character of the enzyme produced. Moreover, the production of these enzymes by SSF can be up to 90% cheaper than commercial ones produced with the same fungi species but submerged fermentation. Add to that these cheap enzymes can be easily applied as animal diet additives to improve production and performance.

Keywords: agricultural by-products, animal nutrition, enzymes production, solid state fermentation

Procedia PDF Downloads 300

Authors: Elena I. Oprita, Oana Craciunescu, Rodica Tatia, Teodora Ciucan, Reka Barabas, Orsolya Raduly, Anca Oancea

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Healing of osteochondral defects requires repair of the damaged articular cartilage, the underlying subchondral bone and the interface between these tissues (the functional calcified layer). For this purpose, developing a single monophasic scaffold that can regenerate two specific lineages (cartilage and bone) becomes a challenge. The aim of this work was to develop variants of biodegradable cross-linked composite hydrogel based on natural polypeptides (gelatin), polysaccharides components (chondroitin-4-sulphate and hyaluronic acid), in a ratio of 2:0.08:0.02 (w/w/w) and mixed with Si-hydroxyapatite (Si-Hap), in two ratios of 1:1 and 2:1 (w/w). Si-Hap was synthesized and characterized as a better alternative to conventional Hap. Subsequently, both composite hydrogel variants were cross-linked with (N, N-(3-dimethylaminopropyl)-N-ethyl carbodiimide (EDC) and enriched with a small bioactive molecule (icariin). The small molecule icariin (Ica) (C33H40O15) is the main active constituent (flavonoid) of Herba epimedium used in traditional Chinese medicine to cure bone- and cartilage-related disorders. Ica enhances osteogenic and chondrogenic differentiation of bone marrow mesenchymal stem cells (BMSCs), facilitates matrix calcification and increases the specific extracellular matrix (ECM) components synthesis by chondrocytes. Afterward, the composite hydrogels were characterized for their physicochemical properties in terms of the enzymatic biodegradation in the presence of type I collagenase and trypsin, the swelling capacity and the degree of crosslinking (TNBS assay). The cumulative release of Ica and real-time concentration were quantified at predetermined periods of time, according to the standard curve of standard Ica, after hydrogels incubation in saline buffer at physiological parameters. The obtained cross-linked composite hydrogels enriched with small-molecule Ica were also characterized for morphology by scanning electron microscopy (SEM). Their cytocompatibility was evaluated according to EN ISO 10993-5:2009 standard for medical device testing. Thus, analyses regarding cell viability (Live/Dead assay), cell proliferation (Neutral Red assay) and cell adhesion to composite hydrogels (SEM) were performed using NCTC clone L929 cell line. The final results showed that both cross-linked composite hydrogel variants enriched with Ica presented optimal physicochemical, structural and biological properties to be used as a natural scaffold able to repair osteochondral defects. The data did not reveal any toxicity of composite hydrogels in NCTC stabilized cell lines within the tested range of concentrations. Moreover, cells were capable of spreading and proliferating on both composite hydrogel surfaces. In conclusion, the designed biodegradable cross-linked composites enriched with Si and Ica are recommended for further testing as natural temporary scaffolds, which can allow cell migration and synthesis of new extracellular matrix within osteochondral defects.

Keywords: composites, gelatin, osteochondral defect, small molecule

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Authors: Alena Semeradtova, Marcel Stofik, Lucie Mareckova, Petr Maly, Ondrej Stanek, Jan Maly

Abstract:

Recently, novel strategies based on so-called molecular evolution were shown to be effective for the production of various peptide ligand libraries with high affinities to molecular targets of interest comparable or even better than monoclonal antibodies. The major advantage of these peptide scaffolds is mainly their prevailing low molecular weight and simple structure. This study describes a new high-affinity binding molecules based immunesensor using a simple optical system for human serum albumin (HSA) detection as a model molecule. We present a comparison of two variants of recombinant binders based on albumin binding domain of the protein G (ABD) performed on micropatterned glass chip. Binding domains may be tailored to any specific target of interest by molecular evolution. Micropatterened glass chips were prepared using UV-photolithography on chromium sputtered glasses. Glass surface was modified by (3-aminopropyl)trietoxysilane and biotin-PEG-acid using EDC/NHS chemistry. Two variants of high-affinity binding molecules were used to detect target molecule. Firstly, a variant is based on ABD domain fused with TolA chain. This molecule is in vivo biotinylated and each molecule contains one molecule of biotin and one ABD domain. Secondly, the variant is ABD domain based on streptavidin molecule and contains four gaps for biotin and four ABD domains. These high-affinity molecules were immobilized to the chip surface via biotin-streptavidin chemistry. To eliminate nonspecific binding 1% bovine serum albumin (BSA) or 6% fetal bovine serum (FBS) were used in every step. For both variants range of measured concentrations of fluorescently labelled HSA was 0 – 30 µg/ml. As a control, we performed a simultaneous assay without high-affinity binding molecules. Fluorescent signal was measured using inverse fluorescent microscope Olympus IX 70 with COOL LED pE 4000 as a light source, related filters, and camera Retiga 2000R as a detector. The fluorescent signal from non-modified areas was substracted from the signal of the fluorescent areas. Results were presented in graphs showing the dependence of measured grayscale value on the log-scale of HSA concentration. For the TolA variant the limit of detection (LOD) of the optical immunosensor proposed in this study is calculated to be 0,20 µg/ml for HSA detection in 1% BSA and 0,24 µg/ml in 6% FBS. In the case of streptavidin-based molecule, it was 0,04 µg/ml and 0,07 µg/ml respectively. The dynamical range of the immunosensor was possible to estimate just in the case of TolA variant and it was calculated to be 0,49 – 3,75 µg/ml and 0,73-1,88 µg/ml respectively. In the case of the streptavidin-based the variant we didn´t reach the surface saturation even with the 480 ug/ml concentration and the upper value of dynamical range was not estimated. Lower value was calculated to be 0,14 µg/ml and 0,17 µg/ml respectively. Based on the obtained results, it´s clear that both variants are useful for creating the bio-recognizing layer on immunosensors. For this particular system, it is obvious that the variant based on streptavidin molecule is more useful for biosensing on glass planar surfaces. Immunosensors based on this variant would exhibit better limit of detection and wide dynamical range.

Keywords: high affinity binding molecules, human serum albumin, optical immunosensor, protein G, UV-photolitography

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Authors: M. Markova, E. Filova, O. Kaplan, R. Matejka, L. Bacakova

Abstract:

The porcine pericardium is used as a material for cardiac and aortic valves substitutes. Current biological aortic heart valve prosthesis have a limited lifetime period because they undergo degeneration. In order to make them more biocompatible and prolong their lifetime it is necessary to reseed the decellularized prostheses with endothelial cells and with valve interstitial cells. The endothelialization of the prosthesis-surface may be supported by suitable chemical surface modification of the prosthesis. The aim of this study is to prepare bioactive fibrin layers which would both support endothelialization of porcine pericardium and enhance differentiation and maturation of the endothelial cells seeded. As a material for surface adjustments we used layers of fibrin with/without heparin and some of them with adsorbed or chemically bound FGF2, VEGF or their combination. Fibrin assemblies were prepared in 24-well cell culture plate and were seeded with HSVEC (Human Saphenous Vein Endothelial Cells) at a density of 20,000 cells per well in EGM-2 medium with 0.5% FS and without heparin, without FGF2 and without VEGF; medium was supplemented with aprotinin (200 U/mL). As a control, surface polystyrene (PS) was used. Fibrin was also used as homogeneous impregnation of the decellularized porcine pericardium throughout the scaffolds. Morphology, density, and viability of the seeded endothelial cells were observed from micrographs after staining the samples by LIVE/DEAD cytotoxicity/viability assay kit on the days 1, 3, and 7. Endothelial cells were immunocytochemically stained for proteins involved in cell adhesion, i.e. alphaV integrin, vinculin, and VE-cadherin, markers of endothelial cells differentiation and maturation, i.e. von Willebrand factor and CD31, and for extracellular matrix proteins typically produced by endothelial cells, i.e. type IV collagen and laminin. The staining intensities were subsequently quantified using a software. HSVEC cells grew on each of the prepared surfaces better than on control surface. They reached confluency. The highest cell densities were obtained on the surface of fibrin with heparin and both grow factors used together. Intensity of alphaV integrins staining was highest on samples with remained fibrin layer, i.e. on layers with lower cell densities, i.e. on fibrin without heparin. Vinculin staining was apparent, but was rather diffuse, on fibrin with both FGF2 and VEGF and on control PS. Endothelial cells on all samples were positively stained for von Willebrand factor and CD31. VE-cadherin receptors clusters were best developed on fibrin with heparin and growth factors. Significantly stronger staining of type IV collagen was observed on fibrin with heparin and both growth factors. Endothelial cells on all samples produced laminin-1. Decellularized pericardium was homogeneously filled with fibrin structures. These fibrin-modified pericardium samples will be further seeded with cells and cultured in a bioreactor. Fibrin layers with/without heparin and with adsorbed or chemically bound FGF2, VEGF or their combination are good surfaces for endothelialization of cardiovascular prostheses or porcine pericardium based heart valves. Supported by the Ministry of Health, grants No15-29153A and 15-32497A, and the Grant Agency of the Czech Republic, project No. P108/12/G108.

Keywords: aortic valves prosthesis, FGF2, heparin, HSVEC cells, VEGF

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Authors: M. Maier, I. Dedigama, J. Majasan, Y. Wu, Q. Meyer, L. Castanheira, G. Hinds, P. R. Shearing, D. J. L. Brett

Abstract:

Increasing the efficiency of electrolyser technology is commonly seen as one of the main challenges on the way to the Hydrogen Economy. There is a significant lack of understanding of the different states of operation of polymer electrolyte membrane water electrolysers (PEMWE) and how these influence the overall efficiency. This in particular means the two-phase flow through the membrane, gas diffusion layers (GDL) and flow channels. In order to increase the efficiency of PEMWE and facilitate their spread as commercial hydrogen production technology, new analytic approaches have to be found. Acoustic emission (AE) offers the possibility to analyse the processes within a PEMWE in a non-destructive, fast and cheap in-situ way. This work describes the generation and analysis of AE data coming from a PEM water electrolyser, for, to the best of our knowledge, the first time in literature. Different experiments are carried out. Each experiment is designed so that only specific physical processes occur and AE solely related to one process can be measured. Therefore, a range of experimental conditions is used to induce different flow regimes within flow channels and GDL. The resulting AE data is first separated into different events, which are defined by exceeding the noise threshold. Each acoustic event consists of a number of consequent peaks and ends when the wave diminishes under the noise threshold. For all these acoustic events the following key attributes are extracted: maximum peak amplitude, duration, number of peaks, peaks before the maximum, average intensity of a peak and time till the maximum is reached. Each event is then expressed as a vector containing the normalized values for all criteria. Principal Component Analysis is performed on the resulting data, which orders the criteria by the eigenvalues of their covariance matrix. This can be used as an easy way of determining which criteria convey the most information on the acoustic data. In the following, the data is ordered in the two- or three-dimensional space formed by the most relevant criteria axes. By finding spaces in the two- or three-dimensional space only occupied by acoustic events originating from one of the three experiments it is possible to relate physical processes to certain acoustic patterns. Due to the complex nature of the AE data modern machine learning techniques are needed to recognize these patterns in-situ. Using the AE data produced before allows to train a self-learning algorithm and develop an analytical tool to diagnose different operational states in a PEMWE. Combining this technique with the measurement of polarization curves and electrochemical impedance spectroscopy allows for in-situ optimization and recognition of suboptimal states of operation.

Keywords: acoustic emission, gas diffusion layers, in-situ diagnosis, PEM water electrolyser

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Authors: G. M. Vlasceanu, H. Iovu, E. Vasile, M. Ionita

Abstract:

Herein, the synthesis and characterization of chitosan-gelatin highly porous scaffold reinforced with graphene oxide, and hydroxyapatite (HAp), crosslinked with genipin was targeted. In tissue engineering, chitosan and gelatin are two of the most robust biopolymers with wide applicability due to intrinsic biocompatibility, biodegradability, low antigenicity properties, affordability, and ease of processing. HAp, per its exceptional activity in tuning cell-matrix interactions, is acknowledged for its capability of sustaining cellular proliferation by promoting bone-like native micro-media for cell adjustment. Genipin is regarded as a top class cross-linker, while graphene oxide (GO) is viewed as one of the most performant and versatile fillers. The composites with natural bone HAp/biopolymer ratio were obtained by cascading sonochemical treatments, followed by uncomplicated casting methods and by freeze-drying. Their structure was characterized by Fourier Transform Infrared Spectroscopy and X-ray Diffraction, while overall morphology was investigated by Scanning Electron Microscopy (SEM) and micro-Computer Tomography (µ-CT). Ensuing that, in vitro enzyme degradation was performed to detect the most promising compositions for the development of in vivo assays. Suitable GO dispersion was ascertained within the biopolymer mix as nanolayers specific signals lack in both FTIR and XRD spectra, and the specific spectral features of the polymers persisted with GO load enhancement. Overall, correlations between the GO induced material structuration, crystallinity variations, and chemical interaction of the compounds can be correlated with the physical features and bioactivity of each composite formulation. Moreover, the HAp distribution within follows an auspicious density gradient tuned for hybrid osseous/cartilage matter architectures, which were mirrored in the mice model tests. Hence, the synthesis route of a natural polymer blend/hydroxyapatite-graphene oxide composite material is anticipated to emerge as influential formulation in bone tissue engineering. Acknowledgement: This work was supported by the project 'Work-based learning systems using entrepreneurship grants for doctoral and post-doctoral students' (Sisteme de invatare bazate pe munca prin burse antreprenor pentru doctoranzi si postdoctoranzi) - SIMBA, SMIS code 124705 and by a grant of the National Authority for Scientific Research and Innovation, Operational Program Competitiveness Axis 1 - Section E, Program co-financed from European Regional Development Fund 'Investments for your future' under the project number 154/25.11.2016, P_37_221/2015. The nano-CT experiments were possible due to European Regional Development Fund through Competitiveness Operational Program 2014-2020, Priority axis 1, ID P_36_611, MySMIS code 107066, INOVABIOMED.

Keywords: biopolymer blend, ectopic osteoinduction, graphene oxide composite, hydroxyapatite

Procedia PDF Downloads 85