Desarrollo sostenible de la construcción naval: dinámica evolutiva y eco-innovación en el sector de reparaciones europeo

El impacto ambiental directo de la construcción naval, que se refiere a la construcción, mantenimiento y reparación de buques, no es de ninguna manera pequeño. La construcción de buques depende de un gran número de procesos que por sí mismos constituyen un riesgo significativo de daño medio ambiental en el entorno de los astilleros y que conducen a emisiones significativas de gases de efecto invernadero. Además, la construcción naval utiliza algunos materiales que no sólo puede llevar a graves consecuencias para el daño ambiental durante su producción y su uso en el proceso de construcción de la nave, sino también posteriormente durante la reparación de buques, el funcionamiento y las actividades de reciclaje. (OECD 2010) El impacto ambiental directo de la construcción naval constituye de por sí, un desafío importante para la industria. Pero este impacto no queda limitado a su entorno inmediato, aunque la Construcción Naval no es directamente responsable de la repercusión en el medio ambiente de la operación y el reciclaje de buques comerciales, si es una parte integral de estas actividades. (OECD 2010) En esta tesis se sugiere que el sector de la construcción naval puede y debe aceptar sus responsabilidades ambientales no solo en el entorno del astillero; también en la operación de los buques, sus productos; tomando conciencia, a través de un enfoque de ciclo de vida, del desempeño ambiental de la industria en su conjunto. Es necesario intensificar esfuerzos a medida que el impacto ambiental de la industria es cada vez más visible en el dominio público, en pro de un crecimiento verde que permita aumentar la capacidad de actividad o producción económica al tiempo que reduce o elimina, los impactos ambientales. Este será un imperativo para cualquier futura actividad industrial y exigirá naturalmente conocimiento ambiental intrincado perteneciente a todos los procesos asociados. Esta tesis - aprovechando como valiosa fuente de información los desarrollos y resultados del proyecto europeo: “Eco_REFITEC”. FP7-CP-266268, coordinado por el autor de esta Tesis, en nombre de la Fundación Centro Tecnológico SOERMAR - tiene como primer objetivo Investigar la interpretación del concepto de Construcción Naval y Transporte Marítimo sostenible así como las oportunidades y dificultades de aplicación en el sector de la Construcción y reparación Naval. Ello para crear o aumentar el entendimiento de la interpretación del concepto de transporte marítimo sostenible y la experiencia de su aplicación en particular en los astilleros de nuevas construcciones y reparación. Pretende también contribuir a una mejor comprensión de la industria Marítima y su impacto en relación con el cambio climático, y ayudar en la identificación de áreas para la mejora del desempeño ambiental más allá de las operaciones propias de los astilleros; arrojando luz sobre cómo puede contribuir la construcción naval en la mejora de la eficiencia y en la reducción de emisiones de CO2 en el transporte marítimo. Se espera con este enfoque ayudar a que la Industria de Construcción Naval vaya abandonando su perspectiva tradicional de solo mirar a sus propias actividades para adoptar una visión más amplia tomando conciencia en cuanto a cómo sus decisiones pueden afectar posteriormente las actividades aguas abajo, y sus impactos en el medio ambiente, el cambio climático y el crecimiento verde. Si bien cada capítulo de la tesis posee su temática propia y una sistemática específica, a su vez retoma desde una nueva perspectiva cuestiones importantes abordadas en otros capítulos. Esto ocurre especialmente con algunos ejes que atraviesan toda la tesis. Por ejemplo: la íntima relación entre el transporte marítimo y el sector de construcción naval, la responsabilidad de la política internacional y local, la invitación a buscar nuevos modos de construir el futuro del sector a través de la consideración de los impactos ambientales, económicos y sociales a lo largo de ciclo de vida completo de productos y servicios. La necesidad de una responsabilidad social corporativa. Estos temas no se cierran ni terminan, sino que son constantemente replanteados tratando de enriquecerlos. ABSTRACT The direct environmental impact of shipbuilding, which refers to construction, maintenance and repair of vessels, is by no means small. Shipbuilding depends on a large number of processes which by themselves constitute significant risks of damage to the shipyards‘ surrounding environment and which lead to significant emissions of greenhouse gases. In addition, shipbuilding uses some materials which not only may carry serious implications for environmental harm during their production and usage in the ship construction process, but also subsequently during ship repairing, operation, and recycling activities. (OECD 2010) The direct environmental impact of shipbuilding constitutes a major challenge for the industry. But this impact is not limited on their immediate surroundings, but while not being directly responsible for the impact on the environment from the operation and final recycling of commercial ships, shipbuilding is an integral part of these activities. (OECD 2010) With this in mind, this thesis is suggested that the shipbuilding industry can and must take up their environmental responsibilities not only on their immediate surroundings, also on the ships operation, becoming aware through a “life cycle” approach to ships, the environmental performance of the industry as a whole. As the environmental impact of the industry is becoming increasingly visible in the public domain much more effort is required for the sake of “green growth” which implies the ability to increase economic activity or output while lowering, or eliminating, environmental impacts. This will be an imperative for any future industrial activity and will naturally demand intricate environmental knowledge pertaining to all associated processes. This thesis making use as a valuable source of information of the developments and results of an European FP7-collaborative project called "Eco_REFITEC, coordinated by the author of this thesis on behalf of the Foundation Center Technology SOERMAR, has as its primary objective to investigate the interpretation of a sustainable Shipbuilding and Maritime Transport concept and the challenges and opportunities involved in applying for the Shipbuilding and ship repair Sector. It is done to improve the current understanding regarding sustainable shipping and to show the application experience in shipbuilding and ship repair shipyards. Assuming that sustainability is more than just an act but a process, this academic work it also aims to contribute to a much better understanding of the maritime industry and its impact with respect to climate change, and help in identifying areas for better environmental performance beyond the shipyard's own operations; shedding light on how shipbuilding can contribute in improving efficiency and reducing CO2 emissions in shipping. It is my hope that this thesis can help the Shipbuilding Industry to abandon its traditional perspective where each simply looks at its own activities to take a broader view becoming aware as to how their decisions may further affect downstream activities and their impacts on the environment, climate change and green growth. Although each chapter will have its own subject and specific approach, it will also take up and re-examine important questions previously dealt with. This is particularly the case with a number of themes which will reappear as the thesis unfolds. As example I will point to the intimate relationship between the shipping and shipbuilding industry, the responsibility of international and local policy, the call to seek other ways of building the future of the sector through the consideration of the environmental, economic and social impacts over the full life cycle of the products and services, the need for a corporate social responsibility. These questions will not be dealt with once and for all, but reframed and enriched again and again.

Lo público en lo privado : la calle elevada como catalizador del encuentro colectivo

En la actualidad, el individualismo social originado por las nuevas sociedades de la información, la crisis económica, y, aún más acusado por aquella arquitectura que busca la máxima rentabilidad, ha provocado una creciente incomunicación entre las personas que comparten los mismos espacios residenciales. La presente tesis doctoral aborda el estudio del espacio común dentro del proyecto de vivienda colectiva, desde la noción de espacio intermedio, entendido como frontera espacial –entre el espacio público de la ciudad y la atmosfera íntima de la vivienda– donde tienen lugar los encuentros sociales entre residentes. El principal objetivo de la investigación es rescatar y poner en valor principios básicos del diseño arquitectónico –muchos de ellos en desuso– que mejoran la calidad del espacio colectivo y favorecen la aparición de una atmósfera agradable para el encuentro interpersonal, así como un intento de codificación de estos aspectos, atendiendo a criterios conceptuales que definen el espacio público contemporáneo. La hipótesis de partida se basa en la consideración del proyecto de viviendas como modelo análogo al proyecto urbano de la ciudad –con el que guarda sugerentes similitudes–, y los espacios intermedios como espacio de referencia que pone en valor aquello que liga – integrado en el propio proyecto–. Este supuesto básico ha permitido a diferentes autores poner en relación fecunda el diseño de la casa, el edificio y la ciudad, con el individuo, la comunidad y la sociedad; y ha posibilitado plantear la trasposición de conceptos arquitectónicos fructíferos, en relación con el espacio público –usando como representante la idea de calle elevada–, que ahora pueden enriquecer el entorno privado del edificio. Sirviéndose de instrumentos específicos de la disciplina arquitectónica, la tesis trata de explicar la devaluación a nivel social de los espacios colectivos, derivada de la construcción en altura y del uso excesivo de la estandarización en el proyecto de viviendas a partir del período de postguerra, e intensificada en el postmodernismo por la revolución de las comunicaciones. Se puede así reorientar el diseño de los proyectos de vivienda colectiva no como una simple agrupación de viviendas que favorecen la incomunicación, sino tratando de impulsar unas edificaciones que potencien la comunicación interpersonal. La reunión y reagrupación de un vocabulario arquitectónico específico, acorde con la complejidad social intrínseca en los espacios colectivos, amplia el alcance de los conceptos tratados. Dicha terminología se basa principalmente en mecanismos de cualificación espacial y permite evaluar la dimensión colectiva de la esfera semipública en el edificio de viviendas. ABSTRACT Currently, the social individualism caused by the new societies of information, the economic crisis and even more by architecture, which looks for maximum profitability, has led to a rising lack of communication between people that share the same living space. This doctoral thesis deals with the study of common space, which is based on the notion of in-between space, within the collective housing project. It is considered as a spatial frontier –between the public space of the city and the intimate atmosphere of the housing– where social contact between residents takes place. The main purpose of this research is to collect basic principles of architectural design –many of them no longer in use– that improve the quality of collective space. Following conceptual criteria that define the contemporary public space, they encourage the emergence of a pleasant atmosphere for interpersonal meeting as well as an attempt to codify these aspects. The initial hypothesis is based on the consideration of the housing project as a model analogous to the urban project of the city, with which it bears some similarities. Furthermore, it is based on the in-between spaces as space of reference that valuates what binds – integrated in the project-. That basic supposition has served various authors to connect the design of the house, the building and the city with the individual, the community and the society. In addition –regarding public space and using the idea of the street in the sky as representative–, it has enabled to bring up the transposition of productive architectural concepts, which can now enrich the private environment of the building. Using specific architectural instruments, this thesis deals with explaining the social devaluation of collective spaces. It has resulted from the high rise building construction as well as from the excessive standardization in the housing project since the postwar period and is intensified by the communications revolution in postmodernism. Thus, it is possible to change the design of collective housing projects not as a mere cluster of dwellings that enhance isolation, but by trying to promote buildings that improve interpersonal communication. The collection and regrouping of a specific architectural vocabulary, in accordance with the social complexity inherent in collective spaces, expands the scope of the discussed concepts. Such terminology is mainly based on qualifying spatial strategies and allows to assess the collective dimension of the semi-public sphere in dwelling.

Habitar al borde del agua : villas del siglo XX

Las casas del siglo XX construidas al borde del mar –escenario único y origen de su expresión- siguen la topografía del suelo que habitan en su descenso al agua, y organizan espacios que obtienen miradas al horizonte marino. El horizonte nos confronta a lo desconocido. La vista del mar incita al deseo de recorrerlo, al deseo de viajar. Con origen en el otium romano, la presencia del agua define un modo de vida apacible –epicúreo- que los viajeros de la arquitectura -que nos acompañan en la tesis- traducen en sus refugios más íntimos. Experimentan con los cambios en los conceptos y en las técnicas, que se trasladan fácilmente a la arquitectura de la casa al borde del agua desde los inicios del Movimiento Moderno. Sus espacios del habitar frente al mar nos permiten descubrir estrategias comunes en sus respuestas más modernas. El arquitecto ante el proyecto frente al mar llega a la ubicación elegida, mira hacia el horizonte, y desplazándose ladera arriba se coloca en un punto estratégico que elige; después, delante de su tablero, recorre el camino inverso, coloca el lugar y sobre él dibuja los elementos que configurarán los espacios de la casa buscando esa mirada al mar. Las situaciones y ubicaciones posibles son infinitas; se definen algunas consonancias espaciales comunes en los suelos que se ocupan debidas a la presencia del mar que asociamos entre sí. De la comparación entre todas las casas -emblemas del XX-, surgen múltiples variantes de la mirada y de espacios al abierto, y modos de fabricar entornos con criterios comunes para dominar la visión del mar. Interiores que se abren al panorama, espacios cuyas ventanas buscan su mirada en la extensión del horizonte, rescisiones y aperturas. Reconocemos condicionantes en el territorio a los que las villas responden, categorías arquitectónicas que dan respuesta frente al mar en la búsqueda del moderno, la topografía, la mirada y el espacio al abierto. Las casas comparten la idea del dominio del paisaje desde el punto más alto, y en algunos suelos se invierte la tipología por la topografía, confirmando así un criterio común basado en la lectura del suelo como consecuencia de la búsqueda del espacio de la mirada. Los espacios al abierto se significan en todas ellas, son espacios al -aire libre- abiertos, unos envueltos, otros porticados, puertas del horizonte que se abren al exterior, en el techo de la casa, otros cubiertos y abiertos, espacios entre interior y exterior, en plataformas con bancales o patios envolventes, recintos o habitaciones abiertas. Descubrimos un logro del XX en los espacios positivos o negativos que traducen o juegan con el entorno, que ocupan o sustraen de los contornos construidos y que obtienen espacios intermedios en la búsqueda de la relación con el mar. Las herramientas que se utilizan son los dibujos de los autores, de las casas visitadas, el elenco de viajeros y sus viajes, el conocimiento desde el estudio de los proyectos. A través de la comparación por aproximaciones parciales, los dibujos nos definen la mirada al mar, el modo de ocupación y la forma de relación con el paisaje. La arquitectura del habitar frente al mar en el XX, hecha para y por arquitectos, topografía el suelo y construye la mirada, fabricando espacios al abierto en la relación entre la casa y el entorno marítimo. ABSTRACT Houses of the 20th century built by the sea – a unique setting which gives rise to their expression – follow the topography of the land they occupy in its descent towards the sea, and they organize spaces which give views of the maritime horizon. The horizon brings us face to face with the unknown. The sea view provokes a desire to cross it, to travel. The presence of the sea defines a peaceful, epicurean way of life, with origins in the Roman otium, which architectural travellers – who accompany us through the thesis – translate into their most intimate retreats. They experiment with changes in concepts and techniques, which are easily transferred to the architecture of the seaside house since the beginnings of the Modern Movement. Their living spaces allow us to discover common strategies in the most modern responses. The architect with a seaside project arrives at the site, looks towards the horizon, then walks uphill and chooses a strategic point; then with his drawing board he retraces his steps, he sets the position and then draws in the elements that make up the house that seeks a sea view. The number of potential situations and locations is infinite; certain common spatial accordances are defined in land which is occupied due to the presence of the sea. Comparison of all the houses – 20th century emblems – throws up multiple variations of view and open spaces, and ways of creating settings with common criteria so as to command the vision of the sea. Interiors which open up to the panorama, spaces whose windows seek their view in the expanse of the horizon, openings and closures. We recognise determinant factors in the territory to which the villas respond, architectural categories which give a seaside solution to the search for the modern, the topography, the view, and the open space. The houses share the idea of dominating the landscape from the highest point, and in some areas typology and topography are inverted, thus confirming a common criteria based on the reading of the ground as a conse quence of the search for the view space. Open spaces stand out in all the villas – spaces open to the outdoor air - some are wrapped, some arcaded, doors to the horizon which open up to the exterior, on the roof of the house. There are open and covered spaces, spaces between the exterior and interior, on platforms with banks and surrounding patios, enclosures and open rooms. We discover an achievement of the 20th century in the positive and negative spaces which translate and play with the setting, which occupy or are extracted from built contours and which obtain intermediate spaces in the search for the relationship with the sea. The tools used are the author’s drawings of the houses visited, the cast of travelling companions and their travels, the knowledge gained from study of the projects. Through comparison by means of partial approaches, the drawings define the view of the sea, the occupation mode and the way of relating to the landscape. Architecture for living by the sea in the 20th century, carried out both by and for the architects, shapes the land and constructs the view, creating open spaces in the relationship between the house and the sea surroundings.

Lo intermedio como lugar : lo intersticial, lo fronterizo y lo impreciso en la arquitectura contemporánea

1. Nuestra investigación se centra en el estudio de los “ámbitos o espacios intermedios” en momentos de la arquitectura contemporánea, en un período de transición entre finales de los años 80 y nuestros días. Pretendemos analizar cómo se presenta el ámbito intermedio en el objeto o lugar arquitectónico y su función o relación con el entorno cercano, desde el proyecto hasta su repercusión en la experiencia vivida. 2. El concepto de intermedio entendido como ámbito o espacio intersticial, fronterizo y ambivalente (o multivalente), atraviesa en la presente investigación el campo de la etimología, de la ciencia, del pensamiento y del arte, para detenerse en la arquitectura actual, llevándose a cabo una indagación concreta en el proceso de concepción, en la propuesta formal, espacial y funcional, así como en la percepción y demás experiencias o vivencias en el lugar arquitectónico. 3. En primer lugar nos proponemos estudiar y explorar el concepto de “intermedio” en sus características, funciones y ubicaciones dentro de la cultura contemporánea, detectando sus antecedentes culturales más importantes, para a continuación aplicarlo reflexivamente a obras significativas de arquitectura. La investigación se enfoca en la modalidad figural (imprecisa) de lo intermedio (en alusión al “figural” deleuziano), cuyas metáforas base son lo translúcido, la penumbra y la espuma, vinculadas al actual paradigma cultural de la complejidad (C. Jencks). A continuación se analizan las relaciones o implicaciones de dicha modalidad figural en obras de arquitectos como Eisenman, Holl, Tschumi, Ito, Fujimoto, Van Berkel y Bos, y Siza, comprobando su valor operativo y poniendo de manifiesto su sentido e importancia en la actualidad y en la disciplina. El estudio tiene como trasfondo el concepto de in-between de la obra de Eisenman, y utiliza conceptos de la filosofía del límite de Trías como algunos de sus principales fundamentos. 4. En cada una de las obras de arquitectura presentadas esta modalidad imprecisa se traduce en un ámbito o espacio intermedio figural específico. Son obras en las que real y virtual, matricial y objetual, arquitectura y territorio, existente y nuevo, colectivo e individual, social e íntimo, interior y exterior, y demás usos y funciones se entrelazan o funden. Son arquitecturas cuya geometría ya no se encuentra tan determinada por aspectos bi o tridimensionales del dibujo o de la construcción. A través de la manipulación de estas geometrías más o menos complejas, dinámicas e intersticiales, estas obras reflejan (o intuyen) las premisas del actual Zeitgeist: un cambio de esquemas de objetos a relaciones basado en un pensamiento más holístico, transdisciplinar, sistémico o complejo (E. Morin), y una nueva conciencia colectiva sobre la realidad que anuncia la crisis de la percepción, el cambio de paradigma y nuevos valores (F. Capra). Emerge otra forma de sentir y percibir el mundo, los lugares y los espacios, que poco a poco está cambiando el modo de pensar y dibujar la arquitectura, y consecuentemente de interaccionar con ella. 5. El sentido del espacio intermedio figural, emerge en las cada vez más complejas cualidades morfológico-espaciales y funcionales de una parte importante de la arquitectura actual. Arquitectura, geometría y tecnología informática están más entrelazadas que nunca, y de un modo más libre, para así poder indagar en nuevas formas de pensar y crear lugares en que los “espacios intermedios” que investigamos son un testimonio. El espacio arquitectónico se vuelve afectivo e interactivo, un lugar intermedio figural, formalizado por espacialidades intersticiales que parecen aludir a lo líquido, donde la forma es más matriz que configuración, el espacio más espacialidad que compartimiento, la función más versátil y multivalente que específica o autónoma, y la perplejidad, la imaginación y la evocación se entrecruzan y vuelven patentes. 6. La arquitectura de los lugares intermedios valora no solamente el “plano”, la “configuración” y la “transparencia”, sino sobre todo el “espacio” y la “relación” en sus interrelaciones con los usuarios y demás contextos. De la identidad a la relacionalidad, de la representación a la presentación, es una arquitectura que propone “otros espacios” que más que espacios son topografías y espacialidades intersticiales, de tensión, transición, transformación, relación, intercambio e interacción. Son lugares intermedios reales y virtuales, que se sirven tanto de la morfología como de la topología para conquistar nuevas espacialidades, pretendiendo salir de la estricta operación de “forming”/”morphing”/formación/”conformación” para explorar la de “spacing”/espaciamiento. Son lugares que se basan en conceptos como la zona de indiscernibilidad de Deleuze, la imagen- flujo de Buci-Glucksmann, la suspensión-entre de Sloterdijk, o el espacio no-objetivo al que se refiere José Gil, plasmándose en la arquitectura contemporánea como tropos de lo intersticial. 7. Son los lugares intermedios de la arquitectura actual, de espacialidad háptica, más “afectiva”, generativa e interactiva, donde ese ámbito intermedio transforma y es transformado, “afecta”, “con-mueve” (nos hace mover/accionar) y evoca otros lugares, otras posibilidades de espacio habitable u ocupable. Emerge lo intermedio como lugar, algo que anuncia y exhibe, de manera paradigmática y manifiesta, el monumento de Eisenman en Berlín. ABSTRACT 1. Our research focuses on the study of “in-between spaces or environments” at particular times in contemporary architecture, in a transition period from the end of the 1980’s through to today. We aim to analyse how in-between environments are presented in the architectural object or place and their function or their relationship with the nearby surroundings, from the project through to the experience had. 2. In this research, the in-between concept, understood as an interstitial, border and ambivalent (or multi-purpose) environment or space, is assessed from the viewpoint of etymology, science, thought and art, to conclude in current architecture, with specific exploration of the conception process, of the formal, spatial and functional proposal, together with the perception and other experiences in the architectural place. 3. Firstly, we aim to study and explore the “in-between” concept as regards its features, functions and locations within contemporary culture, revealing its most important cultural background, to then apply it reflexively to important works of architecture. The research focuses on the figural (imprecise) mode of in-between (in allusion to Deleuze’s “figural” approach) whose base metaphors are the translucent, semi-darkness and foam, linked to the current cultural paradigm of complexity (C. Jenks). We then go on to assess the relationships or implications of said figural mode in architectural works by Eisenman, Holl, Tschumi, Ito, Fujimoto, Van Berkel and Bos, and Siza, verifying their operational worth and revealing their meaning and importance today and in this discipline. The backdrop of the study is the in-between concept of Eisenman’s work and it also uses concepts from the philosophy of the limit of Trías as its main underpinnings. 4. In each of the architectural works presented, this imprecise mode is translated into a specific in-between environment or space. They are works in which real and virtual, matricial and objectual, architecture and territory, existing and new, collective and individual, social and intimate, interior and exterior and other uses and functions all intertwine or blend together. They are architectures the geometry of which is not so much determined by bi- or tri-dimensional aspects of the drawing or construction. Through the manipulation of these more or less complex, dynamic and interstitial geometries, these works reflect (or insinuate) the premises of the current Zeitgeist: A change in the scheme of objects to relationships towards a more holistic, transdisciplinary, systemic or complex thought (E. Morin), and a new collective conscience about the reality which announces the crisis of perception, the change in paradigm and new values (F. Capra). Another way of feeling and perceiving the world, places and spaces, which little by little is changing the way of thinking and drawing architecture and hence of interacting with it. 5. The meaning of figural in-between space emerges in the increasingly complex, morphological-spatial and functional qualities of a large part of architecture today. Architecture, geometry and computer technology are more than ever intertwined in a freer way to inquire into other ways of thinking and making places, where the “in-between” spaces we research are a testimony. Architectural space becomes affective and interactive, a figural in-between place, formed by interstitial spatiality which seems to allude to something liquid, where shape is more matrix than configuration, space more spatiality than compartment, its function more versatile and multi-purpose than specific or autonomous, and perplexity, imagination and evocation criss-cross each other, becoming obvious. 6. The architecture of in-between places values not only “flat”, “configuration” and “transparency”, but above all “space” and the “relationship” in its interrelations with users and other contexts. From identity to relationality, from representation to presentation, it is an architecture that proposes “other spaces”, which more than spaces are topographies and interstitial spatialities, of tension, transition, transformation, relation, exchange and interaction. They are real and virtual in-between places, that take in both morphology and topology to conquer new spatialities, aiming to depart from the strict “forming”/”morphing” operation in order to explore “spacing”. They are places that derive both from Deleuze’s indiscernibility zone concept, from Buci-Glucksmann´s image-flow concept, from Sloterdijk’s suspension-between, and from the non-objective space referred to by José Gil, and they are embodied in contemporary architecture as tropes of the interstitial. 7. They are the in-between places in architecture today, of haptic spatiality, more “affective”, generative and interactive, where that in-between environment transforms and is transformed, “affects”, “moves” (it makes us move/act) and evokes other places, other possibilities of habitable or occupiable space. The in-between emerges as a place, which paradigmatically and declaredly, Eisenman´s monument in Berlin announces and exhibits. RESUMO 1. A investigação centra-se no estudo dos “âmbitos ou espaços intermédios” em momentos da arquitectura contemporânea, no período de transição entre os finais dos anos 80 e os nossos dias. Pretende-se analisar como se apresenta o âmbito intermédio no objecto ou lugar arquitectónico e sua função ou relação com o meio envolvente, desde o projecto até à sua repercussão na experiencia vivenciada. 2. O conceito de intermédio entendido como âmbito ou espaço intersticial, fronteiriço e ambivalente (ou multivalente), atravessa na presente investigação o campo da etimologia, da ciência, do pensamento e da arte, para deter-se na arquitectura actual, realizando-se uma indagação concreta no processo de concepção, na proposta formal, espacial e funcional, assim como na percepção e demais experiencias ou vivencias no lugar arquitectónico. 3. Em primeiro lugar, explora-se o conceito de “intermédio” nas suas características, funções e concretizações na cultura contemporânea, detectando os seus antecedentes culturais mais importantes, para em seguida aplicá-lo reflexivamente a obras significativas de arquitectura. A investigação centra-se na modalidade figural (imprecisa) do intermédio (alusão ao figural deleuziano) cujas metáforas base são o translúcido, a penumbra e a espuma, relacionadas com o actual paradigma da complexidade (C. Jencks). Em seguida analisam-se as relações ou implicações da referida modalidade figural em obras de arquitectos como Eisenman, Holl, Tschumi, Ito, Fujimoto, Van Berkel e Bos, e Siza, como modo de comprovar o seu valor operativo e revelar o seu sentido e importância na actualidade e na disciplina. O estudo tem como referencia o conceito de in-between da obra de Eisenman, e utiliza conceitos da filosofia do limite de Trias como alguns dos seus principais fundamentos. 4. Em cada uma das obras de arquitectura apresentadas esta modalidade imprecisa traduz-se num âmbito ou espaço intermédia figural especifico. São obras nas quais real e virtual, matricial e objectual, arquitectura e território, existente e novo, colectivo e individual, social e intimo, interior e exterior, e outros usos e funções se entrelaçam ou fundem. São arquitecturas cuja geometria já não está tão determinada por aspectos bi ou tridimensionais do desenho ou da construção. Através do uso destas geometrias mais ou menos complexas, dinâmicas e intersticiais, estas obras reflectem (ou intuem) as premissas do actual zeitgeist: uma mudança de esquemas de objectos a relações para um pensamento mais holístico, transdisciplinar, sistémico ou complexo (E. Morin) e uma nova consciência colectiva sobre a realidade, que anuncia a crise da percepção, a mudança de paradigma e novos valores (F. Capra). Emerge uma outra forma de sentir e perceber o mundo, os lugares e os espaços, que gradualmente vai alterando o modo de pensar e desenhar a arquitectura, e consequentemente de interagir com ela. 5. O sentido do espaço intermédio figural, emerge nas cada vez mais complexas qualidades morfológico-espaciais e funcionais de uma parte importante da arquitectura actual. Cada vez mais, arquitectura, geometria e tecnologia informática, relacionam-se de um modo mais livre para indagar outras formas de pensar e fazer lugares, onde os espaços intermédios que investigamos são um testemunho. O espaço arquitectónico torna-se afectivo e interactivo, um lugar intermédio figural, formalizado por espacialidades intersticiais que parecem aludir ao estado liquido, onde a forma é mais matriz que configuração, o espaço mais espacialidade que compartimento, a função mais versátil e multivalente que especifica ou autónoma, e a perplexidade, a imaginação e a evocação entrecruzam-se e tornam-se patentes. 6. A arquitectura dos lugares intermédios valoriza não só o “plano”, a “configuração” e a “transparência”, mas sobretudo o” espaço” e a ”relação” nas suas inter-relações com os utentes e restante contextos. Da identidade à relacionalidade, da representação à apresentação, é uma arquitectura que propõe “outros espaços”, que além de espaços são topografias e espacialidades intersticiais, de tensão, transição, transformação, relação, intercambio e interacção. São lugares intermédios reais e virtuais que utilizam tanto a morfologia como a topologia para conquistar novas espacialidades, pretendendo ultrapassar a estrita operação de “forming”/“morphing”/formação para explorar a de “spacing”/espaçamento. São lugares que se apoiam tanto no conceito de zona de indescernibilidade de Deleuze, como na de imagem-fluxo de Buci-Glucksmann como no de suspensão–entre de Sloterdijk ou de espaço-não objectivo que refere José Gil, e se plasmam na arquitectura contemporânea como tropos do intersticial. 7. São os lugares intermédios da arquitectura actual, de espacialidade háptica, mais “afectiva”, generativa e interactiva, onde esse âmbito intermédio transforma e é transformado, “afecta”, ”co-move “(nos faz mover/agir) e evoca outros lugares, outras possibilidades de espaço habitável ou ocupável. Emerge o intermédio como lugar que, paradigmática e manifestamente, o monumento de Eisenmann em Berlim, anuncia e exibe.

El tiempo como sustancia de la forma : una aproximación al Museo de Arte Romano de Mérida desde los presupuestos del vitalismo

El modelo dominante durante la Era Moderna asume la presencia del hombre como sujeto dentro del gran engranaje mecánico del Cosmos. Asimismo, recoge una idea acerca del ser natural dentro de la tradición ontológica iniciada por el eleatismo presocrático que concibe a éste como lo inmutable y estático frente al cambio y al movimiento, los cuales se constituyen como meras apariencias. Durante el periodo anterior a la aparición de los grandes filósofos griegos se produce una transformación en donde, de la inicial cosmología vinculada a un tiempo primordial, se pasará a una visión del Universo como ente indestructible, atemporal, inmutable, perfecto, geométrico y espacial. Si en Demócrito se admite un universo sometido al azar y a la necesidad, en Platón el Universo sólo atiende a la necesidad. Este modelo ontológico se pone en entredicho cuando el hombre ya no es concebido como pieza de un sistema más amplio, sino como centro radical del pensamiento. La condición radical del hombre es entonces su propia vida, siendo éste el concepto troncal del denominado vitalismo cuyo más influyente representante en España es José Ortega y Gasset. El estatismo del ser –del hombre- pierde sentido; en palabras del propio Ortega, “no es un ser sino un estar siendo” lo que caracteriza a la vida humana. La razón cartesiana es ahora la razón vital y su objeto de estudio no es la naturaleza sino el propio devenir, es decir, el tiempo, la historia. Este planteamiento es fundamental para comprender el edificio que es objeto de este estudio, el Museo de Arte Romano de Mérida (1980-1985) de Rafael Moneo. Por ello el concepto de tiempo es utilizado como marco y estructura de la presente tesis, a sabiendas del notorio y muy significativo papel que este edificio desempeñó en la carrera de su autor y en el panorama nacional e internacional de la arquitectura y de la museología. Este proyecto nos permite acercarnos al pensamiento de su autor a través de un edificio que, aun habiendo sido ampliamente reconocido, no cuenta con un estudio suficientemente exhaustivo que recoja la amplitud y riqueza que encierra. Esta tesis no es un compendio de lo que ya se ha investigado sobre Mérida; es una aproximación global e interpretativa cuyo sentido sólo puede concebirse al vislumbrar la estructura completa de la misma en sintonía con el "lógos" vital, histórico y narrativo que el proyecto encierra. Se revisará la concepción histórica según la cual, la configuración espacial de la forma habría tenido primacía respecto a su configuración temporal, al remitir esta última a una condición espacializada y circunstancial. La componente vicaria de la circunstancia será elevada por Ortega a la categoría de esencial, visualizando así una paradoja cuya reformulación nos lleva a la concepción de un tiempo sustancial. El diccionario de la RAE, en su tercera acepción, define la sustancia como “aquello que permanece en algo que cambia”, lo cual nos remite al pensamiento antiguo. Se mostrará que lo que permanece no necesariamente implica una concepción estática y eleática de la forma, que la arquitectura esencial no es unívocamente la arquitectura atemporal del platonismo y que cabe concebir la "firmitas" desde la atención a la "durée" bergsoniana. Al asociar tradicionalmente la sustancia con el referido estatismo, se margina al tiempo y a la duración a lo no sustancial; por ello, se tratará de aproximar los términos de tiempo y sustancia para definir la forma. Ello conllevará al estudio de las notables patologías derivadas de la asunción de un tiempo cronológico en nuestra contemporaneidad frente a las cuales, las intuiciones contenidas en Mérida, se alinearán con la actual actitud revisionista en el ámbito del pensamiento filosófico y científico. En Mérida, la memoria recogerá los aspectos de la conciencia así como los aspectos vinculados a la experiencia íntima y colectiva como soporte para la consecución de un discurso. La dualidad entre intuición e inteligencia será recogida por Moneo con idea de trascender su incomunicabilidad, mediante una operación que consistirá en la reivindicación de una memoria irreductible, cuya morada estaría incardinada en el propio tiempo de la duración y de la vida y no en la espacialidad coextensiva del presente y de la acción funcional sobre la materia. Moneo asumirá el papel de la memoria como condición central de una forma que se encarnará al concebirse como un teatro. En la respuesta a la contradicción entre el hecho físico y el efecto psíquico de la experiencia humana residirá la pertinencia de un tiempo narrativo. Será entonces el lenguaje el encargado de aportar sentido a la obra mediante el recurso fundado en la dramatización de la experiencia, es decir, a través de una conexión entre la conciencia íntima y el carácter social y colectivo intrínseco en la arquitectura. ABSTRACT TIME AS A SUBSTANCE OF FORM. AN APPROACH TO THE ROMAN ART MUSEUM OF MÉRIDA FROM THE VIEWPOINT OF VITALISM. The dominant model during the Modern Era placed man as a subject inside the great mechanism of the cosmos. It is also based in an idea about natural being within the ontological tradition initiated by the pre-Socratic Eleatism that conceives it as something immutable and static in respect with change and movement, which are considered as mere appearances. Prior to the emergence of the great Greek philosophers occurred a transformation where concepts of cosmology linked to a primordial time, changed to a view of the universe as indestructible, timeless, unchanging, perfect, geometric and spatial. If Democritus accepted a universe subjected to randomness and necessity, Plato thought that the universe only worked by necessity. This ontological model is called into question when man is not conceived as a piece of a broader system, but as a radical center of thinking. The radical condition of man then is his own life. This is the core concept of so-called Vitalism, whose most influential representative in Spain was José Ortega y Gasset. The stillness of being – of man - loses its meaning; in the words of Ortega, “it is not being but being in progress” that characterizes human life. The Cartesian reason is now the vital reason and its subject of study is no longer nature but its own evolution, in other words, time and history. This approach is fundamental to understand the building which is the subject of this study, the Museum of Roman Art in Mérida (1980-1985) by Rafael Moneo. The concept of time is used as a framework and structure of this thesis, demonstrating the notorious and very significant role this building has implied in the career of its author and in the national and international panorama of architecture and museology. This project allow us to approach the thought of its author through a building that, even whilst widely recognized, does not yet have a sufficiently comprehensive study covering its breadth and richness. This thesis is not a compendium of what already has been researched on Merida; it is a global and interpretative approach whose meaning can only be conceived as a study of its complete structure in line with the vital, historical and narrative logos the project implies. We will review the historical idea where spatial configuration of the form would have had primacy with respect to temporary configuration, because the latter refers to a spatial and circumstantial condition. The vicarious nature of the circumstance will be elevated by Ortega to the category of essential, thus showing a paradox which reformulation leads us to the conception of a substantial time. The dictionary of the Spanish Royal Academy, in its third meaning, defines substance as "that which remains in something that changes". This is a reference to ancient thought. It will be shown that what remains does not necessarily imply a static and Eleatic conception of form. It will also be shown that the essential architecture is not uniquely the timeless architecture of Platonism and that it is possible to conceive the "firmitas" parallel to the "durée" of Henri Bergson. As a result of this traditional association between substance and stillness, it marginalizes the time and the duration to the non-substantial; for this reason, we will try to approach terms of time and substance to define the shape. This will involve studying significant pathologies resulting from an assumption of chronological time in our contemporary world against which, the insights contained in Merida, will be aligned with the current revisionist attitude in the fields of philosophical and scientific thought. In Merida, memory includes aspects of consciousness as well as aspects linked to the intimate and collective experience as a foundation for the achievement of discourse. The duality between intuition and intelligence is put forward by Moneo with the idea of transcending its lack of communication, by means of a resource consisting of the vindication of an irreducible memory, whose home would be embodied in the time of duration and life and not in the coextensive spatiality of the present and in the functional action on the matter. Moneo demonstrates the role of memory as a central condition of form as a theatre. In response to the contradiction between the physical fact and the psychological effect of human experience lies the relevance of narrative time. Language then assumes the responsibility of giving meaning to the work through the dramatization of experience, i.e., through a connection between the intimate consciousness and the intrinsic social and collective character of architecture.

A DIALÉTICA ENTRE O CONHECIMENTO DE SI E O CONHECIMENTO DE DEUS NO LIVRO X DAS CONFISSÕES DE SANTO AGOSTINHO

O conhecimento de si e o conhecimento de Deus estabelecem uma relação fundamental na obra mais conhecida de Agostinho, Confissões. O livro X das Confissões contém as narrativas centrais para a análise da dialética entre o conhecimento de si e o conhecimento de Deus, que tem como chave de leitura a memória para a constituição do cogito existencial. É examinada a relação que existe, no texto narrativo de Agostinho, entre a interpretação da Escritura e a constituição do si, em que há aspectos do discurso interior e abordagem no quadro da teoria narrativa que é dada a partir do conceito de identidade narrativa. A constituição do si é desenvolvida na dialética interna do personagem entre a afirmação de si e a negação de si, que apresenta a imanência do homem como característica pessoal e, ao mesmo tempo, o desejo de transcendência daquilo que o ser humano tem de mais íntimo em relação a Deus. Esta é uma análise da tensão existente entre a visão que o ser humano tem da consciência de si e a que ele tem de Deus, na busca pela felicidade.(AU)

A DIALÉTICA ENTRE O CONHECIMENTO DE SI E O CONHECIMENTO DE DEUS NO LIVRO X DAS CONFISSÕES DE SANTO AGOSTINHO

O conhecimento de si e o conhecimento de Deus estabelecem uma relação fundamental na obra mais conhecida de Agostinho, Confissões. O livro X das Confissões contém as narrativas centrais para a análise da dialética entre o conhecimento de si e o conhecimento de Deus, que tem como chave de leitura a memória para a constituição do cogito existencial. É examinada a relação que existe, no texto narrativo de Agostinho, entre a interpretação da Escritura e a constituição do si, em que há aspectos do discurso interior e abordagem no quadro da teoria narrativa que é dada a partir do conceito de identidade narrativa. A constituição do si é desenvolvida na dialética interna do personagem entre a afirmação de si e a negação de si, que apresenta a imanência do homem como característica pessoal e, ao mesmo tempo, o desejo de transcendência daquilo que o ser humano tem de mais íntimo em relação a Deus. Esta é uma análise da tensão existente entre a visão que o ser humano tem da consciência de si e a que ele tem de Deus, na busca pela felicidade.(AU)

HOP-1, a Caenorhabditis elegans presenilin, appears to be functionally redundant with SEL-12 presenilin and to facilitate LIN-12 and GLP-1 signaling

Mutant presenilins have been found to cause Alzheimer disease. Here, we describe the identification and characterization of HOP-1, a Caenorhabditis elegans presenilin that displays much more lower sequence identity with human presenilins than does the other C. elegans presenilin, SEL-12. Despite considerable divergence, HOP-1 appears to be a bona fide presenilin, because HOP-1 can rescue the egg-laying defect caused by mutations in sel-12 when hop-1 is expressed under the control of sel-12 regulatory sequences. HOP-1 also has the essential topological characteristics of the other presenilins. Reducing hop-1 activity in a sel-12 mutant background causes synthetic lethality and terminal phenotypes associated with reducing the function of the C. elegans lin-12 and glp-1 genes. These observations suggest that hop-1 is functionally redundant with sel-12 and underscore the intimate connection between presenilin activity and LIN-12/Notch activity inferred from genetic studies in C. elegans and mammals.

The circadian clock controls the expression pattern of the circadian input photoreceptor, phytochrome B

Developmental and physiological responses are regulated by light throughout the entire life cycle of higher plants. To sense changes in the light environment, plants have developed various photoreceptors, including the red/far-red light-absorbing phytochromes and blue light-absorbing cryptochromes. A wide variety of physiological responses, including most light responses, also are modulated by circadian rhythms that are generated by an endogenous oscillator, the circadian clock. To provide information on local time, circadian clocks are synchronized and entrained by environmental time cues, of which light is among the most important. Light-driven entrainment of the Arabidopsis circadian clock has been shown to be mediated by phytochrome A (phyA), phytochrome B (phyB), and cryptochromes 1 and 2, thus affirming the roles of these photoreceptors as input regulators to the plant circadian clock. Here we show that the expression of PHYB∷LUC reporter genes containing the promoter and 5′ untranslated region of the tobacco NtPHYB1 or Arabidopsis AtPHYB genes fused to the luciferase (LUC) gene exhibit robust circadian oscillations in transgenic plants. We demonstrate that the abundance of PHYB RNA retains this circadian regulation and use a PHYB∷Luc fusion protein to show that the rate of PHYB synthesis is also rhythmic. The abundance of bulk PHYB protein, however, exhibits only weak circadian rhythmicity, if any. These data suggest that photoreceptor gene expression patterns may be significant in the daily regulation of plant physiology and indicate an unexpectedly intimate relationship between the components of the input pathway and the putative circadian clock mechanism in higher plants.

MCD4 Encodes a Conserved Endoplasmic Reticulum Membrane Protein Essential for Glycosylphosphatidylinositol Anchor Synthesis in Yeast

Glycosylphosphatidylinositol (GPI)-anchored proteins are cell surface-localized proteins that serve many important cellular functions. The pathway mediating synthesis and attachment of the GPI anchor to these proteins in eukaryotic cells is complex, highly conserved, and plays a critical role in the proper targeting, transport, and function of all GPI-anchored protein family members. In this article, we demonstrate that MCD4, an essential gene that was initially identified in a genetic screen to isolate Saccharomyces cerevisiae mutants defective for bud emergence, encodes a previously unidentified component of the GPI anchor synthesis pathway. Mcd4p is a multimembrane-spanning protein that localizes to the endoplasmic reticulum (ER) and contains a large NH2-terminal ER lumenal domain. We have also cloned the human MCD4 gene and found that Mcd4p is both highly conserved throughout eukaryotes and has two yeast homologues. Mcd4p’s lumenal domain contains three conserved motifs found in mammalian phosphodiesterases and nucleotide pyrophosphases; notably, the temperature-conditional MCD4 allele used for our studies (mcd4–174) harbors a single amino acid change in motif 2. The mcd4–174 mutant (1) is defective in ER-to-Golgi transport of GPI-anchored proteins (i.e., Gas1p) while other proteins (i.e., CPY) are unaffected; (2) secretes and releases (potentially up-regulated cell wall) proteins into the medium, suggesting a defect in cell wall integrity; and (3) exhibits marked morphological defects, most notably the accumulation of distorted, ER- and vesicle-like membranes. mcd4–174 cells synthesize all classes of inositolphosphoceramides, indicating that the GPI protein transport block is not due to deficient ceramide synthesis. However, mcd4–174 cells have a severe defect in incorporation of [3H]inositol into proteins and accumulate several previously uncharacterized [3H]inositol-labeled lipids whose properties are consistent with their being GPI precursors. Together, these studies demonstrate that MCD4 encodes a new, conserved component of the GPI anchor synthesis pathway and highlight the intimate connections between GPI anchoring, bud emergence, cell wall function, and feedback mechanisms likely to be involved in regulating each of these essential processes. A putative role for Mcd4p as participating in the modification of GPI anchors with side chain phosphoethanolamine is also discussed.

Accumulation of Major Histocompatibility Complex Class II Molecules in Mast Cell Secretory Granules and Their Release upon Degranulation

To investigate the relationship between major histocompatibility complex (MHC) class II compartments, secretory granules, and secretory lysosomes, we analyzed the localization and fate of MHC class II molecules in mast cells. In bone marrow-derived mast cells, the bulk of MHC class II molecules is contained in two distinct compartments, with features of both lysosomal compartments and secretory granules defined by their protein content and their accessibility to endocytic tracers. Type I granules display internal membrane vesicles and are accessed by exogenous molecules after a time lag of 20 min; type II granules are reached by the endocytic tracer later and possess a serotonin-rich electron-dense core surrounded by a multivesicular domain. In these type I and type II granules, MHC class II molecules, mannose-6-phosphate receptors and lysosomal membrane proteins (lamp1 and lamp2) localize to small intralumenal vesicles. These 60–80-nm vesicles are released along with inflammatory mediators during mast cell degranulation triggered by IgE-antigen complexes. These observations emphasize the intimate connection between the endocytic and secretory pathways in cells of the hematopoietic lineage which allows regulated secretion of the contents of secretory lysosomes, including membrane proteins associated with small vesicles.

The Dynamin-like Protein DLP1 Is Essential for Normal Distribution and Morphology of the Endoplasmic Reticulum and Mitochondria in Mammalian Cells

The dynamin family of large GTPases has been implicated in vesicle formation from both the plasma membrane and various intracellular membrane compartments. The dynamin-like protein DLP1, recently identified in mammalian tissues, has been shown to be more closely related to the yeast dynamin proteins Vps1p and Dnm1p (42%) than to the mammalian dynamins (37%). Furthermore, DLP1 has been shown to associate with punctate vesicles that are in intimate contact with microtubules and the endoplasmic reticulum (ER) in mammalian cells. To define the function of DLP1, we have transiently expressed both wild-type and two mutant DLP1 proteins, tagged with green fluorescent protein, in cultured mammalian cells. Point mutations in the GTP-binding domain of DLP1 (K38A and D231N) dramatically changed its intracellular distribution from punctate vesicular structures to either an aggregated or a diffuse pattern. Strikingly, cells expressing DLP1 mutants or microinjected with DLP1 antibodies showed a marked reduction in ER fluorescence and a significant aggregation and tubulation of mitochondria by immunofluorescence microscopy. Consistent with these observations, electron microscopy of DLP1 mutant cells revealed a striking and quantitative change in the distribution and morphology of mitochondria and the ER. These data support very recent studies by other authors implicating DLP1 in the maintenance of mitochondrial morphology in both yeast and mammalian cells. Furthermore, this study provides the first evidence that a dynamin family member participates in the maintenance and distribution of the ER. How DLP1 might participate in the biogenesis of two presumably distinct organelle systems is discussed.

Transcription factor Mts1/Mts2 (Atf1/Pcr1, Gad7/Pcr1) activates the M26 meiotic recombination hotspot in Schizosaccharomyces pombe

Homologous recombination hotspots increase the frequency of recombination in nearby DNA. The M26 hotspot in the ade6 gene of Schizosaccharomyces pombe is a meiotic hotspot with a discrete, cis-acting nucleotide sequence (5′-ATGACGT-3′) defined by extensive mutagenesis. A heterodimeric M26 DNA binding protein, composed of subunits Mts1 and Mts2, has been identified and purified 40,000-fold. Cloning, disruption, and genetic analyses of the mts genes demonstrate that the Mts1/Mts2 heterodimer is essential for hotspot activity. This provides direct evidence that a specific trans-acting factor, binding to a cis-acting site with a unique nucleotide sequence, is required to activate this meiotic hotspot. Intriguingly, the Mts1/Mts2 protein subunits are identical to the recently described transcription factors Atf1 (Gad7) and Pcr1, which are required for a variety of stress responses. However, we report differential dependence on the Mts proteins for hotspot activation and stress response, suggesting that these proteins are multifunctional and have distinct activities. Furthermore, ade6 mRNA levels are equivalent in hotspot and nonhotspot meioses and do not change in mts mutants, indicating that hotspot activation is not a consequence of elevated transcription levels. These findings suggest an intimate but separable link between the regulation of transcription and meiotic recombination. Other studies have recently shown that the Mts1/Mts2 protein and M26 sites are involved in meiotic recombination elsewhere in the S. pombe genome, suggesting that these factors help regulate the timing and distribution of homologous recombination.

Physical and functional association of glycolipid N-acetyl-galactosaminyl and galactosyl transferases in the Golgi apparatus

Glycolipid glycosyltransferases catalyze the stepwise transfer of monosaccharides from sugar nucleotides to proper glycolipid acceptors. They are Golgi resident proteins that colocalize functionally in the organelle, but their intimate relationships are not known. Here, we show that the sequentially acting UDP-GalNAc:lactosylceramide/GM3/GD3 β-1,4-N-acetyl-galactosaminyltransferase and the UDP-Gal:GA2/GM2/GD2 β-1,3-galactosyltransferase associate physically in the distal Golgi. Immunoprecipitation of the respective epitope-tagged versions expressed in transfected CHO-K1 cells resulted in their mutual coimmunoprecipitation. The immunocomplexes efficiently catalyze the two transfer steps leading to the synthesis of GM1 from exogenous GM3 in the presence of UDP-GalNAc and UDP-Gal. The N-terminal domains (cytosolic tail, transmembrane domain, and few amino acids of the stem region) of both enzymes are involved in the interaction because (i) they reproduce the coimmunoprecipitation behavior of the full-length enzymes, (ii) they compete with the full-length counterpart in both coimmunoprecipitation and GM1 synthesis experiments, and (iii) fused to the cyan and yellow fluorescent proteins, they localize these proteins to the Golgi membranes in an association close enough as to allow fluorescence resonance energy transfer between them. We suggest that these associations may improve the efficiency of glycolipid synthesis by channeling the intermediates from the position of product to the position of acceptor along the transfer steps.

Exploitation of host cells by enteropathogenic Escherichia coli

Microbial pathogens have evolved many ingenious ways to infect their hosts and cause disease, including the subversion and exploitation of target host cells. One such subversive microbe is enteropathogenic Escherichia coli (EPEC). A major cause of infantile diarrhea in developing countries, EPEC poses a significant health threat to children worldwide. Central to EPEC-mediated disease is its colonization of the intestinal epithelium. After initial adherence, EPEC causes the localized effacement of microvilli and intimately attaches to the host cell surface, forming characteristic attaching and effacing (A/E) lesions. Considered the prototype for a family of A/E lesion-causing bacteria, recent in vitro studies of EPEC have revolutionized our understanding of how these pathogens infect their hosts and cause disease. Intimate attachment requires the type III-mediated secretion of bacterial proteins, several of which are translocated directly into the infected cell, including the bacteria's own receptor (Tir). Binding to this membrane-bound, pathogen-derived protein permits EPEC to intimately attach to mammalian cells. The translocated EPEC proteins also activate signaling pathways within the underlying cell, causing the reorganization of the host actin cytoskeleton and the formation of pedestal-like structures beneath the adherent bacteria. This review explores what is known about EPEC's subversion of mammalian cell functions and how this knowledge has provided novel insights into bacterial pathogenesis and microbe-host interactions. Future studies of A/E pathogens in animal models should provide further insights into how EPEC exploits not only epithelial cells but other host cells, including those of the immune system, to cause diarrheal disease.