Search results for: tanner EDA tool write access time and retention time

Authors: R. Uporn, P. Luangpaiboon

Abstract:

This paper presents a comparative study of statistical methods for the multi-response surface optimization of a cryogenic freezing process. Taguchi design and analysis and steepest ascent methods based on the desirability function were conducted to ascertain the influential factors of a cryogenic freezing process and their optimal levels. The more preferable levels of the set point, exhaust fan speed, retention time and flow direction are set at -90oC, 20 Hz, 18 minutes and Counter Current, respectively. The overall desirability level is 0.7044.

Keywords: Cryogenic Freezing Process, Taguchi Design and Analysis, Response Surface Method, Steepest Ascent Method and Desirability Function Approach.

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Authors: Kevser Simsek, Rahime Gunay

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In this paper, we focused primarily on Istanbul data that is gathered by using intelligent transportation systems (ITS), and considered the developments in traffic information delivery and future applications that are being planned for implementation. Since traffic congestion is increasing and travel times are becoming less consistent and less predictable, traffic information delivery has become a critical issue. Considering the fuel consumption and wasted time in traffic, advanced traffic information systems are becoming increasingly valuable which enables travelers to plan their trips more accurately and easily.

Keywords: Data Fusion, Istanbul, ITS, Real Time Information, Traffic Information, Travel Time, Urban Mobility

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Authors: Jan Busch, Maurice Schmidt, Peter Nyhuis

Abstract:

The change of conditions for production companies in high-wage countries is characterized by the globalization of competition and the transition of a supplier´s to a buyer´s market. The companies need to face the challenges of reacting flexibly to these changes. Due to the significant and increasing degree of automation, assembly has become the most expensive production process. Regarding the reduction of production cost, assembly consequently offers a considerable rationalizing potential. Therefore, an aerodynamic feeding system has been developed at the Institute of Production Systems and Logistics (IFA), Leibniz Universitaet Hannover. This system has been enabled to adjust itself by using a genetic algorithm. The longer this genetic algorithm is executed the better is the feeding quality. In this paper, the relation between the system´s setting time and the feeding quality is observed and a function which enables the user to achieve the minimum of the total feeding time is presented.

Keywords: Aerodynamic feeding system, batch size, optimisation, setting time.

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Authors: Sahar S. Tabrizi, Dogan Ibrahim

Abstract:

Cloud Computing is a convenient model for on-demand networks that uses shared pools of virtual configurable computing resources, such as servers, networks, storage devices, applications, etc. The cloud serves as an environment for companies and organizations to use infrastructure resources without making any purchases and they can access such resources wherever and whenever they need. Cloud computing is useful to overcome a number of problems in various Information Technology (IT) domains such as Geographical Information Systems (GIS), Scientific Research, e-Governance Systems, Decision Support Systems, ERP, Web Application Development, Mobile Technology, etc. Companies can use Cloud Computing services to store large amounts of data that can be accessed from anywhere on Earth and also at any time. Such services are rented by the client companies where the actual rent depends upon the amount of data stored on the cloud and also the amount of processing power used in a given time period. The resources offered by the cloud service companies are flexible in the sense that the user companies can increase or decrease their storage requirements or the processing power requirements at any time, thus minimizing the overall rental cost of the service they receive. In addition, the Cloud Computing service providers offer fast processors and applications software that can be shared by their clients. This is especially important for small companies with limited budgets which cannot afford to purchase their own expensive hardware and software. This paper is an overview of the Cloud Computing, giving its types, principles, advantages, and disadvantages. In addition, the paper gives some example engineering applications of Cloud Computing and makes suggestions for possible future applications in the field of engineering.

Keywords: Cloud computing, cloud services, IaaS, PaaS, SaaS, IoT.

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Authors: YaoAn Lu, QingZhen Bi, BaoRui Du, ShuLin Chen, LiMin Zhu, Kai Huang

Abstract:

Taking the design tolerance into account, this paper presents a novel efficient approach to generate iso-scallop tool path for five-axis strip machining with a barrel cutter. The cutter location is first determined on the scallop surface instead of the design surface, and then the cutter is adjusted to locate the optimal tool position based on the differential rotation of the tool axis and satisfies the design tolerance simultaneously. The machining strip width and error are calculated with the aid of the grazing curve of the cutter. Based on the proposed tool positioning algorithm, the tool paths are generated by keeping the scallop height formed by adjacent tool paths constant. An example is conducted to confirm the validity of the proposed method.

Keywords: Strip machining, barrel cutter, iso-scallop tool path, sculptured surfaces, differential motion.

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Authors: Keunhong Chae, Seokho Yoon

Abstract:

This paper proposes a cooperative Alamouti space time transmission scheme with low relay complexity for the cooperative communication systems. In the proposed scheme, the source node combines the data symbols to construct the Alamouti-coded form at the destination node, while the conventional scheme performs the corresponding operations at the relay nodes. In simulation results, it is shown that the proposed scheme achieves the second order cooperative diversity while maintaining the same bit error rate (BER) performance as that of the conventional scheme.

Keywords: Space-time transmission, cooperative communication system, MIMO.

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Authors: Amin Zabihinejad, Philippe Viarouge

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In this paper, a direct power control (DPC) strategies have been investigated in order to control a high power AC/DC converter with time variable load. This converter is composed of a three level three phase neutral point clamped (NPC) converter as rectifier and an H-bridge four quadrant current control converter. In the high power application, controller not only must adjust the desire outputs but also decrease the level of distortions which are injected to the network from the converter. Regarding to this reason and nonlinearity of the power electronic converter, the conventional controllers cannot achieve appropriate responses. In this research, the precise mathematical analysis has been employed to design the appropriate controller in order to control the time variable load. A DPC controller has been proposed and simulated using Matlab/ Simulink. In order to verify the simulation result, a real time simulator- OPAL-RT- has been employed. In this paper, the dynamic response and stability of the high power NPC with variable load has been investigated and compared with conventional types using a real time simulator. The results proved that the DPC controller is more stable and has more precise outputs in comparison with conventional controller.

Keywords: Direct Power Control, Three Level Rectifier, Real Time Simulator, High Power Application.

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Authors: Siliang Wang, Minghui Wang, Jun Hu

Abstract:

A novel path planning approach is presented to solve optimal path in stochastic, time-varying networks under priori traffic information. Most existing studies make use of dynamic programming to find optimal path. However, those methods are proved to be unable to obtain global optimal value, moreover, how to design efficient algorithms is also another challenge. This paper employs a decision theoretic framework for defining optimal path: for a given source S and destination D in urban transit network, we seek an S - D path of lowest expected travel time where its link travel times are discrete random variables. To solve deficiency caused by the methods of dynamic programming, such as curse of dimensionality and violation of optimal principle, an integer programming model is built to realize assignment of discrete travel time variables to arcs. Simultaneously, pruning techniques are also applied to reduce computation complexity in the algorithm. The final experiments show the feasibility of the novel approach.

Keywords: pruning method, stochastic, time-varying networks, optimal path planning.

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Authors: P. Bárta

Abstract:

Increasing demand on the performance of Subsea Production Systems (SPS) suggests a need for more detailed investigation of fluid behavior taking place in subsea equipment. Complete CFD cool down analyses of subsea equipment are very time demanding. The objective of this paper is to investigate a Locked CFD approach, which enables significant reduction of the computational time and at the same time maintains sufficient accuracy during thermal cool down simulations. The result comparison of a dead leg simulation using the Full CFD and the three LCFD-methods confirms the validity of the locked flow field assumption for the selected case. For the tested case the LCFD simulation speed up by factor of 200 results in the absolute thermal error of 0.5 °C (3% relative error), speed up by factor of 10 keeps the LCFD results within 0.1 °C (0.5 % relative error) comparing to the Full CFD.

Keywords: CFD, Locked Flow Field, Speed up of CFD simulation time, Subsea

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Authors: Roengruen P., Tipsuwanporn V., Puawade P., Numsomran A.

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Smith Predictor control is theoretically a good solution to the problem of controlling the time delay systems. However, it seldom gets use because it is almost impossible to find out a precise mathematical model of the practical system and very sensitive to uncertain system with variable time-delay. In this paper is concerned with a design method of smith predictor for temperature control system by Coefficient Diagram Method (CDM). The simulation results show that the control system with smith predictor design by CDM is stable and robust whilst giving the desired time domain system performance.

Keywords: CDM, Smith Predictor, temperature process

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Authors: Hafiz Usama Imad, Muhammad Akram Akhund, Tauha Hussain Ali, Ali Raza Khoso, Fida Hussain Siddiqui

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Construction industry plays a significant role in fulfilling the major requirements of the human being. It is one of the major constituents of every developed country. Although the construction industry of both the developing and developed countries encompasses a major part of the economy, and millions of rupees are utilized every year on various kinds of construction projects. But, this industry is facing numerous hurdles in terms of its budget and timely completion. Construction projects generally consist of several phases like planning, designing, execution, and finishing. This research study aims to determine the significant factors of time overrun in pre-construction planning (PCP) phase of construction projects in Pakistan. Questionnaires were distributed by various means and responses of respondents were compiled and collected data were then analyzed through a statistical technique using SPSS version 24. Major causes of time overrun in pre-construction planning phase; which is an extremely important phase of construction projects, were revealed. The research conclusion will provide a pathway for stakeholders to pay attention to the mentioned causes to overcome the major issue of time overrun.

Keywords: Construction industry, Pakistan, pre-construction planning phase, time overrun.

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Authors: Alonggot Limcharoen, Jintana Wannarat, Vorawat Panich

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This study aims to investigate the balancing of the number of operators (Line Balancing technique) in the production line of hard disk drive components in order to increase efficiency. At present, the trend of using hard disk drives has continuously declined leading to limits in a company’s revenue potential. It is important to improve and develop the production process to create market share and to have the ability to compete with competitors with a higher value and quality. Therefore, an effective tool is needed to support such matters. In this research, the Arena program was applied to analyze the results both before and after the improvement. Finally, the precedent was used before proceeding with the real process. There were 14 work stations with 35 operators altogether in the RA production process where this study was conducted. In the actual process, the average production time was 84.03 seconds per product piece (by timing 30 times in each work station) along with a rating assessment by implementing the Westinghouse principles. This process showed that the rating was 123% underlying an assumption of 5% allowance time. Consequently, the standard time was 108.53 seconds per piece. The Takt time was calculated from customer needs divided by working duration in one day; 3.66 seconds per piece. Of these, the proper number of operators was 30 people. That meant five operators should be eliminated in order to increase the production process. After that, a production model was created from the actual process by using the Arena program to confirm model reliability; the outputs from imitation were compared with the original (actual process) and this comparison indicated that the same output meaning was reliable. Then, worker numbers and their job responsibilities were remodeled into the Arena program. Lastly, the efficiency of production process enhanced from 70.82% to 82.63% according to the target.

Keywords: Hard disk drive, line balancing, simulation, Arena program.

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Authors: K. Santacruz-Gomez, E. Silva-Campa, S. Álvarez-García, V. Mata-Haro, D. Soto-Puebla, M. Pedroza-Montero

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Blood gamma irradiation is the only available method to prevent transfusion associated graft versus host disease (TAGVHD). However, when blood is irradiated, determine blood shelf time is crucial. Non irradiated blood have a self-time from 21 to 35 days when is preserved with anticoagulated solution and stored at 4°C. During their storage, red blood cells (RBC) undergo a series of biochemical, biomechanical and molecular changes involving what is known as storage lesion (SL). SL include loss of structural integrity of RBC, decrease of 2,3-diphosphatidylglyceric acid levels, and increase of both ion potassium concentration and hemoglobin (Hb). On the other hand, Atomic force Microscopy (AFM) represents a versatile tool for a nano-scale high resolution topographic analysis in biological systems. In order to evaluate SL in irradiated and nonirradiated blood, RBC topography and morphometric parameters were obtained from an AFM XE-BIO system. Cell viability was followed using flow cytometry. Our results showed that early markers as nanoscale roughness, allow us to evaluate blood quality since other perspective.

Keywords: AFM, Blood γ-irradiation, roughness, Storage lesion.

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Authors: Shi-hong Lu, Chao-xun Liu, Yi-feng Zhu

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Ultrasonic shot peening (USP) on AALY12 sheet was studied. Several parameters (arc heights, surface roughness, surface topography and micro hardness) with different USP process parameters were measured. The research proposes that radius of curvature of shot peened sheet increases with time and electric current decreasing, while increases with pin diameter increasing, and radius of curvature reaches a saturation level after a specific processing time and electric current. An empirical model of the relationship between radius of curvature and pin diameter, electric current, time was also obtained. The research shows that the increment of surface and vertical micro hardness of material is more obvious with longer time and higher value of electric current, which can be up to 20% and 28% respectively.

Keywords: USP forming, surface properties, radius of curvature, residual stress.

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Authors: A. Chandrashekar, H. N. Reddappa, B. S. Ajaykumar

Abstract:

A Friction stir welding tool is a critical component to the success of the process. The tool typically consists of a rotating round shoulder and a threaded cylindrical pin that heats the work piece, mostly by friction, and moves the softened alloy around it to form the joint. In this research work, an attempt has been made to investigate the relationship between FSW variables mainly tool profile, rotating speed, welding speed and the mechanical properties (tensile strength, yield strength, percentage elongation, and micro hardness) of friction stir welded aluminum alloy 5083 joints. From the experimental details, it can be assessed that the joint produced by using Triflute profile tool has contribute superior mechanical and structural properties as compared to Tapered unthreaded & Threaded tool for 1000rpm.

Keywords: Friction stir welding, Tool profile, Rotating speed, Strength, Speed ratio.

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Authors: Amira Mabrouk, Chokri Abdennadher

Abstract:

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, value of time, contingent valuation, time value, city toll, transport.

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Authors: M. Esmailian, M. Shakouri, A. Mottahedi, S. G. Shabestari

Abstract:

Heat treatable aluminum alloys such as 7075 and 7055, because of high strength and low density, are used widely in aircraft industry. For best mechanical properties, T6 heat treatment has recommended for this regards, but this temper treatment is sensitive to corrosion induced and Stress Corrosion Cracking (SCC) damage. For improving this property, the over-aging treatment (T7) applies to this alloy, but it decreases the mechanical properties up to 30 percent. Hence, to increase the mechanical properties, without any remarkable decrease in SCC resistant, Retrogression and Re-Aging (RRA) heat treatment is used. This treatment performs in a relatively short time. In this paper, the RRA heat treatment was applied to 7055 aluminum alloy and then effect of RRA time on the mechanical properties of 7055 has been investigated. The results show that the 40-minute time is suitable time for retrogression of 7055 aluminum alloy and ultimate strength increases up to 625MPa.

Keywords: 7055 Aluminum alloy, Mechanical properties, SCC resistance, Heat Treatment.

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Authors: Chidentree Treesatayapun

Abstract:

A direct adaptive controller for a class of unknown nonlinear discrete-time systems is presented in this article. The proposed controller is constructed by fuzzy rules emulated network (FREN). With its simple structure, the human knowledge about the plant is transferred to be if-then rules for setting the network. These adjustable parameters inside FREN are tuned by the learning mechanism with time varying step size or learning rate. The variation of learning rate is introduced by main theorem to improve the system performance and stabilization. Furthermore, the boundary of adjustable parameters is guaranteed through the on-line learning and membership functions properties. The validation of the theoretical findings is represented by some illustrated examples.

Keywords: Neuro-Fuzzy, learning algorithm, nonlinear discrete time.

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Authors: Zhengnan Shi

Abstract:

In the self-stabilizing algorithmic paradigm, each node has a local view of the system, in a finite amount of time the system converges to a global state with desired property. In a graph G = (V, E), a subset S C V is a 2-packing if Vi c V: IN[i] n SI <1. In this paper, an ID-based, constant space, self-stabilizing algorithm that stabilizes to a maximal 2-packing in an arbitrary graph is proposed. It is shown that the algorithm stabilizes in 0(n3) moves under any scheduler (daemon). Specifically, it is shown that the algorithm stabilizes in linear time-steps under a synchronous daemon where every privileged node moves at each time-step.

Keywords: self-stabilization, 2-packing, distributed computing, fault tolerance, graph algorithms

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Authors: Francisco Bulnes

Abstract:

Through the Fukaya conjecture and the wrapped Floer cohomology, the correspondences between paths in a loop space and states of a wrapping space of states in a Hamiltonian space (the ramification of field in this case is the connection to the operator that goes from TM to T*M) are demonstrated where these last states are corresponding to bosonic extensions of a spectrum of the space-time or direct image of the functor Spec, on space-time. This establishes a distinguished diffeomorphism defined by the mapping from the corresponding loops space to wrapping category of the Floer cohomology complex which furthermore relates in certain proportion D-branes (certain D-modules) with strings. This also gives to place to certain conjecture that establishes equivalences between moduli spaces that can be consigned in a moduli identity taking as space-time the Hitchin moduli space on G, whose dual can be expressed by a factor of a bosonic moduli spaces.

Keywords: Floer cohomology, Fukaya conjecture, Lagrangian submanifolds, spectrum of ring, topological quantum diffeomorphisms.

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Authors: Mohammad Bakhshi, Firas Al Janabi

Abstract:

High resolution rain data are very important to fulfill the input of hydrological models. Among models of high-resolution rainfall data generation, the temporal disaggregation was chosen for this study. The paper attempts to generate three different rainfall resolutions (4-hourly, hourly and 10-minutes) from daily for around 20-year record period. The process was done by DiMoN tool which is based on random cascade model and method of fragment. Differences between observed and simulated rain dataset are evaluated with variety of statistical and empirical methods: Kolmogorov-Smirnov test (K-S), usual statistics, and Exceedance probability. The tool worked well at preserving the daily rainfall values in wet days, however, the generated data are cumulated in a shorter time period and made stronger storms. It is demonstrated that the difference between generated and observed cumulative distribution function curve of 4-hourly datasets is passed the K-S test criteria while in hourly and 10-minutes datasets the P-value should be employed to prove that their differences were reasonable. The results are encouraging considering the overestimation of generated high-resolution rainfall data.

Keywords: DiMoN tool, disaggregation, exceedance probability, Kolmogorov-Smirnov Test, rainfall.

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Authors: Dong-Woo Lim, Chang-Jae Chun, Hyung-Myung Kim

Abstract:

We consider a two-way relay network where two sources exchange information. A relay helps the two sources exchange information using the decode-and-XOR-forward protocol. We investigate the power minimization problem with minimum rate constraints. The system needs two time slots and in each time slot the required rate pair should be achievable. The power consumption is minimized in each time slot and we obtained the closed form solution. The simulation results confirm that the proposed power allocation scheme consumes lower total power than the conventional schemes.

Keywords: Decode-and-XOR-forward, power minimization, two-way relay

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Authors: Tamás Hartványi, Ferenc Tóth

Abstract:

European telecommunication distribution center performance is measured by service lead time and quality. Operation model is CTO (customized to order) namely, a high mix customization of telecommunication network equipment and parts. CTO operation contains material receiving, warehousing, network and server assembly to order and configure based on customer specifications. Variety of the product and orders does not support mass production structure. One of the success factors to satisfy customer is to have a proper aggregated planning method for the operation in order to have optimized human resources and highly efficient asset utilization. Research will investigate several methods and find proper way to have an order book simulation where practical optimization problem may contain thousands of variables and the simulation running times of developed algorithms were taken into account with high importance. There are two operation research models that were developed, customer demand is given in orders, no change over time, customer demands are given for product types, and changeover time is constant.

Keywords: CTO, aggregated planning, demand simulation, changeover time.

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Authors: Lili Wang, Meng Hu

Abstract:

In this paper, a class of recurrent neural networks (RNNs) with variable delays are studied on almost periodic time scales, some sufficient conditions are established for the existence and global exponential stability of the almost periodic solution. These results have important leading significance in designs and applications of RNNs. Finally, two examples and numerical simulations are presented to illustrate the feasibility and effectiveness of the results.

Keywords: Recurrent neural network, Almost periodic solution, Global exponential stability, Time scale.

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Authors: Jing Zhang, Jie Chen

Abstract:

In this paper, we mainly analyze an automated parking system where the storage and retrieval requests are performed by a tower crane. In this parking system, the S/R crane which is located at the middle of the bottom of the cylinder parking area can rotate in both clockwise and counterclockwise and three kinds of movements can be done simultaneously. We develop some mathematical travel time models for the single command cycle under the random storage assignment using the characteristics of this system. Finally, we compare these travel models with discrete case and it is shown that these travel models display a good satisfactory performance.

Keywords: Parking system, travel time model, tower crane.

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Authors: M. K. Benhaoua, A. K. Singh, A. E. H. Benyamina, A. Kumar, P. Boulet

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In this paper, we propose a new packing strategy to find a free resource for run-time mapping of application tasks to NoC-based Heterogeneous MPSoC. The proposed strategy minimizes the task mapping time in addition to placing the communicating tasks close to each other. To evaluate our approach, a comparative study is carried out for a platform containing single task supported PEs. Experiments show that our strategy provides better results when compared to latest dynamic mapping strategies reported in the literature.

Keywords: Multi-Processor Systems-on-Chip (MPSoCs), Network-on-Chip (NoC), Heterogeneous architectures, Dynamic mapping heuristics.

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Authors: Gerrit Alves, Jürgen Roßmann, Roland Wischnewski

Abstract:

Today, transport and logistic systems are often tightly integrated in the production. Lean production and just-in-time delivering create multiple constraints that have to be fulfilled. As transport networks often have evolved over time they are very expensive to change. This paper describes a discrete-event-simulation system which simulates transportation models using real time resource routing and collision avoidance. It allows for the specification of own control algorithms and validation of new strategies. The simulation is integrated into a virtual reality (VR) environment and can be displayed in 3-D to show the progress. Simulation elements can be selected through VR metaphors. All data gathered during the simulation can be presented as a detailed summary afterwards. The included cost-benefit calculation can help to optimize the financial outcome. The operation of this approach is shown by the example of a timber harvest simulation.

Keywords: Discrete-Event-Simulation, Logistic, Simulation, Virtual Reality.

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Authors: Aaminah Hassan

Abstract:

Advertisements carry a great potential to influence our lives because they are crafted to meet particular ends. Stereotypical representation in advertisements is capable of forming unconscious attitudes among people towards any gender and their abilities. This study focuses on gender representation in Pakistani prime time advertisements. For this purpose, 13 advertisements were selected from three different categories of foods and beverages, cosmetics, cell phones and cellular networks from the prime time slots of one of the leading Pakistani entertainment channel, ‘Urdu 1’. Both quantitative and qualitative analyses are carried out for range of variables like gender, age, roles, activities, setting, appearance and voice overs. The results revealed that gender representation in advertisements is stereotypical. Moreover, in few instances, the portrayal of women is not only culturally inappropriate but is demeaning to the image of women as well. Their bodily charm is used to promote products. Comparing different entertainment channels for their prime time advertisements and broadening the scope of this research will yield greater implications for the researchers who want to carry out the similar research. It is hoped that the current study would help in the promotion of media literacy among the viewers and media authorities in Pakistan.

Keywords: Advertisements, content analysis, gender, prime time.

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Authors: B. S. Raghavendra, D. Narayana Dutt

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In this paper, we have developed a method to compute fractal dimension (FD) of discrete time signals, in the time domain, by modifying the box-counting method. The size of the box is dependent on the sampling frequency of the signal. The number of boxes required to completely cover the signal are obtained at multiple time resolutions. The time resolutions are made coarse by decimating the signal. The loglog plot of total number of boxes required to cover the curve versus size of the box used appears to be a straight line, whose slope is taken as an estimate of FD of the signal. The results are provided to demonstrate the performance of the proposed method using parametric fractal signals. The estimation accuracy of the method is compared with that of Katz, Sevcik, and Higuchi methods. In addition, some properties of the FD are discussed.

Keywords: Box-counting, Fractal dimension, Higuchi method, Katz method, Parametric fractal signals, Sevcik method.

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Authors: Mehrdad Hashemi, Mitra Behrooz Aghdam, Reza Mahdian, Ahmad Reza Kamyab

Abstract:

Cytogenetic analysis still remains the gold standard method for prenatal diagnosis of trisomy 21 (Down syndrome, DS). Nevertheless, the conventional cytogenetic analysis needs live cultured cells and is too time-consuming for clinical application. In contrast, molecular methods such as FISH, QF-PCR, MLPA and quantitative Real-time PCR are rapid assays with results available in 24h. In the present study, we have successfully used a novel MGB TaqMan probe-based real time PCR assay for rapid diagnosis of trisomy 21 status in Down syndrome samples. We have also compared the results of this molecular method with corresponding results obtained by the cytogenetic analysis. Blood samples obtained from DS patients (n=25) and normal controls (n=20) were tested by quantitative Real-time PCR in parallel to standard G-banding analysis. Genomic DNA was extracted from peripheral blood lymphocytes. A high precision TaqMan probe quantitative Real-time PCR assay was developed to determine the gene dosage of DSCAM (target gene on 21q22.2) relative to PMP22 (reference gene on 17p11.2). The DSCAM/PMP22 ratio was calculated according to the formula; ratio=2 -ΔΔCT. The quantitative Real-time PCR was able to distinguish between trisomy 21 samples and normal controls with the gene ratios of 1.49±0.13 and 1.03±0.04 respectively (p value <0.001). These results represent the presence of 3 copies of target gene in DS samples Vs 2 copies in normal controls. The results of quantitative Real-time PCR were in complete agreement with results of cytogenetic analysis. This study confirms previous reports regarding successful implementation of quantitative Real-time PCR for detection of trisomy 21. However, the assay has been improved by using MGB probes and more accurate data analysis. This assay, in particular, when performed in combination with another molecular assay such as QF-PCR or MLPA, can be used as a reliable technique for rapid prenatal diagnosis of trisomy 21.

Keywords: Trisomy 21, Real-time PCR, MGB-TaqMan Probes, Gene Dosage.

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