Search results for: hela cell line

Authors: Paua Ketan, Bhagate Rajkumar, Adiga Ganesh, M. Kiran

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Low gear noise is an important sound quality feature in modern passenger cars. Annoying gear noise from the gearbox is influenced by the gear design, gearbox shaft layout, manufacturing deviations in the components, assembly errors and the mounting arrangement of the complete gearbox. Geometrical deviations in the form of profile and lead errors are often present on the flanks of the inspected gears. Ghost frequencies of a gear are very challenging to identify in standard gear measurement and analysis process due to small wavelengths involved. In this paper, gear whine noise occurring at non-integral multiples of gear mesh frequency of passenger car gearbox is investigated and the root cause is identified using the displacement deviation analysis (DDA) method. DDA method is applied to identify ghost frequency excitations on the flanks of gears arising out of generation grinding. Frequency identified through DDA correlated with the frequency of vibration and noise on the end-of-line machine as well as vehicle level measurements. With the application of DDA method along with standard lead profile measurement, gears with ghost frequency geometry deviations were identified on the production line to eliminate defective parts and thereby eliminate ghost frequency noise from a vehicle. Further, displacement deviation analysis can be used in conjunction with the manufacturing process simulation to arrive at suitable countermeasures for arresting the ghost frequency.

Keywords: displacement deviation analysis, gear whine, ghost frequency, sound quality

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Authors: Okyu Kwon, Dongho Kang, Byungsik Kim, Seungkwon Jung

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We propose a technique to measure the impact of loss of traffic function in a particular area to surrounding areas. The proposed method is applied to the city of Seoul, which is the capital of South Korea, with a population of about ten million. Based on the actual road network in Seoul, we construct an abstract road network between 1kmx1km grid cells. The link weight of the abstract road network is re-adjusted considering traffic volume measured at several survey points. On the modified abstract road network, we evaluate the traffic vulnerability by calculating a network measure of betweenness centrality (BC) for every single grid cells. This study analyzes traffic impacts caused by road dysfunction due to heavy rainfall in urban areas. We could see the change of the BC value in all other grid cells by calculating the BC value once again when the specific grid cell lost its traffic function, that is, when the node disappeared on the grid-based road network. The results show that it is appropriate to use the sum of the BC variation of other cells as the influence index of each lattice cell on traffic. This research was supported by a grant (2017-MOIS31-004) from Fundamental Technology Development Program for Extreme Disaster Response funded by Korean Ministry of Interior and Safety (MOIS).

Keywords: vulnerability, road network, beweenness centrality, heavy rainfall, road impact

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Authors: Khobaib Khobaib, Alexander Mikkelsen, Zbigniew Rozynek

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Electric fields have proven useful for inducing droplet deformation and to structure particles adsorbed at droplet interfaces. In this experimental research, direct current electric fields were applied to deform particle-covered droplets made out of silicone oil and immersed in castor oil. The viscosity of the drop and surrounding fluid were changed by external heating. We designed an experimental system in such a way that electric field-induced electrohydrodynamic (EHD) flows were asymmetric and only present on one side of the drop, i.e., the droplet adjoined a washer and adhered to one of the electrodes constituting the sample cell. The study investigated the influence of viscosity on the steady-state deformation magnitude of particle-laden droplets, droplet compression, and relaxation, as well as particle arrangements at drop interfaces. Initially, before the application of an electric field, we changed the viscosity of the fluids by heating the sample cell at different temperatures. The viscosity of the fluids was varied by changing the temperature of the fluids from 25 to 50°C. Under the application of a uniform electric field of strength 290 Vmm⁻¹, electric stress was induced at the drop interface, yielding drop deformation. In our study, we found that by lowering the fluid viscosity, the velocity of the EHD flows was increased, which also increases the deformation of the drop.

Keywords: drop deformation and relaxation, electric field, electrohydrodynamic flow, particle assembly, viscosity

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Authors: Ádám Vass, István Bakos, Irina Borbáth, Zoltán Pászti, István Sajó, András Tompos

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Important requirements for the anode side electrocatalysts of polymer electrolyte membrane (PEM) fuel cells are CO-tolerance, stability and corrosion resistance. Carbon is still the most common material for electrocatalyst supports due to its low cost, high electrical conductivity and high surface area, which can ensure good dispersion of the Pt. However, carbon becomes degraded at higher potentials and it causes problem during application. Therefore it is important to explore alternative materials with improved stability. Molybdenum-oxide can improve the CO-tolerance of the Pt/C catalysts, but it is prone to leach in acidic electrolyte. The Mo was stabilized by isovalent substitution of molybdenum into the rutile phase titanium-dioxide lattice, achieved by a modified multistep sol-gel synthesis method optimized for preparation of Ti0.7Mo.3O2-C composite. High degree of Mo incorporation into the rutile lattice was developed. The conductivity and corrosion resistance across the anticipated potential/pH window was ensured by mixed oxide – activated carbon composite. Platinum loading was carried out using NaBH4 and ethylene glycol; platinum content was 40 wt%. The electrocatalyst was characterized by both material investigating methods (i.e. XRD, TEM, EDS, XPS techniques) and electrochemical methods (cyclic-voltammetry, COads stripping voltammetry, hydrogen oxidation reaction on rotating disc electrode). The electrochemical activity of the sample was compared to commercial 40 wt% Pt/C (Quintech) and PtRu/C (Quintech, Pt= 20 wt%, Ru= 10 wt%) references. Enhanced CO tolerance of the electrocatalyst prepared using the Ti0.7Mo.3O2-C composite material was evidenced by the appearance of a CO-oxidation related 'pre-peak' and by the pronounced shift of the maximum of the main CO oxidation peak towards less positive potential compared to Pt/C. Fuel cell polarization measurements were also carried out using Bio-Logic and Paxitech FCT-150S test device. All details on the design, preparation, characterization and testing by both electrochemical measurements and fuel cell test device of electrocatalyst supported on Ti0.7Mo.3O2-C composite material will be presented and discussed.

Keywords: anode electrocatalyst, composite material, CO-tolerance, TiMoOx

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Authors: Ramesh P., Aby Joseph

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Micro Inverters provide Module Embedded Solution for harvesting energy from small-scale solar photovoltaic (PV) panels. In addition to higher modularity & reliability (25 years of life), the MicroInverter has inherent advantages such as avoidance of long DC cables, eliminates module mismatch losses, minimizes partial shading effect, improves safety and flexibility in installations etc. Due to the above-stated benefits, the renewable energy technology with Solar Photovoltaic (PV) Micro Inverter becomes more widespread in Village Nano Grid application ensuring grid independence for rural communities and areas without access to electricity. While the primary objective of this paper is to discuss the problems related to rural electrification, this concept can also be extended to urban installation with grid connectivity. This work presents a comprehensive analysis of the power circuit design, control methodologies and prototyping of 300Wₚ Single Phase PV Micro Inverter. This paper investigates two different topologies for PV Micro Inverters, based on the first hand on Single Stage Flyback/ Forward PV Micro-Inverter configuration and the other hand on the Double stage configuration including DC-DC converter, H bridge DC-AC Inverter. This work covers Power Decoupling techniques to reduce the input filter capacitor size to buffer double line (100 Hz) ripple energy and eliminates the use of electrolytic capacitors. The propagation of the double line oscillation reflected back to PV module will affect the Maximum Power Point Tracking (MPPT) performance. Also, the grid current will be distorted. To mitigate this issue, an independent MPPT control algorithm is developed in this work to reject the propagation of this double line ripple oscillation to PV side to improve the MPPT performance and grid side to improve current quality. Here, the power hardware topology accepts wide input voltage variation and consists of suitably rated MOSFET switches, Galvanically Isolated gate drivers, high-frequency magnetics and Film capacitors with a long lifespan. The digital controller hardware platform inbuilt with the external peripheral interface is developed using floating point microcontroller TMS320F2806x from Texas Instruments. The firmware governing the operation of the PV Micro Inverter is written in C language and was developed using code composer studio Integrated Development Environment (IDE). In this work, the prototype hardware for the Single Phase Photovoltaic Micro Inverter with Double stage configuration was developed and the comparative analysis between the above mentioned configurations with experimental results will be presented.

Keywords: double line oscillation, micro inverter, MPPT, nano grid, power decoupling

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Authors: Elsadig Naseraddeen Ahmed Mohamed

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In contrast to the uncertainty and complementary principle, it will be shown in the present paper that the probability of the simultaneous occupation event of any definite values of coordinates by any definite values of momentum and energy at any definite instance of time can be described by a binary definite function equivalent to the difference between their numbers of occupation and evacuation epochs up to that time and also equivalent to the number of exchanges between those occupation and evacuation epochs up to that times modulus two, these binary definite quantities can be defined at all point in the time’s real-line so it form a binary signal represent a complete mechanical description of physical reality, the time of these exchanges represent the boundary of occupation and evacuation epochs from which we can calculate these binary signals using the fact that the time of universe events actually extends in the positive and negative of time’s real-line in one direction of extension when these number of exchanges increase, so there exists noninvertible transformation matrix can be defined as the matrix multiplication of invertible rotation matrix and noninvertible scaling matrix change the direction and magnitude of exchange event vector respectively, these noninvertible transformation will be called actual transformation in contrast to information transformations by which we can navigate the universe’s events transformed by actual transformations backward and forward in time’s real-line, so these information transformations will be derived as an elements of a group can be associated to their corresponded actual transformations. The actual and information model of the universe will be derived by assuming the existence of time instance zero before and at which there is no coordinate occupied by any definite values of momentum and energy, and then after that time, the universe begin its expanding in spacetime, this assumption makes the need for the existence of Laplace’s demon who at one moment can measure the positions and momentums of all constituent particle of the universe and then use the law of classical mechanics to predict all future and past of universe’s events, superfluous, we only need for the establishment of our analog to digital converters to sense the binary signals that determine the boundaries of occupation and evacuation epochs of the definite values of coordinates relative to its origin by the definite values of momentum and energy as present events of the universe from them we can predict approximately in high precision it's past and future events.

Keywords: binary-signal mechanics, actual-information model of the universe, actual-transformation, information-transformation, uncertainty principle, Laplace's demon

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Authors: Taknaz Banihashemi, Jiangang Fei, Peggy Shu-Ling Chen

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Reverse logistics (RL) is a part of the logistics of companies and its aim is to reclaim value from the returned products in an environmentally friendly manner. In recent years, RL has attracted significant attention among both practitioners and academics due to environmental directives and governmental legislation, consumer concerns and social responsibilities for environment, awareness of the limits of natural resources and economic potential. Sustainability development is considered as a critical goal for organisations due to its impact on competitive advantage. With growing environmental concerns and legal regulations related to green and sustainability issues, product disposition through RL can be considered as an environmental, economic and social sound way to achieve sustainable development. When employed properly, RL can help firms to improve their sustainability performance. The aim of this paper is to investigate the sustainability issues in the context of RL in the perspective of the triple-bottom-line approach. Content analysis was used to collect the information. The findings show that there is a research gap to investigate the relationship between RL and sustainability performance. Most of the studies have focused on performance evaluation of RL by considering the factors related to economic and environmental performance. RL can have significant effects on social issues along with economic and environmental issues. The inclusion of the social aspect in the sustainability performance will provide a complete and holistic picture of how RL may impact on the sustainability performance of firms. Generally, there is a lack of research on investigating the relationship between RL and sustainability by integrating the three pillars of triple-bottom-line sustainability performance. This paper provides academics and researchers a broad view of the correlations between RL and sustainability performance.

Keywords: verse Logistics, review, sustainability, sustainability performance

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Authors: Robson da Cunha Santos, Caio Cezar R. Bonifacio, Diego Mureb Quesada, Gerson Gomes Cunha

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The paper is related to the safety of oil wells and environmental preservation on the planet, because they require great attention and commitment from oil companies and people who work with these equipments. This must occur from drilling the well until it is abandoned in order to safeguard the environment and prevent possible damage. The project had as main objective the constitution resulting from comparatives made among books, articles and publications with information gathered in technical visits to operational bases of Petrobras. After the visits, the information from methods of utilization and present managements, which were not available before, became available to the general audience. As a result, it is observed a huge flux of incorrect and out-of-date information that comprehends not only bibliographic archives, but also academic resources and materials. During the gathering of more in-depth information on the manufacturing, assembling, and use aspects of DHSVs, several issues that were previously known as correct, customary issues were discovered to be uncertain and outdated. Information of great importance resulted in affirmations about subjects as the depth of the valve installation that was before installed to 30 meters from the seabed (mud line). Despite this, the installation should vary in conformity to the ideal depth to escape from area with the biggest tendency to hydrates formation according to the temperature and pressure. Regarding to valves with nitrogen chamber, in accordance with books, they have their utilization linked to water line ≥ 700 meters, but in Brazilian exploratory fields, their use occurs from 600 meters of water line. The valves used in Brazilian fields are able to be inserted to the production column and self-equalizing, but the use of screwed valve in the column of production and equalizing is predominant. Although these valves are more expensive to acquire, they are more reliable, efficient, with a bigger shelf life and they do not cause restriction to the fluid flux. It follows that based on researches and theoretical information confronted to usual forms used in fields, the present project is important and relevant. This project will be used as source of actualization and information equalization that connects academic environment and real situations in exploratory situations and also taking into consideration the enrichment of precise and easy to understand information to future researches and academic upgrading.

Keywords: down hole safety valve, security devices, installation, oil-wells

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Authors: Yanni Chang, Dezhi Dai, Albert Y. Tong

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Piecewise linear interface calculation (PLIC) schemes are widely used in the volume-of-fluid (VOF) method to capture interfaces in numerical simulations of multiphase flows. Dynamic overset meshes can be especially useful in applications involving component motions and complex geometric shapes. In the present study, the VOF value of an acceptor cell is evaluated in a geometric way that transfers the fraction field between the meshes precisely with reconstructed interfaces from the corresponding donor elements. The acceptor cell value is evaluated by using a weighted average of its donors for most of the overset interpolation schemes for continuous flow variables. The weighting factors are obtained by different algebraic methods. Unlike the continuous flow variables, the VOF equation is a step function near the interfaces, which ranges from zero to unity rapidly. A geometric interpolation scheme of the VOF field in overset meshes for the PLIC-VOF method has been proposed in the paper. It has been tested successfully in quadrilateral/hexahedral overset meshes by employing several VOF advection tests with imposed solenoidal velocity fields. The proposed algorithm has been shown to yield higher accuracy in mass conservation and interface reconstruction compared with three other algebraic ones.

Keywords: interpolation scheme, multiphase flows, overset meshes, PLIC-VOF method

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Authors: F. Kashanian, M. M. Masoudi, A. Akbari, A. Shamloo, M. R. Zand, S. S. Salehi

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Nano-sized materials present new opportunities in biology and medicine and they are used as biomedical tools for investigation, separation of molecules and cells. To achieve more effective cancer therapy, it is essential to select cancer cells exactly. This research suggests that using the antibody-functionalized nontoxic Arginine-doped magnetic nanoparticles (A-MNPs), has been prosperous in detection, capture, and magnetic separation of circulating tumor cells (CTCs) in tumor tissue. In this study, A-MNPs were synthesized via a simple precipitation reaction and directly immobilized Ep-CAM EBA-1 antibodies over superparamagnetic A-MNPs for Mucin BCA-225 in breast cancer cell. The samples were characterized by vibrating sample magnetometer (VSM), FT-IR spectroscopy, Tunneling Electron Microscopy (TEM) and Scanning Electron Microscopy (SEM). These antibody-functionalized nontoxic A-MNPs were used to capture breast cancer cell. Through employing a strong permanent magnet, the magnetic separation was achieved within a few seconds. Antibody-Conjugated nontoxic Arginine-doped Fe₃O₄ nanoparticles have the potential for the future study to capture CTCs which are released from tumor tissue and for drug delivery, and these results demonstrate that the antibody-conjugated A-MNPs can be used in magnetic hyperthermia techniques for cancer treatment.

Keywords: tumor tissue, antibody, magnetic nanoparticle, CTCs capturing

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Authors: Rafael Martins Manzano Silva, Jean-Francois Tremong

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Electrification of railway lines allows to increase speed, power, capacity and energetic efficiency of rolling stocks. However, this process of electrification is complex and costly. An electrification project is not just about design of catenary. It also includes installation of structures around electrification, as substation installation, electrical isolation, signalling, telecommunication and civil engineering structures. France has more than 30,000 km of railways, whose only 53% are electrified. The others 47% of railways use diesel locomotive and represent only 10% of the circulation (tons.km). For this reason, a new type of electrification, less expensive than the usual, is requested to enable the modernization of these railways. One solution could be the use of hybrids trains. This technology opens up new opportunities for less expensive infrastructure development such as the partial electrification of railway lines. In a partially electrified railway, the power supply of theses hybrid trains could be made either by the catenary or by the on-board energy storage system (ESS). Thus, the on-board ESS would feed the energetic needs of the train along the non-electrified zones while in electrified zones, the catenary would feed the train and recharge the on-board ESS. This paper’s objective deals with the technical and economic potential identification of partial electrification of railway lines. This study provides different scenarios of electrification by replacing the most expensive places to electrify using on-board ESS. The target is to reduce the cost of new electrification projects, i.e. reduce the cost of electrification infrastructures while not increasing the cost of rolling stocks. In this study, scenarios are constructed in function of the electrification’s cost of each structure. The electrification’s cost varies considerably because of the installation of catenary support in tunnels, bridges and viaducts is much more expensive than in others zones of the railway. These scenarios will be used to describe the power supply system and to choose between the catenary and the on-board energy storage depending on the position of the train on the railway. To identify the influence of each partial electrification scenario in the sizing of the on-board ESS, a model of the railway line and of the rolling stock is developed for a real case. This real case concerns a railway line located in the south of France. The energy consumption and the power demanded at each point of the line for each power supply (catenary or on-board ESS) are provided at the end of the simulation. Finally, the cost of a partial electrification is obtained by adding the civil engineering costs of the zones to be electrified plus the cost of the on-board ESS. The study of the technical and economic potential ends with the identification of the most economically interesting scenario of electrification.

Keywords: electrification, hybrid, railway, storage

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Authors: Fadia Gafri, Kathryn Mckintosh, Louise Young, Alan Harvey, Simon Mackay, Andrew Paul, Robin Plevin

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Nuclear factor-kappa B (NF-κB) is a ubiquitous transcription factor found originally to play a key role in regulating inflammation. However considerable evidence links this pathway to the suppression of apoptosis, cellular transformation, proliferation and invasion (Aggarwal et al., 2006). Moreover, recent studies have also linked inflammation to cancer progression making NF-κB overall a promising therapeutic target for drug discovery (Dobrovolskaia & Kozlov, 2005). In this study we examined the effect of the natural product canthin-6-one (SU182) as part of a CRUK small molecule drug discovery programme for effects upon the NF-κB pathway. Initial studies demonstrated that SU182 was found to have good potency against the inhibitory kappa B kinases (IKKs) at 30M in vitro. However, at concentrations up to 30M, SU182 had no effect upon TNFα stimulated loss in cellular IκBα or p65 phosphorylation in the keratinocyte cell line NCTC2544. Nevertheless, 30M SU182 reduced TNF-α / PMA-induced NF-κB-linked luciferase reporter activity to (22.9 ± 5%) and (34.6± 3 %, P<0.001) respectively, suggesting an action downstream of IKK signalling. Indeed, SU182 neither decreased NF-κB-DNA binding as assayed by EMSA nor prevented the translocation of p65 (NF-κB) to the nucleus assessed by immunofluorescence and subcellular fractionation. In addition to the inhibition of transcriptional activity of TNFα-induced NF-κB reporter activity SU182 significantly reduced PMA-induced AP-1-linked luciferase reporter activity to about (48± 9% at 30M, P<0.001) . This mode of inhibition was not sufficient to prevent the activation of NF-κB dependent induction of other proteins such as COX-2 and iNOS, or activated MAP kinases (p38, JNK and ERK1/2) in LPS stimulated RAW 264.7 macrophages. Taken together these data indicate the potential for SU182 to interfere with the transcription factors NF-κB and AP-1 at transcriptional level. However, no potential anti-inflammatory effect was indicated, further investigation for other NF-κB dependent proteins linked to survival are also required to identify the exact mechanism of action.

Keywords: Canthin-6-one, NF-κB, AP-1, phosphorylation, Nuclear translocation, DNA-binding activity, inflammatory proteins.

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Authors: Ali Riza Perçin, Eser Bingül

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Internet technologies have created opportunities with which people share their ideologies, thoughts and products. This virtual world, named social media has given the chance of gathering individual users and people from the world's remote locations and establishing an interaction between them. However, to an increasingly higher degree terrorist organizations today use the internet and most notably social-network media to create the effects they desire through a series of on-line activities. These activities, designed to support their activities, include information collection (intelligence), target selection, propaganda, fundraising and recruitment to name a few. Meanwhile, these have been used as the most important tool for recruitment especially from the different region of the world, especially disenfranchised youth, in the West in order to mobilize support and recruit “foreign fighters.” The recruits have obtained the statue, which is not accessible in their society and have preferred the style of life that is offered by the terrorist organizations instead of their current life. Like other terrorist groups, for a while now the terrorist organization Islamic State (IS) in Iraq and Syria has employed a social-media strategy in order to advance their strategic objectives. At the moment, however, IS seems to be more successful in their on-line activities than other similar organizations. IS uses social media strategically as part of its armed activities and for the sustainability of their military presence in Syria and Iraq. In this context, “Generation Y”, which could exist at the critical position and undertake active role, has been examined. Additionally, the explained characteristics of “Generation Y” have been put forward and the duties of families and society have been stated as well.

Keywords: social media, "generation Y", terrorist organization, islamic state IS

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Authors: Takashi Mitsuishi

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Fuzzy logic has gained acceptance in the recent years in the fields of social sciences and humanities such as psychology and linguistics because it can manage the fuzziness of words and human subjectivity in a logical manner. However, the major field of application of the fuzzy logic is control engineering as it is a part of the set theory and mathematical logic. Mamdani method, which is the most popular technique for approximate reasoning in the field of fuzzy control, is one of the ways to numerically represent the control afforded by human language and sensitivity and has been applied in various practical control plants. Fuzzy logic has been gradually developing as an artificial intelligence in different applications such as neural networks, expert systems, and operations research. The objects of inference vary for different application fields. Some of these include time, angle, color, symptom and medical condition whose fuzzy membership function is a periodic function. In the defuzzification stage, the domain of the membership function should be unique to obtain uniqueness its defuzzified value. However, if the domain of the periodic membership function is determined as unique, an unintuitive defuzzified value may be obtained as the inference result using the center of gravity method. Therefore, the authors propose a method of circular-polar-coordinates transformation and defuzzification of the periodic membership functions in this study. The transformation to circular polar coordinates simplifies the domain of the periodic membership function. Defuzzified value in circular polar coordinates is an argument. Furthermore, it is required that the argument is calculated from a closed plane figure which is a periodic membership function on the circular polar coordinates. If the closed plane figure is continuous with the continuity of the membership function, a significant amount of computation is required. Therefore, to simplify the practice example and significantly reduce the computational complexity, we have discretized the continuous interval and the membership function in this study. In this study, the following three methods are proposed to decide the argument from the discrete polygon which the continuous plane figure is transformed into. The first method provides an argument of a straight line passing through the origin and through the coordinate of the arithmetic mean of each coordinate of the polygon (physical center of gravity). The second one provides an argument of a straight line passing through the origin and the coordinate of the geometric center of gravity of the polygon. The third one provides an argument of a straight line passing through the origin bisecting the perimeter of the polygon (or the closed continuous plane figure).

Keywords: defuzzification, fuzzy membership function, periodic function, polar coordinates transformation

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Authors: Lynn Lehmann, Dinorah Leyva, Ana Lazic, Stefan Duhr, Philipp Baaske

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Characterization of biomolecular interactions, such as protein-protein, protein-nucleic acid or protein-small molecule, provides critical insights into cellular processes and is essential for the development of drug diagnostics and therapeutics. Here we present a novel, label-free, and tether-free technology to analyze picomolar to millimolar affinities of biomolecular interactions by Microscale Thermophoresis (MST). The entropy of the hydration shell surrounding molecules determines thermophoretic movement. MST exploits this principle by measuring interactions using optically generated temperature gradients. MST detects changes in the size, charge and hydration shell of molecules and measures biomolecule interactions under close-to-native conditions: immobilization-free and in bioliquids of choice, including cell lysates and blood serum. Thus, MST measures interactions under close-to-native conditions, and without laborious sample purification. We demonstrate how MST determines the picomolar affinities of antibody::antigen interactions, and protein::protein interactions measured from directly from cell lysates. MST assays are highly adaptable to fit to the diverse requirements of different and complex biomolecules. NanoTemper´s unique technology is ideal for studies requiring flexibility and sensitivity at the experimental scale, making MST suitable for basic research investigations and pharmaceutical applications.

Keywords: biochemistry, biophysics, molecular interactions, quantitative techniques

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Authors: Salina Yahya Saddick

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Ovarian cancer remains the most lethal gynecological malignancy due to the lack of highly sensitive and specific screening tools for detection of early-stage disease. The OSE provides the progenitor cells for 90% of human ovarian cancers. Recent morphologic, immunohistochemical and molecular genetic studies have led to the development of a new paradigm for the pathogenesis and origin of epithelial ovarian cancer (EOC) based on a ualistic model of carcinogenesis that divides EOC into two broad categories designated Types I and II which are characterized by specific mutations, including KRAS, BRAF, ERBB2, CTNNB1, PTEN PIK3CA, ARID1A, and PPPR1A, which target specific cell signaling pathways. Type 1 tumors rarely harbor TP53. type I tumors are relatively genetically stable and typically display a variety of somatic sequence mutations that include KRAS, BRAF, PTEN, PIK3CA CTNNB1 (the gene encoding beta catenin), ARID1A and PPP2R1A but very rarely TP53 . The cancer stem cell (CSC) hypothesis postulates that the tumorigenic potential of CSCs is confined to a very small subset of tumor cells and is defined by their ability to self-renew and differentiate leading to the formation of a tumor mass. Potential protein biomarker miRNA, are promising biomarkers as they are remarkably stable to allow isolation and analysis from tissues and from blood in which they can be found as free circulating nucleic acids and in mononuclear cells. Recently, genomic anaylsis have identified biomarkers and potential therapeutic targets for ovarian cancer namely, FGF18 which plays an active role in controlling migration, invasion, and tumorigenicity of ovarian cancer cells through NF-κB activation, which increased the production of oncogenic cytokines and chemokines. This review summarizes update information on epithelial ovarian cancers and point out to the most recent ongoing research.

Keywords: epithelial ovarian cancers, somatic sequence mutations, cancer stem cell (CSC), potential protein, biomarker, genomic analysis, FGF18 biomarker

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Authors: E. Nakamachi, S. Tanaka, K. Yamamoto, Y. Morita

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In this study, we developed a three-dimensional (3D) direct current electric field (DCEF) stimulation bio-reactor for axonal outgrowth enhancement to generate the neural network of the central nervous system (CNS). By using our newly developed 3D DCEF stimulation bio-reactor, we cultured the rat pheochromocytoma cells (PC12) and investigated the effects on the axonal extension enhancement and network generation. Firstly, we designed and fabricated a 3D bio-reactor, which can load DCEF stimulation on PC12 cells embedded in the collagen gel as extracellular environment. The connection between the electrolyte and the medium using salt bridges for DCEF stimulation was introduced to avoid the cell death by the toxicity of metal ion. The distance between the salt bridges was adopted as the design variable to optimize a structure for uniform DCEF stimulation, where the finite element (FE) analyses results were used. Uniform DCEF strength and electric flux vector direction in the PC12 cells embedded in collagen gel were examined through measurements of the fabricated 3D bio-reactor chamber. Measurement results of DCEF strength in the bio-reactor showed a good agreement with FE results. In addition, the perfusion system was attached to maintain pH 7.2 ~ 7.6 of the medium because pH change was caused by DCEF stimulation loading. Secondly, we disseminated PC12 cells in collagen gel and carried out 3D culture. Finally, we measured the morphology of PC12 cell bodies and neurites by the multiphoton excitation fluorescence microscope (MPM). The effectiveness of DCEF stimulation to enhance the axonal outgrowth and the neural network generation was investigated. We confirmed that both an increase of mean axonal length and axogenesis rate of PC12, which have been exposed 5 mV/mm for 6 hours a day for 4 days in the bioreactor. We found following conclusions in our study. 1) Design and fabrication of DCEF stimulation bio-reactor capable of 3D culture nerve cell were completed. A uniform electric field strength of average value of 17 mV/mm within the 1.2% error range was confirmed by using FE analyses, after the structure determination through the optimization process. In addition, we attached a perfusion system capable of suppressing the pH change of the culture solution due to DCEF stimulation loading. 2) Evaluation of DCEF stimulation effects on PC12 cell activity was executed. The 3D culture of PC 12 was carried out adopting the embedding culture method using collagen gel as a scaffold for four days under the condition of 5.0 mV/mm and 10mV/mm. There was a significant effect on the enhancement of axonal extension, as 11.3% increase in an average length, and the increase of axogenesis rate. On the other hand, no effects on the orientation of axon against the DCEF flux direction was observed. Further, the network generation was enhanced to connect longer distance between the target neighbor cells by DCEF stimulation.

Keywords: PC12, DCEF stimulation, 3D bio-reactor, axonal extension, neural network generation

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Authors: Chang-Hui Shen, Paulina Konarzewska

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Cell homeostasis is regulated by vacuolar activity and it has been shown that lipid composition of the vacuole plays an important role in vacuolar function. The major phosphoinositide species present in the vacuolar membrane include phosphatidylinositol 3,5-biphosphate (PI(3,5)P₂) which is generated from PI(3)P controlled by Fab1p. Deletion of FAB1 gene reduce the synthesis of PI(3,5)P₂ and thus result in enlarged or fragmented vacuoles, with neutral vacuolar pH due to reduced vacuolar H⁺-ATPase activity. These mutants also exhibited poor growth at high extracellular pH and in the presence of CaCl₂. Conversely, VPS34 regulates the synthesis of PI(3)P from phosphatidylinositol (PI), and the lack of Vps34p results in the reduction of vacuolar activity. Although the cellular observations are clear, it is still unknown about the molecular mechanism between the phospholipid biosynthesis pathway and vacuolar activity. Since both VPS34 and FAB1 are important in vacuolar activity, we hypothesize that the molecular mechanism of vacuolar function might be regulated by the transcriptional regulators of phospholipid biosynthesis. In this study, we study the role of the major phospholipid biosynthesis transcription factor, INO2, in the regulation of vacuolar activity. We first performed qRT-PCR to examine the effect of Ino2p on the expression of VPS34 and FAB1. Our results showed that VPS34 was upregulated in the presence of inositol for both WT and ino2Δ cells. However, FAB1 was only upregulated significantly in ino2Δ cells. This indicated that Ino2p might be the negative regulator for FAB1 expression. Next, growth sensitivity experiment showed that WT, vma3Δ, and ino2Δ grew well in growth medium buffered to pH 5.5 containing 10 mM CaCl₂. As cells were switched to growth medium buffered to pH 7 containing CaCl₂ WT, ino2Δ and opi1Δ showed growth reduction, whereas vma3Δ was completely nonviable. As the concentration of CaCl₂ was increased to 60 mM, ino2Δ cells showed moderate growth reduction compared to WT. This result suggests that ino2Δ cells have better vacuolar activity. Microscopic analysis and vacuolar acidification were employed to further elucidate the importance of INO2 in vacuolar homeostasis. Analysis of vacuolar morphology indicated that WT and vma3Δ cells displayed vacuoles that occupied a small area of the cell when grown in media buffered to pH 5.5. Whereas, ino2Δ displayed fragmented vacuoles. On the other hand, all strains grown in media buffered to pH 7, exhibited enlarged vacuoles that occupied most of the cell’s surface. This indicated that the presence of INO2 may play negative effect in vacuolar morphology when cells are grown in media buffered to pH 5.5. Furthermore, vacuolar acidification assay showed that only vma3Δ cells displayed notably less acidic vacuoles as cells were grown in media buffered to pH 5.5 and pH 7. Whereas, ino2Δ cells displayed more acidic pH compared to WT at pH7. Taken together, our results demonstrated the molecular mechanism of the vacuolar activity regulated by the phospholipid biosynthesis transcription factors Ino2p. Ino2p negatively regulates vacuolar activity through the expression of FAB1.

Keywords: vacuole, phospholipid, homeostasis, Ino2p, FAB1

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Authors: Ethan Nikookar, Yalda Boroushaki, Larissa Statsenko, Jorge Ochoa Paniagua

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Given the serious impact that supply chain disruptions can have on a firm's bottom-line performance, both industry and academia are interested in supply chain resilience, a capability of the supply chain that enables it to cope with disruptions. To date, much of the research has focused on the antecedents of supply chain resilience. This line of research has suggested various firm-level capabilities that are associated with greater supply chain resilience. A consensus has emerged among researchers that supply chain flexibility holds the greatest potential to create resilience. Supply chain flexibility achieves resilience by creating readiness to respond to disruptions with little cost and time by means of reconfiguring supply chain resources to mitigate the impacts of the disruption. Decisions related to supply chain disruptions are made by supply chain managers; however, the role played by supply chain managers' reference networks has been overlooked in the supply chain resilience literature. This study aims to understand the impact of supply chain managers on their firms' supply chain resilience. Drawing on social capital theory and social network theory, this paper proposes a conceptual model to explore the role of supply chain managers in developing the resilience of supply chains. Our model posits that higher level of supply chain managers' embeddedness in their reference network is associated with increased resilience of their firms' supply chain. A reference network includes individuals from whom supply chain managers seek advice on supply chain related matters. The relationships between supply chain managers' embeddedness in reference network and supply chain resilience are mediated by supply chain flexibility.

Keywords: supply chain resilience, embeddedness, reference networks, social capitals

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Authors: Ayda Youssef, Tarek Rafaat, Iman zaky

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Purpose: Langerhans’ cell histiocytosis (LCH) is not uncommon pathology that implies aberrant proliferation of a specific dendritic (Langerhans) cell. These atypical but mature cells of monoclonal origin can infiltrate many sites of the body and may occur as localized lesions or as widespread systemic disease. Liver is one of the uncommon sites of affection. The twofold objective of this study is to illustrate the radiological presentation of this disease, and to compare these results with previously reported series. Methods and Materials: Between 2007 and 2012, 150 patients with biopsy-proven LCH were treated in our hospital, a paediatric cancer tertiary care center. A retrospective review of radiographic images and reports was performed. There were 33 patients with liver affection are stratified. All patients underwent imaging studies, mostly US and CT. A chart review was performed to obtain demographic, clinical and radiological data. They were analyzed and compared to other published series. Results: Retrospective assessment of 150 patients with LCH was performed, among them 33 patients were identified who had liver involvement. All these patients developed multisystemic disease; They were 12 females and 21 males with (n= 32), seven of them had marked hepatomegaly. Diffuse hypodense liver parenchyma was encountered in five cases, the periportal location has a certain predilection in cases of focal affection where three cases has a hypodense periportal soft tissue sheets, one of them associated with dilated biliary radicals, only one case has multiple focal lesions unrelated to portal tracts. On follow up of the patients, two cases show abnormal morphology of liver with bossy outline. Conclusion: LCH is a not infrequent disease. A high-index suspicion should be raised in the context of diagnosis of liver affection. A biopsy is recommended in the presence of radiological suspicion. Chemotherapy is the preferred therapeutic modality. Liver histiocytosis are not disease specific features but should be interpreted in conjunction with the clinical history and the results of biopsy. Clinical Relevance/Application: Radiologist should be aware of different patterns of hepatobiliary histiocytosis, Thus early diagnosis and proper management of patient can be conducted.

Keywords: langerhans’ cell histiocytosis, liver, medical and health sciences, radiology

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Authors: Anna Cameron, Chunxia Zhao, Haofei Wang, Yun Liu, Guang Ze Yang

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Since the first publication in 1775, cancer research has built a comprehensive understanding of how cellular components of the tumor niche promote disease development. However, only within the last decade has research begun to establish the impact of non-cellular components of the niche, particularly the extracellular matrix (ECM). The ECM, a three-dimensional scaffold that sustains the tumor microenvironment, plays a crucial role in disease progression. Cancer cells actively deregulate and remodel the ECM to establish a tumor-promoting environment. Recent work has highlighted the need to further our understanding of the complexity of this cancer-ECM relationship. In vitro models use hydrogels to mimic the ECM, as hydrogel matrices offer biological compatibility and stability needed for long term cell culture. However, natural hydrogels are being used in these models verbatim, without tuning their biophysical characteristics to achieve pathophysiological relevance, thus limiting their broad use within cancer research. The biophysical attributes of these gels dictate cancer cell proliferation, invasion, metastasis, and therapeutic response. Evaluating the three most widely used natural hydrogels, Matrigel, collagen, and agarose gel, the permeability, stiffness, and pore-size of each gel were measured and compared to the in vivo environment. The pore size of all three gels fell between 0.5-6 µm, which coincides with the 0.1-5 µm in vivo pore size found in the literature. However, the stiffness for hydrogels able to support cell culture ranged between 0.05 and 0.3 kPa, which falls outside the range of 0.3-20,000 kPa reported in the literature for an in vivo ECM. Permeability was ~100x greater than in vivo measurements, due in large part to the lack of cellular components which impede permeation. Though, these measurements prove important when assessing therapeutic particle delivery, as the ECM permeability decreased with increasing particle size, with 100 nm particles exhibiting a fifth of the permeability of 10 nm particles. This work explores ways of adjusting the biophysical characteristics of hydrogels by changing protein concentration and the trade-off, which occurs due to the interdependence of these factors. The global aim of this work is to produce a more pathophysiologically relevant model for each tumor type.

Keywords: cancer, extracellular matrix, hydrogel, microfluidic

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

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

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

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Authors: Mengfei Peng, Serajus Salaheen, Debabrata Biswas

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Lactobacillus casei found in the human intestine and mouth is commonly applied for dairy production. Recently, it was found that some byproducts produced by Lactobacillus exhibited antimicrobial activities against multiple bacteria. Meanwhile, introduction of prebiotic-like foods (e.g. cocoa) or probiotics or both of them as food supplements in human diets as well as in farm animal feeds is believed to be an effective ways in control/reduce the colonization of foodborne bacterial pathogens infection in the gut environment. We hypothesized that cocoa may stimulate the production antimicrobial components of Lactobacillus casei and may potentially inhibit/reduce the colonization and infection of foodborne bacterial pathogens in the gut. Mixed culture of L. casei (LC) with enterohemorrhagic E. coli EDL933 (EHEC), Salmonella Typhimurium LT2 (ST), or Listeria monocytogenes LM2 (LM) showed that LC could competitively exclude (100%) them within 72 h. Further, investigation of cell-free culture supernatant (CFCS) revealed that the antimicrobial effects of LC came from CFCS. CFCS of LC eliminated (100%) EHEC, ST, and LM within 72 h, and 2 h CFCS treatment increased the hydrophobicity of EHEC (5.10 folds), ST (8.48 folds), and LM (2.03 folds). In addition, LC cells exhibited more inhibitive effects than CFCS on cell adhesive and invasive activities of EHEC (52.14% & 90.45%), ST (66.89% & 93.83%), and LM (61.10% & 83.40%). Two clusters of poly-peptides in CFCS were identified by SDS-PAGE, the molecular weights of which are ≈5 KD and 40-45 KD. LC CFCS with overnight growth in the presence of 3% strengthened all of the antimicrobial activities (growth inhibition, outer membrane disruption, and cell infective ability reduction). Liquid chromatography/Mass spectrometry analysis detected 5 unique components in class of flavonoids in LC CFCS with overnight 3% cocoa supplement. Furthermore, qPCR results showed that CFCSs up-regulated the expression level of genes responsible for flagellin synthesis and motility, but down-regulated genes for specific binding and invasion-associated proteins synthesis. The stimulatory effects of cocoa in producing bioactive components of probiotics may aid prevention of foodborne illness caused by major foodborne enteric bacterial pathogens.

Keywords: foodborne pathogens, probiotics, prebiotics, pathogen exclusion

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Authors: R. Marhenke, M. Martini

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The distraction of attentional resources after learning hinders long-term memory consolidation compared to several minutes of post-encoding inactivity in form of wakeful resting. We tested whether an 8-minute period of wakeful resting, compared to performing an adapted version of the d2 test of attention after learning, supports memory retention. Participants encoded and immediately recalled a word list followed by either an 8 minute period of wakeful resting (eyes closed, relaxed) or by performing an adapted version of the d2 test of attention (scanning and selecting specific characters while ignoring others). At the end of the experimental session (after 12-24 min) and again after 7 days, participants were required to complete a surprise free recall test of both word lists. Our results showed no significant difference in memory retention between the experimental conditions. However, we found that participants who completed the first lines of the d2 test in less than the given time limit of 20 seconds and thus had short unfilled intervals before switching to the next test line, remembered more words over the 12-24 minute and over the 7 days retention interval than participants who did not complete the first lines. This interaction occurred only for the first test lines, with the highest temporal proximity to the encoding task and not for later test lines. Differences in retention scores between groups (completed first line vs. did not complete) seem to be widely independent of the general performance in the d2 test. Implications and limitations of these exploratory findings are discussed.

Keywords: long-term memory, retroactive interference, attention, forgetting

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Authors: Xin Chen, Xinyong Li, Qidong Zhao, Dong Wang

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A new coupling system was constructed by combining photo-electrochemical cell with electro-fenton cell (PEC-EF). The electrode material in this system was derived from MnyFe₁₋yCo Prussian-Blue-Analog (PBA). Mn₀.₄Fe₀.₆Co₀.₆₇-N@C spin-coated on carbon paper behaved as the gas diffusion cathode and Mn₀.₄Fe₀.₆Co₀.₆₇O₂.₂ spin-coated on fluorine-tin oxide glass (FTO) as anode. The two separated cells could degrade Sulfamethoxazole (SMX) simultaneously and some coupling mechanisms by PEC and EF enhancing the degradation efficiency were investigated. The continuous on-site generation of H₂O₂ at cathode through an oxygen reduction reaction (ORR) was realized over rotating ring-disk electrode (RRDE). The electron transfer number (n) of the ORR with Mn₀.₄Fe₀.₆Co₀.₆₇-N@C was 2.5 in the selected potential and pH range. The photo-electrochemical properties of Mn₀.₄Fe₀.₆Co₀.₆₇O₂.₂ were systematically studied, which displayed good response towards visible light. The photoinduced electrons at anode can transfer to cathode for further use. Efficient photo-electro-catalytic performance was observed in degrading SMX. Almost 100% SMX removal was achieved in 120 min. This work not only provided a highly effective technique for antibiotic treatment but also revealed the synergic effect between PEC and EF.

Keywords: electro-fenton, photo-electrochemical, synergic effect, sulfamethoxazole

Procedia PDF Downloads 178

Authors: Hela Ghali, Oumayma Ben Amor, Salwa Khefacha, Mohamed Ben Rejeb, Sirine Frigui, Meriam Tourki Dhidah, Lamine Dhidah, Houyem Said Laatiri

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Background: Blood exposure accidents are the most common problem in hospitals that threaten healthcare professionals with a high risk of infectious complications which weighs heavily on health systems worldwide. Paramedics are the highest risk group due to the nature of their daily activities. We aimed to determine knowledge and attitudes about the management of blood-exposure accidents among nurses and technicians in two pediatric departments. Materials/Methods: This is a cross-sectional descriptive study conducted on March 2017, carried out with the care staff of the pediatric ward of the Farhat Hached Teaching Hospital of Sousse and pediatric surgery of the Fattouma Bourguiba University Hospital in Monastir, using a pre- tested and self-administered questionnaire. Data entry and analysis were performed using Excel software. Results: The response rate was 85.1%. A female predominance (82.5%) was reported among respondents with a sex ratio of 0.21. 80% of the participants were under 35 years old. Seniority of less than 10 years was found in 77.5% of respondents. Only 22.5% knew the definition of a blood- exposure accident. 100% and 95% of participants reported the relative risk, respectively, to hepatitis and AIDS viruses. However, only 15% recognized the severity factors of a blood-exposure accident. Hygiene compliance was the most important dimension for almost the entire population for the prevention. On the other hand, only 12.5% knew the meaning of 'standard precautions' and ¼ considered them necessary for at-risk patients only. 40% reported being exposed at least once, among them, 87.5% used betadine, and 77.5% said that anti-infectious chemoprophylaxis is necessary regardless of the patient's serological status. However, 52.5% did not know the official reporting circuit of management of blood-exposure accident in their institutions. Conclusion: For better management of risks in hospitals and an improvement of the safety of the care, a reinforcement of the sensibilization of the caregivers with regard to the risks of blood exposure accident is necessary, while developing their knowledge to act in security.

Keywords: attitudes, blood-exposure accident, knowledge, pediatric department

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Authors: Huazhen Lin, Ruihua Xu, Zhibin Jiang

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In this real-world case, timetabling the LRT network as a whole is rather challenging for the operator: they are supposed to create a timetable to avoid various route conflicts manually while satisfying a given interval and the number of rolling stocks, but the outcome is not satisfying. Therefore, the operator adopts a computerised timetabling tool, the Train Plan Maker (TPM), to cope with this problem. However, with various constraints in the dual-line network, it is still difficult to find an adequate pairing of turnback time, interval and rolling stocks’ number, which requires extra manual intervention. Aiming at current problems, a one-off model for timetabling is presented in this paper to simplify the procedure of timetabling. Before the timetabling procedure starts, this paper presents how the dual-line system with a ring and several branches is turned into a simpler structure. Then, a non-linear programming model is presented in two stages. In the first stage, the model sets a series of constraints aiming to calculate a proper timing for coordinating two lines by adjusting the turnback time at termini. Then, based on the result of the first stage, the model introduces a series of inequality constraints to avoid various route conflicts. With this model, an analysis is conducted to reveal the relation between the ratio of trains in different directions and the possible minimum interval, observing that the more imbalance the ratio is, the less possible to provide frequent service under such strict constraints.

Keywords: light rail transit (LRT), non-linear programming, railway timetabling, timetable coordination

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Authors: Ali Mirtaghavi, Andy Baldwin, Rajendarn Muthuraj, Jack Luo

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Recent advances in biomaterials have led to utilizing biopolymers to develop 3D scaffolds in tissue regeneration. One of the major challenges of designing biomaterials for 3D scaffolds is to mimic the building blocks similar to the extracellular matrix (ECM) of the native tissues. Biopolymer based aerogels obtained by freeze-drying have shown to provide structural similarities to the ECM owing to their 3D format and a highly porous structure with interconnected pores, similar to the ECM. Gelatin (GEL) is known to be a promising biomaterial with inherent regenerative characteristics owing to its chemical similarities to the ECM in native tissue, biocompatibility abundance, cost-effectiveness and accessible functional groups, which makes it facile for chemical modifications with other biomaterials to form biocomposites. Despite such advantages, gelatin offers poor mechanical properties, sensitive enzymatic degradation and high viscosity at room temperature which limits its application and encourages its use to develop biocomposites. Hydrophilic biomass-based cellulose nanofibrous (CNF) has been explored to use as suspension for biocomposite aerogels for the development of 3D porous structures with excellent mechanical properties, biocompatibility and slow enzymatic degradation. In this work, CNF biocomposite aerogels with various ratios of CNF:GEL) (90:10, 70:30 and 50:50) were prepared by freeze-drying technique, and their properties were investigated in terms of physicochemical, mechanical and biological characteristics. Epichlorohydrin (EPH) was used to investigate the effect of chemical crosslinking on the molecular interaction of CNF: GEL, and its effects on physicochemical, mechanical and biological properties of the biocomposite aerogels. Ultimately, chemical crosslinking helped to improve the mechanical resilience of the resulting aerogels. Amongst all the CNF-GEL composites, the crosslinked CNF: GEL (70:30) biocomposite was found to be favourable for cell attachment and viability. It possessed highly porous structure (porosity of ~93%) with pore sizes ranging from 16-110 µm, adequate mechanical properties (compression modulus of ~47 kPa) and optimal biocompatibility both in-vitro and in-vivo, as well as controlled enzymatic biodegradation, high water penetration, which could be considered a suitable option for wound healing application. In-vivo experiments showed improvement on inflammation and foreign giant body cell reaction for the crosslinked CNF: GEL (70:30) compared to the other samples. This could be due to the superior interaction of CNF with gelatin through chemical crosslinking, resulting in more optimal in-vivo improvement. In-vitro cell culture investigation on human dermal fibroblasts showed satisfactory 3D cell attachment over time. Overall, it has been observed that the developed CNF: GEL aerogel can be considered as a potential scaffold for soft tissue regeneration application.

Keywords: 3D scaffolds, aerogels, Biocomposites , tissue engineering

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Authors: Ozlem Oral, Emre Taskin, Aysel Yuce, Serap Dokmeci, Devrim Gozuacik

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Autophagy is an evolutionary conserved lysosome-dependent catabolic pathway, responsible for the degradation of long-lived proteins, abnormal aggregates and damaged organelles which cannot be degraded by the ubiquitin-proteasome system. Lysosomes degrade the substrates through the activity of lysosomal hydrolases and lysosomal membrane-bound proteins. Mutations in the coding region of these proteins cause malfunctional lysosomes, which contributes to the pathogenesis of lysosomal storage diseases. Gaucher disease is a lysosomal storage disease resulting from the mutation of a lysosomal membrane-associated glycoprotein called glucocerebrosidase and its cofactor saposin C. The disease leads to intracellular accumulation of glucosylceramide and other glycolipids. Because of the essential role of lysosomes in autophagic degradation, Gaucher disease may directly be linked to this pathway. In this study, we investigated the expression of autophagy and/or lysosome-related genes and proteins in fibroblast cells isolated from patients with different mutations. We carried out confocal microscopy analysis and examined autophagic flux by utilizing the differential pH sensitivities of RFP and GFP in mRFP-GFP-LC3 probe. We also evaluated lysosomal pH by active lysosome staining and lysosomal enzyme activity. Beside lysosomes, we also performed proteasomal activity and cell death analysis in patient samples. Our data showed significant attenuation in the expression of key autophagy-related genes and accumulation of their proteins in mutant cells. We found decreased the ability of autophagosomes to fuse with lysosomes, associated with elevated lysosomal pH and reduced lysosomal enzyme activity. Proteasomal degradation and cell death analysis showed reduced proteolytic activity of the proteasome, which consequently leads to increased susceptibility to cell death. Our data indicate that the major degradation pathways are affected by multifunctional lysosomes in mutant patient cells and may underlie in the mechanism of clinical severity of Gaucher patients. (This project is supported by TUBITAK-3501-National Young Researchers Career Development Program, Project No: 112T130).

Keywords: autophagy, Gaucher's disease, glucocerebrosidase, mutant fibroblasts

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Authors: Sang Chul Lee, Gi-Young Park, Dong Rak Kwon

Abstract:

Objective: The aim of this study was to investigate regenerative effects of ultrasound (US)-guided injection with human umbilical cord blood-derived mesenchymal stem cells (UCB-MSCs) and/or polydeoxyribonucleotide (PDRN) injection in a chronic traumatic full-thickness rotator cuff tendon tear (FTRCTT) in a rabbit model. Material and Methods: Rabbits (n = 32) were allocated into 4 groups. After a 5-mm sized FTRCTT just proximal to the insertion site on the subscapularis tendon was created by excision, the wound was immediately covered by silicone tube to prevent natural healing. After 6 weeks, 4 injections (0.2 mL normal saline, G1; 0.2 mL PDRN, G2; 0.2 mL UCB-MSCs, G3; and 0.2 mL UCB-MSCs with 0.2ml PDRN, G4) were injected into FTRCTT under US guidance. We evaluated gross morphologic changes on all rabbits after sacrifice. Masson’s trichrome, anti-type 1 collagen antibody, bromodeoxyuridine, proliferating cell nuclear antigen, vascular endothelial growth factor and platelet endothelial cell adhesion molecule stain were performed to evaluate histological changes. Motion analysis was also performed. Results: The gross morphologic mean tendon tear size in G3 and 4 was significantly smaller than that of G1 and 2 (p < .05). However, there were no significant differences in tendon tear size between G3 and 4. In G4, newly regenerated collagen type 1 fibers, proliferating cells activity, angiogenesis, walking distance, fast walking time, and mean walking speed were greater than in the other three groups on histological examination and motion analysis. Conclusion: Co-injection of UCB-MSCs and PDRN was more effective than UCB-MSCs injection alone in histological and motion analysis in a rabbit model of chronic traumatic FTRCTT. However, there was no significant difference in gross morphologic change of tendon tear between UCB-MSCs with/without PDRN injection. The results of this study regarding the combination of UCB-MSCs and PDRN are worth additional investigations.

Keywords: mesenchymal stem cell, umbilical cord, polydeoxyribonucleotides, shoulder, rotator cuff, ultrasonography, injections

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