922 resultados para High-level Design Specification
Resumo:
El campo de estudio relacionado con los laboratorios remotos en el ámbito educativo de las ciencias y la ingeniería está sufriendo una notable expansión ante la necesidad de adaptar los procesos de aprendizaje en dichas áreas a las características y posibilidades de la formación online. Muchos de los recursos educativos basados en esta tecnología, existentes en la actualidad, presentan ciertas limitaciones que impiden alcanzar las competencias que se deben adquirir en los laboratorios de ingeniería. Estas limitaciones están relacionadas con diferentes aspectos de carácter técnico y formativo. A nivel técnico las limitaciones principales se centran en el grado de versatilidad que son capaces de proporcionar comparado con el que se dispone en un laboratorio tradicional y en el modo de interacción del usuario, que provoca que el estudiante no distinga claramente si está realizando acciones sobre sistemas reales o simulaciones. A nivel formativo las limitaciones detectadas son relevantes para poder alcanzar un aprendizaje significativo. En concreto están relacionadas principalmente con un escaso sentimiento de inmersión, una reducida sensación de realismo respecto a las operaciones que se realizan o la limitada posibilidad de realizar actividades de forma colaborativa. La aparición de nuevas tecnologías basadas en entornos inmersivos, unida a los avances producidos relacionados con el aumento de la capacidad gráfica de los ordenadores y del ancho de banda de acceso a Internet, han hecho factible que las limitaciones comentadas anteriormente puedan ser superadas gracias al desarrollo de nuevos recursos de aprendizaje surgidos de la fusión de laboratorios remotos y mundos virtuales 3D. Esta tesis doctoral aborda un trabajo de investigación centrado en proponer un modelo de plataformas experimentales, basado en la fusión de las dos tecnologías mencionadas, que permita generar recursos educativos online que faciliten la adquisición de competencias prácticas similares a las que se consiguen en un laboratorio tradicional vinculado a la enseñanza de la electrónica. El campo de aplicación en el que se ha focalizado el trabajo realizado se ha centrado en el área de la electrónica aunque los resultados de la investigación realizada se podrían adaptar fácilmente a otras disciplinas de la ingeniería. Fruto del trabajo realizado en esta tesis es el desarrollo de la plataforma eLab3D, basada en el modelo de plataformas experimentales propuesto, y la realización de dos estudios empíricos llevados a cabo con estudiantes de grado en ingeniería, muy demandados por la comunidad investigadora. Por un lado, la plataforma eLab3D, que permite llevar a cabo de forma remota actividades prácticas relacionadas con el diseño, montaje y prueba de circuitos electrónicos analógicos, aporta como novedad un dispositivo hardware basado en un sistema de conmutación distribuido. Dicho sistema proporciona un nivel de versatilidad muy elevado, a nivel de configuración de circuitos y selección de puntos de medida, que hace posible la realización de acciones similares a las que se llevan a cabo en los laboratorios presenciales. Por otra parte, los estudios empíricos realizados, que comparaban la eficacia educativa de una metodología de aprendizaje online, basada en el uso de la plataforma eLab3D, con la conseguida siguiendo una metodología clásica en los laboratorios tradicionales, mostraron que no se detectaron diferencias significativas en el grado de adquisición de los resultados de aprendizaje entre los estudiantes que utilizaron la plataforma eLab3D y los que asistieron a los laboratorios presenciales. Por último, hay que destacar dos aspectos relevantes relacionados directamente con esta tesis. En primer lugar, los resultados obtenidos en las experiencias educativas llevadas a cabo junto a valoraciones obtenidas por el profesorado que ha colaborado en las mismas han sido decisivos para que la plataforma eLab3D se haya integrado como recurso complementario de aprendizaje en titulaciones de grado de ingeniería de la Universidad Politécnica de Madrid. En segundo lugar, el modelo de plataformas experimentales que se ha propuesto en esta tesis, analizado por investigadores vinculados a proyectos en el ámbito de la fusión nuclear, ha sido tomado como referencia para generar nuevas herramientas de formación en dicho campo. ABSTRACT The field of study of remote laboratories in sciences and engineering educational disciplines is undergoing a remarkable expansion given the need to adapt the learning processes in the aforementioned areas to the characteristics and possibilities of online education. Several of the current educational resources based on this technology have certain limitations that prevent from reaching the required competencies in engineering laboratories. These limitations are related to different aspects of technical and educational nature. At the technical level, they are centered on the degree of versatility they are able to provide compared to a traditional laboratory and in the way the user interacts with them, which causes the student to not clearly distinguish if actions are being performed over real systems or over simulations. At the educational level, the detected limitations are relevant in order to reach a meaningful learning. In particular, they are mainly related to a scarce immersion feeling, a reduced realism sense regarding the operations performed or the limited possibility to carry out activities in a collaborative way. The appearance of new technologies based on immersive environments, together with the advances in graphical computer capabilities and Internet bandwidth access, have made the previous limitations feasible to be overcome thanks to the development of new learning resources that arise from merging remote laboratories and 3D virtual worlds. This PhD thesis tackles a research work focused on the proposal of an experimental platform model, based on the fusion of both mentioned technologies, which allows for generating online educational resources that facilitate the acquisition of practical competencies similar to those obtained in a traditional electronics laboratory. The application field, in which this work is focused, is electronics, although the research results could be easily adapted to other engineering disciplines. A result of this work is the development of eLab3D platform, based on the experimental platform model proposed, and the realization of two empirical studies with undergraduate students, highly demanded by research community. On one side, eLab3D platform, which allows to accomplish remote practical activities related to the design, assembling and test of analog electronic circuits, provides, as an original contribution, a hardware device based on a distributed switching system. This system offers a high level of versatility, both at the circuit configuration level and at the selection of measurement points, which allows for doing similar actions to those conducted in hands-on laboratories. On the other side, the empirical studies carried out, which compare the educational efficiency of an online learning methodology based on the use of eLab3D platform with that obtained following a classical methodology in traditional laboratories, shows that no significant differences in the acquired degree of learning outcomes among the students that used eLab3D platform and those that attended hands-on laboratories were detected. Finally, it is important to highlight two relevant aspects directly related with this thesis work. First of all, the results obtained in the educational experiences conducted, along with the assessment from the faculty that has collaborated in them, have been decisive to integrate eLab3D platform as a supplementary learning resource in engineering degrees at Universidad Politecnica de Madrid. Secondly, the experimental platform model originally proposed in this thesis, which has been analysed by nuclear fusion researchers, has been taken as a reference to generate new educational tools in that field.
Resumo:
Emotion is generally argued to be an influence on the behavior of life systems, largely concerning flexibility and adaptivity. The way in which life systems acts in response to a particular situations of the environment, has revealed the decisive and crucial importance of this feature in the success of behaviors. And this source of inspiration has influenced the way of thinking artificial systems. During the last decades, artificial systems have undergone such an evolution that each day more are integrated in our daily life. They have become greater in complexity, and the subsequent effects are related to an increased demand of systems that ensure resilience, robustness, availability, security or safety among others. All of them questions that raise quite a fundamental challenges in control design. This thesis has been developed under the framework of the Autonomous System project, a.k.a the ASys-Project. Short-term objectives of immediate application are focused on to design improved systems, and the approaching of intelligence in control strategies. Besides this, long-term objectives underlying ASys-Project concentrate on high order capabilities such as cognition, awareness and autonomy. This thesis is placed within the general fields of Engineery and Emotion science, and provides a theoretical foundation for engineering and designing computational emotion for artificial systems. The starting question that has grounded this thesis aims the problem of emotion--based autonomy. And how to feedback systems with valuable meaning has conformed the general objective. Both the starting question and the general objective, have underlaid the study of emotion, the influence on systems behavior, the key foundations that justify this feature in life systems, how emotion is integrated within the normal operation, and how this entire problem of emotion can be explained in artificial systems. By assuming essential differences concerning structure, purpose and operation between life and artificial systems, the essential motivation has been the exploration of what emotion solves in nature to afterwards analyze analogies for man--made systems. This work provides a reference model in which a collection of entities, relationships, models, functions and informational artifacts, are all interacting to provide the system with non-explicit knowledge under the form of emotion-like relevances. This solution aims to provide a reference model under which to design solutions for emotional operation, but related to the real needs of artificial systems. The proposal consists of a multi-purpose architecture that implement two broad modules in order to attend: (a) the range of processes related to the environment affectation, and (b) the range or processes related to the emotion perception-like and the higher levels of reasoning. This has required an intense and critical analysis beyond the state of the art around the most relevant theories of emotion and technical systems, in order to obtain the required support for those foundations that sustain each model. The problem has been interpreted and is described on the basis of AGSys, an agent assumed with the minimum rationality as to provide the capability to perform emotional assessment. AGSys is a conceptualization of a Model-based Cognitive agent that embodies an inner agent ESys, the responsible of performing the emotional operation inside of AGSys. The solution consists of multiple computational modules working federated, and aimed at conforming a mutual feedback loop between AGSys and ESys. Throughout this solution, the environment and the effects that might influence over the system are described as different problems. While AGSys operates as a common system within the external environment, ESys is designed to operate within a conceptualized inner environment. And this inner environment is built on the basis of those relevances that might occur inside of AGSys in the interaction with the external environment. This allows for a high-quality separate reasoning concerning mission goals defined in AGSys, and emotional goals defined in ESys. This way, it is provided a possible path for high-level reasoning under the influence of goals congruence. High-level reasoning model uses knowledge about emotional goals stability, letting this way new directions in which mission goals might be assessed under the situational state of this stability. This high-level reasoning is grounded by the work of MEP, a model of emotion perception that is thought as an analogy of a well-known theory in emotion science. The work of this model is described under the operation of a recursive-like process labeled as R-Loop, together with a system of emotional goals that are assumed as individual agents. This way, AGSys integrates knowledge that concerns the relation between a perceived object, and the effect which this perception induces on the situational state of the emotional goals. This knowledge enables a high-order system of information that provides the sustain for a high-level reasoning. The extent to which this reasoning might be approached is just delineated and assumed as future work. This thesis has been studied beyond a long range of fields of knowledge. This knowledge can be structured into two main objectives: (a) the fields of psychology, cognitive science, neurology and biological sciences in order to obtain understanding concerning the problem of the emotional phenomena, and (b) a large amount of computer science branches such as Autonomic Computing (AC), Self-adaptive software, Self-X systems, Model Integrated Computing (MIC) or the paradigm of models@runtime among others, in order to obtain knowledge about tools for designing each part of the solution. The final approach has been mainly performed on the basis of the entire acquired knowledge, and described under the fields of Artificial Intelligence, Model-Based Systems (MBS), and additional mathematical formalizations to provide punctual understanding in those cases that it has been required. This approach describes a reference model to feedback systems with valuable meaning, allowing for reasoning with regard to (a) the relationship between the environment and the relevance of the effects on the system, and (b) dynamical evaluations concerning the inner situational state of the system as a result of those effects. And this reasoning provides a framework of distinguishable states of AGSys derived from its own circumstances, that can be assumed as artificial emotion.
Resumo:
Uno de los mayores retos para la comunidad científica es conseguir que las máquinas posean en un futuro la capacidad del sistema visual y cognitivo humanos, de forma que, por ejemplo, en entornos de video vigilancia, puedan llegar a proporcionar de manera automática una descripción fiable de lo que está ocurriendo en la escena. En la presente tesis, mediante la propuesta de un marco de trabajo de referencia, se discuten y plantean los pasos necesarios para el desarrollo de sistemas más inteligentes capaces de extraer y analizar, a diferentes niveles de abstracción y mediante distintos módulos de procesamiento independientes, la información necesaria para comprender qué está sucediendo en un conjunto amplio de escenarios de distinta naturaleza. Se parte de un análisis de requisitos y se identifican los retos para este tipo de sistemas en la actualidad, lo que constituye en sí mismo los objetivos de esta tesis, contribuyendo así a un modelo de datos basado en el conocimiento que permitirá analizar distintas situaciones en las que personas y vehículos son los actores principales, dejando no obstante la puerta abierta a la adaptación a otros dominios. Así mismo, se estudian los distintos procesos que se pueden lanzar a nivel interno así como la necesidad de integrar mecanismos de realimentación a distintos niveles que permitan al sistema adaptarse mejor a cambios en el entorno. Como resultado, se propone un marco de referencia jerárquico que integra las capacidades de percepción, interpretación y aprendizaje para superar los retos identificados en este ámbito; y así poder desarrollar sistemas de vigilancia más robustos, flexibles e inteligentes, capaces de operar en una variedad de entornos. Resultados experimentales ejecutados sobre distintas muestras de datos (secuencias de vídeo principalmente) demuestran la efectividad del marco de trabajo propuesto respecto a otros propuestos en el pasado. Un primer caso de estudio, permite demostrar la creación de un sistema de monitorización de entornos de parking en exteriores para la detección de vehículos y el análisis de plazas libres de aparcamiento. Un segundo caso de estudio, permite demostrar la flexibilidad del marco de referencia propuesto para adaptarse a los requisitos de un entorno de vigilancia completamente distinto, como es un hogar inteligente donde el análisis automático de actividades de la vida cotidiana centra la atención del estudio. ABSTRACT One of the most ambitious objectives for the Computer Vision and Pattern Recognition research community is that machines can achieve similar capacities to the human's visual and cognitive system, and thus provide a trustworthy description of what is happening in the scene under surveillance. Thus, a number of well-established scenario understanding architectural frameworks to develop applications working on a variety of environments can be found in the literature. In this Thesis, a highly descriptive methodology for the development of scene understanding applications is presented. It consists of a set of formal guidelines to let machines extract and analyse, at different levels of abstraction and by means of independent processing modules that interact with each other, the necessary information to understand a broad set of different real World surveillance scenarios. Taking into account the challenges that working at both low and high levels offer, we contribute with a highly descriptive knowledge-based data model for the analysis of different situations in which people and vehicles are the main actors, leaving the door open for the development of interesting applications in diverse smart domains. Recommendations to let systems achieve high-level behaviour understanding will be also provided. Furthermore, feedback mechanisms are proposed to be integrated in order to let any system to understand better the environment and the logical context around, reducing thus the uncertainty and noise, and increasing its robustness and precision in front of low-level or high-level errors. As a result, a hierarchical cognitive architecture of reference which integrates the necessary perception, interpretation, attention and learning capabilities to overcome main challenges identified in this area of research is proposed; thus allowing to develop more robust, flexible and smart surveillance systems to cope with the different requirements of a variety of environments. Once crucial issues that should be treated explicitly in the design of this kind of systems have been formulated and discussed, experimental results shows the effectiveness of the proposed framework compared with other proposed in the past. Two case studies were implemented to test the capabilities of the framework. The first case study presents how the proposed framework can be used to create intelligent parking monitoring systems. The second case study demonstrates the flexibility of the system to cope with the requirements of a completely different environment, a smart home where activities of daily living are performed. Finally, general conclusions and future work lines to further enhancing the capabilities of the proposed framework are presented.
Resumo:
En el futuro, la gestión del tráfico aéreo (ATM, del inglés air traffic management) requerirá un cambio de paradigma, de la gestión principalmente táctica de hoy, a las denominadas operaciones basadas en trayectoria. Un incremento en el nivel de automatización liberará al personal de ATM —controladores, tripulación, etc.— de muchas de las tareas que realizan hoy. Las personas seguirán siendo el elemento central en la gestión del tráfico aéreo del futuro, pero lo serán mediante la gestión y toma de decisiones. Se espera que estas dos mejoras traigan un incremento en la eficiencia de la gestión del tráfico aéreo que permita hacer frente al incremento previsto en la demanda de transporte aéreo. Para aplicar el concepto de operaciones basadas en trayectoria, el usuario del espacio aéreo (la aerolínea, piloto, u operador) y el proveedor del servicio de navegación aérea deben negociar las trayectorias mediante un proceso de toma de decisiones colaborativo. En esta negociación, es necesaria una forma adecuada de compartir dichas trayectorias. Compartir la trayectoria completa requeriría un gran ancho de banda, y la trayectoria compartida podría invalidarse si cambiase la predicción meteorológica. En su lugar, podría compartirse una descripción de la trayectoria independiente de las condiciones meteorológicas, de manera que la trayectoria real se pudiese calcular a partir de dicha descripción. Esta descripción de la trayectoria debería ser fácil de procesar usando un programa de ordenador —ya que parte del proceso de toma de decisiones estará automatizado—, pero también fácil de entender para un operador humano —que será el que supervise el proceso y tome las decisiones oportunas—. Esta tesis presenta una serie de lenguajes formales que pueden usarse para este propósito. Estos lenguajes proporcionan los medios para describir trayectorias de aviones durante todas las fases de vuelo, desde la maniobra de push-back (remolcado hasta la calle de rodaje), hasta la llegada a la terminal del aeropuerto de destino. También permiten describir trayectorias tanto de aeronaves tripuladas como no tripuladas, incluyendo aviones de ala fija y cuadricópteros. Algunos de estos lenguajes están estrechamente relacionados entre sí, y organizados en una jerarquía. Uno de los lenguajes fundamentales de esta jerarquía, llamado aircraft intent description language (AIDL), ya había sido desarrollado con anterioridad a esta tesis. Este lenguaje fue derivado de las ecuaciones del movimiento de los aviones de ala fija, y puede utilizarse para describir sin ambigüedad trayectorias de este tipo de aeronaves. Una variante de este lenguaje, denominada quadrotor AIDL (QR-AIDL), ha sido desarrollada en esta tesis para permitir describir trayectorias de cuadricópteros con el mismo nivel de detalle. Seguidamente, otro lenguaje, denominado intent composite description language (ICDL), se apoya en los dos lenguajes anteriores, ofreciendo más flexibilidad para describir algunas partes de la trayectoria y dejar otras sin especificar. El ICDL se usa para proporcionar descripciones genéricas de maniobras comunes, que después se particularizan y combinan para formar descripciones complejas de un vuelo. Otro lenguaje puede construirse a partir del ICDL, denominado flight intent description language (FIDL). El FIDL especifica requisitos de alto nivel sobre las trayectorias —incluyendo restricciones y objetivos—, pero puede utilizar características del ICDL para proporcionar niveles de detalle arbitrarios en las distintas partes de un vuelo. Tanto el ICDL como el FIDL han sido desarrollados en colaboración con Boeing Research & Technology Europe (BR&TE). También se ha desarrollado un lenguaje para definir misiones en las que interactúan varias aeronaves, el mission intent description language (MIDL). Este lenguaje se basa en el FIDL y mantiene todo su poder expresivo, a la vez que proporciona nuevas semánticas para describir tareas, restricciones y objetivos relacionados con la misión. En ATM, los movimientos de un avión en la superficie de aeropuerto también tienen que ser monitorizados y gestionados. Otro lenguaje formal ha sido diseñado con este propósito, llamado surface movement description language (SMDL). Este lenguaje no pertenece a la jerarquía de lenguajes descrita en el párrafo anterior, y se basa en las clearances (autorizaciones del controlador) utilizadas durante las operaciones en superficie de aeropuerto. También proporciona medios para expresar incertidumbre y posibilidad de cambios en las distintas partes de la trayectoria. Finalmente, esta tesis explora las aplicaciones de estos lenguajes a la predicción de trayectorias y a la planificación de misiones. El concepto de trajectory language processing engine (TLPE) se usa en ambas aplicaciones. Un TLPE es una función de ATM cuya principal entrada y salida se expresan en cualquiera de los lenguajes incluidos en la jerarquía descrita en esta tesis. El proceso de predicción de trayectorias puede definirse como una combinación de TLPEs, cada uno de los cuales realiza una pequeña sub-tarea. Se le ha dado especial importancia a uno de estos TLPEs, que se encarga de generar el perfil horizontal, vertical y de configuración de la trayectoria. En particular, esta tesis presenta un método novedoso para la generación del perfil vertical. El proceso de planificar una misión también se puede ver como un TLPE donde la entrada se expresa en MIDL y la salida consiste en cierto número de trayectorias —una por cada aeronave disponible— descritas utilizando FIDL. Se ha formulado este problema utilizando programación entera mixta. Además, dado que encontrar caminos óptimos entre distintos puntos es un problema fundamental en la planificación de misiones, también se propone un algoritmo de búsqueda de caminos. Este algoritmo permite calcular rápidamente caminos cuasi-óptimos que esquivan todos los obstáculos en un entorno urbano. Los diferentes lenguajes formales definidos en esta tesis pueden utilizarse como una especificación estándar para la difusión de información entre distintos actores de la gestión del tráfico aéreo. En conjunto, estos lenguajes permiten describir trayectorias con el nivel de detalle necesario en cada aplicación, y se pueden utilizar para aumentar el nivel de automatización explotando esta información utilizando sistemas de soporte a la toma de decisiones. La aplicación de estos lenguajes a algunas funciones básicas de estos sistemas, como la predicción de trayectorias, han sido analizadas. ABSTRACT Future air traffic management (ATM) will require a paradigm shift from today’s mainly tactical ATM to trajectory-based operations (TBOs). An increase in the level of automation will also relieve humans —air traffic control officers (ATCOs), flight crew, etc.— from many of the tasks they perform today. Humans will still be central in this future ATM, as decision-makers and managers. These two improvements (TBOs and increased automation) are expected to provide the increase in ATM performance that will allow coping with the expected increase in air transport demand. Under TBOs, trajectories are negotiated between the airspace user (an airline, pilot, or operator) and the air navigation service provider (ANSP) using a collaborative decision making (CDM) process. A suitable method for sharing aircraft trajectories is necessary for this negotiation. Sharing a whole trajectory would require a high amount of bandwidth, and the shared trajectory might become invalid if the weather forecast changed. Instead, a description of the trajectory, decoupled from the weather conditions, could be shared, so that the actual trajectory could be computed from this trajectory description. This trajectory description should be easy to process using a computing program —as some of the CDM processes will be automated— but also easy to understand for a human operator —who will be supervising the process and making decisions. This thesis presents a series of formal languages that can be used for this purpose. These languages provide the means to describe aircraft trajectories during all phases of flight, from push back to arrival at the gate. They can also describe trajectories of both manned and unmanned aircraft, including fixedwing and some rotary-wing aircraft (quadrotors). Some of these languages are tightly interrelated and organized in a language hierarchy. One of the key languages in this hierarchy, the aircraft intent description language (AIDL), had already been developed prior to this thesis. This language was derived from the equations of motion of fixed-wing aircraft, and can provide an unambiguous description of fixed-wing aircraft trajectories. A variant of this language, the quadrotor AIDL (QR-AIDL), is developed in this thesis to allow describing a quadrotor aircraft trajectory with the same level of detail. Then, the intent composite description language (ICDL) is built on top of these two languages, providing more flexibility to describe some parts of the trajectory while leaving others unspecified. The ICDL is used to provide generic descriptions of common aircraft manoeuvres, which can be particularized and combined to form complex descriptions of flight. Another language is built on top of the ICDL, the flight intent description language (FIDL). The FIDL specifies high-level requirements on trajectories —including constraints and objectives—, but can use features of the ICDL to provide arbitrary levels of detail in different parts of the flight. The ICDL and FIDL have been developed in collaboration with Boeing Research & Technology Europe (BR&TE). Also, the mission intent description language (MIDL) has been developed to allow describing missions involving multiple aircraft. This language is based on the FIDL and keeps all its expressive power, while it also provides new semantics for describing mission tasks, mission objectives, and constraints involving several aircraft. In ATM, the movement of aircraft while on the airport surface also has to be monitored and managed. Another formal language has been designed for this purpose, denoted surface movement description language (SMDL). This language does not belong to the language hierarchy described above, and it is based on the clearances used in airport surface operations. Means to express uncertainty and mutability of different parts of the trajectory are also provided. Finally, the applications of these languages to trajectory prediction and mission planning are explored in this thesis. The concept of trajectory language processing engine (TLPE) is used in these two applications. A TLPE is an ATM function whose main input and output are expressed in any of the languages in the hierarchy described in this thesis. A modular trajectory predictor is defined as a combination of multiple TLPEs, each of them performing a small subtask. Special attention is given to the TLPE that builds the horizontal, vertical, and configuration profiles of the trajectory. In particular, a novel method for the generation of the vertical profile is presented. The process of planning a mission can also be seen as a TLPE, where the main input is expressed in the MIDL and the output consists of a number of trajectory descriptions —one for each aircraft available in the mission— expressed in the FIDL. A mixed integer linear programming (MILP) formulation for the problem of assigning mission tasks to the available aircraft is provided. In addition, since finding optimal paths between locations is a key problem to mission planning, a novel path finding algorithm is presented. This algorithm can compute near-shortest paths avoiding all obstacles in an urban environment in very short times. The several formal languages described in this thesis can serve as a standard specification to share trajectory information among different actors in ATM. In combination, these languages can describe trajectories with the necessary level of detail for any application, and can be used to increase automation by exploiting this information using decision support tools (DSTs). Their applications to some basic functions of DSTs, such as trajectory prediction, have been analized.
Resumo:
Considering the well established role of nonclassical HLA-G class I molecules in inhibiting natural killer (NK) cell function, the consequence of abnormal HLA-G expression in malignant cells should be the escape of tumors from immunosurveillance. To examine this hypothesis, we analyzed HLA-G expression and NK sensitivity in human malignant melanoma cells. Our analysis of three melanoma cell lines and ex vivo biopsy demonstrated that (i) IGR and M74 human melanoma cell lines exhibit a high level of HLA-G transcription with differential HLA-G isoform transcription and protein expression patterns, (ii) a higher level of HLA-G transcription ex vivo is detected in a skin melanoma metastasis biopsy compared with a healthy skin fragment from the same individual, and (iii) HLA-G protein isoforms other than membrane-bound HLA-G1 protect IGR from NK lysis. It thus appears of critical importance to consider the specific role of HLA-G expression in tumors in the design of future cancer immunotherapies.
Resumo:
In many organisms nonsense mutations decrease the level of mRNA. In the case of mammalian cells, it is still controversial whether translation is required for this nonsense-mediated RNA decrease (NMD). Although previous analyzes have shown that conditions that impede translation termination at nonsense codons also prevent NMD, the residual level of termination was unknown in these experiments. Moreover, the conditions used to impede termination might also have interfered with NMD in other ways. Because of these uncertainties, we have tested the effects of limiting translation of a nonsense codon in a different way, using two mutations in the immunoglobulin μ heavy chain gene. For this purpose we exploited an exceptional nonsense mutation at codon 3, which efficiently terminates translation but nonetheless maintains a high level of μ mRNA. We have shown 1) that translation of Ter462 in the double mutant occurs at only ∼4% the normal frequency, and 2) that Ter462 in cis with Ter3 can induce NMD. That is, translation of Ter462 at this low (4%) frequency is sufficient to induce NMD.
Resumo:
Ribozyme activity in vivo depends on achieving high-level expression, intracellular stability, target colocalization, and cleavage site access. At present, target site selection is problematic because of unforeseeable secondary and tertiary RNA structures that prevent cleavage. To overcome this design obstacle, we wished to engineer a ribozyme that could access any chosen site. To create this ribozyme, the constitutive transport element (CTE), an RNA motif that has the ability to interact with intracellular RNA helicases, was attached to our ribozymes so that the helicase-bound, hybrid ribozymes would be produced in cells. This modification significantly enhanced ribozyme activity in vivo, permitting cleavage of sites previously found to be inaccessible. To confer cleavage enhancement, the CTE must retain helicase-binding activity. Binding experiments demonstrated the likely involvement of RNA helicase(s). We found that attachment of the RNA motif to our tRNA ribozymes leads to cleavage in vivo at the chosen target site regardless of the local RNA secondary or tertiary structure.
Resumo:
Aquaporins are integral membrane proteins of the tonoplast and the plasma membrane that facilitate the passage of water through these membranes. Because of their potentially important role in regulating water flow in plants, studies documenting aquaporin gene expression in specialized tissues involved in water and solute transport are important. We used in situ hybridization to examine the expression pattern of the tonoplast aquaporin ZmTIP1 in different organs of maize (Zea mays L.). This tonoplast water channel is highly expressed in the root epidermis, the root endodermis, the small parenchyma cells surrounding mature xylem vessels in the root and the stem, phloem companion cells and a ring of cells around the phloem strand in the stem and the leaf sheath, and the basal endosperm transfer cells in developing kernels. We postulate that the high level of expression of ZmTIP1 in these tissues facilitates rapid flow of water through the tonoplast to permit osmotic equilibration between the cytosol and the vacuolar content, and to permit rapid transcellular water flow through living cells when required.
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The relationship between the optimization of the potential function and the foldability of theoretical protein models is studied based on investigations of a 27-mer cubic-lattice protein model and a more realistic lattice model for the protein crambin. In both the simple and the more complicated systems, optimization of the energy parameters achieves significant improvements in the statistical-mechanical characteristics of the systems and leads to foldable protein models in simulation experiments. The foldability of the protein models is characterized by their statistical-mechanical properties--e.g., by the density of states and by Monte Carlo folding simulations of the models. With optimized energy parameters, a high level of consistency exists among different interactions in the native structures of the protein models, as revealed by a correlation function between the optimized energy parameters and the native structure of the model proteins. The results of this work are relevant to the design of a general potential function for folding proteins by theoretical simulations.
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Resting epidermal keratinocytes contain large amounts of interleukin 1 (IL-1), but the function of this cytokine in the skin remains unclear. To further define the role of IL-1 in cutaneous biology, we have generated two lines of transgenic mice (TgIL-1.1 and TgIL-1.2) which overexpress IL-1 alpha in basal keratinocytes. There was high-level tissue-specific expression of transgene mRNA and protein and large quantities of IL-1 alpha were liberated into the circulation from epidermis in both lines. TgIL-1.1 mice, which had the highest level of transgene expression, developed a spontaneous skin disease characterized by hair loss, scaling, and focal inflammatory skin lesions. Histologically, nonlesional skin of these animals was characterized by hyperkeratosis and a dermal mononuclear cell infiltrate of macrophage/monocyte lineage. Inflammatory lesions were marked by a mixed cellular infiltrate, acanthosis, and, in some cases, parakeratosis. These findings confirm the concept of IL-1 as a primary cytokine, release of which is able to initiate and localize an inflammatory reaction. Furthermore, these mice provide the first definitive evidence that inflammatory mediators can be released from the epidermis to enter the systemic circulation and thereby influence, in a paracrine or endocrine fashion, a wide variety of other cell types.
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Esta pesquisa aborda as chamadas políticas de diversidade na educação e sua contribuição para o reconhecimento e a promoção dos direitos humanos e a superação do racismo, do sexismo, da homofobia e das demais desigualdades e discriminações que marcam profundamente a sociedade e a educação brasileiras. Com base nas vozes de gestores/as públicos/as e ativistas da sociedade civil, na análise documental e da execução orçamentária e na experiência política da pesquisadora, é apresentado um balanço sobre os dez anos de existência da Secretaria de Educação Continuada, Alfabetização e Diversidade (Secad), órgão do Ministério da Educação criado no primeiro governo do Presidente Luiz Inácio Lula da Silva. Em especial, buscou-se identificar as provocações e os tensionamentos gerados pelas agendas das diversidades para o atual desenho, funcionamento e institucionalidade das políticas educacionais e sua influência nas concepções de qualidade educacional em disputa nas políticas federais. Essas disputas estiveram presentes nas Conferências Nacionais de Educação e no processo conflitivo de tramitação do novo Plano Nacional de Educação (Lei Federal n. 13.005/2014), analisados neste trabalho. Respaldado por convenções e pelas resoluções internacionais das Conferências da ONU e por normativas nacionais, o debate sobre diferenças ganhou espaço na agenda das políticas educacionais brasileiras. Essa discussão foi impulsionada por movimentos sociais negros, indígenas, LGBTs, feministas, de trabalhadores do campo, de pessoas com deficiências, de quilombolas, ambientalistas e por agendas de fronteira na efetividade do direito humano à educação, como a educação de jovens e adultos, a educação em territórios de alta vulnerabilidade social e a educação de pessoas privadas de liberdade, entre outras. Apresenta-se, neste trabalho, uma contribuição teórica ao debate sobre a relação entre qualidade educacional, diferenças e igualdades, com base nas teorias críticas de justiça social. Discutem-se as possibilidades de a noção da diversidade constituir uma resposta interseccional às múltiplas discriminações e desigualdades que atingem os sujeitos concretos no cotidiano da vida e, especificamente, nas instituições educacionais. Ao final da tese, embasadas na definição do contexto de estratégia política de Stephen Ball e nas contribuições para o aperfeiçoamento das políticas 14 previstas na metodologia de análise das políticas públicas, são apresentadas reflexões comprometidas com a ampliação da capacidade das políticas educacionais no sentido de dar respostas a essas agendas, em uma perspectiva de promoção da justiça na educação no marco dos direitos humanos.
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Hoje em dia com o crescente aumento da exploração de petróleo e gás em águas profundas, há um aumento na demanda por operações offshore envolvendo a cooperação entre unidades flutuantes. Tais operações requerem um alto nível de planejamento e coordenação, o que na maioria dos casos é feito com a troca de informação no nível de operação, com cada unidade flutuante comandada independentemente. Exemplos de operações deste tipo vão desde operações de alívio passando por operações de instalação de equipamento submarino, até operações de pesquisa envolvendo múltiplas unidades flutuantes dotadas de sistema de posicionamento dinâmico (DP). As vantagens do controle cooperativo surgem com a redução do erro da distância relativa durante a manutenção do posicionamento ou durante a execução de manobras de posicionamento conjuntas. No presente trabalho, os conceitos de controle de consenso são aplicados de forma combinada com o sistema DP de cada navio. A influência dos ganhos do controlador cooperativo no sistema como um todo será discutida, utilizando-se técnicas de análise da resposta em frequência. Simulações completas no domínio do tempo e experimentos usando modelos em escala serão utilizados para se demonstrar o funcionamento do controle cooperativo. Todas as simulações serão conduzidas no simulador Dynasim e os ensaios experimentais no tanque de provas da Engenharia Naval da Escola Politécnica da Universidade de São Paulo. Além disso, serão feitas comparações entre os experimentos em tanque de provas e simulações numéricas equivalentes, demonstrando-se a validade dos ensaios numéricos. Será também demonstrado que os requisitos de projetos adotados são atendidos pelos ensaios em tanque de provas. .
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The retina is a very complex neural structure, which performs spatial, temporal, and chromatic processing on visual information and converts it into a compact ‘digital’ format composed of neural impulses. This paper presents a new compiler-based framework able to describe, simulate and validate custom retina models. The framework is compatible with the most usual neural recording and analysis tools, taking advantage of the interoperability with these kinds of applications. Furthermore it is possible to compile the code to generate accelerated versions of the visual processing models compatible with COTS microprocessors, FPGAs or GPUs. The whole system represents an ongoing work to design and develop a functional visual neuroprosthesis. Several case studies are described to assess the effectiveness and usefulness of the framework.
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Wood is a natural and traditional building material, as popular today as ever, and presents advantages. Physically, wood is strong and stiff, but compared with other materials like steel is light and flexible. Wood material can absorb sound very effectively and it is a relatively good heat insulator. But dry wood burns quite easily and produces a great deal of heat energy. The main disadvantage is the high level of combustion when exposed to fire, where the point at which it catches fire is from 200–400°C. After fire exposure, is need to determine if the charred wooden structures are safe for future use. Design methods require the use of computer modelling to predict the fire exposure and the capacity of structures to resist those action. Also, large or small scale experimental tests are necessary to calibrate and verify the numerical models. The thermal model is essential for wood structures exposed to fire, because predicts the charring rate as a function of fire exposure. The charring rate calculation of most structural wood elements allows simple calculations, but is more complicated for situations where the fire exposure is non-standard and in wood elements protected with other materials. In this work, the authors present different case studies using numerical models, that will help professionals analysing woods elements and the type of information needed to decide whether the charred structures are adequate or not to use. Different thermal models representing wooden cellular slabs, used in building construction for ceiling or flooring compartments, will be analysed and submitted to different fire scenarios (with the standard fire curve exposure). The same numerical models, considering insulation material inside the wooden cellular slabs, will be tested to compare and determine the fire time resistance and the charring rate calculation.
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Larry Summers has attracted much attention recently for invoking old theories of secular stagnation to explain the persistence of low interest rates in the recent past. The German economist Carl Christian von Weizsäcker has pointed to a retirement savings glut as the cause for low rates. In the view of Thomas Mayer, however, as expressed in this High-Level Brief, these theses lack both theoretical and empirical support and he offers as an alternative explanation the fall-out from the recent credit boom-bust cycle.