921 resultados para Docker,ARM,Raspberry PI,single board computer,QEMU,Sabayon Linux,Gentoo Linux
Resumo:
Extensive high-grade polydeformed metamorphic provinces surrounding Archaean cratonic nuclei in the East Antarctic Shield record two tectono-thermal episodes in late Mesoproterozoic and late Neoproterozoic-Cambrian times. In Western Dronning Maud Land, the high-grade Mesoproterozoic Maud Belt is juxtaposed against the Archaean Grunehogna Province and has traditionally been interpreted as a Grenvillian mobile belt that was thermally overprinted during the Early Palaeozoic. Integration of new U-Pb sensitive high-resolution ion microprobe and conventional single zircon and monazite age data, and Ar-Ar data on hornblende and biotite, with thermobarometric calculations on rocks from the H.U. Sverdrupfjella, northern Maud Belt, resulted in a more complex P-T-t evolution than previously assumed. A c. 540?Ma monazite, hosted by an upper ampibolite-facies mineral assemblage defining a regionally dominant top-to-NW shear fabric, provides strong evidence for the penetrative deformation in the area being of Pan-African age and not of Grenvillian age as previously reported. Relics of an eclogite-facies garnet-omphacite assemblage within strain-protected mafic boudins indicate that the peak metamorphic conditions recorded by most rocks in the area (T = 687-758°C, P = 9·4-11·3?kbar) were attained subsequent to decompression from P > 12·9?kbar. By analogy with limited U-Pb single zircon age data and on circumstantial textural grounds, this earlier eclogite-facies metamorphism is ascribed to subduction and accretion around 565?Ma. Post-peak metamorphic K-metasomatism under amphibolite-facies conditions is ascribed to the intrusion of post-orogenic granite at c. 480?Ma. The recognition of extensive Pan-African tectonism in the Maud Belt casts doubts on previous Rodinia reconstructions, in which this belt takes a pivotal position between East Antarctica, the Kalahari Craton and Laurentia. Evidence of late Mesoproterozoic high-grade metamorphism during the formation of the Maud Belt exists in the form of c. 1035?Ma zircon overgrowths that are probably related to relics of granulite-facies metamorphism recorded from other parts of the Maud Belt. The polymetamorphic rocks are largely derived from a c. 1140?Ma volcanic arc and 1072 ± 10?Ma granite.
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Single core capabilities have reached their maximum clock speed; new multicore architectures provide an alternative way to tackle this issue instead. The design of decoding applications running on top of these multicore platforms and their optimization to exploit all system computational power is crucial to obtain best results. Since the development at the integration level of printed circuit boards are increasingly difficult to optimize due to physical constraints and the inherent increase in power consumption, development of multiprocessor architectures is becoming the new Holy Grail. In this sense, it is crucial to develop applications that can run on the new multi-core architectures and find out distributions to maximize the potential use of the system. Today most of commercial electronic devices, available in the market, are composed of embedded systems. These devices incorporate recently multi-core processors. Task management onto multiple core/processors is not a trivial issue, and a good task/actor scheduling can yield to significant improvements in terms of efficiency gains and also processor power consumption. Scheduling of data flows between the actors that implement the applications aims to harness multi-core architectures to more types of applications, with an explicit expression of parallelism into the application. On the other hand, the recent development of the MPEG Reconfigurable Video Coding (RVC) standard allows the reconfiguration of the video decoders. RVC is a flexible standard compatible with MPEG developed codecs, making it the ideal tool to integrate into the new multimedia terminals to decode video sequences. With the new versions of the Open RVC-CAL Compiler (Orcc), a static mapping of the actors that implement the functionality of the application can be done once the application executable has been generated. This static mapping must be done for each of the different cores available on the working platform. It has been chosen an embedded system with a processor with two ARMv7 cores. This platform allows us to obtain the desired tests, get as much improvement results from the execution on a single core, and contrast both with a PC-based multiprocessor system. Las posibilidades ofrecidas por el aumento de la velocidad de la frecuencia de reloj de sistemas de un solo procesador están siendo agotadas. Las nuevas arquitecturas multiprocesador proporcionan una vía de desarrollo alternativa en este sentido. El diseño y optimización de aplicaciones de descodificación de video que se ejecuten sobre las nuevas arquitecturas permiten un mejor aprovechamiento y favorecen la obtención de mayores rendimientos. Hoy en día muchos de los dispositivos comerciales que se están lanzando al mercado están integrados por sistemas embebidos, que recientemente están basados en arquitecturas multinúcleo. El manejo de las tareas de ejecución sobre este tipo de arquitecturas no es una tarea trivial, y una buena planificación de los actores que implementan las funcionalidades puede proporcionar importantes mejoras en términos de eficiencia en el uso de la capacidad de los procesadores y, por ende, del consumo de energía. Por otro lado, el reciente desarrollo del estándar de Codificación de Video Reconfigurable (RVC), permite la reconfiguración de los descodificadores de video. RVC es un estándar flexible y compatible con anteriores codecs desarrollados por MPEG. Esto hace de RVC el estándar ideal para ser incorporado en los nuevos terminales multimedia que se están comercializando. Con el desarrollo de las nuevas versiones del compilador específico para el desarrollo de lenguaje RVC-CAL (Orcc), en el que se basa MPEG RVC, el mapeo estático, para entornos basados en multiprocesador, de los actores que integran un descodificador es posible. Se ha elegido un sistema embebido con un procesador con dos núcleos ARMv7. Esta plataforma nos permitirá llevar a cabo las pruebas de verificación y contraste de los conceptos estudiados en este trabajo, en el sentido del desarrollo de descodificadores de video basados en MPEG RVC y del estudio de la planificación y mapeo estático de los mismos.
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The objective of this thesis is model some processes from the nature as evolution and co-evolution, and proposing some techniques that can ensure that these learning process really happens and useful to solve some complex problems as Go game. The Go game is ancient and very complex game with simple rules which still is a challenge for the Artificial Intelligence. This dissertation cover some approaches that were applied to solve this problem, proposing solve this problem using competitive and cooperative co-evolutionary learning methods and other techniques proposed by the author. To study, implement and prove these methods were used some neural networks structures, a framework free available and coded many programs. The techniques proposed were coded by the author, performed many experiments to find the best configuration to ensure that co-evolution is progressing and discussed the results. Using co-evolutionary learning processes can be observed some pathologies which could impact co-evolution progress. In this dissertation is introduced some techniques to solve pathologies as loss of gradients, cycling dynamics and forgetting. According to some authors, one solution to solve these co-evolution pathologies is introduce more diversity in populations that are evolving. In this thesis is proposed some techniques to introduce more diversity and some diversity measurements for neural networks structures to monitor diversity during co-evolution. The genotype diversity evolved were analyzed in terms of its impact to global fitness of the strategies evolved and their generalization. Additionally, it was introduced a memory mechanism in the network neural structures to reinforce some strategies in the genes of the neurons evolved with the intention that some good strategies learned are not forgotten. In this dissertation is presented some works from other authors in which cooperative and competitive co-evolution has been applied. The Go board size used in this thesis was 9x9, but can be easily escalated to more bigger boards.The author believe that programs coded and techniques introduced in this dissertation can be used for other domains.
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This paper analyzes the relationship among research collaboration, number of documents and number of citations of computer science research activity. It analyzes the number of documents and citations and how they vary by number of authors. They are also analyzed (according to author set cardinality) under different circumstances, that is, when documents are written in different types of collaboration, when documents are published in different document types, when documents are published in different computer science subdisciplines, and, finally, when documents are published by journals with different impact factor quartiles. To investigate the above relationships, this paper analyzes the publications listed in the Web of Science and produced by active Spanish university professors between 2000 and 2009, working in the computer science field. Analyzing all documents, we show that the highest percentage of documents are published by three authors, whereas single-authored documents account for the lowest percentage. By number of citations, there is no positive association between the author cardinality and citation impact. Statistical tests show that documents written by two authors receive more citations per document and year than documents published by more authors. In contrast, results do not show statistically significant differences between documents published by two authors and one author. The research findings suggest that international collaboration results on average in publications with higher citation rates than national and institutional collaborations. We also find differences regarding citation rates between journals and conferences, across different computer science subdisciplines and journal quartiles as expected. Finally, our impression is that the collaborative level (number of authors per document) will increase in the coming years, and documents published by three or four authors will be the trend in computer science literature.
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There are a number of research and development activities that are exploring Time and Space Partition (TSP) to implement safe and secure flight software. This approach allows to execute different real-time applications with different levels of criticality in the same computer board. In order to do that, flight applications must be isolated from each other in the temporal and spatial domains. This paper presents the first results of a partitioning platform based on the Open Ravenscar Kernel (ORK+) and the XtratuM hypervisor. ORK+ is a small, reliable real-time kernel supporting the Ada Ravenscar Computational model that is central to the ASSERT development process. XtratuM supports multiple virtual machines, i.e. partitions, on a single computer and is being used in the Integrated Modular Avionics for Space study. ORK+ executes in an XtratuM partition enabling Ada applications to share the computer board with other applications.
Resumo:
Hoy en día el uso de dispositivos portátiles multimedia es ya una realidad totalmente habitual. Además, estos dispositivos tienen una capacidad de cálculo y unos recursos gráficos y de memoria altos, tanto es así que por ejemplo en un móvil se pueden reproducir vídeos de muy alta calidad o tener capacidad para manejar entornos 3D. El precio del uso de estos recursos es un mayor consumo de batería que en ocasiones es demasiado alto y acortan en gran medida la vida de la carga útil de la batería. El Grupo de Diseño Electrónico y Microelectrónico de la Universidad Politécnica de Madrid ha abierto una línea de trabajo que busca la optimización del consumo de energía en este tipo de dispositivos, concretamente en el ámbito de la reproducción de vídeo. El enfoque para afrontar la solución del problema se basa en obtener un mayor rendimiento de la batería a costa de disminuir la experiencia multimedia del usuario. De esta manera, cuando la carga de la batería esté por debajo de un determinado umbral mientras el dispositivo esté reproduciendo un vídeo de alta calidad será el dispositivo quien se autoconfigure dinámicamente para consumir menos potencia en esta tarea, reduciendo la tasa de imágenes por segundo o la resolución del vídeo que se descodifica. Además de lo citado anteriormente se propone dividir la descodificación y la representación del vídeo en dos procesadores, uno de propósito general y otro para procesado digital de señal, con esto se consigue que tener la misma capacidad de cálculo que con un solo procesador pero a una frecuencia menor. Para materializar la propuesta se usará la tarjeta BeagleBoard basada en un procesador multinúcleo OMAP3530 de Texas Instrument que contiene dos núcleos: un ARM1 Cortex-A8 y un DSP2 de la familia C6000. Este procesador multinúcleo además permite modificar la frecuencia de reloj y la tensión de alimentación dinámicamente para conseguir reducir de este modo el consumo del terminal. Por otro lado, como reproductor de vídeos se utilizará una versión de MPlayer que integra un descodificador de vídeo escalable que permite elegir dinámicamente la resolución o las imágenes por segundo que se decodifican para posteriormente mostrarlas. Este reproductor se ejecutará en el núcleo ARM pero debido a la alta carga computacional de la descodificación de vídeos, y que el ARM no está optimizado para este tipo de procesado de datos, el reproductor debe encargar la tarea de la descodificación al DSP. El objetivo de este Proyecto Fin de Carrera consiste en que mientras el descodificador de vídeo está ejecutándose en el núcleo DSP y el Mplayer en el núcleo ARM del OMAP3530 se pueda elegir dinámicamente qué parte del vídeo se descodifica, es decir, seleccionar en tiempo real la calidad o capa del vídeo que se quiere mostrar. Haciendo esto, se podrá quitar carga computacional al núcleo ARM y asignársela al DSP el cuál puede procesarla a menor frecuencia para ahorrar batería. 1 ARM: Es una arquitectura de procesadores de propósito general basada en RISC (Reduced Instruction Set Computer). Es desarrollada por la empresa inglesa ARM holdings. 2 DSP: Procesador Digital de Señal (Digital Signal Processor). Es un sistema basado en procesador, el cual está orientado al cálculo matemático a altas velocidad. Generalmente poseen varias unidades aritmético-lógicas (ALUs) para conseguir realizar varias operaciones simultáneamente. SUMMARY. Nowadays, the use of multimedia devices is a well known reality. In addition, these devices have high graphics and calculus performance and a lot of memory as well. In instance, we can play high quality videos and 3D environments in a mobile phone. That kind of use may increase the device's power consumption and make shorter the battery duration. Electronic and Microelectronic Design Group of Technical University of Madrid has a research line which is looking for optimization of power consumption while these devices are playing videos. The solution of this trouble is based on taking more advantage of battery by decreasing multimedia user experience. On this way, when battery charge is under a threshold while device is playing a high quality video the device is going to configure itself dynamically in order to decrease its power consumption by decreasing frame per second rate, video resolution or increasing the noise in the decoded frame. It is proposed splitting decoding and representation tasks in two processors in order to have the same calculus capability with lower frecuency. The first one is specialized in digital signal processing and the other one is a general purpose processor. In order to materialize this proposal we will use a board called BeagleBoard which is based on a multicore processor called OMAP3530 from Texas Instrument. This processor includes two cores: ARM Cortex-A8 and a TMS320C64+ DSP core. Changing clock frequency and supply voltage is allowed by OMAP3530, we can decrease the power consumption on this way. On the other hand, MPlayer will be used as video player. It includes a scalable video decoder which let us changing dynamically the resolution or frames per second rate of the video in order to show it later. This player will be executed by ARM core but this is not optimized for this task, for that reason, DSP core will be used to decoding video. The target of this final career project is being able to choose which part of the video is decoded each moment while decoder is executed by DSP and Mplayer by ARM. It will be able to change in real time the video quality, resolution and frames per second that user want to show. On this way, reducing the computational charge within the processor will be possible.
Resumo:
An inverse optimization strategy was developed to determine the single crystal properties from experimental results of the mechanical behavior of polycrystals. The polycrystal behavior was obtained by means of the finite element simulation of a representative volume element of the microstructure in which the dominant slip and twinning systems were included in the constitutive equation of each grain. The inverse problem was solved by means of the Levenberg-Marquardt method, which provided an excellent fit to the experimental results. The iterative optimization process followed a hierarchical scheme in which simple representative volume elements were initially used, followed by more realistic ones to reach the final optimum solution, leading to important reductions in computer time. The new strategy was applied to identify the initial and saturation critical resolved shear stresses and the hardening modulus of the active slip systems and extension twinning in a textured AZ31 Mg alloy. The results were in general agreement with the data in the literature but also showed some differences. They were partially explained because of the higher accuracy of the new optimization strategy but it was also shown that the number of independent experimental stress-strain curves used as input is critical to reach an accurate solution to the inverse optimization problem. It was concluded that at least three independent stress-strain curves are necessary to determine the single crystal behavior from polycrystal tests in the case of highly textured Mg alloys.
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Este proyecto consiste en el diseño e implementación un sistema domótico que puede ser instalado en una vivienda para controlar distintas variables ambientales y conseguir así la máxima comodidad de los habitantes de manera automática o manual según los gustos y necesidades de los usuarios. La característica principal de este sistema, es que cuenta con un funcionamiento distribuido donde entran en juego un servidor, encargado de tomar las decisiones generales para el comportamiento de la casa, y una serie de controladores esclavo cuya función es mantener constantes las variables ambientales con los valores fijados por el servidor. Así se consigue mantener la vivienda en una situación de bienestar constante para cualquier persona que se encuentre dentro. El sistema ha sido pensado de manera que se intenta reducir al máximo el cableado para facilitar su instalación por lo que la comunicación entre los distintos dispositivos se hace de manera inalámbrica por medio de un protocolo descrito en la norma IEEE 802.15.4 llamado ZigBee. Para ello se ha utilizado un módulo de comunicación wireless llamado Xbee, el cual permite la comunicación entre dos dispositivos. Para el control de dicho sistema distribuido se cuenta con una aplicación web, que mediante una interfaz gráfica permite al usuario controlar los distintos dispositivos dentro de la vivienda consiguiendo así controlar las variables ambientales a gusto del usuario. Dicha interfaz gráfica no depende de un software específico, sino que sólo es necesario un cliente http como podría ser Internet Explorer, Mozilla Firefox, Google Chrome, etc. Para integrar dicho sistema se ha usado un mini ordenador de bajo coste llamado RaspBerryPi, en el que se encuentra alojado un servidor Apache con el fin de gestionar y automatizar las variables ambientales. El control de los dipositivos encargados de modificar y estabilizar las variables ambientales se realiza mediante unos controladores genéricos implementados mediante mcontroladores 80C51F410, pertenecientes a la familia 80C51, y una serie de componentes y circuitería que permiten el correcto funcionamiento de éstos. Existen dos tipos de controladores distintos, los cuales son: Controlador Sensor: Encargados de las tomas de valores ambientales como puede ser la luz y la temperatura. Controladores Actuadores: Encargados de actuar sobre los dispositivos que modifican y estabilizan las variables ambientales como pueden ser la calefacción, tiras de leds de iluminación, persianas, alarmas, etc. El conjunto de la RaspBerryPi y los diferentes controladores forman el prototipo diseñado para este proyecto fin de carrera, el cual puede ser ampliado sencillamente para abarcar una amplia gama de posibilidades y funcionalidades dentro de la comodidad de una vivienda. ABSTRACT. The project described in this report consisted designing and implementing a home automation system that could be installed in a house in order to control environmental variables and thus get the maximum comfort of the inhabitant automatically or manually according to their tastes and needs. The main feature of this system consists in a distributed system, formed by a server which is responsible for making the main decisions of the actions performed inside the house. In addition, there are a series of slave controlers whose function consists in keeping the environmental variables within the values established by the server. Thus gets to keep the home in a situation of constant wellbeing to anyone who is inside. The system has been designed in order to reduce the amount of wire needed for the inter-connection of the devices, by means of wireless communication. The devices chosen for the solution are Xbee modules, which use the Zigbee protocol in order to comunicate one between each other. The Zigbee protocol is fully described in the IEEE 802.15.4 standard. A web application has been used to control the distributed system. This application allows users to control various devices inside the house and subsequently the different environmental variables. This implementation allows obtaining the maximum comfort by means of a very simple graphical interface. In addition, the Graphical User Interface (GUI) does not depend on any specific software. This means that it would only be necessary a http client (such as Internet Explorer, Mozilla Firefox, Google Chrome, etc.) for handling the application. The system has been integrated using a low-cost mini computer called RaspBerryPi.This computer has an Apache server allocated which allows to manage and to automatize the different environmental variables. Furthermore, for changing and stabilizing those variables, some generic controllers have been developed, based on mcontrollers 80C51F410. There have been developed mainly two different types of controllers: Sensor Controllers, responsible for measuring the different environmental values, such as light and temperature; and Actuator Controllers, which purpose is to modify and stabilize those environmental variables by actuating on the heating, the led lamps, the blinders, the alarm, etc. The combination of the RaspBerryPi and the different controllers conform the prototype designed during this project. Additionally, this solution could be easily expanded in order to intake further functionalities adapted to new needs that could arise in the future.
Resumo:
In this paper we tackle the problem of landing a helicopter autonomously on a ship deck, using as the main sensor, an on-board colour camera. To create a test-bed, we first adequately simulate the movement of a ship landing platform on the Sea, for different Sea States, for different ships, randomly and realistically enough. We use a commercial parallel robot to get this movement. Once we had this, we developed an accurate and robust computer vision system to measure the pose of the helipad with respect to the on-board camera. To deal with the noise and the possible fails of the computer vision, a state estimator was created. With all of this, we are now able to develop and test a controller that closes the loop and finish the autonomous landing task.
Resumo:
La evolución de los teléfonos móviles inteligentes, dotados de cámaras digitales, está provocando una creciente demanda de aplicaciones cada vez más complejas que necesitan algoritmos de visión artificial en tiempo real; puesto que el tamaño de las señales de vídeo no hace sino aumentar y en cambio el rendimiento de los procesadores de un solo núcleo se ha estancado, los nuevos algoritmos que se diseñen para visión artificial han de ser paralelos para poder ejecutarse en múltiples procesadores y ser computacionalmente escalables. Una de las clases de procesadores más interesantes en la actualidad se encuentra en las tarjetas gráficas (GPU), que son dispositivos que ofrecen un alto grado de paralelismo, un excelente rendimiento numérico y una creciente versatilidad, lo que los hace interesantes para llevar a cabo computación científica. En esta tesis se exploran dos aplicaciones de visión artificial que revisten una gran complejidad computacional y no pueden ser ejecutadas en tiempo real empleando procesadores tradicionales. En cambio, como se demuestra en esta tesis, la paralelización de las distintas subtareas y su implementación sobre una GPU arrojan los resultados deseados de ejecución con tasas de refresco interactivas. Asimismo, se propone una técnica para la evaluación rápida de funciones de complejidad arbitraria especialmente indicada para su uso en una GPU. En primer lugar se estudia la aplicación de técnicas de síntesis de imágenes virtuales a partir de únicamente dos cámaras lejanas y no paralelas—en contraste con la configuración habitual en TV 3D de cámaras cercanas y paralelas—con información de color y profundidad. Empleando filtros de mediana modificados para la elaboración de un mapa de profundidad virtual y proyecciones inversas, se comprueba que estas técnicas son adecuadas para una libre elección del punto de vista. Además, se demuestra que la codificación de la información de profundidad con respecto a un sistema de referencia global es sumamente perjudicial y debería ser evitada. Por otro lado se propone un sistema de detección de objetos móviles basado en técnicas de estimación de densidad con funciones locales. Este tipo de técnicas es muy adecuada para el modelado de escenas complejas con fondos multimodales, pero ha recibido poco uso debido a su gran complejidad computacional. El sistema propuesto, implementado en tiempo real sobre una GPU, incluye propuestas para la estimación dinámica de los anchos de banda de las funciones locales, actualización selectiva del modelo de fondo, actualización de la posición de las muestras de referencia del modelo de primer plano empleando un filtro de partículas multirregión y selección automática de regiones de interés para reducir el coste computacional. Los resultados, evaluados sobre diversas bases de datos y comparados con otros algoritmos del estado del arte, demuestran la gran versatilidad y calidad de la propuesta. Finalmente se propone un método para la aproximación de funciones arbitrarias empleando funciones continuas lineales a tramos, especialmente indicada para su implementación en una GPU mediante el uso de las unidades de filtraje de texturas, normalmente no utilizadas para cómputo numérico. La propuesta incluye un riguroso análisis matemático del error cometido en la aproximación en función del número de muestras empleadas, así como un método para la obtención de una partición cuasióptima del dominio de la función para minimizar el error. ABSTRACT The evolution of smartphones, all equipped with digital cameras, is driving a growing demand for ever more complex applications that need to rely on real-time computer vision algorithms. However, video signals are only increasing in size, whereas the performance of single-core processors has somewhat stagnated in the past few years. Consequently, new computer vision algorithms will need to be parallel to run on multiple processors and be computationally scalable. One of the most promising classes of processors nowadays can be found in graphics processing units (GPU). These are devices offering a high parallelism degree, excellent numerical performance and increasing versatility, which makes them interesting to run scientific computations. In this thesis, we explore two computer vision applications with a high computational complexity that precludes them from running in real time on traditional uniprocessors. However, we show that by parallelizing subtasks and implementing them on a GPU, both applications attain their goals of running at interactive frame rates. In addition, we propose a technique for fast evaluation of arbitrarily complex functions, specially designed for GPU implementation. First, we explore the application of depth-image–based rendering techniques to the unusual configuration of two convergent, wide baseline cameras, in contrast to the usual configuration used in 3D TV, which are narrow baseline, parallel cameras. By using a backward mapping approach with a depth inpainting scheme based on median filters, we show that these techniques are adequate for free viewpoint video applications. In addition, we show that referring depth information to a global reference system is ill-advised and should be avoided. Then, we propose a background subtraction system based on kernel density estimation techniques. These techniques are very adequate for modelling complex scenes featuring multimodal backgrounds, but have not been so popular due to their huge computational and memory complexity. The proposed system, implemented in real time on a GPU, features novel proposals for dynamic kernel bandwidth estimation for the background model, selective update of the background model, update of the position of reference samples of the foreground model using a multi-region particle filter, and automatic selection of regions of interest to reduce computational cost. The results, evaluated on several databases and compared to other state-of-the-art algorithms, demonstrate the high quality and versatility of our proposal. Finally, we propose a general method for the approximation of arbitrarily complex functions using continuous piecewise linear functions, specially formulated for GPU implementation by leveraging their texture filtering units, normally unused for numerical computation. Our proposal features a rigorous mathematical analysis of the approximation error in function of the number of samples, as well as a method to obtain a suboptimal partition of the domain of the function to minimize approximation error.
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In the major pathway of homologous DNA recombination in prokaryotic cells, the Holliday junction intermediate is processed through its association with RuvA, RuvB, and RuvC proteins. Specific binding of the RuvA tetramer to the Holliday junction is required for the RuvB motor protein to be loaded onto the junction DNA, and the RuvAB complex drives the ATP-dependent branch migration. We solved the crystal structure of the Holliday junction bound to a single Escherichia coli RuvA tetramer at 3.1-Å resolution. In this complex, one side of DNA is accessible for cleavage by RuvC resolvase at the junction center. The refined junction DNA structure revealed an open concave architecture with a four-fold symmetry. Each arm, with B-form DNA, in the Holliday junction is predominantly recognized in the minor groove through hydrogen bonds with two repeated helix-hairpin-helix motifs of each RuvA subunit. The local conformation near the crossover point, where two base pairs are disrupted, suggests a possible scheme for successive base pair rearrangements, which may account for smooth Holliday junction movement without segmental unwinding.
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Random walks have been used to describe a wide variety of systems ranging from cell colonies to polymers. Sixty-five years ago, Kuhn [Kuhn, W. (1934) Kolloid-Z. 68, 2–11] made the prediction, backed later by computer simulations, that the overall shape of a random-walk polymer is aspherical, yet no experimental work has directly tested Kuhn's general idea and subsequent computer simulations. By using fluorescence microscopy, we monitored the conformation of individual, long, random-walk polymers (fluorescently labeled DNA molecules) at equilibrium. We found that a polymer most frequently adopts highly extended, nonfractal structures with a strongly anisotropic shape. The ensemble-average ratio of the lengths of the long and short axes of the best-fit ellipse of the polymer was much larger than unity.
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Computer models were used to examine whether and under what conditions the multimeric protein complex is inhibited by high concentrations of one of its components—an effect analogous to the prozone phenomenon in precipitin tests. A series of idealized simple “ball-and-stick” structures representing small oligomeric complexes of protein molecules formed by reversible binding reactions were analyzed to determine the binding steps leading to each structure. The equilibrium state of each system was then determined over a range of starting concentrations and Kds and the steady-state concentration of structurally complete oligomer calculated for each situation. A strong inhibitory effect at high concentrations was shown by any protein molecule forming a bridge between two or more separable parts of the complex. By contrast, proteins linked to the outside of the complex by a single bond showed no inhibition whatsoever at any concentration. Nonbridging, multivalent proteins in the body of the complex could show an inhibitory effect or not depending on the structure of the complex and the strength of its bonds. On the basis of this study, we suggest that the prozone phenomenon will occur widely in living cells and that it could be a crucial factor in the regulation of protein complex formation.
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5′-End fragments of two genes encoding plastid-localized acetyl-CoA carboxylase (ACCase; EC 6.4.1.2) of wheat (Triticum aestivum) were cloned and sequenced. The sequences of the two genes, Acc-1,1 and Acc-1,2, are 89% identical. Their exon sequences are 98% identical. The amino acid sequence of the biotin carboxylase domain encoded by Acc-1,1 and Acc-1,2 is 93% identical with the maize plastid ACCase but only 80–84% identical with the cytosolic ACCases from other plants and from wheat. Four overlapping fragments of cDNA covering the entire coding region were cloned by PCR and sequenced. The wheat plastid ACCase ORF contains 2,311 amino acids with a predicted molecular mass of 255 kDa. A putative transit peptide is present at the N terminus. Comparison of the genomic and cDNA sequences revealed introns at conserved sites found in the genes of other plant multifunctional ACCases, including two introns absent from the wheat cytosolic ACCase genes. Transcription start sites of the plastid ACCase genes were estimated from the longest cDNA clones obtained by 5′-RACE (rapid amplification of cDNA ends). The untranslated leader sequence encoded by the Acc-1 genes is at least 130–170 nucleotides long and is interrupted by an intron. Southern analysis indicates the presence of only one copy of the gene in each ancestral chromosome set. The gene maps near the telomere on the short arm of chromosomes 2A, 2B, and 2D. Identification of three different cDNAs, two corresponding to genes Acc-1,1 and Acc-1,2, indicates that all three genes are transcriptionally active.