Search results for: Modified Synchronous Detection Method

Authors: J. Madureira, R. Lagido, I. Sousa

Abstract:

Surf is an increasingly popular sport and its performance evaluation is often qualitative. This work aims at using a smartphone to collect and analyze the GPS and inertial sensors data in order to obtain quantitative metrics of the surfing performance. Two approaches are compared for detection of wave rides, computing the number of waves rode in a surfing session, the starting time of each wave and its duration. The first approach is based on computing the velocity from the Global Positioning System (GPS) signal and finding the velocity thresholds that allow identifying the start and end of each wave ride. The second approach adds information from the Inertial Measurement Unit (IMU) of the smartphone, to the velocity thresholds obtained from the GPS unit, to determine the start and end of each wave ride. The two methods were evaluated using GPS and IMU data from two surfing sessions and validated with similar metrics extracted from video data collected from the beach. The second method, combining GPS and IMU data, was found to be more accurate in determining the number of waves, start time and duration. This paper shows that it is feasible to use smartphones for quantification of performance metrics during surfing. In particular, detection of the waves rode and their duration can be accurately determined using the smartphone GPS and IMU. 

Keywords: Inertial Measurement Unit (IMU), Global Positioning System (GPS), smartphone, surfing performance.

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Authors: Luciano Nieddu, Giuseppe Manfredi, Salvatore D'Acunto, Katia La Regina

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In this paper a non-parametric statistical pattern recognition algorithm for the problem of credit scoring will be presented. The proposed algorithm is based on a clustering k- means algorithm and allows for the determination of subclasses of homogenous elements in the data. The algorithm will be tested on two benchmark datasets and its performance compared with other well known pattern recognition algorithm for credit scoring.

Keywords: Constrained clustering, Credit scoring, Statistical pattern recognition, Supervised classification.

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Authors: Taiwo. M. Akinmuyisitan, John Cosmas

Abstract:

This paper practically demonstrates the theories and concepts of an Advanced Convolutional Neural Network in the design and development of a scalable artificial intelligence model for the detection of criminal masterminds. The technique uses machine vision algorithms to compute the facial characteristics of suspects and classify actors as criminal or non-criminal faces. The paper proceeds further to compare the results of the error accuracy of two popular custom convolutional pre-trained networks, VGG16 and Resnet50. The result shows that VGG16 is probably more efficient than ResNet50 for the dataset we used.

Keywords: Artificial intelligence, convolutional neural networks, Resnet50, VGG16.

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Authors: Shunya Wakayama, Kazuya Okubo, Toru Fujii, Daisuke Sakata, Noriyuki Kado, Hiroshi Furutachi

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The purpose of this study is to propose an effective method to improve frictional coefficient between shoe rubber soles with added glass fibers and the surfaces of icy and snowy road in order to prevent slip-and-fall accidents by the users. The additional fibers into the rubber were uniformly tilted to the perpendicular direction of the frictional surface, where tilting angles were -60, -30, +30, +60, 90 degrees and 0 (as normal specimen), respectively. It was found that parallel arraignment was effective to improve the frictional coefficient when glass fibers were embedded in the shoe rubber, while perpendicular to normal direction of the embedded glass fibers on the shoe surface was also effective to do that once after they were exposed from the shoe rubber with its abrasion. These improvements were explained by the increase of stiffness against the shear deformation of the rubber at critical frictional state and adequate scratching of fibers when fibers were protruded in perpendicular to frictional direction, respectively. Most effective angle of tilting of frictional coefficient between rubber specimens and a stone was perpendicular (= 0 degree) to frictional direction. Combinative modified rubber specimen having 2 layers was fabricated where tilting angle of protruded fibers was 0 degree near the contact surface and tilting angle of embedded fibers was 90 degrees near back surface in thickness direction to further improve the frictional coefficient. Current study suggested that effective arraignments in tilting angle of the added fibers should be applied in designing rubber shoe soles to keep the safeties for users in regions of cold climates.

Keywords: Frictional coefficient, icy and snowy road, shoe rubber soles, tilting angle.

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Authors: Po-Chun Lin, Jyh-Cherng Gu

Abstract:

This study adopted previous fault patterns, results of detection analysis, historical records and data, and experts- experiences to establish fuzzy principles and estimate the failure probability index of components of a power transformer. Considering that actual parameters and limiting conditions of parameters may differ, this study used the standard data of IEC, IEEE, and CIGRE as condition parameters. According to the characteristics of each condition parameter, relative degradation was introduced to reflect the degree of influence of the factors on the transformer condition. The method of fuzzy mathematics was adopted to determine the subordinate function of the transformer condition. The calculation used the Matlab Fuzzy Tool Box to select the condition parameters of coil winding, iron core, bushing, OLTC, insulating oil and other auxiliary components and factors (e.g., load records, performance history, and maintenance records) of the transformer to establish the fuzzy principles. Examples were presented to support the rationality and effectiveness of the evaluation method of power transformer performance conditions, as based on fuzzy comprehensive evaluation.

Keywords: Fuzzy, relative degradation degree, condition-basedmaintenance, power transformer

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Authors: Noor Ayuni Che Zakaria, Takashi Komeda, Cheng Yee Low, Kaoru Inoue, Fazah Akhtar Hanapiah

Abstract:

Previous studies have shown that there are arguments regarding the reliability and validity of the Ashworth and Modified Ashworth Scale towards evaluating patients diagnosed with upper limb disorders. These evaluations depended on the raters’ experiences. This initiated us to develop an upper limb disorder part-task trainer that is able to simulate consistent upper limb disorders, such as spasticity and rigidity signs, based on the Modified Ashworth Scale to improve the variability occurring between raters and intra-raters themselves. By providing consistent signs, novice therapists would be able to increase training frequency and exposure towards various levels of signs. A total of 22 physiotherapists and occupational therapists participated in the study. The majority of the therapists agreed that with current therapy education, they still face problems with inter-raters and intra-raters variability (strongly agree 54%; n = 12/22, agree 27%; n = 6/22) in evaluating patients’ conditions. The therapists strongly agreed (72%; n = 16/22) that therapy trainees needed to increase their frequency of training; therefore believe that our initiative to develop an upper limb disorder training tool will help in improving the clinical education field (strongly agree and agree 63%; n = 14/22).

Keywords: Upper limb disorders, Clinical education tool, Inter/intra-raters variability, Spasticity, Modified Ashworth Scale.

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Authors: J. G. Batista, K. N. Sousa, J. L. Nunes, R. L. S. Sousa, G. A. P. Thé

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This work deals with parameter identification of permanent magnet motors, a class of ac motor which is particularly important in industrial automation due to characteristics like applications high performance, are very attractive for applications with limited space and reducing the need to eliminate because they have reduced size and volume and can operate in a wide speed range, without independent ventilation. By using experimental data and genetic algorithm we have been able to extract values for both the motor inductance and the electromechanical coupling constant, which are then compared to measured and/or expected values.

Keywords: Modeling, AC servomotor, Permanent Magnet Synchronous Motor-PMSM, Genetic Algorithm, Vector Control, Robotic Manipulator, Control.

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Authors: Sheema Shuja Khattak, Gule Saman, Imran Khan, Abdus Salam

Abstract:

Image segmentation plays an important role in medical imaging applications. Therefore, accurate methods are needed for the successful segmentation of medical images for diagnosis and detection of various diseases. In this paper, we have used maximum entropy to achieve image segmentation. Maximum entropy has been calculated using Shannon, Renyi and Tsallis entropies. This work has novelty based on the detection of skin lesion caused by the bite of a parasite called Sand Fly causing the disease is called Cutaneous Leishmaniasis.

Keywords: Shannon, Maximum entropy, Renyi, Tsallis entropy.

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Authors: M. Das Gupta, S. Banerjee

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Content Based Image Retrieval (CBIR) coupled with Case Based Reasoning (CBR) is a paradigm that is becoming increasingly popular in the diagnosis and therapy planning of medical ailments utilizing the digital content of medical images. This paper presents a survey of some of the promising approaches used in the detection of abnormalities in retina images as well in mammographic screening and detection of regions of interest in MRI scans of the brain. We also describe our proposed algorithm to detect hard exudates in fundus images of the retina of Diabetic Retinopathy patients.

Keywords: Case based reasoning, Exudates, Retina image, Similarity based retrieval.

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Authors: M. Kovac

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This article describes design of the 8-bit asynchronous microcontroller simulation model in VHDL. The model is created in ISE Foundation design tool and simulated in Modelsim tool. This model is a simple application example of asynchronous systems designed in synchronous design tools. The design process of creating asynchronous system with 4-phase bundled-data protocol and with matching delays is described in the article. The model is described in gate-level abstraction. The simulation waveform of the functional construction is the result of this article. Described construction covers only the simulation model. The next step would be creating synthesizable model to FPGA.

Keywords: Asynchronous, Microcontroller, VHDL, FPGA.

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Authors: J.S.C. Prentice

Abstract:

The RK1GL2X3 method is a numerical method for solving initial value problems in ordinary differential equations, and is based on the RK1GL2 method which, in turn, is a particular case of the general RKrGLm method. The RK1GL2X3 method is a fourth-order method, even though its underlying Runge-Kutta method RK1 is the first-order Euler method, and hence, RK1GL2X3 is considerably more efficient than RK1. This enhancement is achieved through an implementation involving triple-nested two-point Gauss- Legendre quadrature.

Keywords: RK1GL2X3, RK1GL2, RKrGLm, Runge-Kutta, Gauss-Legendre, initial value problem, local error, global error.

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Authors: Mahdi Pirmoradian, Olayinka Adigun, Christos Politis

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Cooperative spectrum sensing is a crucial challenge in cognitive radio networks. Cooperative sensing can increase the reliability of spectrum hole detection, optimize sensing time and reduce delay in cooperative networks. In this paper, an efficient central capacity optimization algorithm is proposed to minimize cooperative sensing time in a homogenous sensor network using OR decision rule subject to the detection and false alarm probabilities constraints. The evaluation results reveal significant improvement in the sensing time and normalized capacity of the cognitive sensors.

Keywords: Cooperative networks, normalized capacity, sensing time.

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Authors: V. Venkatachalam, S. Selvan

Abstract:

The security of computer networks plays a strategic role in modern computer systems. Intrusion Detection Systems (IDS) act as the 'second line of defense' placed inside a protected network, looking for known or potential threats in network traffic and/or audit data recorded by hosts. We developed an Intrusion Detection System using LAMSTAR neural network to learn patterns of normal and intrusive activities, to classify observed system activities and compared the performance of LAMSTAR IDS with other classification techniques using 5 classes of KDDCup99 data. LAMSAR IDS gives better performance at the cost of high Computational complexity, Training time and Testing time, when compared to other classification techniques (Binary Tree classifier, RBF classifier, Gaussian Mixture classifier). we further reduced the Computational Complexity of LAMSTAR IDS by reducing the dimension of the data using principal component analysis which in turn reduces the training and testing time with almost the same performance.

Keywords: Binary Tree Classifier, Gaussian Mixture, IntrusionDetection System, LAMSTAR, Radial Basis Function.

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Authors: A. Tellaeche, X. P. Burgos-Artizzu, G. Pajares, A. Ribeiro

Abstract:

One important objective in Precision Agriculture is to minimize the volume of herbicides that are applied to the fields through the use of site-specific weed management systems. In order to reach this goal, two major factors need to be considered: 1) the similar spectral signature, shape and texture between weeds and crops; 2) the irregular distribution of the weeds within the crop's field. This paper outlines an automatic computer vision system for the detection and differential spraying of Avena sterilis, a noxious weed growing in cereal crops. The proposed system involves two processes: image segmentation and decision making. Image segmentation combines basic suitable image processing techniques in order to extract cells from the image as the low level units. Each cell is described by two area-based attributes measuring the relations among the crops and the weeds. From these attributes, a hybrid decision making approach determines if a cell must be or not sprayed. The hybrid approach uses the Support Vector Machines and the Fuzzy k-Means methods, combined through the fuzzy aggregation theory. This makes the main finding of this paper. The method performance is compared against other available strategies.

Keywords: Fuzzy k-Means, Precision agriculture, SupportVectors Machines, Weed detection.

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Authors: Venny, S. Gan, H. K. Ng

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This work focuses on the remediation of polycyclic aromatic hydrocarbons (PAHs)-contaminated soil via Fenton treatment coupled with novel chelating agent (CA). The feasibility of chelated modified Fenton (MF) treatment to promote PAH oxidation in artificially contaminated soils was investigated in laboratory scale batch experiments at natural pH. The effects of adding inorganic and organic CA are discussed. Experiments using different iron catalyst to CA ratios were conducted, resulting in hydrogen peroxide: soil: iron: CA weight ratios that varied from 0.049: 1: 0.072: 0.008 to 0.049: 1: 0.072: 0.067. The results revealed that (1) inorganic CA could provide much higher PAH removal efficiency and (2) most of the proposed CAs were more efficient than commonly utilised CAs even at mild ratio. This work highlights the potential of novel chelating agents in maintaining a suitable environment throughout the Fenton treatment, particularly in soils with high buffer capacity.

Keywords: Chelating agent, Fenton, hydroxyl radicals, polycyclic aromatic hydrocarbon.

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Authors: Wenjuan Du, Jingtian Bi, Tong Wang, Haifeng Wang

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Grid connection of a large-scale wind farm affects power system small-signal angular stability in two aspects. Firstly, connection of the wind farm brings about the change of load flow and configuration of a power system. Secondly, the dynamic interaction is introduced by the wind farm with the synchronous generators (SGs) in the power system. This paper proposes a method to assess the two aspects of the effect of the wind farm on power system small-signal angular stability. The effect of the change of load flow/system configuration brought about by the wind farm can be examined separately by displacing wind farms with constant power sources, then the effect of the dynamic interaction of the wind farm with the SGs can be also computed individually. Thus, a clearer picture and better understanding on the power system small-signal angular stability as affected by grid connection of the large-scale wind farm are provided. In the paper, an example power system with grid connection of a wind farm is presented to demonstrate the proposed approach.

Keywords: power system small-signal angular stability, power system low-frequency oscillations, electromechanical oscillation modes, wind farms, double fed induction generator (DFIG)

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Authors: Jinming Ma, Tianbing Xia, Janusz R. Getta

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This paper presents the application of fuzzy set theory to implement of mobile advertisement anti-fraud systems. Mobile anti-fraud is a method aiming to identify mobile advertisement fraudsters. One of the main problems of mobile anti-fraud is the lack of evidence to prove a user to be a fraudster. In this paper, we implement an application by using fuzzy set theory to demonstrate how to detect cheaters. The advantage of our method is that the hardship in detecting fraudsters in small data samples has been avoided. We achieved this by giving each user a suspicious degree showing how likely the user is cheating and decide whether a group of users (like all users of a certain APP) together to be fraudsters according to the average suspicious degree. This makes the process more accurate as the data of a single user is too small to be predictable.

Keywords: Mobile internet, advertisement, anti-fraud, fuzzy set theory.

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Authors: Yanping Liao, Congcong He, Ruigang Zhao

Abstract:

The application of magnetocardiography signals to detect cardiac electrical function is a new technology developed in recent years. The magnetocardiography signal is detected with Superconducting Quantum Interference Devices (SQUID) and has considerable advantages over electrocardiography (ECG). It is difficult to extract Magnetocardiography (MCG) signal which is buried in the noise, which is a critical issue to be resolved in cardiac monitoring system and MCG applications. In order to remove the severe background noise, the Total Variation (TV) regularization method is proposed to denoise MCG signal. The approach transforms the denoising problem into a minimization optimization problem and the Majorization-minimization algorithm is applied to iteratively solve the minimization problem. However, traditional TV regularization method tends to cause step effect and lacks constraint adaptability. In this paper, an improved TV regularization method for denoising MCG signal is proposed to improve the denoising precision. The improvement of this method is mainly divided into three parts. First, high-order TV is applied to reduce the step effect, and the corresponding second derivative matrix is used to substitute the first order. Then, the positions of the non-zero elements in the second order derivative matrix are determined based on the peak positions that are detected by the detection window. Finally, adaptive constraint parameters are defined to eliminate noises and preserve signal peak characteristics. Theoretical analysis and experimental results show that this algorithm can effectively improve the output signal-to-noise ratio and has superior performance.

Keywords: Constraint parameters, derivative matrix, magnetocardiography, regular term, total variation.

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Authors: Akram A. Mustafa

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In this paper an approaches for increasing the effectiveness of error detection in computer network channels with Pulse-Amplitude Modulation (PAM) has been proposed. Proposed approaches are based on consideration of special feature of errors, which are appearances in line with PAM. The first approach consists of CRC modification specifically for line with PAM. The second approach is base of weighted checksums using. The way for checksum components coding has been developed. It has been shown that proposed checksum modification ensure superior digital data control transformation reliability for channels with PAM in compare to CRC.

Keywords: Pulse-Amplitude Modulation, checksum, transmission, discrete.

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Authors: Mohammed Salem Alzahrani

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In this paper the optimality of the solution of an existing real word assignment problem known as the seat assignment problem using Seat Assignment Method (SAM) is discussed. SAM is the newly driven method from three existing methods, Hungarian Method, Northwest Corner Method and Least Cost Method in a special way that produces the easiness & fairness among all methods that solve the seat assignment problem.

Keywords: Assignment Problem, Hungarian Method, Least Cost Method, Northwest Corner Method, Seat Assignment Method (SAM), A Real Word Assignment Problem.

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Authors: Raymond Xu, Ashley Hua, Andrew Wang, Yuru Lin

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During COVID-19, the depression rate has increased dramatically. Young adults are most vulnerable to the mental health effects of the pandemic. Lower-income families have a higher ratio to be diagnosed with depression than the general population, but less access to clinics. This research aims to achieve early depression detection at low cost, large scale, and high accuracy with an interdisciplinary approach by incorporating clinical practices defined by American Psychiatric Association (APA) as well as multimodal AI framework. The proposed approach detected the nine depression symptoms with Natural Language Processing sentiment analysis and a symptom-based Lexicon uniquely designed for young adults. The experiments were conducted on the multimedia survey results from adolescents and young adults and unbiased Twitter communications. The result was further aggregated with the facial emotional cues analyzed by the Convolutional Neural Network on the multimedia survey videos. Five experiments each conducted on 10k data entries reached consistent results with an average accuracy of 88.31%, higher than the existing natural language analysis models. This approach can reach 300+ million daily active Twitter users and is highly accessible by low-income populations to promote early depression detection to raise awareness in adolescents and young adults and reveal complementary cues to assist clinical depression diagnosis.

Keywords: Artificial intelligence, depression detection, facial emotion recognition, natural language processing, mental disorder.

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Authors: Tze Sian Pui, Tushar Bansal, Patthara Kongsuphol, Sunil K. Arya

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We present a label-free biosensor based on electrochemical impedance spectroscopy for the detection of proinflammatory cytokine Tumor Necrosis Factor (TNF-α). Secretion of TNF-α has been correlated to the onset of various diseases including rheumatoid arthritis, Crohn-s disease etc. Gold electrodes were patterned on a silicon substrate and self assembled monolayer of dithiobis-succinimidyl propionate was used to develop the biosensor which achieved a detection limit of ~57fM. A linear relationship was also observed between increasing TNF-α concentrations and chargetransfer resistance within a dynamic range of 1pg/ml – 1ng/ml.

Keywords: Tumor necrosis factor, electrochemical impedance spectroscopy, label free, self assembled monolayer

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Authors: S. Sowmyayani, P. Arockia Jansi Rani

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This paper integrates Octagon and Square Search pattern (OCTSS) motion estimation algorithm into H.264/AVC (Advanced Video Coding) video codec in Adaptive Group of Pictures (AGOP) mode. AGOP structure is computed based on scene change in the video sequence. Octagon and square search pattern block-based motion estimation method is implemented in inter-prediction process of H.264/AVC. Both these methods reduce bit rate and computational complexity while maintaining the quality of the video sequence respectively. Experiments are conducted for different types of video sequence. The results substantially proved that the bit rate, computation time and PSNR gain achieved by the proposed method is better than the existing H.264/AVC with fixed GOP and AGOP. With a marginal gain in quality of 0.28dB and average gain in bitrate of 132.87kbps, the proposed method reduces the average computation time by 27.31 minutes when compared to the existing state-of-art H.264/AVC video codec.

Keywords: Block Distortion Measure, Block Matching Algorithms, H.264/AVC, Motion estimation, Search patterns, Shot cut detection.

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Authors: Wiriya Loongyai, Kiettisak Promphet, Nilubol Kangsukul, Ratchawat Noppha

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Polymerase chain reaction (PCR) assay and conventional microbiological methods were used to detect bacterial contamination of egg shells and egg content in different commercial housing systems, open house system and evaporative cooling system. A PCR assay was developed for direct detection using a set of primers specific for the invasion by A gene (invA) of Salmonella spp. PCR detected the presence of Salmonella in 2 samples of shell egg from the evaporative cooling system, while conventional cultural methods detected no Salmonella from the same samples.

Keywords: egg content, egg shell, invA gene, PCR, Salmonellaspp.

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Authors: Fatima Zohra Mahi, Luca Varani

Abstract:

We present an analytical model for the calculation of the sensitivity, the spectral current noise and the detective parameter for an optically illuminated In0.53Ga0.47As n+nn+ diode. The photocurrent due to the excess carrier is obtained by solving the continuity equation. Moreover, the current noise level is evaluated at room temperature and under a constant voltage applied between the diode terminals. The analytical calculation of the current noise in the n+nn+ structure is developed by considering the free carries fluctuations. The responsivity and the detection parameter are discussed as functions of the doping concentrations and the emitter layer thickness in one-dimensional homogeneous n+nn+ structure.

Keywords: Responsivity, detection parameter, photo-detectors, continuity equation, current noise.

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Authors: Roberto Sabatini, Alessandro Gardi, Mark A. Richardson

Abstract:

The availability of powerful eye-safe laser sources and the recent advancements in electro-optical and mechanical beam-steering components have allowed laser-based Light Detection and Ranging (LIDAR) to become a promising technology for obstacle warning and avoidance in a variety of manned and unmanned aircraft applications. LIDAR outstanding angular resolution and accuracy characteristics are coupled to its good detection performance in a wide range of incidence angles and weather conditions, providing an ideal obstacle avoidance solution, which is especially attractive in low-level flying platforms such as helicopters and small-to-medium size Unmanned Aircraft (UA). The Laser Obstacle Avoidance Marconi (LOAM) system is one of such systems, which was jointly developed and tested by SELEX-ES and the Italian Air Force Research and Flight Test Centre. The system was originally conceived for military rotorcraft platforms and, in this paper, we briefly review the previous work and discuss in more details some of the key development activities required for integration of LOAM on UA platforms. The main hardware and software design features of this LOAM variant are presented, including a brief description of the system interfaces and sensor characteristics, together with the system performance models and data processing algorithms for obstacle detection, classification and avoidance. In particular, the paper focuses on the algorithm proposed for optimal avoidance trajectory generation in UA applications.

Keywords: LIDAR, Low-Level Flight, Nap-of-the-Earth Flight, Near Infra-Red, Obstacle Avoidance, Obstacle Detection, Obstacle Warning System, Sense and Avoid, Trajectory Optimisation, Unmanned Aircraft.

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Authors: Yong-Gyu Lee, Gilwon Yoon

Abstract:

Bleeding in the digestive duct is an important diagnostic parameter for patients. Blood in the endoscopic image can be determined by investigating the color tone of blood due to the degree of oxygenation, under- or over- illumination, food debris and secretions, etc. However, we found that how to pre-process raw images obtained from the capsule detectors was very important. We applied various image process methods suitable for the capsule endoscopic image in order to remove noises and unbalanced sensitivities for the image pixels. The results showed that much improvement was achieved by additional pre-processing techniques on the algorithm of determining bleeding areas.

Keywords: blood detection, capsule endoscopy, image processing.

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Authors: Yuki. Akiyama, Hiroaki. Sengoku, Ryosuke. Shibasaki

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In this study, the locations and areas of commercial accumulations were detected by using digital yellow page data. An original buffering method that can accurately create polygons of commercial accumulations is proposed in this paper.; by using this method, distribution of commercial accumulations can be easily created and monitored over a wide area. The locations, areas, and time-series changes of commercial accumulations in the South Kanto region can be monitored by integrating polygons of commercial accumulations with the time-series data of digital yellow page data. The circumstances of commercial accumulations were shown to vary according to areas, that is, highly- urbanized regions such as the city center of Tokyo and prefectural capitals, suburban areas near large cities, and suburban and rural areas.

Keywords: Commercial accumulations, Spatio-temporal analysis, Urban monitoring, Yellow page data

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Authors: Srinivas Bathini, Duraichelvan Raju, Simona Badilescu, Muthukumaran Packirisamy

Abstract:

A bio-sensing method, based on the plasmonic property of gold nano-islands, has been developed for detection of exosomes in a clinical setting. The position of the gold plasmon band in the UV-Visible spectrum depends on the size and shape of gold nanoparticles as well as on the surrounding environment. By adsorbing various chemical entities, or binding them, the gold plasmon band will shift toward longer wavelengths and the shift is proportional to the concentration. Exosomes transport cargoes of molecules and genetic materials to proximal and distal cells. Presently, the standard method for their isolation and quantification from body fluids is by ultracentrifugation, not a practical method to be implemented in a clinical setting. Thus, a versatile and cutting-edge platform is required to selectively detect and isolate exosomes for further analysis at clinical level. The new sensing protocol, instead of antibodies, makes use of a specially synthesized polypeptide (Vn96), to capture and quantify the exosomes from different media, by binding the heat shock proteins from exosomes. The protocol has been established and optimized by using a glass substrate, in order to facilitate the next stage, namely the transfer of the protocol to a microfluidic environment. After each step of the protocol, the UV-Vis spectrum was recorded and the position of gold Localized Surface Plasmon Resonance (LSPR) band was measured. The sensing process was modelled, taking into account the characteristics of the nano-island structure, prepared by thermal convection and annealing. The optimal molar ratios of the most important chemical entities, involved in the detection of exosomes were calculated as well. Indeed, it was found that the results of the sensing process depend on the two major steps: the molar ratios of streptavidin to biotin-PEG-Vn96 and, the final step, the capture of exosomes by the biotin-PEG-Vn96 complex. The microfluidic device designed for sensing of exosomes consists of a glass substrate, sealed by a PDMS layer that contains the channel and a collecting chamber. In the device, the solutions of linker, cross-linker, etc., are pumped over the gold nano-islands and an Ocean Optics spectrometer is used to measure the position of the Au plasmon band at each step of the sensing. The experiments have shown that the shift of the Au LSPR band is proportional to the concentration of exosomes and, thereby, exosomes can be accurately quantified. An important advantage of the method is the ability to discriminate between exosomes having different origins.

Keywords: Exosomes, gold nano-islands, microfluidics, plasmonic biosensing.

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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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