A new feature extraction method for signal classification applied to cat spinal cord signals

In the spinal cord of the anesthetized cat, spontaneous cord dorsum potentials (CDPs) appear synchronously along the lumbo-sacral segments. These CDPs have different shapes and magnitudes. Previous work has indicated that some CDPs appear to be specially associated with the activation of spinal pathways that lead to primary afferent depolarization and presynaptic inhibition. Visual detection and classification of these CDPs provides relevant information on the functional organization of the neural networks involved in the control of sensory information and allows the characterization of the changes produced by acute nerve and spinal lesions. We now present a novel feature extraction approach for signal classification, applied to CDP detection. The method is based on an intuitive procedure. We first remove by convolution the noise from the CDPs recorded in each given spinal segment. Then, we assign a coefficient for each main local maximum of the signal using its amplitude and distance to the most important maximum of the signal. These coefficients will be the input for the subsequent classification algorithm. In particular, we employ gradient boosting classification trees. This combination of approaches allows a faster and more accurate discrimination of CDPs than is obtained by other methods.

Design and control of multi-finger haptic devices for dexterous manipulation

En la interacción con el entorno que nos rodea durante nuestra vida diaria (utilizar un cepillo de dientes, abrir puertas, utilizar el teléfono móvil, etc.) y en situaciones profesionales (intervenciones médicas, procesos de producción, etc.), típicamente realizamos manipulaciones avanzadas que incluyen la utilización de los dedos de ambas manos. De esta forma el desarrollo de métodos de interacción háptica multi-dedo dan lugar a interfaces hombre-máquina más naturales y realistas. No obstante, la mayoría de interfaces hápticas disponibles en el mercado están basadas en interacciones con un solo punto de contacto; esto puede ser suficiente para la exploración o palpación del entorno pero no permite la realización de tareas más avanzadas como agarres. En esta tesis, se investiga el diseño mecánico, control y aplicaciones de dispositivos hápticos modulares con capacidad de reflexión de fuerzas en los dedos índice, corazón y pulgar del usuario. El diseño mecánico de la interfaz diseñada, ha sido optimizado con funciones multi-objetivo para conseguir una baja inercia, un amplio espacio de trabajo, alta manipulabilidad y reflexión de fuerzas superiores a 3 N en el espacio de trabajo. El ancho de banda y la rigidez del dispositivo se han evaluado mediante simulación y experimentación real. Una de las áreas más importantes en el diseño de estos dispositivos es el efector final, ya que es la parte que está en contacto con el usuario. Durante este trabajo se ha diseñado un dedal de bajo peso, adaptable a diferentes usuarios que, mediante la incorporación de sensores de contacto, permite estimar fuerzas normales y tangenciales durante la interacción con entornos reales y virtuales. Para el diseño de la arquitectura de control, se estudiaron los principales requisitos para estos dispositivos. Entre estos, cabe destacar la adquisición, procesado e intercambio a través de internet de numerosas señales de control e instrumentación; la computación de equaciones matemáticas incluyendo la cinemática directa e inversa, jacobiana, algoritmos de detección de agarres, etc. Todos estos componentes deben calcularse en tiempo real garantizando una frecuencia mínima de 1 KHz. Además, se describen sistemas para manipulación de precisión virtual y remota; así como el diseño de un método denominado "desacoplo cinemático iterativo" para computar la cinemática inversa de robots y la comparación con otros métodos actuales. Para entender la importancia de la interacción multimodal, se ha llevado a cabo un estudio para comprobar qué estímulos sensoriales se correlacionan con tiempos de respuesta más rápidos y de mayor precisión. Estos experimentos se desarrollaron en colaboración con neurocientíficos del instituto Technion Israel Institute of Technology. Comparando los tiempos de respuesta en la interacción unimodal (auditiva, visual y háptica) con combinaciones bimodales y trimodales de los mismos, se demuestra que el movimiento sincronizado de los dedos para generar respuestas de agarre se basa principalmente en la percepción háptica. La ventaja en el tiempo de procesamiento de los estímulos hápticos, sugiere que los entornos virtuales que incluyen esta componente sensorial generan mejores contingencias motoras y mejoran la credibilidad de los eventos. Se concluye que, los sistemas que incluyen percepción háptica dotan a los usuarios de más tiempo en las etapas cognitivas para rellenar información de forma creativa y formar una experiencia más rica. Una aplicación interesante de los dispositivos hápticos es el diseño de nuevos simuladores que permitan entrenar habilidades manuales en el sector médico. En colaboración con fisioterapeutas de Griffith University en Australia, se desarrolló un simulador que permite realizar ejercicios de rehabilitación de la mano. Las propiedades de rigidez no lineales de la articulación metacarpofalange del dedo índice se estimaron mediante la utilización del efector final diseñado. Estos parámetros, se han implementado en un escenario que simula el comportamiento de la mano humana y que permite la interacción háptica a través de esta interfaz. Las aplicaciones potenciales de este simulador están relacionadas con entrenamiento y educación de estudiantes de fisioterapia. En esta tesis, se han desarrollado nuevos métodos que permiten el control simultáneo de robots y manos robóticas en la interacción con entornos reales. El espacio de trabajo alcanzable por el dispositivo háptico, se extiende mediante el cambio de modo de control automático entre posición y velocidad. Además, estos métodos permiten reconocer el gesto del usuario durante las primeras etapas de aproximación al objeto para su agarre. Mediante experimentos de manipulación avanzada de objetos con un manipulador y diferentes manos robóticas, se muestra que el tiempo en realizar una tarea se reduce y que el sistema permite la realización de la tarea con precisión. Este trabajo, es el resultado de una colaboración con investigadores de Harvard BioRobotics Laboratory. ABSTRACT When we interact with the environment in our daily life (using a toothbrush, opening doors, using cell-phones, etc.), or in professional situations (medical interventions, manufacturing processes, etc.) we typically perform dexterous manipulations that involve multiple fingers and palm for both hands. Therefore, multi-Finger haptic methods can provide a realistic and natural human-machine interface to enhance immersion when interacting with simulated or remote environments. Most commercial devices allow haptic interaction with only one contact point, which may be sufficient for some exploration or palpation tasks but are not enough to perform advanced object manipulations such as grasping. In this thesis, I investigate the mechanical design, control and applications of a modular haptic device that can provide force feedback to the index, thumb and middle fingers of the user. The designed mechanical device is optimized with a multi-objective design function to achieve a low inertia, a large workspace, manipulability, and force-feedback of up to 3 N within the workspace; the bandwidth and rigidity for the device is assessed through simulation and real experimentation. One of the most important areas when designing haptic devices is the end-effector, since it is in contact with the user. In this thesis the design and evaluation of a thimble-like, lightweight, user-adaptable, and cost-effective device that incorporates four contact force sensors is described. This design allows estimation of the forces applied by a user during manipulation of virtual and real objects. The design of a real-time, modular control architecture for multi-finger haptic interaction is described. Requirements for control of multi-finger haptic devices are explored. Moreover, a large number of signals have to be acquired, processed, sent over the network and mathematical computations such as device direct and inverse kinematics, jacobian, grasp detection algorithms, etc. have to be calculated in Real Time to assure the required high fidelity for the haptic interaction. The Hardware control architecture has different modules and consists of an FPGA for the low-level controller and a RT controller for managing all the complex calculations (jacobian, kinematics, etc.); this provides a compact and scalable solution for the required high computation capabilities assuring a correct frequency rate for the control loop of 1 kHz. A set-up for dexterous virtual and real manipulation is described. Moreover, a new algorithm named the iterative kinematic decoupling method was implemented to solve the inverse kinematics of a robotic manipulator. In order to understand the importance of multi-modal interaction including haptics, a subject study was carried out to look for sensory stimuli that correlate with fast response time and enhanced accuracy. This experiment was carried out in collaboration with neuro-scientists from Technion Israel Institute of Technology. By comparing the grasping response times in unimodal (auditory, visual, and haptic) events with the response times in events with bimodal and trimodal combinations. It is concluded that in grasping tasks the synchronized motion of the fingers to generate the grasping response relies on haptic cues. This processing-speed advantage of haptic cues suggests that multimodalhaptic virtual environments are superior in generating motor contingencies, enhancing the plausibility of events. Applications that include haptics provide users with more time at the cognitive stages to fill in missing information creatively and form a richer experience. A major application of haptic devices is the design of new simulators to train manual skills for the medical sector. In collaboration with physical therapists from Griffith University in Australia, we developed a simulator to allow hand rehabilitation manipulations. First, the non-linear stiffness properties of the metacarpophalangeal joint of the index finger were estimated by using the designed end-effector; these parameters are implemented in a scenario that simulates the behavior of the human hand and that allows haptic interaction through the designed haptic device. The potential application of this work is related to educational and medical training purposes. In this thesis, new methods to simultaneously control the position and orientation of a robotic manipulator and the grasp of a robotic hand when interacting with large real environments are studied. The reachable workspace is extended by automatically switching between rate and position control modes. Moreover, the human hand gesture is recognized by reading the relative movements of the index, thumb and middle fingers of the user during the early stages of the approximation-to-the-object phase and then mapped to the robotic hand actuators. These methods are validated to perform dexterous manipulation of objects with a robotic manipulator, and different robotic hands. This work is the result of a research collaboration with researchers from the Harvard BioRobotics Laboratory. The developed experiments show that the overall task time is reduced and that the developed methods allow for full dexterity and correct completion of dexterous manipulations.

Optically programmable logic cells as basic units for transmission and synchronization of chaotic signals

We proposed an optical communications system, based on a digital chaotic signal where the synchronization of chaos was the main objective, in some previous papers. In this paper we will extend this work. A way to add the digital data signal to be transmitted onto the chaotic signal and its correct reception, is the main objective. We report some methods to study the main characteristics of the resulting signal. The main problem with any real system is the presence of some retard between the times than the signal is generated at the emitter at the time when this signal is received. Any system using chaotic signals as a method to encrypt need to have the same characteristics in emitter and receiver. It is because that, this control of time is needed. A method to control, in real time the chaotic signals, is reported.

Variable impedance LED matrix control system for lamps in PV applications

The interest in LED lighting has been growing recently due to the high efficacy, lifelime and ruggedness that this technology offers. However the key element to guarantee those parameters with these new electronic devices is to keep under control the working temperature of the semiconductor crystal. This paper propases a LED lamp design that fulfils the requ irements of a PV lighting systems, whose main quality criteria is reliability. It uses directly as a power supply a non·stabilized constant voltage source, as batteries. An electronic control architecture is used to regulate the current applied to the LEO matri)( according to their temperature and the voltage output value of the batteries with two pulse modulation signals (PWM) signals. The first one connects and disconnects the LEOs to the power supply and the second one connects and disconnects several emitters to the electric circuit changing its overall impedance. A prototype of the LEO lamp has been implemented and tested at different temperaturas and battery voltages.

Modelling and Control of Stepper Motors for High Accuracy Positioning Systems Used in Radioactive Environments

Hybrid Stepper Motors are widely used in open-loop position applications. They are the choice of actuation for the collimators in the Large Hadron Collider, the largest particle accelerator at CERN. In this case the positioning requirements and the highly radioactive operating environment are unique. The latter forces both the use of long cables to connect the motors to the drives which act as transmission lines and also prevents the use of standard position sensors. However, reliable and precise operation of the collimators is critical for the machine, requiring the prevention of step loss in the motors and maintenance to be foreseen in case of mechanical degradation. In order to make the above possible, an approach is proposed for the application of an Extended Kalman Filter to a sensorless stepper motor drive, when the motor is separated from its drive by long cables. When the long cables and high frequency pulse width modulated control voltage signals are used together, the electrical signals difer greatly between the motor and drive-side of the cable. Since in the considered case only drive-side data is available, it is therefore necessary to estimate the motor-side signals. Modelling the entire cable and motor system in an Extended Kalman Filter is too computationally intensive for standard embedded real-time platforms. It is, in consequence, proposed to divide the problem into an Extended Kalman Filter, based only on the motor model, and separated motor-side signal estimators, the combination of which is less demanding computationally. The efectiveness of this approach is shown in simulation. Then its validity is experimentally demonstrated via implementation in a DSP based drive. A testbench to test its performance when driving an axis of a Large Hadron Collider collimator is presented along with the results achieved. It is shown that the proposed method is capable of achieving position and load torque estimates which allow step loss to be detected and mechanical degradation to be evaluated without the need for physical sensors. These estimation algorithms often require a precise model of the motor, but the standard electrical model used for hybrid stepper motors is limited when currents, which are high enough to produce saturation of the magnetic circuit, are present. New model extensions are proposed in order to have a more precise model of the motor independently of the current level, whilst maintaining a low computational cost. It is shown that a significant improvement in the model It is achieved with these extensions, and their computational performance is compared to study the cost of model improvement versus computation cost. The applicability of the proposed model extensions is demonstrated via their use in an Extended Kalman Filter running in real-time for closed-loop current control and mechanical state estimation. An additional problem arises from the use of stepper motors. The mechanics of the collimators can wear due to the abrupt motion and torque profiles that are applied by them when used in the standard way, i.e. stepping in open-loop. Closed-loop position control, more specifically Field Oriented Control, would allow smoother profiles, more respectful to the mechanics, to be applied but requires position feedback. As mentioned already, the use of sensors in radioactive environments is very limited for reliability reasons. Sensorless control is a known option but when the speed is very low or zero, as is the case most of the time for the motors used in the LHC collimator, the loss of observability prevents its use. In order to allow the use of position sensors without reducing the long term reliability of the whole system, the possibility to switch from closed to open loop is proposed and validated, allowing the use of closed-loop control when the position sensors function correctly and open-loop when there is a sensor failure. A different approach to deal with the switched drive working with long cables is also presented. Switched mode stepper motor drives tend to have poor performance or even fail completely when the motor is fed through a long cable due to the high oscillations in the drive-side current. The design of a stepper motor output fillter which solves this problem is thus proposed. A two stage filter, one devoted to dealing with the diferential mode and the other with the common mode, is designed and validated experimentally. With this ?lter the drive performance is greatly improved, achieving a positioning repeatability even better than with the drive working without a long cable, the radiated emissions are reduced and the overvoltages at the motor terminals are eliminated.

Diseño electrónico de la tarjeta de interfaz y control para un equipo de seguimiento en elevación en un subsistema de contramedidas electrónicas

Como apoyo al Subsistema ECM tradicional de los sistemas de Guerra Electrónica, surge el Equipo de Seguimiento en Elevación cuyo fin es la búsqueda de la amenaza en elevación. Ante la necesidad de un mayor número de líneas entre los bloques internos de este Equipo de Seguimiento, se hace necesario el diseño de una Tarjeta de Interfaz y Control. La tarjeta se ocupará de realizar el control de datos y flujo de señales de módulos RF que componen el Equipo, haciendo de interfaz entre las Tarjetas Procesadoras y Digitalizadoras, así como entre la Tarjeta de Proceso y el Bloque de Recepción. Una vez que se haya realizado el análisis de la necesidad de controlar estas señales, estudiado las especificaciones y los requisitos funcionales del Sistema, se procederá a implementar el diseño hardware y desarrollo firmware de la Tarjeta de Interfaz y Control de un Equipo de Seguimiento en Elevación para un Subsistema ECM, que constituye el objetivo de este PFC. ABSTRACT. In order to find a threat on elevation, and as a support on the basic ECM subsystems of E-war systems, the Elevation Tracking Equipment appears. To the need of a bigger number of lines between inside blocks of this Tracking Equipment, the design of an Interface and Control Board is needed. This board will deal with the data control and with the flow of RF modules signals that set up the Equipment, and will also work as an interface between the Processing and Digitalizing Board, as well as between the Process Board and the Reception Block. Once the signal control necessity analysis has been made, and once the specifications and functional System requirements have been studied, the hardware design and the firmware development will be ran as a target of PFC.

Desarrollo de un sistema automático de medida para el control del consumo en sistemas empotrados

El Grupo de Diseño Electrónico y Microelectrónico de la Universidad Politécnica de Madrid -GDEM- se dedica, entre otras cosas, al estudio y mejora del consumo en sistemas empotrados. Es en este lugar y sobre este tema donde el proyecto a exponer ha tomado forma y desarrollo. Según un artículo de la revista online Revista de Electrónica Embebida, un sistema empotrado o embebido es aquel “sistema controlado por un microprocesador y que gracias a la programación que incorpora o que se le debe incorporar, realiza una función específica para la que ha sido diseñado, integrando en su interior la mayoría de los elementos necesarios para realizar dicho función”. El porqué de estudiar sobre este tema responde a que, cada vez, hay mayor presencia de sistemas empotrados en nuestra vida cotidiana. Esto es debido a que se está tendiendo a dotar de “inteligencia” a todo lo que puedan hacer nuestra vida un poco más fácil. Nos podemos encontrar dichos sistemas en fábricas, oficinas de atención a los ciudadanos, sistemas de seguridad de hogar, relojes, móviles, lavadoras, hornos, aspiradores y un largo etcétera en cualquier aparato que nos podamos imaginar. A pesar de sus grandes ventajas, aún hay grandes inconvenientes. El mayor problema que supone a día de hoy es la autonomía del mismo sistema, ya que hablamos de aparatos que muchas veces están alimentados por baterías -para ayudar a su portabilidad–. Por esto, se está intentando dotar a dichos sistemas de una capacidad de ahorro de energía y toma de decisiones que podrían ayudar a duplicar la autonomía de dicha batería. Un ejemplo claro son los Smartphones de hoy en día, unos aparatos casi indispensables que pueden tener una autonomía de un día. Esto es poco práctico para el usuario en caso de viajes, trabajo u otras situaciones en las que se le dé mucho uso y no pueda tener acceso a una red eléctrica. Es por esto que surge la necesidad de investigar, sin necesidad de mejorar el hardware del sistema, una manera de mejorar esta situación. Este proyecto trabajará en esa línea creando un sistema automático de medida el cual generará las corrientes que servirán como entrada para verificar el sistema de adquisición que junto con la tarjeta Beagle Board permitirá la toma de decisiones en relación con el consumo de energía. Para realizar este sistema, nos ayudaremos de diferentes herramientas que podremos encontrar en el laboratorio del GDEM, como la fuente de alimentación Agilent y la Beagle Board –como principales herramientas de trabajo- . El objetivo principal será la simulación de unas señales que, después de pasar un proceso de conversión y tratado, harán la función de representación del consumo de cada una de las partes que pueden formar un sistema empotrado genérico. Por lo tanto, podemos decir que el sistema hará la funcionalidad de un banco de pruebas que ayudará a simular dicho consumo para que el microprocesador del sistema pueda llegar a tomar alguna decisión. ABSTRACT. The Electronic and Microelectronic Design Group of Universidad Politécnica de Madrid -GDEM- is in charge, between other issues, of improving the embedded system’s consumption. It is in this place and about this subject where the exposed project has taken shape and development. According to an article from de online magazine Revista de Electronica Embebida, an embedded system is “the one controlled by a microprocessor and, thanks to the programing that it includes, it carries out a specific function what it has been designed for, being integrated in it the most necessary elements for realizing the already said function”. The because of studying this subject, answers that each time there is more presence of the embedded system in our daily life. This is due to the tendency of providing “intelligence” to all what can make our lives easier. We can find this kind of systems in factories, offices, security systems, watchers, mobile phones, washing machines, ovens, hoovers and, definitely, in all kind of machines what we can think of. Despite its large vantages, there are still some inconveniences. Nowadays, the most important problem is the autonomy of the system itself when machines that have to be supplied by batteries –making easier the portability-. Therefore, this project is going after a save capacity of energy for the system as well as being able to take decisions in order to duplicate batteries’ autonomy. Smartphones are a clear example. They are a very successful product but the autonomy is just one day. This is not practical for users, at all, if they have to travel, to work or to do any activity that involves a huge use of the phone without a socket nearby. That is why the need of investigating a way to improve this situation. This project is working on this line, creating an automatic system that will generate the currents for verifying the acquisition system that, with the beagle board, will help taking decisions regarding the energy’s consumption. To carry out this system, we need different tools that we can find in the laboratory of the group previously mentioned, like power supply Agilent and the Beagle Board – as main working tools –. The main goal is the simulation of some signals that, after a conversion process, will represent de consumption of each of the parts in the embedded generic system. Therefore, the system will be a testing ground that simulate the consumption, once sent to the processor, to be processed and so the microprocessor system might take some decision.

Real-time bilateral control of a nonlinear teleoperation system

In this work, we study the bilateral control of a nonlinear teleoperator system with constant delay, proposes a control strategy by state convergence, which directly connect the local and remote manipulator through feedback signals of position and speed. The control signal allows the remote manipulator follow the local manipulator through the state convergence even if it has a delay in the communication channel. The bilateral control of the teleoperator system considers the case when the human operator applies a constant force on the local manipulator and when the interaction of the remote manipulator with the environment is considered passive. The stability analysis is performed using functional of Lyapunov-Krasovskii, it showed that using a control algorithm by state convergence for the case with constant delay, the nonlinear local and remote teleoperation system is asymptotically stable, also speeds converge to zero and position tracking is achieved.

Automated wordlength optimization framework for multi-source statistical interval-based analysis of nonlinear systems with control-flow structures

El uso de aritmética de punto fijo es una opción de diseño muy extendida en sistemas con fuertes restricciones de área, consumo o rendimiento. Para producir implementaciones donde los costes se minimicen sin impactar negativamente en la precisión de los resultados debemos llevar a cabo una asignación cuidadosa de anchuras de palabra. Encontrar la combinación óptima de anchuras de palabra en coma fija para un sistema dado es un problema combinatorio NP-hard al que los diseñadores dedican entre el 25 y el 50 % del ciclo de diseño. Las plataformas hardware reconfigurables, como son las FPGAs, también se benefician de las ventajas que ofrece la aritmética de coma fija, ya que éstas compensan las frecuencias de reloj más bajas y el uso más ineficiente del hardware que hacen estas plataformas respecto a los ASICs. A medida que las FPGAs se popularizan para su uso en computación científica los diseños aumentan de tamaño y complejidad hasta llegar al punto en que no pueden ser manejados eficientemente por las técnicas actuales de modelado de señal y ruido de cuantificación y de optimización de anchura de palabra. En esta Tesis Doctoral exploramos distintos aspectos del problema de la cuantificación y presentamos nuevas metodologías para cada uno de ellos: Las técnicas basadas en extensiones de intervalos han permitido obtener modelos de propagación de señal y ruido de cuantificación muy precisos en sistemas con operaciones no lineales. Nosotros llevamos esta aproximación un paso más allá introduciendo elementos de Multi-Element Generalized Polynomial Chaos (ME-gPC) y combinándolos con una técnica moderna basada en Modified Affine Arithmetic (MAA) estadístico para así modelar sistemas que contienen estructuras de control de flujo. Nuestra metodología genera los distintos caminos de ejecución automáticamente, determina las regiones del dominio de entrada que ejercitarán cada uno de ellos y extrae los momentos estadísticos del sistema a partir de dichas soluciones parciales. Utilizamos esta técnica para estimar tanto el rango dinámico como el ruido de redondeo en sistemas con las ya mencionadas estructuras de control de flujo y mostramos la precisión de nuestra aproximación, que en determinados casos de uso con operadores no lineales llega a tener tan solo una desviación del 0.04% con respecto a los valores de referencia obtenidos mediante simulación. Un inconveniente conocido de las técnicas basadas en extensiones de intervalos es la explosión combinacional de términos a medida que el tamaño de los sistemas a estudiar crece, lo cual conlleva problemas de escalabilidad. Para afrontar este problema presen tamos una técnica de inyección de ruidos agrupados que hace grupos con las señales del sistema, introduce las fuentes de ruido para cada uno de los grupos por separado y finalmente combina los resultados de cada uno de ellos. De esta forma, el número de fuentes de ruido queda controlado en cada momento y, debido a ello, la explosión combinatoria se minimiza. También presentamos un algoritmo de particionado multi-vía destinado a minimizar la desviación de los resultados a causa de la pérdida de correlación entre términos de ruido con el objetivo de mantener los resultados tan precisos como sea posible. La presente Tesis Doctoral también aborda el desarrollo de metodologías de optimización de anchura de palabra basadas en simulaciones de Monte-Cario que se ejecuten en tiempos razonables. Para ello presentamos dos nuevas técnicas que exploran la reducción del tiempo de ejecución desde distintos ángulos: En primer lugar, el método interpolativo aplica un interpolador sencillo pero preciso para estimar la sensibilidad de cada señal, y que es usado después durante la etapa de optimización. En segundo lugar, el método incremental gira en torno al hecho de que, aunque es estrictamente necesario mantener un intervalo de confianza dado para los resultados finales de nuestra búsqueda, podemos emplear niveles de confianza más relajados, lo cual deriva en un menor número de pruebas por simulación, en las etapas iniciales de la búsqueda, cuando todavía estamos lejos de las soluciones optimizadas. Mediante estas dos aproximaciones demostramos que podemos acelerar el tiempo de ejecución de los algoritmos clásicos de búsqueda voraz en factores de hasta x240 para problemas de tamaño pequeño/mediano. Finalmente, este libro presenta HOPLITE, una infraestructura de cuantificación automatizada, flexible y modular que incluye la implementación de las técnicas anteriores y se proporciona de forma pública. Su objetivo es ofrecer a desabolladores e investigadores un entorno común para prototipar y verificar nuevas metodologías de cuantificación de forma sencilla. Describimos el flujo de trabajo, justificamos las decisiones de diseño tomadas, explicamos su API pública y hacemos una demostración paso a paso de su funcionamiento. Además mostramos, a través de un ejemplo sencillo, la forma en que conectar nuevas extensiones a la herramienta con las interfaces ya existentes para poder así expandir y mejorar las capacidades de HOPLITE. ABSTRACT Using fixed-point arithmetic is one of the most common design choices for systems where area, power or throughput are heavily constrained. In order to produce implementations where the cost is minimized without negatively impacting the accuracy of the results, a careful assignment of word-lengths is required. The problem of finding the optimal combination of fixed-point word-lengths for a given system is a combinatorial NP-hard problem to which developers devote between 25 and 50% of the design-cycle time. Reconfigurable hardware platforms such as FPGAs also benefit of the advantages of fixed-point arithmetic, as it compensates for the slower clock frequencies and less efficient area utilization of the hardware platform with respect to ASICs. As FPGAs become commonly used for scientific computation, designs constantly grow larger and more complex, up to the point where they cannot be handled efficiently by current signal and quantization noise modelling and word-length optimization methodologies. In this Ph.D. Thesis we explore different aspects of the quantization problem and we present new methodologies for each of them: The techniques based on extensions of intervals have allowed to obtain accurate models of the signal and quantization noise propagation in systems with non-linear operations. We take this approach a step further by introducing elements of MultiElement Generalized Polynomial Chaos (ME-gPC) and combining them with an stateof- the-art Statistical Modified Affine Arithmetic (MAA) based methodology in order to model systems that contain control-flow structures. Our methodology produces the different execution paths automatically, determines the regions of the input domain that will exercise them, and extracts the system statistical moments from the partial results. We use this technique to estimate both the dynamic range and the round-off noise in systems with the aforementioned control-flow structures. We show the good accuracy of our approach, which in some case studies with non-linear operators shows a 0.04 % deviation respect to the simulation-based reference values. A known drawback of the techniques based on extensions of intervals is the combinatorial explosion of terms as the size of the targeted systems grows, which leads to scalability problems. To address this issue we present a clustered noise injection technique that groups the signals in the system, introduces the noise terms in each group independently and then combines the results at the end. In this way, the number of noise sources in the system at a given time is controlled and, because of this, the combinato rial explosion is minimized. We also present a multi-way partitioning algorithm aimed at minimizing the deviation of the results due to the loss of correlation between noise terms, in order to keep the results as accurate as possible. This Ph.D. Thesis also covers the development of methodologies for word-length optimization based on Monte-Carlo simulations in reasonable times. We do so by presenting two novel techniques that explore the reduction of the execution times approaching the problem in two different ways: First, the interpolative method applies a simple but precise interpolator to estimate the sensitivity of each signal, which is later used to guide the optimization effort. Second, the incremental method revolves on the fact that, although we strictly need to guarantee a certain confidence level in the simulations for the final results of the optimization process, we can do it with more relaxed levels, which in turn implies using a considerably smaller amount of samples, in the initial stages of the process, when we are still far from the optimized solution. Through these two approaches we demonstrate that the execution time of classical greedy techniques can be accelerated by factors of up to ×240 for small/medium sized problems. Finally, this book introduces HOPLITE, an automated, flexible and modular framework for quantization that includes the implementation of the previous techniques and is provided for public access. The aim is to offer a common ground for developers and researches for prototyping and verifying new techniques for system modelling and word-length optimization easily. We describe its work flow, justifying the taken design decisions, explain its public API and we do a step-by-step demonstration of its execution. We also show, through an example, the way new extensions to the flow should be connected to the existing interfaces in order to expand and improve the capabilities of HOPLITE.

A negative regulator mediates quorum-sensing control of exopolysaccharide production in Pantoea stewartii subsp. stewartii

Classical quorum-sensing (autoinduction) regulation, as exemplified by the lux system of Vibrio fischeri, requires N-acyl homoserine lactone (AHL) signals to stimulate cognate transcriptional activators for the cell density-dependent expression of specific target gene systems. For Pantoea stewartii subsp. stewartii, a bacterial pathogen of sweet corn and maize, the extracellular polysaccharide (EPS) stewartan is a major virulence factor, and its production is controlled by quorum sensing in a population density-dependent manner. Two genes, esaI and esaR, encode essential regulatory proteins for quorum sensing. EsaI is the AHL signal synthase, and EsaR is the cognate gene regulator. esaI, ΔesaR, and ΔesaI-esaR mutations were constructed to establish the regulatory role of EsaR. We report here that strains containing an esaR mutation produce high levels of EPS independently of cell density and in the absence of the AHL signal. Our data indicate that quorum-sensing regulation in P. s. subsp. stewartii, in contrast to most other described systems, uses EsaR to repress EPS synthesis at low cell density, and that derepression requires micromolar amounts of AHL. In addition, derepressed esaR strains, which synthesize EPS constitutively at low cell densities, were significantly less virulent than the wild-type parent. This finding suggests that quorum sensing in P. s. subsp. stewartii may be a mechanism to delay the expression of EPS during the early stages of infection so that it does not interfere with other mechanisms of pathogenesis.

A peptide export–import control circuit modulating bacterial development regulates protein phosphatases of the phosphorelay

The phosphorelay signal transduction system activates developmental transcription in sporulation of Bacillus subtilis by phosphorylation of aspartyl residues of the Spo0F and Spo0A response regulators. The phosphorylation level of these response regulators is determined by the opposing activities of protein kinases and protein aspartate phosphatases that interpret positive and negative signals for development in a signal integration circuit. The RapA protein aspartate phosphatase of the phosphorelay is regulated by a peptide that directly inhibits its activity. This peptide is proteolytically processed from an inactive pre-inhibitor protein encoded in the phrA gene. The pre-inhibitor is cleaved by the protein export apparatus to a putative pro-inhibitor that is further processed to the active inhibitor peptide and internalized by the oligopeptide permease. This export–import circuit is postulated to be a mechanism for timing phosphatase activity where the processing enzymes regulate the rate of formation of the active inhibitor. The processing events may, in turn, be controlled by a regulatory hierarchy. Chromosome sequencing has revealed several other phosphatase–prepeptide gene pairs in B. subtilis, suggesting that the use of this mechanism may be widespread in signal transduction.

Recognition of Yeast mRNAs as “Nonsense Containing” Leads to Both Inhibition of mRNA Translation and mRNA Degradation: Implications for the Control of mRNA Decapping

A critical step in the degradation of many eukaryotic mRNAs is a decapping reaction that exposes the transcript to 5′ to 3′ exonucleolytic degradation. The dual role of the cap structure as a target of mRNA degradation and as the site of assembly of translation initiation factors has led to the hypothesis that the rate of decapping would be specified by the status of the cap binding complex. This model makes the prediction that signals that promote mRNA decapping should also alter translation. To test this hypothesis, we examined the decapping triggered by premature termination codons to determine whether there is a down-regulation of translation when mRNAs were recognized as “nonsense containing.” We constructed an mRNA containing a premature stop codon in which we could measure the levels of both the mRNA and the polypeptide encoded upstream of the premature stop codon. Using this system, we analyzed the effects of premature stop codons on the levels of protein being produced per mRNA. In addition, by using alterations either in cis or in trans that inactivate different steps in the recognition and degradation of nonsense-containing mRNAs, we demonstrated that the recognition of a nonsense codon led to a decrease in the translational efficiency of the mRNA. These observations argue that the signal from a premature termination codon impinges on the translation machinery and suggest that decapping is a consequence of the change in translational status of the mRNA.

Mitochondrial control of calcium-channel gating: A mechanism for sustained signaling and transcriptional activation in T lymphocytes

In addition to their well-known functions in cellular energy transduction, mitochondria play an important role in modulating the amplitude and time course of intracellular Ca2+ signals. In many cells, mitochondria act as Ca2+ buffers by taking up and releasing Ca2+, but this simple buffering action by itself often cannot explain the organelle's effects on Ca2+ signaling dynamics. Here we describe the functional interaction of mitochondria with store-operated Ca2+ channels in T lymphocytes as a mechanism of mitochondrial Ca2+ signaling. In Jurkat T cells with functional mitochondria, prolonged depletion of Ca2+ stores causes sustained activation of the store-operated Ca2+ current, ICRAC (CRAC, Ca2+ release-activated Ca2+). Inhibition of mitochondrial Ca2+ uptake by compounds that dissipate the intramitochondrial potential unmasks Ca2+-dependent inactivation of ICRAC. Thus, functional mitochondria are required to maintain CRAC-channel activity, most likely by preventing local Ca2+ accumulation near sites that govern channel inactivation. In cells stimulated through the T-cell antigen receptor, acute blockade of mitochondrial Ca2+ uptake inhibits the nuclear translocation of the transcription factor NFAT in parallel with CRAC channel activity and [Ca2+]i elevation, indicating a functional link between mitochondrial regulation of ICRAC and T-cell activation. These results demonstrate a role for mitochondria in controlling Ca2+ channel activity and signal transmission from the plasma membrane to the nucleus.

Mechanism in the Sequential Control of Cell Morphology and S Phase Entry by Epidermal Growth Factor Involves Distinct MEK/ERK Activations

Cell shape plays a role in cell growth, differentiation, and death. Herein, we used the hepatocyte, a normal, highly differentiated cell characterized by a long G1 phase, to understand the mechanisms that link cell shape to growth. First, evidence was provided that the mitogen-activated protein kinase kinase (MEK)/extracellular signal-regulated kinase (ERK) cascade is a key transduction pathway controlling the hepatocyte morphology. MEK2/ERK2 activation in early G1 phase did not lead to cell proliferation but induced cell shape spreading and demonstration was provided that this MAPK-dependent spreading was required for reaching G1/S transition and DNA replication. Moreover, epidermal growth factor (EGF) was found to control this morphogenic signal in addition to its mitogenic effect. Thus, blockade of cell spreading by cytochalasin D or PD98059 treatment resulted in inhibition of EGF-dependent DNA replication. Our data led us to assess the first third of G1, is exclusively devoted to the growth factor-dependent morphogenic events, whereas the mitogenic signal occured at only approximately mid-G1 phase. Moreover, these two growth factor-related sequential signaling events involved successively activation of MEK2-ERK2 and then MEK1/2-ERK1/2 isoforms. In addition, we demonstrated that inhibition of extracellular matrix receptor, such as integrin β1 subunit, leads to cell arrest in G1, whereas EGF was found to up-regulated integrin β1 and fibronectin in a MEK-ERK–dependent manner. This process in relation to cytoskeletal reorganization could induce hepatocyte spreading, making them permissive for DNA replication. Our results provide new insight into the mechanisms by which a growth factor can temporally control dual morphogenic and mitogenic signals during the G1 phase.

Rac and Cdc42 GTPases control hematopoietic stem cell shape, adhesion, migration, and mobilization

Critical to homeostasis of blood cell production by hematopoietic stem/progenitor (HSC/P) cells is the regulation of HSC/P retention within the bone marrow microenvironment and migration between the bone marrow and the blood. Key extracellular regulatory elements for this process have been defined (cell–cell adhesion, growth factors, chemokines), but the mechanism by which HSC/P cells reconcile multiple external signals has not been elucidated. Rac and related small GTPases are candidates for this role and were studied in HSC/P deficient in Rac2, a hematopoietic cell-specific family member. Rac2 appears to be critical for HSC/P adhesion both in vitro and in vivo, whereas a compensatory increase in Cdc42 activation regulates HSC/P migration. This genetic analysis provides physiological evidence of cross-talk between GTPase proteins and suggests that a balance of these two GTPases controls HSC/P adhesion and mobilization in vivo.