Search results for: protein surfactant interaction

Authors: Morgane Chatard, Clémentine Puech, Nathalie Perek, Frédéric Roche

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Several neurological disorders are linked to repeated hypoxia. The impact of such repeated hypoxic stress, on endothelial cells function of the blood-brain barrier (BBB) is little studied in the literature. Indeed, the study of hypoxic stress in cellular pathways is complex using hypoxia exposure because HIF 1α (factor induced by hypoxia) has a short half life. Our study presents an innovative induction method of repeated hypoxic stress, more reproducible, which allows us to study its impacts on an in vitro contact co-culture BBB model. Repeated hypoxic stress was induced by hydralazine (a mimetic agent of hypoxia pathway) during two hours and repeated during 24 hours. Then, BBB integrity was assessed by permeability measurements (transendothelial electrical resistance and membrane permeability), tight junction protein expressions (cell-ELISA and confocal microscopy) and by studying expression and activity of efflux transporters. First, this study showed that repeated hypoxic stress leads to a BBB’s dysfunction illustrated by a significant increase in permeability. This loss of membrane integrity was linked to a significant decrease of tight junctions’ protein expressions, facilitating a possible transfer of potential cytotoxic compounds in the brain. Secondly, we demonstrated that brain microvascular endothelial cells had set-up defence mechanism. These endothelial cells significantly increased the activity of their efflux transporters which was associated with a significant increase in their expression. In conclusion, repeated hypoxic stress lead to a loss of BBB integrity with a decrease of tight junction proteins. In contrast, endothelial cells increased the expression of their efflux transporters to fight against cytotoxic compounds brain crossing. Unfortunately, enhanced efflux activity could also lead to reducing pharmacological drugs delivering to the brain in such hypoxic conditions.

Keywords: BBB model, efflux transporters, repeated hypoxic stress, tigh junction proteins

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Authors: Mark Bezner, Shani Deri, Tom Trigano, Kfir Ben-Harush

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Intermediate filament (IF) proteins-based fibers are among the toughest fibers in nature, as was shown by native hagfish slime threads and by synthetic fibers that are based on type V IF-proteins, the nuclear lamins. It is assumed that their mechanical performance stems from two major factors: (1) the transition from elastic -helices to stiff-sheets during tensile load; and (2) the specific organization of the coiled-coil proteins into a hierarchical network of nano-filaments. Here, we investigated the interrelationship between these two factors by using wet-spun fibers based on C. elegans (Ce) lamin. We found that Ce-lamin fibers, whether assembled in aqueous or alcoholic solutions, had the same nonlinear mechanical behavior, with the elastic region ending at ~5%. The pattern of the transition was, however, different: the ratio between -helices and -sheets/random coils was relatively constant until a 20% strain for fibers assembled in an aqueous solution, whereas for fibers assembled in 70% ethanol, the transition ended at a 6% strain. This structural phenomenon in alcoholic solution probably occurred through the transition between compacted and extended conformation of the random coil, and not between -helix and -sheets, as cycle analyses had suggested. The different transition pattern can also be explained by the different higher order organization of Ce-lamins in aqueous or alcoholic solutions, as demonstrated by introducing a point mutation in conserved residue in Ce-lamin gene that alter the structure of the Ce-lamins’ nano-fibrils. In addition, biomimicking the layered structure of silk and hair fibers by coating the Ce-lamin fiber with a hydrophobic layer enhanced fiber toughness and lead to a reversible transition between -helix and the extended conformation. This work suggests that different hierarchical structures, which are formed by specific assembly conditions, lead to diverse secondary structure transitions patterns, which in turn affect the fibers’ mechanical properties.

Keywords: protein-based fibers, intermediate filaments (IF) assembly, toughness, structure-property relationships

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Authors: Ufaith Qadri, M Marouf Wani

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In this paper, the performance and emission characteristics of a Single Cylinder Spark Ignition engine have been investigated. The research is based on micro emulsion application as fuel in a gasoline engine. We have analyzed many micro emulsion compositions in various proportions, for predicting the performance of the Spark Ignition engine. This new technology of fuel modifications is emerging very rapidly as lot of research is going on in the field of micro emulsion fuels in Compression Ignition engines, but the micro emulsion fuel used in a Gasoline engine is very rare. The use of micro emulsion as fuel in a Spark Ignition engine is virtually unexplored. So, our main goal is to see the performance and emission characteristics of micro emulsions as fuel, in Spark Ignition engines, and finding which composition is more efficient. In this research, we have used various micro emulsion fuels whose composition varies for all the three blends, and their performance and emission characteristic were predicted in AVL Boost software. Conventional Gasoline fuel 90%, 80% and 85% were blended with co-surfactant Ethanol in different compositions, and water was used as an additive for making it crystal clear transparent micro emulsion fuel, which is thermodynamically stable. By comparing the performances of engines, the power has shown similarity for micro emulsion fuel and conventional Gasoline fuel. On the other hand, Torque and BMEP shows increase for all the micro emulsion fuels. Micro emulsion fuel shows higher thermal efficiency and lower Specific Fuel Consumption for all the compositions as compared to the Gasoline fuel. Carbon monoxide and Hydro carbon emissions were also measured. The result shows that emissions decrease for all the composition of micro emulsion fuels, and proved to be the most efficient fuel both in terms of performance and emission characteristics.

Keywords: AVL Boost, emissions, microemulsions, performance, Spark Ignition (SI) engine

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Authors: O. Adjroud

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The present study was conducted to investigate the interaction between selenium (Se) and chloride nickel (NiCl2) on energy profiles in Wistar albino preimplanted rats. NiCl2 was given on day 3 of pregnancy either in distilled drinking water at a dose of 20 mg/L/day for 16 consecutive days or as a single subcutaneous (s.c.) dose of 25, 50, or 100 mg/kg. Se was given as a s.c. injection (0.3 mg/kg) together with the higher dose (100 mg/kg) of NiCl2. Changes in energy profiles were evaluated in treated and control groups on days 5 and 20 of gestation. NiCl2 s.c. induced a significant increase in plasma glucose on day 20 of pregnancy. NiCl2 s.c. induced on day 5 and 20 of gestation a significant decrease in plasma triglycerides, with the higher dose. This decrease was maintained at day 20 of gestation with doses of 50 mg /kg. In addition, NiCl2 s.c. caused on day 5 of gestation a significant decrease in plasma total cholesterol with the low and medium doses. The pretreatment with Se reversed the effects of NiCl2 on plasma glucose, total cholesterol and triglycerides levels. NiCl2 administered in the drinking water augmented significantly the plasma triglycerides and total cholesterol levels and slighty the plasma glucose on day 20 of gestation, while on day 5 of gestation NiCl2 s.c. Induced a significant decrease in cholesterol. Three doses of NiCl2 (sc) induced severe alterations in liver and architecture which are markedly improved by Selenium. These results suggested that selenium has protective effects on energy profiles against the toxicity induced by NiCl2 administered subcutaneously in preimplanted rats.

Keywords: hepatotoxicity, nickel chloride, preimplanted rat, biochemical parameters

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Authors: Syeda Hira, Muhammad Gulfraz

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Background: Liver diseases cause serious health issues. Plants contain active compounds that significantly help in the treatment of various diseases. Ficus carica is traditionally used for the treatment of liver diseases. The purpose of the present study was the isolation and identification of active components from F.carica leaves which are responsible for hepatoprotective activity. Methods: The study was designed to identify the most active hepatoprotective sub-fraction from ethyl acetate fraction of Ficus carica by in vitro study and evaluation of its in vivo hepatoprotective effect in animal models. Ethyl acetate fraction was subjected to column, and a total of eight sub-fractions were obtained. In vitro, the hepatoprotective effect of all sub-fractions was determined on HepG2 cell lines. Toxicity was induced by CCl₄ (Carbon tetrachloride), and silymarin was used as a positive control. On the basis of the results, the most active sub-fraction was subjected to LC-MS and FT-IR analysis for the identification of bioactive compounds. In vivo, the hepatoprotective effect was determined in mice. Toxicity was induced by CCl₄; at the end of the experiment, biochemical parameters such as ALT, AST, ALP, bilirubin, and total protein were estimated in serum. Histopathology of liver tissues was also done. Results: Sub-fraction FVI exhibited significant (P<0.05) hepatoprotective activity as compared to other sub-fractions, which was almost similar to the standard drug silymarin. Six known bioactive compounds were identified from this sub-fraction after LC-MS analysis. In vivo, the hepatoprotective activity of sub-fraction FVI was evaluated in CCl₄-induced toxicated mice. Administration of CCl₄ significantly increased level of ALT (Alanine transaminase), AST (Aspartate aminotransferase), ALP (Alkaline phosphatase), and bilirubin and decreased the total protein. Treatment with sub-fraction FVI significantly (p<0.05) reversed the level of these biomarkers toward normal at both doses of 25 mg/kg and 50 mg/kg. Conclusion: Our findings confirmed the hepatoprotective effect of ethyl acetate fraction of F.carica. It could be a good candidate for the development of a natural hepatoprotective drug; pre-clinical investigation on ethyl acetate fraction is recommended.

Keywords: Ficus carica, hepatoprotective, CCl₄, bioactive compounds, liver markers

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Authors: Zakia Belhadj, Bing He, Hua Zhang, Xueqing Wang, Wenbing Dai, Qiang Zhang

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Mononuclear phagocyte system (MPS) forms an abominable obstacle hampering the tumor delivery efficiency of nanoparticles. Passively targeted nanocarriers have received clinical approval over the past 20 years. However, none of the actively targeted nanocarriers have entered clinical trials. Thus it is important to endue effective targeting ability to actively targeted approaches by overcoming biological barriers to nanoparticle drug delivery. Here, it presents that an Esomeprazole-based preconditioning strategy for regulating nanocarrier-MPS interaction to substantially prolong circulation time and enhance tumor targeting of nanoparticles. In vitro, the clinically approved proton pump inhibitor Esomeprazole “ESO” was demonstrated to reduce interactions between macrophages and subsequently injected targeted vesicles by interfering with their lysosomal trafficking. Of note, in vivo studies demonstrated that ESO pretreatment greatly decreased the liver and spleen uptake of c(RGDm7)-modified vesicles, highly enhanced their tumor accumulation, thereby provided superior therapeutic efficacy of c(RGDm7)-modified vesicles co-loaded with Doxorubicin (DOX) and Gefitinib (GE). This MPS-preconditioning strategy using ESO provides deeper insights into regulating nanoparticles interaction with the phagocytic system and enhancing their cancer cells' accessibility for anticancer therapy.

Keywords: esomeprazole (ESO), mononuclear phagocyte system (MPS), preconditioning strategy, targeted lipid vesicles

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Authors: Sameh A. S. Thabit Alariqi

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Our previous studies aimed to investigate the biodegradation of γ-sterilized polyolefins in composting and microbial culture environments at different doses and γ-dose rates. It was concluded from the previous studies that the pretreatment of γ-irradiation can accelerate the biodegradation of neat polymer matrix in biotic conditions significantly. A similar work was carried out to study the stabilization of γ-sterilized polyolefins using different mixtures of stabilizers which are approved for food-contact applications. Ethylene-propylene (EP) copolymer has been melt-mixed with hindered amine stabilizers (HAS), phenolic antioxidants and hydroperoxide decomposers. Results were discussed by comparing the stabilizing efficiency, combination and consumption of stabilizers and the synergistic and antagonistic effects was explained through the interaction between the stabilizers. In this attempt, we have aimed to study the influence of the synergistic and antagonistic mixtures of oligomeric stabilizers on the biodegradation of the γ-irradiated polyolefins in composting and microbial culture. Neat and stabilized films of EP copolymer irradiated under γ-radiation and incubated in compost and fungal culture environments. The changes in functional groups, surface morphology, mechanical properties and intrinsic viscosity in polymer chains were characterized by FT-IR spectroscopy, SEM, instron, and viscometric measurements respectively. Results were discussed by comparing the effect of different stabilizers, stabilizers mixtures on the biodegradation of the γ-irradiated polyolefins. It was found that the biodegradation significantly depends on the components of stabilization system, mobility, interaction, and consumption of stabilizers.

Keywords: biodegradation, γ-irradiation, polyolefins, stabilization

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

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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: Mohadese Hashemi, Elham Akhoundi Kharanaghi, Fatemeh Haghiralsadat, Mojgan Yazdani, Omid Javani, Mahboobe Sharafodini, Davood Rajabi

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Niosomes are non-ionic surfactant vesicles in aqueous media resulting in closed bilayer structures that can be used as carriers of hydrophilic and hydrophobic compounds. The use of niosomes for encapsulation of essential oils (EOs) is an attractive new approach to overcome their physicochemical stability concerns include sensibility to oxygen, light, temperature, and volatility, and their reduced bioavailability which is due to low solubility in water. EOs are unstable and fragile volatile compounds which have strong interest in pharmaceutical due to their medicinal properties such as antiviral, anti-inflammatory, antifungal, and antioxidant activities without side effects. Trachyspermum copticum (ajwain) is an annual aromatic plant with important medicinal properties that grows widely around Mediterranean region and south-west Asian countries. The major components of the ajwain oil were reported as thymol, γ-terpinene, p-cymene, and carvacrol which provide antimicrobial and antioxidant activity. The aim of this work was to formulate ajwain essential oil-loaded niosomes to improve water solubility of natural product and evaluate its physico-chemical features and stability. Ajwain oil was obtained through steam distillation using a clevenger-type apparatus and GC/MS was applied to identify the main components of the essential oil. Niosomes were prepared by using thin film hydration method and nanoparticles were characterized for particle size, dispersity index, zeta potential, encapsulation efficiency, in vitro release, and morphology.

Keywords: trachyspermum copticum, ajwain, niosome, essential oil, encapsulation

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Authors: Rita Greco, Giuseppe Carlo Marano

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Chloride induced reinforcement corrosion is widely accepted to be the most frequent mechanism causing premature degradation of reinforced concrete members, whose economic and social consequences are growing up continuously. Prevention of these phenomena has a great importance in structural design, and modern Codes and Standard impose prescriptions concerning design details and concrete mix proportion for structures exposed to different external aggressive conditions, grouped in environmental classes. This paper focuses on reinforced concrete columns load carrying capacity degradation over time due to chloride induced steel pitting corrosion. The structural element is considered to be exposed to marine environment and the effects of corrosion are described by the time degradation of the axial-bending interaction diagram. Because chlorides ingress and consequent pitting corrosion propagation are both time-dependent mechanisms, the study adopts a time-variant predictive approach to evaluate the residual strength of corroded reinforced concrete columns at different lifetimes. Corrosion initiation and propagation process is modelled by taking into account all the parameters, such as external environmental conditions, concrete mix proportion, concrete cover and so on, which influence the time evolution of the corrosion phenomenon and its effects on the residual strength of RC columns.

Keywords: pitting corrosion, strength deterioration, diffusion coefficient, surface chloride concentration, concrete structures, marine environment

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Authors: Radovan Čobanović, Milica Rankov Šicar

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New attitudes about nutrition impose new styles, and therefore a neNew attitudes about nutrition impose new styles, and therefore a new kind of food. The goal of our work was to define the shelf life of new extruded soya product with minimum 65% of protein based on the analyses. According to the plan it was defined that a certain quantity of the same batch of new product (soybean flakes) which had predicted shelf life of 2 years had to be stored for 24 months in storage and analyzed at the beginning and end of sustainability plan on instrumental analyses (heavy metals, pesticides and mycotoxins) and every month on sensory analyses (odor, taste, color, consistency), microbiological analyses (Salmonella spp., Escherichia coli, Enterobacteriaceae, sulfite-reducing clostridia, Listeria monocytogenes), chemical analyses (protein, ash, fat, crude cellulose, granulation) and at the beginning on GMO analyses. All analyses were tested according to: sensory analyses ISO 6658, Salmonella spp ISO 6579, Escherichia coli ISO 16649-2, Enterobacteriaceae ISO 21528-2, sulfite-reducing clostridia ISO 15213 and Listeria monocytogenes ISO 11290-2, chemical and instrumental analyses Serbian ordinance on the methods of physico-chemical analyses and GMO analyses JRC Compendium. The results obtained after the analyses which were done according to the plan during the 24 months indicate that are no changes of products concerning both sensory and chemical analyses. As far as microbiological results are concerned Salmonella spp was not detected and all other quantitative analyses showed values <10 cfu/g. The other parameters for food safety (heavy metals, pesticides and mycotoxins) were not present in analyzed samples and also all analyzed samples were negative concerning genetic testing. On the basis of monitoring the sample under defined storage conditions and analyses of quality control, GMO analyses and food safety of the sample during the shelf within two years, the results showed that all the parameters of the sample during defined period is in accordance with Serbian regulative so that indicate that predicted shelf life can be adopted.w kind of food. The goal of our work was to define the shelf life of new extruded soya product with minimum 65% of protein based on the analyses. According to the plan it was defined that a certain quantity of the same batch of new product (soybean flakes) which had predicted shelf life of 2 years had to be stored for 24 months in storage and analyzed at the beginning and end of sustainability plan on instrumental analyses (heavy metals, pesticides and mycotoxins) and every month on sensory analyses (odor, taste, color, consistency), microbiological analyses (Salmonella spp., Escherichia coli, Enterobacteriaceae, sulfite-reducin clostridia, Listeria monocytogenes), chemical analyses (protein, ash, fat, crude cellulose, granulation) and at the beginning on GMO analyses. All analyses were tested according: sensory analyses ISO 6658, Salmonella spp ISO 6579, Escherichia coli ISO 16649-2, Enterobacteriaceae ISO 21528-2, sulfite-reducing clostridia ISO 15213 and Listeria monocytogenes ISO 11290-2, chemical and instrumental analyses Serbian ordinance on the methods of physico-chemical analyses and GMO analyses JRC Compendium. The results obtained after the analyses which were done according to the plan during the 24 months indicate that are no changes of products concerning both sensory and chemical analyses. As far as microbiological results are concerned Salmonella spp was not detected and all other quantitative analyses showed values <10 cfu/g. The other parameters for food safety (heavy metals, pesticides and mycotoxins) were not present in analyzed samples and also all analyzed samples were negative concerning genetic testing. On the basis of monitoring the sample under defined storage conditions and analyses of quality control, GMO analyses and food safety of the sample during the shelf within two years, the results showed that all the parameters of the sample during defined period is in accordance with Serbian regulative so that indicate that predicted shelf life can be adopted.

Keywords: extruded soya product, food safety analyses, GMO analyses, shelf life

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Authors: Alireza Gholipour, Mergen H. Ghayesh, Anthony Zander, Stephen J. Nicholls, Peter J. Psaltis

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A three-dimensional numerical model of an atherosclerotic coronary ‎artery is developed for the determination of high-risk situation and ‎hence heart attack prediction. Employing the finite element method ‎‎(FEM) using ANSYS, fluid-structure interaction (FSI) model of the ‎artery is constructed to determine the shear stress distribution as well ‎as the von Mises stress field. A flexible model for an atherosclerotic ‎coronary artery conveying pulsatile blood is developed incorporating ‎three-dimensionality, artery’s tapered shape via a linear function for ‎artery wall distribution, motion of the artery, blood viscosity via the ‎non-Newtonian flow theory, blood pulsation via use of one-period ‎heartbeat, hyperelasticity via the Mooney-Rivlin model, viscoelasticity ‎via the Prony series shear relaxation scheme, and micro-calcification ‎inside the plaque. The material properties used to relate the stress field ‎to the strain field have been extracted from clinical data from previous ‎in-vitro studies. The determined stress fields has potential to be used as ‎a predictive tool for plaque rupture and dissection.‎ The results show that stress concentration due to micro-calcification ‎increases the von Mises stress significantly; chance of developing a ‎crack inside the plaque increases. Moreover, the blood pulsation varies ‎the stress distribution substantially for some cases.‎

Keywords: atherosclerosis, fluid-structure interaction‎, coronary arteries‎, pulsatile flow

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Authors: Franziska Köhler-Dauner, Inka Mayer, Lara Hart, Ute Ziegenhain, Jörg M. Fegert

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Preventive isolation and social distancing strategies during the SARS-CoV-2 pandemic have confronted families with a variety of different restrictions and stresses. Especially during this stressful time, children need a stable parental home to avoid developmental disorders. Additional risk factors such as maternal childhood abuse and neglect (CM) experiences may influence mothers' psychosomatic health (pG) and children's physical well-being (kW) during times of increased stress. Our aim was to analyse the interaction between maternal CM, maternal pG and children's kW during the pandemic. Mothers from a well-documented birth cohort to study transgenerational transmission of CM, were included in an online 'pandemic' survey assessing maternal pG and children's physical health during the pandemic. Our mediation analysis showed a significant positive association between the extent of maternal CM experiences, mothers' psychosomatic symptoms and their children's kW. Maternal psychosomatic symptoms significantly mediate the interaction between CM and children's kW; the direct effect remains non-significant when maternal psychosomatic symptoms are included as mediators. Maternal CM appears to be a relevant risk factor for maternal pG and children's kW during the pandemic. Maternal CM experiences seem to influence the way parents cope with stressful situations and increase the risk of suffering from depressive symptoms. The latter also affect their children's kW. Our findings underline the importance of carefully assessing the specific situation of families with children and offering individually adapted help to help families survive the pandemic.

Keywords: pandemic, maternal health, child health, abuse, neglect, maternal experiences

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Authors: Govindan Sudha, Mathumitha Subramaniam, Alamelu Govindasamy, Sasikala Gunasekaran

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The aim of this study was to investigate the protective effect of Hypsizygus ulmarius polysaccharides (HUP) on reducing oxidative stress, cognitive impairment and neurotoxicity in D-galactose induced aging mice. Mice were subcutaneously injected with D-galactose (150 mg/kg per day) for 6 weeks and were administered HUP simultaneously. Aged mice receiving vitamin E (100 mg/kg) served as positive control. Chronic administration of D-galactose significantly impaired cognitive performance oxidative defence and mitochondrial enzymes activities as compared to control group. The results showed that HUP (200 and 400 mg/kg) treatment significantly improved the learning and memory ability in Morris water maze test. Biochemical examination revealed that HUP significantly increased the decreased activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (GR), glutathione-S-transferase (GST), mitochondrial enzymes-NADH dehydrogenase, malate dehydrogenase (MDH), isocitrate dehydrogenase (ICDH), Na+K+, Ca2+, Mg2+ATPase activities, elevated the lowered total anti-oxidation capability (TAOC), glutathione (GSH), vitamin C and decreased the raised acetylcholinesterase (AChE) activities, malondialdehyde (MDA), hydroperoxide (HPO), protein carbonyls (PCO), advanced oxidation protein products (AOPP) levels in brain of aging mice induced by D-gal in a dose-dependent manner. In conclusion, present study highlights the potential role of HUP against D-galactose induced cognitive impairment, biochemical and mitochondrial dysfunction in mice. In vitro studies on the effect of HUP on scavenging DPPH, ABTS, DMPD, OH radicals, reducing power, B-carotene bleaching and lipid peroxidation inhibition confirmed the free radical scavenging and antioxidant activity of HUP. The results suggest that HUP possesses anti-aging efficacy and may have potential in treatment of neurodegenerative diseases.

Keywords: aging, antioxidants, mushroom, neurotoxicity

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Authors: Manjusha Choudhary

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Objective: Cassia occidentalis Linn. belongs to Family Caesalpiniaceae is a common weed scattered from the foothills of Himalayas to West Bengal, South India, Burma, and Sri Lanka. It is used widely in folklore medicine in India as laxative, expectorant, analgesic, anti-malarial, hepatoprotective, relaxant, anti-inflammatory and antidiabetic. The present study was carried out to investigate the hypoglycaemic and hypolipidemic activities of ethanolic extract of Cassia occidentalis stem bark. Methods: Stem bark extract of Cassia occidentalis (SBCO) was administered orally at 250 and 500 mg/kg doses to normal and streptozotocin (STZ) induced type-2 diabetic mice. Various parameters like fasting blood glucose (FBG) level, serum cholesterol, high density lipoprotein (HDL) cholesterol, triglycerides (TG), total protein, urea, creatinine, serum glutamate oxaloacetate transaminase (SGOT), serum glutamate pyruvate transaminase (SGPT) levels and physical parameters like change in body weight, food intake, water intake were performed for the evaluation of antidiabetic effects. Results: Both the doses of extract caused a marked decrease in FBG levels in STZ induced type 2 diabetic mice. Administration of SBCO led to the decrease in the blood glucose, food intake, water intake, organ weight, SGOT, SGPT levels with significant value and increased the levels of TG, HDL cholesterol, creatinine, cholesterol, total protein with a significant value (p < 0.05-0.01). The decrease in body weight induced by STZ was restored to normal with a significant value (p < 0.01) at both doses. Conclusion: Present study reveals that SBCO possess potent hypoglycaemic and hypolipidemic activities and supports the folklore use of the stem bark of plant as antidiabetic agent.

Keywords: Cassia occidentalis, diabetes, folklore, herbs, hypoglycemia, streptozotocin

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Authors: Frida Carrillo Morales, Maria Maricela Carrasco Yépez, Saúl Rojas Hernández

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Naegleria fowleri is the causative agent of primary amebic meningoencephalitis, this disease is acute and fulminant that affects humans. It has been reported that despite the existence of therapeutic options against this disease, its mortality rate is 97%. Therefore, the need arises to have vaccines that confer protection against this disease and, in addition to developing adjuvants to enhance the immune response. In this regard, in our work group, we obtained a peptide designed from the membrane protein MP2CL5 of Naegleria fowleri called Smp145 that was shown to be immunogenic; however, it would be of great importance to enhance its immunological response, being able to co-administer it with a non-toxic adjuvant. Therefore, the objective of this work was to carry out the bioinformatic design of a peptide of the Naegleria fowleri membrane protein MP2CL5 conjugated with a non-toxic modified adjuvant from the native A1 structure of Cholera Toxin. For which different bioinformatics tools were used to obtain a model with a modification in amino acid 61 of the A1 subunit of the CT (CTA1), to which the Smp145 peptide was added and both molecules were joined with a 13-glycine linker. As for the results obtained, the modification in CTA1 bound to the peptide produces a reduction in the toxicity of the molecule in in silico experiments, likewise, the prediction in the binding of Smp145 to the receptor of B cells suggests that the molecule is directed in specifically to the BCR receptor, decreasing its native enzymatic activity. The stereochemical evaluation showed that the generated model has a high number of adequately predicted residues. In the ERRAT test, the confidence with which it is possible to reject regions that exceed the error values was evaluated, in the generated model, a high score was obtained, which determines that the model has a good structural resolution. Therefore, the design of the conjugated peptide in this work will allow us to proceed with its chemical synthesis and subsequently be able to use it in the mouse meningitis protection model caused by N. fowleri.

Keywords: immunology, vaccines, pathogens, infectious disease

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Authors: Maliheh Raji, Hossein Abolghasemi, Jaber Safdari, Ali Kargari

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Dysprosium is a rare earth element which is essential for many growing high-technology applications. Dysprosium along with neodymium plays a significant role in different applications such as metal halide lamps, permanent magnets, and nuclear reactor control rods preparation. The purification and separation of rare earth elements are challenging because of their similar chemical and physical properties. Among the various methods, membrane processes provide many advantages over the conventional separation processes such as ion exchange and solvent extraction. In this work, selective extraction and separation of dysprosium from aqueous solutions containing an equimolar mixture of dysprosium and neodymium by emulsion liquid membrane (ELM) was investigated. The organic membrane phase of the ELM was a nanofluid consisting of multiwalled carbon nanotubes (MWCNT), Span80 as surfactant, Cyanex 272 as carrier, kerosene as base fluid, and nitric acid solution as internal aqueous phase. Factors affecting separation of dysprosium such as carrier concentration, MWCNT concentration, feed phase pH and stripping phase concentration were analyzed using Taguchi method. Optimal experimental condition was obtained using analysis of variance (ANOVA) after 10 min extraction. Based on the results, using MWCNT nanofluid in ELM process leads to increase the extraction due to higher stability of membrane and mass transfer enhancement and separation factor of 6 for dysprosium over neodymium can be achieved under the optimum conditions. Additionally, demulsification process was successfully performed and the membrane phase reused effectively in the optimum condition.

Keywords: emulsion liquid membrane, MWCNT nanofluid, separation, Taguchi method

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Authors: Jinhee Kang, Kwangman Park, Ruhee Song, Suhee Kim, Do-Hyeon Yu, Kyoungseong Choi, Jinho Park

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Statement of the Problem: Animal blood components maintain homeostasis when animals are healthy, and changes in chemical composition of the blood and body fluids can be observed if animals have a disease. In particular, newborn calves are susceptible to disease and therefore hematologic tests and serum chemistry tests could become an important guideline to the diagnosis and the treatment of diseases. Diarrhea in newborn calves is the most damaging to cattle ranch, whether dairy or cattle fattening, and is a large part of calf atrophy and death. However, since the study on calf electrophoresis was not carried out, a survey analysis was conducted on it. Methodology and Theoretical Orientation: The calves were divided into healthy calves and disease (diarrhea) calves, and calves were classified by 1-14d, 15-28d, and more than 28d, respectively. The fecal state was classified by solid (0-value), semi-solid (1-value), loose (2-value) and watery (3-value). In the solid (0-value) and semi-solid (1-value) feces valuable pathogen was not detected, but loose (2-value) and watery (3-value) feces were detected. Findings: ALB, α-1, α-2, α-SUM, β and γ (Gamma) were examined by electrophoresis analysis of healthy calves and diarrhea calves. Test results showed that there were age differences between healthy calves and diarrheic calves. When we look at the γ-globulin at 1-14 days of age, we can see that the average calf of healthy calves is 16.8% and the average of diarrheal calves is 7.7%, when we look at the figures for the α-2 at 1-14 days, we found that healthy calves average 5.2% and diarrheal calves 8.7% higher than healthy cows. On α-1, 15-28 days, and after 28 days, healthy calves average 10.4% and diarrheal calves average 7.5% diarrhea calves were 12.6% and 12.4% higher than healthy calves. In the α-SUM, the healthy calves were 21.6%, 16.8%, and 14.5%, respectively, after 1-14 days, 15-28 days and 28 days. diarrheal calves were 23.1%, 19.5%, and 19.8%. Conclusion and Significance: In this study, we examined the electrophoresis results of healthy calves and diseased (diarrhea) calves, gamma globulin at 1-14 days of age were lower than those of healthy calves (diarrhea), indicating that the calf was unable to consume colostrum from the mother when it was a new calf. α-1, α-2, α-SUM may be associated with an acute inflammatory response as a result of increased levels of calves with diarrhea (diarrhea). Further research is needed to investigate the effects of acute inflammatory responses on additional calf-forming proteins. Information on the results of the electrophoresis test will be provided where necessary according to the item.

Keywords: alpha, electrophoretogram, serum protein, γ, gamma

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Authors: Asim Abbasi, Muhammad Sufyan

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The present experiment was carried out to examine the effects of silicon dioxide on tritrophic interaction of cotton, whitefly, and the predator Chrysoperla carnea. Population of whitefly was maintained on silicon treated and non-treated cotton for two generations in greenhouse net cages exposed to outside temperature and luminosity. The cotton was treated with silicon dioxide twice after 15 days intervals with 200 ppm concentration. A stock rearing of the natural predator was developed in the laboratory conditions. In the bioassay eggs of the predator all at the same age were individualized in glass petri plates that will be pierced with a pin to allow aeration and maintained in an incubator at 28 ± 2°C, 70 ± 10% relative humidity and 12h photo phase. Population of whitefly stayed on silicon treated, and non-treated cotton were offered to newly hatched chrysopid larvae until the end of the larval stage, assuring a permanent supply. Feeding preference of C. carnea along with longevity, survival of each instar larvae, pupation, adult emergence, and fecundity was checked. The results revealed that there was no significant difference in the feeding preference of C. carnea among both treatments. Durations of 1st and 2nd larval instar were also at par in both treatments. However overall longevity and adult emergence were a bit lower in silicon treated whitefly treatment. This may be due to the fact that silicon reduces the nutritional quality of host because of reduced whitefly feeding on silicon treated cotton. No significant difference in 1st and 2nd larval instars and then increased larval duration in later instars suggested that the effect of silicon treated host should be checked on more than 1 generation of C. carnea to get better findings.

Keywords: Chrysoperla carnea, silicon, tritrophic, whitefly

Procedia PDF Downloads 180

Authors: P. Annapurna, Cecil Ross, Sudhir Krishna, Sweta Srivastava

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Irradiation plays a pivotal role in cervical cancer treatment, however some tumors exhibit resistance to therapy while some exhibit relapse, due to better repair and enhanced resistance mechanisms operational in their cells. The present study aims to understand the signaling mechanism operational in resistance phenotype and in the present study we report the role of Rho GTPase associated protein kinase (ROCK) signaling in cervical carcinoma radio-resistance. ROCK signaling has been implicated in several tumor progressions and is important for DNA repair. Irradiation of spheroid cultures of SiHa cervical carcinoma derived cell line at 6Gy resulted in generation of resistant cells in vitro which had better clonogenic abilities and formed larger and more colonies, in soft agar colony formation assay, as compared to the non-irradiated cells. These cells also exhibited an enhanced motility phenotype. Cell cycle profiling showed the cells to be blocked in G2M phase with enhanced pCDC2 levels indicating onset of possible DNA repair mechanism. Notably, 3 days post-irradiation, irradiated cells showed increased ROCK2 translocation to the nucleus with enhanced protein expression as compared to the non-irradiated cells. Radio-sensitization of the resistant cells was enhanced using Y27632, an inhibitor to ROCK signaling. The treatment of resistant cells with Y27632 resulted in increased cell death upon further irradiation. This observation has been confirmed using inhibitory antibodies to ROCK1/2. Result show that both ROCK1/2 have a functional contribution in radiation resistance of cervical cancer cells derived from cell lines. Interestingly enrichment of stem like cells (Hoechst negative cells) was also observed upon irradiation and these cells were markedly sensitive to Y27632 treatment. Our results thus suggest the role of ROCK signaling in radio-resistance in cervical carcinoma. Further studies with human biopsies, mice models and mechanistic of ROCK signaling in the context of radio-resistance will clarify the role of this molecule further and allow for therapeutics development.

Keywords: cervical carcinoma, radio-resistance, ROCK signaling, cancer treatment

Procedia PDF Downloads 330

Authors: Ramon Omar Rosales-Espinoza

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Pile-to-pile mechanical connections are used if the depth of the soil layers with sufficient bearing strength exceeds the original (“leading”) pile length, with the additional pile segment being termed “extension” pile. Mechanical connectors permit a safe transmission of forces from leading to extension pile while meeting strength and serviceability requirements. Common types of connectors consist of an assembly of sleeve-type external couplers, bolts, pins, and other mechanical interlock devices that ensure the transmission of compressive, tensile, torsional and bending stresses between leading and extension pile segments. While welded connections allow for a relatively simple structural design, mechanical connections are advantageous over welded connections because they lead to shorter installation times and significant cost reductions since specialized workmanship and inspection activities are not required. However, common practices followed to design mechanical connectors neglect important aspects of the assembly response, such as stress concentration around pin/bolt holes, torsional stresses from the installation process, and interaction between the forces at the installation (torsion), service (compression/tension-bending), and removal stages (torsion). This translates into potentially unsatisfactory designs in terms of the ultimate and service limit states, exhibiting either reduced strength or excessive deformations. In this study, the experimental response under compressive forces of a type of mechanical connector is presented, in terms of strength, deformation and failure modes. The tests revealed that the type of connector used can safely transmit forces from pile to pile. Using the results from the compressive tests, an analysis model was developed using the finite element (FE) method to study the interaction of forces under installation and service stages of a typical mechanical connector. The response of the analysis model is used to identify potential areas for design optimization, including size, gap between leading and extension piles, number of pin/bolts, hole sizes, and material properties. The results show the design of mechanical connectors should take into account the interaction of forces present at every stage of their life cycle, and that the torsional stresses occurring during installation are critical for the safety of the assembly.

Keywords: piles, FEA, steel, mechanical connector

Procedia PDF Downloads 264

Authors: German Hernandez-Alonso, Antonio Lazcano, Arturo Becerra

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Until today, viruses remain controversial and poorly understood; about their origin, this problem represents an enigma and one of the great challenges for the contemporary biology. Three main theories have tried to explain the origin of viruses: regressive evolution, escaped host gene, and pre-cellular origin. Under the perspective of the escaped host gene theory, it can be assumed a cellular origin of viral components, like protein RNA-binding domains. These universal distributed RNA-binding domains are related to the RNA metabolism processes, including transcription, processing, and modification of transcripts, translation, RNA degradation and its regulation. In the case of viruses, these domains are present in important viral proteins like helicases, nucleases, polymerases, capsid proteins or regulation factors. Therefore, they are implicated in the replicative cycle and parasitic processes of viruses. That is why it is possible to think that those domains present low levels of divergence due to selective pressures. For these reasons, the main goal for this project is to create a catalogue of the RNA-binding domains found in all the available viral proteomes, using bioinformatics tools in order to analyze its evolutionary process, and thus shed light on the general virus evolution. ProDom database was used to obtain larger than six thousand RNA-binding domain families that belong to the three cellular domains of life and some viral groups. From the sequences of these families, protein profiles were created using HMMER 3.1 tools in order to find distant homologous within greater than four thousand viral proteomes available in GenBank. Once accomplished the analysis, almost three thousand hits were obtained in the viral proteomes. The homologous sequences were found in proteomes of the principal Baltimore viral groups, showing interesting distribution patterns that can contribute to understand the evolution of viruses and their host-virus interactions. Presence of cellular RNA-binding domains within virus proteomes seem to be explained by closed interactions between viruses and their hosts. Recruitment of these domains is advantageous for the viral fitness, allowing viruses to be adapted to the host cellular environment.

Keywords: bioinformatics tools, distant homology, RNA-binding domains, viral evolution

Procedia PDF Downloads 387

Authors: Nusrat Praween, Krishna Thej Pammi Guru, Palash Kumar Basu

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Designing non-invasive biosensors for cancer diagnosis is essential for developing an affordable and specific tool to measure cancer-related exosome biomarkers. Exosomes, released by healthy as well as cancer cells, contain valuable information about the biomarkers of various diseases, including cancer. Despite the availability of various isolation techniques, ultracentrifugation is the standard technique that is being employed. Post isolation, exosomes are traditionally exposed to detergents for extracting their proteins, which can often lead to protein degradation. Further to this, it is very essential to develop a sensing platform for the quantification of clinically relevant proteins in a wider range to ensure practicality. In this study, exosomes were immobilized on the Au Screen Printed Electrode (SPE) using EDC/NHS chemistry to facilitate binding. After immobilizing the exosomes on the screen-printed electrode (SPE), we investigated the impact of the electric field by applying various voltages to induce exosome lysis and release their contents. The lysed solution was used for sensing TSG101, a crucial biomarker associated with various cancers, using both faradaic and non-faradaic electrochemical impedance spectroscopy (EIS) methods. The results of non-faradaic and faradaic EIS were comparable and showed good consistency, indicating that non-faradaic sensing can be a reliable alternative. Hence, the non-faradaic sensing technique was used for label-free quantification of the TSG101 biomarker. The results were validated using ELISA. Our electrochemical immunosensor demonstrated a consistent response of TSG101 from 125 pg/mL to 8000 pg/mL, with a detection limit of 0.125 pg/mL at room temperature. Additionally, since non-faradic sensing is label-free, the ease of usage and cost of the final sensor developed can be reduced. The proposed immunosensor is capable of detecting the TSG101 protein at low levels in healthy serum with good sensitivity and specificity, making it a promising platform for biomarker detection.

Keywords: biosensor, exosomes isolation on SPE, electric field lysis of exosome, EIS sensing of TSG101

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Authors: Amany Ragab, Nashwa Khairat Abousamra, Omayma Saleh, Asmaa Higazy

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Insulin resistance syndrome has been shown to be associated with many coagulation and fibrinolytic proteins and these associations suggest that some coagulation and fibrinolytic proteins have a role in atherothrombotic disorders. This study was conducted to determine the levels of some of the haemostatic parameters in subjects having metabolic syndrome and to correlate these values with the anthropometric and metabolic variables associated with this syndrome. The study included 46 obese non diabetic subjects of whom 28 subjects(group1) fulfilled the ATP III criteria of the metabolic syndrome and 18 subjects (group2) did not have metabolic syndrome as well as 14 lean subjects (group 3) of matched age and sex as a control group. Clinical and laboratory evaluation of the study groups stressed on anthropometric measurements (weight, height, body mass index, waist circumference, and sagittal abdominal diameter), blood pressure, and laboratory measurements of fasting plasma glucose, fasting insulin, serum lipids, tissue plasminogen activator (t-PA), antithrombin III activity (ATIII), protein C and von Willebrand factor (vWf) antigen. There was significant increase in the concentrations of t-PA and vWf antigens in subjects having metabolic syndrome (group 1) in comparison to the other groups while there were non-significant changes in the levels of protein C antigen and AT III activity. Both t-PA and vWf showed significant correlation with HOMA-IR as a measure of insulin sensitivity. The t-PA showed also significant correlation with most of the variables of metabolic syndrome including waist circumference, BMI, systolic blood pressure, fasting plasma glucose, fasting insulin, and HDL cholesterol. On the other hand, vWf showed significant correlations with fasting plasma glucose, fasting insulin and sagital abdominal diameter, with non-significant correlations with the other variables. Haemostatic and fibrinolytic parameters should be included in the features and characterization of the insulin resistance syndrome. t-PA and vWf antigens concentrations were increased in subjects with metabolic syndrome and correlated with the HOMA-IR measure of insulin sensitivity. Taking into consideration that both t-PA and vWf are mainly released from vascular endothelium, these findings could be an indicator of endothelial dysfunction in that group of subjects.

Keywords: insulin resistance, obesity, metabolic syndrome, coagulation

Procedia PDF Downloads 137

Authors: Hassan Sadeghi Naeini, Mozhgan Sabzehparvar, Mahdiye Jafarnezhad, Neda Brumandi, Mohammad Parsa Sabzehparvar

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This study aims to discover the relationship between the factors of aesthetics, user experience, and social sustainability concerning the design of street furniture and the impact of these factors on the emotional arousal of citizens to encourage and make them prefer to use street furniture. The method used in this research included extracting indicators related to each of the factors of aesthetics, user experience, and social sustainability from the articles and then selecting indicators related to the purpose of the research in consultation with industrial design experts and architects. Finally, 9 variables for aesthetics, 7 variables for user experience, and 5 variables for evaluating social sustainability were selected. To identify the effect of each of these factors on street furniture and to recognize their relationship with each other. A 10-scale prioritization questionnaire, from 1, the least amount of importance, to 10, the most amount of importance, was answered by architects and industrial designers on the “Pors Line” online platform for three consecutive weeks, and a total of 82 people answered the questionnaire. The results showed that by using aesthetic factors in the design of street furniture and having a positive impact on users’ experience of using the product, we could expect the occurrence of behavioral factors, such as creating constructive interaction and product acceptance so that the satisfaction of the user in the use of street furniture and optimal interaction in the urban environment is formed, followed by that, the requirements of social sustainability will be met.

Keywords: visual aesthetic, user experience, social sustainability, street furniture

Procedia PDF Downloads 95

Authors: Indu Chauhan, Bhupendra S. Butola, Paritosh Mohanty

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It is very well known that properties of material changes as their size approach to nanoscale level due to the high surface area to volume ratio. However, in last few decades, a tenet ‘structure dictates function’ is quickly being adopted by researchers working with nanomaterials. The design and exploitation of nanoparticles with tailored shape and size has become one of the primary goals of materials science researchers to expose the properties of nanostructures. To date, various methods, including soft/hard template/surfactant assisted route hydrothermal reaction, seed mediated growth method, capping molecule-assisted synthesis, polyol process, etc. have been adopted to synthesize the nanostructures with controlled size and shape and monodispersity. However controlling the shape and size of nanoparticles is an ultimate challenge of modern material research. In particular, many efforts have been devoted to rational and skillful control of hierarchical and complex nanostructures. Thus in our research work, role of cellulose in manipulating the nanostructures has been discussed. Nanoparticles of α-Fe₂O₃ (diameter ca. 15 to 130 nm) were immobilized on the cellulose fiber surface by a single step in situ hydrothermal method. However, nanoflakes of α-FeOOH having thickness ca. ~25 nm and length ca. ~250 nm were obtained by the same method in absence of cellulose fibers. A possible nucleation and growth mechanism of the formation of nanostructures on cellulose fibers have been proposed. The covalent bond formation between the cellulose fibers and nanostructures has been discussed with supporting evidence from the spectroscopic and other analytical studies such as Fourier transform infrared spectroscopy and X-ray photoelectron spectroscopy. The role of cellulose in manipulating the nanostructures has been discussed.

Keywords: cellulose fibers, α-Fe₂O₃, α-FeOOH, hydrothermal, nanoflakes, nanoparticles

Procedia PDF Downloads 150

Authors: Niklas Ravn-Boess, Nainita Bhowmick, Takamitsu Hattori, Shohei Koide, Christopher Park, Dimitris Placantonakis

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Background: Glioblastoma (GBM) is the most common and deadly primary brain malignancy in adults. Tumor propagation, brain invasion, and resistance to therapy critically depend on GBM stem-like cells (GSCs); however, the mechanisms that regulate GSC self-renewal are incompletely understood. Given the aggressiveness and poor prognosis of GBM, it is imperative to find biomarkers that could also translate into novel drug targets. Along these lines, we have identified a cell surface antigen, CD97 (ADGRE5), an adhesion G protein-coupled receptor (GPCR), that is expressed on GBM cells but is absent from non-neoplastic brain tissue. CD97 has been shown to promote invasiveness, angiogenesis, and migration in several human cancers, but its frequency of expression and functional role in regulating GBM growth and survival, and its potential as a therapeutic target has not been investigated. Design: We assessed CD97 mRNA and protein expression in patient derived GBM samples and cell lines using publicly available RNA-sequencing datasets and flow cytometry, respectively. To assess CD97 function, we generated shRNA lentiviral constructs that target a sequence in the CD97 extracellular domain (ECD). A scrambled shRNA (scr) with no predicted targets in the genome was used as a control. We evaluated CD97 shRNA lentivirally transduced GBM cells for Ki67, Annexin V, and DAPI. We also tested CD97 KD cells for their ability to self-renew using clonogenic tumorsphere formation assays. Further, we utilized synthetic Abs (sAbs) generated against the ECD of CD97 to test for potential antitumor effects using patient-derived GBM cell lines. Results: CD97 mRNA expression was expressed at high levels in all GBM samples available in the TCGA cohort. We found high levels of surface CD97 protein expression in 6/6 patient-derived GBM cell cultures, but not human neural stem cells. Flow cytometry confirmed downregulation of CD97 in CD97 shRNA lentivirally transduced cells. CD97 KD induced a significant reduction in cell growth in 3 independent GBM cell lines representing mesenchymal and proneural subtypes, which was accompanied by reduced (~20%) Ki67 staining and increased (~30%) apoptosis. Incubation of GBM cells with sAbs (20 ug/ ml) against the ECD of CD97 for 3 days induced GSC differentiation, as determined by the expression of GFAP and Tubulin. Using three unique GBM patient derived cultures, we found that CD97 KD attenuated the ability of GBM cells to initiate sphere formation by over 300 fold, consistent with an impairment in GSC self-renewal. Conclusion: Loss of CD97 expression in patient-derived GBM cells markedly decreases proliferation, induces cell death, and reduces tumorsphere formation. sAbs against the ECD of CD97 reduce tumorsphere formation, recapitulating the phenotype of CD97 KD, suggesting that sAbs that inhibit CD97 function exhibit anti-tumor activity. Collectively, these findings indicate that CD97 is necessary for the proliferation and survival of human GBM cells and identify CD97 as a promising therapeutically targetable vulnerability in GBM.

Keywords: adhesion GPCR, CD97, GBM stem cell, glioblastoma

Procedia PDF Downloads 137

Authors: Ankit Patil, Tushar D. Deshpande, Yogesh M. Nimdeo

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Pickering emulsions (PE) were developed in response to increased demand for organic, eco-friendly, and biocompatible products. These emulsions are usually stabilized by solid particles. In this research, we created chitosan-based sunflower oil-in-water (O/W) PE without the need for a surfactant. In our work, we employed chitosan, a biopolymer derived from chitin, as a stabilizer. This decision was influenced by chitosan's biocompatibility and biodegradability, as well as its anti-inflammatory and antibacterial capabilities. It also has other functional properties, such as antioxidant activity, a probiotic delivery mechanism, and the ability to encapsulate bioactive compounds. The purpose of this study was to govern key parameters that can be changed to obtain stable PE, such as the concentration of chitosan (0.3-0.5 wt.%), the concentration of oil (0.8-1 vol%), the pH of the emulsion (3-7) manipulated by the addition of 1M HCl/ 4M NaOH, and the amount of electrolyte (NaCl-0-300mM) added to increase or decrease ionic strength. A careful combination of these properties resulted in the production of the most stable and optimal PE. Particle size study found that emulsions with pH 6, 0.4% chitosan, and 300 mM salts were exceptionally stable, with droplet size 886 nm, PI of 0.1702, and zeta potential of 32.753.83 mV. It is fair to infer that when ionic strength rises, particle size, zeta potential, and PI value decrease. A lower PI value suggests that emulsion nanoparticles are more homogeneous. The addition of sodium chloride increases the ionic strength of the emulsion, facilitating the formation of more compact and ordered particle layers. These findings provide light on the creation of stimulus-responsive chitosan-based PE capable of encapsulating bioactive materials, functioning as antioxidants, and serving as food-grade emulsifiers.

Keywords: pickering emulsion, biocompatibility, eco-friendly, chitosan

Procedia PDF Downloads 238

Authors: Misty Attwood, Helgi Schioth

Abstract:

Transmembrane proteins are important components in many essential cell processes such as signal transduction, cell-cell signalling, transport of solutes, structural adhesion activities, and protein trafficking. Due to their involvement in diverse critical activities, transmembrane proteins are implicated in different disease pathways and hence are the focus of intense interest in understanding functional activities, their pathogenesis in disease, and their potential as pharmaceutical targets. Further, as the structure and function of proteins are correlated, investigating a group of proteins with the same tertiary structure, i.e., the same number of transmembrane regions, may give understanding about their functional roles and potential as therapeutic targets. In this in silico bioinformatics analysis, we identify and comprehensively characterize the previously unstudied group of proteins with five transmembrane-spanning regions (5TM). We classify nearly 60 5TM proteins in which 31 are members of ten families that contain two or more family members and all members are predicted to contain the 5TM architecture. Furthermore, nine singlet proteins that contain the 5TM architecture without paralogues detected in humans were also identifying, indicating the evolution of single unique proteins with the 5TM structure. Interestingly, more than half of these proteins function in localization activities through movement or tethering of cell components and more than one-third are involved in transport activities, particularly in the mitochondria. Surprisingly, no receptor activity was identified within this family in sharp contrast with other TM families. Three major 5TM families were identified and include the Tweety family, which are pore-forming subunits of the swelling-dependent volume regulated anion channel in astrocytes; the sidoreflexin family that acts as mitochondrial amino acid transporters; and the Yip1 domain family engaged in vesicle budding and intra-Golgi transport. About 30% of the proteins have enhanced expression in the brain, liver, or testis. Importantly, 60% of these proteins are identified as cancer prognostic markers, where they are associated with clinical outcomes of various tumour types, indicating further investigation into the function and expression of these proteins is important. This study provides the first comprehensive analysis of proteins with 5TM regions and provides details of the unique characteristics and application in pharmaceutical development.

Keywords: 5TM, cancer prognostic marker, drug targets, transmembrane protein

Procedia PDF Downloads 109

Authors: Franco Primo Soffietti, Diego Fernando Turello, Federico Pinto

Abstract:

Shear waves that propagate through the ground impose deformations that must be taken into account in the design and assessment of buried longitudinal structures such as tunnels, pipelines, and piles. Conventional engineering approaches for seismic evaluation often rely on a Euler-Bernoulli beam models supported by a Winkler foundation. This approach, however, falls short in capturing the distortions induced when the structure is subjected to shear waves. To overcome these limitations, in the present work an analytical solution is proposed considering a Timoshenko beam and including transverse and rotational springs. The present research proposes ground springs derived as closed-form analytical solutions of the equations of elasticity including the seismic wavelength. These proposed springs extend the applicability of previous plane-strain models. By considering variations in displacements along the longitudinal direction, the presented approach ensures the springs do not approach zero at low frequencies. This characteristic makes them suitable for assessing pseudo-static cases, which typically govern structural forces in kinematic interaction analyses. The results obtained, validated against existing literature and a 3D Finite Element model, reveal several key insights: i) the cutoff frequency significantly influences transverse and rotational springs; ii) neglecting displacement variations along the structure axis (i.e., assuming plane-strain deformation) results in unrealistically low transverse springs, particularly for wavelengths shorter than the structure length; iii) disregarding lateral displacement components in rotational springs and neglecting variations along the structure axis leads to inaccurately low spring values, misrepresenting interaction phenomena; iv) transverse springs exhibit a notable drop in resonance frequency, followed by increasing damping as frequency rises; v) rotational springs show minor frequency-dependent variations, with radiation damping occurring beyond resonance frequencies, starting from negative values. This comprehensive analysis sheds light on the complex behavior of embedded longitudinal structures when subjected to shear waves and provides valuable insights for the seismic assessment.

Keywords: shear waves, Timoshenko beams, Winkler springs, sol-structure interaction

Procedia PDF Downloads 61