Decodificar Sejima‐SANAA: Estrategias formales y estructurales en su trayectoria de proyecto, 1987 a 2010; de la Casa Platform I al Centro Rolex

La Tesis decodifica una selección de veinte proyectos representativos de Sejima-SANAA, desde su primer proyecto construido, la Casa Platform I, 1987, hasta el Centro Rolex, 2010, año en que Sejima y Nishizawa –SANAA- reciben el Premio Pritzker. De los veinte proyectos once son de Sejima: Casa Platform I, Casa Platform II, Residencia de Mujeres, Casa N, Pachinco Parlor I, Villa en el Bosque, Comisaría en Chofu, Casa Y, Apartamentos en Gifu, Edificio de equipamientos en la Expo Tokio 96, Pachinko Parlor III; y nueve de SANAA: edificio Multimedia en Oogaki, estudio de viviendas metropolitanas,Park Café en Koga, De Kunstlinie en Almere, Museo de Kanazawa, Pabellón de Toledo, Escuela de Zollverein, Casa Flor y Centro Rolex. La decodificación lee la obra de Sejima-SANAA a la inversa para ‘reconstruir’, en un ejercicio de simulación ficticia, una versión verosímil y coherente de los que podrían haber sido sus procesos proyectuales; podrían, porque los verdaderos son imposibles de dilucidar. Los que se proponen se pretenden exclusivamente verosímiles y plausibles. Con ello se pretende contribuir al entendimiento y comprensión de la arquitectura de Sejima-SANAA y, tangencialmente y en menor medida, a la teoría sobre el ejercicio proyectual arquitectónico. La decodificación se centra en dos aspectos concretos: la forma arquitectónica y el papel proyectual de la estructura portante. Ambas decodificaciones se extienden inevitablemente a otros aspectos relacionados, como, por ejemplo, la naturaleza del espacio arquitectónico. El procedimiento de investigación partió de una descripción objetiva y pormenorizada de los significantes formales y estructurales de cada proyecto desde su propia configuración física y geométrica. Esa descripción ‘objetiva’, llevada al límite, permitió que afloraran estructuras conceptuales y lógicas subyacentes de cada proyecto. Unida a interpretación crítica, –mediante su relación y confrontación con otras arquitecturas y otros modos de hacer conocidos- permitió trazar la reconstitución ficticia que persigue la decodificación. Ese trabajo se materializó en veinte ensayos críticos y se acompañó de un conjunto de otros textos sobre temas sugeridos o reclamados por el proceso de investigación. El conjunto de todos esos textos constituye el material de trabajo de la tesis. A partir de ahí, con una visión de conjunto, la tesis identifica una trayectoria de estrategias formales y una trayectoria de estrategias proyectuales relacionadas con lo portante. Juntas conforman el grueso de la tesis que se expone en los cuatro capítulos centrales. Los precede un capítulo introductorio que expone el recorrido biográfico de K. Sejima y la trayectoria profesional de Sejima-SANAA; y los siguen de unos textos transversales sobre forma, lugar y espacio. La tesis termina con una síntesis de sus conclusiones. Las estrategias formales se exponen en tres capítulos. El primero, ‘Primeras estrategias formales’ agrupa proyectos de la primera etapa de Sejima. El segundo capítulo está dedicado enteramente al proyecto de los apartamentos en Gifu, 1994-98, que según esta tesis, supuso un importante punto de inflexión en la trayectoria de Sejima; tanto el tercer capítulo lleva por nombre ‘Estrategias formales después de Gifu’ y recoge los proyectos que le siguieron. Las ‘Primeras estrategias formales’, varias y balbucientes, se mueven en general en torno a dos modos o procedimientos de composición, bien conocidos: por partes y sistemático. Éste última inicia en la trayectoria de SANAA un aspecto que va a ser relevante de aquí en adelante: entender el proyecto como propuesta genérica en la que, más allá de su realidad específica y tangible, subyace una lógica, en cada proyecto la suya, extrapolable a otros lugares, otras dimensiones, incluso otros programas: cada proyecto podría dar lugar a otros proyectos de la misma familia. La composición sistemática incluye, entre otros, la Casa Platform II, basada en la definición de un elemento constructivo, y la formulación de unas leyes de repetición y de posibles modos de agrupación. Incluye también la Residencia de Mujeres Saishunkan Seiyaku- proyecto que lanzó a Sejima a la fama internacional-, que también sería un sistema, pero distinto: basado en la repetición regular de una serie de elementos a lo largo de una directriz generando un hipotético contenedor infinito del que el proyecto sería tan solo un fragmento. La estrategia formal del edificio de Gifu ahondaría en la voluntad genérica del proyecto, adoptando la lógica de un juego. El proyecto sería una partida del juego, pero no la única posible, podrían jugarse otras. Esta hipótesis del juego está verificada en ‘El Juego de Gifu’ que - tras formular el juego identificando sus elementos (tablero y fichas), reglas y procedimientos- juega una partida: la que habría dado lugar al edificio proyectado por Sejima. Gifu extiende el concepto de ‘repetir’ un elemento constructivo a la de repetir un patrón espacial, lo que conlleva: la desvinculación entre forma y función; y un nuevo concepto de flexibilidad, que deja de referirse al uso flexible del edificio construido para pertenecer al momento proyectual en que se asignan funciones específicas a los patrones espaciales. Esta tesis propone que esa asignación de funciones sería uno de los últimos eslabones del proceso proyectual, algo opuesto a la premisa moderna de “la forma sigue a la función”. Las estrategias formales ‘Después de Gifu’ tienen también lógicas de juego, pero cada estrategia responde a un juego distinto, como dejan entrever sus nombres: ‘Tableros de Juego’, que con distintos grados de madurez estaría presente en varios proyectos; ‘Elementos de Catálogo’ en el Museo de Kanazawa; ‘Forma apriorística’, en la Casa Flor y ‘Repetición de una situación topológica’, en el Centro Rolex. Todas esas estrategias, o juegos, mantienen aspectos comunes relativos a la forma arquitectónica, precisamente los aspectos Gifu: la repetición aplicada al patrón espacial, y lo que conlleva: desvinculación entre forma y función y la nueva acepción de flexibilidad. ‘Tableros de Juego’ consiste en configurar cada sistema de proyecto (estructura, cerramientos, particiones y mobiliario) eligiendo elementos ofrecidos por una geometría de base, en cada proyecto la suya, en general reticular: intersecciones, líneas, módulos. Cada sistema se configura, en principio, sin relación de subordinación con cualquiera de los demás; cuando esa subordinación es ineludible, el juego determina que el sistema portante no puede materializar el orden geométrico de base, lo que se traduce en que no ejerce el papel dominante. Por lo tanto, ‘Tableros de Juego’ transgrede la lógica de la planta libre moderna: la estructura ni refleja ni revela el orden de base y los sistemas no respetan las relaciones de subordinación jerárquica y encadenada que aquella determinaba. Esta estrategia de ‘Tableros de juego’ deriva en soluciones y proyectos formales muy distintos: los proyectos de Oogaki y Park Café, que presentarían ‘Tableros de Juego’ incipientes; De Kunstlinie en Almere y la Escuela de Zollverein, que presentarían una consolidación de esta estrategia; y el Pabellón de Vidrio de Toledo que resultaría de la subversión de la estrategia. Este último proyecto, además, lleva el concepto de repetición más allá del elemento constructivo y del patrón espacial (que en este caso tiene forma de burbuja) parar acabar afectando a la propia experiencia del espectador, que esté donde esté, siempre tiene la sensación de estar en el mismo sitio. Esta tesis denomina a ese espacio repetitivo como ‘espacio mantra’. La estrategia ‘Elementos de Catálogo’ se ilustra con el Museo de Kanazawa. Su lógica parte de la definición de una serie de elementos, muy pocos, y se basa en el ingente número de posibles combinaciones entre sí. Gifu habría anunciado el catalogo de elementos en la caracterización de sus patrones espaciales. La estrategia ‘Forma Apriorística’ se ilustra con la Casa Flor. La decisión sobre el tipo de forma -en este caso la de una ameba- estaría al principio del proceso proyectual, lo que no quiere decir que sea una forma arbitraria: la forma de la ameba lleva implícita la repetición de un patrón espacial (el seudópodo) y una apoteosis del concepto de repetición que, alcanzando la experiencia espacial, da lugar a un espacio repetitivo o mantra. El ‘Espacio Mantra’ es uno de los leitmotivs, que se emplean como argumento en la última estrategia formal que la Tesis decodifica: el Centro Rolex. Con respecto a la estructura portante, la tesis identifica y traza una trayectoria de cinco estrategias proyectuales: preeminencia, ocultación, disolución, desaparición y desvirtuación. --Ocultación, reduce el papel dominante de la estructura. Al principio es una ocultación literal, casi un tapado de los elementos estructurales, como en Gifu; luego se hace más sofisticada, como la ocultación por camuflaje o la paradójica ocultación por multiplicación de Park Café. --La disolución merma la condición dominante de la estructura que en lugar de configurarse como sistema unitario u homogéneo se fragmenta en varios subsistemas. --La desaparición se refiere a estructuras que desaparecen como sistemas propios y autónomos, a proyectos en los que la función portante es desempeñada por otros sistemas como el de las particiones. La desaparición culmina con la Casa Flor, cuyo perímetro ejerce la función portante y además es transparente, está desmaterializado: la estructura se ha hecho invisible, ha desaparecido. --La desvirtuación se refiere a estructuras que sí se presentan como sistemas propios y autónomos, pero dejan de tener un papel preeminente por cuanto no materializan el orden de base: esta estrategia es correlativa a la estrategia formal ‘Tableros de juego’. Las conclusiones de la tesis están en la propia organización de la tesis: la identificación de las estrategias. Aún así, y como epílogos, se exponen seis. Las dos primeras subrayan el hilo conductor del trabajo realizado, que radica en la cualidad genérica de las estrategias proyectuales en Sejima-SANAA. Las cuatro siguientes dilucidan hasta qué punto hay, en sus proyectos, rasgos o significantes formales y/o estructurales que sean a su vez señales características del panorama arquitectónico contemporáneo; y plantean la pregunta estrella: ¿hay algunos que, apuntando más lejos, supongan aportaciones originales? --Como aportaciones originales la tesis destaca: la identificación entre el ideal genérico y proyecto concreto; y la propuesta de un espacio nuevo, híbrido, una suerte de estadio intermedio entre el espacio subdividido y compartimentado de la tradición y el continuo moderno. --Como síntomas de contemporaneidad se destacan: respecto de la forma, la traslación de la especificidad formal de la parte al conjunto; y respecto de la estructura, la tendencia contemporánea a hacer estructuras cada vez más ligeras y livianas, que tienden a lo evanescente. Ésta última, la tendencia al evanescencia estructural, podría tener la condición de aportación original, no en vano la desaparición de la estructura lleva la evanescencia hacia sus últimas consecuencias, y en el caso de estructuras con presencia física, hace que dejen de ser el sistema ordenador orquestador del proceso proyectual. ABSTRACT The Thesis decodes a selection of twenty representative Sejima-SANAA projects, from the first one built, the Platform I House in 1987, to the Rolex Center in 2010, year in which Sejima and Nishizawa –SANAA- received the Pritzker Prize. Eleven projects are from Sejima: Platform I, Platform II, Saishunkan Seiyaku Women´s Dormitory, N- House, Pachinco Parlor I, Villa in the Forest, Policy Box at Chofu Station, Y-House, Gifu Kitigata Apartment, World City Expo ´96 Facilities Building, Pachinko Parlor III; and nine from SANAA: Multimedia Workshop in Ogaki, Metropolitan Housing Studies, Park Café in Koga, De Kunstlinie in Almere, Kanazawa Museum, Glass Pavilion at the Toledo Museum of Art, Zollverein School, Flower House and the Rolex Center. This decoding reads the Sejima-SANAA’s projects inversely aiming ‘to reconstruct', in a fictitious simulation exercise, a likely and coherent version of what her/their projectual processes ‘could’ have been; ‘could’, because the true ones are impossible to explain. The ones proposed here pretend only to be likely and reasonable. By so doing the Thesis tries to contribute to the understanding and comprehension of Sejima-SANAA architecture and, tangentially and to a lesser extent, to the theory of architectural projects exercise. Decoding centers in two specific aspects: architectural form, and projectual role of the load bearing structure. Both decodes inevitably extend to other related aspects such as, for example, the nature of space. The research procedure begun by carrying out an objective and detailed description of the formal and structural signifiers of each project; looking at them from their physical and geometric configuration. Taken to the limit, the ‘objective’ descriptions allowed the conceptual structures and underlying logics of each project to arise. Together with critical interpretations, which related and confronted them with other architectures and well-known projectual working ways, it became possible to outline and trace the intended fictitious reconstruction decodes. The descriptive analytical work materialized in twenty critical essays, and was accompanied by a set of other essays on subjects suggested or demanded by the research process. Together, all those texts were the material basis on which thesis work was built. Looking at the whole and taking it from there, the thesis identifies two related projectual trajectories: a trajectory of formal strategies and a trajectory of strategies having to do with structural systems and components. Both, together, constitute the bulk of the thesis, as presented in the four central chapters. Preceding them there is an introductory chapter outlining the biographical path of Kazuyo Sejima and the professional trajectory of Sejima-SANAA. And following them there is another one containing transversal texts on form, place and space. The thesis ends with a synthesis on conclusions. The formal strategies are displayed in three chapters. The first one, `Early formal strategies' groups the first phase projects by Sejima. The second one, ‘Formal strategies of Gifu’s paradigm’, is entirely dedicated to the Gifu apartments project, 1994-98, which according to this thesis meant an important inflexion point in Sejima’s trajectory; so much so that the third chapter is named `Formal strategies after Gifu' and gathers the selected projects that followed it. The ‘Early formal strategies', diverse and tentative, move in general around two well-known projectual composition methods ‘composition by parts’, and ‘systematic composition’. This last one –systematic composition- begins and leads in SANAA’s trajectory an aspect which will remain relevant from here on: the understanding of the project as if it were an specific instance of a generic proposal in which -below and beyond the project tangible reality- there lays a logic that could be applicable at other places, for other dimensions, even with other programs; from each project, other projects of the same family could rise. The set of projects using this systematic composition method include, among others, the ‘Platform II House, based on the definition of a constructive element and of rules having to do with its replicas and their possible groupings. It also includes the Saishunkan Seiyaku Women Residence -project that launched Sejima to international fame- that could also be seen as a system, but of a different kind: a system based on the regular repetition of a series of elements along a directive line, thus generating a hypothetical infinite container of which the project would be only a fragment. The formal strategy of the Gifu apartments building would push further towards the generic project concept, adopting the logic of a game. The project would be a bout, a round, one play…, but not the only possible one; others could be played. The thesis confirms this game hypothesis -after having formulated `The Game of Gifu' and identified its elements (board, chips, rules and procedures)- playing the one play from which the building as projected by Sejima would have raised. Gifu extends the concept of ‘repeating a constructive element’ to that of ‘repeating a space pattern element’, and to what it implies: the decoupling of form and function, leading to a new concept of flexibility that no longer refers to the flexible use of the constructed building but to the projectual moment at which the specific functions are assigned to the space patterns. This thesis proposes that this allocation of functions would be one of the last steps in projectual process, quite opposite from the modern premise: “form follows function”. The Formal strategies after Gifu do also have a game logic; but, as their names reveal, each strategy responds to a different game: ‘Game Boards’, present with different maturity levels in several projects; ‘Elements from a Catalogue’, in the Kanazawa Museum; ‘Aprioristic Form’, in the Flower House; and ‘Repetition of a topologic situation', in the Rolex Center. All of these strategies, or games, maintain common aspects having to do with architectural form; aspects that were already present, precisely, in Gifu: repetition of space pattern units, uncoupling of form and function, and a new meaning of flexibility. -`Game Boards’ consists on setting up a base geometry -each project his, generally reticular- and give form to each project system (structure, closings, partitions and furniture) by choosing elements -intersections, lines, modules- it offers. Each project system is formed, in principle, with no subordinated relation with any of the others; when subordination is unavoidable, the game rules determine that the load bearing structural system may not be the one to materialize the base geometric order, which means that it does not exert the dominant role. Therefore, ‘Game Boards' transgresses the Modern logic, because the structure neither reflects nor reveals the base order, and because the systems do not respect any of the hierarchic and chained subordination relations that the ‘free plan’ called for. ‘Game Boards' leads to quite different solutions and formal projects: the Oogaki and Park Coffee projects show incipient Game Boards; The Almere Kunstlinie and the Zollverein School present consolidations of this strategy; and the Toledo’s Glass Pavilion results from subverting the strategy. In addition, the Toledo project takes the repetition concept beyond that of using a constructive element and a space pattern element (in this case with a bubble form) to end up affecting the personal experience of the spectator, who, wherever he is, feels to always be in the same place. This thesis denominates that repetitive space as ‘Mantra space '. -‘Elements from a Catalogue’ is shown with the Kanazawa Museum. Its logic starts from the definition of a series of elements, very few, and it is based on the huge number of possible combinations among them. The ‘Elements from a Catalogue’ approach was announced in the Gifu project when characterizing its space pattern elements. -Aprioristic Form' is illustrated by the Flower House. The decision on the type of form -in this case the form of an amoeba- would be the beginning of the projectual process, but it does not mean it is arbitrary form: the amoeba form implies repeating a space pattern (pseudopodia) and an apotheosis of the repetition concept: embracing the space experience, it gives rise to a repetitive or mantra space. ‘Mantra Space’ is one of leitmotivs used as an argument in the last formal strategy Thesis decodes: the Rolex Center. With respect to the ‘Projectual strategies of the load bearing structure’, the thesis finds and traces a trajectory of five projectual strategies: ‘preeminence, concealment, dissolution, disappearance and desvirtuación’. --Preeminence is present in Sejima’s first works in which she resorts to structures which have a dominant preeminent role in the project in so far as they impersonate the greater scale and/or materialize the base geometric order. In later works that preeminence will be inverted, the projects aiming towards its opposite: lighter, slighter, smaller structures. -Concealment reduces the dominant role of the structure. At the outset concealment is literal, almost hiding the structural elements, as in Gifu; soon it will become more sophisticated, such as the concealment by camouflage or the paradoxical concealment by multiplication in the Koga Park Café. -Dissolution diminishes the dominant condition of the structure: instead of its’ being configured as unitary or homogenous system is fragmented in several subsystems. -Disappearance talks about structures that fade away as self referred and independent systems; projects in which the load bearing function is carried out by other systems such as the set of partitions. Disappearance reaches its zenith at the Flower House, whose perimeter functions structurally being, in addition, transparent, immaterial: its structure has become invisible, has disappeared. -Desvirtuación talks about structures that do appear like independent self-systems, but which that do not longer have a preeminent paper, inasmuch as they do not materialize the base order. This strategy correlates with the ‘Game Boards’ formal strategy. The thesis conclusions are show by the organization of the thesis itself: its identification of the different strategies. Even so, as epilogues, the thesis exposes six ‘Conclusions’. The first two emphasize the leading thread of the work done, rooted in the generic quality of the Sejima-SANAA projectual strategies. The following four expound to what extent their projects show features, or formal and/or structural signifiers, which also are or can be read as characteristic signals of the contemporary architectonic panorama, and raise the key question: aiming farther, may some of them be taken as original contributions? -As original contributions the conclusions highlight: the identification between the generic ideal and the concrete project; and the proposal of a new, hybrid space, kind of an intermediate stage between the traditional subdivided compartmented space and the continuous modern. -As symptoms of contemporaneousness: in relation to the form it highlights the transferring of the formal specificity from the part to the whole; and in relation to the structure, it underscore the contemporary tendency towards lighter and growingly slimmer structures, tending to the evanescent. This last one, the tendency towards structural evanescence, could have condition of being an original contribution, not in vain it carries the structural disappearance towards its last consequences; and in the case of structures with physical presence, it makes them to cease being the ordering system orchestrating the projectual process.

Crecimiento forestal en el bosque tropical de montaña: efectos de la diversidad florística y de la manipulación de nutrientes

El bosque tropical de montaña, es considerado zona de mega diversidad y de alto grado de endemismo, por las diferentes zonas ecológicas que presenta. Durante las últimas décadas estos bosques han recibido mayor atención por parte de investigadores, sin embargo, aún existe poca investigación en determinar cuáles son las respuestas de los bosques a los cambios ambientales a los que son sometidos. Estos bosques están sufriendo serias amenazas como pérdida de cobertura vegetal y cambios en los ciclos de nutrientes. El trabajo se dividió en cuatro objetivos específicos. i) Caracterización y análisis de patrones altitudinales de la riqueza de especies en el bosque tropical de montaña en el sur del Ecuador; con el fin de conocer cómo varía la diversidad de especies riqueza lo largo de un gradiente altitudinal. ii) Conocer los patrones espaciales del crecimiento en tres remanentes boscosos de un bosque tropical de montaña para determinar cómo la vecindad y la semejanza funcional de ésta influyen en el crecimiento forestal. iii) Conocer los efectos de la fertilización en el crecimiento diamétrico de especies arbóreas, en el bosque tropical de montaña; se analizó cómo reaccionan los árboles a la adición de nutrientes N y P en tres tipos de bosque. iv) Saber la respuesta de la comunidad de árboles a la adición de nutrientes en el bosque montano andino; este objetivo se basó con el supuesto de la deficiencia de tres tipos de nutrientes N, P y Ca, en esta formación boscosa y cómo reaccionan los árboles a la adición de nutrientes. El presente trabajo se llevó a cabo, en el bosque tropical de montaña que se encuentra localizada en la parte adyacente del Parque Nacional Podocarpus (PNP) en la cordillera del Consuelo, forma parte de la cadena oriental de los Andes del sur del Ecuador El trabajo de desarrollo entre los años 2008 y 2014. Para abordar el primer objetivo se establecieron 54 parcelas ubicadas aleatoriamente a lo largo de un gradiente altitudinal (3 niveles de altitud) y se e midieron e identificaron todos los individuos mayores a 5cm de DAP. Se construyó una filogenia con Phylocom y se calcularon diferentes componentes de diversidad para cada parcela ( riqueza taxonómica, diversidad filogenética y edad media de las especies). Ajustando modelos lineares se contrastó el efecto de la altitud sobre dichos componentes y se vio que la riqueza taxonómica y la edad media de las especies aumentaron con la altitud, en sentido contrario a las predicciones de la "hipótesis del conservadurismo tropical" (Tropical Conservatism Hypothesis). Para abordar el segundo objetivo se realizó una remedición de todos los árboles cartografiados en tres parcelas permanentes de alrededor de 5000 m2 cada una, representativas de tres estados diferentes de la sucesión del bosque montano. A partir de las coordenadas y de los datos de registrados, y empleando diferentes funciones de correlación de marca se analizó la distribución espacial del tamaño y del crecimiento relativo y del tamaño. Se constató que mientras que el tamaño de los árboles presentó una correlación espacial negativa, el crecimiento presentó correlación espacial positiva, en ambos casos a distancias cortas. El rango y la magnitud de ambas correlaciones aumentaron al avanzar la sucesión. La distribución espacial del crecimiento mostró una correlación negativa con la distribución espacial de tamaños. Por otro lado, la distribución espacial del crecimiento mostró una correlación negativa para árboles semejantes funcionalmente y positiva cuando se calculó entre árboles con diferente estrategia funcional. En conjunto, los resultados obtenidos señalan un aumento de la importancia de procesos competitivos y una mayor estructuración espacial del crecimiento y de la distribución de tamaños al avanzar la sucesión. Para el tercer y cuarto objetivo se instalaron 52 parcelas distribuidas en bloques donde se fertilizaron dos veces al año durante 6,4 años, se identificaron todos los individuos mayores a 10 cm de DAP, y se midió el crecimiento diamétrico durante estos años Con la adición de nutrientes realizada a los diferentes tipos de bosque en la gradiente altitudinal, encontramos que el efecto sobre el crecimiento diamétrico en la comunidad varia con el rango altitudinal, y el tipo de nutriente, analizando a nivel de las especies, en la mayoría de los casos las especies comunes no tuvieron cambios significativos a la adición de nutrientes. Los resultados de este estudio aportan nuevas evidencias para el entendimiento de la diversidad, estructura y dinámica de los bosques tropicales de montaña. ABSTRACT The montane tropical forest is considered a megadiverse habitat that harbor an enormous degree of endemism. This is mainly due to the high degree of environmental heterogeneity found and the presence of different well defined ecological areas. These forests have received more attention during the last decades, however, the information regarding the responses of these forests to environmental change, is still scarce. These forests are seriously endangered and are suffering serious threats, such as loss of vegetative cover, changes in the nutrient cycles. The work was divided in four specific objectives: i) Characterization and analysis of the species richness altitudinal patterns in the montane tropical forest of south Ecuador. Specifically, how species diversity changes along altitudinal gradients. ii) Exploring the spatial patterns of tree growth in three remnants of a montane tropical forest, and analyze how tree neighborhood and functional similarity among trees influence tree growth. Tropical Conservatism Hypothesis iii) Understanding the effects of fertilization in arboreal species growth (increase in diameter) of the montane tropical forest. Specifically we studied the effects of P and N addition on three different forests across an altitudinal gradient. iv) Know the response of the community of trees to the addition of nutrients in the Andean montane forest; this objective was based on the supposition of deficiency of three types of nutrients: P, N and Ca in this forest all formation and how the trees react to the addition of these nutrients. The present work was carried out in the montane tropical forest located in Bombuscaro, San Francisco and Cajanuma close to Podocarpus National Park (PNP) on Consuelo mountain range (Andean oriental range) at South of Ecuador. Field work was carried out during 2008 and 2014. To address the first objective, we randomly placed 54 plots along an altitudinal gradient. In these plots, every individual larger than 5 cm of DBH was measured and identified. A phylogeny was build with Phylocom and different diversity components (taxonomic richness, phylogenetic diversity and average species age) were computed for each plot. Linear models were used to test the effects of altitude on the diversity components. Our results showed that, contrary to the Tropical Conservatism Hypothesis, both taxonomic richness and average species age increased with altitude. To address our second objective, all mapped trees in three successional permanent plots (around ~5000 m2 each) were re-measured. Using different mark correlation functions, we analyzed the spatial distribution of tree-size and tree relative growth rate. Whereas tree size showed negative spatial correlation at fine spatial scales, relative growth rate showed positive correlation at the same scales. The range and magnitude of those correlations increased along successional stage. The spatial distribution of the relative growth rate was negatively correlated with the spatial distribution of tree sizes. Additionally, we found that the spatial correlation of the relative growth rate was negative for functionally similar trees and positive when computed for functionally different trees. In synthesis, our results point to an increase of competitive processes and strong spatial structure of relative growth rate and tree size along succession. For the third and fourth objectives, 52 plots were placed in a block design and were fertilized twice a year for 6,4 years. In these plots all the individuals with DBH > 10 cm were identified, and the diametrical growth was measured during these years. The nutrient addition at the three different altitude forests, revealed that the effect on the diametrical growth in the community varied with the altitudinal range. When analyzed at species level, the addition of nutrients was no significant in most cases. These results represent new evidences that will improved our understanding of diversity patterns and structure, and the dynamics of tropical montane forests.

BAC-VAC, a novel generation of (DNA) vaccines: A bacterial artificial chromosome (BAC) containing a replication-competent, packaging-defective virus genome induces protective immunity against herpes simplex virus 1

This study aimed to exploit bacterial artificial chromosomes (BAC) as large antigen-capacity DNA vaccines (BAC-VAC) against complex pathogens, such as herpes simplex virus 1 (HSV-1). The 152-kbp HSV-1 genome recently has been cloned as an F-plasmid-based BAC in Escherichia coli (fHSV), which can efficiently produce infectious virus progeny upon transfection into mammalian cells. A safe modification of fHSV, fHSVΔpac, does not give rise to progeny virus because the signals necessary to package DNA into virions have been excluded. However, in mammalian cells fHSVΔpac DNA can still replicate, express the HSV-1 genes, cause cytotoxic effects, and produce virus-like particles. Because these functions mimic the lytic cycle of the HSV-1 infection, fHSVΔpac was expected to stimulate the immune system as efficiently as a modified live virus vaccine. To test this hypothesis, mice were immunized with fHSVΔpac DNA applied intradermally by gold-particle bombardment, and the immune responses were compared with those induced by infection with disabled infectious single cycle HSV-1. Immunization with either fHSVΔpac or disabled infectious single cycle HSV-1 induced the priming of HSV-1-specific cytotoxic T cells and the production of virus-specific antibodies and conferred protection against intracerebral injection of wild-type HSV-1 at a dose of 200 LD50. Protection probably was cell-mediated, as transfer of serum from immunized mice did not protect naive animals. We conclude that BAC-VACs per se, or in combination with genetic elements that support replicative amplification of the DNA in the cell nucleus, represent a useful new generation of DNA-based vaccination strategies for many viral and nonviral antigens.

β1-Integrin Cytoplasmic Subdomains Involved in Dominant Negative Function

The β1-integrin cytoplasmic domain consists of a membrane proximal subdomain common to the four known isoforms (“common” region) and a distal subdomain specific for each isoform (“variable” region). To investigate in detail the role of these subdomains in integrin-dependent cellular functions, we used β1A and β1B isoforms as well as four mutants lacking the entire cytoplasmic domain (β1TR), the variable region (β1COM), or the common region (β1ΔCOM-B and β1ΔCOM-A). By expressing these constructs in Chinese hamster ovary and β1 integrin-deficient GD25 cells (Wennerberg et al., J Cell Biol 132, 227–238, 1996), we show that β1B, β1COM, β1ΔCOM-B, and β1ΔCOM-A molecules are unable to support efficient cell adhesion to matrix proteins. On exposure to Mn++ ions, however, β1B, but none of the mutants, can mediate cell adhesion, indicating specific functional properties of this isoform. Analysis of adhesive functions of transfected cells shows that β1B interferes in a dominant negative manner with β1A and β3/β5 integrins in cell spreading, focal adhesion formation, focal adhesion kinase tyrosine phosphorylation, and fibronectin matrix assembly. None of the β1 mutants tested shows this property, indicating that the dominant negative effect depends on the specific combination of common and B subdomains, rather than from the absence of the A subdomain in the β1B isoform.

Association of Smads with lymphoid enhancer binding factor 1/T cell-specific factor mediates cooperative signaling by the transforming growth factor-β and Wnt pathways

The transforming growth factor-β (TGFβ) and Wnt/wingless pathways play pivotal roles in tissue specification during development. Activation of Smads, the effectors of TGFβ superfamily signals, results in Smad translocation from the cytoplasm into the nucleus where they act as transcriptional comodulators to regulate target gene expression. Wnt/wingless signals are mediated by the DNA-binding HMG box transcription factors lymphoid enhancer binding factor 1/T cell-specific factor (LEF1/TCF) and their coactivator β-catenin. Herein, we show that Smad3 physically interacts with the HMG box domain of LEF1 and that TGFβ and Wnt pathways synergize to activate transcription of the Xenopus homeobox gene twin (Xtwn). Disruption of specific Smad and LEF1/TCF DNA-binding sites in the promoter abrogates synergistic activation of the promoter. Consistent with this observation, introduction of Smad sites into a TGFβ-insensitive LEF1/TCF target gene confers cooperative TGFβ and Wnt responsiveness to the promoter. Furthermore, we demonstrate that TGFβ-dependent activation of LEF1/TCF target genes can occur in the absence of β-catenin binding to LEF1/TCF and requires both Smad and LEF1/TCF DNA-binding sites in the Xtwn promoter. Thus, our results show that TGFβ and Wnt signaling pathways can independently or cooperatively regulate LEF1/TCF target genes and suggest a model for how these pathways can synergistically activate target genes.

A mutant allele of essential, general translation initiation factor DED1 selectively inhibits translation of a viral mRNA

Positive-strand RNA virus genomes are substrates for translation, RNA replication, and encapsidation. To identify host factors involved in these functions, we used the ability of brome mosaic virus (BMV) RNA to replicate in yeast. We report herein identification of a mutation in the essential yeast gene DED1 that inhibited BMV RNA replication but not yeast growth. DED1 encodes a DEAD (Asp-Glu-Ala-Asp)-box RNA helicase required for translation initiation of all yeast mRNAs. Inhibition of BMV RNA replication by the mutant DED1 allele (ded1–18) resulted from inhibited expression of viral polymerase-like protein 2a, encoded by BMV RNA2. Inhibition of RNA2 translation was selective, with no effect on general cellular translation or translation of BMV RNA1-encoded replication factor 1a, and was independent of p20, a cellular antagonist of DED1 function in translation. Inhibition of RNA2 translation in ded1–18 yeast required the RNA2 5′ noncoding region (NCR), which also conferred a ded1–18-specific reduction in expression on a reporter gene mRNA. Comparison of the similar RNA1 and RNA2 5′ NCRs identified a 31-nucleotide RNA2-specific region that was required for the ded1–18-specific RNA2 translation block and attenuated RNA2 translation in wild-type yeast. Further comparisons and RNA structure predictions suggest a modular arrangement of replication and translation signals in RNA1 and RNA2 5′ NCRs that appears conserved among bromoviruses. The 5′ attenuator and DED1 dependence of RNA2 suggest that, despite its divided genome, BMV regulates polymerase translation relative to other replication factors, just as many single-component RNA viruses use translational read-through and frameshift mechanisms to down-regulate polymerase. The results show that a DEAD-box helicase can selectively activate translation of a specific mRNA and may provide a paradigm for translational regulation by other members of the ubiquitous DEAD-box RNA helicase family.

Selectively enhanced contextual fear conditioning in mice lacking the transcriptional regulator CCAAT/enhancer binding protein δ

CCAAT/enhancer binding protein δ (C/EBPδ) is a transcriptional regulator implicated in the hepatic acute phase response and in adipogenic and myeloid cell differentiation. We found that C/EBPδ is widely expressed in the peripheral and central nervous systems, including neurons of the hippocampal formation, indicating a role in neural functions. To examine the role of C/EBPδ in vivo, we generated mice with a targeted deletion of the C/EBPδ gene. This mutation does not interfere with normal embryonic and postnatal development. Performance in a battery of behavioral tests indicates that basic neurological functions are normal. Furthermore, performance in a Morris water maze task suggests that C/EBPδ mutant mice have normal spatial learning. However, in the contextual and auditory-cue-conditioned fear task, C/EBPδ null mice displayed significantly more conditioned freezing to the test context than did wild-type controls, but equivalent conditioning to the auditory cue. These data demonstrate a selectively enhanced contextual fear response in mice carrying a targeted genomic mutation and implicate C/EBPδ in the regulation of a specific type of learning and memory.

Occupational class and cause specific mortality in middle aged men in 11 European countries: comparison of population based studies

Objectives: To compare countries in western Europe with respect to class differences in mortality from specific causes of death and to assess the contributions these causes make to class differences in total mortality.

A Cyanobacterium Lacking Iron Superoxide Dismutase Is Sensitized to Oxidative Stress Induced with Methyl Viologen but Is Not Sensitized to Oxidative Stress Induced with Norflurazon1

A strain of Synechococcus sp. strain PCC 7942 with no functional Fe superoxide dismutase (SOD), designated sodB−, was characterized by its growth rate, photosynthetic pigments, and cyclic photosynthetic electron transport activity when treated with methyl viologen or norflurazon (NF). In their unstressed conditions, both the sodB− and wild-type strains had similar chlorophyll and carotenoid contents and catalase activity, but the wild type had a faster growth rate and higher cyclic electron transport activity. The sodB− was very sensitive to methyl viologen, indicating a specific role for the FeSOD in protection against superoxide generated in the cytosol. In contrast, the sodB− mutant was less sensitive than the wild type to oxidative stress imposed with NF. This suggests that the FeSOD does not protect the cell from excited singlet-state oxygen generated within the thylakoid membrane. Another up-regulated antioxidant, possibly the MnSOD, may confer protection against NF in the sodB− strain. These results support the hypothesis that different SODs have specific protective functions within the cell.

Essential role for complex N-glycans in forming an organized layer of bronchial epithelium.

Mice lacking the complex subset of N-glycans due to inactivation of the Mgat1 gene die at mid-gestation, making it difficult to identify specific biological functions for this class of cell surface carbohydrates. To circumvent this embryonic lethality and to uncover tissue-specific functions for complex N-glycans, WW6 embryonic stem cells with inactivated Mgat1 alleles were tracked in chimeric embryos. The Mgat1 gene encodes N-acetylglucosaminyltransferase I (Glc-NAc-TI; EC 2.4.1.101), the transferase that initiates the synthesis of complex N-glycans. WW6 cells carry an inert beta-globin transgene that allows their identification in chimeras by DNA-DNA in situ hybridization. Independent Mgat1-/- and Mgat1+/- mutant WW6 isolates contributed like parent WW6 cells to the tissues of embryonic day (E) 10.5 to E16.5 chimeras. However, a cell type-specific difference was observed in lung. Homozygous null Mgat1-/- WW6 cells did not contribute to the epithelial layer in more than 99% bronchi. This deficiency was corrected by transfection of a Mgat1 transgene. Interestingly, heterozygous Mgat1+/- WW6 cells were also deficient in populating the layer of bronchial epithelium. Furthermore, examination of lung bud in E9.5 Mgat1-/- mutant embryos showed complete absence of an organized epithelial cell layer in the bronchus. Thus, complex N-glycans are required to form a morphologically recognizable bronchial epithelium, revealing an in vivo, cell type-specific function for this class of N-glycans.

Nucleotide-specific interaction of Ran/TC4 with nuclear transport factors NTF2 and p97.

The use of permeabilized cell models to study nuclear protein import has led to the identification of cytosolic components of the import machinery, including the NLS receptor, p97, Ran/TC4, and nuclear transport factor 2 (NTF2). These proteins are required to reconstitute docking of transport ligand at the nuclear pore complex and subsequent translocation through the nuclear pore. However, a detailed molecular understanding of how these factors mediate protein import is lacking. Here we describe the results of solution and solid phase binding assays, which demonstrate that the small GTPase Ran/TC4 interacts directly with the cytosolic transport factors p97 and NTF2. By preloading recombinant Ran/TC4 with [gamma-32P]GTP or [3H]GDP, we show that the interactions with p97 and NTF2 are specific for the GTP- and GDP-bound forms, respectively. These data together with previous studies lead us to suggest that the interaction of the GTP-bound form of Ran/TC4 with p97 is linked to an early step in the nuclear protein import pathway and that the association of the GDP-bound form of Ran/TC4 with NTF2 helps define vectorial transport.

Preferential induction of a Th1 immune response and inhibition of specific IgE antibody formation by plasmid DNA immunization.

We compared the antigen-specific antibody isotypes and lymphokine secretion by CD4+ T cells in BALB/c mice immunized intradermally with either Escherichia coli beta-galactosidase (beta-gal) or plasmid DNA (pDNA) encoding beta-gal in a cytomegalovirus-based expression vector (pCMV-LacZ). pCMV-LacZ induced mainly IgG2a, whereas beta-gal in saline or alum induced IgG1 and IgE beta-gal-specific antibodies. In addition, splenic CD4+ T helper (Th) cells isolated from pDNA-immunized mice secreted interferon-gamma but not interleukin (IL)-4 and IL-5, whereas Th cells from beta-gal-injected mice secreted IL-4 and IL-5 but not interferon-gamma after in vitro stimulation with antigen. Together these data demonstrate that pDNA immunization induced a T helper type 1 (Th1) response, whereas protein immunization induced a T helper type 2 (Th2) response to the same antigen. Interestingly, priming of mice with pCMV-LacZ prevented IgE antibody formation to a subsequent i.p. beta-gal in alum injection. This effect was antigen-specific, because priming with pCMV-LacZ did not inhibit IgE anti-ovalbumin antibody formation. Most importantly, intradermal immunization with pCMV-LacZ (but not pCMV-OVA) of beta-gal in alum-primed mice caused a 66-75% reduction of the IgE anti-beta-gal titer in 6 weeks. Also, pCMV-LacZ induced specific IgG2a antibody titers and interferon-gamma secretion by Th cells in the beta-gal in alum-primed mice. The data demonstrate that gene immunization induces a Th1 response that dominates over an ongoing protein-induced Th2 response in an antigen-specific manner. This suggests that immunization with pDNA encoding for allergens may provide a novel type of immunotherapy for allergic diseases.

Visualization of the vesicular acetylcholine transporter in cholinergic nerve terminals and its targeting to a specific population of small synaptic vesicles.

Immunohistochemical visualization of the rat vesicular acetylcholine transporter (VAChT) in cholinergic neurons and nerve terminals has been compared to that for choline acetyltransferase (ChAT), heretofore the most specific marker for cholinergic neurons. VAChT-positive cell bodies were visualized in cerebral cortex, basal forebrain, medial habenula, striatum, brain stem, and spinal cord by using a polyclonal anti-VAChT antiserum. VAChT-immuno-reactive fibers and terminals were also visualized in these regions and in hippocampus, at neuromuscular junctions within skeletal muscle, and in sympathetic and parasympathetic autonomic ganglia and target tissues. Cholinergic nerve terminals contain more VAChT than ChAT immunoreactivity after routine fixation, consistent with a concentration of VAChT within terminal neuronal arborizations in which secretory vesicles are clustered. These include VAChT-positive terminals of the median eminence or the hypothalamus, not observed with ChAT antiserum after routine fixation. Subcellular localization of VAChT in specific organelles in neuronal cells was examined by immunoelectron microscopy in a rat neuronal cell line (PC 12-c4) expressing VAChT as well as the endocrine and neuronal forms of the vesicular monoamine transporters (VMAT1 and VMAT2). VAChT is targeted to small synaptic vesicles, while VMAT1 is found mainly but not exclusively on large dense-core vesicles. VMAT2 is found on large dense-core vesicles but not on the small synaptic vesicles that contain VAChT in PC12-c4 cells, despite the presence of VMAT2 immunoreactivity in central and peripheral nerve terminals known to contain monoamines in small synaptic vesicles. Thus, VAChT and VMAT2 may be specific markers for "cholinergic" and "adrenergic" small synaptic vesicles, with the latter not expressed in nonstimulated neuronally differentiated PC12-c4 cells.

Tissue-specific response of the human platelet-activating factor receptor gene to retinoic acid and thyroid hormone by alternative promoter usage.

We have studied the effects of retinoic acid (RA) and thyroid hormone (3,3',5-triiodothyronine; T3) on platelet-activating factor receptor (PAFR) gene expression in intact rats and the ability of two human PAFR gene promoters (PAFR promoters 1 and 2) to generate two transcripts (PAFR transcripts 1 and 2). Northern blotting showed that RA and T3 regulated PAFR gene expression only in rat tissues that express PAFR transcript 2. Functional analysis of the human PAFR promoter 2 revealed that responsiveness to RA and T3 was conferred through a 24-bp element [PAFR-hormone response element (HRE) located from -67 to -44 bp of the transcription start site, whereas PAFR promoter 1 did not respond to these hormones. The PAFR-HRE is composed of three direct repeated TGACCT-like hexamer motifs with 2-and 4-bp spaces, and the two upstream and two downstream motifs were identified as response elements for RA and T3. Thus, the PAF-PAFR pathway is regulated by the PAFR level altered by a tissue-specific response to RA and T3 through the PAFR-HRE of the PAFR promoter 2.

Gene disruptions using P transposable elements: an integral component of the Drosophila genome project.

Biologists require genetic as well as molecular tools to decipher genomic information and ultimately to understand gene function. The Berkeley Drosophila Genome Project is addressing these needs with a massive gene disruption project that uses individual, genetically engineered P transposable elements to target open reading frames throughout the Drosophila genome. DNA flanking the insertions is sequenced, thereby placing an extensive series of genetic markers on the physical genomic map and associating insertions with specific open reading frames and genes. Insertions from the collection now lie within or near most Drosophila genes, greatly reducing the time required to identify new mutations and analyze gene functions. Information revealed from these studies about P element site specificity is being used to target the remaining open reading frames.