Search results for: free cash flow
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 7928

Search results for: free cash flow

6188 Investigation of Free Vibrations of Opened Shells from Alloy D19: Assistance of the Associated Mass System

Authors: Oleg Ye Sysoyev, Artem Yu Dobryshkin, Nyein Sitt Naing

Abstract:

Cylindrical shells are widely used in the construction of buildings and structures, as well as in the air structure. Thin-walled casings made of aluminum alloys are an effective substitute for reinforced concrete and steel structures in construction. The correspondence of theoretical calculations and the actual behavior of aluminum alloy structures is to ensure their trouble-free operation. In the laboratory of our university, "Building Constructions" conducted an experimental study to determine the effect of the system of attached masses on the natural oscillations of shallow cylindrical shells of aluminum alloys, the results of which were compared with theoretical calculations. The purpose of the experiment is to measure the free oscillations of an open, sloping cylindrical shell for various variations of the attached masses. Oscillations of an open, slender, thin-walled cylindrical shell, rectangular in plan, were measured using induction accelerometers. The theoretical calculation of the shell was carried out on the basis of the equations of motion of the theory of shallow shells, using the Bubnov-Galerkin method. A significant splitting of the flexural frequency spectrum is found, influenced not only by the systems of attached маsses but also by the values of the wave formation parameters, which depend on the relative geometric dimensions of the shell. The correspondence of analytical and experimental data is found, using the example of an open shell of alloy D19, which allows us to speak about the high quality of the study. A qualitative new analytical solution of the problem of determining the value of the oscillation frequency of the shell, carrying a system of attached masses is shown.

Keywords: open hollow shell, nonlinear oscillations, associated mass, frequency

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6187 Approximate Spring Balancing for the Arm of a Humanoid Robot to Reduce Actuator Torque

Authors: Apurva Patil, Ashay Aswale, Akshay Kulkarni, Shubham Bharadiya

Abstract:

The potential benefit of gravity compensation of linkages in mechanisms using springs to reduce actuator requirements is well recognized, but practical applications have been elusive. Although existing methods provide exact spring balance, they require additional masses or auxiliary links, or all the springs used originate from the ground, which makes the resulting device bulky and space-inefficient. This paper uses a method of static balancing of mechanisms with conservative loads such as gravity and spring loads using non-zero-free-length springs with child–parent connections and no auxiliary links. Application of this method to the developed arm of a humanoid robot is presented here. Spring balancing is particularly important in this case because the serial chain of linkages has to work against gravity.This work involves approximate spring balancing of the open-loop chain of linkages using minimization of potential energy variance. It uses the approach of flattening the potential energy distribution over the workspace and fuses it with numerical optimization. The results show the considerable reduction in actuator torque requirement with practical spring design and arrangement. Reduced actuator torque facilitates the use of lower end actuators which are generally smaller in weight and volume thereby lowering the space requirements and the total weight of the arm. This is particularly important for humanoid robots where the parent actuator has to handle the weight of the subsequent actuators as well. Actuators with lower actuation requirements are more energy efficient, thereby reduce the energy consumption of the mechanism. Lower end actuators are lower in cost and facilitate the development of low-cost devices. Although the method provides only an approximate balancing, it is versatile, flexible in choosing appropriate control variables that are relevant to the design problem and easy to implement. The true potential of this technique lies in the fact that it uses a very simple optimization to find the spring constant, free-length of the spring and the optimal attachment points subject to the optimization constraints. Also, it uses physically realizable non-zero-free-length springs directly, thereby reducing the complexity involved in simulating zero-free-length springs from non-zero-free-length springs. This method allows springs to be attached to the preceding parent link, which makes the implementation of spring balancing practical. Because auxiliary linkages can be avoided, the resultant arm of the humanoid robot is compact. The cost benefits and reduced complexity can be significant advantages in the development of this arm of the humanoid robot.

Keywords: actuator torque, child-parent connections, spring balancing, the arm of a humanoid robot

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6186 Modeling of Sand Boil near the Danube River

Authors: Edina Koch, Károly Gombás, Márton Maller

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The Little Plain is located along the Danube river, and this area is a “hotbed” of sand boil formation. This is due to the combination of a 100-250 m thick gravel layer beneath the Little Plain with a relatively thin blanket of poor soil spreading the gravel with variable thickness. Sand boils have a tradition and history in this area. It was possible to know which sand boil started and stopped working at what water level, and some of them even have names. The authors present a 2D finite element model of groundwater flow through a selected cross-section of the Danube river, which observed activation of piping phenomena during the 2013 flood event. Soil parametrization is based on a complex site investigation program conducted along the Danube River in the Little Plain.

Keywords: site characterization, groundwater flow, numerical modeling, sand boil

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6185 The Design of a Computer Simulator to Emulate Pathology Laboratories: A Model for Optimising Clinical Workflows

Authors: M. Patterson, R. Bond, K. Cowan, M. Mulvenna, C. Reid, F. McMahon, P. McGowan, H. Cormican

Abstract:

This paper outlines the design of a simulator to allow for the optimisation of clinical workflows through a pathology laboratory and to improve the laboratory’s efficiency in the processing, testing, and analysis of specimens. Often pathologists have difficulty in pinpointing and anticipating issues in the clinical workflow until tests are running late or in error. It can be difficult to pinpoint the cause and even more difficult to predict any issues which may arise. For example, they often have no indication of how many samples are going to be delivered to the laboratory that day or at a given hour. If we could model scenarios using past information and known variables, it would be possible for pathology laboratories to initiate resource preparations, e.g. the printing of specimen labels or to activate a sufficient number of technicians. This would expedite the clinical workload, clinical processes and improve the overall efficiency of the laboratory. The simulator design visualises the workflow of the laboratory, i.e. the clinical tests being ordered, the specimens arriving, current tests being performed, results being validated and reports being issued. The simulator depicts the movement of specimens through this process, as well as the number of specimens at each stage. This movement is visualised using an animated flow diagram that is updated in real time. A traffic light colour-coding system will be used to indicate the level of flow through each stage (green for normal flow, orange for slow flow, and red for critical flow). This would allow pathologists to clearly see where there are issues and bottlenecks in the process. Graphs would also be used to indicate the status of specimens at each stage of the process. For example, a graph could show the percentage of specimen tests that are on time, potentially late, running late and in error. Clicking on potentially late samples will display more detailed information about those samples, the tests that still need to be performed on them and their urgency level. This would allow any issues to be resolved quickly. In the case of potentially late samples, this could help to ensure that critically needed results are delivered on time. The simulator will be created as a single-page web application. Various web technologies will be used to create the flow diagram showing the workflow of the laboratory. JavaScript will be used to program the logic, animate the movement of samples through each of the stages and to generate the status graphs in real time. This live information will be extracted from an Oracle database. As well as being used in a real laboratory situation, the simulator could also be used for training purposes. ‘Bots’ would be used to control the flow of specimens through each step of the process. Like existing software agents technology, these bots would be configurable in order to simulate different situations, which may arise in a laboratory such as an emerging epidemic. The bots could then be turned on and off to allow trainees to complete the tasks required at that step of the process, for example validating test results.

Keywords: laboratory-process, optimization, pathology, computer simulation, workflow

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6184 Generative Design Method for Cooled Additively Manufactured Gas Turbine Parts

Authors: Thomas Wimmer, Bernhard Weigand

Abstract:

The improvement of gas turbine efficiency is one of the main drivers of research and development in the gas turbine market. This has led to elevated gas turbine inlet temperatures beyond the melting point of the utilized materials. The turbine parts need to be actively cooled in order to withstand these harsh environments. However, the usage of compressor air as coolant decreases the overall gas turbine efficiency. Thus, coolant consumption needs to be minimized in order to gain the maximum advantage from higher turbine inlet temperatures. Therefore, sophisticated cooling designs for gas turbine parts aim to minimize coolant mass flow. New design space is accessible as additive manufacturing is maturing to industrial usage for the creation of hot gas flow path parts. By making use of this technology more efficient cooling schemes can be manufacture. In order to find such cooling schemes a generative design method is being developed. It generates cooling schemes randomly which adhere to a set of rules. These assure the sanity of the design. A huge amount of different cooling schemes are generated and implemented in a simulation environment where it is validated. Criteria for the fitness of the cooling schemes are coolant mass flow, maximum temperature and temperature gradients. This way the whole design space is sampled and a Pareto optimum front can be identified. This approach is applied to a flat plate, which resembles a simplified section of a hot gas flow path part. Realistic boundary conditions are applied and thermal barrier coating is accounted for in the simulation environment. The resulting cooling schemes are presented and compared to representative conventional cooling schemes. Further development of this method can give access to cooling schemes with an even better performance having higher complexity, which makes use of the available design space.

Keywords: additive manufacturing, cooling, gas turbine, heat transfer, heat transfer design, optimization

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6183 Enhancing Aerodynamic Performance of Savonius Vertical Axis Turbine Used with Triboelectric Generator

Authors: Bhavesh Dadhich, Fenil Bamnoliya, Akshita Swaminathan

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This project aims to design a system to generate energy from flowing wind due to the motion of a vehicle on the road or from the flow of wind in compact areas to utilize the wasteful energy into a useful one. It is envisaged through a design and aerodynamic performance improvement of a Savonius vertical axis wind turbine rotor and used in an integrated system with a Triboelectric Nanogenerator (TENG) that can generate a good amount of electrical energy. Aerodynamic calculations are performed numerically using Computational Fluid Dynamics software, and TENG's performance is evaluated analytically. The Turbine's coefficient of power is validated with published results for an inlet velocity of 7 m/s with a Tip Speed Ratio of 0.75 and found to reasonably agree with that of experiment results. The baseline design is modified with a new blade arc angle and rotor position angle based on the recommended parameter ranges suggested by previous researchers. Simulations have been performed for different T.S.R. values ranging from 0.25 to 1.5 with an interval of 0.25 with two applicable free stream velocities of 5 m/s and 7m/s. Finally, the newly designed VAWT CFD performance results are used as input for the analytical performance prediction of the triboelectric nanogenerator. The results show that this approach could be feasible and useful for small power source applications.

Keywords: savonius turbine, power, overlap ratio, tip speed ratio, TENG

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6182 Clustered Regularly Interspaced Short Palindromic Repeat/cas9-Based Lateral Flow and Fluorescence Diagnostics for Rapid Pathogen Detection

Authors: Mark Osborn

Abstract:

Clustered, regularly interspaced short palindromic repeat (CRISPR/Cas) proteins can be designed to bind specified DNA and RNA sequences and hold great promise for the accurate detection of nucleic acids for diagnostics. Commercially available reagents were integrated into a CRISPR/Cas9-based lateral flow assay that can detect severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) sequences with single-base specificity. This approach requires minimal equipment and represents a simplified platform for field-based deployment. A rapid, multiplex fluorescence CRISPR/Cas9 nuclease cleavage assay capable of detecting and differentiating SARS-CoV-2, influenza A and B, and respiratory syncytial virus in a single reaction was also developed. These findings provide proof of principle for CRISPR/Cas9 point-of-care diagnosis that can detect specific SARS-CoV-2 strain(s). Further, Cas9 cleavage allows for a scalable fluorescent platform for identifying respiratory viral pathogens with overlapping symptomology. Collectively, this approach is a facile platform for diagnostics with broad application to user-defined sequence interrogation and detection.

Keywords: CRISPR/Cas9, lateral flow assay, SARS-Co-V2, single-nucleotide resolution

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6181 The Impact of Undisturbed Flow Speed on the Correlation of Aerodynamic Coefficients as a Function of the Angle of Attack for the Gyroplane Body

Authors: Zbigniew Czyz, Krzysztof Skiba, Miroslaw Wendeker

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This paper discusses the results of aerodynamic investigation of the Tajfun gyroplane body designed by a Polish company, Aviation Artur Trendak. This gyroplane has been studied as a 1:8 scale model. Scaling objects for aerodynamic investigation is an inherent procedure in any kind of designing. If scaling, the criteria of similarity need to be satisfied. The basic criteria of similarity are geometric, kinematic and dynamic. Despite the results of aerodynamic research are often reduced to aerodynamic coefficients, one should pay attention to how values of coefficients behave if certain criteria are to be satisfied. To satisfy the dynamic criterion, for example, the Reynolds number should be focused on. This is the correlation of inertial to viscous forces. With the multiplied flow speed by the specific dimension as a numerator (with a constant kinematic viscosity coefficient), flow speed in a wind tunnel research should be increased as many times as an object is decreased. The aerodynamic coefficients specified in this research depend on the real forces that act on an object, its specific dimension, medium speed and variations in its density. Rapid prototyping with a 3D printer was applied to create the research object. The research was performed with a T-1 low-speed wind tunnel (its diameter of the measurement volume is 1.5 m) and a six-element aerodynamic internal scales, WDP1, at the Institute of Aviation in Warsaw. This T-1 wind tunnel is low-speed continuous operation with open space measurement. The research covered a number of the selected speeds of undisturbed flow, i.e. V = 20, 30 and 40 m/s, corresponding to the Reynolds numbers (as referred to 1 m) Re = 1.31∙106, 1.96∙106, 2.62∙106 for the angles of attack ranging -15° ≤ α ≤ 20°. Our research resulted in basic aerodynamic characteristics and observing the impact of undisturbed flow speed on the correlation of aerodynamic coefficients as a function of the angle of attack of the gyroplane body. If the speed of undisturbed flow in the wind tunnel changes, the aerodynamic coefficients are significantly impacted. At speed from 20 m/s to 30 m/s, drag coefficient, Cx, changes by 2.4% up to 9.9%, whereas lift coefficient, Cz, changes by -25.5% up to 15.7% if the angle of attack of 0° excluded or by -25.5% up to 236.9% if the angle of attack of 0° included. Within the same speed range, the coefficient of a pitching moment, Cmy, changes by -21.1% up to 7.3% if the angles of attack -15° and -10° excluded or by -142.8% up to 618.4% if the angle of attack -15° and -10° included. These discrepancies in the coefficients of aerodynamic forces definitely need to consider while designing the aircraft. For example, if load of certain aircraft surfaces is calculated, additional correction factors definitely need to be applied. This study allows us to estimate the discrepancies in the aerodynamic forces while scaling the aircraft. This work has been financed by the Polish Ministry of Science and Higher Education.

Keywords: aerodynamics, criteria of similarity, gyroplane, research tunnel

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6180 Three-Dimensional Unsteady Natural Convection and Entropy Generation in an Inclined Cubical Trapezoidal Cavity Subjected to Uniformly Heated Bottom Wall

Authors: Farshid Fathinia

Abstract:

Numerical computation of unsteady laminar three-dimensional natural convection and entropy generation in an inclined cubical trapezoidal air-filled cavity is performed for the first time in this work. The vertical right and left sidewalls of the cavity are maintained at constant cold temperatures. The lower wall is subjected to a constant hot temperature, while the upper one is considered insulated. Computations are performed for Rayleigh numbers varied as 103 ≤ Ra ≤ 105, while the trapezoidal cavity inclination angle is varied as 0° ≤ ϕ ≤ 180°. Prandtl number is considered constant at Pr = 0.71. The second law of thermodynamics is applied to obtain thermodynamic losses inside the cavity due to both heat transfer and fluid friction irreversibilities. The variation of local and average Nusselt numbers are presented and discussed.While, streamlines, isotherms and entropy contours are presented in both two and three-dimensional pattern. The results show that when the Rayleigh number increases, the flow patterns are changed especially in three-dimensional results and the flow circulation increases. Also, the inclination angle effect on the total entropy generation becomes insignificant when the Rayleigh number is low.Moreover, when the Rayleigh number increases the average Nusselt number increases.

Keywords: transient natural convection, trapezoidal cavity, three-dimensional flow, entropy generation, second law

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6179 Resolving a Piping Vibration Problem by Installing Viscous Damper Supports

Authors: Carlos Herrera Sierralta, Husain M. Muslim, Meshal T. Alsaiari, Daniel Fischer

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Preventing piping fatigue flow induced vibration in the Oil & Gas sector demands not only the constant development of engineering design methodologies based on available software packages, but also special piping support technologies for designing safe and reliable piping systems. The vast majority of piping vibration problems in the Oil & Gas industry are provoked by the process flow characteristics which are basically intrinsically related to the fluid properties, the type of service and its different operational scenarios. In general, the corrective actions recommended for flow induced vibration in piping systems can be grouped in two major areas: those which affect the excitation mechanisms typically associated to process variables, and those which affect the response mechanism of the pipework per se, and the pipework associated steel support structure. Where possible the first option is to try to solve the flow induced problem from the excitation mechanism perspective. However, in producing facilities the approach of changing process parameters might not always be convenient as it could lead to reduction of production rates or it may require the shutdown of the system in order to perform the required piping modification. That impediment might lead to a second option, which is to modify the response of the piping system to excitation generated by the type of process flow. In principle, the action of shifting the natural frequency of the system well above the frequency inherent to the process always favours the elimination, or considerably reduces, the level of vibration experienced by the piping system. Tightening up the clearances at the supports (ideally zero gap), and adding new static supports at the system, are typical ways of increasing the natural frequency of the piping system. However, only stiffening the piping system may not be sufficient to resolve the vibration problem, and in some cases, it might not be feasible to implement it at all, as the available piping layout could create limitations on adding supports due to thermal expansion/contraction requirements. In these cases, utilization of viscous damper supports could be recommended as these devices can allow relatively large quasi-static movement of piping while providing sufficient capabilities of dissipating the vibration. Therefore, when correctly selected and installed, viscous damper supports can provide a significant effect on the response of the piping system over a wide range of frequencies. Viscous dampers cannot be used to support sustained, static loads. This paper shows over a real case example, a methodology which allows to determine the selection of the viscous damper supports via a dynamic analysis model. By implementing this methodology, it was possible to resolve the piping vibration problem throughout redesigning adequately the existing static piping supports and by adding new viscous dampers supports. This was conducted on-stream at the oil crude pipeline in question without the necessity of reducing the production of the plant. Concluding that the application of the methodology of this paper can be applied to solve similar cases in a straightforward manner.

Keywords: dynamic analysis, flow induced vibration, piping supports, turbulent flow, slug flow, viscous damper

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6178 Deployed Confidence: The Testing in Production

Authors: Shreya Asthana

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Testers know that the feature they tested on stage is working perfectly in production only after release went live. Sometimes something breaks in production and testers get to know through the end user’s bug raised. The panic mode starts when your staging test results do not reflect current production behavior. And you started doubting your testing skills when finally the user reported a bug to you. Testers can deploy their confidence on release day by testing on production. Once you start doing testing in production, you will see test result accuracy because it will be running on real time data and execution will be a little faster as compared to staging one due to elimination of bad data. Feature flagging, canary releases, and data cleanup can help to achieve this technique of testing. By this paper it will be easier to understand the steps to achieve production testing before making your feature live, and to modify IT company’s testing procedure, so testers can provide the bug free experience to the end users. This study is beneficial because too many people think that testing should be done in staging but not in production and now this is high time to pull out people from their old mindset of testing into a new testing world. At the end of the day, it all just matters if the features are working in production or not.

Keywords: bug free production, new testing mindset, testing strategy, testing approach

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6177 Wave-Assisted Flapping Foil Propulsion: Flow Physics and Scaling Laws From Fluid-Structure Interaction Simulations

Authors: Rajat Mittal, Harshal Raut, Jung Hee Seo

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Wave-assisted propulsion (WAP) systems convert wave energy into thrust using elastically mounted hydrofoils. We employ sharp-interface immersed boundary simulations to examine the effect of two key parameters on the flow physics, the fluid-structure interaction, as well as thrust performance of these systems - the stiffness of the torsional spring and the location of the rotational center. The variation in spring stiffness leads to different amplitude of pitch motion, phase difference with respect to heaving motion and thrust coefficient and we show the utility of ‘maps’ of energy exchange between the flow and the hydrofoil system, as a way to understand and predict this behavior. The Force Partitioning Method (FPM) is used to decompose the pressure forces into individual components and understand the mechanism behind increase in thrust. Next, a scaling law is presented for the thrust coefficient generated by heaving and pitching foil. The parameters within the scaling law are calculated based on direct-numerical simulations based parametric study utilized to generate the energy maps. The predictions of the proposed scaling law are then compared with those of a similar model from the literature, showing a noticeable improvement in the prediction of the thrust coefficient.

Keywords: propulsion, flapping foils, hydrodynamics, wave power

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6176 Digital Twin of Real Electrical Distribution System with Real Time Recursive Load Flow Calculation and State Estimation

Authors: Anosh Arshad Sundhu, Francesco Giordano, Giacomo Della Croce, Maurizio Arnone

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Digital Twin (DT) is a technology that generates a virtual representation of a physical system or process, enabling real-time monitoring, analysis, and simulation. DT of an Electrical Distribution System (EDS) can perform online analysis by integrating the static and real-time data in order to show the current grid status and predictions about the future status to the Distribution System Operator (DSO), producers and consumers. DT technology for EDS also offers the opportunity to DSO to test hypothetical scenarios. This paper discusses the development of a DT of an EDS by Smart Grid Controller (SGC) application, which is developed using open-source libraries and languages. The developed application can be integrated with Supervisory Control and Data Acquisition System (SCADA) of any EDS for creating the DT. The paper shows the performance of developed tools inside the application, tested on real EDS for grid observability, Smart Recursive Load Flow (SRLF) calculation and state estimation of loads in MV feeders.

Keywords: digital twin, distributed energy resources, remote terminal units, supervisory control and data acquisition system, smart recursive load flow

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6175 Dynamic Characterization of Shallow Aquifer Groundwater: A Lab-Scale Approach

Authors: Anthony Credoz, Nathalie Nief, Remy Hedacq, Salvador Jordana, Laurent Cazes

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Groundwater monitoring is classically performed in a network of piezometers in industrial sites. Groundwater flow parameters, such as direction, sense and velocity, are deduced from indirect measurements between two or more piezometers. Groundwater sampling is generally done on the whole column of water inside each borehole to provide concentration values for each piezometer location. These flow and concentration values give a global ‘static’ image of potential plume of contaminants evolution in the shallow aquifer with huge uncertainties in time and space scales and mass discharge dynamic. TOTAL R&D Subsurface Environmental team is challenging this classical approach with an innovative dynamic way of characterization of shallow aquifer groundwater. The current study aims at optimizing the tools and methodologies for (i) a direct and multilevel measurement of groundwater velocities in each piezometer and, (ii) a calculation of potential flux of dissolved contaminant in the shallow aquifer. Lab-scale experiments have been designed to test commercial and R&D tools in a controlled sandbox. Multiphysics modeling were performed and took into account Darcy equation in porous media and Navier-Stockes equation in the borehole. The first step of the current study focused on groundwater flow at porous media/piezometer interface. Huge uncertainties from direct flow rate measurements in the borehole versus Darcy flow rate in the porous media were characterized during experiments and modeling. The structure and location of the tools in the borehole also impacted the results and uncertainties of velocity measurement. In parallel, direct-push tool was tested and presented more accurate results. The second step of the study focused on mass flux of dissolved contaminant in groundwater. Several active and passive commercial and R&D tools have been tested in sandbox and reactive transport modeling has been performed to validate the experiments at the lab-scale. Some tools will be selected and deployed in field assays to better assess the mass discharge of dissolved contaminants in an industrial site. The long-term subsurface environmental strategy is targeting an in-situ, real-time, remote and cost-effective monitoring of groundwater.

Keywords: dynamic characterization, groundwater flow, lab-scale, mass flux

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6174 Hydrodynamic Characterisation of a Hydraulic Flume with Sheared Flow

Authors: Daniel Rowe, Christopher R. Vogel, Richard H. J. Willden

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The University of Oxford’s recirculating water flume is a combined wave and current test tank with a 1 m depth, 1.1 m width, and 10 m long working section, and is capable of flow speeds up to 1 ms−1 . This study documents the hydrodynamic characteristics of the facility in preparation for experimental testing of horizontal axis tidal stream turbine models. The turbine to be tested has a rotor diameter of 0.6 m and is a modified version of one of two model-scale turbines tested in previous experimental campaigns. An Acoustic Doppler Velocimeter (ADV) was used to measure the flow at high temporal resolution at various locations throughout the flume, enabling the spatial uniformity and turbulence flow parameters to be investigated. The mean velocity profiles exhibited high levels of spatial uniformity at the design speed of the flume, 0.6 ms−1 , with variations in the three-dimensional velocity components on the order of ±1% at the 95% confidence level, along with a modest streamwise acceleration through the measurement domain, a target 5 m working section of the flume. A high degree of uniformity was also apparent for the turbulence intensity, with values ranging between 1-2% across the intended swept area of the turbine rotor. The integral scales of turbulence exhibited a far higher degree of variation throughout the water column, particularly in the streamwise and vertical scales. This behaviour is believed to be due to the high signal noise content leading to decorrelation in the sampling records. To achieve more realistic levels of vertical velocity shear in the flume, a simple procedure to practically generate target vertical shear profiles in open-channel flows is described. Here, the authors arranged a series of non-uniformly spaced parallel bars placed across the width of the flume and normal to the onset flow. By adjusting the resistance grading across the height of the working section, the downstream profiles could be modified accordingly, characterised by changes in the velocity profile power law exponent, 1/n. Considering the significant temporal variation in a tidal channel, the choice of the exponent denominator, n = 6 and n = 9, effectively provides an achievable range around the much-cited value of n = 7 observed at many tidal sites. The resulting flow profiles, which we intend to use in future turbine tests, have been characterised in detail. The results indicate non-uniform vertical shear across the survey area and reveal substantial corner flows, arising from the differential shear between the target vertical and cross-stream shear profiles throughout the measurement domain. In vertically sheared flow, the rotor-equivalent turbulence intensity ranges between 3.0-3.8% throughout the measurement domain for both bar arrangements, while the streamwise integral length scale grows from a characteristic dimension on the order of the bar width, similar to the flow downstream of a turbulence-generating grid. The experimental tests are well-defined and repeatable and serve as a reference for other researchers who wish to undertake similar investigations.

Keywords: acoustic doppler Velocimeter, experimental hydrodynamics, open-channel flow, shear profiles, tidal stream turbines

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6173 Temporal and Spatio-Temporal Stability Analyses in Mixed Convection of a Viscoelastic Fluid in a Porous Medium

Authors: P. Naderi, M. N. Ouarzazi, S. C. Hirata, H. Ben Hamed, H. Beji

Abstract:

The stability of mixed convection in a Newtonian fluid medium heated from below and cooled from above, also known as the Poiseuille-Rayleigh-Bénard problem, has been extensively investigated in the past decades. To our knowledge, mixed convection in porous media has received much less attention in the published literature. The present paper extends the mixed convection problem in porous media for the case of a viscoelastic fluid flow owing to its numerous environmental and industrial applications such as the extrusion of polymer fluids, solidification of liquid crystals, suspension solutions and petroleum activities. Without a superimposed through-flow, the natural convection problem of a viscoelastic fluid in a saturated porous medium has already been treated. The effects of the viscoelastic properties of the fluid on the linear and nonlinear dynamics of the thermoconvective instabilities have also been treated in this work. Consequently, the elasticity of the fluid can lead either to a Hopf bifurcation, giving rise to oscillatory structures in the strongly elastic regime, or to a stationary bifurcation in the weakly elastic regime. The objective of this work is to examine the influence of the main horizontal flow on the linear and characteristics of these two types of instabilities. Under the Boussinesq approximation and Darcy's law extended to a viscoelastic fluid, a temporal stability approach shows that the conditions for the appearance of longitudinal rolls are identical to those found in the absence of through-flow. For the general three-dimensional (3D) perturbations, a Squire transformation allows the deduction of the complex frequencies associated with the 3D problem using those obtained by solving the two-dimensional one. The numerical resolution of the eigenvalue problem concludes that the through-flow has a destabilizing effect and selects a convective configuration organized in purely transversal rolls which oscillate in time and propagate in the direction of the main flow. In addition, by using the mathematical formalism of absolute and convective instabilities, we study the nature of unstable three-dimensional disturbances. It is shown that for a non-vanishing through-flow, general three-dimensional instabilities are convectively unstable which means that in the absence of a continuous noise source these instabilities are drifted outside the porous medium, and no long-term pattern is observed. In contrast, purely transversal rolls may exhibit a transition to absolute instability regime and therefore affect the porous medium everywhere including in the absence of a noise source. The absolute instability threshold, the frequency and the wave number associated with purely transversal rolls are determined as a function of the Péclet number and the viscoelastic parameters. Results are discussed and compared to those obtained from laboratory experiments in the case of Newtonian fluids.

Keywords: instability, mixed convection, porous media, and viscoelastic fluid

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6172 Large Amplitude Vibration of Sandwich Beam

Authors: Youssef Abdelli, Rachid Nasri

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The large amplitude free vibration analysis of three-layered symmetric sandwich beams is carried out using two different approaches. The governing nonlinear partial differential equations of motion in free natural vibration are derived using Hamilton's principle. The formulation leads to two nonlinear partial differential equations that are coupled both in axial and binding deformations. In the first approach, the method of multiple scales is applied directly to the governing equation that is a nonlinear partial differential equation. In the second approach, we discretize the governing equation by using Galerkin's procedure and then apply the shooting method to the obtained ordinary differential equations. In order to check the validity of the solutions obtained by the two approaches, they are compared with the solutions obtained by two approaches; they are compared with the solutions obtained numerically by the finite difference method.

Keywords: finite difference method, large amplitude vibration, multiple scales, nonlinear vibration

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6171 Water Desalination by Membrane Distillation with MFI Zeolite Membranes

Authors: Angelo Garofalo, Laura Donato, Maria Concetta Carnevale, Enrico Drioli, Omar Alharbi, Saad Aljlil, Alessandra Criscuoli, Catia Algieri

Abstract:

Nowadays, water scarcity may be considered one of the most important and serious questions concerning our community: in fact, there is a remarkable mismatch between water supply and water demand. Exploitation of natural fresh water resources combined with higher water demand has led to an increased requirement for alternative water resources. In this context, desalination provides such an alternative source, offering water otherwise not accessible for irrigational, industrial and municipal use. Considering the various drawbacks of the polymeric membranes, zeolite membranes represent a potential device for water desalination owing to their high thermal and chemical stability. In this area wide attention was focused on the MFI (silicalite, ZSM-5) membranes, having a pore size lower (about 5.5 Å) than the major kinetic diameters of hydrated ions. In the present work, a scale-up for the preparation of supported silicalite membranes was performed. Therefore, tubular membranes 30 cm long were synthesized by using the secondary growth method coupled with the cross flow seeding procedure. The secondary growth presents two steps: seeding and growth of zeolite crystals on the support. This process, decoupling zeolite nucleation from crystals growth, permits to control the conditions of each step separately. The seeding procedure consists of a cross-flow filtration through a porous support coupled with the support rotation and tilting. The combination of these three different aspects allows a homogeneous and uniform coverage of the support with the zeolite seeds. After characterization by scanning electron microscope (SEM), X-ray diffractometry (XRD) and Energy-dispersive X-ray (EDX) analysis, the prepared membranes were tested by means of single gas permeation and then by Vacuum Membrane Distillation (VMD) using both deionized water and NaCl solutions. The experimental results evidenced the possibility to perform the scale up for the preparation of almost defect free silicalite membranes. VMD tests indicated the possibility to prepare membranes that exhibit interesting performance in terms of fluxes and salt rejections for concentrations from 0.2 M to 0.9 M. Furthermore, it was possible to restore the original performance of the membrane after an identified cleaning procedure. Acknowledgements: The authors gratefully acknowledge the support of the King Abdulaziz City for Science and Technology (KACST) for funding the research Project 895/33 entitled ‘Preparation and Characterization of Zeolite Membranes for Water Treatment’.

Keywords: desalination, MFI membranes, secondary growth, vacuum membrane distillation

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6170 Cell Surface Display of Xylanase on Escherichia coli by TibA Autotransporter

Authors: Yeng Min Yi, Rosli Md Illias, Salehhuddin Hamdan

Abstract:

Industrial biocatalysis is mainly based on the use of cell free or intracellular enzyme systems. However, the expensive cost and relatively lower operational stability of free enzymes limit practical use in industries. Cell surface display system can be used as a cost-efficient alternative to overcome the laborious purification and substrate transport limitation. In this research, TibA autotransporter from E. coli was used to display Aspergillus fumigatus xylanase (xyn). The amplified xyn was fused in between N-terminal signal peptide and C-terminal β-barrel of TibA. The cloned was transformed and expressed in E. coli BL21 (DE3). Outer membrane localization of TibA-xyn fusion protein was confirmed by SDS PAGE and western blot with expected size of 62.5 kDa. Functional display of xyn was examined by activity assay. Cell surface displayed xyn exhibited the highest activity at 37 °c, 0.3 mM IPTG. As a summary, TibA displaying system has the potential for further industrial applications. Moreover, this is the first report of the display of xylanase using TibA on the surface of E. coli.

Keywords: biocatalysis, cell surface display, Escherichia coli, TibA autotransporter

Procedia PDF Downloads 261
6169 Comparison of Safety and Efficacy between Thulium Fibre Laser and Holmium YAG Laser for Retrograde Intrarenal Surgery

Authors: Sujeet Poudyal

Abstract:

Introduction: After Holmium:yttrium-aluminum-garnet (Ho: YAG) laser has revolutionized the management of urolithiasis, the introduction of Thulium fibre laser (TFL) has already challenged Ho:YAG laser due to its multiple commendable properties. Nevertheless, there are only few studies comparing TFL and holmium laser in Retrograde Intrarenal Surgery(RIRS). Therefore, this study was carried out to compare the efficacy and safety of thulium fiber laser (TFL) and holmium laser in RIRS. Methods: This prospective comparative study, which included all patients undergoing laser lithotripsy (RIRS) for proximal ureteric calculus and nephrolithiasis from March 2022 to March 2023, consisted of 63 patients in Ho:YAG laser group and 65 patients in TFL group. Stone free rate, operative time, laser utilization time, energy used, and complications were analysed between the two groups. Results: Mean stone size was comparable in TFL (14.23±4.1 mm) and Ho:YAG (13.88±3.28 mm) group, p-0.48. Similarly, mean stone density in TFL (1269±262 HU) was comparable to Ho:YAG (1189±212 HU), p-0.48. There was significant difference in lasing time between TFL (12.69±7.41 mins) and Ho:YAG (20.44±14 mins), p-0.012). TFL group had operative time of 43.47± 16.8 mins which was shorter than Ho:YAG group (58±26.3 mins),p-0.005. Both TFL and Ho:YAG groups had comparable total energy used(11.4±6.2 vs 12±8 respectively, p-0.758). Stone free rate was 87%for TFL, whereas it was 79.5% for Ho:YAG, p-0.25). Two cases of sepsis and one ureteric stricture were encountered in TFL, whereas three cases suffered from sepsis apart from one ureteric stricture in Ho:YAG group, p-0.62). Conclusion: Thulium Fibre Laser has similar efficacy as Holmium: YAG Laser in terms of safety and stone free rate. However, due to better stone ablation rate in TFL, it can become the game changer in management of urolithiasis in the coming days.

Keywords: retrograde intrarenal surgery, thulium fibre laser, holmium:yttrium-aluminum-garnet (ho:yag) laser, nephrolithiasis

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6168 Flow Field Analysis of Different Intake Bump (Compression Surface) Configurations on a Supersonic Aircraft

Authors: Mudassir Ghafoor, Irsalan Arif, Shuaib Salamat

Abstract:

This paper presents modeling and analysis of different intake bump (compression surface) configurations and comparison with an existing supersonic aircraft having bump intake configuration. Many successful aircraft models have shown that Diverter less Supersonic Inlet (DSI) as compared to conventional intake can reduce weight, complexity and also maintenance cost. The research is divided into two parts. In the first part, four different intake bumps are modeled for comparative analysis keeping in view the consistency of outer perimeter dimensions of fighter aircraft and various characteristics such as flow behavior, boundary layer diversion and pressure recovery are analyzed. In the second part, modeled bumps are integrated with intake duct for performance analysis and comparison with existing supersonic aircraft data is carried out. The bumps are named as uniform large (Config 1), uniform small (Config 2), uniform sharp (Config 3), non-uniform (Config 4) based on their geometric features. Analysis is carried out at different Mach Numbers to analyze flow behavior in subsonic and supersonic regime. Flow behavior, boundary layer diversion and Pressure recovery are examined for each bump characteristics, and comparative study is carried out. The analysis reveals that at subsonic speed, Config 1 and Config 2 give similar pressure recoveries as diverterless supersonic intake, but difference in pressure recoveries becomes significant at supersonic speed. It was concluded from research that Config 1 gives better results as compared to Config 3. Also, higher amplitude (Config 1) is preferred over lower (Config 2 and 4). It was observed that maximum height of bump is preferred to be placed near cowl lip of intake duct.

Keywords: bump intake, boundary layer, computational fluid dynamics, diverter-less supersonic inlet

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6167 Experimental Characterization of Flowable Cement Pastes Made with Marble Waste

Authors: F. Messaoudi, O. Haddad, R. Bouras, S. Kaci

Abstract:

The development of self-compacting concrete (SCC) marks a huge step towards improved efficiency and working conditions on construction sites and in the precast industry. SCC flows easily into more complex shapes and through reinforcement bars, reduces the manpower required for the placement; no vibration is required to ensure correct compaction of concrete. This concrete contains a high volume of binder which is controlled by their rheological behavior. The paste consists of binders (Portland cement with or without supplementary cementitious materials), water, chemical admixtures and fillers. In this study, two series of tests were performed on self-compacting cement pastes made with marble waste additions as the mineral addition. The first series of this investigation was to determine the flow time of paste using Marsh cone, the second series was to determine the rheological parameters of the same paste namely yield stress and plastic viscosity using the rheometer Haake RheoStress 1. The results of this investigation allowed us to study the evolution of the yield stress, viscosity and the flow time Marsh cone paste as a function of the composition of the paste. A correlation between the results obtained on the flow test Marsh cone and those of the plastic viscosity on the mottled different cement pastes is proposed.

Keywords: adjuvant, rheological parameter, self-compacting cement pastes, waste marble

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6166 Reduction of Specific Energy Consumption in Microfiltration of Bacillus velezensis Broth by Air Sparging and Turbulence Promoter

Authors: Jovana Grahovac, Ivana Pajcin, Natasa Lukic, Jelena Dodic, Aleksandar Jokic

Abstract:

To obtain purified biomass to be used in the plant pathogen biocontrol or as soil biofertilizer, it is necessary to eliminate residual broth components at the end of the fermentation process. The main drawback of membrane separation techniques is permeate flux decline due to the membrane fouling. Fouling mitigation measures increase the pressure drop along membrane channel due to the increased resistance to flow of the feed suspension, thus increasing the hydraulic power drop. At the same time, these measures lead to an increase in the permeate flux due to the reduced resistance of the filtration cake on the membrane surface. Because of these opposing effects, the energy efficiency of fouling mitigation measures is limited, and the justification of its application is provided by information on a reducing specific energy consumption compared to a case without any measures employed. In this study, the influence of static mixer (Kenics) and air-sparging (two-phase flow) on reduction of specific energy consumption (ER) was investigated. Cultivation Bacillus velezensis was carried out in the 3-L bioreactor (Biostat® Aplus) containing 2 L working volume with two parallel Rushton turbines and without internal baffles. Cultivation was carried out at 28 °C on at 150 rpm with an aeration rate of 0.75 vvm during 96 h. The experiments were carried out in a conventional cross-flow microfiltration unit. During experiments, permeate and retentate were recycled back to the broth vessel to simulate continuous process. The single channel ceramic membrane (TAMI Deutschland) used had a nominal pore size 200 nm with the length of 250 mm and an inner/external diameter of 6/10 mm. The useful membrane channel surface was 4.33×10⁻³ m². Air sparging was brought by the pressurized air connected by a three-way valve to the feed tube by a simple T-connector without diffusor. The different approaches to flux improvement are compared in terms of energy consumption. Reduction of specific energy consumption compared to microfiltration without fouling mitigation is around 49% and 63%, for use of two-phase flow and a static mixer, respectively. In the case of a combination of these two fouling mitigation methods, ER is 60%, i.e., slightly lower compared to the use of turbulence promoter alone. The reason for this result can be found in the fact that flux increase is more affected by the presence of a Kenics static mixer while sparging results in an increase of energy used during microfiltration. By comparing combined method with turbulence promoter flux enhancement method ER is negative (-7%) which can be explained by increased power consumption for air flow with moderate contribution to the flux increase. Another confirmation for this fact can be found by comparing energy consumption values for combined method with energy consumption in the case of two-phase flow. In this instance energy reduction (ER) is 22% that demonstrates that turbulence promoter is more efficient compared to two phase flow. Antimicrobial activity of Bacillus velezensis biomass against phytopathogenic isolates Xanthomonas campestris was preserved under different fouling reduction methods.

Keywords: Bacillus velezensis, microfiltration, static mixer, two-phase flow

Procedia PDF Downloads 106
6165 Ta-DAH: Task Driven Automated Hardware Design of Free-Flying Space Robots

Authors: Lucy Jackson, Celyn Walters, Steve Eckersley, Mini Rai, Simon Hadfield

Abstract:

Space robots will play an integral part in exploring the universe and beyond. A correctly designed space robot will facilitate OOA, satellite servicing and ADR. However, problems arise when trying to design such a system as it is a highly complex multidimensional problem into which there is little research. Current design techniques are slow and specific to terrestrial manipulators. This paper presents a solution to the slow speed of robotic hardware design, and generalizes the technique to free-flying space robots. It presents Ta-DAH Design, an automated design approach that utilises a multi-objective cost function in an iterative and automated pipeline. The design approach leverages prior knowledge and facilitates the faster output of optimal designs. The result is a system that can optimise the size of the base spacecraft, manipulator and some key subsystems for any given task. Presented in this work is the methodology behind Ta-DAH Design and a number optimal space robot designs.

Keywords: space robots, automated design, on-orbit operations, hardware design

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6164 The Effect of Improvement Programs in the Mean Time to Repair and in the Mean Time between Failures on Overall Lead Time: A Simulation Using the System Dynamics-Factory Physics Model

Authors: Marcel Heimar Ribeiro Utiyama, Fernanda Caveiro Correia, Dario Henrique Alliprandini

Abstract:

The importance of the correct allocation of improvement programs is of growing interest in recent years. Due to their limited resources, companies must ensure that their financial resources are directed to the correct workstations in order to be the most effective and survive facing the strong competition. However, to our best knowledge, the literature about allocation of improvement programs does not analyze in depth this problem when the flow shop process has two capacity constrained resources. This is a research gap which is deeply studied in this work. The purpose of this work is to identify the best strategy to allocate improvement programs in a flow shop with two capacity constrained resources. Data were collected from a flow shop process with seven workstations in an industrial control and automation company, which process 13.690 units on average per month. The data were used to conduct a simulation with the System Dynamics-Factory Physics model. The main variables considered, due to their importance on lead time reduction, were the mean time between failures and the mean time to repair. The lead time reduction was the output measure of the simulations. Ten different strategies were created: (i) focused time to repair improvement, (ii) focused time between failures improvement, (iii) distributed time to repair improvement, (iv) distributed time between failures improvement, (v) focused time to repair and time between failures improvement, (vi) distributed time to repair and between failures improvement, (vii) hybrid time to repair improvement, (viii) hybrid time between failures improvements, (ix) time to repair improvement strategy towards the two capacity constrained resources, (x) time between failures improvement strategy towards the two capacity constrained resources. The ten strategies tested are variations of the three main strategies for improvement programs named focused, distributed and hybrid. Several comparisons among the effect of the ten strategies in lead time reduction were performed. The results indicated that for the flow shop analyzed, the focused strategies delivered the best results. When it is not possible to perform a large investment on the capacity constrained resources, companies should use hybrid approaches. An important contribution to the academy is the hybrid approach, which proposes a new way to direct the efforts of improvements. In addition, the study in a flow shop with two strong capacity constrained resources (more than 95% of utilization) is an important contribution to the literature. Another important contribution is the problem of allocation with two CCRs and the possibility of having floating capacity constrained resources. The results provided the best improvement strategies considering the different strategies of allocation of improvement programs and different positions of the capacity constrained resources. Finally, it is possible to state that both strategies, hybrid time to repair improvement and hybrid time between failures improvement, delivered best results compared to the respective distributed strategies. The main limitations of this study are mainly regarding the flow shop analyzed. Future work can further investigate different flow shop configurations like a varying number of workstations, different number of products or even different positions of the two capacity constrained resources.

Keywords: allocation of improvement programs, capacity constrained resource, hybrid strategy, lead time, mean time to repair, mean time between failures

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6163 Intelligent Ambulance with Advance Features of Traffic Management and Telecommunication

Authors: Mamatha M. N.

Abstract:

Traffic problems, congested traffic, and flow management were recognized as major problems mostly in all the areas, which have caused a problem for the ambulance which carries the emergency patient. The proposed paper aims in the development of ambulance which reaches the nearby hospital faster even in heavy traffic scenario. This process is activated by implementing hardware in an ambulance as well as in traffic post thus allowing a smooth flow to the ambulance to reach the hospital in time. 1) The design of the vehicle to have a communication between ambulance and traffic post. 2)Electronic Health Record with Data-acquisition system 3)Telemetry of acquired biological parameters to the nearest hospital. Thus interfacing all these three different modules and integrating them on the ambulance could reach the hospital earlier than the present ambulance. The system is accurate and efficient of 99.8%.

Keywords: bio-telemetry, data acquisition, patient database, automatic traffic control

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6162 A Low Power and High-Speed Conditional-Precharge Sense Amplifier Based Flip-Flop Using Single Ended Latch

Authors: Guo-Ming Sung, Ramavath Naga Raju Naik

Abstract:

This paper presents a low power, high speed, sense-amplifier based flip-flop (SAFF). The flip-flop’s power con-sumption and delay are greatly reduced by employing a new conditionally precharge sense-amplifier stage and a single-ended latch stage. Glitch-free and contention-free latch operation is achieved by using a conditional cut-off strategy. The design uses fewer transistors, has a lower clock load, and has a simple structure, all of which contribute to a near-zero setup time. When compared to previous flip-flop structures proposed for similar input/output conditions, this design’s performance and overall PDP have improved. The post layout simulation of the circuit uses 2.91µW of power and has a delay of 65.82 ps. Overall, the power-delay product has seen some enhancements. Cadence Virtuoso Designing tool with CMOS 90nm technology are used for all designs.

Keywords: high-speed, low-power, flip-flop, sense-amplifier

Procedia PDF Downloads 142
6161 Learning through Gaming with Mobile Devices

Authors: Luis Rodrigo Valencia Pérez, Juan Manuel Peña Aguilar, Adelina Morita Alexander, Alberto Lamadrid Alvarez, Héctor Fernando Valencia Pérez

Abstract:

Financial education is among the areas of opportunity in the Spanish-speaking from an early age to high school, through mobile devices such as cell phones and tablets using ludic and fun applications like interactive games, children can learn money management and investment through time, thereby fostering the habit of saving and/or sound management of cash and family business resources, having interaction with an uncontrolled environment such as the involvement of other players in the external decisions of the environment in which the game is play. The application proposed in Phase 1 (design and development) was designed in multi-user environments, under methodologies of hybrid programming for any platform on the market and designed under CMMI standards that allow for quality production over time, following up on these improvements counting with continuous user feedback and usage statistics.

Keywords: mobile educational games, ludic games, children, multiuser, design and software development

Procedia PDF Downloads 360
6160 An Investigation of Item Bias in Free Boarding and Scholarship Examination in Turkey

Authors: Yeşim Özer Özkan, Fatma Büşra Fincan

Abstract:

Biased sample is a regression of an observation, design process and all of the specifications lead to tendency of a side or the situation of leaving from the objectivity. It is expected that, test items are answered by the students who come from different social groups and the same ability not to be different from each other. The importance of the expectation increases especially during student selection and placement examinations. For example, all of the test items should not be beneficial for just a male or female group. The aim of the research is an investigation of item bias whether or not the exam included in 2014 free boarding and scholarship examination in terms of gender variable. Data which belong to 5th, 6th, and 7th grade the secondary education students were obtained by the General Directorate of Measurement, Evaluation and Examination Services in Turkey. 20% students were selected randomly within 192090 students. Based on 38418 students’ exam paper were examined for determination item bias. Winsteps 3.8.1 package program was used to determine bias in analysis of data, according to Rasch Model in respect to gender variable. Mathematics items tests were examined in terms of gender bias. Firstly, confirmatory factor analysis was applied twenty-five math questions. After that, NFI, TLI, CFI, IFI, RFI, GFI, RMSEA, and SRMR were examined in order to be validity and values of goodness of fit. Modification index values of confirmatory factor analysis were examined and then some of the items were omitted because these items gave an error in terms of model conformity and conceptual. The analysis shows that in 2014 free boarding and scholarship examination exam does not include bias. This is an indication of the gender of the examination to be made in favor of or against different groups of students.

Keywords: gender, item bias, placement test, Rasch model

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6159 Cell-free Bioconversion of n-Octane to n-Octanol via a Heterogeneous and Bio-Catalytic Approach

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

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

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

Procedia PDF Downloads 208