Proyectando entre ruinas : lógicas de intervención continua en la arquitectura contemporánea

Los cambios percibidos hacia finales del siglo XX y a principios del nuevo milenio, nos ha mostrado que la crisis cultural de la que somos participes refleja también una crisis de los modelos universales. Nuestra situación contemporánea, parece indicar que ya no es posible formular un sistema estético para atribuirle una vigencia universal e intemporal más allá de su estricta eficacia puntual. La referencia organizada, delimitada, invariable y específica que ofrecía cualquier emplazamiento, en tanto preexistencia, reflejaba una jerarquía del sistema formal basado en lo extensivo: la medida, las normas, el movimiento, el tiempo, la modulación, los códigos y las reglas. Sin embargo, actualmente, algunos aspectos que permanecían latentes sobre lo construido, emergen bajo connotaciones intensivas, transgrediendo la simple manifestación visual y expresiva, para centrase en las propiedades del comportamiento de la materia y la energía como determinantes de un proceso de adaptación en el entorno. A lo largo del todo el siglo XX, el desarrollo de la relación del proyecto sobre lo construido ha sido abordado, casi en exclusiva, entre acciones de preservación o intervención. Ambas perspectivas, manifestaban esfuerzos por articular un pensamiento que diera una consistencia teórica, como soporte para la producción de la acción aditiva. No obstante, en las últimas décadas de finales de siglo, la teoría arquitectónica terminó por incluir pensamientos de otros campos que parecen contaminar la visión sesgada que nos refería lo construido. Todo este entramado conceptual previo, aglomeraba valiosos intentos por dar contenido a una teoría que pudiese ser entendida desde una sola posición argumental. Es así, que en 1979 Ignasi Solá-Morales integró todas las imprecisiones que referían una actuación sobre una arquitectura existente, bajo el termino de “intervención”, el cual fue argumentado en dos sentidos: El primero referido a cualquier tipo de actuación que se puede hacer en un edificio, desde la defensa, preservación, conservación, reutilización, y demás acciones. Se trata de un ámbito donde permanece latente el sentido de intensidad, como factor común de entendimiento de una misma acción. En segundo lugar, más restringido, la idea de intervención se erige como el acto crítico a las ideas anteriores. Ambos representan en definitiva, formas de interpretación de un nuevo discurso. “Una intervención, es tanto como intentar que el edificio vuelva a decir algo o lo diga en una determinada dirección”. A mediados de 1985, motivado por la corriente de revisión historiográfica y la preocupación del deterioro de los centros históricos que recorría toda Europa, Solá-Morales se propone reflexionar sobre “la relación” entre una intervención de nueva arquitectura y la arquitectura previamente existente. Relación condicionada estrictamente bajo consideraciones lingüísticas, a su entender, en sintonía con toda la producción arquitectónica de todo el siglo XX. Del Contraste a la Analogía, resumirá las transformaciones en la concepción discursiva de la intervención arquitectónica, como un fenómeno cambiante en función de los valores culturales, pero a su vez, mostrando una clara tendencia dialógica entres dos categorías formales: El Contraste, enfatizando las posibilidades de la novedad y la diferencia; y por otro lado la emergente Analogía, como una nueva sensibilidad de interpretación del edificio antiguo, donde la semejanza y la diversidad se manifiestan simultáneamente. El aporte reflexivo de los escritos de Solá-Morales podría ser definitivo, si en las últimas décadas antes del fin de siglo, no se hubiesen percibido ciertos cambios sobre la continuidad de la expresión lingüística que fomentaba la arquitectura, hacia una especie de hipertrofia figurativa. Entre muchos argumentos: La disolución de la consistencia compositiva y el estilo unitario, la incorporación volumétrica del proyecto como dispositivo reactivo, y el cambio de visión desde lo retrospectivo hacia lo prospectivo que sugiere la nueva conservación. En este contexto de desintegración, el proyecto, en tanto incorporación o añadido sobre un edificio construido, deja de ser considerado como un apéndice volumétrico subordinado por la reglas compositivas y formales de lo antiguo, para ser considerado como un organismo de orden reactivo, que produce en el soporte existente una alteración en su conformación estructural y sistémica. La extensión, antes espacial, se considera ahora una extensión sensorial y morfológica con la implementación de la tecnología y la hiper-información, pero a su vez, marcados por una fuerte tendencia de optimización energética en su rol operativo, ante el surgimiento del factor ecológico en la producción contemporánea. En una sociedad, como la nuestra, que se está modernizando intensamente, es difícil compartir una adecuada sintonía con las formas del pasado. Desde 1790, fecha de la primera convención francesa para la conservación de monumentos, la escala de lo que se pretende preservar es cada vez más ambiciosa, tanto es así, que al día de hoy el repertorio de lo que se conserva incluye prácticamente todas las tipologías del entorno construido. Para Koolhaas, el intervalo entre el objeto y el momento en el cual se decide su conservación se ha reducido, desde dos milenios en 1882 a unas décadas hoy en día. En breve este lapso desaparecerá, demostrando un cambio radical desde lo retrospectivo hacia lo prospectivo, es decir, que dentro de poco habrá que decidir que es lo que se conserva antes de construir. Solá-Morales, en su momento, distinguió la relación entre lo nuevo y lo antiguo, entre el contraste y la analogía. Hoy casi tres décadas después, el objetivo consiste en evaluar si el modelo de intervención arquitectónica sobre lo construido se ha mantenido desde entonces o si han aparecido nuevas formas de posicionamiento del proyecto sobre lo construido. Nuestro trabajo pretende demostrar el cambio de enfoque proyectual con la preexistencia y que éste tiene estrecha relación con la incorporación de nuevos conceptos, técnicas, herramientas y necesidades que imprimen el contexto cultural, producido por el cambio de siglo. Esta suposición nos orienta a establecer un paralelismo arquitectónico entre los modos de relación en que se manifiesta lo nuevo, entre una posición comúnmente asumida (Tópica), genérica y ortodoxa, fundamentada en lo visual y expresivo de las últimas décadas del siglo XX, y una realidad emergente (Heterotópica), extraordinaria y heterodoxa que estimula lo inmaterial y que parece emerger con creciente intensidad en el siglo XXI. Si a lo largo de todo el siglo XX, el proyecto de intervención arquitectónico, se debatía entre la continuidad y discontinuidad de las categorías formales marcadas por la expresión del edificio preexistente, la nueva intervención contemporánea, como dispositivo reactivo en el paisaje y en el territorio, demanda una absoluta continuidad, ya no visual, expresiva, ni funcional, sino una continuidad fisiológica de adaptación y cambio con la propia dinámica del territorio, bajo nuevas reglas de juego y desplegando planes y estrategias operativas (proyectivas) desde su propia lógica y contingencia. El objeto de esta investigación es determinar los nuevos modos de continuidad y las posibles lógicas de producción que se manifiestan dentro de la Intervención Arquitectónica, intentando superar lo aparente de su relación física y visual, como resultado de la incorporación del factor operativo desplegado por el nuevo dispositivo contemporáneo. Creemos que es acertado mantener la senda connotativa que marca la denominación intervención arquitectónica, por aglutinar conceptos y acercamientos teóricos previos que han ido evolucionando en el tiempo. Si bien el término adolece de mayor alcance operativo desde su formulación, una cualidad que infieren nuestras lógicas contemporáneas, podría ser la reformulación y consolidación de un concepto de intervención más idóneo con nuestros tiempos, anteponiendo un procedimiento lógico desde su propia necesidad y contingencia. Finalmente, nuestro planteamiento inicial aspira a constituir un nueva forma de reflexión que nos permita comprender las complejas implicaciones que infiere la nueva arquitectura sobre la preexistencia, motivada por las incorporación de factores externos al simple juicio formal y expresivo preponderante a finales del siglo XX. Del mismo modo, nuestro camino propuesto, como alternativa, permite proyectar posibles sendas de prospección, al considerar lo preexistente como un ámbito que abarca la totalidad del territorio con dinámicas emergentes de cambio, y con ellas, sus lógicas de intervención.Abstract The perceived changes towards the end of the XXth century and at the beginning of the new milennium have shown us that the cultural crisis in which we participate also reflects a crisis of the universal models. The difference between our contemporary situation and the typical situations of modern orthodoxy and post-modernistic fragmentation, seems to indicate that it is no longer possible to formulate a valid esthetic system, to assign a universal and eternal validity to it beyond its strictly punctual effectiveness; which is even subject to questioning because of the continuous transformations that take place in time and in the sensibility of the subject itself every time it takes over the place. The organised reference that any location offered, limited, invariable and specific, while pre-existing, reflected a hierarchy of the formal system based on the applicable: measure, standards, movement, time, modulation, codes and rules. Authors like Marshall Mc Luhan, Paul Virilio, or Marc Augé anticipated a reality where the conventional system already did not seem to respond to the new architectural requests in which information, speed, disappearance and the virtual had blurred the traditional limits of place; pre-existence did no longer possess a specific delimitation and, on the contrary, they expect to reach a global scale. Currently, some aspects that stayed latent relating to the constructed, surface from intensive connotations, transgressing the simple visual and expressive manifestation in order to focus on the traits of the behaviour of material and energy as determinants of a process of adaptation to the surroundings. Throughout the entire Century, the development of the relation of the project relating to the constructed has been addressed, almost exclusively, in preservational or interventianal actions. Both perspectives showed efforts in order to express a thought that would give a theoretical consistency as a base for the production of the additive action. Nevertheless, the last decades of the Century, architectural theory ended up including thoughts from other fields that seem to contaminate the biased vision 15 which the constructed related us. Ecology, planning, philosophy, global economy, etc, suggest new approaches to the construction of the contemporary city; but this time with a determined idea of change and continuous transformation, that enriches the panorama of thought and architectural practice, at the same time, according to some, it puts disciplinary specification at risk, given that there is no architecture without destruction, the constructed organism requires mutation in order to adjust to the change of shape. All of this previous conceptual framework gathered valuable intents to give importance to a theory that could be understood solely from an argumental position. Thusly, in 1979 Ignasi Solá-Morales integrated all of the imprecisions that referred to an action in existing architecture under the term of “Intervention”, which was explained in two ways: The first referring to any type of intervention that can be carried out in a building, regarding protection, conservation, reuse, etc. It is about a scope where the meaning of intensity stays latent as a common factor of the understanding of a single action. Secondly, more limitedly, the idea of intervention is established as the critical act to the other previous ideas such as restauration, conservation, reuse, etc. Both ultimately represent ways of interpretation of a new speech. “An intervention, is as much as trying to make the building say something again or that it be said in a certain direction”. Mid 1985, motivated by the current of historiographical revision and the concerns regarding the deterioration of historical centres that traversed Europe, Solá-Morales decides to reflect on “the relationship” between an intervention of the new architecture and the previously existing architecture. A relationship determined strictly by linguistic considerations, to his understanding, in harmony with all of the architectural production of the XXth century. From Contrast to Analogy would summarise transformations in the discursive perception of architectural intervention, as a changing phenomenon depending on cultural values, but at the same time, showing a clear dialogical tendency between two formal categories: Contrast, emphasising the possibilities of novelty and difference; and on the other hand the emerging Analogy, as a new awareness of interpretation of the ancient building, where the similarity and diversity are manifested simultaneously. For Solá-Morales the analogical procedure is not based on the visible simultaneity of formal orders, but on associations that the subject establishes throughout time. Through analogy it is tried to overcome the simple visual relationship with the antique, to focus on its spacial, physical and geographical nature. If the analogical attempt guides an opening towards a new continuity; it still persists in the connection of dimensional, typological and figurative factors, subordinate to the formal hierarchy of the preexisting subjects. 16 The reflexive contribution of Solá-Morales’ works could be final, if in the last decades before the end of the century there had not been certain changes regarding linguistic expression, encouraged by architecture, towards a kind of figurative hypertrophy, amongst many arguments we are in this case interested in three moments: The dissolution of the compositional consistency and the united style, the volumetric incorporation of the project as a reactive mechanism, and the change of the vision from retrospective towards prospective that the new conservation suggests. The recurrence to the history of architecture and its recognisable forms, as a way of perpetuating memory and establishing a reference, dissolved any instinct of compositive unity and style, provoking permanent relationships to tend to disappear. The composition and coherence lead to suppose a type of discontinuity of isolated objects in which only possible relationships could appear; no longer as an order of certain formal and compositive rules, but as a special way of setting elements in a specific work. The new globalised field required new forms of consistency between the project and the pre-existent subject, motivated amongst others by the higher pace of market evolution, increase of consumer tax and the level of information and competence between different locations; aspects which finally made stylistic consistence inefficient. In this context of disintegration, the project, in incorporation as well as added to a constructed building, stops being considered as a volumetric appendix subordinate to compositive and formal rules of old, to be considered as an organism of reactive order, that causes a change in the structural and systematic configuration of the existing foundation. The extension, previsouly spatial, is now considered a sensorial and morphological extension, with the implementation of technology and hyper-information, but at the same time, marked by a strong tendency of energetic optimization in its operational role, facing the emergence of the ecological factor in contemporary production. The technological world turns into a new nature, a nature that should be analysed from ecological terms; in other words, as an event of transition in the continuous redistribution of energy. In this area, effectiveness is not only determined by the capacity of adaptation to changing conditions, but also by its transforming capacity “expressly” in order to change an environment. In a society, like ours, that is modernising intensively, it is difficult to share an adecuate agreement with the forms of the past. From 1790, the date of the first French convention for the conservation of monuments, the scale of what is expexted to be preserved is more and more ambitious, so much so that nowadays the repertoire of that what is conserved includes practically all typologies of the constructed surroundings. For Koolhaas, the ínterval between the object and the moment when its conservation is decided has been reduced, from two 17 milennia in 1882 to a few decades nowadays. Shortly this lapse will disappear, showing a radical change of retrospective towards prospective, that is to say, that soon it will be necessary to decide what to conserve before constructing. The shapes of cities are the result of the continuous incorporation of architecture, and perhaps that only through architecture the response to the universe can be understood, the continuity of what has already been constructed. Our work is understood also within that system, modifying the field of action and leaving the road ready for the next movement of those that will follow after us. Continuity does not mean conservatism, continuity means being conscient of the transitory value of our answers to specific needs, accepting the change that we have received. That what has been constructed to remain and last, should cause future interventions to be integrated in it. It is necessary to accept continuity as a rule. Solá-Morales, in his time, distinguished between the relationship with new and old, between contrast and analogy. Today, almost three decades later, the objective consists of evaluating whether the model of architectural intervention in the constructed has been maintained since then or if new ways of positioning the project regarding the constructed have appeared. Our work claims to show the change of the approach of projects with pre-existing subjects and that this has got a close relation to the incorporation of new concepts, techniques, tools and necessities that impress the cultural context, caused by the change of centuries. This assumption guides us to establish a parallelism between the forms of connection where that what is new is manifested between a commonly assumed (topical), generic and orthodox position, based on that what is visual and expressive in the last decades of the XXth century, and an emerging (heterotopical), extraordinary and heterodox reality that stimulates the immaterial and that seems to emerge with growing intensity in the XXIst century. If throughout the XXth century the project of architectural intervention was considered from the continuity and discontinuity of formal categories, marked by the expression of the pre-existing building, the new contemporary intervention, as a reactive device in the landscape and territory, demands an absolute continuity. No longer a visual, expressive or functional one but a morphological continuity of adaptation and change with its own territorial dynamics, under new game rules and unfolding new operative (projective) strategies from its own logic and contingency. 18 The aim of this research is to determine new forms of continuity and the possible logic of production that are expressed in the Architectural Intervention, trying to overcome the obviousness of its physical and visual relationship, at the beginning of this new century, as a result of the incorporation of the operative factor that the new architectural device unfolds. We think it is correct to maintain the connotative path that marks the name architectural intervention by bringing previous concepts and theorical approaches that have been evolving through time together. If the name suffers from a wider operational range because of its formulation, a quality that our contemporary logic provokes, the reformulation and consolidation of an interventional concept could be more suitable for our times, giving preference to a logical method from its own necessity and contingency. It seems that now time shapes the topics, it is no longer about materialising a certain time but about expressing the changes that its new temporality generates. Finally, our initial approach aspires to form a new way of reflection that permits us to understand the complex implications that the new architecture submits the pre-existing subject to, motivated by the incorporation of factors external to simple formal and expressive judgement, prevailing at the end of the XXth century. In the same way, our set road, as an alternative, permits the contemplation of possible research paths, considering that what is pre-existing as an area that spans the whole territory with emerging changing dynamics and, with them, their interventional logics.

A fuzzy extension of MAGERIT methodology for risk analysis in information systems

We propose a fuzzy approach to deal with risk analysis for information systems. We extend MAGERIT methodology that valuates the asset dependencies to a fuzzy framework adding fuzzy linguistic terms to valuate the different elements (terminal asset values, asset dependencies as well as the probability of threats and the resulting asset degradation) in risk analysis. Computations are based on the trapezoidal fuzzy numbers associated with these linguistic terms and, finally, the results of these operations are translated into a linguistic term by means of a similarity function.

Comportamiento hidrológico de cuencas de media montaña españolas: efectos de los procesos de acumulación / fusión de nieve en terrenos permeables sobre la infiltración y régimen de caudales Alto Tajo

La infiltración de agua en el suelo y la recarga profunda del agua subterránea contenida en los acuíferos es un proceso lento en relación con otros fenómenos hidrológicos. La redacción de esta tesis ha pretendido contribuir al estudio de la influencia que el almacenamiento de la precipitación sólida en forma de manto de nieve y su eventual fusión puedan tener sobre dicho proceso en áreas de media montaña (1.000 – 2.000 m.) en las que con gran frecuencia se sitúan las cabeceras de los ríos peninsulares. Para ello se ha partido del análisis de las diferentes variables intervinientes durante un determinado periodo temporal y sobre un espacio geográfico concreto, por lo que su metodología es de naturaleza empírica. La extensión del periodo (2002/03 a 2010/11) ha venido condicionada por la disponibilidad de los valores de algunas de sus principales variables, como han sido el equivalente en agua de la nieve acumulada y los caudales procedentes de su fusión. Éstos se han obtenido como resultado de la aplicación del modelo ASTER, desarrollado en el programa de Evaluación de los Recursos Hídricos procedentes de la Innivación (ERHIN), calibrado – entre otros- con datos de precipitaciones, temperatura y caudales provenientes a su vez del Sistema Automático de Información Hidrológica (SAIH). Ambos programas fueron implantados por la Administración en las diferentes Confederaciones Hidrográficas y en determinados Organismos de cuenca actuales, en cuyo desarrollo participó el autor de esta tesis. En cuanto a la zona de estudio se ha procedido a su elección considerando las posibles áreas de media montaña en las que la presencia de la nieve fuera hidrológicamente significativa y estuvieran constituidas litológicamente por afloramientos permeables que no impidieran la infiltración en el terreno y la formación de acuíferos de cierta relevancia. El interés se centró discrecionalmente en la cuenca del Tajo, tanto por el carácter estratégico de la misma -como suministradora en la actualidad de excedentes a otras cuencas deficitarias- como por el valor representativo de sus condiciones climáticas y orográficas en relación con otras cuencas hidrográficas peninsulares. Para ello se partió de las cabeceras de ríos identificadas por el programa ERHIN por su interés nivológico para la implantación del modelo ASTER y de las Masas de Agua Subterráneas MASb (antes Unidades Hidrogeológicas UUHH) definidas en los planes hidrológicos. La intersección en el territorio de ambos criterios condujo, finalmente, a la zona del Alto Tajo, en la que se cumplen ambos requisitos. El tramo quedó concretado en el comprendido entre las cabeceras de los ríos Tajo y Guadiela y la cola de los embalses de Entrepeñas y Buendía respectivamente, puntos de cierre para la calibración llevada a cabo en la modelización ASTER. Gran parte de éste discurre, en su parte alta, sobre rocas carbonatadas (calizas y dolomías del Jurásico y Cretácico), relacionados con las MASb de Tajuña-Montes Universales, Molina de Aragón y Sigüenza-Maranchón. Los valores diarios de las reservas de agua en forma de nieve, evapotranspiración y caudales procedentes de la fusión se han obtenido a partir de los resultados del mencionado modelo, procediéndose al cálculo de la infiltración por balance hídrico durante el periodo de estudio considerado, teniendo en cuenta los valores de precipitación, evapotranspiración y aportaciones de caudales. Esto ha requerido el estudio previo de las condiciones hidrogeológicas de la zona seleccionada con objeto de conocer las posibles interconexiones subterráneas que pudieran alterar los saldos entre las variables intervinientes anteriormente citadas. Para ello se ha llevado a cabo la recopilación y análisis de la información hidrogeológica correspondiente a la documentación de los planes hidrológicos del Tajo (Plan Hidrológico de la cuenca del Tajo RD 1664/1998 y el actual Plan Hidrológico de la parte española de la Demarcación Hidrográfica del Tajo RD 270/2014) y de los estudios previos realizados por el organismo de cuenca y el Instituto Geológico y Minero de España (lGME) fundamentalmente. En relación con la MASb Tajuña-Montes Universales -cuya extensión supera la zona seleccionada- dichos estudios consideran su estructura geológica y distribución litológica, con intercalaciones impermeables que actúan como barreras, dividiendo a éstas en Subunidades e identificando las zonas de drenaje de sus respectivos acuíferos. También se ha considerado la documentación y estudios previos del Plan Hidrológico Nacional sobre las Unidades Hidrogeológicas compartidas entre ámbitos geográficos de diferentes planes hidrológicos. Se concluye que las divisorias hidrográficas de las cabeceras son sensiblemente coincidentes o abarcan las Subunidades Montes Universales meridionales, Priego, Cifuentes, Zaorejas, u Montes Universales septentrionales, que drenan hacia el Tajo/Guadiela (bien directamente, bien a través de afluentes como el Gallo, Ablanquejo, Cabrillas, Cuervo…), MASb Molina de Aragón, que drena al Tajo a través del río Gallo y MASb Sigüenza—Maranchón, que drena su parte correspondiente hacia el Tajo a través del Ablanquejo. Se descartan – salvo la pequeña salvedad del manantial de Cifuentes- las conexiones hidrogeológicas con otras MASb o Subunidades por lo que las cabeceras del Tajo y del Guadiela pueden considerarse como un Sistema independiente donde las precipitaciones no evaporadas escurren superficialmente o se infiltran y descargan hacia los embalses de Entrepeñas y Buendía. La cuantificación diaria y acumulada de los balances hídricos ha permitido calcular la evolución aproximada de las reservas de agua subterránea desde la fecha inicial. Originalmente los balances se realizaron de forma separada en las cabeceras del Tajo y del Guadiela, cuyos valores acumulados manifestaron una tendencia creciente en la primera y decreciente en la segunda. Dicha situación se equilibra cuando el balance se practica conjuntamente en ambas, apreciándose en la variación del volumen de agua subterránea una evolución acorde hidrológicamente con los ciclos de verano/invierno y periodos de sequía, manteniéndose sus valores medios a largo/medio plazo, poniendo en evidencia la existencia de interconexiones subterráneas entre ambas cuencas. El balance conjunto, agregando la cabecera del Tajuña (que también comparte los materiales permeables de la MASb Tajuña-Montes Universales) no reveló la existencia de nuevas interrelaciones hidrogeológicas que influyeran en los balances hídricos realizados Tajo/Guadiela, confirmando las conclusiones de los estudios hidrogeológicos anteriormente analizados. Se ha procedido a confrontar y validar los resultados obtenidos de la evolución de las reservas de agua subterránea mediante los siguientes procedimientos alternativos: - Cálculo de los parámetros de desagüe de la curva de agotamiento correspondiente al volumen de agua subterránea drenante hacia el Tajo/Guadiela. Éste se ha realizado a partir de las aportaciones mensuales entrantes en los embalses de Entrepeñas y Buendía durante los meses de junio, julio, agosto y septiembre, cuyos valores responden al perfil típico de descargas de un acuífero. A partir de éstos se ha determinado el volumen drenante correspondiente al primero de junio de cada año de la serie histórica considerada. - Determinación del caudal base por el método Wallingford y deducción de los volúmenes drenantes. Estimación de las recarga anuales - Cuantificación de la recarga anual por el método Sanz, Menéndez Pidal de Navascués y Távara. Se obtuvieron valores de recarga muy aproximados entre los calculados por los dos últimos procedimientos citados. Respecto a las reservas de agua subterránea almacenadas siguen una evolución semejante en todos los casos, lo que ha permitido considerar válidos los resultados conseguidos mediante balance hídrico. Confirmada su solidez, se han buscado correlaciones simples entre el volumen de las reservas subterráneas (como indicador estimativo del efecto de la infiltración) y los volúmenes procedentes de la fusión. La conclusión es que estos últimos no tienen un efecto determinante a escala anual sobre la infiltración,recarga y variación de los volúmenes de agua subterránea, frente al peso de otras variables (precipitación y evapotranspiración). No obstante se ha encontrado una buena correlación múltiple entre la recarga estimada y la precipitación eficaz (precipitación menos evapotranspiración) y fusión, que ha permitido cuantificar la contribución de esta última. Posteriormente se ha recurrido a la selección de los episodios más intensos de acumulación /fusión en las cabeceras del Tajo y Guadiela. Y se procedió a la comparación entre los resultados obtenidos por aplicación del modelo de simulación en los mismos periodos (normalmente de varios días de duración) con datos reales y con datos ficticios de temperatura que anularan o disminuyeran la presencia de nieve, apreciándose una gran sensibilidad del efecto de la temperatura sobre la evapotranspiración y estableciéndose nuevamente correlaciones lineales entre los volúmenes de fusión y el incremento de reservas subterráneas. Las mismas confirman el efecto “favorecedor” de la acumulación de agua en forma de nieve y su posterior licuación, sobre sobre la infiltración de agua en el suelo y almacenamiento subterráneo. Finalmente se establecieron varios escenarios climáticos (+1ºC; +3ºC; +1ºC y – 10% precipitación; y 3ºC – 10% precipitación) compatibles con las previsiones del IPCC para mediados y finales del presente siglo, determinándose mediante simulación ASTER los correspondientes valores de fusión. La correlación establecida a escala anual ha permitido evaluar el efecto de la disminución del volumen de fusión - en los diferentes escenarios – sobre la recarga, pronosticando un descenso de los caudales de estiaje y la desaparición del “efecto nieve” sobre la infiltración y recarga con un aumento de 3ºC de temperatura. Teniendo en cuenta las condiciones de representatividad de la zona elegida, resulta verosímil la extensión de las anteriores conclusiones a otras cabeceras fluviales enclavadas en áreas de media montaña situadas entre 1000 a 2000m y sus efectos aguas abajo.Water infiltration into the soil and groundwater recharge deep water in aquifers is slow relative to other hydrological phenomena. The wording of this thesis aims to contribute to the study of the influence that the storage of solid precipitation as snow cover and its eventual melting may have on this process in mid-mountain areas (1000 - 2,000 m) where very often the headwaters of the peninsular rivers are located. For this party analysis of the different variables involved has over a given time period and a particular geographical area, so that their methodology is empirical in nature. The extension of the period (2002/03 to 2010/11) has been conditioned by the availability of the values of some of its key variables, as were the water equivalent of the snow and flows from melting. These have been obtained as a result of the application of ASTER model, developed in the program Evaluation of Water Resources from the Innivation (ERHIN), calibrated - among others data of rainfall, temperature and flow from turn System Automatic Hydrological Information (SAIH). Both programs were implemented by the Administration in the different Water Boards and to undertakings for current basin, in which the author participated development of this thesis. As for the study area has proceeded at its option considering the possible areas of midmountain in the presence of snow outside hydrological meaningful and they were lithology consisting of permeable outcrops that did not prevent infiltration into the ground and forming aquifers of some significance. We were interested discretion in the Tagus basin, therefore the strategic nature of it, as currently supplying surplus to other basins deficit- as the representative value of its climate and terrain conditions in relation to other peninsular river basins . To do this we started from the headwaters identified by the ERHIN program for its implementation snow interest to the ASTER model and Ground Water Bodies MASb (before UUHH Hydrogeological Units) defined in hydrological plans. The intersection in the territory of both criteria led eventually to the Alto Tajo, in which both requirements are met. The section was finalized in the period between the headwaters of the Tagus and Guadiela rivers and reservoirs end Entrepeñas and Buendia respectively checking points for calibration performed in ASTER modeling. Much of it runs on carbonate rocks (limestones and dolomites of Jurassic and Cretaceous) related MASb of Tajuña -Montes Universal, Molina de Aragón and Sigüenza-Maranchón. The daily values of water reserves in the form of snow, evapotranspiration and flow from melting were obtained from the results of this model, proceeding to the calculation of infiltration water balance during the study period considered, taking into account values of precipitation, evapotranspiration and input flow. This has required the prior examination of the hydrogeological conditions of your required in order to know the possible underground interconnections that could alter the balance between the intervening variables aforementioned area. For this we have carried out the collection and analysis of hydrogeological information relevant documentation Tagus river management plans (Hydrological Plan Tajo Basin RD 1664/1998 and the current Hydrological Plan of the Spanish part of the River Basin Tagus RD 270/2014) and previous studies by the basin organization and the Geological Survey of Spain (IGME) mainly. Regarding the MASb Tajuña- Montes Universal - whose length exceeds the area selected - these studies consider its geological structure and lithology distribution with waterproof collations that act as barriers, dividing it into subunits and identifying areas draining their respective aquifers. It has also considered the documentation and previous studies of the National Hydrological Plan on shared among different geographical areas management plans Hydrogeological Units. We conclude that river dividing the headers are substantially coincident or covering Subunits southern Universal Montes, Priego Cifuentes, Zaorejas and northern Universal Mounts, which drain into the Tagus / Guadiela (either directly or through tributaries such as Gallo, Ablanquejo , whitecaps , Raven ...), MASb Molina de Aragón which drains through the Tajo del Gallo and MASb Sigüenza- Maranchón river that drains into the Tagus using the Ablanquejo . Discarded - except the small exception of spring Cifuentes -hydrogeological connections with other MASb or Subunits so the headwaters of the Tagus and Guadiela be considered as a separate system, where rainfall not evaporated runs on surface or infiltrates and eventually discharged into reservoirs Entrepeñas and Buendia. The daily and cumulative quantification of water balances allowed us to compute the approximate evolution of groundwater reserves from its initial date. Initially balances were performed separately in the headwaters of the Tagus and Guadiela, whose cumulative values showed an increasing trend in the first and decreasing in the second. This situation is balanced when the balance is practiced together in both , appreciating the change in volume of groundwater hydrological evolution commensurate with the cycles of summer / winter and drought periods , keeping their average long / medium term values and putting in shows the existence of underground interconnections between the two basins. The overall balance, adding header Tajuña (which also shares the permeable materials MASb Tajuña -Montes Universal ) did not reveal the existence of new hydrogeological interrelationships that influenced water balances made Tajo / Guadiela, confirming the findings of the hydrogeological studies previously analyzed. We proceeded to confront and validate the results of the evolution of groundwater reserves by the following alternative procedures: - Calculate the parameters drain depletion curve corresponding to the volume of groundwater draining into the Tajo / Guadiela. This has been made from monthly inflows in the reservoirs of Entrepeñas and Buendia during the months of June, July, August and September, whose values match the typical profile of an aquifer discharges. From these has been determined for the first of June each year of the time series considered drainage volume - Determination of base flow by Wallingford method and deduction of drainage volumes. Estimate of annual recharge - Quantification of the annual recharge by the method Sanz Menéndez Pidal of Navascués and Távara. Very approximate values recharge between calculated for the last two mentioned methods were obtained. Concerning groundwater reserves stored follow a similar pattern in all cases, allowing consider valid the results achieved through water balance. Confirmed its robustness, simple correlations were sought between the volume of groundwater reserves (as estimated indicator of the effect of infiltration) and volumes from the melting. The conclusion is that the latter do not have a decisive effect on the annual scale infiltration, recharge and variation in volumes of groundwater, against the weight of other variables (precipitation and evapotranspiration). However found a good multiple correlation between the estimated recharge and effective precipitation (precipitation minus evapotranspiration) and fusion, which allowed quantify the contribution of the latter. Subsequently it has resorted to the selection of the most intense episodes of accumulation / melting in the headwaters of the Tagus and Guadiela. And we proceeded to the comparison between the results obtained by application of the simulation model in the same periods (usually several days) with real data and fictitious temperature data to annul or decrease the presence of snow, appreciating a great sensitivity of the effect of temperature on evapotranspiration and establishing linear correlations between the volumes of melting and increased groundwater reserves again. They confirm the “flattering " effect of water accumulation as snow and subsequent liquefaction of the infiltration of water into the soil and underground storage. Finally various climate scenarios (+1ºC; +3ºC; +1ºC y – 10% precipitation; y 3ºC – 10% precipitation) were established consistent with IPCC projections for mid - to late - century, determined through simulation ASTER corresponding values of melting. The correlation established on an annual scale has allowed to evaluate the effect of decreasing the volume of melt - in different scenarios - on recharge, predicting a decline in low flows and the disappearance of "snow effect" on infiltration and recharge with an increase of 3°C temperature. Given the conditions of representativeness of the chosen area, plausible extension of the above findings to other landlocked headwaters in mid-mountain areas located between 1000 to 2000m and its downstream effects.

Evidence of a role for cyclic ADP-ribose in long-term synaptic depression in hippocampus

Ca2+ released from presynaptic and postsynaptic intracellular stores plays important roles in activity-dependent synaptic plasticity, including long-term depression (LTD) of synaptic strength. At Schaffer collateral–CA1 synapses in the hippocampus, presynaptic ryanodine receptor-gated stores appear to mobilize some of the Ca2+ necessary to induce LTD. Cyclic ADP-ribose (cADPR) has recently been proposed as an endogenous activator of ryanodine receptors in sea urchin eggs and several mammalian cell types. Here, we provide evidence that cADPR-mediated signaling pathways play a key role in inducing LTD. We show that biochemical production of cGMP increases cADPR concentration in hippocampal slices in vitro, and that blockade of cGMP-dependent protein kinase, cADPR receptors, or ryanodine-sensitive Ca2+ stores each prevent the induction of LTD at Schaffer collateral–CA1 synapses. A lack of effect of postsynaptic infusion of either cADPR antagonist indicates a probable presynaptic site of action.

Evidence for keratinocyte stem cells in vitro: Long term engraftment and persistence of transgene expression from retrovirus-transduced keratinocytes

Epidermis is renewed by a population of stem cells that have been defined in vivo by slow turnover, label retention, position in the epidermis, and enrichment in β1 integrin, and in vitro by clonogenic growth, prolonged serial passage, and rapid adherence to extracellular matrix. The goal of this study is to determine whether clonogenic cells with long-term growth potential in vitro persist in vivo and give rise to a fully differentiated epidermis. Human keratinocytes were genetically labeled in culture by transduction with a retrovirus encoding the lacZ gene and grafted to athymic mice. Analysis of the cultures before grafting showed that 21.1–27.8% of clonogenic cells with the capacity for >30 generations were successfully transduced. In vivo, β-galactosidase (β-gal) positive cells participated in the formation of a fully differentiated epithelium and were detected throughout the 40-week postgraft period, initially as loosely scattered clusters and later as distinct vertical columns. Viable cells recovered from excised grafts were seeded at clonal densities and 23.3–33.3% of the colonies thus formed were β-gal positive. In addition, no evidence of transgene inactivation was obtained: all keratinocyte colonies recovered from grafted tissue that were β-gal negative also lacked the lacZ transgene. These results show that cells with long-term growth properties in vitro do indeed persist in vivo and form a fully differentiated epidermis, thereby exhibiting the properties of stem cells.

Long-term potentiation and dual-component quantal signaling in the dentate gyrus

Long-term potentiation (LTP) of excitatory transmission is an important candidate cellular mechanism for the storage of memories in the mammalian brain. The subcellular phenomena that underlie the persistent increase in synaptic strength, however, are incompletely understood. A potentially powerful method to detect a presynaptic increase in glutamate release is to examine the effect of LTP induction on the rate at which the use-dependent blocker MK-801 attenuates successive N-methyl-d-aspartic acid (NMDA) receptor-mediated synaptic signals. This method, however, has given apparently contradictory results when applied in hippocampal CA1. The inconsistency could be explained if NMDA receptors were opened by glutamate not only released from local presynaptic terminals, but also diffusing from synapses on neighboring cells where LTP was not induced. Here we examine the effect of pairing-induced LTP on the MK-801 blocking rate in two afferent inputs to dentate granule cells. LTP in the medial perforant path is associated with a significant increase in the MK-801 blocking rate, implying a presynaptic increase in glutamate release probability. An enhanced MK-801 blocking rate is not seen, however, in the lateral perforant path. This result still could be compatible with a presynaptic contribution to LTP in the lateral perforant path if intersynaptic cross-talk occurred. In support of this hypothesis, we show that NMDA receptors consistently sense more quanta of glutamate than do α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors. In the medial perforant path, in contrast, there is no significant difference in the number of quanta mediated by the two receptors. These results support a presynaptic contribution to LTP and imply that differences in intersynaptic cross-talk can complicate the interpretation of experiments designed to detect changes in transmitter release.

Administration of helper-dependent adenoviral vectors and sequential delivery of different vector serotype for long-term liver-directed gene transfer in baboons

The efficiency of first-generation adenoviral vectors as gene delivery tools is often limited by the short duration of transgene expression, which can be related to immune responses and to toxic effects of viral proteins. In addition, readministration is usually ineffective unless the animals are immunocompromised or a different adenovirus serotype is used. Recently, adenoviral vectors devoid of all viral coding sequences (helper-dependent or gutless vectors) have been developed to avoid expression of viral proteins. In mice, liver-directed gene transfer with AdSTK109, a helper-dependent adenoviral (Ad) vector containing the human α1-antitrypsin (hAAT) gene, resulted in sustained expression for longer than 10 months with negligible toxicity to the liver. In the present report, we have examined the duration of expression of AdSTK109 in the liver of baboons and compared it to first-generation vectors expressing hAAT. Transgene expression was limited to approximately 3–5 months with the first-generation vectors. In contrast, administration of AdSTK109 resulted in transgene expression for longer than a year in two of three baboons. We have also investigated the feasibility of circumventing the humoral response to the virus by sequential administration of vectors of different serotypes. We found that the ineffectiveness of readministration due to the humoral response to an Ad5 first-generation vector was overcome by use of an Ad2-based vector expressing hAAT. These data suggest that long-term expression of transgenes should be possible by combining the reduced immunogenicity and toxicity of helper-dependent vectors with sequential delivery of vectors of different serotypes.

CTLA4-Ig and anti-CD40 ligand prevent renal allograft rejection in primates

Selective inhibition of T cell costimulation using the B7-specific fusion protein CTLA4-Ig has been shown to induce long-term allograft survival in rodents. Antibodies preventing the interaction between CD40 and its T cell-based ligand CD154 (CD40L) have been shown in rodents to act synergistically with CTLA4-Ig. It has thus been hypothesized that these agents might be capable of inducing long-term acceptance of allografted tissues in primates. To test this hypothesis in a relevant preclinical model, CTLA4-Ig and the CD40L-specific monoclonal antibody 5C8 were tested in rhesus monkeys. Both agents effectively inhibited rhesus mixed lymphocyte reactions, but the combination was 100 times more effective than either drug alone. Renal allografts were transplanted into nephectomized rhesus monkeys shown to be disparate at major histocompatibility complex class I and class II loci. Control animals rejected in 5–8 days. Brief induction doses of CTLA4-Ig or 5C8 alone significantly prolonged rejection-free survival (20–98 days). Two of four animals treated with both agents experienced extended (>150 days) rejection-free allograft survival. Two animals treated with 5C8 alone and one animal treated with both 5C8 and CTLA4-Ig experienced late, biopsy-proven rejection, but a repeat course of their induction regimen successfully restored normal graft function. Neither drug affected peripheral T cell or B cell counts. There were no clinically evident side effects or rejections during treatment. We conclude that CTLA4-Ig and 5C8 can both prevent and reverse acute allograft rejection, significantly prolonging the survival of major histocompatibility complex-mismatched renal allografts in primates without the need for chronic immunosuppression.

Physiological response to long-term peripheral and central leptin infusion in lean and obese mice

Recent data have identified leptin as an afferent signal in a negative-feedback loop regulating the mass of the adipose tissue. High leptin levels are observed in obese humans and rodents, suggesting that, in some cases, obesity is the result of leptin insensitivity. This hypothesis was tested by comparing the response to peripherally and centrally administered leptin among lean and three obese strains of mice: diet-induced obese AKR/J, New Zealand Obese (NZO), and Ay. Subcutaneous leptin infusion to lean mice resulted in a dose-dependent loss of body weight at physiologic plasma levels. Chronic infusions of leptin intracerebroventricularly (i.c.v.) at doses of 3 ng/hr or greater resulted in complete depletion of visible adipose tissue, which was maintained throughout 30 days of continuous i.c.v. infusion. Direct measurement of energy balance indicated that leptin treatment did not increase total energy expenditure but prevented the decrease that follows reduced food intake. Diet-induced obese mice lost weight in response to peripheral leptin but were less sensitive than lean mice. NZO mice were unresponsive to peripheral leptin but were responsive to i.c.v. leptin. Ay mice did not respond to subcutaneous leptin and were 1/100 as sensitive to i.c.v. leptin. The decreased response to leptin in diet-induced obese, NZO, and Ay mice suggests that obesity in these strains is the result of leptin resistance. In NZO mice, leptin resistance may be the result of decreased transport of leptin into the cerebrospinal fluid, whereas in Ay mice, leptin resistance probably results from defects downstream of the leptin receptor in the hypothalamus.

Glycine-induced long-term potentiation is associated with structural and functional modifications of α-amino-3-hydroxyl-5-methyl-4-isoxazolepropionic acid receptors

Global long-term potentiation (LTP) was induced in organotypic hippocampal slice cultures by a brief application of 10 mM glycine. Glycine-induced LTP was occluded by previous theta burst stimulation-induced potentiation, indicating that both phenomena share similar cellular processes. Glycine-induced LTP was associated with increased [3H]α-amino-3-hydroxyl-5-methyl-4-isoxazolepropionic acid (AMPA) binding in membrane fractions as well as increased amount of a selective spectrin breakdown product generated by calpain-mediated spectrin proteolysis. Antibodies against the C-terminal (C-Ab) and N-terminal (N-Ab) domains of GluR1 subunits were used to evaluate structural changes in AMPA receptor properties resulting from glycine-induced LTP. No quantitative or qualitative changes were observed in Western blots from membrane fractions prepared from glycine-treated slices with C-Ab. In contrast, Western blots stained with N-Ab revealed the formation of a 98-kDa species of GluR1 subunits as well as an increased amount of immunoreactivity after glycine-induced LTP. The amount of spectrin breakdown product was positively correlated with the amount of the 98-kDa species of GluR1 after glycine treatment. Functional modifications of AMPA receptors were evaluated by determining changes in the effect of pressure-applied AMPA on synaptic responses before and after glycine-induced LTP. Glycine treatment produced a significant increase in AMPA receptor function after potentiation that correlated with the degree of potentiation. The results indicate that LTP induction produces calpain activation, truncation of the C-Ab domain of GluR1 subunits of AMPA receptors, and increased AMPA receptor function. They also suggest that insertion of new receptors takes place after LTP induction.

Paracrine stimulation of interstitial collagenase (MMP-1) in the human endometrium by interleukin 1α and its dual block by ovarian steroids

In the cycling human endometrium, the expression of interstitial collagenase (MMP-1) and of several related matrix metalloproteinases (MMPs) follows the late-secretory fall in sex steroid plasma concentrations and is thought to be a critical step leading to menstruation. The rapid and extensive lysis of interstitial matrix that precedes menstrual shedding requires a strict control of these proteinases. However, the mechanism by which ovarian steroids regulate endometrial MMPs remains unclear. We report here that, in the absence of ovarian steroids, MMP-1 expression in endometrial fibroblasts is markedly stimulated by medium conditioned by endometrial epithelial cells. This stimulation can be prevented by antibodies directed against interleukin 1α (IL-1α) but not against several other cytokines. Ovarian steroids inhibit the release of IL-1α and repress MMP-1 production by IL-1α-stimulated fibroblasts. In short-term cultures of endometrial explants obtained throughout the menstrual cycle, the release of both IL-1α and MMP-1 is essentially limited to the perimenstrual phase. We conclude that epithelium-derived IL-1α is the key paracrine inducer of MMP-1 in endometrial fibroblasts. However, MMP-1 production in the human endometrium is ultimately blocked by ovarian steroids, which act both upstream and downstream of IL-1α, thereby exerting an effective control via a “double-block” mechanism.

Impaired long-term potentiation induction in dentate gyrus of calretinin-deficient mice

Calretinin (Cr) is a Ca2+ binding protein present in various populations of neurons distributed in the central and peripheral nervous systems. We have generated Cr-deficient (Cr−/−) mice by gene targeting and have investigated the associated phenotype. Cr−/− mice were viable, and a large number of morphological, biochemical, and behavioral parameters were found unaffected. In the normal mouse hippocampus, Cr is expressed in a widely distributed subset of GABAergic interneurons and in hilar mossy cells of the dentate gyrus. Because both types of cells are part of local pathways innervating dentate granule cells and/or pyramidal neurons, we have explored in Cr−/− mice the synaptic transmission between the perforant pathway and granule cells and at the Schaffer commissural input to CA1 pyramidal neurons. Cr−/− mice showed no alteration in basal synaptic transmission, but long-term potentiation (LTP) was impaired in the dentate gyrus. Normal LTP could be restored in the presence of the GABAA receptor antagonist bicuculline, suggesting that in Cr−/− dentate gyrus an excess of γ-aminobutyric acid (GABA) release interferes with LTP induction. Synaptic transmission and LTP were normal in CA1 area, which contains only few Cr-positive GABAergic interneurons. Cr−/− mice performed normally in spatial memory task. These results suggest that expression of Cr contributes to the control of synaptic plasticity in mouse dentate gyrus by indirectly regulating the activity of GABAergic interneurons, and that Cr−/− mice represent a useful tool to understand the role of dentate LTP in learning and memory.

Synergistic interaction between leptin and cholecystokinin to reduce short-term food intake in lean mice

Leptin is a circulating protein involved in the long-term regulation of food intake and body weight. Cholecystokinin (CCK) is released postprandially and elicits satiety signals. We investigated the interaction between leptin and CCK-8 in the short-term regulation of food intake induced by 24-hr fasting in lean mice. Leptin, injected intraperitoneally (i.p.) at low doses (4–120 μg/kg), which did not influence feeding behavior for the first 3 hr postinjection, decreased food intake dose dependently by 47–83% during the first hour when coinjected with a subthreshold dose of CCK. Such an interaction was not observed between leptin and bombesin. The food-reducing effect of leptin injected with CCK was not associated with alterations in gastric emptying or locomotor behavior. Leptin–CCK action was blocked by systemic capsaicin at a dose inducing functional ablation of sensory afferent fibers and by devazepide, a CCK-A receptor antagonist but not by the CCK-B receptor antagonist, L-365,260. The decrease in food intake which occurs 5 hr after i.p. injection of leptin alone was also blunted by devazepide. Coinjection of leptin and CCK enhanced the number of Fos-positive cells in the hypothalamic paraventricular nucleus by 60%, whereas leptin or CCK alone did not modify Fos expression. These results indicate the existence of a functional synergistic interaction between leptin and CCK leading to early suppression of food intake which involves CCK-A receptors and capsaicin-sensitive afferent fibers.

The exaptive excellence of spandrels as a term and prototype

In 1979, Lewontin and I borrowed the architectural term “spandrel” (using the pendentives of San Marco in Venice as an example) to designate the class of forms and spaces that arise as necessary byproducts of another decision in design, and not as adaptations for direct utility in themselves. This proposal has generated a large literature featuring two critiques: (i) the terminological claim that the spandrels of San Marco are not true spandrels at all and (ii) the conceptual claim that they are adaptations and not byproducts. The features of the San Marco pendentives that we explicitly defined as spandrel-properties—their necessary number (four) and shape (roughly triangular)—are inevitable architectural byproducts, whatever the structural attributes of the pendentives themselves. The term spandrel may be extended from its particular architectural use for two-dimensional byproducts to the generality of “spaces left over,” a definition that properly includes the San Marco pendentives. Evolutionary biology needs such an explicit term for features arising as byproducts, rather than adaptations, whatever their subsequent exaptive utility. The concept of biological spandrels—including the examples here given of masculinized genitalia in female hyenas, exaptive use of an umbilicus as a brooding chamber by snails, the shoulder hump of the giant Irish deer, and several key features of human mentality—anchors the critique of overreliance upon adaptive scenarios in evolutionary explanation. Causes of historical origin must always be separated from current utilities; their conflation has seriously hampered the evolutionary analysis of form in the history of life.

Interleukin 12 and B7-1 costimulatory molecule expressed by an adenovirus vector act synergistically to facilitate tumor regression

Stimulation of antitumor immune mechanisms is the primary goal of cancer immunotherapy, and accumulating evidence suggests that effective alteration of the host–tumor relationship involves immunomodulating cytokines and also the presence of costimulatory molecules. To examine the antitumor effect of direct in vivo gene transfer of murine interleukin 12 (IL-12) and B7-1 into tumors, we developed an adenovirus (Ad) vector, AdIL12–B7-1, that encodes the two IL-12 subunits in early region 1 (E1) and the B7-1 gene in E3 under control of the murine cytomegalovirus promoter. This vector expressed high levels of IL-12 and B7-1 in infected murine and human cell lines and in primary murine tumor cells. In mice bearing tumors derived from a transgenic mouse mammary adenocarcinoma, a single intratumoral injection with a low dose (2.5 × 107 pfu/mouse) of AdIL12–B7-1 mediated complete regression in 70% of treated animals. By contrast, administration of a similar dose of recombinant virus encoding IL-12 or B7-1 alone resulted in only a delay in tumor growth. Interestingly, coinjection of two different viruses expressing either IL-12 or B7-1 induced complete tumor regression in only 30% of animals treated at this dose. Significantly, cured animals remained tumor free after rechallenge with fresh tumor cells, suggesting that protective immunity had been induced by treatment with AdIL12–B7-1. These results support the use of Ad vectors as a highly efficient delivery system for synergistically acting molecules and show that the combination of IL-12 and B7-1 within a single Ad vector might be a promising approach for in vivo cancer therapy.