Search results for: nucleic acid binding

Authors: Dmitry Nosik, Nickolay Nosik, Elli Kaplina, Olga Lobach, Marina Chataeva, Lev Rasnetsov

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Background and aims: Human Immunodeficiency Virus (HIV) infection is very often associated with Herpes Simplex Virus (HSV) infection but HIV patients are treated with a cocktail of antiretroviral drugs which are toxic. The use of an antiviral drug which will be active against both viruses like ferrovir found in our previous studies is rather actual. Earlier we had shown that Fullerene poly-amino capronic acid (FPACA) was active in case of monoinfection of HIV-1 or HSV-1. The aim of the study was to analyze the efficiency of FPACA against mixed infection of HIV and HSV. Methods: The peripheral blood lymphocytes, CEM, MT-4 cells were simultaneously infected with HIV-1 and HSV-1. FPACA was added 1 hour before infection. Cells viability was detected by MTT assay, virus antigens detected by ELISA, syncytium formation detected by microscopy. The different multiplicity of HIV-1/HSV-1 ratio was used. Results: The double viral HIV-1/HSV-1 infection was more cytopathic comparing with monoinfections. In mixed infection by the HIV-1/HSV-1 concentration of HIV-1 antigens and syncytium formations increased by 1,7 to 2,3 times in different cells in comparison with the culture infected with HIV-1 alone. The concentration of HSV-1 increased by 1,5-1,7 times, respectively. Administration of FPACA (1 microg/ml) protected cells: HIV-1/HSV-1 (1:1) – 80,1%; HIV-1/HSV-1 (1:4) – 57,2%; HIV-1/HSV-1 (1:8) – 46,3 %; HIV-1/HSV-1 (1:16) – 17,0%. Virus’s antigen levels were also reduced. Syncytium formation was totally inhibited in all cases of mixed infection. Conclusion: FPACA showed antiviral activity in case of mixed viral infection induced by Human Immunodeficiency Virus and Herpes Simplex Virus. The effect of viral inhibition increased with the multiplicity of HIV-1 in the inoculum. The mechanism of FPACA action is connected with the blocking of the virus particles adsorption to the cells and it could be suggested that it can have an antiviral activity against some other viruses too. Now FPACA could be considered as a potential drug for treatment of HIV disease complicated with opportunistic herpes viral infection.

Keywords: antiviral drug, human immunodeficiency virus (hiv), herpes simplex virus (hsv), mixed viral infection

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Authors: Tim Bolle, Franziska Kreimendahl, Thomas Gries, Stefan Jockenhoevel

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The therapeutic treatment of extensive, deep wounds is limited. Autologous split-skin grafts are used as a so-called ‘gold standard’. Most common deficits are the defects at the donor site, the risk of scarring as well as the limited availability and quality of the autologous grafts. The aim of this project is a tissue engineered dermal-epidermal skin replacement to overcome the limitations of the gold standard. A key requirement for the development of such a three-dimensional implant is the formation of a functional capillary-like network inside the implant to ensure a sufficient nutrient and gas supply. Tailored three-dimensional warp knitted spacer fabrics are used to reinforce the mechanically week fibrin gel-based scaffold and further to create a directed in vitro pre-vascularization along the parallel-oriented pile yarns within a co-culture. In this study various three-dimensional warp knitted spacer fabrics were developed in a factorial design to analyze the influence of the machine parameters such as the stitch density and the pattern of the fabric on the scaffold performance and further to determine suitable parameters for a successful fibrin gel-incorporation and a physiological performance of the scaffold. The fabrics were manufactured on a Karl Mayer double-bar raschel machine DR 16 EEC/EAC. A fine machine gauge of E30 was used to ensure a high pile yarn density for sufficient nutrient, gas and waste exchange. In order to ensure a high mechanical stability of the graft, the fabrics were made of biocompatible PVDF yarns. Key parameters such as the pore size, porosity and stress/strain behavior were investigated under standardized, controlled climate conditions. The influence of the input parameters on the mechanical and morphological properties as well as the ability of fibrin gel incorporation into the spacer fabric was analyzed. Subsequently, the pile yarns of the spacer fabrics were colonized with Human Umbilical Vein Endothelial Cells (HUVEC) to analyze the ability of the fabric to further function as a guiding structure for a directed vascularization. The cells were stained with DAPI and investigated using fluorescence microscopy. The analysis revealed that the stitch density and the binding pattern have a strong influence on both the mechanical and morphological properties of the fabric. As expected, the incorporation of the fibrin gel was significantly improved with higher pore sizes and porosities, whereas the mechanical strength decreases. Furthermore, the colonization trials revealed a high cell distribution and density on the pile yarns of the spacer fabrics. For a tailored reinforcing structure, the minimum porosity and pore size needs to be evaluated which still ensures a complete incorporation of the reinforcing structure into the fibrin gel matrix. That will enable a mechanically stable dermal graft with a dense vascular network for a sufficient nutrient and oxygen supply of the cells. The results are promising for subsequent research in the field of reinforcing mechanically weak biological scaffolds and develop functional three-dimensional scaffolds with an oriented pre-vascularization.

Keywords: fibrin-gel, skin replacement, spacer fabric, pre-vascularization

Procedia PDF Downloads 257

Authors: Meltem Haktaniyan, Eda Cagli, Irem Erel Goktepe

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Polymers that change their properties in response to different stimuli (e.g. light, temperature, pH, ionic strength or magnetic field) are called ‘smart’ or ‘stimuli-responsive polymers’. These polymers have been widely used in biomedical applications such as sensors, gene delivery, drug delivery or tissue engineering. Temperature-responsive polymers have been studied extensively for controlled drug delivery applications. As regard of pseudo-peptides, poly (2-alky-2-oxazoline)s are considered as good candidates for delivery systems due to their stealth behavior and nontoxicity. In order to build responsive multilayer films for controlled drug release applications from surface, Layer by layer technique (LBL) is a powerful technique with an advantage of nanometer scale control over spatial architecture and morphology. Multilayers can be constructed on surface where non-covalent interactions including electrostatic interactions, hydrogen bonding, and charge-transfer or hydrophobic-hydrophobic interactions. In the present study, hydrogen bounded multilayer films of poly (2-alky-2-oxazoline) s with tannic acid were prepared in order to use as a platform to release Doxorubicin (DOX) from surface with pH and thermal triggers. For this purpose, poly (2-isopropyl-2-oxazoline) (PIPOX) and poly (2-ethyl-2-oxazoline) (PETOX) were synthesized via cationic ring opening polymerization (CROP) with hydroxyl end groups. Two polymeric multilayer systems ((PETOX)/(DOX)-(TA) complexes and (PIPOX)/(DOX)-(TA) complexes) were designed to investigate of controlled release of Doxorubicin (DOX) from surface with pH and thermal triggers. The drug release profiles from the multilayer thin films with alterations of pH and temperature will been examined with UV-Vis Spectroscopy and Fluorescence Spectroscopy.

Keywords: temperature responsive polymers, h-bonded multilayer films, drug release, polyoxazoline

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Authors: Mohammadjavad Sotoudeheian

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Heart failure (HF) prevalence, as a global cardiovascular problem, is increasing gradually. A variety of molecular mechanisms contribute to HF. Proteins involved in cardiac contractility regulation, such as ion channels and calcium handling proteins, are altered. Additionally, epigenetic modifications and gene expression can lead to altered cardiac function. Moreover, inflammation and oxidative stress contribute to HF. The progression of HF can be attributed to mitochondrial dysfunction that impairs energy production and increases apoptosis. Molecular mechanisms such as these contribute to the development of cardiomyocyte defects and HF and can be therapeutically targeted. The heart's contractile function is controlled by cardiomyocytes. Calpain, and its related molecules, including Bax, VEGF, and AMPK, are among the proteins involved in regulating cardiomyocyte function. Apoptosis is facilitated by Bax. Cardiomyocyte apoptosis is regulated by this protein. Furthermore, cardiomyocyte survival, contractility, wound healing, and proliferation are all regulated by VEGF, which is produced by cardiomyocytes during inflammation and cytokine stress. Cardiomyocyte proliferation and survival are also influenced by AMPK, an enzyme that plays an active role in energy metabolism. They all play key roles in apoptosis, angiogenesis, hypertrophy, and metabolism during myocardial inflammation. The role of calpains has been linked to several molecular pathways. The calpain pathway plays an important role in signal transduction and apoptosis, as well as autophagy, endocytosis, and exocytosis. Cell death and survival are regulated by these calcium-dependent cysteine proteases that cleave proteins. As a result, protein fragments can be used for various cellular functions. By cleaving adhesion and motility proteins, calcium proteins also contribute to cell migration. HF may be brought about by calpain-mediated pathways. Many physiological processes are mediated by the calpain molecular pathways. Signal transduction, cell death, and cell migration are all regulated by these molecular pathways. Calpain is activated by calcium binding to calmodulin. In the presence of calcium, calmodulin activates calpain. Calpains are stimulated by calcium, which increases matrix metalloproteinases (MMPs). In order to develop novel treatments for these diseases, we must understand how this pathway works. A variety of myocardial remodeling processes involve calpains, including remodeling of the extracellular matrix and hypertrophy of cardiomyocytes. Calpains also play a role in maintaining cardiac homeostasis through apoptosis and autophagy. The development of HF may be in part due to calpain-mediated pathways promoting cardiomyocyte death. Numerous studies have suggested the importance of the Ca2+ -dependent protease calpain in cardiac physiology and pathology. Therefore, it is important to consider this pathway to develop and test therapeutic options in humans that targets calpain in HF. Apoptosis, autophagy, endocytosis, exocytosis, signal transduction, and disease progression all involve calpain molecular pathways. Therefore, it is conceivable that calpain inhibitors might have therapeutic potential as they have been investigated in preclinical models of several conditions in which the enzyme has been implicated that might be treated with them. Ca 2+ - dependent proteases and calpains contribute to adverse ventricular remodeling and HF in multiple experimental models. In this manuscript, we will discuss the calpain molecular pathway's important roles in HF development.

Keywords: calpain, heart failure, autophagy, apoptosis, cardiomyocyte

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Authors: Christine Fräulin, Daniela Schurr, Hamed Shahidi Rad, Gerrit Waters, Günter Rinke, Roland Dittmeyer, Michael Nilles

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The reaction mechanisms of many liquid-phase reactions in organic chemistry have not yet been sufficiently clarified. Process conditions of several hundred degrees celsius and pressures to ten megapascals complicate the sampling and the determination of kinetic data. Space resolved in-situ measurements promises new insights. A non-invasive in-situ measurement technique has the advantages that no sample preparation is necessary, there is no change in sample mixture before analysis and the sampling do no lead to interventions in the flow. Thus, the goal of our research was the development of a contact-free spatially resolved measurement technique for kinetic studies of liquid phase reaction under process conditions. Therefore we used laser Raman spectroscopy combined with an optical transparent microchannel reactor. To show the performance of the system we choose the oxidation of cyclohexane as sample reaction. Cyclohexane oxidation is an economically important process. The products are intermediates for caprolactam and adipic acid, which are starting materials for polyamide 6 and 6.6 production. To maintain high selectivities of 70 to 90 %, the reaction is performed in industry at a low conversion of about six percent. As Raman spectroscopy is usually very selective but not very sensitive the detection of the small product concentration in cyclohexane oxidation is quite challenging. To meet these requirements, an optical experimental setup was optimized to determine the concentrations by laser Raman spectroscopy with respect to good detection sensitivity. With this measurement technique space resolved kinetic studies of uncatalysed and homogeneous catalyzed cyclohexane oxidation were carried out to obtain details about the reaction mechanism.

Keywords: in-situ laser raman spectroscopy, space resolved kinetic measurements, homogeneous catalysis, chemistry

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Authors: Bianca Peterson, Tomasz J. Sańko, Carlos C. Bezuidenhout, Johnnie Van Den Berg

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Busseola fusca (Fuller) (Lepidoptera: Noctuidae), the maize stem borer, is a major pest in sub-Saharan Africa. It causes economic damage to maize and sorghum crops and has evolved non-recessive resistance to genetically modified (GM) maize expressing the Cry1Ab insecticidal toxin. Since B. fusca is a non-model organism, very little genomic information is publicly available, and is limited to some cytochrome c oxidase I, cytochrome b, and microsatellite data. The biology of B. fusca is well-described, but still poorly understood. This, in combination with its larval-specific behavior, may pose problems for limiting the spread of current resistant B. fusca populations or preventing resistance evolution in other susceptible populations. As part of on-going research into resistance evolution, B. fusca larvae were collected from Bt and non-Bt maize in South Africa, followed by RNA isolation (15 specimens) and sequencing on the Illumina HiSeq 2500 platform. Quality of reads was assessed with FastQC, after which Trimmomatic was used to trim adapters and remove low quality, short reads. Trinity was used for the de novo assembly, whereas TransRate was used for assembly quality assessment. Transcript identification employed BLAST (BLASTn, BLASTp, and tBLASTx comparisons), for which two libraries (nucleotide and protein) were created from 3.27 million lepidopteran sequences. Several transcripts that have previously been implicated in Cry toxin resistance was identified for B. fusca. These included aminopeptidase N, cadherin, alkaline phosphatase, ATP-binding cassette transporter proteins, and mitogen-activated protein kinase. MEGA7 was used to align these transcripts to reference sequences from Lepidoptera to detect mutations that might potentially be contributing to Cry toxin resistance in this pest. RSEM and Bioconductor were used to perform differential gene expression analysis on groups of B. fusca larvae challenged and unchallenged with the Cry1Ab toxin. Pairwise expression comparisons of transcripts that were at least 16-fold expressed at a false-discovery corrected statistical significance (p) ≤ 0.001 were extracted and visualized in a hierarchically clustered heatmap using R. A total of 329,194 transcripts with an N50 of 1,019 bp were generated from the over 167.5 million high-quality paired-end reads. Furthermore, 110 transcripts were over 10 kbp long, of which the largest one was 29,395 bp. BLAST comparisons resulted in identification of 157,099 (47.72%) transcripts, among which only 3,718 (2.37%) were identified as Cry toxin receptors from lepidopteran insects. According to transcript expression profiles, transcripts were grouped into three subclusters according to the similarity of their expression patterns. Several immune-related transcripts (pathogen recognition receptors, antimicrobial peptides, and inhibitors) were up-regulated in the larvae feeding on Bt maize, indicating an enhanced immune status in response to toxin exposure. Above all, extremely up-regulated arylphorin genes suggest that enhanced epithelial healing is one of the resistance mechanisms employed by B. fusca larvae against the Cry1Ab toxin. This study is the first to provide a resource base and some insights into a potential mechanism of Cry1Ab toxin resistance in B. fusca. Transcriptomic data generated in this study allows identification of genes that can be targeted by biotechnological improvements of GM crops.

Keywords: epithelial healing, Lepidoptera, resistance, transcriptome

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Authors: Hassan Beigi Rizi, Harold Auradou, Lamine Hattali

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Various industries, including civil engineering, automotive, aerospace, and biomedical fields, are currently seeking novel and innovative high-performance lightweight materials to reduce energy consumption. Inspired by the structure of Euplectella Aspergillum Glass Sponges (EA-sponge), 2D unit cells were created and fabricated using a Fused Filament Fabrication (FFF) process with Polylactic acid (PLA) filaments. The stiffness and strength of bio-inspired EA-sponge lattices were investigated both experimentally and numerically under uniaxial tensile loading and are compared to three standard square lattices with diagonal struts (Designs B and C) and non-diagonal struts (Design D) reinforcements. The aim is to establish predictive scaling laws models and examine the deformation mechanisms involved. The results indicated that for the EA-sponge structure, the relative moduli and yield strength scaled linearly with relative density, suggesting that the deformation mechanism is stretching-dominated. The Finite element analysis (FEA), with periodic boundary conditions for volumetric homogenization, confirms these trends and goes beyond the experimental limits imposed by the FFF printing process. Therefore, the stretching-dominated behavior, investigated from 0.1 to 0.5 relative density, demonstrate that the study of EA-sponge structure can be exploited for the realization of square lattice topologies that are stiff and strong and have attractive potential for lightweight structural applications. However, the FFF process introduces an accuracy limitation, with approximately 10% error, making it challenging to print structures with a relative density below 0.2. Future work could focus on exploring the impact of different printing materials on the performance of EA-sponge structures.

Keywords: bio-inspiration, lattice structures, fused filament fabrication, scaling laws

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Authors: Karl Kingsley

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Many mechanisms are involved in the control of cellular differentiation and growth, which are often dysregulated in many cancers. Many distinct pathways are involved in these mechanisms of control, including deoxyribonuclease (DNA) methyltransferase and histone deacetylase (HDAC) activation that controls both genetic and epigenetic modifications and micro ribonucleic acid (RNA) expression. Less is known about the expression of DNA methyltransferase (DNMT) and HDAC in oral cancers and the effect on microRNA expression. The primary objective of this study was to evaluate the expression of DNMT and HDAC family members in oral cancer and the concomitant expression of cancer-associated microRNAs. Using commercially available oral cancers, including squamous cell carcinoma (SCC)-4, SCC-9, SCC-15, and SCC-25, RNA was extracted and screened for DNMT, HDAC, and microRNA expression using highly-specific primers and quantitative polymerase chain reaction (qPCR). These data revealed low or absent expression of DNMT-1, which is associated with cellular differentiation but increased expression of DNMT-3a and DNMT-3b in all SCC cell lines compared with normal non-cancerous cell controls. In addition, no expression of HDAC1 and HDAC2 expression was found among the normal, non-cancerous cells but was highly expressed in each of the SCC cell lines examined. Differential expression of oncogenic and cancer-associated microRNAs was also observed among the SCC cell lines, including miR-21, miR-133, miR-149, miR-155, miR-365, and miR-720. These findings also appeared to vary according to observed growth rates among these cells. These data may be the first to demonstrate the expression and association between HDAC and DNMT3 family members among oral cancers. In addition, the differential expression of these epigenetic modifiers may be associated with the expression of specific microRNAs in these cancers, which have not previously been observed to the best of the author's knowledge. In addition, some associations and relationships may exist between the expression of these biomarkers and the rates of growth and proliferation, which may suggest that these expression patterns might represent potentially useful biomarkers to determine tumor aggressiveness and other phenotypic behaviors among oral cancers.

Keywords: oral cancer, DNA methyltransferase, histone deacetylase, microRNA

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Authors: Saad M. Howladar, Mostafa M. Rady, Wael M. Semida

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A field experiment was conducted during the two successive seasons of 2012 and 2013 in the Experimental Farm (newly-reclaimed saline soil; EC = 7.8 dS m-1), Faculty of Agriculture, Fayoum University, Fayoum, Egypt to investigate the effect of vitamin C foliar application at the rates of 1, 2, 3 and 4 mM on the possibility of improving growth, seed and oil yields, and some chemical constituents of Helianthus annuus L. plants under the adverse conditions of the selected soil. Significant positive influences of all vitamin C treatments were observed on growth, seed and oil yields and some chemical constituents in both seasons. Compared to unsprayed plants (control), spraying plants with various rates of vitamin C significantly increased vegetative growth traits (i.e. plant height, No. of leaves plant-1, leaf area leaf-1, total leaves area plant-1, and dry weights of leaves and shoot plant-1) and seed and oil yields and their components (i.e. head diameter, seed weight head-1, 100-seed weight, seed yield feddan-1 and oil yield feddan-1). In addition, the concentrations of chlorophyll a, chlorophyll b, total chlorophylls, total carotenoids and total phenols in fresh leaves, and total carbohydrates, total soluble sugars, free proline and some nutrients (i.e. N, P, K, Fe, Mn, and Zn) in dry leaves were also increased significantly with all vitamin C applications. Vitamin C treatment at the rate of 3 mM was generated the best results. These results are important as the potential of vitamin C to alleviate the harmful effects of salt stress offer an opportunity to increase the resistance of sunflower plants to grow under saline conditions of the newly-reclaimed soils.

Keywords: sunflower, Helianthus annuus L., ascorbic acid, salinity, growth, seed yield, oil content, chemical composition

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Authors: Memoona Ramzan, Bushra Tabassum, Anwar Khan, Muhammad Tariq, Mudassar Fareed Awan, Idrees Ahmad Nasir, Zahida Qamar, Naila Shahid, Tayyab Husnain

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Biological control is a novel approach being used in crop protection nowadays. Bacteria like Bacillus and Pseudomonas are reported for this purpose and few of their products are commercially available too. Rhizosphere and phyllosphere of healthy cotton plants were used as a source to isolate bacteria capable of exhibiting properties worthy for selection as biocontrol agent. For this purpose all isolated strains were screened for the activities like phosphate solubilization, Indole acetic acid (IAA) production and biocontrol against fungi. Two strains S1HL3 and S1HL4 showed phosphate solubilization and IAA production simultaneously while two other JS2HR4 and JS3HR2 were good inhibitors of fungal pathogens. Through biochemical and molecular characterization these bacteria were identified as P. aeruginosa, Burkholderia and Bacillus respectively. In green house trials of these isolates against Cotton leaf curl virus (CLCuV), seven treatments including individual bacterial isolate and consortia were included. Treated plants were healthy as compared to control plants in which upto 74% CLCuV symptomatic plants exist. Maximum inhibition of CLCuV was observed in T7 treated plants where viral load was only 0.4% as compared to control where viral load was upto 74%. This treatment consortium included Bacillus and Pseudomonas isolates; S1HL3, S1HL4, JS2HR4 and JS3HR2. Principal Component Biplot depicted highly significant correlation between percentage viral load and the disease incidence.

Keywords: cotton leaf curl virus, biological control, bacillus, pseudomonas

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Authors: Akhtar Aman, Abida Raza, Shumaila Bashir, Javaid Irfan, Andreas G. Schätzlein, Ijeoma F Uchegbeu

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Development of efficient delivery system for hydrophobic drugs remains a major concern in chemotherapy. The objective of the current study was to develop polymeric drug-delivery system for etoposide from amphiphilic derivatives of glycol chitosan, capable to improve the pharmacokinetics and to reduce the adverse effects of etoposide due to various organic solvents used in commercial formulations for solubilisation of etoposide. As a promising carrier, amphiphilic derivatives of glycol chitosan were synthesized by chemical grafting of palmitic acid N-hydroxy succinimide and quaternisation to glycol chitosan backbone. To this end a 7.9 kDa glycol chitosan was modified by palmitoylation and quaternisation into 13 kDa. Nano sized micelles prepared from this amphiphilic polymer had the capability to encapsulate up to 3 mg/ml etoposide. The pharmacokinetic results indicated that GCPQ based etoposide formulation transformed the biodistribution pattern. AUC 0.5-24 hr showed statistically significant difference in ETP-GCPQ vs. commercial preparation in liver (25 vs 70, p<0.001), spleen (27 vs. 36, P<0.05), lungs (42 vs. 136, p<0.001), kidneys (25 vs. 30, p<0.05) and brain (19 vs. 9,p<0.001). Using the hydrophobic fluorescent dye Nile red, we showed that micelles efficiently delivered their payload to MCF7 and A2780 cancer cells in-vitro and to A431 xenograft tumor in-vivo, suggesting these systems could deliver hydrophobic anti- cancer drugs such as etoposide to tumors. The pharmacokinetic results indicated that the GCPQ micelles transformed the biodistribution pattern and increased etoposide concentration in the brain significantly compared to free drug after intravenous administration. GCPQ based formulations not only reduced side effects associated with current available formulations but also increased their transport through the biological barriers, thus making it a good delivery system.

Keywords: glycol chitosan, Nile red, micelles, etoposide, A431 xenografts

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Authors: Cristian Moisa, Andreea Lupitu, Adriana Csakvari, Dana G. Radu, Leonard Marian Olariu, Georgeta Pop, Dorina Chambre, Lucian Copolovici, Dana Copolovici

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Green synthesis of nanoparticles (NPs) represents a promising, accessible, eco-friendly, and safe process with significant applications in biotechnology, pharmaceutical sciences, and farming. The aim of our study was to obtain silver nanoparticles, using plant wastes extracts resulted in the essential oils extraction process: Thymus vulgaris L., Origanum vulgare L., Lavandula angustifolia L., and in hemp processing for seed and fibre, Cannabis sativa. Firstly, we obtained aqueous extracts of thyme, oregano, lavender, and hemp (two monoicous and one dioicous varieties), all harvested in western part of Romania. Then, we determined the chemical composition of the extracts by liquid-chromatography coupled with diode array and mass spectrometer detectors. The compounds identified in the extracts were in agreement with earlier published data, and the determination of the antioxidant activity of the obtained extracts by DPPH (2,2-diphenyl-1-picrylhydrazyl) and ABTS (2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)) assays confirmed their antioxidant activity due to their total polyphenolic content evaluated by Folin-Ciocalteu assay. Then, the silver nanoparticles (AgNPs) were successfully biosynthesised, as was demonstrated by UV-VIS, FT-IR spectroscopies, and SEM, by reacting AgNO₃ solution and plant extracts. AgNPs were spherical in shape, with less than 30 nm in diameter, and had a good bactericidal activity against Gram-positive (Staphylococcus aureus) and Gram-negative bacteria (Escherichia coli, Klebsiella pneumoniae, Pseudomonas fluorescens).

Keywords: plant wastes extracts, chemical composition, high performance liquid chromatography mass spectrometer, HPLC-MS, scanning electron microscopy, SEM, silver nanoparticles

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Authors: Shanna Swart, Caryn Fenner, Athanasios Kotsiopoulos, Susan Harrison

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Linear alkanes are produced as by-products from the increasing use of gas-to-liquid fuel technologies for synthetic fuel production and offer great potential for value addition. Their current use as low-value fuels and solvents do not maximize this potential. Therefore, attention has been drawn towards direct activation of these aliphatic alkanes to more useful products such as alcohols, aldehydes, carboxylic acids and derivatives. Cytochrome P450 monooxygenases (P450s) can be used for activation of these aliphatic alkanes using whole-cells or cell-free systems. Some limitations of whole-cell systems include reduced mass transfer, stability and possible side reactions. Since the P450 systems are little studied as cell-free systems, they form the focus of this study. Challenges of a cell-free system include co-factor regeneration, substrate availability and enzyme stability. Enzyme immobilization offers a positive outlook on this dilemma, as it may enhance stability of the enzyme. In the present study, 2 different P450s (CYP153A6 and CYP102A1) as well as the relevant accessory enzymes required for electron transfer (ferredoxin and ferredoxin reductase) and co-factor regeneration (glucose dehydrogenase) have been expressed in E. coli and purified by metal affinity chromatography. Glucose dehydrogenase (GDH), was used as a model enzyme to assess the potential of various enzyme immobilization strategies including; surface attachment on MagReSyn® microspheres with various functionalities and on electrospun nanofibers, using self-assembly based methods forming Cross Linked Enzymes (CLE), Cross Linked Enzyme Aggregates (CLEAs) and spherezymes as well as in a sol gel. The nanofibers were synthesized by electrospinning, which required the building of an electrospinning machine. The nanofiber morphology has been analyzed by SEM and binding will be further verified by FT-IR. Covalent attachment based methods showed limitations where only ferredoxin reductase and GDH retained activity after immobilization which were largely attributed to insufficient electron transfer and inactivation caused by the crosslinkers (60% and 90% relative activity loss for the free enzyme when using 0.5% glutaraldehyde and glutaraldehyde/ethylenediamine (1:1 v/v), respectively). So far, initial experiments with GDH have shown the most potential when immobilized via their His-tag onto the surface of MagReSyn® microspheres functionalized with Ni-NTA. It was found that Crude GDH could be simultaneously purified and immobilized with sufficient activity retention. Immobilized pure and crude GDH could be recycled 9 and 10 times, respectively, with approximately 10% activity remaining. The immobilized GDH was also more stable than the free enzyme after storage for 14 days at 4˚C. This immobilization strategy will also be applied to the P450s and optimized with regards to enzyme loading and immobilization time, as well as characterized and compared with the free enzymes. It is anticipated that the proposed immobilization set-up will offer enhanced enzyme stability (as well as reusability and easy recovery), minimal mass transfer limitation, with continuous co-factor regeneration and minimal enzyme leaching. All of which provide a positive outlook on this robust multi-enzyme system for efficient activation of linear alkanes as well as the potential for immobilization of various multiple enzymes, including multimeric enzymes for different bio-catalytic applications beyond alkane activation.

Keywords: alkane activation, cytochrome P450 monooxygenase, enzyme catalysis, enzyme immobilization

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Authors: Vladimir A. Basiuk, Laura J. Flores-Sanchez, Victor Meza-Laguna, Jose O. Flores-Flores, Lauro Bucio-Galindo, Elena V. Basiuk

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Noncovalent nanohybrid materials combining carbon nanotubes (CNTs) with phthalocyanines (Pcs) is a subject of increasing research effort, with a particular emphasis on the design of new heterogeneous catalysts, efficient organic photovoltaic cells, lithium batteries, gas sensors, field effect transistors, among other possible applications. The possibility of using unsubstituted Pcs for CNT functionalization is very attractive due to their very moderate cost and easy commercial availability. However, unfortunately, the deposition of unsubstituted Pcs onto nanotube sidewalls through the traditional liquid-phase protocols turns to be very problematic due to extremely poor solubility of Pcs. On the other hand, unsubstituted free-base H₂Pc phthalocyanine ligand, as well as many of its transition metal complexes, exhibit very high thermal stability and considerable volatility under reduced pressure, which opens the possibility for their physical vapor deposition onto solid surfaces, including nanotube sidewalls. In the present work, we show the possibility of simple, fast and efficient noncovalent functionalization of single-walled carbon nanotubes (SWNTs) with a series of 3d metal(II) phthalocyanines Me(II)Pc, where Me= Co, Ni, Cu, and Zn. The functionalization can be performed in a temperature range of 400-500 °C under moderate vacuum and requires about 2-3 h only. The functionalized materials obtained were characterized by means of Fourier-transform infrared (FTIR), Raman, UV-visible and energy-dispersive X-ray spectroscopy (EDS), scanning and transmission electron microscopy (SEM and TEM, respectively) and thermogravimetric analysis (TGA). TGA suggested that Me(II)Pc weight content is 30%, 17% and 35% for NiPc, CuPc, and ZnPc, respectively (CoPc exhibited anomalous thermal decomposition behavior). The above values are consistent with those estimated from EDS spectra, namely, of 24-39%, 27-36% and 27-44% for CoPc, CuPc, and ZnPc, respectively. A strong increase in intensity of D band in the Raman spectra of SWNT‒Me(II)Pc hybrids, as compared to that of pristine nanotubes, implies very strong interactions between Pc molecules and SWNT sidewalls. Very high absolute values of binding energies of 32.46-37.12 kcal/mol and the highest occupied and lowest unoccupied molecular orbital (HOMO and LUMO, respectively) distribution patterns, calculated with density functional theory by using Perdew-Burke-Ernzerhof general gradient approximation correlation functional in combination with the Grimme’s empirical dispersion correction (PBE-D) and the double numerical basis set (DNP), also suggested that the interactions between Me(II) phthalocyanines and nanotube sidewalls are very strong. The authors thank the National Autonomous University of Mexico (grant DGAPA-IN200516) and the National Council of Science and Technology of Mexico (CONACYT, grant 250655) for financial support. The authors are also grateful to Dr. Natalia Alzate-Carvajal (CCADET of UNAM), Eréndira Martínez (IF of UNAM) and Iván Puente-Lee (Faculty of Chemistry of UNAM) for technical assistance with FTIR, TGA measurements, and TEM imaging, respectively.

Keywords: carbon nanotubes, functionalization, gas-phase, metal(II) phthalocyanines

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Authors: Hichem Sebai, Mohamed Amine Jabri, Kais Rtibi, Haifa Tounsi, Lamjed Marzouki

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Artemisia campestris has been widely used in Tunisian traditional medicine for its health beneficial effects. However, the present study aims at evaluating the antiulcer effects of Artemisia campestris aqueous extract (ACAE) as well as the mechanism of action involved in such gastroprotection. In this respect, male Wistar rats were divided into seven groups: control, aspirin (ASPR), ASPR + various doses of ACAE (100, 200 and 400 mg/kg, b.w.), ASPR+ famotidine and ASPR+ caffeic acid. Animals were pre-treated with ACAE extract during 10 days. We firstly showed that aspirin administration was accompanied by an oxidative stress status assessed by an increase of malondialdehyde (MDA) level, a decrease of sulfhydryl -(SH) groups content and depletion of antioxidant enzyme activities such as superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx). Pre-treatment with ACAE protected against aspirin-induced gastric oxidative stress. More importantly, aspirin administration increased plasma and tissue hydrogen peroxide (H₂O₂), free iron and calcium levels while the ACAE pre-treatment reversed all aspirin-induced intracellular mediators disturbance. The results of the present study clearly indicated that AEAC gastroprotection might be related, at least in part, to its antioxidant properties as well as to various gastric mucosal defense mechanisms, including the protection of gastric sulfhydryls and an opposite effect on some intracellular mediators such as free iron, hydrogen peroxide, and calcium. However, our data confirm the use of Artemisia campestris extracts in the Tunisian traditional folk medicine for the treatment of gastrointestinal diseases.

Keywords: gastric ulcer, Artemisia campestris, oxidative stress, sulfhydryl groups, Fenton reaction, rat

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Authors: Alla Mariseva, Ilze Beitane

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Daily intake of products rich in antioxidants that scavenge free radicals in cell membranes is an effective way to combat oxidative stress. Last year there was noticed higher interest towards the identification and utilization of plants rich in antioxidant compounds as they may behave as preventive medicine. Amaranth seeds due to polyphenols, anthocyanins, flavonoids, and tocopherols are characterized by high antioxidant activity. The study aimed to evaluate the total phenolic content and radical scavenging activity of amaranth seeds cultivated in 2020 in two farms in Latvia. One sample of amaranth seeds came from an organic farm, the other – from a conventional farm. The total phenol content of amaranth seed extracts was measured with the Folin-Ciocalte spectrophotometric method. The total phenols were expressed as gallic acid equivalents (GAE) per 100 g dry weight (DW) of the samples. The antioxidant activity of amaranth seed extracts was calculated based on scavenging activities of the stable 2.2-diphenyl-1-picrylhydrazyl (DPPH˙) radical, the radical scavenging capacity (ABTS) was demonstrated as Trolox mM equivalents (TE) per 100 g-1 dry weight. Three parallel measurements were performed on all samples. There were significant differences between organic and conventional amaranth seeds in terms of total phenolic content and antioxidant activity. Organic amaranth seeds showed higher total phenolic content compared to conventional amaranth seeds, 65.4±6.0 mg GAE 100 g⁻¹ DW and 43.4±7.8 mg GAE 100 g⁻¹ DW respectively. Organic amaranth seeds were also characterized by higher DPPH radical scavenging activity (7.9±0.4 mM TE 100 g⁻¹ of dry matter) and ABTS radical scavenging capacity (13.2±1.5 mM TE 100 g⁻¹ of dry matter). The results obtained on total phenolic content and antioxidant activity of amaranth seeds grown in Latvia confirmed that the samples have a high biological value; therefore, it would be necessary to promote their consumption by including them in various food products, including vegan products, increasing their nutritional value.

Keywords: ABTS, amaranth seeds, antioxidant activity, DPPH, total phenolic content

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Authors: Olia Kanevskaia

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Standardization has been in the limelight of numerous academic studies. Typically described as ‘any set of technical specifications that either provides or is intended to provide a common design for a product or process’, standards do not only set quality benchmarks for products and services, but also spur competition and innovation, resulting in advantages for manufacturers and consumers. Their contribution to globalization and technology advancement is especially crucial in the Information and Communication Technology (ICT) and telecommunications sector, which is also characterized by a weaker state-regulation and expert-based rule-making. Most of the standards developed in that area are interoperability standards, which allow technological devices to establish ‘invisible communications’ and to ensure their compatibility and proper functioning. This type of standard supports a large share of our daily activities, ranging from traffic coordination by traffic lights to the connection to Wi-Fi networks, transmission of data via Bluetooth or USB and building the network architecture for the Internet of Things (IoT). A large share of ICT standards is developed in the specialized voluntary platforms, commonly referred to as Standards Development Organizations (SDOs), which gather experts from various industry sectors, private enterprises, governmental agencies and academia. The institutional architecture of these bodies can vary from semi-public bodies, such as European Telecommunications Standards Institute (ETSI), to industry-driven consortia, such as the Internet Engineering Task Force (IETF). The past decades witnessed a significant shift of standard setting to those institutions: while operating independently from the states regulation, they offer a rather informal setting, which enables fast-paced standardization and places technical supremacy and flexibility of standards above other considerations. Although technical norms and specifications developed by such nongovernmental platforms are not binding, they appear to create significant regulatory impact. In the United States (US), private voluntary standards can be used by regulators to achieve their policy objectives; in the European Union (EU), compliance with harmonized standards developed by voluntary European Standards Organizations (ESOs) can grant a product a free-movement pass. Moreover, standards can de facto manage the functioning of the market when other regulative alternatives are not available. Hence, by establishing (potentially) mandatory norms, SDOs assume regulatory functions commonly exercised by States and shape their own legal order. The purpose of this paper is threefold: First, it attempts to shed some light on SDOs’ institutional architecture, focusing on private, industry-driven platforms and comparing their regulatory frameworks with those of formal organizations. Drawing upon the relevant scholarship, the paper then discusses the extent to which the formulation of technological standards within SDOs constitutes a private legal order, operating in the shadow of governmental regulation. Ultimately, this contribution seeks to advise whether a state-intervention in industry-driven standard setting is desirable, and whether the increasing regulatory importance of SDOs should be addressed in legislation on standardization.

Keywords: private order, standardization, standard-setting organizations, transnational law

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Authors: Krasimira Georgieva, Yordan Denev

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Many of thermosetting resins have application only in filled state, reinforced with different mineral fillers. The co-filling of polymers with mineral filler and gases creates a possibility for production of polymer composites materials with low density. This processing leads to forming of new materials – gas-filled plastics (polymer foams). The properties of these materials are determined mainly by the shape and size of internal structural elements (pores). The interactions on the phase boundaries have influence on the materials properties too. In the present work, the gas-filled urea-formaldehyde resins were reinforced by waste phosphogypsum. The waste phosphogypsum (CaSO₄.2H₂O) is a solid by-product in wet phosphoric acid production processes. The values of the interactions polymer-filler were increased by using two modifying agents: polyvinyl acetate for polymer matrix and sodium metasilicate for filler. Technological methods for gas-filling and recipes of urea-formaldehyde based materials with apparent density 20-120 kg/m³ were developed. The heat conductivity of the samples is between 0.024 and 0.029 W/m^oK. Tensile analyses were carried out at 10 and 50% deformation and show values 0.01-0.14 MPa and 0.01-0.09 MPa, respectively. The apparent density of obtained materials is between 20 and 92 kg/m³. The changes in the tensile properties and density of these materials according to sodium metasilicate content were studied too. The mechanism of phosphogypsum adsorption modification was studied using methods of FT-IR spectroscopy. The structure of the gas-filled urea-formaldehyde resins was described by results of electron scanning microscopy at three different magnification ratios – x50, x150 and x 500. The aim of present work is to study the possibility of the usage of phosphogypsum as mineral filler for urea-formaldehyde resins and development of a technology for the production of gas-filled reinforced polymer composite materials. The structure and the properties of obtained composite materials are suitable for thermal and sound insulation applications.

Keywords: urea formaldehyde resins, gas-filled thermostes, phosphogypsum, mechanical properties

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Authors: Hamza Hadj Abdallah, Brahim Belabdi

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Respiratory disease is a dominant pathology in cattle. It causes mortality and especially morbidity and irreversible damage. Although the dairy herd is affected, it is essentially the lactating herd and especially young cattle either nursing or fattening that undergo the greatest economic impact. The objective of this study is to establish a microbiological diagnosis of bovine respiratory inffections from lung presented with gross lesions at the slaughter of Batna. A total of 124 samples (pharyngeal and nasal swabs and lung fragments) from 31 seven months old calves, with lung lesions was collected to determine possible correlations between etiologic agents and lesion types. The hépatisation injury (or consolidation) was the major lesion (45.17%) preferentially localized in the right apical lobe. A diverse microbial flora (15 genera and 291 strains was isolated. The bacteria most frequently isolated are the Enterobacteriaceae (49.45%), Staphylococci (25.1%) followed by non Enterobacteriaceae bacilli represented by Pseudomonas (5.83%) and finally, Streptococcus (13.38 %). The pneumotropic bacteria (Pasteurellaaerogenes and Pasteurellapneumotropica) were isolated at a rate of 0.68%. The study of the sensitivity of some germs to antibiotics showed a sensitivity of 100% for ceftazidime. A very high sensitivity was also observed for kanamycin, Ciprofloxacin, Imepinem, Cefepime, Tobramycin and Gentamycin (between 90% and 97%). Strains of E. coli showed a sensitivity of 100% for Imepinem, while only 55.9% of the strains were sensitive to Ampicillin. The isolated Pasteurella exhibited excellent sensitivity (100%) for the antimicrobials used with the exception of Colistin and Ticarcillin-Clavulanic acid association which showed a sensitivity of 50%.This survey has demonstrated the strong spread of atypical pneumonia in cattle population (bulls) at the slaughterhouse of Batna justifying stunting and losses in cattle farms in the region.Thus, it was considered urgent to establish a profile of sensitivity of different germs to antibiotics isolated to limit this increasingly dreadful infection.

Keywords: Pasteurella, enterobacteria, bacteriology, pneumonia

Procedia PDF Downloads 221

Authors: R. S. Komartin, B. Balanuca, R. Stan

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the last decades, studies about the use of polymeric materials of plant origin, considering environmental concerns, have captured the interest of researchers because these represent an alternative to petroleum-derived materials. Vegetable oils are one of the preferred alternatives for petroleum-based raw materials having long aliphatic chains similar to hydrocarbons which means that can be processed using conventional chemistry. Epoxidized vegetable oils (EVO) are among the most interesting products derived from oil both for their high reactivity (epoxy group) and for the potential to react with compounds from various classes. As in the case of epoxy resins starting from petrochemical raw materials, those obtained from EVO can be crosslinked with different agents to build polymeric networks and can also be reinforced with various additives to improve their thermal and mechanical performances. Among the multitude of known EVO, the most common in industrial practice are epoxidized linseed oils (ELO) and epoxidized soybean oils (ESO), the first with an iodine index over 180, the second having a lower iodine index but being cheaper. On the other hand, lignin (Ln) is the second natural organic material as a spread, whose use has long been hampered because of the high costs associated with its isolation and purification. In this context, our goal was to obtain new composite materials with satisfactory intermediate properties in terms of stiffness and elasticity using the characteristics of ELO and Ln and choosing the proper curing procedure. In the present study linseed oil (LO) epoxidation was performed using peracetic acid generated in situ. The obtained bio-based epoxy resin derived from linseed oil was used further to produce the new composites byloading Ln in various mass ratios. The resulted ELO-Ln blends were subjected to a dual-curing protocol, namely photochemical and thermal. The new ELO-Ln composites were investigated by FTIR spectrometry, thermal stability, water affinity, and morphology. The positive effect of lignin regarding the thermal stability of the composites could be proved. The results highlight again the still largely unexplored potential of lignin in industrial applications.

Keywords: composite materials, dual curing, epoxidized linseed oil, lignin

Procedia PDF Downloads 158

Authors: Xiaodi Su, Xin Ting Zheng, Laura Sutarlie, Nur Asinah binte Mohamed Salleh, Yong Yu

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Wound healing is a dynamic process with multiple phases. Rapid profiling and quantitative characterization of inflammation and infection remain challenging. We have developed paper-like battery-free multiplexed sensors for holistic wound assessment via quantitative detection of multiple inflammation and infection markers. In one of the designs, the sensor patch consists of a wax-printed paper panel with five colorimetric sensor channels arranged in a pattern resembling a five-petaled flower (denoted as a ‘Petal’ sensor). The five sensors are for temperature, pH, trimethylamine, uric acid, and moisture. The sensor patch is sandwiched between a top transparent silicone layer and a bottom adhesive wound contact layer. In the second design, a palm-like-shaped paper strip is fabricated by a paper-cutter printer (denoted as ‘Palm’ sensor). This sensor strip carries five sensor regions connected by a stem sampling entrance that enables rapid colorimetric detection of multiple bacteria metabolites (aldehyde, lactate, moisture, trimethylamine, tryptophan) from wound exudate. For both the “\’ Petal’ and ‘Palm’ sensors, color images can be captured by a mobile phone. According to the color changes, one can quantify the concentration of the biomarkers and then determine wound healing status and identify/quantify bacterial species in infected wounds. The ‘Petal’ and ‘Palm’ sensors are validated with in-situ animal and ex-situ skin wound models, respectively. These sensors have the potential for integration with wound dressing to allow early warning of adverse events without frequent removal of the plasters. Such in-situ and early detection of non-healing condition can trigger immediate clinical intervention to facilitate wound care management.

Keywords: wound infection, colorimetric sensor, paper fluidic sensor, wound care

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Authors: Liliane Samara Ferreira Leite, Francys Kley Vieira Moreira, Luiz Henrique Capparelli Mattoso

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The increasing environmental concern regarding the plastic pollution worldwide has stimulated the development of low-cost biodegradable materials. Proteins are renewable feedstocks that could be used to produce biodegradable plastics. Gelatin, for example, is a cheap film-forming protein extracted from animal skin and connective tissues of Brazilian Livestock residues; thus it has a good potential in low-cost biodegradable plastic production. However, gelatin plastics are limited in terms of mechanical and barrier properties. Cellulose nanocrystals (CNC) are efficient nanofillers that have been used to extend physical properties of polymers. This work was aimed at evaluating the reinforcing efficiency of CNC on gelatin films. Specifically, we have employed the continuous casting as the processing method for obtaining the gelatin/CNC bionanocomposites. This required a first rheological study for assessing the effect of gelatin-CNC and CNC-CNC interactions on the colloidal state of the aqueous bionanocomposite formulations. CNC were isolated from eucalyptus pulp by sulfuric acid hydrolysis (65 wt%) at 55 °C for 30 min. Gelatin was solubilized in ultra-pure water at 85°C for 20 min and then mixed with glycerol at 20 wt.% and CNC at 0.5 wt%, 1.0 wt% and 2.5 wt%. Rotational measurements were performed to determine linear viscosity (η) of bionanocomposite solutions, which increased with increasing CNC content. At 2.5 wt% CNC, η increased by 118% regarding the neat gelatin solution, which was ascribed to percolation CNC network formation. Storage modulus (G’) and loss modulus (G″) further determined by oscillatory tests revealed that a gel-like behavior was dominant in the bionanocomposite solutions (G’ > G’’) over a broad range of temperature (20 – 85 °C), particularly at 2.5 wt% CNC. These results confirm effective interactions in the aqueous gelatin-CNC bionanocomposites that could substantially increase the physical properties of the gelatin plastics. Tensile tests are underway to confirm this hypothesis. The authors would like to thank the Fapesp (process n 2016/03080-3) for support.

Keywords: bionanocomposites, cellulose nanocrystals, gelatin, viscoelastic characterization

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Authors: Hamideh Hamedi, Nima Rezaei, Sohrab Zendehboudi

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Separating emulsified oil contaminations from waste- or produced water is of interest to various industries. Magnetic particles (MPs) application for separating dispersed and emulsified oil from wastewater is becoming more popular. Stabilization of MPs is required through developing a coating layer on their surfaces to prevent their agglomeration and enhance their dispersibility. In this research, we study the effects of coating material, size, and concentration of iron oxide MPs on oil separation efficiency, using oil adsorption capacity measurements. We functionalize both micro-and nanoparticles of Fe3O4 using sodium dodecyl sulfate (SDS) as an anionic surfactant, cetyltrimethylammonium bromide (CTAB) as a cationic surfactant, and stearic acid (SA). The chemical structures and morphologies of these particles are characterized using Scanning Electron Microscopy (SEM), Transmission Electron Microscopy (TEM), and Energy Dispersive X-ray (EDX). The oil-water separation results indicate that a low dosage of the coated magnetic nanoparticle with CTAB (0.5 g/L MNP-CTAB) results the highest oil adsorption capacity (nearly 100%) for 1000 ppm dodecane-in-water emulsion, containing ultra-small droplets (250–300 nm). While separation efficiency of the same dosage of bare MNPs is around 57.5%. Demulsification results of magnetic microparticles (MMPs) also reveal that the functionalizing particles with CTAB increase oil removal efficiency from 86.3% for bare MMP to 92% for MMP-CTAB. Comparing the results of different coating materials implies that the major interaction reaction is an electrostatic attraction between negatively charged oil droplets and positively charged MNP-CTAB and MMP-CTAB. Furthermore, the synthesized nanoparticles could be recycled and reused; after ten cycles the oil adsorption capacity slightly decreases to near 95%. In conclusion, functionalized magnetic particles with high oil separation efficiency could be used effectively in treatment of oily wastewater. Finally, optimization of the adsorption process is required by considering the effective system variables, and fluid properties.

Keywords: oily wastewater treatment, emulsions, oil-water separation, adsorption, magnetic nanoparticles

Procedia PDF Downloads 107

Authors: Tuaumelsan Shumye Gebre, Shimelis Admassu Emire, Simon Okomo Aloo, Ramachandran Chelliah, Deog-Hwan Oh

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This study investigates the fermentation of cereal substrates, based on the Ethiopian traditional beverage borde, using probiotic strains of Pediococcus acidilactici WS07 and Saccharomyces cerevisiae AM18 used singly and in co-culture. The pH and titratable acidity, microbial growth dynamics, fermentable sugars profile, volatile organic compounds, total flavonoid content, total phenolic content, antioxidant activity, pancreatic lipase, and α-glucosidase inhibition were analyzed. The viability of every tested strain remained higher than 7 log CFU/mL, satisfying the requirements suggested for probiotic food items. The formation of organic acids is what caused the pH to decrease from roughly 6.6 to 3.8, yet this had no effect on the viability of the microorganisms. The fermentation process, involving P. acidilactici WS07 and S. cerevisiae AM18, led to the utilization of initial carbohydrates, production of organic acids, and generation of volatile compounds that enhance flavor and aroma. Ethanol and glycerol concentrations increased during fermentation, particularly in co-culture assays, contributing to the sensory qualities and stability of the beverages. The primary organic acids generated during fermentation were lactic and acetic acids. A total of 22 volatile substances, such as acids, alcohols, aldehydes, esters, ketones, and other substances, were found. Furthermore, the study demonstrates that fermentation of maize and sorghum with P. acidilactici WS07 and S. cerevisiae AM18 enhances the antioxidant activity and inhibition of pancreatic lipase and α-glucosidase, suggesting potential benefits in managing obesity and diabetes. Therefore, co-cultivating S. cerevisiae AM18 and P. acidilactici WS07 in cereal fermentation led to the successful production of probiotic drinks.

Keywords: probiotic beverage, Pediococcus acidilactici, Saccharomyces cerevisiae, volatile compounds

Procedia PDF Downloads 39

Authors: Khayra Zerrouki, Noureddine Djebli, Esra Eroglu, Afife Mat, Ozhan Gul

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Neurodegenerative diseases of the human brain comprise a variety of disorders that affect an increasing percentage of the population. Alzheimer’s disease (AD) is a complex, multifactorial, heterogeneous mental illness, which is characterized by an age-dependent loss of memory and an impairment of multiple cognitive functions, but this 10 last years it concerns the population most and most young. Hypericum perforatum has traditionally been used as an external anti-inflammatory and healing remedy for the treatment of swellings, wounds and burns, diseases of the alimentary tract and psychological disorders. It is currently of great interest due to new and important therapeutic applications. In this study, the chemical composition of methanolic extract of Hypericum perforatum (HPM) was analysed by using high performance liquid chromatography – diode array detector (HPLC-DAD). The in vitro antioxidant activity of HPM was evaluated by using several antioxidant tests. HSM exhibits inhibitory capacity against posphatidylcholine liposome peroxidation, induced with iron and ascorbic acid, scavenge DPPH and superoxide radicals and act as reductants. The cytotoxic activity of HSM was also determined by using MTT cell viability assay on HeLa and NRK-52E cell lines. The in vivo activity studies in Swiss mice were determined by using behavioral, memory tests and histological study. According to tests results HPM that may be relevant to the treatment of cognitive disorders. The results of chemical analysis showed a hight level of hyperforin and quercitin that had an important antioxidant activity proved in vitro with the DPPH, anti LPO and SOD; this antioxidant activity was confirmed in vivo after the non-toxic results by means of improvement in behavioral and memory than the reducing shrunken in pyramidal cells of mice brains.

Keywords: AlCl3, alzheimer, mice, neuroprotective, neurotoxicity, phytotherapy

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Authors: Staniek Halina

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The inappropriate trace elements supply such as iron(III) and chromium(III) may be risk factors of many metabolic disorders (e.g., anemia, diabetes, as well cause toxic effect). However, little is known about their mutual interactions and their impact on these disturbances. The effects of Cr(III) supplementation with a deficit or excess supply of Fe(III) in vivo conditions are not known yet. The objective of the study was to investigate the combined effect of different Fe(III) levels in the diet and simultaneous Cr(III) supplementation on the Ca distribution in organs in healthy rats. The assessment was based on a two-factor (2x3) experiment carried out on 54 female Wistar rats (Rattus norvegicus). The animals were randomly divided into 9 groups and for 6 weeks, they were fed semi-purified diets AIN-93 with three different Fe(III) levels in the diet as a factor A [control (C) 45 mg/kg (100% Recommended Daily Allowance for rodents), deficient (D) 5 mg/kg (10% RDA), and oversupply (H) 180 mg/kg (400% RDA)]. The second factor (B) was the simultaneous dietary supplementation with Cr(III) at doses of 1, 50 and 500 mg/kg of the diet. Iron(III) citrate was the source of Fe(III). The complex of Cr(III) with propionic acid, also called Cr₃ or chromium(III) propionate (CrProp), was used as a source of Cr(III) in the diet. The Ca content of analysed samples (liver, kidneys, spleen, heart, and femur) was determined with the Atomic Absorption Spectrometry (AAS) method. It was found that different dietary Fe(III) supply as well as Cr(III) supplementation independently and in combination influenced Ca metabolism in healthy rats. Regardless of the supplementation of Cr(III), the oversupply of Fe(III) (180 mg/kg) decreased the Ca content in the liver and kidneys, while it increased the Ca saturation of bone tissue. High Cr(III) doses lowered the hepatic Ca content. Moreover, it tended to decrease the Ca content in the kidneys and heart, but this effect was not statistically significant. The combined effect of the experimental factors on the Ca content in the liver and the femur was observed. With the increase in the Fe(III) content in the diet, there was a decrease in the Ca level in the liver and an increase in bone saturation, and the additional Cr(III) supplementation intensified those effects. The study proved that the different Fe(III) content in the diet, independently and in combination with Cr(III) supplementation, affected the Ca distribution in organisms of healthy rats.

Keywords: calcium, chromium(III), iron(III), rats, supplementation

Procedia PDF Downloads 199

Authors: Wasim Sajjad, Manzoor Ahmad, Sundas Qadir, Muhammad Rafiq, Fariha Hasan, Richard Tehan, Kerry L. McPhail, Aamer Ali Shah

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Aim: This study aims to investigate the possible radiation protective role of a compatible solute in the tolerance of radio-halophilic bacterium against stresses, like desiccation and exposure to ionizing radiation. Methods and Results: Nine different radio-resistant bacteria were isolated from desert soil, where strain WMA-LM19 was chosen for detailed studies on the basis of its high tolerance for ultraviolet radiation among all these isolates. 16S rRNA gene sequencing indicated that the bacterium was closely related to Stenotrophomonas sp. (KT008383). A bacterial milking strategy was applied for extraction of intracellular compatible solutes in 70% (v/v) ethanol, which were purified by high-performance liquid chromatography (HPLC). The compound was characterized as ectoine by 1H and 13C nuclear magnetic resonance (NMR), and mass spectrometry (MS). Ectoine demonstrated more efficient preventive activity (54.80%) to erythrocyte membranes and also inhibited oxidative damage to proteins and lipids in comparison to the standard ascorbic acid. Furthermore, a high level of ectoine-mediated protection of bovine serum albumin against ionizing radiation (1500-2000 Jm-2) was observed, as indicated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis. Conclusion: The results indicated that ectoine can be used as a potential mitigator and radio-protective agent to overcome radiation- and salinity-mediated oxidative damage in extreme environments. Significance and Impact of the Study: This study shows that ectoine from radio-halophiles can be used as a potential source in topical creams as sunscreen. The investigation of ectoine as UV protectant also changes the prospective that radiation resistance is specific only to molecular adaptation.

Keywords: ectoine, anti-oxidant, stenotrophomonas sp., ultraviolet radiation

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Authors: Rafik Balti, Mohamed Ben Mansour, Abdellah Arhaliass, Anthony Masse

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Unfortunately, shrimps are highly perishable and they start deteriorating immediately after death owing to their high water content and nutritional components. Currently, there has been an increasing interest in bioactive edible films and coatings to preserve the freshness and quality of foods. In this study, active edible coatings from microalgal exopolysaccharides (EPS) enriched with different concentrations of Red Seaweed Extract (RSE) (0.5, 1 and 1.5 % (w/v)) were developed and their effects on the quality changes of white shrimp during refrigerated storage (4 ± 1 °C) were examined over a period of 8 days. The control and the coated shrimp samples were analyzed periodically for microbiological (total viable bacteria, psychrotrophic bacteria, and enterobacteriaceae counts), chemical (pH, TVB-N, TMA-N, PV, TBARS), textural and sensory characteristics. The results indicated that the coating with a mixture of EPS and RSE could significantly decrease the total volatile basic nitrogen (TVB-N), trimethylamine (TMA) and thiobarbituric acid reactive substances (TBARS) (p < 0.05). With storage, EPS coatings containing RSE at both levels (1 and 1.5 %) were more effective in inhibiting the microbial species studied, specially psychrotrophic bacteria. Also, EPS + RSE coated samples had lower polyphenol oxidase (PPO) activity and lipid oxidation (p < 0.05) toward the end of storage. Textural and color properties of coated shrimp were generally more acceptable. Sensory scores indicated no significant changes in all samples during storage. The obtained results indicate that the edible EPS coating solutions enriched with RSE have noticeable effects on the quality and shelf life of shrimps when compared to control group. Finally, the present work demonstrates the effectiveness of EPS enriched coatings, offering a promising alternative to preserve more better the quality characteristics and to extend the shelf life of shrimp during the refrigerated storage

Keywords: active coating, exopolysaccharides, red seaweed, refrigerated storage, white shrimp

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Authors: Mohammad Faheem, M. Tabish Rehman, Mohd Danishuddin, Asad U. Khan

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The worldwide dissemination of CTX-M type β-lactamases is a threat to human health. Previously, we have reported the spread of blaCTX-M-15 gene in different clinical strains of Enterobacteriaceae from the hospital settings of Aligarh in north India. In view of the varying resistance pattern against cephalosporins and other β-lactam antibiotics, we intended to understand the correlation between MICs and catalytic activity of CTX-M-15. In this study, steady-state kinetic parameters and MICs were determined on E. coli DH5α transformed with blaCTX-M-15 gene that was cloned from Enterobacter cloacae (EC-15) strain of clinical background. The effect of conventional β-lactamase inhibitors (clavulanic acid, sulbactam and tazobactam) on CTX-M-15 was also studied. We have found that tazobactam is the best among these inhibitors against CTX-M-15. The inhibition characteristic of tazobactam is defined by its very low IC50 value (6 nM), high affinity (Ki = 0.017 µM) and better acylation efficiency (k+2/K9 = 0.44 µM-1s-1). It forms an acyl-enzyme covalent complex, which is quite stable (k+3 = 0.0057 s-1). Since increasing resistance has been reported against conventional b-lactam antibiotic-inhibitor combinations, we aspire to design a non-b-lactam core containing b-lactamase inhibitor. For this, we screened ZINC database and performed molecular docking to identify a potential non-β-lactam based inhibitor (ZINC03787097). The MICs of cephalosporin antibiotics in combination with this inhibitor gave promising results. Steady-state kinetics and molecular docking studies showed that ZINC03787097 is a reversible inhibitor which binds non-covalently to the active site of the enzyme through hydrogen bonds and hydrophobic interactions. Though, it’s IC50 (180 nM) is much higher than tazobactam, it has good affinity for CTX-M-15 (Ki = 0.388 µM). This study concludes that ZINC03787097 compound can be used as seed molecule to design more efficient non-b-lactam containing b-lactamase inhibitor that could evade pre-existing bacterial resistance mechanisms.

Keywords: ESBL, non-b-lactam-b-lactamase inhibitor, bioinformatics, biomedicine

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Authors: M. L. Mangena, N. Mokoena, K. Rashamuse, M. G. Tlou

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Tertiary alcohol ester (TAE) hydrolases are a group of esterases (EC 3.1.1.-) that catalyze the kinetic resolution of TAEs and as a result, they are sought-after for the production of optically pure tertiary alcohols (TAs) which are useful as building blocks for number biologically active compounds. What sets these enzymes apart is, the presence of a GGG(A)X-motif in the active site which appears to be the main reason behind their activity towards the sterically demanding TAEs. The genome of Pseudomonas syringae pv. maculicola (Psm) comprises a multitude of genes that encode esterases. We therefore, hypothesize that some of these genes encode TAE hydrolases. In this study, Psm was screened for TAE hydrolase activity using the linalyl acetate (LA) plate assay and a positive reaction was observed. As a result, the genome of Psm was screened for esterases with a GGG(A)X-motif using the motif search tool and two potential TAE hydrolase genes (PsmEST1 and 2, 1100 and 1000bp, respectively) were identified, PsmEST1 was amplified by PCR and the gene sequenced for confirmation. Analysis of the sequence data with the SingnalP 4.1 server revealed that the protein comprises a signal peptide (22 amino acid residues) on the N-terminus. Primers specific for the gene encoding the mature protein (without the signal peptide) were designed such that they contain NdeI and XhoI restriction sites for directional cloning of the PCR products into pET28a. The gene was expressed in E. coli JM109 (DE3) and the clones screened for TAE hydrolase activity using the LA plate assay. A positive clone was selected, overexpressed and the protein purified using nickel affinity chromatography. The activity of the esterase towards LA was confirmed using thin layer chromatography.

Keywords: hydrolases, tertiary alcohol esters, tertiary alcohols, screening, Pseudomonas syringae pv., maculicola genome, esterase activity, linalyl acetate

Procedia PDF Downloads 356