Search results for: intracellular adhesion molecule-1
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 656

Search results for: intracellular adhesion molecule-1

536 Wheeled Robot Stable Braking Process under Asymmetric Traction Coefficients

Authors: Boguslaw Schreyer

Abstract:

During the wheeled robot’s braking process, the extra dynamic vertical forces act on all wheels: left, right, front or rear. Those forces are directed downward on the front wheels while directed upward on the rear wheels. In order to maximize the deceleration, therefore, minimize the braking time and braking distance, we need to calculate a correct torque distribution: the front braking torque should be increased, and rear torque should be decreased. At the same time, we need to provide better transversal stability. In a simple case of all adhesion coefficients being the same under all wheels, the torque distribution may secure the optimal (maximal) control of the robot braking process, securing the minimum braking distance and a minimum braking time. At the same time, the transversal stability is relatively good. At any time, we control the transversal acceleration. In the case of the transversal movement, we stop the braking process and re-apply braking torque after a defined period of time. If we correctly calculate the value of the torques, we may secure the traction coefficient under the front and rear wheels close to its maximum. Also, in order to provide an optimum braking control, we need to calculate the timing of the braking torque application and the timing of its release. The braking torques should be released shortly after the wheels passed a maximum traction coefficient (while a wheels’ slip increases) and applied again after the wheels pass a maximum of traction coefficient (while the slip decreases). The correct braking torque distribution secures the front and rear wheels, passing this maximum at the same time. It guarantees an optimum deceleration control, therefore, minimum braking time. In order to calculate a correct torque distribution, a control unit should receive the input signals of a rear torque value (which changes independently), the robot’s deceleration, and values of the vertical front and rear forces. In order to calculate the timing of torque application and torque release, more signals are needed: speed of the robot: angular speed, and angular deceleration of the wheels. In case of different adhesion coefficients under the left and right wheels, but the same under each pair of wheels- the same under right wheels and the same under left wheels, the Select-Low (SL) and select high (SH) methods are applied. The SL method is suggested if transversal stability is more important than braking efficiency. Often in the case of the robot, more important is braking efficiency; therefore, the SH method is applied with some control of the transversal stability. In the case that all adhesion coefficients are different under all wheels, the front-rear torque distribution is maintained as in all previous cases. However, the timing of the braking torque application and release is controlled by the rear wheels’ lowest adhesion coefficient. The Lagrange equations have been used to describe robot dynamics. Matlab has been used in order to simulate the process of wheeled robot braking, and in conclusion, the braking methods have been selected.

Keywords: wheeled robots, braking, traction coefficient, asymmetric

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535 Biological Treatment of Bacterial Biofilms from Drinking Water Distribution System in Lebanon

Authors: A. Hamieh, Z. Olama, H. Holail

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Drinking Water Distribution Systems provide opportunities for microorganisms that enter the drinking water to develop into biofilms. Antimicrobial agents, mainly chlorine, are used to disinfect drinking water, however, there are not yet standardized disinfection strategies with reliable efficacy and development of novel anti-biofilm strategies is still of major concern. In the present study the ability of Lactobacillus acidophilus and Streptomyces sp. cell free supernatants to inhibit the bacterial biofilm formation in Drinking Water Distribution System in Lebanon was investigated. Treatment with cell free supernatants of Lactobacillus acidophilus and Streptomyces sp. at 20% concentration resulted in average biofilm inhibition (52.89 and 39.66% respectively). A preliminary investigation about the mode of action of biofilm inhibition revealed that cell free supernatants showed no bacteriostatic or bactericidal activity against all the tested isolates. Pre-coating wells with supernatants revealed that Lactobacillus acidophilus cell free supernatant inhibited average biofilm formation (62.53%) by altering the adhesion of bacterial isolates to the surface, preventing the initial attachment step, which is important for biofilm production.

Keywords: biofilm, cell free supernatant, distribution system, drinking water, lactobacillus acidophilus, streptomyces sp, adhesion

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534 Mechanisms and Regulation of the Bi-directional Motility of Mitotic Kinesin Nano-motors

Authors: Larisa Gheber

Abstract:

Mitosis is an essential process by which duplicated genetic information is transmitted from mother to daughter cells. Incorrect chromosome segregation during mitosis can lead to genetic diseases, chromosome instability and cancer. This process is mediated by a dynamic microtubule-based intracellular structure, the mitotic spindle. One of the major factors that govern the mitotic spindle dynamics are the kinesin-5 biological nano motors that were believed to move unidirectionally on the microtubule filaments, using ATP hydrolysis, thus performing essential functions in mitotic spindle dynamics. Surprisingly, several reports from our and other laboratories have demonstrated that some kinesin-5 motors are bi-directional: they move in minus-end direction on the microtubules as single-molecules and can switch directionality under a number of conditions. These findings broke a twenty-five-years old dogma regarding kinesin directionality (1, 2). The mechanism of this bi-directional motility and its physiological significance remain unclear. To address this unresolved problem, we apply an interdisciplinary approach combining live cell imaging, biophysical single molecule, and structural experiments to examine the activity of these motors and their mutated variants in vivo and in vitro. Our data shows that factors such as protein phosphorylation (3, 4), motor clustering on the microtubules (5, 6) and structural elements (7, 8) regulate the bi-directional motility of kinesin motors. We also show, using Cryo-EM, that bi-directional kinesin motors obtain non-canonical microtubule binding, which is essential to their special motile properties and intracellular functions. We will discuss the implication of these findings to mechanism bi-directional motility and physiological roles in mitosis.

Keywords: mitosis, cancer, kinesin, microtubules, biochemistry, biophysics

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533 Biocompatible Beta Titanium Alloy Ti36Nb6Ta as a Suitable Material for Bone Regeneration

Authors: Vera Lukasova, Eva Filova, Jana Dankova, Vera Sovkova, Matej Daniel, Michala Rampichova

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Proper bone implants should promote fast adhesion of cells, stimulate cell differentiation and support the formation of bone tissue. Nowadays titanium is used as a biocompatible material capable of bone tissue integration. This study was focused on comparison of bioactive properties of two titanium alloys - beta titanium alloy Ti36Nb6Ta and standard medical titanium alloy Ti6A14V. The advantage of beta titanium alloy Ti36Nb6Ta is mainly that this material does not contain adverse elements like vanadium or aluminium. Titanium alloys were sterilized in ethanol, placed into 48 well plates and seeded with porcine mesenchymal stem cells. Cells were cultivated for 14 days in standard growth cultivation media with osteogenic supplements. Cell metabolic activity was quantified using MTS assay (Promega). Cell adhesion on day 1 and cell proliferation on further days were verified immunohistochemically using beta-actin monoclonal antibody and secondary antibody conjugated with AlexaFluor®488. Differentiation of cells was evaluated using alkaline phosphatase assay. Additionally, gene expression of collagen I was measured by qRT-PCR. Porcine mesenchymal stem cells adhered and spread well on beta titanium alloy Ti36Nb6Ta on day 1. During the 14 days’ time period the cells were spread confluently on the surface of the beta titanium alloy Ti36Nb6Ta. The metabolic activity of cells increased during the whole cultivation period. In comparison to standard medical titanium alloy Ti6A14V, we did not observe any differences. Moreover, the expression of collagen I gene revealed no statistical differences between both titanium alloys. Therefore, a beta titanium alloy Ti36Nb6Ta promotes cell adhesion, metabolic activity, proliferation and collagen I expression equally to standard medical titanium alloy Ti6A14V. Thus, beta titanium is a suitable material that provides sufficient biocompatible properties. This project was supported by the Czech Science Foundation: grant No. 16-14758S.

Keywords: beta titanium alloy, biocompatibility, differentiation, mesenchymal stem cells

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532 Surface Characteristics of Bacillus megaterium and Its Adsorption Behavior onto Dolomite

Authors: Mohsen Farahat, Tsuyoshi Hirajima

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Surface characteristics of Bacillus megaterium strain were investigated; zeta potential, FTIR and contact angle were measured. Surface energy components including Lifshitz-van der Waals, Hamaker constant, and acid/base components (Lewis acid/Lewis base) were calculated from the contact angle data. The results showed that the microbial cells were negatively charged over all pH regions with high values at alkaline region. A hydrophilic nature for the strain was confirmed by contact angle and free energy of adhesion between microbial cells. Adsorption affinity of the strain toward dolomite was studied at different pH values. The results showed that the cells had a high affinity to dolomite at acid pH comparing to neutral and alkaline pH. Extended DLVO theory was applied to calculate interaction energy between B. megaterium cells and dolomite particles. The adsorption results were in agreement with the results of Extended DLVO approach. Surface changes occurred on dolomite surface after the bio-treatment were monitored; contact angle decreased from 69° to 38° and the mineral’s floatability decreased from 95% to 25% after the treatment.

Keywords: Bacillus megaterium, surface modification, flotation, dolomite, adhesion energy

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531 Growth and Some Physiological Properties of Three Selected Species of Bifidobacteria in Admixture of Soy Milk and Goat Milk

Authors: Ahmed Zahran

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Bifidobacterium breve ATCC 15700, Bifidobacterium adolescents ATCC 15704 and Bifidobacterium longum ATCC 15707 were tested for their growth, acid production, bile tolerance, antibiotic resistance and adherence to columnar epithelial cells of the small intestine of goat. The growth of all studied species was determined in the MRSL medium. B.longum 15707 was the most active species in comparison with the other two species; it was also more resistant to bile acids. The adhesion of the studied species to the columnar epithelial cells was studied. All the studied species showed some degree of adhesion; however, B.longum adhered more than the other two species. This species was resistant to four types of antibiotics and was sensitive to chloramphenicol 30 µg. The activity of Bifidobacterium species in soymilk was evaluated by measuring the development of titratalle acidity. B.longum 15707 was the most active species in terms of growth and activity of soymilk. So, soymilk containing bifidobacteria could be added to goat milk to produce acceptable functional soy yogurt, using the ratio of (1:4) soy milk to goat milk. This product could be of unique health benefits, especially in the case of high cholesterol levels and replenishment of intestinal flora after antibiotic therapy.

Keywords: bifidobacteria physiological properties, soy milk, goat milk, attachment epithelial cells, columnar tissues, probiotic food

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530 Interaction of GCN5L1 with WHAMM and KIF5B Regulates Autolysosome Tubulation

Authors: Allen Seylani

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Lysosome-dependent autophagy is a nutrient-deprivation-induced evolutionarily conserved intracellular recycling program that sequestrates intracellular cargo into autophagosomes (AP), which then fuse with lysosomes to form autolysosomes (ALs) for cargo digestion. To restore free lysosomes, autophagic lysosome reformation (ALR) is initiated by extrusion of tubular structures from autolysosomes at the final stage of autophagy, in a process called lysosomal tubulation (LT). This project aimed to investigate the molecular mechanism of GCN5L1 in LT and the following lysosomal signaling. GCN5L1 belongs to the BORC multiprotein complexes and is involved in controlling lysosomal trafficking; however, the effect of GCN5L1 on lysosome tubulation remains largely unknown. Genetic ablation of GCN5L1 in the mouse primary hepatocytes showed dramatically increased autolysosomes (ALs), decreased lysosome regeneration and absence of lysosomal tubulation. This phenotype suggests the possibility of disruption in lysosome tubulation, which results in the disturbance of the overall lysosome homeostasis. The formation of tubulars from ALs requires kinesin motor protein KIF5B. Immunoprecipitation was employed and confirmed the interaction of GCN5L1 with the ARL8B-KIF5B complex, which recruited KIF5B to ALs. At the same time, GCN5L1 interacted with WHAMM, which promotes the actin nucleation factor, which brings actin cytoskeleton to ALs and initiates LT. Furthermore, impaired LT in GCN5L1 deficient hepatocytes was restored by overexpression of GCN5L1, and this rescue effect was attenuated by knockdown of KIF5B. Additionally, lysosomal mTORC1 activity was upregulated in GCN5L1-/- hepatocytes, while inhibition of mTORC1 abrogated the GCN5L1 mediated rescue of LT in knockout hepatocytes. Altogether these findings revealed a novel mechanism of ALR, in which a simultaneous interaction of GCN5L1 with KIF5B and WHAMM is required for LT and downstream mTORC1 signaling.

Keywords: autophagy, autolysosome, GCN5L1, lysosome

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529 LGR5 and Downstream Intracellular Signaling Proteins Play Critical Roles in the Cell Proliferation of Neuroblastoma, Meningioma and Pituitary Adenoma

Authors: Jin Hwan Cheong, Mina Hwang, Myung Hoon Han, Je Il Ryu, Young ha Oh, Seong Ho Koh, Wu Duck Won, Byung Jin Ha

Abstract:

Leucine-rich repeat-containing G-protein coupled receptor 5 (LGR5) has been reported to play critical roles in the proliferation of various cancer cells. However, the roles of LGR5 in brain tumors and the specific intracellular signaling proteins directly associated with it remain unknown. Expression of LGR5 was first measured in normal brain tissue, meningioma, and pituitary adenoma of humans. To identify the downstream signaling pathways of LGR5, siRNA-mediated knockdown of LGR5 was performed in SH-SY5Y neuroblastoma cells followed by proteomics analysis with 2-dimensional polyacrylamide gel electrophoresis (2D-PAGE). In addition, the expression of LGR5-associated proteins was evaluated in LGR5-inꠓhibited neuroblastoma cells and in human normal brain, meningioma, and pituitary adenoma tissue. Proteomics analysis showed 12 protein spots were significantly different in expression level (more than two-fold change) and subsequently identified by peptide mass fingerprinting. A protein association network was constructed from the 12 identified proteins altered by LGR5 knockdown. Direct and indirect interactions were identified among the 12 proteins. HSP 90-beta was one of the proteins whose expression was altered by LGR5 knockdown. Likewise, we observed decreased expression of proteins in the hnRNP subfamily following LGR5 knockdown. In addition, we have for the first time identified significantly higher hnRNP family expression in meningioma and pituitary adenoma compared to normal brain tissue. Taken together, LGR5 and its downstream sigꠓnaling play critical roles in neuroblastoma and brain tumors such as meningioma and pituitary adenoma.

Keywords: LGR5, neuroblastoma, meningioma, pituitary adenoma, hnRNP

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528 Identification of the Putative Interactome of Escherichia coli Glutaredoxin 2 by Affinity Chromatography

Authors: Eleni Poulou-Sidiropoulou, Charalampos N. Bompas, Martina Samiotaki, Alexios Vlamis-Gardikas

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The glutaredoxin (Grx) and thioredoxin (Trx) systems keep the intracellular environment reduced in almost all organisms. In Escherichia coli (E. coli), the Grx system relies on NADPH+ to reduce GSH reductase (GR), the latter reducing oxidized diglutathione to glutathione (GSH) which in turn reduces cytosolic Grxs, the electron donors for different intracellular substrates. In the Trx system, GR and GSH are replaced by Trx reductase (TrxR). Three of the Grxs of E. coli (Grx1, 2, 3) are reduced by GSH, while Grx4 is likely reduced by TrxR. Trx1 and Grx1 from E. coli may reduce ribonucleotide reductase Ia to ensure a constant supply of deoxyribonucleotides for the synthesis of DNA. The role of the other three Grxs is relatively unknown, especially for Grx2 that may amount up to 1 % of total cellular protein in the stationary phase of growth. The protein is known as a potent antioxidant, but no specific functions have been attributed to it. Herein, affinity chromatography of cellular extracts on immobilized Grx2, followed by MS analysis of the resulting eluates, was employed to identify protein ligands that could provide insights into the biological role of Grx2. Ionic, strong non-covalent, and covalent (disulfide) interactions with relevant proteins were detected. As a means of verification, the identified ligands were subjected to in silico docking with monothiol Grx2. In other experiments, protein extracts from E. coli cells lacking the gene for Grx2 (grxB) were compared to those of wild type. Taken together, the two approaches suggest that Grx2 is involved in protein synthesis, nucleotide metabolism, DNA damage repair, stress responses, and various metabolic processes. Grx2 appears as a versatile protein that may participate in a wide range of biological pathways beyond its known general antioxidant function.

Keywords: Escherichia coli, glutaredoxin 2, interactome, thiol-disulfide oxidoreductase

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527 Analyzing the Effects of Bio-fibers on the Stiffness and Strength of Adhesively Bonded Thermoplastic Bio-fiber Reinforced Composites by a Mixed Experimental-Numerical Approach

Authors: Sofie Verstraete, Stijn Debruyne, Frederik Desplentere

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Considering environmental issues, the interest to apply sustainable materials in industry increases. Specifically for composites, there is an emerging need for suitable materials and bonding techniques. As an alternative to traditional composites, short bio-fiber (cellulose-based flax) reinforced Polylactic Acid (PLA) is gaining popularity. However, these thermoplastic based composites show issues in adhesive bonding. This research focusses on analyzing the effects of the fibers near the bonding interphase. The research applies injection molded plate structures. A first important parameter concerns the fiber volume fraction, which directly affects adhesion characteristics of the surface. This parameter is varied between 0 (pure PLA) and 30%. Next to fiber volume fraction, the orientation of fibers near the bonding surface governs the adhesion characteristics of the injection molded parts. This parameter is not directly controlled in this work, but its effects are analyzed. Surface roughness also greatly determines surface wettability, thus adhesion. Therefore, this research work considers three different roughness conditions. Different mechanical treatments yield values up to 0.5 mm. In this preliminary research, only one adhesive type is considered. This is a two-part epoxy which is cured at 23 °C for 48 hours. In order to assure a dedicated parametric study, simple and reproduceable adhesive bonds are manufactured. Both single lap (substrate width 25 mm, thickness 3 mm, overlap length 10 mm) and double lap tests are considered since these are well documented and quite straightforward to conduct. These tests are conducted for the different substrate and surface conditions. Dog bone tensile testing is applied to retrieve the stiffness and strength characteristics of the substrates (with different fiber volume fractions). Numerical modelling (non-linear FEA) relates the effects of the considered parameters on the stiffness and strength of the different joints, obtained through the abovementioned tests. Ongoing work deals with developing dedicated numerical models, incorporating the different considered adhesion parameters. Although this work is the start of an extensive research project on the bonding characteristics of thermoplastic bio-fiber reinforced composites, some interesting results are already prominent. Firstly, a clear correlation between the surface roughness and the wettability of the substrates is observed. Given the adhesive type (and viscosity), it is noticed that an increase in surface energy is proportional to the surface roughness, to some extent. This becomes more pronounced when fiber volume fraction increases. Secondly, ultimate bond strength (single lap) also increases with increasing fiber volume fraction. On a macroscopic level, this confirms the positive effect of fibers near the adhesive bond line.

Keywords: adhesive bonding, bio-fiber reinforced composite, flax fibers, lap joint

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526 Adaptation Mechanisms of the Polyextremophile Natranaerobius Thermophilus to Saline-Alkaline-Hermal Environments

Authors: Qinghua Xing, Xinyi Tao, Haisheng Wang, Baisuo Zhao

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The first true anaerobic, halophilic alkali thermophile, Natranaerobius thermophilus DSM 18059T, serves as a valuable model for studying cellular adaptations to saline, alkaline and thermal extremes. To uncover the adaptive strategies employed by N. thermophilus in coping with these challenges, we conducted a comprehensive iTRAQ-based quantitative proteomic analysis under different conditions of salinity (3.5 M vs. 2.5 M Na+), pH (pH 9.6 vs. pH 8.6), and temperature (52°C vs. 42°C). The increased intracellular accumulation of glycine betaine, through both synthesis and transport, plays a critical role in N. thermophilus' adaptation to these combined stresses. Under all three stress conditions, the up-regulation of Trk family proteins responsible for K+ transport is observed. Intracellular K+ concentration rises in response to salt and pH levels. Multiple types of Na+/H+ antiporter (NhaC family, Mrp family and CPA family) and a diverse range of FOF1-ATP synthase are identified as vital components for maintaining ionic balance under different stress conditions. Importantly, proteins involved in amino acid metabolism, carbohydrate metabolism, ABC transporters, signaling and chemotaxis, as well as biological macromolecule repair and protection, exhibited significant up-regulation in response to these extreme conditions. These metabolic pathways emerge as critical factors in N. thermophilus' adaptation mechanisms under extreme environmental stress. To validate the proteomic data, ddPCR analysis confirmed changes in mRNA expression, thereby corroborating the up-regulation and down-regulation patterns of 19 co-up-regulated and 36 key proteins under saline, alkaline and thermal stresses. This research enriches our understanding of the complex regulatory systems that enable polyextremophiles to survive in combined extreme conditions.

Keywords: polyextremophiles, natranaerobius thermophilus, saline- alkaline- thermal stresses, combined extremes

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525 Methane Plasma Modified Polyvinyl Alcohol Scaffolds for Melanocytes Cultivation

Authors: B. Kodedova, E. Filova, M. Kralovic, E. Amler

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Vitiligo is the most common depigmentation disorder of the skin characterized by loss of melanocyte in the epidermis that leads to white lesions. One of the possible treatments is autologous transplantation of melanocytes. Biodegradable electrospun polymeric nanofibers provide good mechanical properties and could serve as suitable scaffold for epithelial cells cultivation and follow up transplantation. Moreover the microarchitecture of nanofibers mimics the structure of extracellular matrix and its porosity allows nutrients and waste exchange. The aim of this work was to develop biocompatible and biodegradable polymeric scaffolds suitable for autologous melanocytes transplantation. Electrospun polyvinyl alcohol (PVA) nanofibers were modified by cold methane plasma to lower their hydrofility and to achieve better adhesion, proliferation and viability of the murine melanocyte (Melan-a). Cells were seeded on the modified scaffolds and their adhesion, metabolic activity, proliferation and melanin synthesis was evaluated and compared to non-modified scaffolds. Results clearly indicate that cold methane plasma modified PVA nanofibers are suitable for melanocyte cultivation and may be future candidate for vitiligo treatment. Furthermore, the nanofibers can be functionalized with various bioactive substances, for enhancement of the melanocyte proliferation, melanogenesis or healing and regenerative processes. The project was supported by the Ministry of Education, Youth and Sports NPU I: LO1309 and by Grant Agency of Charles University (grant No. 1228214).

Keywords: melanocyte, nanofibers, polyvinyl alcohol, plasma modification

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524 Malachite Ore Treatment with Typical Ammonium Salts and Its Mechanism to Promote the Flotation Performance

Authors: Ayman M. Ibrahim, Jinpeng Cai, Peilun Shen, Dianwen Liu

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The difference in promoting sulfurization between different ammonium salts and its anion's effect on the sulfurization of the malachite surface was systematically studied. Therefore, this study takes malachite, a typical copper oxide mineral, as the research object, field emission scanning electron microscopy and energy-dispersive X-ray analysis (FESEM‒EDS), X-ray photoelectron spectroscopy (XPS), and other analytical and testing methods, as well as pure mineral flotation experiments, were carried out to examine the superiority of the ammonium salts as the sulfurizing reagent of malachite at the microscopic level. Additionally, the promoting effects of ammonium sulfate and ammonium phosphate on the malachite sulfurization of xanthate-flotation were compared systematically from the microstructure of sulfurized products, elemental composition, chemical state of characteristic elements, and hydrophobicity surface evolution. The FESEM and AFM results presented that after being pre-treated with ammonium salts, the adhesion of sulfurized products formed on the mineral surface was denser; thus, the flake radial dimension product was significantly greater. For malachite sulfurization flotation, the impact of ammonium phosphate in promoting sulfurization is weaker than ammonium sulfate. The reason may be that hydrolyzing phosphate consumes a substantial quantity of H+ in the solution, which hastens the formation of the copper-sulfur products, decreasing the adhesion stability of copper-sulfur species on the malachite surface.

Keywords: sulfurization flotation, adsorption characteristics, malachite, hydrophobicity

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523 Analysis of Cascade Control Structure in Train Dynamic Braking System

Authors: B. Moaveni, S. Morovati

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In recent years, increasing the usage of railway transportations especially in developing countries caused more attention to control systems railway vehicles. Consequently, designing and implementing the modern control systems to improve the operating performance of trains and locomotives become one of the main concerns of researches. Dynamic braking systems is an important safety system which controls the amount of braking torque generated by traction motors, to keep the adhesion coefficient between the wheel-sets and rail road in optimum bound. Adhesion force has an important role to control the braking distance and prevent the wheels from slipping during the braking process. Cascade control structure is one of the best control methods for the wide range of industrial plants in the presence of disturbances and errors. This paper presents cascade control structure based on two forward simple controllers with two feedback loops to control the slip ratio and braking torque. In this structure, the inner loop controls the angular velocity and the outer loop control the longitudinal velocity of the locomotive that its dynamic is slower than the dynamic of angular velocity. This control structure by controlling the torque of DC traction motors, tries to track the desired velocity profile to access the predefined braking distance and to control the slip ratio. Simulation results are employed to show the effectiveness of the introduced methodology in dynamic braking system.

Keywords: cascade control, dynamic braking system, DC traction motors, slip control

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522 Structural and Functional Characterization of the Transcriptional Regulator Rv1176 of Mycobacterium tuberculosis H37Rv

Authors: Vikash Yadav, Ashish Arora

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Microorganisms have self-defense mechanisms to protect themselves from toxic environments. Phenolic acid decarboxylase(pad) is responsible for the defense against toxicity caused by phenolic acids, converting them into less toxic vinyl derivatives. The transcription of the pad gene is regulated by a negative transcription factor, phenolic acid decarboxylase regulators (PadR), in a substrate-inducible manner. The PadR family members share the conserved DNA-binding features and interact with the operator DNA using a winged helix-turn-helix (wHTH) motif, which contains a three-helix motif and a β-stranded wing. The members of this family function as transcriptional regulators that are involved in various cellular survival processes, such as toxin production, detoxification, multidrug resistance, antibiotic biosynthesis, and carbon catabolism. Rv1176 of Mycobacterium tuberculosis H37Rv has been assigned to the PadR family protein that remains to be structurally and functionally uncharacterized. To reveal the structural mechanism by which Rv1176 could regulates effector-responsive transcription, several experiments were performed, including Electrophoretic Mobility Shift Assay (EMSA) for DNA protein interaction, differential scanning calorimetry (DSC) and Differential Scanning Fluorimetry (DSF) for temperature and ligand-dependent protein stability, Circular Dichroism (CD) spectroscopy for secondary structure analysis. Further, to evaluate the functional role of Rv1176, the intracellular survival of recombinant M. smegmatis was examined in murine macrophage cell line J774A.1 and different stressed conditions like oxidative, pH, and nutritive stress. All these studies demonstrated that Rv1176 could behave as a transcription regulator and its expression in recombinant M. smegmatis increases intracellular survival.

Keywords: EMSA, Mycobacterium tuberculosis, PadR family protein, transcriptional regulator

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521 Preparation and Characterization of Phosphate-Nickel-Titanium Composite Coating Obtained by Sol Gel Process for Corrosion Protection

Authors: Khalidou Ba, Abdelkrim Chahine, Mohamed Ebn Touhami

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A strong industrial interest is focused on the development of coatings for anticorrosion protection. In this context, phosphate composite materials are expanding strongly due to their chemical characteristics and their interesting physicochemical properties. Sol-gel coatings offer high homogeneity and purity that may lead to obtain coating presenting good adhesion to metal surface. The goal behind this work is to develop efficient coatings for corrosion protection of steel to extend its life. In this context, a sol gel process allowing to obtain thin film coatings on carbon steel with high resistance to corrosion has been developed. The optimization of several experimental parameters such as the hydrolysis time, the temperature, the coating technique, the molar ratio between precursors, the number of layers and the drying mode has been realized in order to obtain a coating showing the best anti-corrosion properties. The effect of these parameters on the microstructure and anticorrosion performance of the films sol gel coating has been investigated using different characterization methods (FTIR, XRD, Raman, XPS, SEM, Profilometer, Salt Spray Test, etc.). An optimized coating presenting good adhesion and very stable anticorrosion properties in salt spray test, which consists of a corrosive attack accelerated by an artificial salt spray consisting of a solution of 5% NaCl, pH neutral, under precise conditions of temperature (35 °C) and pressure has been obtained.

Keywords: sol gel, coating, corrosion, XPS

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520 Formation of Protective Aluminum-Oxide Layer on the Surface of Fe-Cr-Al Sintered-Metal-Fibers via Multi-Stage Thermal Oxidation

Authors: Loai Ben Naji, Osama M. Ibrahim, Khaled J. Al-Fadhalah

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The objective of this paper is to investigate the formation and adhesion of a protective aluminum-oxide (Al2O3, alumina) layer on the surface of Iron-Chromium-Aluminum Alloy (Fe-Cr-Al) sintered-metal-fibers. The oxide-scale layer was developed via multi-stage thermal oxidation at 930 oC for 1 hour, followed by 1 hour at 960 oC, and finally at 990 oC for 2 hours. Scanning Electron Microscope (SEM) images show that the multi-stage thermal oxidation resulted in the formation of predominantly Al2O3 platelets-like and whiskers. SEM images also reveal non-uniform oxide-scale growth on the surface of the fibers. Furthermore, peeling/spalling of the alumina protective layer occurred after minimum handling, which indicates weak adhesion forces between the protective layer and the base metal alloy.  Energy Dispersive Spectroscopy (EDS) analysis of the heat-treated Fe-Cr-Al sintered-metal-fibers confirmed the high aluminum content on the surface of the protective layer, and the low aluminum content on the exposed base metal alloy surface. In conclusion, the failure of the oxide-scale protective layer exposes the base metal alloy to further oxidation, and the fragile non-uniform oxide-scale is not suitable as a support for catalysts.

Keywords: high-temperature oxidation, iron-chromium-aluminum alloy, alumina protective layer, sintered-metal-fibers

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519 Proliferative Effect of Some Calcium Channel Blockers on the Human Embryonic Kidney Cell Line

Authors: Lukman Ahmad Jamil, Heather M. Wallace

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Introduction: Numerous epidemiological studies have shown a positive as well as negative association and no association in some cases between chronic use of calcium channel blockers and the increased risk of developing cancer. However, these associations were enmeshed with controversies in the absence of laboratory based studies to back up those claims. Aim: The aim of this study was to determine in mechanistic terms the association between the long-term administration of nifedipine and diltiazem and increased risk of developing cancer using the human embryonic kidney (HEK293) cell line. Methods: Cell counting using the Trypan blue dye exclusion and 3-4, 5-Dimethylthiazol-2-yl-2, 5-diphenyl-tetrazolium bromide (MTT) assays were used to investigate the effect of nifedipine and diltiazem on the growth pattern of HEK293 cells. Protein assay using modified Lowry method and analysis of intracellular polyamines concentration using Liquid Chromatography – Tandem Mass Spectrometry (LC-MS) were performed to ascertain the mechanism through which chronic use of nifedipine increases the risk of developing cancer. Results: Both nifedipine and diltiazem significantly increased the proliferation of HEK293 cells dose and time dependently. This proliferative effect after 24, 48 and 72-hour incubation period was observed at 0.78, 1.56 and 25 µM for nifedipine and 0.39, 1.56 and 25 µM for diltiazem, respectively. The increased proliferation of the cells was found to be statistically significantly (p<0.05). Furthermore, the increased proliferation of the cells induced by nifedipine was associated with the increase in the protein content and elevated intracellular polyamines concentration level. Conclusion: The chronic use of nifedipine is associated with increased proliferation of cells with concomitant elevation of polyamines concentration and elevated polyamine levels have been implicated in many malignant transformations and hence, these provide a possible explanation on the link between long term use of nifedipine and development of some human cancers. Further studies are needed to evaluate the cause of this association.

Keywords: cancer, nifedipine, polyamine, proliferation

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518 Angiogenesis and Blood Flow: The Role of Blood Flow in Proliferation and Migration of Endothelial Cells

Authors: Hossein Bazmara, Kaamran Raahemifar, Mostafa Sefidgar, Madjid Soltani

Abstract:

Angiogenesis is formation of new blood vessels from existing vessels. Due to flow of blood in vessels, during angiogenesis, blood flow plays an important role in regulating the angiogenesis process. Multiple mathematical models of angiogenesis have been proposed to simulate the formation of the complicated network of capillaries around a tumor. In this work, a multi-scale model of angiogenesis is developed to show the effect of blood flow on capillaries and network formation. This model spans multiple temporal and spatial scales, i.e. intracellular (molecular), cellular, and extracellular (tissue) scales. In intracellular or molecular scale, the signaling cascade of endothelial cells is obtained. Two main stages in development of a vessel are considered. In the first stage, single sprouts are extended toward the tumor. In this stage, the main regulator of endothelial cells behavior is the signals from extracellular matrix. After anastomosis and formation of closed loops, blood flow starts in the capillaries. In this stage, blood flow induced signals regulate endothelial cells behaviors. In cellular scale, growth and migration of endothelial cells is modeled with a discrete lattice Monte Carlo method called cellular Pott's model (CPM). In extracellular (tissue) scale, diffusion of tumor angiogenic factors in the extracellular matrix, formation of closed loops (anastomosis), and shear stress induced by blood flow is considered. The model is able to simulate the formation of a closed loop and its extension. The results are validated against experimental data. The results show that, without blood flow, the capillaries are not able to maintain their integrity.

Keywords: angiogenesis, endothelial cells, multi-scale model, cellular Pott's model, signaling cascade

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517 The Creation of Calcium Phosphate Coating on Nitinol Substrate

Authors: Kirill M. Dubovikov, Ekaterina S. Marchenko, Gulsharat A. Baigonakova

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NiTi alloys are widely used as implants in medicine due to their unique properties such as superelasticity, shape memory effect and biocompatibility. However, despite these properties, one of the major problems is the release of nickel after prolonged use in the human body under dynamic stress. This occurs due to oxidation and cracking of NiTi implants, which provokes nickel segregation from the matrix to the surface and release into living tissues. As we know, nickel is a toxic element and can cause cancer, allergies, etc. One of the most popular ways to solve this problem is to create a corrosion resistant coating on NiTi. There are many coatings of this type, but not all of them have good biocompatibility, which is very important for medical implants. Coatings based on calcium phosphate phases have excellent biocompatibility because Ca and P are the main constituents of the mineral part of human bone. This fact suggests that a Ca-P coating on NiTi can enhance osteogenesis and accelerate the healing process. Therefore, the aim of this study is to investigate the structure of Ca-P coating on NiTi substrate. Plasma assisted radio frequency (RF) sputtering was used to obtain this film. This method was chosen because it allows the crystallinity and morphology of the Ca-P coating to be controlled by the sputtering parameters. It allows us to obtain three different NiTi samples with Ca-P coating. XRD, AFM, SEM and EDS were used to study the composition, structure and morphology of the coating phase. Scratch tests were carried out to evaluate the adhesion of the coating to the substrate. Wettability tests were used to investigate the hydrophilicity of the different coatings and to suggest which of them had better biocompatibility. XRD showed that the coatings of all samples were hydroxyapatite, but the matrix was represented by TiNi intermetallic compounds such as B2, Ti2Ni and Ni3Ti. The SEM shows that the densest and defect-free coating has only one sample after three hours of sputtering. Wettability tests show that the sample with the densest coating has the lowest contact angle of 40.2° and the largest free surface area of 57.17 mJ/m2, which is mostly disperse. A scratch test was carried out to investigate the adhesion of the coating to the surface and it was shown that all coatings were removed by a cohesive mechanism. However, at a load of 30N, the indenter reached the substrate in two out of three samples, except for the sample with the densest coating. It was concluded that the most promising sputtering mode was the third, which consisted of three hours of deposition. This mode produced a defect-free Ca-P coating with good wettability and adhesion.

Keywords: biocompatibility, calcium phosphate coating, NiTi alloy, radio frequency sputtering.

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516 Effects of Cymbopogon citratus, Stapf (CS) or Lemon Grass Ethanol Extract on Antioxidant and Vascular Disorders Parameters in Rat

Authors: Suphaket Saenthaweesuk, Nutiya Somparn, Atcharaporn Thewmore

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The present study aims to investigate the effects of Cymbopogon citratus, Stapf (CS) or lemon grass ethanol extract on antioxidant and vascular disorders parameters in rat. The CS ethanol extract was screened for its phytochemical contents and antioxidant activity in vitro. Moreover, the extract was studied in rats to evaluate its effects in vivo. Rats were orally administered with CS at 1,000 mg/kg/day for 30 days. Phytochemical screening of CS extract indicated the presence of tannins, flavonoids and phenolic compounds. The extract contained phenolic compounds 1,400.10 ± 0.47 mg of gallic acid equivalents per gram CS extract. The free radical scavenging activity assessed by DPPH assay gave IC50 of 168.77 ± 3.32µg/mL, which is relatively lower than that of BHT with IC50 of 12.34 ± 1.14 µg/mL. In the animals, the protein expression of antioxidant enzymes, γ-glutamylcysteine ligase (γ-GCL) in liver was significantly increased. This was consistent with elevation of serum catalase (CAT) and superoxide dismutase (SOD) activities. However, Protein expression of vascular cell adhesion molecule-1 (VCAM-1), intercellular adhesion molecule (ICAM-1) and endothelial nitric oxide synthase (eNOS) in heart and aorta were not differenced from normal control. Taken together, the present study provides evidence that CCS water extract exhibits direct antioxidant properties and can induce cytoprotective enzymes in vivo.

Keywords: antioxidant, Cymbopogon citratus Stapf, VCAM-1, γ-glutamylcysteine ligase

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515 Antioxydant Properties and Gastroprotective Effect of Rosa canina Aqueous Extract against Alcohol-Induced Ulceration and Oxidative Stress in Rat Model

Authors: H. Sebai, M. A. Jabria, D. Wannes, H. Tounsi, L. Marzouki

Abstract:

We aimed in the present study to investigate the protective effects of Tunisian Rosa canina aqueous extract (RCAE) against ethanol-induced gastric ulceration and oxidative stress in a rat model. In this respect, adult male Wistar rats were used and divided into six groups of ten each: control, EtOH, EtOH plus various doses of RCAE, EtOH plus famotidine and EtOH + gallic acid. Phytochemical and biochemical analysis were performed using colorimetric methods. We found that RCAE is rich in total polyphenols, total flavonoids, and condensed tannins, and exhibited an importance in vitro antioxidant activity on 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical. In vivo, the results showed that oral administration of EtOH caused macroscopic and histological changes in gastric mucosa. These injuries are accompanied by an oxidative stress status as assessed by an increase of lipid peroxidation as well as a decrease of antioxidant enzyme activities such as superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx). Alcohol intoxication also induced intracellular mediators deregulation as assessed by an increase of hydrogen peroxide (H2O2), calcium and free iron levels in gastric mucosa. More, importantly, RCAE pretreatment reversed all macroscopic, histological and biochemical changes induced by EtOH administration. In conclusion, we suggest that RCAE has potent protective effects on acute ethanol-induced gastric ulceration related in part in part its antioxidant properties and its opposite effect on intracellular mediators. Indeed, Rosa canina can be offered as a food additive to protect against alcohol consumption-induced gastric and oxidative damage.

Keywords: alcohol, antioxidant properties, food additive, gastric ulceration, rat model, Rosa canina

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514 Nanostructured Multi-Responsive Coatings for Tuning Surface Properties

Authors: Suzanne Giasson, Alberto Guerron

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Stimuli-responsive polymer coatings can be used as functional elements in nanotechnologies, such as valves in microfluidic devices, as membranes in biomedical engineering, as substrates for the culture of biological tissues or in developing nanomaterials for targeted therapies in different diseases. However, such coatings usually suffer from major shortcomings, such as a lack of selectivity and poor environmental stability. The study will present multi-responsive hierarchical and hybrid polymer-based coatings aiming to overcome some of these limitations. Hierarchical polymer coatings, consisting of two-dimensional arrays of thermo-responsive cationic PNIPAM-based microgels and surface-functionalized with non-responsive or pH-responsive polymers, were covalently grafted to substrates to tune the surface chemistry and the elasticity of the surface independently using different stimuli. The characteristic dimensions (i.e., layer thickness) and surface properties (i.e., adhesion, friction) of the microgel coatings were assessed using the Surface Forces Apparatus. The ability to independently control the swelling and surface properties using temperature and pH as triggers were investigated for microgels in aqueous suspension and microgels immobilized on substrates. Polymer chain grafting did not impede the ability of cationic PNIPAM microgels to undergo a volume phase transition above the VPTT, either in suspension or immobilized on a substrate. Due to the presence of amino groups throughout the entirety of the microgel polymer network, the swelling behavior was also pH dependent. However, the thermo-responsive swelling was more significant than the pH-triggered one. The microgels functionalized with PEG exhibited the most promising behavior. Indeed, the thermo-triggered swelling of microgel-co-PEG did not give rise to changes in the microgel surface properties (i.e., surface potential and adhesion) within a wide range of pH values. It was possible for the immobilized microgel-co-PEG to undergo a volume transition (swelling/shrinking) with no change in adhesion, suggesting that the surface of the thermal-responsive microgels remains rather hydrophilic above the VPTT. This work confirms the possibility of tuning the swelling behavior of microgels without changing the adhesive properties. Responsive surfaces whose swelling properties can be reversibly and externally altered over space and time regardless of the surface chemistry are very innovative and will enable revolutionary advances in technologies, particularly in biomedical surface engineering and microfluidics, where advanced assembly of functional components is increasingly required.

Keywords: responsive materials, polymers, surfaces, cell culture

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513 iPSCs More Effectively Differentiate into Neurons on PLA Scaffolds with High Adhesive Properties for Primary Neuronal Cells

Authors: Azieva A. M., Yastremsky E. V., Kirillova D. A., Patsaev T. D., Sharikov R. V., Kamyshinsky R. A., Lukanina K. I., Sharikova N. A., Grigoriev T. E., Vasiliev A. L.

Abstract:

Adhesive properties of scaffolds, which predominantly depend on the chemical and structural features of their surface, play the most important role in tissue engineering. The basic requirements for such scaffolds are biocompatibility, biodegradation, high cell adhesion, which promotes cell proliferation and differentiation. In many cases, synthetic polymers scaffolds have proven advantageous because they are easy to shape, they are tough, and they have high tensile properties. The regeneration of nerve tissue still remains a big challenge for medicine, and neural stem cells provide promising therapeutic potential for cell replacement therapy. However, experiments with stem cells have their limitations, such as low level of cell viability and poor control of cell differentiation. Whereas the study of already differentiated neuronal cell culture obtained from newborn mouse brain is limited only to cell adhesion. The growth and implantation of neuronal culture requires proper scaffolds. Moreover, the polymer scaffolds implants with neuronal cells could demand specific morphology. To date, it has been proposed to use numerous synthetic polymers for these purposes, including polystyrene, polylactic acid (PLA), polyglycolic acid, and polylactide-glycolic acid. Tissue regeneration experiments demonstrated good biocompatibility of PLA scaffolds, despite the hydrophobic nature of the compound. Problem with poor wettability of the PLA scaffold surface could be overcome in several ways: the surface can be pre-treated by poly-D-lysine or polyethyleneimine peptides; roughness and hydrophilicity of PLA surface could be increased by plasma treatment, or PLA could be combined with natural fibers, such as collagen or chitosan. This work presents a study of adhesion of both induced pluripotent stem cells (iPSCs) and mouse primary neuronal cell culture on the polylactide scaffolds of various types: oriented and non-oriented fibrous nonwoven materials and sponges – with and without the effect of plasma treatment and composites with collagen and chitosan. To evaluate the effect of different types of PLA scaffolds on the neuronal differentiation of iPSCs, we assess the expression of NeuN in differentiated cells through immunostaining. iPSCs more effectively differentiate into neurons on PLA scaffolds with high adhesive properties for primary neuronal cells.

Keywords: PLA scaffold, neurons, neuronal differentiation, stem cells, polylactid

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512 Development of a Wall Climbing Robotic Ground Penetrating Radar System for Inspection of Vertical Concrete Structures

Authors: Md Omar Faruq Howlader, Tariq Pervez Sattar, Sandra Dudley

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This paper describes the design process of a 200 MHz Ground Penetrating Radar (GPR) and a battery powered concrete vertical concrete surface climbing mobile robot. The key design feature is a miniaturized 200 MHz dipole antenna using additional radiating arms and procedure records a reduction of 40% in length compared to a conventional antenna. The antenna set is mounted in front of the robot using a servo mechanism for folding and unfolding purposes. The robot’s adhesion mechanism to climb the reinforced concrete wall is based on neodymium permanent magnets arranged in a unique combination to concentrate and maximize the magnetic flux to provide sufficient adhesion force for GPR installation. The experiments demonstrated the robot’s capability of climbing reinforced concrete wall carrying the attached prototype GPR system and perform floor-to-wall transition and vice versa. The developed GPR’s performance is validated by its capability of detecting and localizing an aluminium sheet and a reinforcement bar (rebar) of 12 mm diameter buried under a test rig built of wood to mimic the concrete structure environment. The present robotic GPR system proves the concept of feasibility of undertaking inspection procedure on large concrete structures in hazardous environments that may not be accessible to human inspectors.

Keywords: climbing robot, dipole antenna, ground penetrating radar (GPR), mobile robots, robotic GPR

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511 Self-Assembled Laser-Activated Plasmonic Substrates for High-Throughput, High-Efficiency Intracellular Delivery

Authors: Marinna Madrid, Nabiha Saklayen, Marinus Huber, Nicolas Vogel, Christos Boutopoulos, Michel Meunier, Eric Mazur

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Delivering material into cells is important for a diverse range of biological applications, including gene therapy, cellular engineering and imaging. We present a plasmonic substrate for delivering membrane-impermeable material into cells at high throughput and high efficiency while maintaining cell viability. The substrate fabrication is based on an affordable and fast colloidal self-assembly process. When illuminated with a femtosecond laser, the light interacts with the electrons at the surface of the metal substrate, creating localized surface plasmons that form bubbles via energy dissipation in the surrounding medium. These bubbles come into close contact with the cell membrane to form transient pores and enable entry of membrane-impermeable material via diffusion. We use fluorescence microscopy and flow cytometry to verify delivery of membrane-impermeable material into HeLa CCL-2 cells. We show delivery efficiency and cell viability data for a range of membrane-impermeable cargo, including dyes and biologically relevant material such as siRNA. We estimate the effective pore size by determining delivery efficiency for hard fluorescent spheres with diameters ranging from 20 nm to 2 um. To provide insight to the cell poration mechanism, we relate the poration data to pump-probe measurements of micro- and nano-bubble formation on the plasmonic substrate. Finally, we investigate substrate stability and reusability by using scanning electron microscopy (SEM) to inspect for damage on the substrate after laser treatment. SEM images show no visible damage. Our findings indicate that self-assembled plasmonic substrates are an affordable tool for high-throughput, high-efficiency delivery of material into mammalian cells.

Keywords: femtosecond laser, intracellular delivery, plasmonic, self-assembly

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510 Dry Modifications of PCL/Chitosan/PCL Tissue Scaffolds

Authors: Ozan Ozkan, Hilal Turkoglu Sasmazel

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Natural polymers are widely used in tissue engineering applications, because of their biocompatibility, biodegradability and solubility in the physiological medium. On the other hand, synthetic polymers are also widely utilized in tissue engineering applications, because they carry no risk of infectious diseases and do not cause immune system reaction. However, the disadvantages of both polymer types block their individual usages as tissue scaffolds efficiently. Therefore, the idea of usage of natural and synthetic polymers together as a single 3D hybrid scaffold which has the advantages of both and the disadvantages of none has been entered to the literature. On the other hand, even though these hybrid structures support the cell adhesion and/or proliferation, various surface modification techniques applied to the surfaces of them to create topographical changes on the surfaces and to obtain reactive functional groups required for the immobilization of biomolecules, especially on the surfaces of synthetic polymers in order to improve cell adhesion and proliferation. In a study presented here, to improve the surface functionality and topography of the layer by layer electrospun 3D poly-epsilon-caprolactone/chitosan/poly-epsilon-caprolactone hybrid tissue scaffolds by using atmospheric pressure plasma method, thus to improve cell adhesion and proliferation of these tissue scaffolds were aimed. The formation/creation of the functional hydroxyl and amine groups and topographical changes on the surfaces of scaffolds were realized by using two different atmospheric pressure plasma systems (nozzle type and dielectric barrier discharge (DBD) type) carried out under different gas medium (air, Ar+O2, Ar+N2). The plasma modification time and distance for the nozzle type plasma system as well as the plasma modification time and the gas flow rate for DBD type plasma system were optimized with monitoring the changes in surface hydrophilicity by using contact angle measurements. The topographical and chemical characterizations of these modified biomaterials’ surfaces were carried out with SEM and ESCA, respectively. The results showed that the atmospheric pressure plasma modifications carried out with both nozzle type plasma and DBD plasma caused topographical and functionality changes on the surfaces of the layer by layer electrospun tissue scaffolds. However, the shelf life studies indicated that the hydrophilicity introduced to the surfaces was mainly because of the functionality changes. Therefore, according to the optimized results, samples treated with nozzle type air plasma modification applied for 9 minutes from a distance of 17 cm and Ar+O2 DBD plasma modification applied for 1 minute under 70 cm3/min O2 flow rate were found to have the highest hydrophilicity compared to pristine samples.

Keywords: biomaterial, chitosan, hybrid, plasma

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509 Reasons for Adhesion of Membership: A Case Study of Brazilian Soccer Team

Authors: Alexandre Olkoski, Marcelo Curth

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Football in Brazil is considered a passion, being the most popular sport in the country, both by the consumer public and by the means of communication that divulge it individually, when compared with other sports modalities. In the last two decades, the soccer teams have given greater importance to the management, since they understood that the same should be managed as a company, but with peculiarities related to the business. In this sense, Brazilian soccer clubs started to make bigger investments for the adhesion of fans in their social frames, allowing a greater need of understanding about the profile of this group of fans/clients. Thus, this work aims to understand the reasons that cause the fans to join the club and identify variables present in the process of intention to join the club. For that, a qualitative exploratory research was conducted, in which thirty-one membership of a soccer club from southern Brazil were interviewed. Based on the interviews, five categories were classified as emotional aspects (passion and love), cognitive aspects (easy access to the stadium and promotional values in tickets), external influences (family and friends), situational aspects (club moment) and aspects related to the event (engagement by modality). As results found in the analysis, it can be highlighted that the motivation of the majority of the respondents to become a member of the analyzed club, is related to the emotional aspects, such as passion and love. Thus, it is perceived that sport, in the case of soccer, generates in the involved ones (fans and leaders) different manifestations, suggesting that the management of this type of business has great complexity and should not be observed only by the spectrum of the club like a business.

Keywords: consumer behavior, marketing, membership, soccer

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508 hsa-miR-1204 and hsa-miR-639 Prominent Role in Tamoxifen's Molecular Mechanisms on the EMT Phenomenon in Breast Cancer Patients

Authors: Mahsa Taghavi

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In the treatment of breast cancer, tamoxifen is a regularly prescribed medication. The effect of tamoxifen on breast cancer patients' EMT pathways was studied. In this study to see if it had any effect on the cancer cells' resistance to tamoxifen and to look for specific miRNAs associated with EMT. In this work, we used continuous and integrated bioinformatics analysis to choose the optimal GEO datasets. Once we had sorted the gene expression profile, we looked at the mechanism of signaling, the ontology of genes, and the protein interaction of each gene. In the end, we used the GEPIA database to confirm the candidate genes. after that, I investigated critical miRNAs related to candidate genes. There were two gene expression profiles that were categorized into two distinct groups. Using the expression profile of genes that were lowered in the EMT pathway, the first group was examined. The second group represented the polar opposite of the first. A total of 253 genes from the first group and 302 genes from the second group were found to be common. Several genes in the first category were linked to cell death, focal adhesion, and cellular aging. Two genes in the second group were linked to cell death, focal adhesion, and cellular aging. distinct cell cycle stages were observed. Finally, proteins such as MYLK, SOCS3, and STAT5B from the first group and BIRC5, PLK1, and RAPGAP1 from the second group were selected as potential candidates linked to tamoxifen's influence on the EMT pathway. hsa-miR-1204 and hsa-miR-639 have a very close relationship with the candidates genes according to the node degrees and betweenness index. With this, the action of tamoxifen on the EMT pathway was better understood. It's important to learn more about how tamoxifen's target genes and proteins work so that we can better understand the drug.

Keywords: tamoxifen, breast cancer, bioinformatics analysis, EMT, miRNAs

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507 Wear Resistance and Thermal Stability of Tungsten Boride Layers Deposited by Magnetron Sputtering

Authors: Justyna Chrzanowska, Jacek Hoffman, Dariusz Garbiec, Łukasz Kurpaska, Piotr Denis, Tomasz Moscicki, Zygmunt Szymanski

Abstract:

Tungsten and boron compounds belong to the group of superhard materials and its hardness could exceed 40 GPa. In this study, the properties of the tungsten boride (WB) layers deposited in magnetron sputtering process are investigated. The sputtering process occurred from specially prepared targets that were composed of boron and tungsten mixed in molar ratio of 2.5 or 4.5 and sintered in spark plasma sintering process. WB layers were deposited on silicon (100) and stainless steel 304 substrates at room temperature (RT) or in 570 °C. Layers deposited in RT and in elevated temperature varied considerably. Layers deposited in RT are amorphous and have low adhesion. In contrast, the layers deposited in 570 °C are crystalline and have good adhesion. All deposited layers have a hardness about 40 GPa. Moreover, the friction coefficient of crystalline layers is 0.22 and wear rate is about 0.67•10-6 mm3N-1m-1. After material characterization the WB layers were annealed in argon atmosphere in 1000 °C for 1 hour. On the basis of X-Ray Diffraction analysis, it has been noted that the crystalline layers are thermally stable and do not change their phase composition, whereas the amorphous layers change their phase composition. Moreover, after annealing, on the surface of WB layers some cracks were observed. It is probably connected with the differences of the thermal expansion between the layer and the substrate. Despite of the presence of cracks, the wear resistance of annealed layers is still higher than the wear resistance of uncoated substrate. The analysis of the structure and properties of tungsten boride layers lead to the discussion about the application area of this material.

Keywords: hard coatings, hard materials, magnetron sputtering, mechanical properties, tungsten boride

Procedia PDF Downloads 252