Search results for: Epithelial Transport
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 2095

Search results for: Epithelial Transport

1255 Informal Green Infrastructure as Mobility Enabler in Informal Settlements of Quito

Authors: Ignacio W. Loor

Abstract:

In the context of informal settlements in Quito, this paper provides evidence that slopes and deep ravines typical of Andean cities, around which marginalized urban communities sit, constitute a platform for green infrastructure that supports mobility for pedestrians in an incremental fashion. This is informally shaped green infrastructure that provides connectivity to other mobility infrastructures such as roads and public transport, which permits relegated dwellers reach their daily destinations and reclaim their rights to the city. This is relevant in that walking has been increasingly neglected as a viable mean of transport in Latin American cities, in favor of rather motorized means, for which the mobility benefits of green infrastructure have remained invisible to policymakers, contributing to the progressive isolation of informal settlements. This research leverages greatly on an ecological rejuvenation programme led by the municipality of Quito and the Andean Corporation for Development (CAN) intended for rehabilitating the ecological functionalities of ravines. Accordingly, four ravines in different stages of rejuvenation were chosen, in order to through ethnographic methods, capture the practices they support to dwellers of informal settlements across different stages, particularly in terms of issues of mobility. Then, by presenting fragments of interviews, description of observed phenomena, photographs and narratives published in institutional reports and media, the production process of mobility infrastructure over unoccupied slopes and ravines, and the roles that this infrastructure plays in the mobility of dwellers and their quotidian practices are explained. For informal settlements, which normally feature scant urban infrastructure, mobility embodies an unfavourable driver for the possibilities of dwellers to actively participate in the social, economic and political dimensions of the city, for which their rights to the city are widely neglected. Nevertheless, informal green infrastructure for mobility provides some alleviation. This infrastructure is incremental, since its features and usability gradually evolves as users put into it knowledge, labour, devices, and connectivity to other infrastructures in different dimensions which increment its dependability. This is evidenced in the diffusion of knowledge of trails and routes of footpaths among users, the implementation of linking stairs and bridges, the improved access by producing public spaces adjacent to the ravines, the illuminating of surrounding roads, and ultimately, the restoring of ecological functions of ravines. However, the perpetuity of this type of infrastructure is also fragile and vulnerable to the course of urbanisation, densification, and expansion of gated privatised spaces.

Keywords: green infrastructure, informal settlements, urban mobility, walkability

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1254 The Analysis of Movement Pattern during Reach and Grasp in Stroke Patients: A Kinematic Approach

Authors: Hyo Seon Choi, Ju Sun Kim, DY Kim

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Introduction: This study was aimed to evaluate temporo-spatial patterns during the reach and grasp task in hemiplegic stroke patients and to identify movement pattern according to severity of motor impairment. Method: 29 subacute post-stroke patients were enrolled in this study. The temporo-spatial and kinematic data were obtained during reach and grasp task through 3D motion analysis (VICON). The reach and grasp task was composed of four sub-tasks: reach (T1), transport to mouth (T2), transport back to table (T3) and return (T4). The movement time, joint angle and sum of deviation angles from normative data were compared between affected side and unaffected side. They were also compared between two groups (mild to moderate group: 28~66, severe group: 0~27) divided by upper-Fugl-Meyer Assessment (FMA) scale. Result: In affected side, total time and durations of all four tasks were significantly longer than those in unaffected side (p < 0.001). The affected side demonstrated significant larger shoulder abduction, shoulder internal rotation, wrist flexion, wrist pronation, thoracic external rotation and smaller shoulder flexion during reach and grasp task (p < 0.05). The significant differences between mild to moderate group and severe group were observed in total duration, durations of T1, T2, and T3 in reach and grasp task (p < 0.01). The severe group showed significant larger shoulder internal rotation during T2 (p < 0.05) and wrist flexion during T2, T3 (p < 0.05) than mild to moderate group. In range of motion during each task, shoulder abduction-adduction during T2 and T3, shoulder internal-external rotation during T2, elbow flexion-extension during T1 showed significant difference between two groups (p < 0.05). The severe group had significant larger total deviation angles in shoulder internal-external rotation and wrist extension-flexion during reach and grasp task (p < 0.05). Conclusion: This study suggests that post-stroke hemiplegic patients have an unique temporo-spatial and kinematic patterns during reach and grasp task, and the movement pattern may be related to affected upper limb severity. These results may be useful to interpret the motion of upper extremity in stroke patients.

Keywords: Fugl-Meyer Assessment (FMA), motion analysis, reach and grasp, stroke

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1253 Influence of Smoking on Fine And Ultrafine Air Pollution Pm in Their Pulmonary Genetic and Epigenetic Toxicity

Authors: Y. Landkocz, C. Lepers, P.J. Martin, B. Fougère, F. Roy Saint-Georges. A. Verdin, F. Cazier, F. Ledoux, D. Courcot, F. Sichel, P. Gosset, P. Shirali, S. Billet

Abstract:

In 2013, the International Agency for Research on Cancer (IARC) classified air pollution and fine particles as carcinogenic to humans. Causal relationships exist between elevated ambient levels of airborne particles and increase of mortality and morbidity including pulmonary diseases, like lung cancer. However, due to a double complexity of both physicochemical Particulate Matter (PM) properties and tumor mechanistic processes, mechanisms of action remain not fully elucidated. Furthermore, because of several common properties between air pollution PM and tobacco smoke, like the same route of exposure and chemical composition, potential mechanisms of synergy could exist. Therefore, smoking could be an aggravating factor of the particles toxicity. In order to identify some mechanisms of action of particles according to their size, two samples of PM were collected: PM0.03 2.5 and PM0.33 2.5 in the urban-industrial area of Dunkerque. The overall cytotoxicity of the fine particles was determined on human bronchial cells (BEAS-2B). Toxicological study focused then on the metabolic activation of the organic compounds coated onto PM and some genetic and epigenetic changes induced on a co-culture model of BEAS-2B and alveolar macrophages isolated from bronchoalveolar lavages performed in smokers and non-smokers. The results showed (i) the contribution of the ultrafine fraction of atmospheric particles to genotoxic (eg. DNA double-strand breaks) and epigenetic mechanisms (eg. promoter methylation) involved in tumor processes, and (ii) the influence of smoking on the cellular response. Three main conclusions can be discussed. First, our results showed the ability of the particles to induce deleterious effects potentially involved in the stages of initiation and promotion of carcinogenesis. The second conclusion is that smoking affects the nature of the induced genotoxic effects. Finally, the in vitro developed cell model, using bronchial epithelial cells and alveolar macrophages can take into account quite realistically, some of the existing cell interactions existing in the lung.

Keywords: air pollution, fine and ultrafine particles, genotoxic and epigenetic alterations, smoking

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1252 Development of a Mathematical Model to Characterize the Oil Production in the Federal Republic of Nigeria Environment

Authors: Paul C. Njoku, Archana Swati Njoku

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The study deals with the development of a mathematical model to characterize the oil production in Nigeria. This is calculated by initiating the dynamics of oil production in million barrels revenue plan cost of oil production in million nairas and unit cost of production from 1974-1982 in the contest of the federal Republic of Nigeria. This country export oil to other countries as well as importing specialized crude. The transport network from origin/destination tij to pairs is taking into account simulation runs, optimization have been considered in this study.

Keywords: mathematical oil model development dynamics, Nigeria, characterization barrels, dynamics of oil production

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1251 Influence of Acceptor Dopant on the Physicochemical and Transport Properties of Textured BaCe0.5Zr0.3ln0.2O3−Δ Materials (Ln = Yb, Y, Cd, Sm, Nd)

Authors: J. Lyagaeva, D. Medvedev, A. Brouzgou, A. Demin, P. Tsiakaras

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The investigation of highly conductive and chemically stable electrolytes for solid oxide fuel cells (SOFC) is a necessity. The aim of the present work is to study the influence of acceptor dopant on the functional properties of textured BaCe0.5Zr0.3Ln0.2O3−δ (Ln = Yb, Y, Gd, Sm, Nd) ceramics. The X-Ray diffraction analysis, scanning electron microscopy, dilatometry and 4-probe dc method of conductivity measurements were used. It was found that the mean grain size of ceramics increases (from 1.4 to 3.2 μm), thermal expansion coefficient grows (from 7.6•10–6 to 10.7•10–6 К–1), but ionic conductivity decreases (from 14 to 3 mS cm–1 at 900°С), when ionic radii of impurity acceptor increases from 0.868 Å (Yb3+) to 0.983 Å (Nd3+).

Keywords: acceptor dopant, crystal structure, proton-conducting, SOFC

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1250 Hypersonic Propulsion Requirements for Sustained Hypersonic Flight for Air Transportation

Authors: James Rate, Apostolos Pesiridis

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In this paper, the propulsion requirements required to achieve sustained hypersonic flight for commercial air transportation are evaluated. In addition, a design methodology is developed and used to determine the propulsive capabilities of both ramjet and scramjet engines. Twelve configurations are proposed for hypersonic flight using varying combinations of turbojet, turbofan, ramjet and scramjet engines. The optimal configuration was determined based on how well each of the configurations met the projected requirements for hypersonic commercial transport. The configurations were separated into four sub-configurations each comprising of three unique derivations. The first sub-configuration comprised four afterburning turbojets and either one or two ramjets idealised for Mach 5 cruise. The number of ramjets required was dependent on the thrust required to accelerate the vehicle from a speed where the turbojets cut out to Mach 5 cruise. The second comprised four afterburning turbojets and either one or two scramjets, similar to the first configuration. The third used four turbojets, one scramjet and one ramjet to aid acceleration from Mach 3 to Mach 5. The fourth configuration was the same as the third, but instead of turbojets, it implemented turbofan engines for the preliminary acceleration of the vehicle. From calculations which determined the fuel consumption at incremental Mach numbers this paper found that the ideal solution would require four turbojet engines and two Scramjet engines. The ideal mission profile was determined as being an 8000km sortie based on an averaging of popular long haul flights with strong business ties, which included Los Angeles to Tokyo, London to New York and Dubai to Beijing. This paper deemed that these routes would benefit from hypersonic transport links based on the previously mentioned factors. This paper has found that this configuration would be sufficient for the 8000km flight to be completed in approximately two and a half hours and would consume less fuel than Concord in doing so. However, this propulsion configuration still result in a greater fuel cost than a conventional passenger. In this regard, this investigation contributes towards the specification of the engine requirements throughout a mission profile for a hypersonic passenger vehicle. A number of assumptions have had to be made for this theoretical approach but the authors believe that this investigation lays the groundwork for appropriate framing of the propulsion requirements for sustained hypersonic flight for commercial air transportation. Despite this, it does serve as a crucial step in the development of the propulsion systems required for hypersonic commercial air transportation. This paper provides a methodology and a focus for the development of the propulsion systems that would be required for sustained hypersonic flight for commercial air transportation.

Keywords: hypersonic, ramjet, propulsion, Scramjet, Turbojet, turbofan

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1249 Assessment of Selected Marine Organisms from Malaysian Coastal Areas for Inhibitory Activity against the Chikungunya Virus

Authors: Yik Sin Chan, Nam Weng Sit, Fook Yee Chye, van Ofwegen Leen, de Voogd Nicole, Kong Soo Khoo

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Chikungunya fever is an arboviral disease transmitted by the Aedes mosquitoes. It has resulted in epidemics of the disease in tropical countries in the Indian Ocean and South East Asian regions. The recent spread of this disease to the temperate countries such as France and Italy, coupled with the absence of vaccines and effective antiviral drugs make chikungunya fever a worldwide health threat. This study aims to investigate the anti-chikungunya virus activity of selected marine organism samples collected from Malaysian coastal areas, including seaweeds (Caulerpa racemosa, Caulerpa sertularioides and Kappaphycus alvarezii), a soft coral (Lobophytum microlobulatum) and a sponge (Spheciospongia vagabunda). Following lyophilization (oven drying at 40C for K. alvarezii) and grinding to powder form, each sample was subjected to sequential solvent extraction using hexane, chloroform, ethyl acetate, ethanol, methanol and distilled water in order to extract bioactive compounds. The antiviral activity was evaluated using monkey kidney epithelial (Vero) cells infected with the virus (multiplicity of infection=1). The cell viability was determined by Neutral Red uptake assay. 70% of the 30 extracts showed weak inhibitory activity with cell viability ≤30%. Seven of the extracts exhibited moderate inhibitory activity (cell viability: 31%-69%). These were the chloroform, ethyl acetate, ethanol and methanol extracts of C. racemosa; chloroform and ethyl acetate extracts of L. microlobulatum; and the chloroform extract of C. sertularioides. Only the hexane and ethanol extracts of L. microlobulatum showed strong inhibitory activity against the virus, resulting in cell viabilities (mean±SD; n=3) of 73.3±2.6% and 79.2±0.9%, respectively. The corresponding mean 50% effective concentrations (EC50) for the extracts were 14.2±0.2 and 115.3±1.2 µg/mL, respectively. The ethanol extract of the soft coral L. microlobulatum appears to hold the most promise for further characterization of active principles as it possessed greater selectivity index (SI>5.6) compared to the hexane extract (SI=2.1).

Keywords: antiviral, seaweed, sponge, soft coral, vero cell

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1248 Method for Controlling the Groundwater Polluted by the Surface Waters through Injection Wells

Authors: Victorita Radulescu

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Introduction: The optimum exploitation of agricultural land in the presence of an aquifer polluted by the surface sources requires close monitoring of groundwater level in both periods of intense irrigation and in absence of the irrigations, in times of drought. Currently in Romania, in the south part of the country, the Baragan area, many agricultural lands are confronted with the risk of groundwater pollution in the absence of systematic irrigation, correlated with the climate changes. Basic Methods: The non-steady flow of the groundwater from an aquifer can be described by the Bousinesq’s partial differential equation. The finite element method was used, applied to the porous media needed for the water mass balance equation. By the proper structure of the initial and boundary conditions may be modeled the flow in drainage or injection systems of wells, according to the period of irrigation or prolonged drought. The boundary conditions consist of the groundwater levels required at margins of the analyzed area, in conformity to the reality of the pollutant emissaries, following the method of the double steps. Major Findings/Results: The drainage condition is equivalent to operating regimes on the two or three rows of wells, negative, as to assure the pollutant transport, modeled with the variable flow in groups of two adjacent nodes. In order to obtain the level of the water table, in accordance with the real constraints, are needed, for example, to be restricted its top level below of an imposed value, required in each node. The objective function consists of a sum of the absolute values of differences of the infiltration flow rates, increased by a large penalty factor when there are positive values of pollutant. In these conditions, a balanced structure of the pollutant concentration is maintained in the groundwater. The spatial coordinates represent the modified parameters during the process of optimization and the drainage flows through wells. Conclusions: The presented calculation scheme was applied to an area having a cross-section of 50 km between two emissaries with various levels of altitude and different values of pollution. The input data were correlated with the measurements made in-situ, such as the level of the bedrock, the grain size of the field, the slope, etc. This method of calculation can also be extended to determine the variation of the groundwater in the aquifer following the flood wave propagation in envoys.

Keywords: environmental protection, infiltrations, numerical modeling, pollutant transport through soils

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1247 Analysis of the Homogeneous Turbulence Structure in Uniformly Sheared Bubbly Flow Using First and Second Order Turbulence Closures

Authors: Hela Ayeb Mrabtini, Ghazi Bellakhal, Jamel Chahed

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The presence of the dispersed phase in gas-liquid bubbly flow considerably alters the liquid turbulence. The bubbles induce turbulent fluctuations that enhance the global liquid turbulence level and alter the mechanisms of turbulence. RANS modeling of uniformly sheared flows on an isolated sphere centered in a control volume is performed using first and second order turbulence closures. The sphere is placed in the production-dissipation equilibrium zone where the liquid velocity is set equal to the relative velocity of the bubbles. The void fraction is determined by the ratio between the sphere volume and the control volume. The analysis of the turbulence statistics on the control volume provides numerical results that are interpreted with regard to the effect of the bubbles wakes on the turbulence structure in uniformly sheared bubbly flow. We assumed for this purpose that at low void fraction where there is no hydrodynamic interaction between the bubbles, the single-phase flow simulation on an isolated sphere is representative on statistical average of a sphere network. The numerical simulations were firstly validated against the experimental data of bubbly homogeneous turbulence with constant shear and then extended to produce numerical results for a wide range of shear rates from 0 to 10 s^-1. These results are compared with our turbulence closure proposed for gas-liquid bubbly flows. In this closure, the turbulent stress tensor in the liquid is split into a turbulent dissipative part produced by the gradient of the mean velocity which also contains the turbulence generated in the bubble wakes and a pseudo-turbulent non-dissipative part induced by the bubbles displacements. Each part is determined by a specific transport equation. The simulations of uniformly sheared flows on an isolated sphere reproduce the mechanisms related to the turbulent part, and the numerical results are in perfect accordance with the modeling of the transport equation of the turbulent part. The reduction of second order turbulence closure provides a description of the modification of turbulence structure by the bubbles presence using a dimensionless number expressed in terms of two-time scales characterizing the turbulence induced by the shear and that induced by bubbles displacements. The numerical simulations carried out in the framework of a comprehensive analysis reproduce particularly the attenuation of the turbulent friction showed in the experimental results of bubbly homogeneous turbulence subjected to a constant shear.

Keywords: gas-liquid bubbly flows, homogeneous turbulence, turbulence closure, uniform shear

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1246 Qualitative Modeling of Transforming Growth Factor Beta-Associated Biological Regulatory Network: Insight into Renal Fibrosis

Authors: Ayesha Waqar Khan, Mariam Altaf, Jamil Ahmad, Shaheen Shahzad

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Kidney fibrosis is an anticipated outcome of possibly all types of progressive chronic kidney disease (CKD). Epithelial-mesenchymal transition (EMT) signaling pathway is responsible for production of matrix-producing fibroblasts and myofibroblasts in diseased kidney. In this study, a discrete model of TGF-beta (transforming growth factor) and CTGF (connective tissue growth factor) was constructed using Rene Thomas formalism to investigate renal fibrosis turn over. The kinetic logic proposed by Rene Thomas is a renowned approach for modeling of Biological Regulatory Networks (BRNs). This modeling approach uses a set of constraints which represents the dynamics of the BRN thus analyzing the pathway and predicting critical trajectories that lead to a normal or diseased state. The molecular connection between TGF-beta, Smad 2/3 (transcription factor) phosphorylation and CTGF is modeled using GenoTech. The order of BRN is CTGF, TGF-B, and SMAD3 respectively. The predicted cycle depicts activation of TGF-B (TGF-β) via cleavage of its own pro-domain (0,1,0) and presentation to TGFR-II receptor phosphorylating SMAD3 (Smad2/3) in the state (0,1,1). Later TGF-B is turned off (0,0,1) thereby activating SMAD3 that further stimulates the expression of CTGF in the state (1,0,1) and itself turns off in (1,0,0). Elevated CTGF expression reactivates TGF-B (1,1,0) and the cycle continues. The predicted model has generated one cycle and two steady states. Cyclic behavior in this study represents the diseased state in which all three proteins contribute to renal fibrosis. The proposed model is in accordance with the experimental findings of the existing diseased state. Extended cycle results in enhanced CTGF expression through Smad2/3 and Smad4 translocation in the nucleus. The results suggest that the system converges towards organ fibrogenesis if CTGF remains constructively active along with Smad2/3 and Smad 4 that plays an important role in kidney fibrosis. Therefore, modeling regulatory pathways of kidney fibrosis will escort to the progress of therapeutic tools and real-world useful applications such as predictive and preventive medicine.

Keywords: CTGF, renal fibrosis signaling pathway, system biology, qualitative modeling

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1245 Supergranulation and Its Turbulent Convection

Authors: U. Paniveni

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A few parameters of supergranular cells are studied using intensity patterns from the Kodaikanal Solar Observatory and Dopplergrams from SOHO. The turbulent aspect of the solar supergranulation is established by examining the interrelationships amongst the parameters characterizing a supergranular cell, namely size, lifetime, area, perimeter, fractal dimension, and horizontal flow velocity. The complexity of supergranular cells depicted by their fractal dimension is indicative of their non-laminar characteristics. The findings corroborate Kolmogorov’s theory of turbulence. Some parameters of supergranular cells also show a latitudinal dependence. Supergranulation is a synonym of convective phenomenon and hence can shed light on the physical conditions in the convection zone of the Sun. It plays a major role in the transport and dispersal of magnetic fields that may have a relation to the phases of the solar cycle.

Keywords: sun, granulation, convection, turbulence

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1244 A Study on the Non-Destructive Test Characterization of Carbon Fiber Reinforced Plastics Using Thermo-Graphic Camera

Authors: Hee Jae Shin, In Pyo Cha, Min Sang Lee, Hyun Kyung Yoon, Tae Ho Kim, Yoon Sun Lee, Lee Ku Kwac, Hong Gun Kim

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Non-destructive testing and evaluation techniques for assessing the integrity of composite structures are essential to both reduce manufacturing costs and out of service time of transport means due to maintenance. In this study, Analyze into non-destructive test characterization of carbon fiber reinforced plastics(CFRP) internal and external defects using thermo-graphic camera and transient thermography method. non-destructive testing were characterized by defect size(∅8,∅10,∅12,∅14) and depth(1.2mm,2.4mm).

Keywords: Non-Destructive Test (NDT), thermal characteristic, thermographic camera, Carbon Fiber Reinforced Plastics(CFRP).

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1243 Ensuring Sustainable Urban Mobility in Indian Cities: Need for Creating People Friendly Roadside Public Spaces

Authors: Pushplata Garg

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Mobility, is an integral part of living and sustainability of urban mobility, is essential not only for, but also for addressing global warming and climate change. However, very little is understood about the obstacles/hurdles and likely challenges in the success of plans for sustainable urban mobility in Indian cities from the public perspective. Whereas some of the problems and issues are common to all cities, others vary considerably with financial status, function, the size of cities and culture of a place. Problems and issues similar in all cities relate to availability, efficiency and safety of public transport, last mile connectivity, universal accessibility, and essential planning and design requirements of pedestrians and cyclists are same. However, certain aspects like the type of means of public transportation, priority for cycling and walking, type of roadside activities, are influenced by the size of the town, average educational and income level of public, financial status of the local authorities, and culture of a place. The extent of public awareness, civic sense, maintenance of public spaces and law enforcement vary significantly from large metropolitan cities to small and medium towns in countries like India. Besides, design requirements for shading, location of public open spaces and sitting areas, street furniture, landscaping also vary depending on the climate of the place. Last mile connectivity plays a major role in success/ effectiveness of public transport system in a city. In addition to the provision of pedestrian footpaths connecting important destinations, sitting spaces and necessary amenities/facilities along footpaths; pedestrian movement to public transit stations is encouraged by the presence of quality roadside public spaces. It is not only the visual attractiveness of streetscape or landscape or the public open spaces along pedestrian movement channels but the activities along that make a street vibrant and attractive. These along with adequate spaces to rest and relax encourage people to walk as is observed in cities with successful public transportation systems. The paper discusses problems and issues of pedestrians for last mile connectivity in the context of Delhi, Chandigarh, Gurgaon, and Roorkee- four Indian cities representing varying urban contexts, that is, of metropolitan, large and small cities.

Keywords: pedestrianisation, roadside public spaces, last mile connectivity, sustainable urban mobility

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1242 Characterization of a Mesenchymal Stem Cells Pool in Killian Nasal Polyp

Authors: Emanuela Chiarella, Clelia Nisticò, Nicola Lombardo, Giovanna Lucia Piazzetta, Nadia Lobello, Maria Mesuraca

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Killian’s Antrochoanal Polyp is a benign lesion of the maxillary sinus characterized by unilateral nasal obstruction, pus discharge, and headache. It affects, more commonly children and young adults. Although its etiology still remains unclear, chronic inflammation, autoreactivity, allergies, and viral infections are strongly associated with its formation and development, resulting in nasal tissue remodeling. We aimed to investigate the stem cells components which reside in this pathological tissue. In particular, we adopted a protocol for the isolation and culturing of mesenchymal stem cells from surgical biopsies of three Killian nasal polyp patients (KNP-MSCs) as well as from their healthy nasal tissue (HNT-MSCs) that were used as controls. The immunophenotype profile of HNT-MSCs and KNP-MSCs was more similar, with a marked positivity for CD73, CD90, and CD105 expression, while being negative for CD34 and CD14 haematopoietic genes. Cell proliferation assay showed that KNP-MSCs had a replicative disadvantage compared to HNT-MSCs, as evidenced by the significantly lower number of cells in the S-phase of the cell cycle. KNP-MSCs also took longer to close a wound than HNT-MSCs, indicating a partial epithelial phenotype in which low levels of ICAM-1 mRNA and a significant increase in E-CAD transcript were detectable. Subsequently, the differentiation potential of both MSCs populations was analyzed by inducing osteoblastic or adipocyte differentiation for up to 20 days. KNP-MSCs showed the ability to differentiate into osteoblasts, although ALP activity as well as the number and size of calcium deposits were lower than osteogenic induced-HNT-MSCs. Also, mRNA levels of osteoblastic marker genes (OCN, OPN, OSX, RUNX2) resulted lower compared to control cell population. Instead, the analysis of the adipogenic differentiation potential showed a similar behavior between KNP-MSCs and HNT-MSCs considering that the amount of lipid droplets, the expression of adipocyte-specific genes (FABP4, AdipoQ, PPARγ2, LPL) and the content of triacylglycerols were almost overlapping. Taken together, these results first demonstrated that Killian's nasal polyp is a source of mesenchymal stem cells with self-renewal and multi-differentiative capabilities.

Keywords: Mesenchymal stem cells, adipogenic differentiation, osteogenic differentiation, EMT

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1241 A Study on the Quantitative Evaluation Method of Asphalt Pavement Condition through the Visual Investigation

Authors: Sungho Kim, Jaechoul Shin, Yujin Baek

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In recent years, due to the environmental impacts and time factor, etc., various type of pavement deterioration is increasing rapidly such as crack, pothole, rutting and roughness degradation. The Ministry of Land, Infrastructure and Transport maintains regular pavement condition of the highway and the national highway using the pavement condition survey equipment and structural survey equipment in Korea. Local governments that maintain local roads, farm roads, etc. are difficult to maintain the pavement condition using the pavement condition survey equipment depending on economic conditions, skills shortages and local conditions such as narrow roads. This study presents a quantitative evaluation method of the pavement condition through the visual inspection to overcome these problems of roads managed by local governments. It is difficult to evaluate rutting and roughness with the naked eye. However, the condition of cracks can be evaluated with the naked eye. Linear cracks (m), area cracks (m²) and potholes (number, m²) were investigated with the naked eye every 100 meters for survey the cracks. In this paper, crack ratio was calculated using the results of the condition of cracks and pavement condition was evaluated by calculated crack ratio. The pavement condition survey equipment also investigated the pavement condition in the same section in order to evaluate the reliability of pavement condition evaluation by the calculated crack ratio. The pavement condition was evaluated through the SPI (Seoul Pavement Index) and calculated crack ratio using results of field survey. The results of a comparison between 'the SPI considering only crack ratio' and 'the SPI considering rutting and roughness either' using the equipment survey data showed a margin of error below 5% when the SPI is less than 5. The SPI 5 is considered the base point to determine whether to maintain the pavement condition. It showed that the pavement condition can be evaluated using only the crack ratio. According to the analysis results of the crack ratio between the visual inspection and the equipment survey, it has an average error of 1.86%(minimum 0.03%, maximum 9.58%). Economically, the visual inspection costs only 10% of the equipment survey and will also help the economy by creating new jobs. This paper advises that local governments maintain the pavement condition through the visual investigations. However, more research is needed to improve reliability. Acknowledgment: The author would like to thank the MOLIT (Ministry of Land, Infrastructure, and Transport). This work was carried out through the project funded by the MOLIT. The project name is 'development of 20mm grade for road surface detecting roadway condition and rapid detection automation system for removal of pothole'.

Keywords: asphalt pavement maintenance, crack ratio, evaluation of asphalt pavement condition, SPI (Seoul Pavement Index), visual investigation

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1240 Numerical Solution of Space Fractional Order Solute Transport System

Authors: Shubham Jaiswal

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In the present article, a drive is taken to compute the solution of spatial fractional order advection-dispersion equation having source/sink term with given initial and boundary conditions. The equation is converted to a system of ordinary differential equations using second-kind shifted Chebyshev polynomials, which have finally been solved using finite difference method. The striking feature of the article is the fast transportation of solute concentration as and when the system approaches fractional order from standard order for specified values of the parameters of the system.

Keywords: spatial fractional order advection-dispersion equation, second-kind shifted Chebyshev polynomial, collocation method, conservative system, non-conservative system

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1239 Modeling of Physico-Chemical Characteristics of Concrete for Filling Trenches in Radioactive Waste Management

Authors: Ilija Plecas, Dalibor Arbutina

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The leaching rate of 60Co from spent mix bead (anion and cation) exchange resins in a cement-bentonite matrix has been studied. Transport phenomena involved in the leaching of a radioactive material from a cement-bentonite matrix are investigated using three methods based on theoretical equations. These are: the diffusion equation for a plane source, an equation for diffusion coupled to a first order equation and an empirical method employing a polynomial equation. The results presented in this paper are from a 25-year mortar and concrete testing project that will influence the design choices for radioactive waste packaging for a future Serbian radioactive waste disposal center.

Keywords: cement, concrete, immobilization, leaching, permeability, radioactivity, waste

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1238 Suspected Odyssean Malaria Outbreak in Gauteng Province, September 2014

Authors: Patience Manjengwa-Hungwe, Carmen White

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Background: Odyssean malaria refers to malaria acquired by infected mosquito bites from malaria endemic to non-endemic regions by mechanical modes of transport, such as airplanes, water vessels, trains and vehicles. Odyssean Malaria is rare and is characterised by absence of travel history to malaria endemic areas. As not anticipated in non-endemic areas, late diagnosis and treatment lead to a high case fatality rate. On 26 September 2014, the Outbreak Response Unit at the National Institute of Communicable Diseases was notified of a suspected death from Odyssean Malaria in Johannesburg, Gauteng Province, a non-endemic area. The main objective of this investigation was to identify the etiological agent's mode and source of transmission. Methods: Epidemiological surveys were conducted with the deceased’s family and clinical details were obtained from doctors who treated the victim in Southrand, Johannesburg. Blood samples were collected prior to death and sent to the National Health Laboratory Services, Johannesburg laboratory for a full blood count, urea electrolytes, creatinine, and C-reactive protein. Environmental assessments and entomological investigations, including collection of mosquito and larvae, were conducted at the deceased’s home and surrounding areas and sent to the laboratory for analysis. Results: Epidemiological surveys revealed no travel history, no mechanical transmission through blood transfusion and no previous possible exposure of the victim to malaria mosquitoes. Laboratory findings indicated that the platelet count was low. A further smear revealed that the malaria parasite was present and malaria antigen for P. falciparum was positive. Entomological findings revealed that none of the six adult or larval mosquitoes collected on site were malaria vectors. Dumping sites found at the back of the house were identified as possible sites where mosquitoes from endemic places could possibly breed. Conclusion: Given that there was no travel history or the possibility of mechanical transmission (blood transfusion or needle), the research team concluded that it is highly probable that the infection was acquired through an infective Anopheles mosquito inadvertently translocated from a Malaria endemic area by mechanical modes of transport. We recommend that clinicians in non-endemic malaria areas be aware of this type of malaria and test for malaria in patients showing malaria-like symptoms.

Keywords: Odyssean Malaria, vector Bourne, malaria, epidemiological surveys

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1237 In vitro Establishment and Characterization of Oral Squamous Cell Carcinoma Derived Cancer Stem-Like Cells

Authors: Varsha Salian, Shama Rao, N. Narendra, B. Mohana Kumar

Abstract:

Evolving evidence proposes the existence of a highly tumorigenic subpopulation of undifferentiated, self-renewing cancer stem cells, responsible for exhibiting resistance to conventional anti-cancer therapy, recurrence, metastasis and heterogeneous tumor formation. Importantly, the mechanisms exploited by cancer stem cells to resist chemotherapy are very less understood. Oral squamous cell carcinoma (OSCC) is one of the most regularly diagnosed cancer types in India and is associated commonly with alcohol and tobacco use. Therefore, the isolation and in vitro characterization of cancer stem-like cells from patients with OSCC is a critical step to advance the understanding of the chemoresistance processes and for designing therapeutic strategies. With this, the present study aimed to establish and characterize cancer stem-like cells in vitro from OSCC. The primary cultures of cancer stem-like cell lines were established from the tissue biopsies of patients with clinical evidence of an ulceroproliferative lesion and histopathological confirmation of OSCC. The viability of cells assessed by trypan blue exclusion assay showed more than 95% at passage 1 (P1), P2 and P3. Replication rate was performed by plating cells in 12-well plate and counting them at various time points of culture. Cells had a more marked proliferative activity and the average doubling time was less than 20 hrs. After being cultured for 10 to 14 days, cancer stem-like cells gradually aggregated and formed sphere-like bodies. More spheroid bodies were observed when cultured in DMEM/F-12 under low serum conditions. Interestingly, cells with higher proliferative activity had a tendency to form more sphere-like bodies. Expression of specific markers, including membrane proteins or cell enzymes, such as CD24, CD29, CD44, CD133, and aldehyde dehydrogenase 1 (ALDH1) is being explored for further characterization of cancer stem-like cells. To summarize the findings, the establishment of OSCC derived cancer stem-like cells may provide scope for better understanding the cause for recurrence and metastasis in oral epithelial malignancies. Particularly, identification and characterization studies on cancer stem-like cells in Indian population seem to be lacking thus provoking the need for such studies in a population where alcohol consumption and tobacco chewing are major risk habits.

Keywords: cancer stem-like cells, characterization, in vitro, oral squamous cell carcinoma

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1236 Multi-Criteria Nautical Ports Capacity and Services Planning

Authors: N. Perko, N. Kavran, M. Bukljas, I. Berbic

Abstract:

This paper is a result of implemented research on proposed introduced methodology for nautical ports capacity planning by introducing a multi-criteria approach of defined criteria and impacts at the Adriatic Sea. The purpose was analysing the determinants -characteristics of infrastructure and services of nautical ports capacity allocated, especially nowadays due to COVID-19 pandemic, as crucial for the successful operation of nautical ports. Giving the importance of the defined priorities for short-term and long-term planning is essential not only in terms of the development of nautical tourism but also in terms of developing the maritime system, but unfortunately, this is not always carried out. Evaluation of the use of resources should follow from a detailed analysis of all aspects of resources bearing in mind that nautical tourism used resources in a sustainable manner and generate effects in the tourism and maritime sectors. Consequently, the identified multiplier effect of nautical tourism, which should be defined and quantified in detail, should be one of the major competitive products on the Croatian Adriatic and the Mediterranean. Research of nautical tourism is necessary to quantify the effects and required planning system development. In the future, the greatest threat to the long-term sustainable development of nautical tourism can be its further uncontrolled or unlimited and undirected development, especially under pressure markedly higher demand than supply for new moorings in the Mediterranean. Results of this implemented research are applicable to nautical ports management and decision-makers of maritime transport system development. This paper will present implemented research and obtained result-developed methodology for nautical port capacity planning -port capacity planning multi-criteria decision-making. A proposed methodological approach of multi-criteria capacity planning includes four criteria (spatial - transport, cost - infrastructure, ecological and organizational criteria, and additional services). The importance of the criteria and sub-criteria is evaluated and carried out as the basis for sensitivity analysis of the importance of the criteria and sub-criteria. Based on the analysis of the identified and quantified importance of certain criteria and sub-criteria, as well as sensitivity analysis and analysis of changes of the quantified importance, scientific and applicable results will be presented. These obtained results have practical applicability by management of nautical ports in the planning of increasing capacity and further development and for the adaptation of existing nautical ports. Obtained research is applicable and replicable in other seas, and results are especially important and useful in this COVID-19 pandemic challenging maritime development framework.

Keywords: Adriatic Sea, capacity, infrastructures, maritime system, methodology, nautical ports, nautical tourism, service

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1235 Immunocytochemical Stability of Antigens in Cytological Samples Stored in In-house Liquid-Based Medium

Authors: Anamarija Kuhar, Veronika Kloboves Prevodnik, Nataša Nolde, Ulrika Klopčič

Abstract:

The decision for immunocytochemistry (ICC) is usually made in the basis of the findings in Giemsa- and/or Papanicolaou- smears. More demanding diagnostic cases require preparation of additional cytological preparations. Therefore, it is convenient to suspend cytological samples in a liquid based medium (LBM) that preserve antigen and morphological properties. However, the duration of these properties being preserved in the medium is usually unknown. Eventually, cell morphology becomes impaired and altered, as well as antigen properties may be lost or become diffused. In this study, the influence of cytological sample storage length in in-house liquid based medium on antigen properties and cell morphology is evaluated. The question is how long the cytological samples in this medium can be stored so that the results of immunocytochemical reactions are still reliable and can be safely used in routine cytopathological diagnostics. The stability of 6 ICC markers that are most frequently used in everyday routine work were tested; Cytokeratin AE1/AE3, Calretinin, Epithelial specific antigen Ep-CAM (MOC-31), CD 45, Oestrogen receptor (ER), and Melanoma triple cocktail were tested on methanol fixed cytospins prepared from fresh fine needle aspiration biopsies, effusion samples, and disintegrated lymph nodes suspended in in-house cell medium. Cytospins were prepared on the day of the sampling as well as on the second, fourth, fifth, and eight day after sample collection. Next, they were fixed in methanol and immunocytochemically stained. Finally, the percentage of positive stained cells, reaction intensity, counterstaining, and cell morphology were assessed using two assessment methods: the internal assessment and the UK NEQAS ICC scheme assessment. Results show that the antigen properties for Cytokeratin AE1/AE3, MOC-31, CD 45, ER, and Melanoma triple cocktail were preserved even after 8 days of storage in in-house LBM, while the antigen properties for Calretinin remained unchanged only for 4 days. The key parameters for assessing detection of antigen are the proportion of cells with a positive reaction and intensity of staining. Well preserved cell morphology is highly important for reliable interpretation of ICC reaction. Therefore, it would be valuable to perform a similar analysis for other ICC markers to determine the duration in which the antigen and morphological properties are preserved in LBM.

Keywords: cytology samples, cytospins, immunocytochemistry, liquid-based cytology

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1234 Cytokine Profiling in Cultured Endometrial Cells after Hormonal Treatment

Authors: Mark Gavriel, Ariel J. Jaffa, Dan Grisaru, David Elad

Abstract:

The human endometrium-myometrium interface (EMI) is the uterine inner barrier without a separatig layer. It is composed of endometrial epithelial cells (EEC) and endometrial stromal cells (ESC) in the endometrium and myometrial smooth muscle cells (MSMC) in the myometrium. The EMI undergoes structural remodeling during the menstruation cycle which are essential for human reproduction. Recently, we co-cultured a layer-by-layer in vitro model of EEC, ESC and MSMC on a synthetic membrane for mechanobiology experiments. We also treated the model with progesterone and β-estradiol in order to mimic the in vivo receptive uterus In the present study we analyzed the cytokines profile in a single layer of EEC the hormonal treated in vitro model of the EMI. The methodologies of this research include simple tissue-engineering . First, we cultured commercial EEC (RL95-2, ATCC® CRL-1671™) in 24-wellplate. Then, we applied an hormonal stimuli protocol with 17-β-estradiol and progesterone in time dependent concentration according to the human physiology that mimics the menstrual cycle. We collected cell supernatant samples of control, pre-ovulation, ovulation and post-ovulaton periods for analysis of the secreted proteins and cytokines. The cytokine profiling was performed using the Proteome Profiler Human XL Cytokine Array Kit (R&D Systems, Inc., USA) that can detect105 human soluble cytokines. The relative quantification of all the cytokines will be analyzed using xMAP – LUMINEX. We conducted a fishing expedition with the 4 membranes Proteome Profiler. We processed the images, quantified the spots intensity and normalized these values by the negative control and reference spots at the membrane. Analyses of the relative quantities that reflected change higher than 5% of the control points of the kit revealed the The results clearly showed that there are significant changes in the cytokine level for inflammation and angiogenesis pathways. Analysis of tissue-engineered models of the uterine wall will enable deeper investigation of molecular and biomechanical aspects of early reproductive stages (e.g. the window of implantation) or developments of pathologies.

Keywords: tissue-engineering, hormonal stimuli, reproduction, multi-layer uterine model, progesterone, β-estradiol, receptive uterine model, fertility

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1233 Use of Cellulosic Fibres in Double Layer Porous Asphalt

Authors: Márcia Afonso, Marisa Dinis-Almeida, Cristina Fael

Abstract:

Climate change, namely precipitation patterns alteration, has led to extreme conditions such as floods and droughts. In turn, excessive construction has led to the waterproofing of the soil, increasing the surface runoff and decreasing the groundwater recharge capacity. The permeable pavements used in areas with low traffic lead to a decrease in the probability of floods peaks occurrence and the sediments reduction and pollutants transport, ensuring rainwater quality improvement. This study aims to evaluate the porous asphalt performance, developed in the laboratory, with addition of cellulosic fibres. One of the main objectives of cellulosic fibres use is to stop binder drainage, preventing its loss during storage and transport. Comparing to the conventional porous asphalt the cellulosic fibres addition improved the porous asphalt performance. The cellulosic fibres allowed the bitumen content increase, enabling retention and better aggregates coating and, consequently, a greater mixture durability. With this solution, it is intended to develop better practices of resilience and adaptation to the extreme climate changes and respond to the sustainability current demands, through the eco-friendly materials use. The mix design was performed for different size aggregates (with fine aggregates – PA1 and with coarse aggregates – PA2). The percentage influence of the fibres to be used was studied. It was observed that overall, the binder drainage decreases as the cellulose fibres percentage increases. It was found that the PA2 mixture obtained most binder drainage relative to PA1 mixture, irrespective of the fibres percentage used. Subsequently, the performance was evaluated through laboratory tests of indirect tensile stiffness modulus, water sensitivity, permeability and permanent deformation. The stiffness modulus for the two mixtures groups (with and without cellulosic fibres) presented very similar values between them. For the water sensitivity test it was observed that porous asphalt containing more fine aggregates are more susceptible to the water presence than mixtures with coarse aggregates. The porous asphalt with coarse aggregates have more air voids which allow water to pass easily leading to ITSR higher values. In the permeability test was observed that asphalt porous without cellulosic fibres presented had lower permeability than asphalt porous with cellulosic fibres. The resistance to permanent deformation results indicates better behaviour of porous asphalt with cellulosic fibres, verifying a bigger rut depth in porous asphalt without cellulosic fibres. In this study, it was observed that porous asphalt with bitumen higher percentages improve the performance to permanent deformation. This fact was only possible due to the bitumen retention by the cellulosic fibres.

Keywords: binder drainage, cellulosic fibres, permanent deformation, porous asphalt

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1232 Analysis of Some Solutions to Protect the Western Tombolo of Giens

Authors: Yves Lacroix, Van Van Than, Didier Léandri, Pierre Liardet

Abstract:

The tombolo of Giens is located in the town of Hyères (France). We recall the history of coastal erosion, and prominent factors affecting the evolution of the western tombolo. We then discuss the possibility of stabilizing the western tombolo. Our argumentation relies on a coupled model integrating swells, currents, water levels and sediment transport. We present the conclusions of the simulations of various scenarios, including pre-existing propositions from coastal engineering offices. We conclude that beach replenishment seems to be necessary but not sufficient for the stabilization of the beach. Breakwaters reveal effective particularly in the most exposed northern area. Some solutions fulfill conditions so as to be elected as satisfactory. We give a comparative analysis of the efficiency of 14 alternatives for the protection of the tombolo.

Keywords: breakwaters, coupled models, replenishment, silting

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1231 Modeling Sediment Yield of Jido River in the Rift Vally

Authors: Dawit Hailekrios Hailu

Abstract:

The main objective of this study is to predict the sediment yield of the Jido River Watershed. Jido River is the largest tributary and covers around 50% of the total catchment area of Lake Shala. This research is undertaken to analyze the sediment yield of the catchments, transport capacity of the streams and sediment deposition rates of Jido River, which is located in the Sub-basin of Shala Lake, Rift Valley Basin of Ethiopia. The input data were Meteorological, Hydrological, land use/land cover maps and soil maps collected from concerned government offices. The sediment yield of Jido River and sediment change of the streams discharging into the Shala Lake were modeled.

Keywords: sediment yield, watershed, simulation, calibration

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1230 Design and Numerical Study on Aerodynamics Performance for F16 Leading Edge Extension

Authors: San-Yih Lin, Hsien-Hao Teng

Abstract:

In this research, we use commercial software, ANSYS CFX, to carry on the simulation the F16 aerodynamics performance flow field. The flight with a modified Leading Edge Extension (LEX) is proposed to increase the lift/drag ratio. The Shear Stress Transport turbulent model is used. The unstructured grid system is generated by the ICEM CFD. The prism grid around the wall surface is generated to simulate boundary layer viscosity flow field and Tetrahedron Mesh is used for the other computation domain. The lift, drag, and pitch moment are computed. The strong vortex structures upper the wing and vortex bursts under different sweep angle of LEX are investigated.

Keywords: LEX, lift/drag ratio, pitch moment, vortex burst

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1229 Ballistic Transport in One-Dimensional Random Dimer Photonic Crystals

Authors: Samira Cherid, Samir Bentata, F. Zahira Meghoufel, Sabria Terkhi, Yamina Sefir, Fatima Bendahma, Bouabdellah Bouadjemi, Ali Z. Itouni

Abstract:

In this work, we examined the propagation of light in one-dimensional systems is examined by means of the random dimer model. The introduction of defect elements, randomly in the studied system, breaks down the Anderson localization and provides a set of propagating delocalized modes at the corresponding conventional dimer resonances. However, tuning suitably the defect dimer resonance on the host ones (or vice versa), the transmission magnitudes can be enhanced providing the optimized ballistic transmission regime as an average response. Hence, ballistic optical filters can be conceived at desired wavelengths.

Keywords: photonic crystals, random dimer model, ballistic resonance, localization and transmission

Procedia PDF Downloads 528
1228 Linear Study of Electrostatic Ion Temperature Gradient Mode with Entropy Gradient Drift and Sheared Ion Flows

Authors: M. Yaqub Khan, Usman Shabbir

Abstract:

History of plasma reveals that continuous struggle of experimentalists and theorists are not fruitful for confinement up to now. It needs a change to bring the research through entropy. Approximately, all the quantities like number density, temperature, electrostatic potential, etc. are connected to entropy. Therefore, it is better to change the way of research. In ion temperature gradient mode with the help of Braginskii model, Boltzmannian electrons, effect of velocity shear is studied inculcating entropy in the magnetoplasma. New dispersion relation is derived for ion temperature gradient mode, and dependence on entropy gradient drift is seen. It is also seen velocity shear enhances the instability but in anomalous transport, its role is not seen significantly but entropy. This work will be helpful to the next step of tokamak and space plasmas.

Keywords: entropy, velocity shear, ion temperature gradient mode, drift

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1227 Quality Assessment of the Given First Aid on the Spot Events in the Opinion of Members of the Teams of the Medical Rescue in Warsaw in Poland

Authors: Aneta Binkowska, Artur Kamecki

Abstract:

The ability to provide first aid should be one of the basic skills of each of us. First aid by the Law on National Medical Emergency dated 8 September 2006 as amended, is a set of actions undertaken to save a person at the scene of an accident. In Poland, on the basis of Article 162 of the Criminal Code, we are obliged to provide first aid to the victim. In addition, according to a large part of society, unselfishness towards others in need of help is our moral obligation. The aim of the study was to learn the opinion of the members of Medical Rescue Teams (MRT) of the ‘Meditrans’ Provincial Ambulance and Sanitary Transport Service (PA and STS ‘Meditrans’) in Warsaw on how people react in real situations threatening life or health of the injured person. The study was conducted in the third quarter of 2015 on 335 members of medical rescue teams, including 77 W and 258 M, who provided medical services in the ‘Meditrans’ Provincial Ambulance and Sanitary Transport Service MRT in Warsaw. The research tool was an anonymous questionnaire survey of own design, which consisted of 12 questions: closed, half open and one open question. Respondents were divided into 3 age groups and by individual medical professions (doctor, paramedic, nurse). The straight majority of respondents encountered granting the first aid the event on the spot. However, the frequency of appearing in such proceedings isn’t too high. The first aid has most often been given in the street and in houses. The final audited fairly important element is the reason not to provide first aid by bystanders in the opinion of members of the medical teams. Respondents to this question, which was an open question were asked to name the reason for not taking any action while waiting for an ambulance. Over 50% of respondents could not answer. The most common answers were: fear, lack of knowledge and skills, reluctance, indifference, lack of training, lack of experience and fear that harm. Conclusion: The majority of respondents have encountered instances of first aid provision, but respondents assessed the frequency of such situations as low. Placing the victim in the recovery position is the simplest and most common form of first aid. Therefore, training should be introduced not only on CPR but also in the scope of helping persons in sudden health emergency, who do not have a sudden cardiac arrest. A statement can be formulated, as a main conclusion of the analysis, that only continuous education and in particular practical training will help people to overcome the barrier of their limitations in order to help others. Among the largest group of witnesses providing first aid are the elderly and youth, who are subjected to various forms of education related to first aid provision.

Keywords: BLS, first aid, medical rescue, resuscitation

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1226 Aptamers: A Potential Strategy for COVID-19 Treatment

Authors: Mohamad Ammar Ayass, Natalya Griko, Victor Pashkov, Wanying Cao, Kevin Zhu, Jin Zhang, Lina Abi Mosleh

Abstract:

Respiratory syndrome coronavirus 2 (SARS-CoV-2) is the causative agent for coronavirus disease 2019 (COVID-19). Early evidence pointed at the angiotensin-converting enzyme 2 (ACE-2) expressed on the epithelial cells of the lung as the main entry point of SARS-CoV-2 into the cells. The viral entry is mediated by the binding of the Receptor Binding Domain (RBD) of the spike protein that is expressed on the surface of the virus to the ACE-2 receptor. As the number of SARS-CoV-2 variants continues to increase, mutations arising in the RBD of SARS-CoV-2 may lead to the ineffectiveness of RBD targeted neutralizing antibodies. To address this limitation, the objective of this study is to develop a combination of aptamers that target different regions of the RBD, preventing the binding of the spike protein to ACE-2 receptor and subsequent viral entry and replication. A safe and innovative biomedical tool was developed to inhibit viral infection and reduce the harms of COVID-19. In the present study, DNA aptamers were developed against a recombinant trimer S protein using the Systematic Evolution of Ligands by Exponential enrichment (SELEX). Negative selection was introduced at round number 7 to select for aptamers that bind specifically to the RBD domain. A series of 9 aptamers (ADI2010, ADI2011, ADI201L, ADI203L, ADI205L, ADIR68, ADIR74, ADIR80, ADIR83) were selected and characterized with high binding affinity and specificity to the RBD of the spike protein. Aptamers (ADI25, ADI2009, ADI203L) were able to bind and pull down endogenous spike protein expressed on the surface of SARS-CoV-2 virus in COVID-19 positive patient samples and determined by liquid chromatography- tandem mass spectrometry analysis (LC-MS/MS). LC-MS/MS data confirmed that aptamers can bind to the RBD of the spike protein. Furthermore, results indicated that the combination of the 9 best aptamers inhibited the binding of the purified trimer spike protein to the ACE-2 receptor found on the surface of Vero E6 cells. In the same experiment, the combined aptamers displayed a better neutralizing effect than antibodies. The data suggests that the selected aptamers could be used in therapy to neutralize the effect of the SARS-CoV-2 virus by inhibiting the interaction between the RBD and ACE-2 receptor, preventing viral entry into target cells and therefore blocking viral replication.

Keywords: aptamer, ACE-2 receptor, binding inhibitor, COVID-19, spike protein, SARS-CoV-2, treatment

Procedia PDF Downloads 183