Service needs forecasting: an approach for the automotive industry using analogies with medical ER management models.

During the past years, the industry has shifted position and moved towards “the luxury universe” whose customers are demanding, treating individuals as unique and valued customer for the business, offering vehicles produced with the state of the art technologies and implementing the highest finishing standards. Due to the competitive level in the market, car makers enable processes which equalizes customer services to E.R. management, being dealt with the maximum urgency that allows the comparison between both, car workshops and emergency rooms, where workshop bays or ramps will be equal to emergency boxes and skilled technicians are equivalent to the health care specialist, who will carry out tests and checks prior to afford any final operation, keeping the “patient” under control before it is back to normal utilization. This paper establishes a valid model for the automotive industry to estimate customer service demand forecasting under variable demand conditions using analogies with patient demand models used for the medical ER.

SERVICE NEEDS FORECASTING: An Approach for the Motor Industry Using Analogies with Medical ER Management Models.

During the past years, the industry has shifted position and moved towards “the luxury universe” whose customers are demanding, treating individuals as unique and valued customer for the business, offering vehicles produced with the state of the art technologies and implementing the highest finishing standards. Due to the competitive level in the market, motor makers enable processes which equalizes customer services to E.R. management, being dealt with the maximum urgency that allows the comparison between both, car workshops and emergency rooms, where workshop bays or ramps will be equal to emergency boxes and skilled technicians are equivalent to the health care specialist, who will carry out tests and checks prior to afford any final operation, keeping the “patient” under control before it is back to normal utilization. This paper ratify a valid model for the automotive industry to estimate customer service demand forecasting under variable demand conditions using analogies with patient demand models used for the medical ER

Driving assistance system applied in curves for real vehicles | Sistema de ayuda a la conducción en curvas para vehículos reales

Decreasing the accidents on highway and urban environments is the main motivation for the research and developing of driving assistance systems, also called ADAS (Advanced Driver Assistance Systems). In recent years, there are many applications of these systems in commercial vehicles: ABS systems, Cruise Control (CC), parking assistance and warning systems (including GPS), among others. However, the implementation of driving assistance systems on the steering wheel is more limited, because of their complexity and sensitivity. This paper is focused in the development, test and implementation of a driver assistance system for controlling the steering wheel in curve zones. This system is divided in two levels: an inner control loop which permits to execute the position and speed target, softening the action over the steering wheel, and a second control outer loop (controlling for fuzzy logic) that sends the reference to the inner loop according the environment and vehicle conditions. The tests have been done in different curves and speeds. The system has been proved in a commercial vehicle with satisfactory results.

An abstract machine based execution model for computer architecture design and efficient implementation of logic programs in parallel.

The term "Logic Programming" refers to a variety of computer languages and execution models which are based on the traditional concept of Symbolic Logic. The expressive power of these languages offers promise to be of great assistance in facing the programming challenges of present and future symbolic processing applications in Artificial Intelligence, Knowledge-based systems, and many other areas of computing. The sequential execution speed of logic programs has been greatly improved since the advent of the first interpreters. However, higher inference speeds are still required in order to meet the demands of applications such as those contemplated for next generation computer systems. The execution of logic programs in parallel is currently considered a promising strategy for attaining such inference speeds. Logic Programming in turn appears as a suitable programming paradigm for parallel architectures because of the many opportunities for parallel execution present in the implementation of logic programs. This dissertation presents an efficient parallel execution model for logic programs. The model is described from the source language level down to an "Abstract Machine" level suitable for direct implementation on existing parallel systems or for the design of special purpose parallel architectures. Few assumptions are made at the source language level and therefore the techniques developed and the general Abstract Machine design are applicable to a variety of logic (and also functional) languages. These techniques offer efficient solutions to several areas of parallel Logic Programming implementation previously considered problematic or a source of considerable overhead, such as the detection and handling of variable binding conflicts in AND-Parallelism, the specification of control and management of the execution tree, the treatment of distributed backtracking, and goal scheduling and memory management issues, etc. A parallel Abstract Machine design is offered, specifying data areas, operation, and a suitable instruction set. This design is based on extending to a parallel environment the techniques introduced by the Warren Abstract Machine, which have already made very fast and space efficient sequential systems a reality. Therefore, the model herein presented is capable of retaining sequential execution speed similar to that of high performance sequential systems, while extracting additional gains in speed by efficiently implementing parallel execution. These claims are supported by simulations of the Abstract Machine on sample programs.

Caracterización de un robot para aplicaciones de mecanizado con requerimientos de tolerancias

El presente Trabajo fin Fin de Máster, versa sobre una caracterización preliminar del comportamiento de un robot de tipo industrial, configurado por 4 eslabones y 4 grados de libertad, y sometido a fuerzas de mecanizado en su extremo. El entorno de trabajo planteado es el de plantas de fabricación de piezas de aleaciones de aluminio para automoción. Este tipo de componentes parte de un primer proceso de fundición que saca la pieza en bruto. Para series medias y altas, en función de las propiedades mecánicas y plásticas requeridas y los costes de producción, la inyección a alta presión (HPDC) y la fundición a baja presión (LPC) son las dos tecnologías más usadas en esta primera fase. Para inyección a alta presión, las aleaciones de aluminio más empleadas son, en designación simbólica según norma EN 1706 (entre paréntesis su designación numérica); EN AC AlSi9Cu3(Fe) (EN AC 46000) , EN AC AlSi9Cu3(Fe)(Zn) (EN AC 46500), y EN AC AlSi12Cu1(Fe) (EN AC 47100). Para baja presión, EN AC AlSi7Mg0,3 (EN AC 42100). En los 3 primeros casos, los límites de Silicio permitidos pueden superan el 10%. En el cuarto caso, es inferior al 10% por lo que, a los efectos de ser sometidas a mecanizados, las piezas fabricadas en aleaciones con Si superior al 10%, se puede considerar que son equivalentes, diferenciándolas de la cuarta. Las tolerancias geométricas y dimensionales conseguibles directamente de fundición, recogidas en normas como ISO 8062 o DIN 1688-1, establecen límites para este proceso. Fuera de esos límites, las garantías en conseguir producciones con los objetivos de ppms aceptados en la actualidad por el mercado, obligan a ir a fases posteriores de mecanizado. Aquellas geometrías que, funcionalmente, necesitan disponer de unas tolerancias geométricas y/o dimensionales definidas acorde a ISO 1101, y no capaces por este proceso inicial de moldeado a presión, deben ser procesadas en una fase posterior en células de mecanizado. En este caso, las tolerancias alcanzables para procesos de arranque de viruta se recogen en normas como ISO 2768. Las células de mecanizado se componen, por lo general, de varios centros de control numérico interrelacionados y comunicados entre sí por robots que manipulan las piezas en proceso de uno a otro. Dichos robots, disponen en su extremo de una pinza utillada para poder coger y soltar las piezas en los útiles de mecanizado, las mesas de intercambio para cambiar la pieza de posición o en utillajes de equipos de medición y prueba, o en cintas de entrada o salida. La repetibilidad es alta, de centésimas incluso, definida según norma ISO 9283. El problema es que, estos rangos de repetibilidad sólo se garantizan si no se hacen esfuerzos o éstos son despreciables (caso de mover piezas). Aunque las inercias de mover piezas a altas velocidades hacen que la trayectoria intermedia tenga poca precisión, al inicio y al final (al coger y dejar pieza, p.e.) se hacen a velocidades relativamente bajas que hacen que el efecto de las fuerzas de inercia sean menores y que permiten garantizar la repetibilidad anteriormente indicada. No ocurre así si se quitara la garra y se intercambia con un cabezal motorizado con una herramienta como broca, mandrino, plato de cuchillas, fresas frontales o tangenciales… Las fuerzas ejercidas de mecanizado generarían unos pares en las uniones tan grandes y tan variables que el control del robot no sería capaz de responder (o no está preparado, en un principio) y generaría una desviación en la trayectoria, realizada a baja velocidad, que desencadenaría en un error de posición (ver norma ISO 5458) no asumible para la funcionalidad deseada. Se podría llegar al caso de que la tolerancia alcanzada por un pretendido proceso más exacto diera una dimensión peor que la que daría el proceso de fundición, en principio con mayor variabilidad dimensional en proceso (y por ende con mayor intervalo de tolerancia garantizable). De hecho, en los CNCs, la precisión es muy elevada, (pudiéndose despreciar en la mayoría de los casos) y no es la responsable de, por ejemplo la tolerancia de posición al taladrar un agujero. Factores como, temperatura de la sala y de la pieza, calidad constructiva de los utillajes y rigidez en el amarre, error en el giro de mesas y de colocación de pieza, si lleva agujeros previos o no, si la herramienta está bien equilibrada y el cono es el adecuado para el tipo de mecanizado… influyen más. Es interesante que, un elemento no específico tan común en una planta industrial, en el entorno anteriormente descrito, como es un robot, el cual no sería necesario añadir por disponer de él ya (y por lo tanto la inversión sería muy pequeña), puede mejorar la cadena de valor disminuyendo el costo de fabricación. Y si se pudiera conjugar que ese robot destinado a tareas de manipulación, en los muchos tiempos de espera que va a disfrutar mientras el CNC arranca viruta, pudiese coger un cabezal y apoyar ese mecanizado; sería doblemente interesante. Por lo tanto, se antoja sugestivo poder conocer su comportamiento e intentar explicar qué sería necesario para llevar esto a cabo, motivo de este trabajo. La arquitectura de robot seleccionada es de tipo SCARA. La búsqueda de un robot cómodo de modelar y de analizar cinemática y dinámicamente, sin limitaciones relevantes en la multifuncionalidad de trabajos solicitados, ha llevado a esta elección, frente a otras arquitecturas como por ejemplo los robots antropomórficos de 6 grados de libertad, muy populares a nivel industrial. Este robot dispone de 3 uniones, de las cuales 2 son de tipo par de revolución (1 grado de libertad cada una) y la tercera es de tipo corredera o par cilíndrico (2 grados de libertad). La primera unión, de tipo par de revolución, sirve para unir el suelo (considerado como eslabón número 1) con el eslabón número 2. La segunda unión, también de ese tipo, une el eslabón número 2 con el eslabón número 3. Estos 2 brazos, pueden describir un movimiento horizontal, en el plano X-Y. El tercer eslabón, está unido al eslabón número 4 por la unión de tipo corredera. El movimiento que puede describir es paralelo al eje Z. El robot es de 4 grados de libertad (4 motores). En relación a los posibles trabajos que puede realizar este tipo de robot, su versatilidad abarca tanto operaciones típicas de manipulación como operaciones de arranque de viruta. Uno de los mecanizados más usuales es el taladrado, por lo cual se elige éste para su modelización y análisis. Dentro del taladrado se elegirá para acotar las fuerzas, taladrado en macizo con broca de diámetro 9 mm. El robot se ha considerado por el momento que tenga comportamiento de sólido rígido, por ser el mayor efecto esperado el de los pares en las uniones. Para modelar el robot se utiliza el método de los sistemas multicuerpos. Dentro de este método existen diversos tipos de formulaciones (p.e. Denavit-Hartenberg). D-H genera una cantidad muy grande de ecuaciones e incógnitas. Esas incógnitas son de difícil comprensión y, para cada posición, hay que detenerse a pensar qué significado tienen. Se ha optado por la formulación de coordenadas naturales. Este sistema utiliza puntos y vectores unitarios para definir la posición de los distintos cuerpos, y permite compartir, cuando es posible y se quiere, para definir los pares cinemáticos y reducir al mismo tiempo el número de variables. Las incógnitas son intuitivas, las ecuaciones de restricción muy sencillas y se reduce considerablemente el número de ecuaciones e incógnitas. Sin embargo, las coordenadas naturales “puras” tienen 2 problemas. El primero, que 2 elementos con un ángulo de 0 o 180 grados, dan lugar a puntos singulares que pueden crear problemas en las ecuaciones de restricción y por lo tanto han de evitarse. El segundo, que tampoco inciden directamente sobre la definición o el origen de los movimientos. Por lo tanto, es muy conveniente complementar esta formulación con ángulos y distancias (coordenadas relativas). Esto da lugar a las coordenadas naturales mixtas, que es la formulación final elegida para este TFM. Las coordenadas naturales mixtas no tienen el problema de los puntos singulares. Y la ventaja más importante reside en su utilidad a la hora de aplicar fuerzas motrices, momentos o evaluar errores. Al incidir sobre la incógnita origen (ángulos o distancias) controla los motores de manera directa. El algoritmo, la simulación y la obtención de resultados se ha programado mediante Matlab. Para realizar el modelo en coordenadas naturales mixtas, es preciso modelar en 2 pasos el robot a estudio. El primer modelo se basa en coordenadas naturales. Para su validación, se plantea una trayectoria definida y se analiza cinemáticamente si el robot satisface el movimiento solicitado, manteniendo su integridad como sistema multicuerpo. Se cuantifican los puntos (en este caso inicial y final) que configuran el robot. Al tratarse de sólidos rígidos, cada eslabón queda definido por sus respectivos puntos inicial y final (que son los más interesantes para la cinemática y la dinámica) y por un vector unitario no colineal a esos 2 puntos. Los vectores unitarios se colocan en los lugares en los que se tenga un eje de rotación o cuando se desee obtener información de un ángulo. No son necesarios vectores unitarios para medir distancias. Tampoco tienen por qué coincidir los grados de libertad con el número de vectores unitarios. Las longitudes de cada eslabón quedan definidas como constantes geométricas. Se establecen las restricciones que definen la naturaleza del robot y las relaciones entre los diferentes elementos y su entorno. La trayectoria se genera por una nube de puntos continua, definidos en coordenadas independientes. Cada conjunto de coordenadas independientes define, en un instante concreto, una posición y postura de robot determinada. Para conocerla, es necesario saber qué coordenadas dependientes hay en ese instante, y se obtienen resolviendo por el método de Newton-Rhapson las ecuaciones de restricción en función de las coordenadas independientes. El motivo de hacerlo así es porque las coordenadas dependientes deben satisfacer las restricciones, cosa que no ocurre con las coordenadas independientes. Cuando la validez del modelo se ha probado (primera validación), se pasa al modelo 2. El modelo número 2, incorpora a las coordenadas naturales del modelo número 1, las coordenadas relativas en forma de ángulos en los pares de revolución (3 ángulos; ϕ1, ϕ 2 y ϕ3) y distancias en los pares prismáticos (1 distancia; s). Estas coordenadas relativas pasan a ser las nuevas coordenadas independientes (sustituyendo a las coordenadas independientes cartesianas del modelo primero, que eran coordenadas naturales). Es necesario revisar si el sistema de vectores unitarios del modelo 1 es suficiente o no. Para este caso concreto, se han necesitado añadir 1 vector unitario adicional con objeto de que los ángulos queden perfectamente determinados con las correspondientes ecuaciones de producto escalar y/o vectorial. Las restricciones habrán de ser incrementadas en, al menos, 4 ecuaciones; una por cada nueva incógnita. La validación del modelo número 2, tiene 2 fases. La primera, al igual que se hizo en el modelo número 1, a través del análisis cinemático del comportamiento con una trayectoria definida. Podrían obtenerse del modelo 2 en este análisis, velocidades y aceleraciones, pero no son necesarios. Tan sólo interesan los movimientos o desplazamientos finitos. Comprobada la coherencia de movimientos (segunda validación), se pasa a analizar cinemáticamente el comportamiento con trayectorias interpoladas. El análisis cinemático con trayectorias interpoladas, trabaja con un número mínimo de 3 puntos máster. En este caso se han elegido 3; punto inicial, punto intermedio y punto final. El número de interpolaciones con el que se actúa es de 50 interpolaciones en cada tramo (cada 2 puntos máster hay un tramo), resultando un total de 100 interpolaciones. El método de interpolación utilizado es el de splines cúbicas con condición de aceleración inicial y final constantes, que genera las coordenadas independientes de los puntos interpolados de cada tramo. Las coordenadas dependientes se obtienen resolviendo las ecuaciones de restricción no lineales con el método de Newton-Rhapson. El método de las splines cúbicas es muy continuo, por lo que si se desea modelar una trayectoria en el que haya al menos 2 movimientos claramente diferenciados, es preciso hacerlo en 2 tramos y unirlos posteriormente. Sería el caso en el que alguno de los motores se desee expresamente que esté parado durante el primer movimiento y otro distinto lo esté durante el segundo movimiento (y así sucesivamente). Obtenido el movimiento, se calculan, también mediante fórmulas de diferenciación numérica, las velocidades y aceleraciones independientes. El proceso es análogo al anteriormente explicado, recordando la condición impuesta de que la aceleración en el instante t= 0 y en instante t= final, se ha tomado como 0. Las velocidades y aceleraciones dependientes se calculan resolviendo las correspondientes derivadas de las ecuaciones de restricción. Se comprueba, de nuevo, en una tercera validación del modelo, la coherencia del movimiento interpolado. La dinámica inversa calcula, para un movimiento definido -conocidas la posición, velocidad y la aceleración en cada instante de tiempo-, y conocidas las fuerzas externas que actúan (por ejemplo el peso); qué fuerzas hay que aplicar en los motores (donde hay control) para que se obtenga el citado movimiento. En la dinámica inversa, cada instante del tiempo es independiente de los demás y tiene una posición, una velocidad y una aceleración y unas fuerzas conocidas. En este caso concreto, se desean aplicar, de momento, sólo las fuerzas debidas al peso, aunque se podrían haber incorporado fuerzas de otra naturaleza si se hubiese deseado. Las posiciones, velocidades y aceleraciones, proceden del cálculo cinemático. El efecto inercial de las fuerzas tenidas en cuenta (el peso) es calculado. Como resultado final del análisis dinámico inverso, se obtienen los pares que han de ejercer los cuatro motores para replicar el movimiento prescrito con las fuerzas que estaban actuando. La cuarta validación del modelo consiste en confirmar que el movimiento obtenido por aplicar los pares obtenidos en la dinámica inversa, coinciden con el obtenido en el análisis cinemático (movimiento teórico). Para ello, es necesario acudir a la dinámica directa. La dinámica directa se encarga de calcular el movimiento del robot, resultante de aplicar unos pares en motores y unas fuerzas en el robot. Por lo tanto, el movimiento real resultante, al no haber cambiado ninguna condición de las obtenidas en la dinámica inversa (pares de motor y fuerzas inerciales debidas al peso de los eslabones) ha de ser el mismo al movimiento teórico. Siendo así, se considera que el robot está listo para trabajar. Si se introduce una fuerza exterior de mecanizado no contemplada en la dinámica inversa y se asigna en los motores los mismos pares resultantes de la resolución del problema dinámico inverso, el movimiento real obtenido no es igual al movimiento teórico. El control de lazo cerrado se basa en ir comparando el movimiento real con el deseado e introducir las correcciones necesarias para minimizar o anular las diferencias. Se aplican ganancias en forma de correcciones en posición y/o velocidad para eliminar esas diferencias. Se evalúa el error de posición como la diferencia, en cada punto, entre el movimiento teórico deseado en el análisis cinemático y el movimiento real obtenido para cada fuerza de mecanizado y una ganancia concreta. Finalmente, se mapea el error de posición obtenido para cada fuerza de mecanizado y las diferentes ganancias previstas, graficando la mejor precisión que puede dar el robot para cada operación que se le requiere, y en qué condiciones. -------------- This Master´s Thesis deals with a preliminary characterization of the behaviour for an industrial robot, configured with 4 elements and 4 degrees of freedoms, and subjected to machining forces at its end. Proposed working conditions are those typical from manufacturing plants with aluminium alloys for automotive industry. This type of components comes from a first casting process that produces rough parts. For medium and high volumes, high pressure die casting (HPDC) and low pressure die casting (LPC) are the most used technologies in this first phase. For high pressure die casting processes, most used aluminium alloys are, in simbolic designation according EN 1706 standard (between brackets, its numerical designation); EN AC AlSi9Cu3(Fe) (EN AC 46000) , EN AC AlSi9Cu3(Fe)(Zn) (EN AC 46500), y EN AC AlSi12Cu1(Fe) (EN AC 47100). For low pressure, EN AC AlSi7Mg0,3 (EN AC 42100). For the 3 first alloys, Si allowed limits can exceed 10% content. Fourth alloy has admisible limits under 10% Si. That means, from the point of view of machining, that components made of alloys with Si content above 10% can be considered as equivalent, and the fourth one must be studied separately. Geometrical and dimensional tolerances directly achievables from casting, gathered in standards such as ISO 8062 or DIN 1688-1, establish a limit for this process. Out from those limits, guarantees to achieve batches with objetive ppms currently accepted by market, force to go to subsequent machining process. Those geometries that functionally require a geometrical and/or dimensional tolerance defined according ISO 1101, not capable with initial moulding process, must be obtained afterwards in a machining phase with machining cells. In this case, tolerances achievables with cutting processes are gathered in standards such as ISO 2768. In general terms, machining cells contain several CNCs that they are interrelated and connected by robots that handle parts in process among them. Those robots have at their end a gripper in order to take/remove parts in machining fixtures, in interchange tables to modify position of part, in measurement and control tooling devices, or in entrance/exit conveyors. Repeatibility for robot is tight, even few hundredths of mm, defined according ISO 9283. Problem is like this; those repeatibilty ranks are only guaranteed when there are no stresses or they are not significant (f.e. due to only movement of parts). Although inertias due to moving parts at a high speed make that intermediate paths have little accuracy, at the beginning and at the end of trajectories (f.e, when picking part or leaving it) movement is made with very slow speeds that make lower the effect of inertias forces and allow to achieve repeatibility before mentioned. It does not happens the same if gripper is removed and it is exchanged by an spindle with a machining tool such as a drilling tool, a pcd boring tool, a face or a tangential milling cutter… Forces due to machining would create such big and variable torques in joints that control from the robot would not be able to react (or it is not prepared in principle) and would produce a deviation in working trajectory, made at a low speed, that would trigger a position error (see ISO 5458 standard) not assumable for requested function. Then it could be possible that tolerance achieved by a more exact expected process would turn out into a worst dimension than the one that could be achieved with casting process, in principle with a larger dimensional variability in process (and hence with a larger tolerance range reachable). As a matter of fact, accuracy is very tight in CNC, (its influence can be ignored in most cases) and it is not the responsible of, for example position tolerance when drilling a hole. Factors as, room and part temperature, manufacturing quality of machining fixtures, stiffness at clamping system, rotating error in 4th axis and part positioning error, if there are previous holes, if machining tool is properly balanced, if shank is suitable for that machining type… have more influence. It is interesting to know that, a non specific element as common, at a manufacturing plant in the enviroment above described, as a robot (not needed to be added, therefore with an additional minimum investment), can improve value chain decreasing manufacturing costs. And when it would be possible to combine that the robot dedicated to handling works could support CNCs´ works in its many waiting time while CNCs cut, and could take an spindle and help to cut; it would be double interesting. So according to all this, it would be interesting to be able to know its behaviour and try to explain what would be necessary to make this possible, reason of this work. Selected robot architecture is SCARA type. The search for a robot easy to be modeled and kinematically and dinamically analyzed, without significant limits in the multifunctionality of requested operations, has lead to this choice. Due to that, other very popular architectures in the industry, f.e. 6 DOFs anthropomorphic robots, have been discarded. This robot has 3 joints, 2 of them are revolute joints (1 DOF each one) and the third one is a cylindrical joint (2 DOFs). The first joint, a revolute one, is used to join floor (body 1) with body 2. The second one, a revolute joint too, joins body 2 with body 3. These 2 bodies can move horizontally in X-Y plane. Body 3 is linked to body 4 with a cylindrical joint. Movement that can be made is paralell to Z axis. The robt has 4 degrees of freedom (4 motors). Regarding potential works that this type of robot can make, its versatility covers either typical handling operations or cutting operations. One of the most common machinings is to drill. That is the reason why it has been chosen for the model and analysis. Within drilling, in order to enclose spectrum force, a typical solid drilling with 9 mm diameter. The robot is considered, at the moment, to have a behaviour as rigid body, as biggest expected influence is the one due to torques at joints. In order to modelize robot, it is used multibodies system method. There are under this heading different sorts of formulations (f.e. Denavit-Hartenberg). D-H creates a great amount of equations and unknown quantities. Those unknown quatities are of a difficult understanding and, for each position, one must stop to think about which meaning they have. The choice made is therefore one of formulation in natural coordinates. This system uses points and unit vectors to define position of each different elements, and allow to share, when it is possible and wished, to define kinematic torques and reduce number of variables at the same time. Unknown quantities are intuitive, constrain equations are easy and number of equations and variables are strongly reduced. However, “pure” natural coordinates suffer 2 problems. The first one is that 2 elements with an angle of 0° or 180°, give rise to singular positions that can create problems in constrain equations and therefore they must be avoided. The second problem is that they do not work directly over the definition or the origin of movements. Given that, it is highly recommended to complement this formulation with angles and distances (relative coordinates). This leads to mixed natural coordinates, and they are the final formulation chosen for this MTh. Mixed natural coordinates have not the problem of singular positions. And the most important advantage lies in their usefulness when applying driving forces, torques or evaluating errors. As they influence directly over origin variable (angles or distances), they control motors directly. The algorithm, simulation and obtaining of results has been programmed with Matlab. To design the model in mixed natural coordinates, it is necessary to model the robot to be studied in 2 steps. The first model is based in natural coordinates. To validate it, it is raised a defined trajectory and it is kinematically analyzed if robot fulfils requested movement, keeping its integrity as multibody system. The points (in this case starting and ending points) that configure the robot are quantified. As the elements are considered as rigid bodies, each of them is defined by its respectively starting and ending point (those points are the most interesting ones from the point of view of kinematics and dynamics) and by a non-colinear unit vector to those points. Unit vectors are placed where there is a rotating axis or when it is needed information of an angle. Unit vectors are not needed to measure distances. Neither DOFs must coincide with the number of unit vectors. Lengths of each arm are defined as geometrical constants. The constrains that define the nature of the robot and relationships among different elements and its enviroment are set. Path is generated by a cloud of continuous points, defined in independent coordinates. Each group of independent coordinates define, in an specific instant, a defined position and posture for the robot. In order to know it, it is needed to know which dependent coordinates there are in that instant, and they are obtained solving the constraint equations with Newton-Rhapson method according to independent coordinates. The reason to make it like this is because dependent coordinates must meet constraints, and this is not the case with independent coordinates. When suitability of model is checked (first approval), it is given next step to model 2. Model 2 adds to natural coordinates from model 1, the relative coordinates in the shape of angles in revoluting torques (3 angles; ϕ1, ϕ 2 and ϕ3) and distances in prismatic torques (1 distance; s). These relative coordinates become the new independent coordinates (replacing to cartesian independent coordinates from model 1, that they were natural coordinates). It is needed to review if unit vector system from model 1 is enough or not . For this specific case, it was necessary to add 1 additional unit vector to define perfectly angles with their related equations of dot and/or cross product. Constrains must be increased in, at least, 4 equations; one per each new variable. The approval of model 2 has two phases. The first one, same as made with model 1, through kinematic analysis of behaviour with a defined path. During this analysis, it could be obtained from model 2, velocities and accelerations, but they are not needed. They are only interesting movements and finite displacements. Once that the consistence of movements has been checked (second approval), it comes when the behaviour with interpolated trajectories must be kinematically analyzed. Kinematic analysis with interpolated trajectories work with a minimum number of 3 master points. In this case, 3 points have been chosen; starting point, middle point and ending point. The number of interpolations has been of 50 ones in each strecht (each 2 master points there is an strecht), turning into a total of 100 interpolations. The interpolation method used is the cubic splines one with condition of constant acceleration both at the starting and at the ending point. This method creates the independent coordinates of interpolated points of each strecht. The dependent coordinates are achieved solving the non-linear constrain equations with Newton-Rhapson method. The method of cubic splines is very continuous, therefore when it is needed to design a trajectory in which there are at least 2 movements clearly differents, it is required to make it in 2 steps and join them later. That would be the case when any of the motors would keep stopped during the first movement, and another different motor would remain stopped during the second movement (and so on). Once that movement is obtained, they are calculated, also with numerical differenciation formulas, the independent velocities and accelerations. This process is analogous to the one before explained, reminding condition that acceleration when t=0 and t=end are 0. Dependent velocities and accelerations are calculated solving related derivatives of constrain equations. In a third approval of the model it is checked, again, consistence of interpolated movement. Inverse dynamics calculates, for a defined movement –knowing position, velocity and acceleration in each instant of time-, and knowing external forces that act (f.e. weights); which forces must be applied in motors (where there is control) in order to obtain requested movement. In inverse dynamics, each instant of time is independent of the others and it has a position, a velocity, an acceleration and known forces. In this specific case, it is intended to apply, at the moment, only forces due to the weight, though forces of another nature could have been added if it would have been preferred. The positions, velocities and accelerations, come from kinematic calculation. The inertial effect of forces taken into account (weight) is calculated. As final result of the inverse dynamic analysis, the are obtained torques that the 4 motors must apply to repeat requested movement with the forces that were acting. The fourth approval of the model consists on confirming that the achieved movement due to the use of the torques obtained in the inverse dynamics, are in accordance with movements from kinematic analysis (theoretical movement). For this, it is necessary to work with direct dynamics. Direct dynamic is in charge of calculating the movements of robot that results from applying torques at motors and forces at the robot. Therefore, the resultant real movement, as there was no change in any condition of the ones obtained at the inverse dynamics (motor torques and inertial forces due to weight of elements) must be the same than theoretical movement. When these results are achieved, it is considered that robot is ready to work. When a machining external force is introduced and it was not taken into account before during the inverse dynamics, and torques at motors considered are the ones of the inverse dynamics, the real movement obtained is not the same than the theoretical movement. Closed loop control is based on comparing real movement with expected movement and introducing required corrrections to minimize or cancel differences. They are applied gains in the way of corrections for position and/or tolerance to remove those differences. Position error is evaluated as the difference, in each point, between theoretical movemment (calculated in the kinematic analysis) and the real movement achieved for each machining force and for an specific gain. Finally, the position error obtained for each machining force and gains are mapped, giving a chart with the best accuracy that the robot can give for each operation that has been requested and which conditions must be provided.

Functional response of Ulmus minor Mill. to drought, flooding and dutch elm disease

Ulmus minor es una especie arbórea originaria de Europa cuyas poblaciones han sido diezmadas por el hongo patógeno causante de la enfermedad de la grafiosis. La conservación de los olmos exige plantearse su propagación a través de plantaciones y conocer mejor su ecología y biología. Ulmus minor es un árbol de ribera, pero frecuentemente se encuentra alejado del cauce de arroyos y ríos, donde la capa freática sufre fuertes oscilaciones. Por ello, nuestra hipótesis general es que esta especie es moderadamente resistente tanto a la inundación como a la sequía. El principal objetivo de esta tesis doctoral es entender desde un punto de vista funcional la respuesta de U. minor a la inundación, la sequía y la infección por O. novo-ulmi; los factores que posiblemente más influyen en la distribución actual de U. minor. Con este objetivo se persigue dar continuidad a los esfuerzos de conservación de esta especie que desde hace años se dedican en varios centros de investigación a nivel mundial, ya que, entender mejor los mecanismos que contribuyen a la resistencia de U. minor ante la inoculación con O. novo-ulmi y factores de estrés abiótico ayudará en la selección y propagación de genotipos resistentes a la grafiosis. Se han planteado tres experimentos en este sentido. Primero, se ha comparado la tolerancia de brinzales de U. minor y U. laevis – otro olmo ibérico – a una inmersión controlada con el fin de evaluar su tolerancia a la inundación y comprender los mecanismos de aclimatación. Segundo, se ha comparado la tolerancia de brinzales de U. minor y Quercus ilex – una especie típica de ambientes Mediterránea secos – a la falta de agua en el suelo con el fin de evaluar el grado de tolerancia y los mecanismos de aclimatación a la sequía. El hecho de comparar dos especies contrastadas responde al interés en entender mejor cuales son los procesos que conducen a la muerte de una planta en condiciones de sequía – asunto sobre el que hay una interesante discusión desde hace algunos años. En tercer lugar, con el fin de entender mejor la resistencia de algunos genotipos de U. minor a la grafiosis, se han estudiado las diferencias fisiológicas y químicas constitutivas e inducidas por O. novo-ulmi entre clones de U. minor seleccionados a priori por su variable grado de resistencia a esta enfermedad. En el primer experimento se observó que los brinzales de U. minor sobrevivieron 60 días inmersos en una piscina con agua no estancada hasta una altura de 2-3 cm por encima del cuello de la raíz. A los 60 días, los brinzales de U. laevis se sacaron de la piscina y, a lo largo de las siguientes semanas, fueron capaces de recuperar las funciones fisiológicas que habían sido alteradas anteriormente. La conductividad hidráulica de las raíces y la tasa de asimilación de CO2 neta disminuyeron en ambas especies. Por el contrario, la tasa de respiración de hojas, tallos y raíces aumentó en las primeras semanas de la inundación, posiblemente en relación al aumento de energía necesario para desarrollar mecanismos de aclimatación a la inundación, como la hipertrofia de las lenticelas que se observó en ambas especies. Por ello, el desequilibrio del balance de carbono de la planta podría ser un factor relevante en la mortalidad de las plantas ante inundaciones prolongadas. Las plantas de U. minor (cultivadas en envases de 16 litros a media sombra) sobrevivieron por un prolongado periodo de tiempo en verano sin riego; la mitad de las plantas murieron tras 90 días sin riego. El cierre de los estomas y la pérdida de hojas contribuyeron a ralentizar las pérdidas de agua y tolerar la sequía en U. minor. Las obvias diferencias en tolerancia a la sequía con respecto a Q. ilex se reflejaron en la distinta capacidad para ralentizar la aparición del estrés hídrico tras dejar de regar y para transportar agua en condiciones de elevada tensión en el xilema. Más relevante es que las plantas con evidentes síntomas de decaimiento previo a su muerte exhibieron pérdidas de conductividad hidráulica en las raíces del 80% en ambas especies, mientras que las reservas de carbohidratos apenas variaron y lo hicieron de forma desigual en ambas especies. Árboles de U. minor de 5 y 6 años de edad (plantados en eras con riego mantenido) exhibieron una respuesta a la inoculación con O. novo-ulmi consistente con ensayos previos de resistencia. La conductividad hidráulica del tallo, el potencial hídrico foliar y la tasa de asimilación de CO2 neta disminuyeron significativamente en relación a árboles inoculados con agua, pero solo en los clones susceptibles. Este hecho enlaza con el perfil químico “más defensivo” de los clones resistentes, es decir, con los mayores niveles de suberina, ácidos grasos y compuestos fenólicos en estos clones que en los susceptibles. Ello podría restringir la propagación del hongo en el árbol y preservar el comportamiento fisiológico de los clones resistentes al inocularlos con el patógeno. Los datos indican una respuesta fisiológica común de U. minor a la inundación, la sequía y la infección por O. novo-ulmi: pérdida de conductividad hidráulica, estrés hídrico y pérdida de ganancia neta de carbono. Pese a ello, U. minor desarrolla varios mecanismos que le confieren una capacidad moderada para vivir en suelos temporalmente anegados o secos. Por otro lado, el perfil químico es un factor relevante en la resistencia de ciertos genotipos a la grafiosis. Futuros estudios deberían examinar como este perfil químico y la resistencia a la grafiosis se ven alteradas por el estrés abiótico. ABSTRACT Ulmus minor is a native European elm species whose populations have been decimated by the Dutch elm disease (DED). An active conservation of this species requires large-scale plantations and a better understanding of its biology and ecology. U. minor generally grows close to water channels. However, of the Iberian riparian tree species, U. minor is the one that spread farther away from rivers and streams. For these reasons, we hypothesize that this species is moderately tolerant to both flooding and drought stresses. The main aim of the present PhD thesis is to better understand the functional response of U. minor to the abiotic stresses – flooding and drought – and the biotic stress – DED – that can be most influential on its distribution. The overarching goal is to aid in the conservation of this emblematic species through a better understanding of the mechanisms that contribute to resistance to abiotic and biotic stresses; an information that can help in the selection of resistant genotypes and their expansion in large-scale plantations. To this end, three experiments were set up. First, we compared the tolerance to experimental immersion between seedlings of U. minor and U. laevis – another European riparian elm species – in order to assess their degree of tolerance and understand the mechanisms of acclimation to this stress. Second, we investigated the tolerance to drought of U. minor seedlings in comparison with Quercus ilex (an oak species typical of dry Mediterranean habitats). Besides assessing and understanding U. minor tolerance to drought at the seedling stage, the aim was to shed light into the functional alterations that trigger drought-induced plant mortality – a matter of controversy in the last years. Third, we studied constitutive and induced physiological and biochemical differences among clones of variable DED resistance, before and following inoculation with Ophiostoma novo-ulmi. The goal is to shed light into the factors of DED resistance that is evident in some genotypes of U. minor, but not others. Potted seedlings of U. minor survived for 60 days immersed in a pool with running water to approximately 2-3 cm above the stem collar. By this time, U. minor seedlings died, whereas U. laevis seedlings moved out of the pool were able to recover most physiological functions that had been altered by flooding. For example, root hydraulic conductivity and leaf photosynthetic CO2 uptake decreased in both species; while respiration initially increased with flooding in leaves, stems and roots possibly to respond to energy demands associated to mechanisms of acclimation to soil oxygen deficiency; as example, a remarkable hypertrophy of lenticels was soon observed in flooded seedlings of both species. Therefore, the inability to maintain a positive carbon balance somehow compromises seedling survival under flooding, earlier in U. minor than U. laevis, partly explaining their differential habitats. Potted seedlings of U. minor survived for a remarkable long time without irrigation – half of plants dying only after 90 days of no irrigation in conditions of high vapour pressure deficit typical of summer. Some mechanisms that contributed to tolerate drought were leaf shedding and stomata closure, which reduced water loss and the risk of xylem cavitation. Obviously, U. minor was less tolerant to drought than Q. ilex, differences in drought tolerance resulting mostly from the distinct capacity to postpone water stress and conduct water under high xylem tension among species. More relevant was that plants of both species exhibited similar symptoms of root hydraulic failure (i.e. approximately 80% loss of hydraulic conductivity), but a slight and variable depletion of non-structural carbohydrate reserves preceding dieback. Five- and six-year-old trees of U. minor (planted in the field with supplementary watering) belonging to clones of contrasted susceptibility to DED exhibited a different physiological response to inoculation with O. novo-ulmi. Stem hydraulic conductivity, leaf water potential and photosynthetic CO2 uptake decreased significantly relative to control trees inoculated with water only in DED susceptible clones. This is consistent with the “more defensive” chemical profile observed in resistant clones, i.e. with higher levels of saturated hydrocarbons (suberin and fatty acids) and phenolic compounds than in susceptible clones. These compounds could restrict the spread of O. novo-ulmi and contribute to preserving the near-normal physiological function of resistant trees when exposed to the pathogen. These results evidence common physiological responses of U. minor to flooding, drought and pathogen infection leading to xylem water disruption, leaf water stress and reduced net carbon gain. Still, seedlings of U. minor develop various mechanisms of acclimation to abiotic stresses that can play a role in surviving moderate periods of flood and drought. The chemical profile appears to be an important factor for the resistance of some genotypes of U. minor to DED. How abiotic stresses such as flooding and drought affect the capacity of resistant U. minor clones to face O. novo-ulmi is a key question that must be contemplated in future research.

Uso do protocolo no serviço de psicologia e avaliação psicodinâmica em candidatos à cirurgia bariátrica

A clínica da Obesidade Mórbida e a Cirurgia Bariátrica exige estudos e acompanhamentos do paciente. Os benefícios e riscos do emagrecimento por tratamento cirúrgico devem servir como ponto de alerta aos profissionais da saúde. O uso do questionário no serviço de psicologia é norteado pela escuta psicológica. Objetivos: 1) Descrever o perfil sócio-demográfico candidatos à cirurgia bariátrica. 2) Analisar a percepção dos pacientes sobre características de personalidade associadas à obesidade e transtornos alimentares. 3) Descrever os conteúdos psicodinâmicos da narrativa do sujeito e avaliar o sistema tensional inconsciente de dois pacientes por meio do Teste das Relações Objetais de Phillipson (TRO). Método: O delineamento metodológico com análise de dados pelo método epidemiológico e estudo de caso clínico, orientação psicanalítica. Na primeira etapa foram consultados 300 questionários do serviço de psicologia e na segunda dois pacientes com ganho de peso após 24 meses. São pacientes que procuraram tratamento em clínica especializada, em uma metrópole do sudeste brasileiro, sob consentimento pós-informado. Os questionários foram preenchidos por 227 mulheres e 73 homens; com média de idade igual a 36 anos; escolaridade ensino médio e superior, 53%; maioria casados; IMC entre grave e super mórbido (94,3%). Técnicas cirúrgicas indicadas Capella Bypass e Fobi-Capella (67%). Resultados: características psicológicas referidas pelos pacientes, a ansiedade apontou em 93,7% das respostas, seguidas por impulsividade, depressão, tolerância à frustração, baixa auto-estima, resolvedor de problemas dos outros (mais de 50%). No histórico familiar da obesidade está em mais de 70% depressão e uso do álcool em 30%; realização de psicoterapia (30%) e medicamentos para depressão e ansiedade (10%). Na segunda etapa, foi realizado o diagnóstico psicodinâmico, por meio do Teste das Relações Objetais de Phillipson com duas pacientes, cuja análise indicou necessidade de psicoterapia psicanalítica, pois tinham fixações na posição esquizoparanóide e apresentavam dificuldade em lidar com perdas e baixa motivação para mudança e insigth. Conclusões: Com a aplicação do questionário e o registro das observações empíricas, este questionário de entrevista semidirigida preenche condições de melhor acessar e avaliar os conteúdos revelados pelos pacientes. As contradições entre as respostas e o discurso, no contato individual com o psicólogo, apontam a necessidade de investimento no preparo do paciente para a cirurgia e mais acentuadamente o acompanhamento psicológico no primeiro ano do pós-operatório. Há um pensamento mágico a ser trabalhado durante a aplicação do questionário sobre as crenças frente à cirurgia e o emagrecimento e assim convocar o paciente a ocupar o lugar do sujeito implicado em seu processo pré e pós-operatório. O TRO contribuiu na compreensão do diagnóstico psicodinâmico de pacientes com ganho de peso após cirurgia e reforçou a necessidade de maior investimento no pré-operatório.(AU)

Uso do protocolo no serviço de psicologia e avaliação psicodinâmica em candidatos à cirurgia bariátrica

A clínica da Obesidade Mórbida e a Cirurgia Bariátrica exige estudos e acompanhamentos do paciente. Os benefícios e riscos do emagrecimento por tratamento cirúrgico devem servir como ponto de alerta aos profissionais da saúde. O uso do questionário no serviço de psicologia é norteado pela escuta psicológica. Objetivos: 1) Descrever o perfil sócio-demográfico candidatos à cirurgia bariátrica. 2) Analisar a percepção dos pacientes sobre características de personalidade associadas à obesidade e transtornos alimentares. 3) Descrever os conteúdos psicodinâmicos da narrativa do sujeito e avaliar o sistema tensional inconsciente de dois pacientes por meio do Teste das Relações Objetais de Phillipson (TRO). Método: O delineamento metodológico com análise de dados pelo método epidemiológico e estudo de caso clínico, orientação psicanalítica. Na primeira etapa foram consultados 300 questionários do serviço de psicologia e na segunda dois pacientes com ganho de peso após 24 meses. São pacientes que procuraram tratamento em clínica especializada, em uma metrópole do sudeste brasileiro, sob consentimento pós-informado. Os questionários foram preenchidos por 227 mulheres e 73 homens; com média de idade igual a 36 anos; escolaridade ensino médio e superior, 53%; maioria casados; IMC entre grave e super mórbido (94,3%). Técnicas cirúrgicas indicadas Capella Bypass e Fobi-Capella (67%). Resultados: características psicológicas referidas pelos pacientes, a ansiedade apontou em 93,7% das respostas, seguidas por impulsividade, depressão, tolerância à frustração, baixa auto-estima, resolvedor de problemas dos outros (mais de 50%). No histórico familiar da obesidade está em mais de 70% depressão e uso do álcool em 30%; realização de psicoterapia (30%) e medicamentos para depressão e ansiedade (10%). Na segunda etapa, foi realizado o diagnóstico psicodinâmico, por meio do Teste das Relações Objetais de Phillipson com duas pacientes, cuja análise indicou necessidade de psicoterapia psicanalítica, pois tinham fixações na posição esquizoparanóide e apresentavam dificuldade em lidar com perdas e baixa motivação para mudança e insigth. Conclusões: Com a aplicação do questionário e o registro das observações empíricas, este questionário de entrevista semidirigida preenche condições de melhor acessar e avaliar os conteúdos revelados pelos pacientes. As contradições entre as respostas e o discurso, no contato individual com o psicólogo, apontam a necessidade de investimento no preparo do paciente para a cirurgia e mais acentuadamente o acompanhamento psicológico no primeiro ano do pós-operatório. Há um pensamento mágico a ser trabalhado durante a aplicação do questionário sobre as crenças frente à cirurgia e o emagrecimento e assim convocar o paciente a ocupar o lugar do sujeito implicado em seu processo pré e pós-operatório. O TRO contribuiu na compreensão do diagnóstico psicodinâmico de pacientes com ganho de peso após cirurgia e reforçou a necessidade de maior investimento no pré-operatório.(AU)

Transcription initiation is controlled by three core promoter elements in the hgl5 gene of the protozoan parasite Entamoeba histolytica

Entamoeba histolytica is a single cell eukaryote that is the etiologic agent of amoebic colitis. Core promoter elements of E. histolytica protein encoding genes include a TATA-like sequence (GTATTTAAAG/C) at −30, a novel element designated GAAC (GAACT) that has a variable location between TATA and the site of transcription initiation, and a putative initiator (Inr) element (AAAAATTCA) overlying the site of transcription initiation. The presence of three separate conserved sequences in a eukaryotic core promoter is unprecedented and prompted examination of their roles in regulating transcription initiation. Alterations of all three regions in the hgl5 gene decreased reporter gene activity with the greatest effect seen by mutation of the GAAC element. Positional analysis of the TATA box demonstrated that transcription initiated consistently 30–31 bases downstream of the TATA region. Mutation of either the TATA or GAAC elements resulted in the appearance of new transcription start sites upstream of +1 in the promoter of the hgl5 gene. Mutation of the Inr element resulted in no change in the site of transcription initiation; however, in the presence of a mutated TATA and GAAC regions, the Inr element controlled the site of transcription initiation. We conclude that all three elements play a role in determining the site of transcription initiation. The variable position of the GAAC element relative to the site of transcription initiation, and the multiple transcription initiations that resulted from its mutation, indicate that the GAAC element has an important and apparently novel role in transcriptional control in E. histolytica.

Neural mechanisms underlying binocular fusion and stereopsis: Position vs. phase

The visual system utilizes binocular disparity to discriminate the relative depth of objects in space. Since the striate cortex is the first site along the central visual pathways at which signals from the left and right eyes converge onto a single neuron, encoding of binocular disparity is thought to begin in this region. There are two possible mechanisms for encoding binocular disparity through simple cells in the striate cortex: a difference in receptive field (RF) position between the two eyes (RF position disparity) and a difference in RF profile between the two eyes (RF phase disparity). Although there have been studies supporting each of the two encoding mechanisms, both mechanisms have not been examined in a single study. Therefore, the relative roles of the two mechanisms have not been determined. To address this issue, we have mapped left and right eye RFs of simple cells in the cat’s striate cortex using binary m-sequence noise, and then we have estimated RF position and phase disparities. We find that RF position disparities are generally limited to small values that are not sufficient to encode large binocular disparities. In contrast, RF phase disparities cover a wide range of binocular disparities and exhibit dependencies on orientation and spatial frequency in a manner expected for a mechanism that encodes binocular disparity. These results indicate that binocular disparity is mainly encoded through RF phase disparity. However, RF position disparity may play a significant role for cells with high spatial frequency selectivity, which are constrained to small RF phase disparities.

Sorbitol-6-Phosphate Dehydrogenase Expression in Transgenic Tobacco1 : High Amounts of Sorbitol Lead to Necrotic Lesions

We analyzed transgenic tobacco (Nicotiana tabacum L.) expressing Stpd1, a cDNA encoding sorbitol-6-phosphate dehydrogenase from apple, under the control of a cauliflower mosaic virus 35S promoter. In 125 independent transformants variable amounts of sorbitol ranging from 0.2 to 130 μmol g−1 fresh weight were found. Plants that accumulated up to 2 to 3 μmol g−1 fresh weight sorbitol were phenotypically normal, with successively slower growth as sorbitol amounts increased. Plants accumulating sorbitol at 3 to 5 μmol g−1 fresh weight occasionally showed regions in which chlorophyll was partially lost, but at higher sorbitol amounts young leaves of all plants lost chlorophyll in irregular spots that developed into necrotic lesions. When sorbitol exceeded 15 to 20 μmol g−1 fresh weight, plants were infertile, and at even higher sorbitol concentrations the primary regenerants were incapable of forming roots in culture or soil. In mature plants sorbitol amounts varied with age, leaf position, and growth conditions. The appearance of lesions was correlated with high sorbitol, glucose, fructose, and starch, and low myo-inositol. Supplementing myo-inositol in seedlings and young plants prevented lesion formation. Hyperaccumulation of sorbitol, which interferes with inositol biosynthesis, seems to lead to osmotic imbalance, possibly acting as a signal affecting carbohydrate allocation and transport.

Generation of a high-titer retroviral vector capable of expressing high levels of the human beta-globin gene.

Retrovirus-mediated gene transfer into hematopoietic cells may provide a means of treating both inherited and acquired diseases involving hematopoietic cells. Implementation of this approach for disorders resulting from mutations affecting the beta-globin gene (e.g., beta-thalassemia and sickle cell anemia), however, has been hampered by the inability to generate recombinant viruses able to efficiently and faithfully transmit the necessary sequences for appropriate gene expression. We have addressed this problem by carefully examining the interactions between retroviral and beta-globin gene sequences which affect vector transmission, stability, and expression. First, we examined the transmission properties of a large number of different recombinant proviral genomes which vary both in the precise nature of vector, beta-globin structural gene, and locus control region (LCR) core sequences incorporated and in the placement and orientation of those sequences. Through this analysis, we identified one specific vector, termed M beta 6L, which carries both the human beta-globin gene and core elements HS2, HS3, and HS4 from the LCR and faithfully transmits recombinant proviral sequences to cells with titers greater than 10(6) per ml. Populations of murine erythroleukemia (MEL) cells transduced by this virus expressed levels of human beta-globin transcript which, on a per gene copy basis, were 78% of the levels detected in an MEL-derived cell line, Hu11, which carries human chromosome 11, the site of the beta-globin locus. Analysis of individual transduced MEL cell clones, however, indicated that, while expression was detected in every clone tested (n = 17), the levels of human beta-globin treatment varied between 4% and 146% of the levels in Hu11. This clonal variation in expression levels suggests that small beta-globin LCR sequences may not provide for as strict chromosomal position-independent expression of beta-globin as previously suspected, at least in the context of retrovirus-mediated gene transfer.

Narratives and counter-narratives on "marriage of convenience" : conjugality and (il)legality in portuguese migration policies and in couples’ experiences

Tese de doutoramento, Sociologia (Sociologia da Família, Juventude e das Relações do Género), Universidade de Lisboa, Instituto de Ciências Sociais, 2016

Impacts fonctionnels du traumatisme craniocérébral léger sur la vision et l'équilibre postural chez l'adulte

Le traumatisme craniocérébral léger (TCCL) a des effets complexes sur plusieurs fonctions cérébrales, dont l’évaluation et le suivi peuvent être difficiles. Les problèmes visuels et les troubles de l’équilibre font partie des plaintes fréquemment rencontrées après un TCCL. En outre, ces problèmes peuvent continuer à affecter les personnes ayant eu un TCCL longtemps après la phase aiguë du traumatisme. Cependant, les évaluations cliniques conventionnelles de la vision et de l’équilibre ne permettent pas, la plupart du temps, d’objectiver ces symptômes, surtout lorsqu’ils s’installent durablement. De plus, il n’existe pas, à notre connaissance, d’étude longitudinale ayant étudié les déficits visuels perceptifs, en tant que tels, ni les troubles de l’équilibre secondaires à un TCCL, chez l’adulte. L’objectif de ce projet était donc de déterminer la nature et la durée des effets d’un tel traumatisme sur la perception visuelle et sur la stabilité posturale, en évaluant des adultes TCCL et contrôles sur une période d’un an. Les mêmes sujets, exactement, ont participé aux deux expériences, qui ont été menées les mêmes jours pour chacun des sujets. L’impact du TCCL sur la perception visuelle de réseaux sinusoïdaux définis par des attributs de premier et de second ordre a d’abord été étudié. Quinze adultes diagnostiqués TCCL ont été évalués 15 jours, 3 mois et 12 mois après leur traumatisme. Quinze adultes contrôles appariés ont été évalués à des périodes identiques. Des temps de réaction (TR) de détection de clignotement et de discrimination de direction de mouvement ont été mesurés. Les niveaux de contraste des stimuli de premier et de second ordre ont été ajustés pour qu’ils aient une visibilité comparable, et les moyennes, médianes, écarts-types (ET) et écarts interquartiles (EIQ) des TR correspondant aux bonnes réponses ont été calculés. Le niveau de symptômes a également été évalué pour le comparer aux données de TR. De façon générale, les TR des TCCL étaient plus longs et plus variables (plus grands ET et EIQ) que ceux des contrôles. De plus, les TR des TCCL étaient plus courts pour les stimuli de premier ordre que pour ceux de second ordre, et plus variables pour les stimuli de premier ordre que pour ceux de second ordre, dans la condition de discrimination de mouvement. Ces observations se sont répétées au cours des trois sessions. Le niveau de symptômes des TCCL était supérieur à celui des participants contrôles, et malgré une amélioration, cet écart est resté significatif sur la période d’un an qui a suivi le traumatisme. La seconde expérience, elle, était destinée à évaluer l’impact du TCCL sur le contrôle postural. Pour cela, nous avons mesuré l’amplitude d’oscillation posturale dans l’axe antéropostérieur et l’instabilité posturale (au moyen de la vitesse quadratique moyenne (VQM) des oscillations posturales) en position debout, les pieds joints, sur une surface ferme, dans cinq conditions différentes : les yeux fermés, et dans un tunnel virtuel tridimensionnel soit statique, soit oscillant de façon sinusoïdale dans la direction antéropostérieure à trois vitesses différentes. Des mesures d’équilibre dérivées de tests cliniques, le Bruininks-Oseretsky Test of Motor Proficiency 2nd edition (BOT-2) et le Balance Error Scoring System (BESS) ont également été utilisées. Les participants diagnostiqués TCCL présentaient une plus grande instabilité posturale (une plus grande VQM des oscillations posturales) que les participants contrôles 2 semaines et 3 mois après le traumatisme, toutes conditions confondues. Ces troubles de l’équilibre secondaires au TCCL n’étaient plus présents un an après le traumatisme. Ces résultats suggèrent également que les déficits affectant les processus d’intégration visuelle mis en évidence dans la première expérience ont pu contribuer aux troubles de l’équilibre secondaires au TCCL. L’amplitude d’oscillation posturale dans l’axe antéropostérieur de même que les mesures dérivées des tests cliniques d’évaluation de l’équilibre (BOT-2 et BESS) ne se sont pas révélées être des mesures sensibles pour quantifier le déficit postural chez les sujets TCCL. L’association des mesures de TR à la perception des propriétés spécifiques des stimuli s’est révélée être à la fois une méthode de mesure particulièrement sensible aux anomalies visuomotrices secondaires à un TCCL, et un outil précis d’investigation des mécanismes sous-jacents à ces anomalies qui surviennent lorsque le cerveau est exposé à un traumatisme léger. De la même façon, les mesures d’instabilité posturale se sont révélées suffisamment sensibles pour permettre de mesurer les troubles de l’équilibre secondaires à un TCCL. Ainsi, le développement de tests de dépistage basés sur ces résultats et destinés à l’évaluation du TCCL dès ses premières étapes apparaît particulièrement intéressant. Il semble également primordial d’examiner les relations entre de tels déficits et la réalisation d’activités de la vie quotidienne, telles que les activités scolaires, professionnelles ou sportives, pour déterminer les impacts fonctionnels que peuvent avoir ces troubles des fonctions visuomotrice et du contrôle de l’équilibre.