Search results for: excitation mode
759 2,7-Diazaindole as a Photophysical Probe for Excited State Hydrogen/Proton Transfer
Authors: Simran Baweja, Bhavika Kalal, Surajit Maity
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Photoinduced tautomerization reactions have been the centre of attention among the scientific community over the past several decades because of their significance in various biological systems. 7-azaindole (7AI) is considered a model system for DNA base pairing and to understand the role of such tautomerization reactions in mutations. To the best of our knowledge, extensive studies have been carried out on 7-azaindole and its solvent clusters exhibiting proton/ hydrogen transfer in both solution as well as gas phases. Derivatives of the above molecule, like 2,7- and 2,6-diazaindoles are proposed to have even better photophysical properties due to the presence of -aza group on the 2nd position. However, there are studies in the solution phase that suggest the relevance of these molecules, but there are no experimental studies reported in the gas phase yet. In our current investigation, we present the first gas phase spectroscopic data of 2,7-diazaindole (2,7-DAI) and its solvent cluster (2,7-DAI-H2O). In this, we have employed state-of-the-art laser spectroscopic methods such as fluorescence excitation (LIF), dispersed fluorescence (DF), resonant two-photon ionization-time of flight mass spectrometry (2C-R2PI), photoionization efficiency spectroscopy (PIE), IR-UV double resonance spectroscopy, i.e., fluorescence-dip infrared spectroscopy (FDIR) and resonant ion-dip infrared spectroscopy (IDIR) to understand the electronic structure of the molecule. The origin band corresponding to the S1 ← S0 transition of the bare 2,7-DAI is found to be positioned at 33910 cm-1, whereas the origin band corresponding to S1 ← S0 transition of the 2,7-DAI-H2O is positioned at 33074 cm-1. The red-shifted transition in the case of solvent cluster suggests the enhanced feasibility of excited state hydrogen/ proton transfer. The ionization potential for the 2,7-DAI molecule is found to be 8.92 eV which is significantly higher than the previously reported 7AI (8.11 eV) molecule, making it a comparatively complex molecule to study. The ionization potential is reduced by 0.14 eV in the case of 2,7-DAI-H2O (8.78 eV) cluster compared to that of 2,7-DAI. Moreover, on comparison with the available literature values of 7AI, we found the origin band of 2,7-DAI and 2,7-DAI-H2O to be red-shifted by -729 and -280 cm-1 respectively. The ground and excited state N-H stretching frequencies of the 27DAI molecule were determined using fluorescence-dip infrared spectra (FDIR) and resonant ion dip infrared spectroscopy (IDIR), obtained at 3523 and 3467 cm-1, respectively. The lower value of vNH in the electronically excited state of 27DAI implies the higher acidity of the group compared to the ground state. Moreover, we have done extensive computational analysis, which suggests that the energy barrier in the excited state reduces significantly as we increase the number of catalytic solvent molecules (S= H2O, NH3) as well as the polarity of solvent molecules. We found that the ammonia molecule is a better candidate for hydrogen transfer compared to water because of its higher gas-phase basicity. Further studies are underway to understand the excited state dynamics and photochemistry of such N-rich chromophores.Keywords: excited state hydrogen transfer, supersonic expansion, gas phase spectroscopy, IR-UV double resonance spectroscopy, laser induced fluorescence, photoionization efficiency spectroscopy
Procedia PDF Downloads 75758 Quantifying the Protein-Protein Interaction between the Ion-Channel-Forming Colicin A and the Tol Proteins by Potassium Efflux in E. coli Cells
Authors: Fadilah Aleanizy
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Colicins are a family of bacterial toxins that kill Escherichia coli and other closely related species. The mode of action of colicins involves binding to an outer membrane receptor and translocation across the cell envelope, leading to cytotoxicity through specific targets. The mechanism of colicin cytotoxicity includes a non-specific endonuclease activity or depolarization of the cytoplasmic membrane by pore-forming activity. For Group A colicins, translocation requires an interaction between the N-terminal domain of the colicin and a series of membrane- bound and periplasmic proteins known as the Tol system (TolB, TolR, TolA, TolQ, and Pal and the active domain must be translocated through the outer membranes. Protein-protein interactions are intrinsic to virtually every cellular process. The transient protein-protein interactions of the colicin include the interaction with much more complicated assemblies during colicin translocation across the cellular membrane to its target. The potassium release assay detects variation in the K+ content of bacterial cells (K+in). This assays is used to measure the effect of pore-forming colicins such as ColA on an indicator organism by measuring the changes of the K+ concentration in the external medium (K+out ) that are caused by cell killing with a K+ selective electrode. One of the goals of this work is to employ a quantifiable in-vivo method to spot which Tol protein are more implicated in the interaction with colicin A as it is translocated to its target.Keywords: K+ efflux, Colicin A, Tol-proteins, E. coli
Procedia PDF Downloads 409757 Effect of Temperature on Pervaporation Performance of Ag-Poly Vinyl Alcohol Nanocomposite Membranes
Authors: Asmaa Selim, Peter Mizsey
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Bio-ethanol is considered of higher potential as a green renewable energy source owing to its environmental benefits and its high efficiency. In the present study, silver nanoparticles were in-situ generated in a poly (vinyl alcohol) in order to improve its potentials for pervaporation of ethanol-water mixture using solution-casting. Effect of silver content on the pervaporation separation index and the enrichment factor of the membrane at 15 percentage mass water at 40ᵒC was reported. Pervaporation data for nanocomposite membranes showed around 100% increase in the water permeance values while the intrinsic selectivity decreased. The water permeances of origin crosslinked PVA membrane, and the 2.5% silver loaded PVA membrane are 26.65 and 70.45 (g/m².kPa.h) respectively. The values of total flux and water flux are closed to each other, indicating that membranes could be effectively used to break the azeotropic point of ethanol-water. Effect of temperature on the pervaporation performance, permeation parameter and diffusion coefficient of both water and ethanol was discussed. The negative heat of sorption ∆Hs values calculated on the basis of the estimated Arrhenius activation energy values indicating that the sorption process was controlled by Langmuir’s mode. The overall results showed that the membrane containing 0.5 mass percentage of Ag salt exhibited excellent PV performance.Keywords: bio-ethanol, diffusion coefficient, nanocomposite, pervaporation, poly (vinyl alcohol), silver nanoparticles
Procedia PDF Downloads 170756 Material and Parameter Analysis of the PolyJet Process for Mold Making Using Design of Experiments
Authors: A. Kampker, K. Kreisköther, C. Reinders
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Since additive manufacturing technologies constantly advance, the use of this technology in mold making seems reasonable. Many manufacturers of additive manufacturing machines, however, do not offer any suggestions on how to parameterize the machine to achieve optimal results for mold making. The purpose of this research is to determine the interdependencies of different materials and parameters within the PolyJet process by using design of experiments (DoE), to additively manufacture molds, e.g. for thermoforming and injection molding applications. Therefore, the general requirements of thermoforming molds, such as heat resistance, surface quality and hardness, have been identified. Then, different materials and parameters of the PolyJet process, such as the orientation of the printed part, the layer thickness, the printing mode (matte or glossy), the distance between printed parts and the scaling of parts, have been examined. The multifactorial analysis covers the following properties of the printed samples: Tensile strength, tensile modulus, bending strength, elongation at break, surface quality, heat deflection temperature and surface hardness. The key objective of this research is that by joining the results from the DoE with the requirements of the mold making, optimal and tailored molds can be additively manufactured with the PolyJet process. These additively manufactured molds can then be used in prototyping processes, in process testing and in small to medium batch production.Keywords: additive manufacturing, design of experiments, mold making, PolyJet, 3D-Printing
Procedia PDF Downloads 255755 Optical and Structural Characterization of Rare Earth Doped Phosphate Glasses
Authors: Zélia Maria Da Costa Ludwig, Maria José Valenzuela Bell, Geraldo Henriques Da Silva, Thales Alves Faraco, Victor Rocha Da Silva, Daniel Rotmeister Teixeira, Vírgilio De Carvalho Dos Anjos, Valdemir Ludwig
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Advances in telecommunications grow with the development of optical amplifiers based on rare earth ions. The focus has been concentrated in silicate glasses although their amplified spontaneous emission is limited to a few tens of nanometers (~ 40nm). Recently, phosphate glasses have received great attention due to their potential application in optical data transmission, detection, sensors and laser detector, waveguide and optical fibers, besides its excellent physical properties such as high thermal expansion coefficients and low melting temperature. Compared with the silica glasses, phosphate glasses provide different optical properties such as, large transmission window of infrared, and good density. Research on the improvement of physical and chemical durability of phosphate glass by addition of heavy metals oxides in P2O5 has been performed. The addition of Na2O further improves the solubility of rare earths, while increasing the Al2O3 links in the P2O5 tetrahedral results in increased durability and aqueous transition temperature and a decrease of the coefficient of thermal expansion. This work describes the structural and spectroscopic characterization of a phosphate glass matrix doped with different Er (Erbium) concentrations. The phosphate glasses containing Er3+ ions have been prepared by melt technique. A study of the optical absorption, luminescence and lifetime was conducted in order to characterize the infrared emission of Er3+ ions at 1540 nm, due to the radiative transition 4I13/2 → 4I15/2. Our results indicate that the present glass is a quite good matrix for Er3+ ions, and the quantum efficiency of the 1540 nm emission was high. A quenching mechanism for the mentioned luminescence was not observed up to 2,0 mol% of Er concentration. The Judd-Ofelt parameters, radiative lifetime and quantum efficiency have been determined in order to evaluate the potential of Er3+ ions in new phosphate glass. The parameters follow the trend as Ω2 > Ω4 > Ω6. It is well known that the parameter Ω2 is an indication of the dominant covalent nature and/or structural changes in the vicinity of the ion (short range effects), while Ω4 and Ω6 intensity parameters are long range parameters that can be related to the bulk properties such as viscosity and rigidity of the glass. From the PL measurements, no red or green upconversion was measured when pumping the samples with laser excitation at 980 nm. As future prospects: Synthesize this glass system with silver in order to determine the influence of silver nanoparticles on the Er3+ ions.Keywords: phosphate glass, erbium, luminescence, glass system
Procedia PDF Downloads 510754 An Ideational Grammatical Metaphor of Narrative History in Chinua Achebe's 'There Was a Country'
Authors: Muhammed-Badar Salihu Jibrin, Chibabi Makedono Darlington
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This paper studied Ideational Grammatical Metaphor (IGM) of Narrative History in Chinua Achebe’s There Was a Country. It started with a narrative historical style as a recent genre out of the conventional historical writings. In order to explore the linguistic phenomenon using a particular lexico-grammatical tool of IGM, the theoretical background was examined based on Hallidayan Systemic Functional Linguistics. Furthermore, the study considered the possibility of applying IGM to the Part 4 of Achebe’s historical text with recourse to the concept of congruence in IGM and research questions before formulating a working methodology. The analysis of Achebe’s memoir was, thus, presented in tabular forms to account for the quantitative content analysis with qualitative research technique, as well as the metaphorical and congruent wording through nominalization and process types with samples. The frequencies and percentage were given appropriately with respect to each subheadings of the text. To this end, the findings showed that material and relational types indicated dominance. The discussion and implications were that the findings confirmed earlier study by MAK Halliday and C.I.M.I.M. Matthiessen’s suggestion that IGM should show dominance of material type process. The implication is that IGM can be an effective tool for the analysis of a narrative historical text. In conclusion, it was observed that IGM does not only carry grammatical function but also an ideological role in shaping the historical discourse within the narrative mode between writers and readers.Keywords: ideational grammatical metaphor, nominalization, narrative history, memoire, dominance
Procedia PDF Downloads 220753 Production of Nitric Oxide by Thienopyrimidine TP053
Authors: Elena G. Salina, Laurent R. Chiarelli, Maria R. Pasca, Vadim A. Makarov
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Tuberculosis is one of the most challenging threats to human health, confronted by the problem of drug resistance. Evidently, new drugs for tuberculosis are urgently needed. Thienopyrimidine TP053 is one of the most promising new antitubercular prodrugs. Mycothiol-dependent reductase Mrx2, encoded by rv2466c, is known to be a TP053 activator; however, the precise mode of action of this compound remained unclear. Being highly active against both replicating and non-replicating tuberculosis bacilli, TP053 also revealed dose-escalating activity for M. tuberculosis-infected murine macrophages. The chemical structure of TP053 is characterized by the presence of NO₂ group which was suggested to be responsible for the toxic effects of the activated compound. Reduction of a nitroaromatic moiety of TP53 by Mrx2 was hypothesized to result in NO release. Analysis of the products of enzymatic activation of TP053 by Mrx2 by the Greiss reagent clearly demonstrated production of nitric oxide in a time-dependent manner. Mass-spectra of cell lysates of TP-treated M. tuberculosis bacilli demonstrated the transformation of TP053 to its non-active metabolite with Mw=261 that corresponds NO release. The mechanism of NO toxicity for bacteria includes DNA damage and degradation of iron-sulfur centers, especially under oxygen depletion. Thus, TP-053 drug-like scaffold is prospective for further development of novel anti-TB drug. This work was financially supported by the Russian Foundation for Basic Research (Grant 17-04-00342).Keywords: drug discovery, M. tuberculosis, nitric oxide, NO donors
Procedia PDF Downloads 153752 Research on the Effect of Accelerated Aging Illumination Mode on Bifacial Solar Modules
Authors: T. H. Huang, C. L. Fern, Y. K. Tseng
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The design and reliability of solar photovoltaic modules are crucial to the development of solar energy, and efforts are still being made to extend the life of photovoltaic modules to improve their efficiency because natural aging is time-consuming and does not provide manufacturers and investors with timely information, accelerated aging is currently the best way to estimate the life of photovoltaic modules. Bifacial solar cells not only absorb light from the front side but also absorb light reflected from the ground on the back side, surpassing the performance of single-sided solar cells. Due to the asymmetry of the two sides of the light, in addition to the difference in photovoltaic conversion efficiency, there will also be differences in heat distribution, which will affect the electrical properties and material structure of the bifacial solar cell itself. In this study, there are two types of experimental samples: packaged and unpackaged and then irradiated with UVC light sources and halogen lamps for accelerated aging, as well as a control group without aging. After two weeks of accelerated aging, the bifacial solar cells were visual observation, and infrared thermal images were taken; then, the samples were subjected to IV measurement, and samples were taken for SEM, Raman, and XRD analyses in order to identify the defects that lead to failure and chemical changes, as well as to analyze the reasons for the degradation of their characteristics. From the results of the analysis, it is found that aging will cause carbonization of the polymer material on the surface of bifacial solar cells, and the crystal structure will be affected.Keywords: bifacial solar cell, accelerated aging, temperature, characterization, electrical measurement
Procedia PDF Downloads 112751 X-Ray Dosimetry by a Low-Cost Current Mode Ion Chamber
Authors: Ava Zarif Sanayei, Mustafa Farjad-Fard, Mohammad-Reza Mohammadian-Behbahani, Leyli Ebrahimi, Sedigheh Sina
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The fabrication and testing of a low-cost air-filled ion chamber for X-ray dosimetry is studied. The chamber is made of a metal cylinder, a central wire, a BC517 Darlington transistor, a 9V DC battery, and a voltmeter in order to have a cost-effective means to measure the dose. The output current of the dosimeter is amplified by the transistor and then fed to the large internal resistance of the voltmeter, producing a readable voltage signal. The dose-response linearity of the ion chamber is evaluated for different exposure scenarios by the X-ray tube. kVp values 70, 90, and 120, and mAs up to 20 are considered. In all experiments, a solid-state dosimeter (Solidose 400, Elimpex Medizintechnik) is used as a reference device for chamber calibration. Each case of exposure is repeated three times, the voltmeter and Solidose readings are recorded, and the mean and standard deviation values are calculated. Then, the calibration curve, derived by plotting voltmeter readings against Solidose readings, provided a linear fit result for all tube kVps of 70, 90, and 120. A 99, 98, and 100% linear relationship, respectively, for kVp values 70, 90, and 120 are demonstrated. The study shows the feasibility of achieving acceptable dose measurements with a simplified setup. Further enhancements to the proposed setup include solutions for limiting the leakage current, optimizing chamber dimensions, utilizing electronic microcontrollers for dedicated data readout, and minimizing the impact of stray electromagnetic fields on the system.Keywords: dosimetry, ion chamber, radiation detection, X-ray
Procedia PDF Downloads 77750 A Research on the Effect of Soil-Structure Interaction on the Dynamic Response of Symmetrical Reinforced Concrete Buildings
Authors: Adinew Gebremeskel Tizazu
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The effect of soil-structure interaction on the dynamic response of reinforced concrete buildings of regular and symmetrical geometry are considered in this study. The structures are presumed to be generally embedded in a homogenous soil formation underlain by very stiff material or bedrock. The structure-foundation–soil system is excited at the base by an earthquake ground motion. The superstructure is idealized as a system with lumped masses concentrated at the floor levels, and coupled with the substructure. The substructure system, which comprises of the foundation and soil, is represented, and replaced by springs and dashpots. Frequency-dependent impedances of the foundation system are incorporated in the discrete model in terms of the springs and dashpots coefficients. The excitation applied to the model is field ground motions of actual earthquake records. Modal superposition principle is employed to transform the equations of motion in geometrical coordinates to modal coordinates. However, the modal equations remain coupled with respect to damping terms due to the difference in damping mechanisms of the superstructure and the soil. Hence, proportional damping for the coupled structural system may not be assumed. An iterative approach is adopted and programmed to solve the system of coupled equations of motion in modal coordinates to obtain the displacement responses of the system. Parametric studies for responses of building structures with regular and symmetric plans of different structural properties and heights are made for fixed and flexible base conditions, for different soil conditions encountered in Addis Ababa. The displacement, base shear and base overturning moments are used in the comparison of different types of structures for various foundation embedment depths, site conditions and height of structures. These values are compared against those of fixed base structure. The study shows that the flexible base structures, generally exhibit different responses from those structures with fixed base. Basically, the natural circular frequencies, the base shears and the inter-story displacements for the flexible base are less than those of the fixed base structures. This trend is particularly evident when the flexible soil has large thickness. In contrast, the trend becomes less predictable, when the thickness of the flexible soil decreases. Moreover, in the latter case, the iteration undulates significantly making the prediction difficult. This is attributed to the highly jagged nature of the impedance functions of frequencies for such formations. In this case, it is difficult to conclude whether the conventional fixed-base approach yields conservative design forces, as is the case for soil formations of large thickness.Keywords: effect of soil structure, dynamic response corroborated, the modal superposition principle, parametric studies
Procedia PDF Downloads 32749 Study on Electromagnetic Plasma Acceleration Using Rotating Magnetic Field Scheme
Authors: Takeru Furuawa, Kohei Takizawa, Daisuke Kuwahara, Shunjiro Shinohara
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In the field of a space propulsion, an electric propulsion system has been developed because its fuel efficiency is much higher than a conventional chemical one. However, the practical electric propulsion systems, e.g., an ion engine, have a problem of short lifetime due to a damage of generation and acceleration electrodes of the plasma. A helicon plasma thruster is proposed as a long-lifetime electric thruster which has non-direct contact electrodes. In this system, both generation and acceleration methods of a dense plasma are executed by antennas from the outside of a discharge tube. Development of the helicon plasma thruster has been conducting under the Helicon Electrodeless Advanced Thruster (HEAT) project. Our helicon plasma thruster has two important processes. First, we generate a dense source plasma using a helicon wave with an excitation frequency between an ion and an electron cyclotron frequencies, fci and fce, respectively, applied from the outside of a discharge using a radio frequency (RF) antenna. The helicon plasma source can provide a high-density (~1019 m-3), a high-ionization ratio (up to several tens of percent), and a high particle generation efficiency. Second, in order to achieve high thrust and specific impulse, we accelerate the dense plasma by the axial Lorentz force fz using the product of the induced azimuthal current jθ and the static radial magnetic field Br, shown as fz = jθ × Br. The HEAT project has proposed several kinds of electrodeless acceleration schemes, and in our particular case, a Rotating Magnetic Field (RMF) method has been extensively studied. The RMF scheme was originally developed as a concept to maintain the Field Reversed Configuration (FRC) in a magnetically confined fusion research. Here, RMF coils are expected to generate jθ due to a nonlinear effect shown below. First, the rotating magnetic field Bω is generated by two pairs of RMF coils with AC currents, which have a phase difference of 90 degrees between the pairs. Due to the Faraday’s law, an axial electric field is induced. Second, an axial current is generated by the effects of an electron-ion and an electron-neutral collisions through the Ohm’s law. Third, the azimuthal electric field is generated by the nonlinear term, and the retarding torque generated by the collision effects again. Then, azimuthal current jθ is generated as jθ = - nₑ er ∙ 2π fRMF. Finally, the axial Lorentz force fz for plasma acceleration is generated. Here, jθ is proportional to nₑ and frequency of RMF coil current fRMF, when Bω is fully penetrated into the plasma. Our previous study has achieved 19 % increase of ion velocity using the 5 MHz and 50 A of the RMF coil power supply. In this presentation, we will show the improvement of the ion velocity using the lower frequency and higher current supplied by RMF power supply. In conclusion, helicon high-density plasma production and electromagnetic acceleration by the RMF scheme with a concept of electrodeless condition have been successfully executed.Keywords: electric propulsion, electrodeless thruster, helicon plasma, rotating magnetic field
Procedia PDF Downloads 261748 Effect of PMMA Shield on the Patient Dose Equivalent from Photoneutrons Produced by High Energy Medical Linacs
Authors: Seyed Mehdi Hashemi, Gholamreza Raisali, Mehran Taheri
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One of the important problems of using high energy linacs at IMRT is the production of photoneutrons. Besides the clinically useful photon beams, high-energy photon beams from medical linacs produce secondary neutrons. These photoneutrons increase the patient dose and may cause secondary malignancies. The effect of the shield on the reduction of photoneutron dose equivalent produced by a high energy medical linac at the patient plane is investigated in this study. To determine the photoneutron dose equivalent received to the patient a Varian linac working at 18 MV photon mode investigated. Photoneutron dose equivalent measured with Polycarbonate films of 0.25 mm thick. PC films placed at distances of 0, 10, 20, and 50 cm from the center of X-ray field on the patient couch. The results show that by increasing the distance from the center of the X-ray beam towards the periphery, the photoneutron dose equivalent decreases rapidly for both open and shielded fields and that by inserting the shield in the path of the X-ray beam, the photoneutron dose equivalent was decreased obviously compared to open field. Results show the shield, significantly reduces photoneutron dose equivalent to the patient. Results can be readily generalized to other models of medical linacs. It may be concluded that using this kind of shield can help more safe, inexpensive and efficient employment of high energy linacs in radiotherapy and IMRT.Keywords: photoneutron, Linac, PMMA shield, equivalent dose
Procedia PDF Downloads 493747 Preparation and Characterization of Phosphate-Nickel-Titanium Composite Coating Obtained by Sol Gel Process for Corrosion Protection
Authors: Khalidou Ba, Abdelkrim Chahine, Mohamed Ebn Touhami
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A strong industrial interest is focused on the development of coatings for anticorrosion protection. In this context, phosphate composite materials are expanding strongly due to their chemical characteristics and their interesting physicochemical properties. Sol-gel coatings offer high homogeneity and purity that may lead to obtain coating presenting good adhesion to metal surface. The goal behind this work is to develop efficient coatings for corrosion protection of steel to extend its life. In this context, a sol gel process allowing to obtain thin film coatings on carbon steel with high resistance to corrosion has been developed. The optimization of several experimental parameters such as the hydrolysis time, the temperature, the coating technique, the molar ratio between precursors, the number of layers and the drying mode has been realized in order to obtain a coating showing the best anti-corrosion properties. The effect of these parameters on the microstructure and anticorrosion performance of the films sol gel coating has been investigated using different characterization methods (FTIR, XRD, Raman, XPS, SEM, Profilometer, Salt Spray Test, etc.). An optimized coating presenting good adhesion and very stable anticorrosion properties in salt spray test, which consists of a corrosive attack accelerated by an artificial salt spray consisting of a solution of 5% NaCl, pH neutral, under precise conditions of temperature (35 °C) and pressure has been obtained.Keywords: sol gel, coating, corrosion, XPS
Procedia PDF Downloads 128746 Study on Environmental Capacity System of the Aged Care Villages Influenced by Tourists
Authors: Yuan Fang, Wang-Ming Li, Yi-Chen Ruan
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Rural healthy old-age care for urban elderly who go to surrounding villages on vacation is a new mode of old-age care in developed coastal areas of China. Such villages that receive urban elderly can be called old-caring villages. Due to the popularity of healthy old-age care in rural areas, more and more urban elderly people participate in the ranks of rural old-age care, resulting in excessive number of tourists in some old-caring villages, exceeding the carrying capacity of the village. Excessive passenger flow may damage the ecological environment, social environment, and facilities environment of the village, and even affect the development potential of the village pension industry. On the basis of on-site investigation and questionnaire survey, this paper summarizes the willingness and behavioral characteristics of the urban elderly population and finds that it will have a certain impact on the old-caring villages in the process of pension vacation in the aspects of ecology, construction, society, and economy. According to the influence of tourists, the paper constructs a system of capacity restriction factors of the old-caring villages, which includes four types: ecological environment capacity, policy environment capacity, perceived congestion capacity, and village service capacity, and fourteen specific indicators. It will provide a theoretical basis for reasonable control of the development scale of the old-caring villages.Keywords: old-caring villages, restriction factors system, tourists' influence, environmental capacity
Procedia PDF Downloads 148745 Performance Enhancement of Hybrid Racing Car by Design Optimization
Authors: Tarang Varmora, Krupa Shah, Karan Patel
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Environmental pollution and shortage of conventional fuel are the main concerns in the transportation sector. Most of the vehicles use an internal combustion engine (ICE), powered by gasoline fuels. This results into emission of toxic gases. Hybrid electric vehicle (HEV) powered by electric machine and ICE is capable of reducing emission of toxic gases and fuel consumption. However to build HEV, it is required to accommodate motor and batteries in the vehicle along with engine and fuel tank. Thus, overall weight of the vehicle increases. To improve the fuel economy and acceleration, the weight of the HEV can be minimized. In this paper, the design methodology to reduce the weight of the hybrid racing car is proposed. To this end, the chassis design is optimized. Further, attempt is made to obtain the maximum strength with minimum material weight. The best configuration out of the three main configurations such as series, parallel and the dual-mode (series-parallel) is chosen. Moreover, the most suitable type of motor, battery, braking system, steering system and suspension system are identified. The racing car is designed and analyzed in the simulating software. The safety of the vehicle is assured by performing static and dynamic analysis on the chassis frame. From the results, it is observed that, the weight of the racing car is reduced by 11 % without compromising on safety and cost. It is believed that the proposed design and specifications can be implemented practically for manufacturing hybrid racing car.Keywords: design optimization, hybrid racing car, simulation, vehicle, weight reduction
Procedia PDF Downloads 294744 Study of Rayleigh-Bénard-Brinkman Convection Using LTNE Model and Coupled, Real Ginzburg-Landau Equations
Authors: P. G. Siddheshwar, R. K. Vanishree, C. Kanchana
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A local nonlinear stability analysis using a eight-mode expansion is performed in arriving at the coupled amplitude equations for Rayleigh-Bénard-Brinkman convection (RBBC) in the presence of LTNE effects. Streamlines and isotherms are obtained in the two-dimensional unsteady finite-amplitude convection regime. The parameters’ influence on heat transport is found to be more pronounced at small time than at long times. Results of the Rayleigh-Bénard convection is obtained as a particular case of the present study. Additional modes are shown not to significantly influence the heat transport thus leading us to infer that five minimal modes are sufficient to make a study of RBBC. The present problem that uses rolls as a pattern of manifestation of instability is a needed first step in the direction of making a very general non-local study of two-dimensional unsteady convection. The results may be useful in determining the preferred range of parameters’ values while making rheometric measurements in fluids to ascertain fluid properties such as viscosity. The results of LTE are obtained as a limiting case of the results of LTNE obtained in the paper.Keywords: coupled Ginzburg–Landau model, local thermal non-equilibrium (LTNE), local thermal equilibrium (LTE), Rayleigh–Bénard-Brinkman convection
Procedia PDF Downloads 237743 Gold-Mediated Modification of Apoferritin Surface with Targeting Antibodies
Authors: Simona Dostalova, Pavel Kopel, Marketa Vaculovicova, Vojtech Adam, Rene Kizek
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Protein apoferritin seems to be a very promising structure for use as a nanocarrier. It is prepared from intracellular ferritin protein naturally found in most organisms. The role of ferritin proteins is to store and transport ferrous ions. Apoferritin is a hollow protein cage without ferrous ions that can be prepared from ferritin by reduction with thioglycolic acid or dithionite. The structure of apoferritin is composed of 24 protein subunits, creating a sphere with 12 nm in diameter. The inner cavity has a diameter of 8 nm. The drug encapsulation process is based on the response of apoferritin structure to the pH changes of surrounding solution. In low pH, apoferritin is disassembled into individual subunits and its structure is “opened”. It can then be mixed with any desired cytotoxic drug and after adjustment of pH back to neutral the subunits are reconnected again and the drug is encapsulated within the apoferritin particles. Excess drug molecules can be removed by dialysis. The receptors for apoferritin, SCARA5 and TfR1 can be found in the membrane of both healthy and cancer cells. To enhance the specific targeting of apoferritin nanocarrier, it is possible to modify its surface with targeting moieties, such as antibodies. To ensure sterically correct complex, we used a a peptide linker based on a protein G with N-terminus affinity towards Fc region of antibodies. To connect the peptide to the surface of apoferritin, the C-terminus of peptide was made of cysteine with affinity to gold. The surface of apoferritin with encapsulated doxorubicin (ApoDox) was coated either with gold nanoparticles (ApoDox-Nano) or gold (III) chloride hydrate reduced with sodium borohydride (ApoDox-HAu). The applied amount of gold in form of gold (III) chloride hydrate was 10 times higher than in the case of gold nanoparticles. However, after removal of the excess unbound ions by electrophoretic separation, the concentration of gold on the surface of apoferritin was only 6 times higher for ApoDox-HAu in comparison with ApoDox-Nano. Moreover, the reduction with sodium borohydride caused a loss of doxorubicin fluorescent properties (excitation maximum at 480 nm with emission maximum at 600 nm) and thus its biological activity. Fluorescent properties of ApoDox-Nano were similar to the unmodified ApoDox, therefore it was more suited for the intended use. To evaluate the specificity of apoferritin modified with antibodies, we used ELISA-like method with the surface of microtitration plate wells coated by the antigen (goat anti-human IgG antibodies). To these wells, we applied ApoDox without targeting antibodies and ApoDox-Nano modified with targeting antibodies (human IgG antibodies). The amount of unmodified ApoDox on antigen after incubation and subsequent rinsing with water was 5 times lower than in the case of ApoDox-Nano modified with targeting antibodies. The modification of non-gold ApoDox with antibodies caused no change in its targeting properties. It can therefore be concluded that the demonstrated procedure allows us to create nanocarrier with enhanced targeting properties, suitable for nanomedicine.Keywords: apoferritin, doxorubicin, nanocarrier, targeting antibodies
Procedia PDF Downloads 389742 Estimation of Time Loss and Costs of Traffic Congestion: The Contingent Valuation Method
Authors: Amira Mabrouk, Chokri Abdennadher
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The reduction of road congestion which is inherent to the use of vehicles is an obvious priority to public authority. Therefore, assessing the willingness to pay of an individual in order to save trip-time is akin to estimating the change in price which was the result of setting up a new transport policy to increase the networks fluidity and improving the level of social welfare. This study holds an innovative perspective. In fact, it initiates an economic calculation that has the objective of giving an estimation of the monetized time value during the trips made in Sfax. This research is founded on a double-objective approach. The aim of this study is to i) give an estimation of the monetized value of time; an hour dedicated to trips, ii) determine whether or not the consumer considers the environmental variables to be significant, iii) analyze the impact of applying a public management of the congestion via imposing taxation of city tolls on urban dwellers. This article is built upon a rich field survey led in the city of Sfax. With the use of the contingent valuation method, we analyze the “declared time preferences” of 450 drivers during rush hours. Based on the fond consideration of attributed bias of the applied method, we bring to light the delicacy of this approach with regards to the revelation mode and the interrogative techniques by following the NOAA panel recommendations bearing the exception of the valorization point and other similar studies about the estimation of transportation externality.Keywords: willingness to pay, contingent valuation, time value, city toll
Procedia PDF Downloads 434741 Effect of Site Amplification on Seismic Safety Evaluation of Flyover Pier
Authors: Mohammad Raihan Mukhlis, M. Abdur Rahman Bhuiyan
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Bangladesh is a developing country in which a lot of multi-span simply/continuous supported flyovers are being constructed in its major cities. Being situated in a seismically active region, seismic safety evaluation of flyovers is essential for seismic risk reduction. Effects of site amplification on seismic safety evaluation of flyover piers are the main concern of this study. In this regard, failure mode, lateral strength and displacement ductility of piers of a typical multi-span simply supported flyover have been evaluated by Japan Road Association (JRA) recommended guidelines, with and without considering site amplification. Ultimate flexural strengths of piers have been computed using the pushover analysis results. Shear capacity of piers has been calculated using the guidelines of JRA. Lateral strengths have been determined depending on the failure modes of the piers. Displacement ductility of piers has been computed using yield and ultimate displacements of the piers obtained from the pushover analysis results. Selected earthquake time history is used in seismic safety evaluation of the flyover piers. Finally, the ductility design method is used to conduct the seismic safety evaluation of the piers with and without considering site amplification. From the numerical results, it has been revealed that the effects of site amplification on seismic safety evaluation of bridge structures should be carefully taken into account.Keywords: displacement ductility, flyover pier, lateral strength, safety evaluation, site amplification
Procedia PDF Downloads 170740 Disruption of Cancer Cell Proliferation by Magnetic Field
Authors: Ming Ze Kao
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Static magnetic fields (SMF) are widely used in several medical applications, especially in diagnosis of tumors. However, biological effects of the SMFs on modulating cell physiology through the Lorentz force, which is highly frequency and magnitude dependent, remain to be elucidated. Specific patterns from SMFs of static MF, delivered by means of Halbach array magnets with a gradient increment of 6.857mT/mm from center to border, were found to have profound inhibitory effect on the growth rate of human cell line derived from Nasopharyngeal carcinoma patients. The SMFs, which were shown to be noncontact, selectively impact rapid dividing cells while quiescent cells stay intact. The phenomenon acts in two modes: the arrest of cell proliferation in the G2/M phase and destruction of cell mitosis in cell division. First mode is manifested by impacting the proper formation of mitotic spindle, whereas the second results in disintegration of the cancer cell. Both modes are demonstrated when SMF was applied for 24 hours to cancer cells, the results revealed that metaphase arrest during mitosis due to activation of DNA damage response (DDR), resulting in high expression of ATM-NBS1-CHEK signaling pathways and higher G2/M phase ratio compared with control group. Here, experimental data suggest that the SMFs cause activation of cell cycle checkpoints, which implies the MFs as a potential therapeutic modality as a sensitizer for radiotherapy or chemotherapy.Keywords: static magnetic field, DNA damage response, Halbach array, magnetic therapy
Procedia PDF Downloads 114739 A Biomimetic Structural Form: Developing a Paradigm to Attain Vital Sustainability in Tall Architecture
Authors: Osama Al-Sehail
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This paper argues for sustainability as a necessity in the evolution of tall architecture. It provides a different mode for dealing with sustainability in tall architecture, taking into consideration the speciality of its typology. To this end, the article develops a Biomimetic Structural Form as a paradigm to attain Vital Sustainability. A Biomimetic Structural Form, which is derived from the amalgamation of biomimicry as an approach for sustainability defining nature as source of knowledge and inspiration in solving humans’ problems and a Structural Form as a catalyst for evolving tall architecture, is a dynamic paradigm emerging from a conceptualizing and morphological process. A Biomimetic Structural Form is a flow system whose different forces and functions tend to be “better”, more "fit", to “survive”, and to be efficient. Through geometry and function—the two aspects of knowledge extracted from nature—the attributes of the Biomimetic Structural Form are formulated. Vital Sustainability is the survival level of sustainability in natural systems through which a system enhances the performance of its internal working and its interaction with the external environment. A Biomimetic Structural Form, in this context, is a medium for evolving tall architecture to emulate natural models in their ways of coexistence with the environment. As an integral part of this article, the sustainable super tall building 3Ts is discussed as a case study of applying Biomimetic Structural Form.Keywords: biomimicry, design in nature, high-rise buildings, sustainability, structural form, tall architecture, vital sustainability
Procedia PDF Downloads 312738 Dynamic Risk Identification Using Fuzzy Failure Mode Effect Analysis in Fabric Process Industries: A Research Article as Management Perspective
Authors: A. Sivakumar, S. S. Darun Prakash, P. Navaneethakrishnan
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In and around Erode District, it is estimated that more than 1250 chemical and allied textile processing fabric industries are affected, partially closed and shut off for various reasons such as poor management, poor supplier performance, lack of planning for productivity, fluctuation of output, poor investment, waste analysis, labor problems, capital/labor ratio, accumulation of stocks, poor maintenance of resources, deficiencies in the quality of fabric, low capacity utilization, age of plant and equipment, high investment and input but low throughput, poor research and development, lack of energy, workers’ fear of loss of jobs, work force mix and work ethic. The main objective of this work is to analyze the existing conditions in textile fabric sector, validate the break even of Total Productivity (TP), analyze, design and implement fuzzy sets and mathematical programming for improvement of productivity and quality dimensions in the fabric processing industry. It needs to be compatible with the reality of textile and fabric processing industries. The highly risk events from productivity and quality dimension were found by fuzzy systems and results are wrapped up among the textile fabric processing industry.Keywords: break even point, fuzzy crisp data, fuzzy sets, productivity, productivity cycle, total productive maintenance
Procedia PDF Downloads 338737 A Fast Calculation Approach for Position Identification in a Distance Space
Authors: Dawei Cai, Yuya Tokuda
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The market of localization based service (LBS) is expanding. The acquisition of physical location is the fundamental basis for LBS. GPS, the de facto standard for outdoor localization, does not work well in indoor environment due to the blocking of signals by walls and ceiling. To acquire high accurate localization in an indoor environment, many techniques have been developed. Triangulation approach is often used for identifying the location, but a heavy and complex computation is necessary to calculate the location of the distances between the object and several source points. This computation is also time and power consumption, and not favorable to a mobile device that needs a long action life with battery. To provide a low power consumption approach for a mobile device, this paper presents a fast calculation approach to identify the location of the object without online solving solutions to simultaneous quadratic equations. In our approach, we divide the location identification into two parts, one is offline, and other is online. In offline mode, we make a mapping process that maps the location area to distance space and find a simple formula that can be used to identify the location of the object online with very light computation. The characteristic of the approach is a good tradeoff between the accuracy and computational amount. Therefore, this approach can be used in smartphone and other mobile devices that need a long work time. To show the performance, some simulation experimental results are provided also in the paper.Keywords: indoor localization, location based service, triangulation, fast calculation, mobile device
Procedia PDF Downloads 174736 Performance Evaluation of Reinforced Concrete Framed Structure with Steel Bracing and Supplemental Energy Dissipation
Authors: Swanand Patil, Pankaj Agarwal
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In past few decades, seismic performance objectives have shifted from earthquake resistance to earthquake resilience of the structures, especially for the lifeline buildings. Features such as negligible post-earthquake damage and replaceable damaged components, makes energy dissipating systems a valid choice for a seismically resilient building. In this study, various energy dissipation devices are applied on an eight-storey moment resisting RC building model. The energy dissipating devices include both hysteresis-based and viscous type of devices. The seismic response of the building is obtained for different positioning and mechanical properties of the devices. The investigation is carried forward to the deficiently ductile RC frame also. The performance assessment is done on the basis of drift ratio, mode shapes and displacement response of the model structures. Nonlinear dynamic analysis shows largely improved displacement response. The damping devices improve displacement response more efficiently in the deficient ductile frames than that in the perfectly moment resisting frames. This finding is important considering the number of deficient buildings in India and the world. The placement and mechanical properties of the dampers prove to be a crucial part in modelling, analyzing and designing of the structures with supplemental energy dissipation.Keywords: earthquake resilient structures, lifeline buildings, retrofitting of structures, supplemental energy dissipation
Procedia PDF Downloads 345735 The Effect of Using Emg-based Luna Neurorobotics for Strengthening of Affected Side in Chronic Stroke Patients - Retrospective Study
Authors: Surbhi Kaura, Sachin Kandhari, Shahiduz Zafar
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Chronic stroke, characterized by persistent motor deficits, often necessitates comprehensive rehabilitation interventions to improve functional outcomes and mitigate long-term dependency. Luna neurorobotic devices, integrated with EMG feedback systems, provide an innovative platform for facilitating neuroplasticity and functional improvement in stroke survivors. This retrospective study aims to investigate the impact of EMG-based Luna neurorobotic interventions on the strengthening of the affected side in chronic stroke patients. In rehabilitation, active patient participation significantly activates the sensorimotor network during motor control, unlike passive movement. Stroke is a debilitating condition that, when not effectively treated, can result in significant deficits and lifelong dependency. Common issues like neglecting the use of limbs can lead to weakness in chronic stroke cases. In rehabilitation, active patient participation significantly activates the sensorimotor network during motor control, unlike passive movement. This study aims to assess how electromyographic triggering (EMG-triggered) robotic treatments affect walking, ankle muscle force after an ischemic stroke, and the coactivation of agonist and antagonist muscles, which contributes to neuroplasticity with the assistance of biofeedback using robotics. Methods: The study utilized robotic techniques based on electromyography (EMG) for daily rehabilitation in long-term stroke patients, offering feedback and monitoring progress. Each patient received one session per day for two weeks, with the intervention group undergoing 45 minutes of robot-assisted training and exercise at the hospital, while the control group performed exercises at home. Eight participants with impaired motor function and gait after stroke were involved in the study. EMG-based biofeedback exercises were administered through the LUNA neuro-robotic machine, progressing from trigger and release mode to trigger and hold, and later transitioning to dynamic mode. Assessments were conducted at baseline and after two weeks, including the Timed Up and Go (TUG) test, a 10-meter walk test (10m), Berg Balance Scale (BBG), and gait parameters like cadence, step length, upper limb strength measured by EMG threshold in microvolts, and force in Newton meters. Results: The study utilized a scale to assess motor strength and balance, illustrating the benefits of EMG-biofeedback following LUNA robotic therapy. In the analysis of the left hemiparetic group, an increase in strength post-rehabilitation was observed. The pre-TUG mean value was 72.4, which decreased to 42.4 ± 0.03880133 seconds post-rehabilitation, with a significant difference indicated by a p-value below 0.05, reflecting a reduced task completion time. Similarly, in the force-based task, the pre-knee dynamic force in Newton meters was 18.2NM, which increased to 31.26NM during knee extension post-rehabilitation. The post-student t-test showed a p-value of 0.026, signifying a significant difference. This indicated an increase in the strength of knee extensor muscles after LUNA robotic rehabilitation. Lastly, at baseline, the EMG value for ankle dorsiflexion was 5.11 (µV), which increased to 43.4 ± 0.06 µV post-rehabilitation, signifying an increase in the threshold and the patient's ability to generate more motor units during left ankle dorsiflexion. Conclusion: This study aimed to evaluate the impact of EMG and dynamic force-based rehabilitation devices on walking and strength of the affected side in chronic stroke patients without nominal data comparisons among stroke patients. Additionally, it provides insights into the inclusion of EMG-triggered neurorehabilitation robots in the daily rehabilitation of patients.Keywords: neurorehabilitation, robotic therapy, stroke, strength, paralysis
Procedia PDF Downloads 62734 Compassion Fade: Effects of Mass Perception and Intertemporal Choice on Non-Volunteering Behavior
Authors: Mariel L. Alonzo, Patricia Mae T. Chi, Juliana Patrice P. Mayormita, Sanjana A. Sorio
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Compassion fade proposes an inverse relationship between the magnitude of stimuli to elicited compassion. This phenomenon is viewed within a framework that integrates a 3-Act Compassion structure with Latané and Darley’s Unresponsive Bystander Model and Prospect Theory of Decision-making under risk. Students (N=211) from Ateneo de Davao were sampled to examine the effects of mass perception (increasing number of needy persons) and intertemporal choice (soon versus later) on volunteering behavior. Collegiate classes in their natural setting were randomly assigned to five different treatment groups and were presented with audiovisual presentations featuring an increasing number of needy persons. The students were deceived to believe that two hypothetical feeding programs for Marawi refugees, taking place in 1 month and 6 months, were in need of volunteers for its preparatory phase. Results show a statistically significant (p=0.000; p=0.013) non-linear trend consistently for both feeding programs. There was a decrease in volunteered time means as identifiable victims increased from 0-47 and an increase as it progressed towards 267 non-identifiable victims. Highest interest was expressed for the 0 needy people shown and least for 47. The 0 hours volunteered was consistently the mode and median in all treatments. There was no statistically significant temporal discounting effect.Keywords: compassion, group perception, identifiable victim, intertemporal choice, prosocial behavior, unresponsive bystander
Procedia PDF Downloads 208733 Impressions of HyFlex in an Engineering Technology Program in an Undergraduate Urban Commuter Institution
Authors: Zory Marantz
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Hybrid flexible (HyFlex) is a pedagogical methodology whereby an instructor delivers content in three modalities, i.e. live in-person (LIP), live online synchronous (LOS), and non-live online asynchronous (nLOaS). HyFlex is focused on providing the largest level of flexibility needed to achieve a cohesive environment across all modalities and incorporating four basic principles – learner’s choice, reusability, accessibility, and equivalency. Much literature has focused on the advantages of this methodology in providing students with the flexibility to choose their learning modality as best suits their schedules and learning styles. Initially geared toward graduate-level students, the concept has been applied to undergraduate studies, particularly during our national pedagogical response to the COVID19 pandemic. There is still little literature about the practicality and feasibility of HyFlex for hardware laboratory intensive engineering technology programs, particularly in dense, urban commuter institutions of higher learning. During a semester of engineering, a lab-based course was taught in the HyFlex modality, and students were asked to complete a survey about their experience. The data demonstrated that there is no single mode that is preferred by a majority of students and the usefulness of any modality is limited to how familiar the student and instructor are with the technology being applied. The technology is only as effective as our understanding and comfort with its functionality. For HyFlex to succeed in its implementation in an engineering technology environment within an urban commuter institution, faculty and students must be properly introduced to the technology being used.Keywords: education, HyFlex, technology, urban, commuter, pedagogy
Procedia PDF Downloads 95732 Influence of Stacking Sequence and Temperature on Buckling Resistance of GFRP Infill Panel
Authors: Viriyavudh Sim, SeungHyun Kim, JungKyu Choi, WooYoung Jung
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Glass Fiber Reinforced Polymer (GFRP) is a major evolution for energy dissipation when used as infill material for seismic retrofitting of steel frame, a basic PMC infill wall system consists of two GFRP laminates surrounding an infill of foam core. This paper presents numerical analysis in terms of buckling resistance of GFRP sandwich infill panels system under the influence of environment temperature and stacking sequence of laminate skin. Mode of failure under in-plane compression is studied by means of numerical analysis with ABAQUS platform. Parameters considered in this study are contact length between infill and frame, laminate stacking sequence of GFRP skin and variation of mechanical properties due to increment of temperature. The analysis is done with four cases of simple stacking sequence over a range of temperature. The result showed that both the effect of temperature and stacking sequence alter the performance of entire panel system. The rises of temperature resulted in the decrements of the panel’s strength. This is due to the polymeric nature of this material. Additionally, the contact length also displays the effect on the performance of infill panel. Furthermore, the laminate stiffness can be modified by orientation of laminate, which can increase the infill panel strength. Hence, optimal performance of the entire panel system can be obtained by comparing different cases of stacking sequence.Keywords: buckling resistance, GFRP infill panel, stacking sequence, temperature dependent
Procedia PDF Downloads 374731 A Study on the Strategy of Pocket Park in the Renewal of Old City in China
Authors: Xian Chen
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In recent years, the tendency that the decline of material and social vitality of old city in China becomes more and more serious. Nowadays, transformation and renewal of the old city have become a hot topic in urban research. The traditional mode of large-scale promotion has been criticized. Thus, exploration of new ways to update the city turns to be a necessity on the way of sustainable urban development. Pocket Park is a small city open space, its location choose is based on abandoned or idle lands on urban structure, is scattered or hidden in corner of the urban. It has a great significance on improving the old city environment. Based on the theory of ‘pocket park’, this paper summarizes the successful experience of domestic and foreign practice, and discusses the update strategies which are suitable for China's national conditions according to the characteristics and predicament of the old city in China. The main methods and results are as follows: 1)Based on the conception of ‘pocket park’, though describing the research status in domestic and foreign, summarizing the experience which is worth learning and existing problems. 2) From the analysis of ‘pocket park’ function, general design principles and types of the deep-seated difficulties in renewal the old city and the possibility of the application of ‘pocket park’,the varied implementation of ‘pocket park’ form are established, and application value in the old city renewal are summed up. 3) It can’t be denied that pocket park plays an irreplaceable role in solving the recession and renewing the vitality of the old city. Anymore, It is recommended to develop corresponding supportive development policies.Keywords: sustainable development, strategy, old city renewal, pocket park
Procedia PDF Downloads 350730 Full Characterization of Heterogeneous Antibody Samples under Denaturing and Native Conditions on a Hybrid Quadrupole-Orbitrap Mass Spectrometer
Authors: Rowan Moore, Kai Scheffler, Eugen Damoc, Jennifer Sutton, Aaron Bailey, Stephane Houel, Simon Cubbon, Jonathan Josephs
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Purpose: MS analysis of monoclonal antibodies (mAbs) at the protein and peptide levels is critical during development and production of biopharmaceuticals. The compositions of current generation therapeutic proteins are often complex due to various modifications which may affect efficacy. Intact proteins analyzed by MS are detected in higher charge states that also provide more complexity in mass spectra. Protein analysis in native or native-like conditions with zero or minimal organic solvent and neutral or weakly acidic pH decreases charge state value resulting in mAb detection at higher m/z ranges with more spatial resolution. Methods: Three commercially available mAbs were used for all experiments. Intact proteins were desalted online using size exclusion chromatography (SEC) or reversed phase chromatography coupled on-line with a mass spectrometer. For streamlined use of the LC- MS platform we used a single SEC column and alternately selected specific mobile phases to perform separations in either denaturing or native-like conditions: buffer A (20 % ACN, 0.1 % FA) with Buffer B (100 mM ammonium acetate). For peptide analysis mAbs were proteolytically digested with and without prior reduction and alkylation. The mass spectrometer used for all experiments was a commercially available Thermo Scientific™ hybrid Quadrupole-Orbitrap™ mass spectrometer, equipped with the new BioPharma option which includes a new High Mass Range (HMR) mode that allows for improved high mass transmission and mass detection up to 8000 m/z. Results: We have analyzed the profiles of three mAbs under reducing and native conditions by direct infusion with offline desalting and with on-line desalting via size exclusion and reversed phase type columns. The presence of high salt under denaturing conditions was found to influence the observed charge state envelope and impact mass accuracy after spectral deconvolution. The significantly lower charge states observed under native conditions improves the spatial resolution of protein signals and has significant benefits for the analysis of antibody mixtures, e.g. lysine variants, degradants or sequence variants. This type of analysis requires the detection of masses beyond the standard mass range ranging up to 6000 m/z requiring the extended capabilities available in the new HMR mode. We have compared each antibody sample that was analyzed individually with mixtures in various relative concentrations. For this type of analysis, we observed that apparent native structures persist and ESI is benefited by the addition of low amounts of acetonitrile and formic acid in combination with the ammonium acetate-buffered mobile phase. For analyses on the peptide level we analyzed reduced/alkylated, and non-reduced proteolytic digests of the individual antibodies separated via reversed phase chromatography aiming to retrieve as much information as possible regarding sequence coverage, disulfide bridges, post-translational modifications such as various glycans, sequence variants, and their relative quantification. All data acquired were submitted to a single software package for analysis aiming to obtain a complete picture of the molecules analyzed. Here we demonstrate the capabilities of the mass spectrometer to fully characterize homogeneous and heterogeneous therapeutic proteins on one single platform. Conclusion: Full characterization of heterogeneous intact protein mixtures by improved mass separation on a quadrupole-Orbitrap™ mass spectrometer with extended capabilities has been demonstrated.Keywords: disulfide bond analysis, intact analysis, native analysis, mass spectrometry, monoclonal antibodies, peptide mapping, post-translational modifications, sequence variants, size exclusion chromatography, therapeutic protein analysis, UHPLC
Procedia PDF Downloads 361