Isolation and identification of molecular partners of the proteins encoded by the Drosophila tumor suppressor gene lethal(2)tumorous imaginal discs

Ziel dieser Arbeit war es, die funktionelle Bedeutung des Drosophila melanogaster tumor suppressor Gens lethal(2)tumorous imaginal discs (l(2)tid) durch die Identifikation von molekularen Partnern der vom Gen kodierten Proteine zu etablieren. Mit dem Screening einer Expressionsbibliothek mittels des Hefe-Di-Hybrid-Systems wurde das Protein Patched (Ptc) als ein neues Tid-bindendes Protein identifiziert. Ptc ist ein Zentralregulator der Hedhehog-Signalkette. Diese ist in der Entwicklung konserviert und in manchen humanen Krebsarten verwickelt. Die Tid/Ptc-Interaktion wurde mittels unabhängigen biochemischen Methoden wie dem GST-pulldown-Test oder der Immunopräzipitation überprüft. Außerdem ergaben funktionelle Studien in tumorosen Imaginalscheiben einen möglichen inhibitorischen Effekt von Tid über die Hh Signaltransduktion.Im letzten Teil dieser Arbeit wurde die Interaktion zwischen Tid und dem E-APC-Protein (Adenomatous polyposis coli) bewiesen. Polakis und seine Gruppe zeigten durch Studien mit dem Hefe-Di-Hybrid-System und in vitro, dass das hTid mit dem APC-Protein interagiert. Um dies auch auf Drosophila-Ebene zu überprüfen, wurden Immunopräzipitation-Studien mit den Drosophila-Gegenstücken durchgeführt. Diese Studien zeigen zum ersten Mal eine direkte Interaktion beider Proteine in vivo.

Der Carrier DcuB als zweiter Sensor des Zweikomponentensystems DcuSR in Escherichia coli

Escherichia coli kann C4-Dicarboxylate und andere Carbonsäuren als Substrate für den aeroben und anaeroben Stoffwechsel nutzen. Die Anwesenheit von C4-Dicarboxylaten im Außenmedium wird über das Zweikomponentensystem DcuSR, bestehend aus der membranständigen Sensorkinase DcuS und dem cytoplasmatischen Responseregulator DcuR, erkannt. Die Bindung von C4-Dicarboxylaten an die periplasmatische Domäne von DcuS führt zu einer Induktion der Zielgene. Hierzu zählen die Gene für den anaeroben Fumarat/Succinat-Antiporter DcuB (dcuB), die anaerobe Fumarase (fumB) und die Fumaratreduktase (frdABCD). Unter aeroben Bedingungen stimuliert DcuSR die Expression des dctA Gens, das für den aeroben C4-Dicarboxylat-Carrier DctA kodiert. Für den Carrier DcuB konnte eine regulatorische Funktion bei der Expression der DcuSR-regulierten Gene gezeigt werden. Die Inaktivierung des dcuB Gens führte bereits ohne Fumarat zu einer maximalen Expression einer dcuB´-´lacZ Reportergenfusion und anderer DcuSR-abhängiger Gene. Diese Stimulierung erfolgte nur in einem dcuS-positiven Hintergrund. DcuB unterscheidet sich damit von den alternativen Carriern DcuA und DcuC, die diesen Effekt nicht zeigten. Mithilfe ungerichteter Mutagenese wurden DcuB-Punktmutanten hergestellt (Thr394Ile und Asp398Asn), die eine Geninduktion verursachten, aber eine intakte Transportfunktion besaßen. Dies zeigt, dass der regulatorische Effekt von DcuB unabhängig von dessen Transportfunktion ist. Durch gerichtete Mutagenese wurde die Funktion einer Punktmutation (Thr394) näher charakterisiert. Es werden zwei Modelle zur Membrantopologie von DcuB und der Lage der Punktmutationen im Protein vorgestellt. Da DcuB seine regulatorische Funktion über eine Interaktion mit DcuS vermitteln könnte, wurden mögliche Wechselwirkungen zwischen DcuB und DcuS als auch DcuR mithilfe von Two-Hybrid-Systemen untersucht. Für biochemische Untersuchungen von DcuB wurde außerdem die Expression des Proteins in vivo und in vitro versucht. Unter aeroben Bedingungen beeinflusst der C4-Dicarboxylat-Carrier DctA die Expression der DcuSR-abhängigen Gene. Eine Mutation des dctA Gens bewirkte eine stärkere Expression einer dctA´-´lacZ Reportergenfusion im Vergleich zum Wildtyp. Diese Expression nahm in einem dcuS-negativen Hintergrund ab, die Succinat-abhängige Induktion blieb jedoch erhalten. Unter anaeroben Bedingungen kann das dctA Gen auch durch Inaktivierung von DcuB induziert werden. Es wird ein Modell vorgestellt, das die Beteiligung beider Carrier an der DcuSR-abhängigen Regulation erklärt.

Diblock copolymer - nanocrystal hybrids for optoelectronic applications

Small, smaller, nano - it is a milestone in the development of new materials and technologies. Nanoscience is now present in our daily lives: in the car industry with self-cleaning surfaces, in medicine with cancer therapies, even our clothes and cosmetics utilize nanoparticles. The number and variety of applications has been growing fast in recent years, and the possibilities seem almost infinite. Nanoparticles made of inorganic materials have found applications in new electronic technologies, and organic nanomaterials have been added to resins to produce very strong but light weight materials.rnThis work deals with the combination of organic and inorganic materials for the fabrication of new, functional hybrid systems. For that purpose, block copolymers were made with a long, solubility-enhancing and semiconducting block, and a short anchor block. They were synthesized by either RAFT polymerization or Siegrist polycondensation. For the second block, an active ester was grafted on and subsequently reacted with the anchor molecules in a polymer analogue reaction. The resulting block copolymers had different properties; poly(para-phenylene vinylene) showed self-assembly in organic solvents, which resulted in gelling of the solution. The fibers from a diluted solution were visible through microscopy. When polymer chains were attached to TiO2 nanorods, the hybrids could be integrated into polymer fibers. A light-induced charge separation was demonstrated through KPFM. The polymer charged positively and the charge could travel along the fibers for several hundred nanometers. Polymers made via RAFT polymerization were based on poly(vinyltriphenylamine). Ruthenium chromophores which carried anchor groups were attached to the second block. These novel block copolymers were then attached to ZnO nanorods. A light-induced charge separation was also demonstrated in this system. The ability to disperse inorganic nanoparticles within the film is another advantage of these block copolymers. This was shown with the example of CdSe tetrapods. Poly(vinyltriphenylamine dimer) with disulfide anchor groups was attached to CdSe tetrapods. These four-armed nanoparticles are supposed to show very high charge transport. A polymer without anchor groups was also mixed with the tetrapods in order to investigate the influence of the anchor groups. It was shown that without them no good films were formed and the tetrapods aggregated heavily in the samples. Additionally, a large difference in the film qualities and the aggregation of the tetrapods was found in the sample of the polymer with anchor groups, dependent on the tetrapod arm length and the polymer loading. These systems are very interesting for hybrid solar cells. This work also illustrates similar systems with quantum dots. The influence of the energy level of the polymer on the hole transport from the polymer to the quantum dots, as well as on the efficiency of QLEDs was studied. For this purpose two different polymers were synthesized with different HOMO levels. It was clearly shown that the polymer with the adjusted lower HOMO level had a better hole injection to the quantum dots, which resulted in more efficient light emitting diodes.rnThese systems all have in common the fact that novel, and specially designed polymers, were attached to inorganic nanocrystals. All of these hybrid materials show fascinating properties, and are helpful in the research of new materials for optoelectronic applications.

Predicting dependencies using domain-based coupling

Software dependencies play a vital role in programme comprehension, change impact analysis and other software maintenance activities. Traditionally, these activities are supported by source code analysis; however, the source code is sometimes inaccessible or difficult to analyse, as in hybrid systems composed of source code in multiple languages using various paradigms (e.g. object-oriented programming and relational databases). Moreover, not all stakeholders have adequate knowledge to perform such analyses. For example, non-technical domain experts and consultants raise most maintenance requests; however, they cannot predict the cost and impact of the requested changes without the support of the developers. We propose a novel approach to predicting software dependencies by exploiting the coupling present in domain-level information. Our approach is independent of the software implementation; hence, it can be used to approximate architectural dependencies without access to the source code or the database. As such, it can be applied to hybrid systems with heterogeneous source code or legacy systems with missing source code. In addition, this approach is based solely on information visible and understandable to domain users; therefore, it can be efficiently used by domain experts without the support of software developers. We evaluate our approach with a case study on a large-scale enterprise system, in which we demonstrate how up to 65 of the source code dependencies and 77% of the database dependencies are predicted solely based on domain information.

Hierarchical self-assembly of the DNA-grafted supramolecular polymers

Conjugation of functional entities with a specific set of optical, mechanical or biological properties to DNA strands allows engineering of sophisticated DNA-containing architectures. Among various hybrid systems, DNA-grafted polymers occupy an important place in modern materials science. In this contribution we present the non-covalent synthesis and properties of DNA-grafted linear supramolecular polymers (SPs), which are assembled in a controllable manner from short chimeric DNA-pyrene oligomers. The synthetic oligomers consist of two parts: a 10 nucleotides long DNA chain and a covalently attached segment of variable number of phosphodiester-linked pyrenes. The temperature-dependent formation of DNA-grafted SPs is described by a nucleation-elongation mechanism. The high tendency of pyrenes to aggregate in water, leads to the rapid formation of SPs. The core of the assemblies consists of stacked pyrenes. They form a 1D platform, to which the DNA chains are attached. Combined spectroscopic and microscopic studies reveal that the major driving forces of the polymerization are π-stacking of pyrenes and hydrophobic interactions, and DNA pairing contributes to a lesser extent. AFM and TEM experiments demonstrate that the 1D SPs appear as elongated ribbons with a length of several hundred nanometers. They exhibit an apparent helical structure with a pitch-to-pitch distance of 50±15 nm. Since DNA pairing is a highly selective process, the ongoing studies are aimed to utilize DNA-grafted SPs for the programmable arrangement of functional entities. For example, the addition of non-modified complementary DNA strands to the DNA-grafted SPs leads to the cooperative formation of higher-order assemblies. Also, our experiments suggest that the fluorescent pyrene core of 1D ribbons serves as an efficient donor platform for energy transfer applications.

Hierarchical self-assembly of the DNA-grafted supramolecular polymers

Conjugation of functional entities with a specific set of optical, mechanical or biological properties to DNA strands allows engineering of sophisticated DNA-containing architectures. Among various hybrid systems, DNA-grafted polymers occupy an important place in modern materials science. In this contribution we present the non-covalent synthesis and properties of DNA-grafted linear supramolecular polymers (SPs), which are assembled in a controllable manner from short chimeric DNA-pyrene oligomers. The synthetic oligomers consist of two parts: a 10 nucleotides long DNA chain and a covalently attached segment of variable number of phosphodiester-linked pyrenes. The temperature-dependent formation of DNA-grafted SPs is described by a nucleation-elongation mechanism. The high tendency of pyrenes to aggregate in water, leads to the rapid formation of SPs. The core of the assemblies consists of stacked pyrenes. They form a 1D platform, to which the DNA chains are attached. Combined spectroscopic and microscopic studies reveal that the major driving forces of the polymerization are π-stacking of pyrenes and hydrophobic interactions, and DNA pairing contributes to a lesser extent. AFM and TEM experiments demonstrate that the 1D SPs appear as elongated ribbons with a length of several hundred nanometers. They exhibit an apparent helical structure with a pitch-to-pitch distance of 50±15 nm. Since DNA pairing is a highly selective process, the ongoing studies are aimed to utilize DNA-grafted SPs for the programmable arrangement of functional entities. For example, the addition of non-modified complementary DNA strands to the DNA-grafted SPs leads to the cooperative formation of higher-order assemblies. Also, our experiments suggest that the fluorescent pyrene core of 1D ribbons serves as an efficient donor platform for energy transfer applications.

Simulation of AC, DC, and AC-DC coupled mini-grids. In search of the most efficient system

Renewable energy hybrid systems and mini-grids for electrification of rural areas are known to be reliable and more cost efficient than grid extension or only-diesel based systems. However, there is still some uncertainty in some areas, for example, which is the most efficient way of coupling hybrid systems: AC, DC or AC-DC? With the use of Matlab/Simulink a mini-grid that connects a school, a small hospital and an ecotourism hostel has been modelled. This same mini grid has been coupled in the different possible ways and the system’s efficiency has been studied. In addition, while keeping the consumption constant, the generation sources and the consumption profile have been modified and the effect on the efficiency under each configuration has also been analysed. Finally different weather profiles have been introduced and, again, the effect on the efficiency of each system has been observed.

Optimizing PV use through active demand side management

With recent technological developments within the field of power conditioning and the progressive decrease of incentives for PV electricity in grid-connected markets, new operation modes for PV systems should be explored beyond the traditional maximization of PV electri city feed-in. An example can be found in the domestic sector, where the use of modern PV hybrid systems combin ed with efficient electrical appliances and demand side management strategies can significantly enhance the PV value for the user. This paper presents an active demand side management system able to displace the consumer’s load curve in response to local (PV hybrid system, user) and external conditions (external grid). In this way, th e consumer becomes an “active consumer” that can also cooperate with others and the grid, increasing even more the PV value for the electrical system.

Las Redes de Petri Interpretadas en el Diagnóstico de Fallos de Sistemas Híbridos. Aplicación a un Helicóptero no Tripulado

La seguridad y fiabilidad de los procesos industriales son la principal preocupación de los ingenieros encargados de las plantas industriales. Por lo tanto, desde un punto de vista económico, el objetivo principal es reducir el costo del mantenimiento, el tiempo de inactividad y las pérdidas causadas por los fallos. Por otra parte, la seguridad de los operadores, que afecta a los aspectos sociales y económicos, es el factor más relevante a considerar en cualquier sistema Debido a esto, el diagnóstico de fallos se ha convertido en un foco importante de interés para los investigadores de todo el mundo e ingenieros en la industria. Los principales trabajos enfocados en detección de fallos se basan en modelos de los procesos. Existen diferentes técnicas para el modelado de procesos industriales tales como máquinas de estado, árboles de decisión y Redes de Petri (RdP). Por lo tanto, esta tesis se centra en el modelado de procesos utilizando redes de petri interpretadas. Redes de Petri es una herramienta usada en el modelado gráfico y matemático con la habilidad para describir información de los sistemas de una manera concurrente, paralela, asincrona, distribuida y no determinística o estocástica. RdP son también una herramienta de comunicación visual gráfica útil como lo son las cartas de flujo o diagramas de bloques. Adicionalmente, las marcas de las RdP simulan la dinámica y concurrencia de los sistemas. Finalmente, ellas tienen la capacidad de definir ecuaciones de estado específicas, ecuaciones algebraicas y otros modelos que representan el comportamiento común de los sistemas. Entre los diferentes tipos de redes de petri (Interpretadas, Coloreadas, etc.), este trabajo de investigación trata con redes de petri interpretadas principalmente debido a características tales como sincronización, lugares temporizados, aparte de su capacidad para procesamiento de datos. Esta investigación comienza con el proceso para diseñar y construir el modelo y diagnosticador para detectar fallos definitivos, posteriormente, la dinámica temporal fue adicionada para detectar fallos intermitentes. Dos procesos industriales, concretamente un HVAC (Calefacción, Ventilación y Aire Acondicionado) y un Proceso de Envasado de Líquidos fueron usados como banco de pruebas para implementar la herramienta de diagnóstico de fallos (FD) creada. Finalmente, su capacidad de diagnóstico fue ampliada en orden a detectar fallos en sistemas híbridos. Finalmente, un pequeño helicóptero no tripulado fue elegido como ejemplo de sistema donde la seguridad es un desafío, y las técnicas de detección de fallos desarrolladas en esta tesis llevan a ser una herramienta valorada, desde que los accidentes de las aeronaves no tripuladas (UAVs) envuelven un alto costo económico y son la principal razón para introducir restricciones de volar sobre áreas pobladas. Así, este trabajo introduce un proceso sistemático para construir un Diagnosticador de Fallos del sistema mencionado basado en RdR Esta novedosa herramienta es capaz de detectar fallos definitivos e intermitentes. El trabajo realizado es discutido desde un punto de vista teórico y práctico. El procedimiento comienza con la división del sistema en subsistemas para seguido integrar en una RdP diagnosticadora global que es capaz de monitorear el sistema completo y mostrar las variables críticas al operador en orden a determinar la salud del UAV, para de esta manera prevenir accidentes. Un Sistema de Adquisición de Datos (DAQ) ha sido también diseñado para recoger datos durante los vuelos y alimentar la RdP diagnosticadora. Vuelos reales realizados bajo condiciones normales y de fallo han sido requeridos para llevar a cabo la configuración del diagnosticador y verificar su comportamiento. Vale la pena señalar que un alto riesgo fue asumido en la generación de fallos durante los vuelos, a pesar de eso esto permitió recoger datos básicos para desarrollar el diagnóstico de fallos, técnicas de aislamiento, protocolos de mantenimiento, modelos de comportamiento, etc. Finalmente, un resumen de la validación de resultados obtenidos durante las pruebas de vuelo es también incluido. Un extensivo uso de esta herramienta mejorará los protocolos de mantenimiento para UAVs (especialmente helicópteros) y permite establecer recomendaciones en regulaciones. El uso del diagnosticador usando redes de petri es considerado un novedoso enfoque. ABSTRACT Safety and reliability of industrial processes are the main concern of the engineers in charge of industrial plants. Thus, from an economic point of view, the main goal is to reduce the maintenance downtime cost and the losses caused by failures. Moreover, the safety of the operators, which affects to social and economic aspects, is the most relevant factor to consider in any system. Due to this, fault diagnosis has become a relevant focus of interest for worldwide researchers and engineers in the industry. The main works focused on failure detection are based on models of the processes. There are different techniques for modelling industrial processes such as state machines, decision trees and Petri Nets (PN). Thus, this Thesis is focused on modelling processes by using Interpreted Petri Nets. Petri Nets is a tool used in the graphic and mathematical modelling with ability to describe information of the systems in a concurrent, parallel, asynchronous, distributed and not deterministic or stochastic manner. PNs are also useful graphical visual communication tools as flow chart or block diagram. Additionally, the marks of the PN simulate the dynamics and concurrence of the systems. Finally, they are able to define specific state equations, algebraic equations and other models that represent the common behaviour of systems. Among the different types of PN (Interpreted, Coloured, etc.), this research work deals with the interpreted Petri Nets mainly due to features such as synchronization capabilities, timed places, apart from their capability for processing data. This Research begins with the process for designing and building the model and diagnoser to detect permanent faults, subsequently, the temporal dynamic was added for detecting intermittent faults. Two industrial processes, namely HVAC (Heating, Ventilation and Air Condition) and Liquids Packaging Process were used as testbed for implementing the Fault Diagnosis (FD) tool created. Finally, its diagnostic capability was enhanced in order to detect faults in hybrid systems. Finally, a small unmanned helicopter was chosen as example of system where safety is a challenge and fault detection techniques developed in this Thesis turn out to be a valuable tool since UAVs accidents involve high economic cost and are the main reason for setting restrictions to fly over populated areas. Thus, this work introduces a systematic process for building a Fault Diagnoser of the mentioned system based on Petri Nets. This novel tool is able to detect both intermittent and permanent faults. The work carried out is discussed from theoretical and practical point of view. The procedure begins with a division of the system into subsystems for further integration into a global PN diagnoser that is able to monitor the whole system and show critical variables to the operator in order to determine the UAV health, preventing accidents in this manner. A Data Acquisition System (DAQ) has been also designed for collecting data during the flights and feed PN Diagnoser. Real flights carried out under nominal and failure conditions have been required to perform the diagnoser setup and verify its performance. It is worth noting that a high risk was assumed in the generation of faults during the flights, nevertheless this allowed collecting basic data so as to develop fault diagnosis, isolations techniques, maintenance protocols, behaviour models, etc. Finally, a summary of the validation results obtained during real flight tests is also included. An extensive use of this tool will improve preventive maintenance protocols for UAVs (especially helicopters) and allow establishing recommendations in regulations. The use of the diagnoser by using Petri Nets is considered as novel approach.

Criterios técnicos del proyecto con módulos tridimensionales ligeros: las casas del Solar Decathlon 2005 y 2007

Este trabajo se ocupa de la construcción modular ligera de viviendas unifamiliares, más concretamente aborda el problema de la definición constructiva de las mismas, habida cuenta de lo poco conocido y publicado de este aspecto en particular. Dado que la construcción es fruto de la evolución, resulta de gran importancia el estudio de los antecedentes para comprender la situación actual, por ello para documentar como corresponde el estado de la técnica y de la investigación se estudiaron los antecedentes desde sus primeras manifestaciones a finales del siglo XIX hasta nuestros días. Contrariamente a las profecías de Le Corbusier y otros, la industrialización no ha llegado a la construcción ni en la forma ni en la medida que era de esperar a principios del siglo XX. Sin embargo y a pesar del relativo “fracaso” de la industrialización en convertirse en la forma mayoritaria de producir edificios, lo cierto es que algunos sistemas altamente industrializados, como lo es la construcción modular ligera, han logrado un lugar en el mercado, que en algunos países resulta cuanto menos significativo. Delimitar el estado de la técnica y la situación actual es crucial en este trabajo, toda vez que la construcción industrializada la realizan empresas, y situarse al margen de las mismas y de su producción, nos guste o no, resulta contrario al sentido común. De este modo se han identificado y documentado ejemplos como el de EE.UU. y Japón, entre otros, que resultan muy ilustrativos tanto desde la evolución como del estado actual. Del estado de la técnica y la investigación también resulta la escasez de publicación de detalles constructivos específicos de la construcción modular ligera. Es obvio que los fabricantes intentan blindarse mediante el secreto industrial, logrando que las soluciones constructivas más interesantes queden fuertemente protegidas dentro del conocimiento reservado de la empresa. De este modo un sistema que alcanza grados de prefabricación del 95%, tan atractivo e interesante para el arquitecto queda reservado al conocimiento de unos pocos dentro de las empresas. La búsqueda de mayor información sobre estos sistemas encuentra un filón indiscutible en los proyectos de las casas del concurso Solar Decathlon. En efecto, debido a las condiciones particulares de este concurso, las casas que compiten deben ser instaladas en un plazo corto, por lo que en su mayoría recurren al método de construcción modular ligera, para lograr correctamente este objetivo. Adicionalmente todos los planos de proyecto incluidos los de detalle, así como memorias de especificaciones técnicas, quedan colgadas en la Web, a libre disposición del público. De este modo se ha considerado interesante, conveniente y útil, aprovechar como fuente documental de este trabajo, aparte de lo obtenido en el estado de la técnica y la investigación, los proyectos de las casas de los concursos 2005 y 2007, que fueron las dos últimas ediciones celebradas cuando se inició esta tesis. De modo similar a esta carencia de detalles publicados se observa igualmente una falta de investigación y de metodologías adecuadas a la misma sobre soluciones y detalles constructivos propios de la construcción modular ligera. Por eso y ante la dificultad de manejar adecuadamente una información disponible pero farragosa, este trabajo ha dedicado una parte importante de su esfuerzo a la creación de una metodología adecuada a este tipo de situación. Hay que destacar que cada proyecto puede constar fácilmente de 60 a casi 200 planos, sin contar las memorias técnicas y otros documentos, tales como galerías de imágenes, etc. Por otra parte resulta útil establecer esta metodología, no solo para esta tesis, sino para futuros trabajos de investigación sobre el tema, toda vez que con posterioridad a 2007 se han celebrado nuevas ediciones del concurso con su correspondiente volumen de información disponible. La metodología del análisis de los proyectos se basa en el diseño y creación de siete fichas tipo que resumen los aspectos fundamentales del proyecto desde el punto de vista constructivo, permitiendo de este modo su rápida visualización y comprensión sin pretender exhaustividad, ya que en caso de querer profundizar en el detalle de la información siempre está el proyecto original para ello. Tras estudiar la información obtenida de los proyectos de las casas de concurso, se contrastan y discuten los resultados para obtener conocimientos de interés para el objetivo propuesto en la tesis. Se comparan los resultados procedentes del estado de la técnica y de la investigación y se obtienen las conclusiones correspondientes. De este modo ha sido posible identificar una serie de criterios técnicos de proyecto de viviendas unifamiliares realizadas mediante construcción modular ligera, que además se organizan en varios niveles, por lo que el resultado es un conjunto de criterios como germen de una futura guía o manual. Como conclusiones fundamentales de la tesis hay que destacar las metodológicas, que habilitan la extensión de este estudio a otros trabajos y la aportación original al conocimiento con la definición de una serie de criterios técnicos de proyecto de viviendas realizadas mediante construcción modular ligera, que además de mejorar esta parte del saber constructivo serán de gran ayuda a los arquitectos de cara a la reducción de errores, que a menudo derivan en mayores costes y plazos, cuando no directamente al abandono del sistema constructivo. El trabajo de la tesis se estructura en siete capítulos a saber: Capítulo 1: Introducción, donde se explica el tema de la tesis, el objetivo principal, las limitaciones y se aportan algunas definiciones. Capítulo 2: Estado de la técnica y la investigación, dentro del cual se presentan los antecedentes desde sus inicios pasando por la primera y segunda mitad del siglo XX y finalizando con el estado actual que recorre los sistemas de EE.UU, Japón, Europa, Escandinavia, centro Europa, Francia, España y algunos ejemplos de Latinoamérica. En el estado de la investigación se presentan las publicaciones tanto en revistas científicas indexadas en el JCR, como publicaciones no indexadas en las que se muestran artículos científicos, comunicaciones a congresos, documentación sobre el concurso Solar Decathlon, libros específicos y libros genéricos sobre construcción modular ligera, tesis doctorales tanto generales como específicas sobre el concurso Solar Decathlon, documentos de idoneidad técnica, páginas Web del concurso Solar Decathlon y finalmente páginas Web de empresas de construcción modular ligera y patentes. También se expresan las conclusiones parciales del capítulo así también como la justificación y los objetivos particulares. Capítulo 3: Metodología, se expone en primer lugar el planteamiento del problema, para luego desarrollar los métodos utilizados para el estudio y clasificación del transporte y de la organización modular, la relación entre la organización modular y distribución espacial, el diseño y contenido de las fichas resumen de los 38 proyectos presentados al SD2005 y SD2007, así también como la información consultada para su realización. Se explica asimismo como se realizaron las tablas comparativas partiendo de la información de las fichas. Finalmente se expresa la manera en que se trató el estudio particular de la junta entre módulos. Capítulo 4: Discusión de resultados, dónde en primer lugar se presenta la clasificación y las estrategias del transporte así también como una clasificación de la organización modular, en segundo lugar se estudian las relaciones que existen entre el sistema modular y la organización espacial. A partir de aquí se muestra el estudio de la estructura de todas las casas del SD2005 y SD2007, según el tipo y material de las zapatas, mostrando tanto tablas cuantitativas como tablas gráficas con fotos. De la misma manera se tratan los forjados, los pilares y las vigas y las cubiertas. Se estudia además el cerramiento opaco y acristalado, la cubierta y el forjado según las capas que lo componen. En el caso de las instalaciones se estudian las que son especiales, descartando las normales en este tipo de casas, como son los paneles fotovoltaicos, placas térmicas o tubos de vacío y los sistemas de almacenamiento de energía, como las baterías, presentando tablas numéricas y gráficas con fotos. El transporte se analiza según corresponda al traslado de la vivienda o al transporte de apoyo, según el tipo y la cantidad utilizado. Con respecto al montaje se diferencia en si se usó grúa o no, y en el caso de no utilizarse se muestran los métodos alternativos. Con respecto a la exploración de la organización modular se presentan la cantidad de módulos que utilizó cada casa, así como también la cuantificación de los sistemas híbridos como son elementos lineales 1D, paneles 2D. Por último se muestra el estudio detallado de la junta entre módulos 3D. Finalmente se realiza la propuesta de un conjunto de criterios técnicos de proyecto organizado en cuatro niveles: 1º nivel de criterios generales, 2º nivel sobre sistemas constructivos, 3º nivel de detalles constructivos y el 4º nivel llamado logística obras previas, transporte y montaje. Capítulo 5: Conclusiones y líneas futuras de investigación, se exponen las conclusiones generales, metodológicas, documentales y por último las de construcción modular. Finalmente se realiza una propuesta de líneas futuras de investigación. Capítulo 6: Bibliografía. Capítulo 7: Anexos, en el que se presentan todas las fichas resumen de las casas realizadas por el autor de esta tesis. Además se incluyen los casos concretos de Voisin en Francia y la TVA (Tennessee Valley Authority) en EE.UU., así como referencias de manuales genéricos de construcción ligera, medidas de transporte en España e información sobre estadísticas de viviendas en la Unión Europea. ABSTRACT This paper deals with Light Modular Construction of houses, and specifically addresses the problem of the constructive definition of the same, given how little known and published this subject has been. Since building construction is the result of evolution, it is of great importance to study its background for understanding the current situation, therefore to document the state of the art and research, its history was studied from its origins dating to the end nineteenth century to our days. Contrary to the prophecies of Le Corbusier and others, industrialization has not reached the construction neither in the form nor to the extent that it was expected in the early twentieth century. However, despite the relative "failure" of industrialization to become the major form of production for buildings, the fact is that some highly industrialized systems, such as the so called Lightweight Modular Construction, have achieved a place in the market, which in some countries is at least significant. To outline the state of the art and the current situation is crucial in this work, since industrialized construction is carried out by companies, and to step away from them and their production, whether we like it or not, it is contrary to common sense. So that, several cases have been identified and documented, such as the US and Japan examples, among others, which are very illustrative both from evolution and the current status. The state of the art and research shows also a shortage of publication of specific construction details of light modular construction. Obviously, manufacturers try to shield themselves by trade secret, making the most interesting constructive solutions remain heavily protected within the reserved knowledge of the company, so a system that reaches levels of 95% prefabrication, so attractive and interesting for the architect It is reserved to the knowledge of a few people inside the companies. The search for more information on these systems finds an invaluable reef in the projects of the Solar Decathlon houses. Indeed, due to the particular conditions of this contest, houses competing must be installed in a short time, so mostly turn to modular construction methods for this purpose to achieve properly this goal. Additionally all levels of project, including detailed and technical specifications reports are published on the Web, freely available to the general public. Thus it was considered interesting, convenient and useful to take advantage as a documentary source of this work, apart from what it were obtained in the state of the art and research, the projects of the houses of the 2005 and 2007 contests, which were the last two editions already held before this thesis was started. Similar to this lack of published details it is also observed a lack of research and methodologies adapted to the solutions and construction details of lightweight modular construction. So that, and facing the difficulty of adequately manage the available but bulky information, this work has devoted a significant part of its effort to create an appropriate methodology for this type of situation. It is noteworthy that each project can easily consist of 60 to almost 200 blueprints, not including technical reports and other documents, such as photo galleries, etc. The methodology of the analysis of the projects is based on the design and creation of seven record sheets that summarize key aspects of the project from a construction point of view, thus allowing quick watching and understanding of the project, without claiming completeness, always keeping for further information the blueprints themselves. After studying the information obtained from the projects of the competition houses, the results are compared and discussed to obtain relevant knowledge according to the objective proposed in the thesis. The results from the state of the art and research are also compared and the conclusions so obtained make possible to identify a number of technical design criteria for single family homes made using lightweight modular construction, which also have been organized at various levels, so the result is a set of criteria as a seed for a future guide or manual. Among the main conclusions of the thesis must be noted the methodological ones, that enable the extension of this study to other works and are an original contribution to knowledge with the definition of a number of technical criteria for housing projects made by light modular construction, what in addition to improving this part of the constructive knowledge will be of great help for the architects to reduce errors that often result in higher costs and delays, if not in abandonment of the construction system itself. The work of the thesis is divided into seven chapters as follows: Chapter 1: Introduction, where the topic of the thesis, the main objective, limitations and the provided definitions is explained. Chapter 2: State of the art and research, within which history is presented from the beginning through the first and second half of the twentieth century and ending with the current systems, that spans from the USA to Japan, Europe, Scandinavia, Central Europe, France, Spain and some Latin American examples. In the state of research publications are presented both on scientific journals indexed in the JCR, and non-indexed publications in which scientific articles, conference papers, information about the Solar Decathlon competition, generic and specific books on light modular construction, articles both general and specific on the Solar Decathlon competition, technical approval documents, dissertations, the Solar Decathlon Web pages and finally lightweight modular construction companies Web pages and patents. The partial conclusions of the chapter as well as the rationale and specific objectives are also expressed. Chapter 3: Methodology first of all exposes the problem statement and then develops the methods used for the study and classification of transportation and modular organization, the relationship between the modular organization and spatial distribution, design and content summary records of the 38 projects submitted to the SD2005 and SD2007, as well as information consulted for its realization. It explains equally how the comparative tables based on information from the chips were made. Finally, the way the particular study of the joint between modules is carried out is also treated. Chapter 4: Discussion, first sorting a classification of transport strategies as well as of modular organization, secondly the relationship between the modular system and the spatial organization studied is presented. From then on, the study of the structure of every house Moreover, to establish this methodology is useful not only for this thesis, but for future research on the subject, since after 2007 there have been held new editions of the contest with a corresponding volume of information available. shown in the SD2005 and SD2007 contests, depending on the type and material of the shoe, showing both quantitative tables as graphic boards with pictures is carried out. Similarly slabs, columns and beams and roofs are treated. Furthermore, the opaque and transparent façades, as well as roof and floor enclosure, according to the layers that compose them are studied. In the case of facilities there have been only addressed that which are of a special type, discarding that considered usual in this type of houses such as photovoltaic panels, thermal panels or vacuum tubes and energy storage systems such as batteries, presenting numerical and graphical tables with photos .The transportation is analyzed depending on it is used to move the house or for additional support, depending on the type and quantity of items used. Regarding assembly it has been made a difference if crane is or not used and if not, what the alternative methods are. With respect to the exploration of the modular organization, the amount of modules used in each house is presented, as well as the quantification of hybrid systems such as linear members as well as 1D, 2D panels. Finally, it has been carried out the detailed study of the joint between 3D modules. Finally it is proposed a set of technical criteria organized into four levels, 1st level of general criteria, 2nd level on building systems, 3rd level construction details and 4th level called previous works and logistics, transportation and assembly. Chapter 5: Conclusions and future research, where general, methodological, documentary and finally modular construction findings are presented. A proposal for further research is done. Chapter 6: Bibliography. Chapter 7: Annexes, in which all the record sheets of houses made by the author of this thesis are presented. Besides the specific cases of Voisin in France and TVA (Tennessee Valley Authority) in the USA, as well as general reference manuals on lightweight construction, transportation dimensions in Spain, and information on housing statistics in the European Union are included.

Cells that register logical relationships among proteins

Two-hybrid methods have augmented the classical genetic techniques biologists use to assign function to genes. Here, we describe construction of a two-bait interaction trap that uses yeast cells to register more complex protein relationships than those detected in existing two-hybrid systems. We show that such cells can identify bridge or connecting proteins and peptide aptamers that discriminate between closely related allelic variants. The protein relationships detected by these cells are analogous to classical genetic relationships, but lend themselves to systematic application to the products of entire genomes and combinatorial libraries. We show that, by performing logical operations on the phenotypic outputs of these complex cells and existing two-hybrid cells, we can make inferences about the topology and order of protein interactions. Finally, we show that cells that register such relationships can perform logical operations on protein inputs. Thus these cells will be useful for analysis of gene and allele function, and may also define a path for construction of biological computational devices.

Functional Domains of Rep2, a Transcriptional Activator Subunit for Res2–Cdc10, Controlling the Cell Cycle “Start”

In the fission yeast Schizosaccharomyces pombe, passage from G1 to S-phase requires the execution of the transcriptional factor complex that consists of the Cdc10 and Res1/2 molecules. This complex activates the MluI cell cycle box cis-element contained in genes essential for S-phase onset and progression. The rep2+ gene, isolated as a multicopy suppressor of a temperature-sensitive cdc10 mutant, has been postulated to encode a putative transcriptional activator subunit for the Res2–Cdc10 complex. To identify the rep2+ function and molecularly define its domain organization, we reconstituted the Res2–Cdc10 complex-dependent transcriptional activation in Saccharomyces cerevisiae. Reconstitution experiments, deletion analyses using one and two hybrid systems, and in vivo Res2 coimmunoprecipitation assays show that the Res2–Cdc10 complex itself can recognize but cannot activate MluI cell cycle box without Rep2, and that consistent with its postulated function, Rep2 contains 45-amino acid Res2 binding and 22-amino acid transcriptional activation domains in the middle and C terminus of the molecule, respectively. The functional essentiality of these domains is also demonstrated by their requirement for rescue of the cold-sensitive rep2 deletion mutant of fission yeast.

Sistemas hídricos do Jardim Botânico do Estado de São Paulo: uma experiência em educação para o meio ambiente

O presente trabalho foi desenvolvido para compor um modelo de educação ambiental no Jardim Botânico do Estado de São Paulo, localizado no Parque Estadual Fontes do Ipiranga (PEFI). Devido ao seu propósito educacional, o projeto educativo foi desenvolvido com o apoio de dois professores do ensino básico e de pesquisadores do Núcleo de Educação Ambiental do Jardim Botânico do Estado de São Paulo e das Seções de Ecologia e Ficologia do Instituto de Botânica. O modelo considerou quatro sistemas hídricos do PEFI, a Nascente do Riacho do Ipiranga e os Lagos do Jardim Botânico \"Nascentes, Ninféias e Bugios\", como base para a construção de instrumentos ordenadores de visitas monitoradas de professores do ensino básico e seus alunos ao Jardim Botânico. Como base do projeto, abordou-se o tema meio ambiente como transversal às disciplinas clássicas do sistema educacional brasileiro e empreenderam-se as ações do projeto através do conhecimento de diferentes profissionais sobre o meio ambiente que compuseram o grupo de trabalho e, ao mesmo, buscou-se integrar as ações às reais necessidades dos docentes do Ensino Básico. Nesse sentido, foi essencial a participação dos dois professores de escolas do entorno do Jardim Botânico nas etapas organizacionais do projeto. A caracterização da área de estudo baseou-se em seus aspectos históricos e físicos, bem como na qualidade sanitária dos corpos aquáticos considerados. As determinações físicas-químicas e microbiológicas revelaram uma condição sanitária satisfatória das águas, contudo os valores de coliformes fecais e totais encontrados nos lagos sugerem contaminação de origem animal, comum à região. A produtividade primária também foi determinada para cada corpo aquático, encontrando-se baixos valores de clorofila a nos três lagos e na nascente. Os tipos de algas microscópicas observadas foram os gêneros Chrysochromulina, Ankistrodesmus, Scenedesmus, Cosmarium, Navicula, Eunotia, Cryptomonas, Closterium, Gomphonema, Pinnularia, Pleurotaenium, Arthrospira, Oedogonium, Euastrum, Monoraphidium, Staurastrum, Pleurotaenium, Mallomonas e Chlamydomonas. De acordo com a tabela Carlson (Índice de Estado Trófico) a Nascente e o Lago das Nascentes são considerados oligotróficos, enquanto que o Lago dos Bugios e o das Ninféias, mesotróficos. A primeira ação do projeto, um curso teórico-prático, buscou-se integrar os professores do ensino básico em temas relacionados ao meio ambiente, particularmente naqueles de maior relevância aos profissionais atuantes no Jardim Botânico. Posteriormente, os professores participaram de ações para formatação de instrumentos educativos para visitação monitorada ao jardim, que resultaram na elaboração de um manual de visitação do professor, contendo um mapa da área de visitação e explicações de placas sinalizadoras. Para o núcleo de educação ambiental do Jardim Botânico foi disponibilizada a estrutura básica de um curso de capacitação de professores do ensino básico em temas sobre o meio ambiente, bem como a documentação fotográfica da área estudada, os critérios para a admissão de visita monitorada e os procedimentos para adequação do local da Nascente do Riacho do Ipiranga à visitação pública. Finalmente, deve-se ressaltar que o tema escolhido para a realização dessa dissertação de mestrado foi originado da percepção dos pesquisadores do Instituto de Botânica em destacar a importância do Jardim Botânico através de seus sistemas hídricos, e como poderiam ser utilizados como instrumento educativo nas escolas de São Paulo. Nesse sentido, optou-se por enriquecer o clássico foco dado aos jardins botânicos em todo o mundo, no qual a maioria dos visitantes apenas observa a vegetação local e exótica. Assim, pode-se afirmar que o trabalho empreendido se constituirá em uma contribuição ao professor com a missão de inserir a dimensão ambiental nas mais variadas disciplinas do ensino básico.

Controlador integrado baseado em conceito híbrido e controle geométrico para embarcações posicionadas dinamicamente.

O conceito de controle híbrido é aplicado à operação de alívio entre um FPWSO e um navio aliviador. Ambos os navios mantêm suas posições e aproamentos pelo resultado da ação do seu Sistema de Posicionamento Dinâmico (SPD). O alívio dura cerca de 24 horas para ser concluído. Durante este período, o estado de mar pode se alterar e os calados estão sendo constantemente alterados. Um controlador híbrido é projetado para permitir modificacões dos parâmetros de controle/observação se alguma alteração significante do estado de mar e/ou calado das embarcações ocorrer. O principal objetivo dos controladores é manter o posicionamento relativo entre os navios com o intuito de evitar perigosa proximidade ou excesso de tensão no cabo. Com isto em mente, uma nova estratégia de controle que atue integradamente em ambos os navios é proposta baseda em geometria diferencial. Observadores não lineares baseados em passividade são aplicados para estimar a posição, a velocidade e as forças externas de mares calmos até extremos. O critério para troca do controle/observação é baseado na variação do calado e no estado de mar. O calado é assumido conhecido e o estado de mar é estimado pela frequência de pico do espectro do movimento de primeira ordem dos navios. Um modelo de perturbação é proposto para encontrar o número de controladores do sistema híbrido. A equivalência entre o controle geométrico e o controlador baseado em Multiplicadores de Lagrange é demonstrada. Assumindo algumas hipóteses, a equivalência entre os controladores geométrico e o PD é também apresentada. O desempenho da nova estratégia é avaliada por meio de simulações numéricas e comparada a um controlador PD. Os resultados apresentam muito bom desempenho em função do objetivo proposto. A comparação entre a abordagem geométrica e o controlador PD aponta um desempenho muito parecido entre eles.

Optimização de um sistema híbrido off-grid PV, Gerador Diesel e Baterias

Tese de mestrado integrado, Engenharia da Energia e do Ambiente, Universidade de Lisboa, Faculdade de Ciências, 2016