Morphological plasticity of the coral skeleton under CO2-driven seawater acidification

Ocean acidification causes corals to calcify at reduced rates, but current understanding of the underlying processes is limited. Here, we conduct a mechanistic study into how seawater acidification alters skeletal growth of the coral Stylophora pistillata. Reductions in colony calcification rates are manifested as increases in skeletal porosity at lower pH, while linear extension of skeletons remains unchanged. Inspection of the microstructure of skeletons and measurements of pH at the site of calcification indicate that dissolution is not responsible for changes in skeletal porosity. Instead, changes occur by enlargement of corallite-calyxes and thinning of associated skeletal elements, constituting a modification in skeleton architecture. We also detect increases in the organic matrix protein content of skeletons formed under lower pH. Overall, our study reveals that seawater acidification not only causes decreases in calcification, but can also cause morphological change of the coral skeleton to a more porous and potentially fragile phenotype.

Atmospheric electric measurement in the troposphere and stratosphere on the Atlantic Ocean during 1965 and 1969

One main point of the air electric investigations at the atlantic 1965 and 1969 was the record of the potential gradient in the troposphere with free and captive balloon ascents. The course of the field vs. altitude above the sea differs from that over land. A remarkable enlargement of the field strength occurs at the altitude of the passat inversion. The electric voltage between ionosphere and earth could be obtained by integrating the potential gradient over the altitude. Such computations have been made by balloon ascents simultaneous over the ocean and at Weissenau (South Germany), From 15 simultaneous measurements the average value of the potential of the ionosphere over the ocean is 214 kV and over South Germany 216 kV, that means very close together. Because of the small differences also between the single values it can be concluded that in generally the ionosphere potential has an equal value over these both places at one moment. From the potential of the ionosphere VI, the field strength E0 and the conductivity lamda o, both measured at the sea surface, the columnar resistance R could be derived to 2.4 x 10**17 Ohm x m**2. By correlation of the single values of the ionosphere potential with the potential gradient measured simultaneously at the surface of the sea a linear proportional relationship exists; it follows from this result, that R is nearly constant. The mean value of the air-earth current density over the ocean could be calculated by using the measured values of the small ion density with respect to the electrode effect prooved at the equator station. The current density was only 0.9 x 10**-12 A/m**2, which means, a three and a half times smaller value than estimated by Carnegie and accepted up to now. Therefore it seems to be necessary to correct the former calculations of the global current balance.

Preliminary Study and Proposal for Restoring a Jesuit College at Saragossa, Spain

A historical study and a construction pathology survey were conducted prior to proposing a solution for restoring the upper windows on the south side of Old San Carlos College at Saragossa (16th-17th centuries), wich had been forteited to add a third storey to the cloister. Althought initially designed to a simple large hall format, the church is a harmonius blend of Aragonese Gothic architecture and typical Jesuit scheme, consisting in a central nave flanket by chapel-confessional and raised galleries for the community. A subsequent enlargement of the roof, wich rests on the original framing over the central nave, reduced the mechanical strength of the principal rafters on the opposite side, prompting a concomitant imbalance of forces that has affected the entire structure. In view of the foregoing, in addition to restituting the upper window, the proposal solution envisages restoring the roof over the central nave to its original design, and with the interior lighting in the church.

Container Terminal Simulation. A case study: port of Valencia.

This paper is concerned with the study of non-Markovian queuing systems in container terminals. The methodology presented has been applied to analyze the ship traffic in the port of Valencia located in the Western Mediterranean. Two container terminals have been studied: the public container terminal of NOATUM and the dedicated container terminal of MSC. This paper contains the results of a simulation model based on queuing theory. The methodology presented is found to be effective in replicating realistic ship traffic operations in port as well as in conducting capacity evaluations. Thus the methodology can be used for capacity planning (long term), tactical planning (medium term) and even for the container terminal design (port enlargement purposes).

La rehabilitación de construcciones militares para uso hotelero: la red de Paradores de Turismo, (1928-2012)

Fundamento de la tesis: Al comienzo del siglo XX, el interés por el turismo unido a la necesidad de restaurar un abundante patrimonio histórico, posibilitó en España que los poderes públicos se embarcaran en una singular experiencia: la creación de una infraestructura hotelera a partir de la rehabilitación de edificios históricos. La preservación, mantenimiento e incluso rentabilidad de una gran parte del patrimonio español se haría efectiva a través de la innovadora fórmula patrimonio‐turismo, cuya máxima expresión se materializó en la Red de Paradores desde su fundación en la segunda década del siglo pasado hasta nuestros días. Sorprendentemente, este tema no ha sido todavía investigado en su vertiente arquitectónica pese a que España ha sido pionera y modelo en la cuestión de la hotelería pública. Este trabajo aborda el estudio del caso más significativo de todos los edificios de la red, en tanto que el patrimonio que ha servido de base a los fines hoteleros del Estado ha contado con un total de seis tipos arquitectónicos a lo largo de su historia, dentro de los cuales la arquitectura militar despunta con su mayoritaria presencia dentro del contexto de los edificios históricos de la red. El carácter arquetípico de los castillos y fortalezas, arraigado en el inconsciente colectivo, les hizo especialmente atractivos como alojamiento turístico al permitir evocar la remota época medieval, pese a ser el tipo arquitectónico más comprometido para la rehabilitación hotelera. El estudio de las intervenciones operadas en estos inmuebles se revela de forma clara como escaparate de los distintos criterios de intervención patrimonial que se han sucedido en el siglo XX, hasta enlazar con la perspectiva interdisciplinar actual. La tesis abarca en, primer lugar, diferentes aspectos generales relativos al promotor hotelero, la hotelería pública de ámbito nacional e internacional, y la caracterización de los inmuebles de la red estatal española, desde el punto de vista hotelero y arquitectónico, entendida esta última en sus tres escalas de influencia: la arquitectónica, la urbana o paisajística, y la del interiorismo. Se analiza en segundo término la arquitectura militar dentro del contexto de la Red de Paradores, desde la consideración de su transformación hotelera, para lo cual ha sido necesario realizar una clasificación propia, que abarca tanto edificios que respondieron a una estructura de cuartel, como castillos‐palacio, o fortalezas que habían servido a los fines de una orden religiosa militar, además de considerarse las intervenciones en recintos históricos de carácter militar, donde se hacía obligatorio construir de nueva planta. En tercer y último lugar, se analiza a lo largo de las distintas etapas del organismo turístico las rehabilitaciones realizadas en estas construcciones militares, a la vez que se tienen en cuenta las intervenciones en los restantes edificios históricos, para evitar la descontextualización. Este recorrido comienza con la promoción de los dos primeros paradores a cargo del Comisario Regio, el marqués de la Vega‐Inclán, que sirvieron para sentar las bases de los conceptos e ideas que habrían de desarrollarse en las siguientes décadas. Posteriormente, se desarrolló y tomó forma la red con el Patronato Nacional del Turismo, en la que las primeras intervenciones en tipos militares se tradujeron en reformas interiores de locales. La etapa clave de la red, y en particular de la arquitectura militar, tuvo lugar con el Ministerio de Información y Turismo, marcada por la “repristinación” de monumentos, tras un período preparatorio con la Dirección General del Turismo en el que lo militar había quedado de telón de fondo de otros tipos arquitectónicos. Tras el auge del Ministerio llegó el período de decadencia en el que los castillos y fortalezas desaparecieron de los intereses de las Secretarias de Turismo, hasta llegar a las inauguraciones de los novedosos establecimientos del siglo XXI y el resurgimiento del tipo militar con el parador de Lorca. Metodología empleada: Este trabajo de investigación se ha servido fundamentalmente de documentación inédita, procedente de diversos archivos, además de una muy extensa toma de datos in situ. Dentro del patrimonio analizado, los inmuebles que responden al tipo arquitectónico militar se han dividido en tres grandes grupos: inmuebles rehabilitados que entraron en funcionamiento en la red, inmuebles en proceso de transformación hotelera, e inmuebles que fueron adquiridos con fines hoteleros pero que no llegaron a rehabilitarse. Para cada uno de ellos ha sido necesario determinar en qué estado llegaron a manos de la Administración Turística, cuál fue el mecanismo a través del cual se adquirieron, en qué consistió su primera rehabilitación hotelera, y cuáles fueron las ampliaciones o reformas más significativas que se realizaron posteriormente. Estos datos se han sintetizado en fichas y se han extraído conclusiones al comparar cada unidad con el conjunto. Simultáneamente se introdujeron dos factores externos: la historia del turismo que permitió hacer una ordenación cronológica de los inmuebles según etapas, y la historia de la teoría y práctica de la intervención patrimonial en España que permitió comparar los criterios patrimoniales de la Administración competente respecto de las intervenciones de la Administración Turística, cuyo contacto se haría obligatorio a partir del Decreto, de 22 de abril de 1949, que dejaba bajo la tutela del Estado a todos los castillos y fortalezas. Aportación de la tesis: Con carácter general, la tesis centra una ordenación y sistematización completa del patrimonio inmobiliario de la red, desde el punto de vista de los tipos hoteleros y arquitectónicos, además de poner por primera vez en conexión distintos modelos de hotelería pública, para constituirse en el sustrato de futuras investigaciones. El estudio realizado se ha hecho extensivo a las distintas escalas que inciden de forma interconectada en la implantación de un parador: la arquitectónica, la urbana y la del interiorismo, hasta ahora referenciado desde la exclusiva visión arquitectónica. Se han definido las etapas de la historia de la red, no ya sólo a partir del hilo conductor de la cadena sucesiva de organismos turísticos, sino que por primera vez se hace en razón de la evolución que sufren las intervenciones patrimoniales a lo largo del tiempo, a la vez que se entra en conexión con la teoría y praxis de la restauración monumental. Con carácter particular, la arquitectura militar dentro del contexto de los paradores se destaca en el período del Ministerio, en el que se experimentaron todas las posibilidades que presentaba su rehabilitación. En este sentido se ha puesto de manifiesto en este trabajo un tipo híbrido de parador, a caballo entre la rehabilitación y la edificación de nueva planta, las dos formas básicas de establecimiento creadas en la Comisaría Regia, al que se ha denominado edificación de nueva planta en recinto histórico militar. Esta nueva caracterización se ha valorado como la forma más eficiente de implantar paradores, cuyas pautas arquitectónicas abarcaron un abanico de posibilidades: imitación de modelos arquitectónicos históricos con utilización de elementos patrimoniales prestados que dieran el valor de la historia, utilización de un lenguaje moderno, o la inspiración en la arquitectura vernácula. La amalgama de elementos, estilos e intervenciones sucesivas de ampliación fue la característica común tanto para la implantación de un parador en un edificio como en un recinto amurallado. La arquitectura militar transformada en establecimiento hotelero evidencia la vocación escenográfica de las intervenciones patrimoniales, secundada por el interiorismo, además de su aportación a la arquitectura hotelera en lo referente al confort, organización y funcionamiento de sus instalaciones. La tesis ahonda en los diversos aspectos de la rehabilitación hotelera apuntados de forma parcial por algunos autores, y pone de manifiesto la “ambientación medieval” operada en la arquitectura militar, que llegó a tener su máxima expresión con el criterio de la “unidad de estilo” del Ministerio de Información y Turismo. La rehabilitación hotelera dentro del contexto de la Red de Paradores, queda caracterizada en la tesis en relación a intervenciones en construcciones militares, cuya sistematización puede ser extrapolable a otros tipos arquitectónicos o cadenas hoteleras de titularidad pública, a partir del estudio que se ha avanzado en este trabajo. Thesis basis: At the beginning of the 20th century the interest in tourism added to the plentiful heritage in Spain enabled the authorities to embark on a singular experience: the creation of a hotel infrastructure from the restoration of historic buildings. Preservation, maintenance, and even profitability of a large part of the Spanish heritage would be effective through the innovative formula heritage-tourism. Its greatest expression materialized in the Paradores Network since its foundation in last century’s second decade to the present day. Surprisingly, this subject has not yet been investigated in its architectural aspect, even though Spain has been a pioneer and a model in the matter of public hotel business. This project tackles the study of the most significative case of all the network’s buildings, since the heritage which has served throughout history as a base for the State hotel purposes has altogether six architectural types, among which military architecture stands out with its majority presence in the context of the historical buildings of the network. The archetypal character of castles and fortresses, ingrained in the collective subconscious, made them specially attractive for tourist accommodation, as it allowed the evocation of far medieval times, despite being the most awkward architectural type for hotel restoration. The study of the interventions in these buildings clearly reveals itself as a showcase of the different criteria of heritage intervention along the 20th century, connecting to the present interdisciplinary perspective. Firstly, the thesis covers different general aspects regarding the hotel developer, the domestic and international public hotel business, and the description of the Spanish state network buildings from a hotel business and an architectural point of view, the latter from its three influence scales: architectural, urban or landscape, and interior design. Secondly, the transformation of the military architecture in the Paradores Network into hotels is analyzed. For that purpose it was necessary to create a specific classification, which included barrack-structured buildings, castle-palaces, or fortresses which served the purposes of military-religious orders. The interventions in those military historical places where new building became compulsory were also taken into consideration. Thirdly and lastly, the thesis analyses the restorations in these military constructions through the different stages of the tourist organization. In order to avoid decontextualization, interventions in other historical buildings were also considered. This route begins with the promotion of the two first Paradores by the Royal Commissioner, the marquis of Vega-Inclán, which paved the way for the concepts and ideas that were developed in the following decades. Subsequently, the network was developed and took shape with the National Tourism Board. The first interventions on military types were inside refurbishments. The Network’s key period, and in particular of its military architecture, took place with the Ministry of Information and Tourism, a time marked by the “restoration to its original state” of monuments. This stage arrived after a preparatory period with the State Tourist Office, when the military type was left as a backdrop for other architectural types. After the Ministry’s boom arrived a decline, in which castles and fortresses disappeared from the Tourist Department’s interests up to the opening of the 21st century new establishments and the resurgence of the military type with Lorca’s Parador. Methodology: The present research project has mainly used unpublished documentation from several archives and has done an extensive in situ data-gathering. Within the heritage analyzed, military buildings have been divided into three main groups: restored buildings that began to operate in the network, those in process of hotel transformation, and those acquired for hotel purposes, but which did not become restored. In each case, it has been necessary to determine the condition in which they arrived to the Tourist Administration, the procedure by which they were acquired, what their first hotel restoration consisted of, and which their subsequent most significative enlargements and alterations were. These facts have been synthesized in cards, and conclusions were drawn by comparing each unit with the whole. Simultaneously, two external factors were introduced: the history of tourism, that allowed establishing a chronological order according to different periods, and the history of Spanish heritage intervention’s theory and practice, that permitted to compare the heritage criteria from the competent Administration with those of the Tourist Administration’s interventions. Both Administrations came compulsorily into contact after the Decree of 22nd April 1949, by which all castles and fortresses became under the protection of the State. Thesis contribution: In general, the thesis focuses on a complete order and systematization of the network’s heritage buildings from the hotel and architectural types points of view, besides connecting for the first time different public hotel business models, becoming the substratum for future investigations. The study has included the different scales that impact interconnected on the establishment of a Parador: architectural, urban and interior design, only referenced to date from an architectural point of view. The Network’s history stages have been defined according to not only a consecutive series of tourist organizations, but also, and for the first time, to the evolution of heritage interventions over time, thus connecting with the theory and praxis of monumental restoration. In particular, within the Paradores, military architecture stands out in the Ministry’s period, in which all kind of restoration possibilities were explored. In this sense, the present project puts forth a hybrid type of Parador between restoration and new building, the two basic ways of establishment created in the Royal Commission, termed new building in military historic enclosure. This new characterization has been evaluated as the most efficient for establishing Paradores, whose architectonic guidelines include a wide range of possibilities: the imitation of historical architectonic models with use of borrowed heritage components that provide historical value, the use of modern language, or the inspiration in vernacular architecture. The amalgam of elements, styles and consecutive enlargement interventions was the common feature of the establishment of a Parador, both in a building or in a walled enclosure. The military architecture transformed into a hotel establishment gives proof of the scenographic vocation of heritage interventions, supported by interior design, as well as of its contribution to hotel architecture, related to its comfort, organization and the functioning of its facilities. The thesis delves into the diverse aspects of hotel restoration, partially pointed out by several authors, and puts forth the creation of a “medieval atmosphere” in military architecture, which came to its highest expression with the “unitary style” criteria of the Ministry of Information and Tourism. Hotel restoration within the context of the Paradores’ Network is defined in this thesis in relation to interventions in military constructions, whose systemization can be extrapolative to other architectural types or public hotel chains, based on the study which has been put forward in this project.

Modelo Híbrido para la Ayuda al Capitán

En esta Tesis se plantea una nueva forma de entender la evacuación apoyándonos en tecnologías existentes y accesibles que nos permitirán ver este proceso como un ente dinámico. Se trata de una metodología que implica no solo el uso de herramientas de análisis que permitan la definición de planes de evacuación en tiempo real, sino que también se apunta hacia la creación de una infraestructura física que permita alimentar con información actualizada al sistema de forma que, según la situación y la evolución de la emergencia, sea posible realizar planes alternativos que se adapten a las nuevas circunstancias. En base a esto, el sistema asimilará toda esa información y aportará soluciones que faciliten la toma de decisiones durante toda la evolución del incidente. Las aportaciones originales de esta Tesis son múltiples y muy variadas, pudiéndolas resumir en los siguientes puntos: 1. Estudio completo del estado del arte: a. Detección y análisis de diferentes proyectos a nivel internacional que de forma parcial tratan algunos aspectos desarrollados en la Tesis. b. Completo estudio a nivel mundial del software desarrollado total o parcialmente para la simulación del comportamiento humano y análisis de procesos de evacuación. Se ha generado una base de datos que cataloga de forma exhaustiva estas aplicaciones, permitiendo realizar un completo análisis y posibilitando la evolución futura de los contenidos de la misma. En la tesis se han analizado casi un centenar de desarrollos, pero el objetivo es seguir completando esta base de datos debido a la gran utilidad y a las importantes posibilidades que ofrece. 2. Desarrollo de un importante capítulo que trata sobre la posibilidad de utilizar entornos virtuales como alternativa intermedia al uso de simuladores y simulacros. En esta sección se divide en dos bloques: a. Ensayos en entornos reales y virtuales. b. Ensayos en entornos virtuales (pruebas realizadas con varios entornos virtuales). 3. Desarrollo de e-Flow net design: paquete de herramientas desarrolladas sobre Rhinoceros para el diseño de la red de evacuación basada en los elementos definidos en la tesis: Nodes, paths, Relations y Areas. 4. Desarrollo de e-Flow Simulator: Conjunto de herramientas que transforman Rhinoceros en un simulador 3D de comportamiento humano. Este simulador, de desarrollo propio, incorpora un novedoso algoritmo de comportamiento a nivel de individuo que incluye aspectos que no se han encontrado en otros simuladores. Esta herramienta permite realizar simulaciones programadas de grupos de individuos cuyo comportamiento se basa en el análisis del entorno y en la presencia de referencias dinámicas. Incluye otras importantes novedades como por ejemplo: herramientas para análisis de la señalización, elementos de señalización dinámica, incorporación sencilla de obstáculos, etc. También se ha creado una herramienta que posibilita la implementación del movimiento del propio escenario simulando la oscilación del mismo, con objeto de reflejar la influencia del movimiento del buque en el desplazamiento de los individuos. 5. En una fase avanzada del desarrollo, se incorporó la posibilidad de generar un vídeo de toda la simulación, momento a partir del cual, se han documentado todas las pruebas (y se continúan documentando) en una base de datos que recoge todas las características de las simulaciones, los problemas detectados, etc. Estas pruebas constituyen, en el momento en que se ha cerrado la redacción de la Tesis, un total de 81 GB de datos. Generación y análisis de rutas en base a la red de evacuación creada con e-Flow Net design y las simulaciones realizadas con e-Flow Net simulator. a. Análisis para la optimización de la configuración de la red en base a los nodos por área existentes. b. Definición de procesos previos al cálculo de rutas posibles. c. Cálculo de rutas: i. Análisis de los diferentes algoritmos que existen en la actualidad para la optimización de rutas. ii. Desarrollo de una nueva familia de algoritmos que he denominado “Minimum Decision Algorithm (MDA)”, siendo los algoritmos que componen esta familia: 1. MDA básico. 2. MDA mínimo. 3. MDA de no interferencia espacial. 4. MDA de expansión. 5. MDA de expansión ordenada para un único origen. 6. MDA de expansión ordenada. iii. Todos estos algoritmos se han implementado en la aplicación e-Flow creada en la Tesis para el análisis de rutas y que constituye el núcleo del Sistema de Ayuda al Capitán. d. Determinación de las alternativas para el plan de evacuación. Tras la definición de las rutas posibles, se describen diferentes procesos existentes de análisis por ponderación en base a criterios, para pasar finalmente a definir el método de desarrollo propio propuesto en esta Tesis y cuyo objetivo es responder en base a la población de rutas posibles obtenidas mediante los algoritmos MDA, qué combinación de rutas constituyen el Plan o Planes más adecuados para cada situación. La metodología creada para la selección de combinaciones de rutas que determinan un Plan completo, se basa en cuatro criterios básicos que tras su aplicación ofrecen las mejores alternativas. En esta fase también se incluye un complejo análisis de evolución temporal que incorpora novedosas definiciones y formulaciones. e. Derivado de la definición de la metodología creada en esta Tesis para la realización de los análisis de evolución temporal, se ha podido definir un nuevo teorema matemático que se ha bautizado como “Familia de cuadriláteros de área constante”. 7. Especificación de la infraestructura física del Sistema de Ayuda al Capitán: parte fundamental de sistema es la infraestructura física sobre la que se sustentaría. Esta infraestructura estaría compuesta por sensores, actuadores, aplicaciones para dispositivos móviles, etc. En este capítulo se analizan los diferentes elementos que la constituirían y las tecnologías implicadas. 8. Especificación de la infraestructura de servicios. 9. Creación del Blog Virtual Environments (http://epcinnova-virtualenvironments.blogspot.com.es/) en el que se han publicado todas las pruebas realizadas en el capítulo que analiza los entornos virtuales como alternativa a los simuladores y a los ensayos en laboratorio o los simulacros, incluyendo en muchos casos la posibilidad de que el visitante del blog pueda realizar la simulación en el entorno virtual. Este blog también incluye otras secciones que se han trabajado durante la Tesis: • Recopilación de diferentes entornos virtuales existentes. • Diagrama que recopila información sobre accidentes tanto en el ámbito marítimo como en el terrestre (en desarrollo). • Esquema propuesto para el acopio de información obtenida a partir de un simulacro. 10. Esta Tesis es la base para el proyecto e-Flow (nombre de una de las aplicaciones que desarrolladas en esta obra), un proyecto en el que el autor de esta Tesis ha trabajado como Project Manager. En el proyecto participa un consorcio de empresas y la UPM, y tiene como objetivo trasladar a la realidad gran parte de los planteamientos e ideas presentadas en esta Tesis. Este proyecto incluye el desarrollo de la infraestructura física y de servicios que permitirán, entre otras cosas, implementar en infraestructuras complejas una plataforma que posibilita la evacuación dinámica y un control ubicuo de los sistemas de monitorización y actuación implementados. En estos momentos se está finalizando el proyecto, cuyo objetivo final es la implementación de un piloto en un Hospital. También destacamos los siguientes avances a nivel de difusión científico-tecnológico: • Ponencia en el “52 congreso de la Ingeniería Naval en España” presentando un artículo “e-Flow- Sistema integral inteligente de soporte a la evacuación”. En este artículo se trata tanto el proyecto e-Flow del que soy Project Manager, como esta Tesis Doctoral, al ser temas estrechamente vinculados. En 2014 se publicó en dos números de la Revista Ingeniería Naval el artículo presentado a estas jornadas. • Co-autor en el artículo “E-Flow: A communication system for user notification in dynamic evacuation scenarios” presentado en el 7th International Conference on Ubicuous Computing & Ambient Intelligence (UCAMI) celebrado en Costa Rica. Por último, una de las aportaciones más interesantes, es la definición de un gran número de líneas de investigación futuras en base a todos los avances realizados en esta Tesis. ABSTRACT With this Thesis a new approach for understanding evacuation process is considered, taking advantage of the existing and open technologies that will allow this process to be interpreted as a dynamic entity. The methodology involves not only tools that allows on.-time evacuation plans, but also creates a physical insfrastructure that makes possible to feed the system with information on real time so, considering in each moment the real situation as well as the specific emergency development it will be feasible to generate alternative plans that responds to the current emergency situation. In this respect, the system will store all this information and will feedback with solutions that will help the decision making along the evacuation process. The innovative and singular contributions of this Thesis are numerous and rich, summarised as follows: 1.- Complete state-of-art study: a. Detection and analysis of different projects on an international level that, although partially, deal with some aspects developed in this Thesis. b. Thorough study at a international level of the developed software - total or partially done - for the simulation of the human behaviour and evacuation processes analysis. A database has been generated that classifies in detail these applications allowing to perform a full analysis and leading to future evolution of its contents. Within the Thesis work, almost a hundred of developments have been analysed but the purpose is to keep up updating this database due to the broad applications and possibilities that it involves. 2. Development of an important chapter that studies the possibility of using virtual scenarios as mid-term alternative for the use of simulations. This section is divided in two blocks: a. Trials in virtual and real scenarios b. Trials in virutal scenarios (trials performed with several ones). 3. E-Flow net design development: Set of tools developed under Rhinoceros for the evacuation net design based on the elements defined in the Thesis: Nodes, Paths, Relations, Areas 4. E-Flow simulator development: Set of tools that uses Rhinoceros as a 3D simulator of human behaviour. This simulator, of my own design, includes a new and original algorithm of human behaviour that involves aspects that are not found in other simulators. This tool allows to perform groups programmed simulations which behaviour is based on their enviroment analysis and presence of dynamic references. It includes other important innovations as for example: tools for signals analysis, dynamic signal elements, easy obstacle adding etc... More over, a tool that allows the own scenario movement implementation has been created by simulating the own oscillation movement, with the purpose of playing the vessel movement's influences in the individuals' displacements. 5. In an advanced stage of the development, the possibility of generating a video recording of all the simulation was also integrated, then from that moment all tests have been filed (and keep on doing so) in a database that collects all simulation characteristics, failures detected, etc. These stored tests amounts to a total of 81 GB at the moment of finishing the Thesis work. Generation and analysis of paths regarding the evacuation net created with E-Flow design and the simulations performed with E-Flow net Simulator. a. Analysis for the optimisation of the network configuration based in the existing nodes per area. b. Definition of the processes previous to the calculation of the feasible paths c. Paths calculation: i. Analysis of the different algorithms on existance nowadays for the routes optimisation. ii. Development of a new family of algorithms that I have called “Minimum Decision Algorithm (MDA)”, being composed of: 1. MDA basic 2. MDA minimum 3. MDA of not spacial interference 4. MDA of expansion (es de extenderse) o enlargement ( es de crecimiento) 5. MDA of organised expansion for a single origin (of organised enlargement for a single origin) 6. MDA of organised expansion (of organised enlargement) iii. All these algorithms have been implemented in the E-Flow application created in the Thesis dfor the routes analysis and it is the core of the Captain's support system. d. Determination of the alternatives for the evacuation plan. After defining all possible paths, different processes of analysis existing for weighing-based criteria are described, thus to end defining the own development method proposed in this Thesis and that aims to respond in an agreggation of possible routes basis obtained by means of the MDA algorithms what is the routes' combination more suitable for the Plan or Plans in every situation. The methodology created fot the selection of the combinations of routes that determine a complete Plan is baesd in four basic criteria that after applying, offer the best alternatives. In this stage a complex analysis of the progress along time is also included, that adds original and innovative defintions and formulations. e. Originated from the methodology created in this Thesis for the perfoming of the analysy of the progress along time, a new mathematic theorem has been defined, that has been called as "Family of quadrilateral of constant area". 7. Specification of the physiscal infrastructure of the Captain's help system: essential part is this physical infrastructure that will support it. This system will be made of sensors, actuators, apps for mobile devices etc... Within this chapter the different elements and technologies that make up this infrastructure will be studied. 8. Specification for the services infrastructure. 9. Start up of the Blog. " Virtual Environments (http://epcinnova-virtualenvironments.blogspot.com.es/)" in which all tests performed have been published in terms of analysis of the virtual enviroments as alternative to the simulators as well as to the laboratory experiments or simulations, including in most of the cases the possibility that the visitor can perform the simulation within the virtual enviroment. This blog also includes other sections that have been worked along and within this Thesis: - Collection of different virtual scenarios existent. - Schema that gathers information concerning accidents for maritime and terrestrial areas (under development) - Schema proposed for the collecting of information obtained from a simulation. 10. This Thesis is the basis of the E-Flow project (name of one of the applications developed in this work), a project in which the Thesis' author has worked in as Project Manager. In the project takes part a consortium of firms as well as the UPM and the aim is to bring to real life most part of the approaches and ideas contained in this Thesis. This project includes the development of the physical infrastructure as well as the services that will allow, among others, implement in complex infrastrucutres a platform that will make possible a dynamic evacuation and a continuous control of the monitoring and acting systems implemented. At the moment the project is getting to an end which goal is the implementation of a pilot project in a Hospital. We also would like to highlight the following advances concerning the scientific-technology divulgation: • Talk in the " 52th Congress of the Naval Engineering in Spain" with the article "E-Flow . Intelligent system integrated for supporting evacuation". This paper is about project E-Flow which I am Project Manager of, as well as this Thesis for the Doctorate, being both closely related. Two papers published In 2014 in the Naval Engineering Magazine. • Co-author in the article “E-Flow: A communication system for user notification in dynamic evacuation scenarios” [17] introduced in the 7th International Conference on Ubicuous Computing & Ambient Intelligence (UCAMI) held in Costa Rica. Last, but not least, one of the more interesting contributions is the defintion of several lines of research in the future, based on the advances made in this Thesis.

La transformación de las grandes estaciones europeas con la llegada de la Alta Velocidad. El caso de Atocha

La aparición del tren de alta velocidad en Europa en las últimas décadas del siglo XX supuso el resurgir de un medio de transporte en progresivo declive desde la popularización del automóvil y del avión. La decadencia del ferrocarril había supuesto en muchos casos el abandono, o incluso la demolición, de estaciones históricas y el deterioro de su entorno urbano. Como reacción a esa desatención surgió, también en el último cuarto de siglo, una mayor conciencia social preocupada por la conservación del patrimonio construido del ferrocarril. La necesidad de adaptación de las grandes estaciones de ferrocarril para dar servicio al nuevo sistema de transporte, junto con el interés por poner en valor sus construcciones históricas y su céntrico entorno, ha dado como resultado la realización de importantes transformaciones. El objeto de la presente investigación es el estudio de las transformaciones que han sufrido las grandes estaciones europeas del siglo XIX con la llegada del tren de alta velocidad, profundizando de manera especial en el caso más significativo que tenemos en nuestro país: la estación de Atocha. En el ámbito europeo es donde se localizan los ejemplos más relevantes de estaciones que tuvieron gran trascendencia en el siglo XIX y que ahora, con la llegada de la Alta Velocidad, vuelven a recuperar su grandeza. En España, el crecimiento de la Alta Velocidad en los últimos años ha sido extraordinario, hasta situarse como el segundo país del mundo con más kilómetros de líneas de alta velocidad en operación y, en consecuencia, se ha construido un gran número de estaciones adaptadas a este servicio. El caso más notable es el de la estación de Atocha, que desde la llegada del AVE en 1992 hasta el día de hoy, se ha convertido en uno de los complejos ferroviarios más importantes del mundo. El trabajo parte del estudio de otros referentes europeos, como las Gares de París, la estación de St Pancras en Londres y de otras cinco estaciones del centro de Europa –Amsterdam Centraal, Antwerpen Centraal, Köln Hauptbahnhof, Frankfurt (Main) Hauptbahnhof y la Gare de Strasbourg–, para establecer el marco analítico sobre el que se profundiza con la estación de Atocha. El proceso de transformación de la estación de Atocha se ha gestado a través de una serie de proyectos que han ido configurando la estación hasta el momento actual y planteando la previsión de futuro: el proyecto del Plan General de Madrid, el concurso de ideas para el diseño de la estación, la estación de Cercanías, la estación de Alta Velocidad y Largo Recorrido, la ampliación de esta para separar los flujos por niveles, los Estudios Informativos del Nuevo Complejo Ferroviario de la Estación de Atocha y su primera fase de construcción. Estos siete proyectos son objeto de un análisis en tres niveles: análisis cronológico, análisis funcional y análisis formal. La estación de Atocha fue la primera estación histórica europea en sufrir una gran transformación vinculada a la llegada de la Alta Velocidad. Aporta el entendimiento de la estación como un todo y la intermodalidad como sus principales valores, además de la gran mejora urbana que supuso la «operación Atocha», y adolece de ciertas carencias en su desarrollo comercial, vinculadas en parte a la presencia del jardín tropical, y de un pobre espacio en las salas de embarque para los pasajeros de salidas. La estación de Atocha completa su transformación a partir de su renovación funcional, manteniendo la carga simbólica de su historia. De la confrontación del caso de Atocha con otras importantes estaciones europeas resulta la definición de las principales consecuencias de la llegada de la Alta Velocidad a las grandes terminales europeas y la identificación de los elementos clave en su transformación. Las consecuencias principales son: la potenciación de la intermodalidad con otros medios de transporte, el desarrollo comercial no necesariamente destinado a los usuarios de los servicios ferroviarios, y la puesta en valor de la antigua estación y de su entorno urbano. Por su parte, los elementos clave en la transformación de las grandes estaciones tienen que ver directamente con la separación de flujos, el entendimiento de la estación por niveles, la dotación de nuevos accesos laterales y la construcción de una nueva gran cubierta para los nuevos andenes. La preeminencia de unos elementos sobre otros depende del carácter propio de cada estación y de cada país, de la magnitud de la intervención y, también, de la estructura y composición de los equipos encargados del diseño de la nueva estación. En la actualidad, nos encontramos en un momento interesante respecto a las estaciones de Alta Velocidad. Tras el reciente atentado frustrado en el Thalys que viajaba de Ámsterdam a París, se ha acordado establecer controles de identidad y equipajes en todas las estaciones de la red europea de alta velocidad, lo que implicará modificaciones importantes en las grandes estaciones que, probablemente, tomarán el modelo de la estación de Atocha como referencia. ABSTRACT The emergence of the high speed train in Europe in the last few decades of the 20th century represented the resurgence of a means of transport in progressive decline since the popularization of the car and the airplane. The railway decay brought in many cases the abandonment, or even the demolition, of historical stations and the deterioration of its urban environment. In response to that neglect, a greater social awareness towards the preservation of the railway built heritage raised up, also in the last quarter-century. The need for adaptation of the great railway stations to serve the new transport system, along with the interest in enhancing the historical buildings and its central locations, had resulted in important transformations. The subject of current investigation is the study of the transformations that the great 19th century European stations have experienced with the arrival of the high speed rail, deepening in particular in the most significant case we have in Spain: Atocha railway station. At European level is where the most relevant examples of stations which have had a great significance in the 19th century and now, with the arrival of the high speed train, have regain their greatness, are located. In Spain, the growth of the high speed rail over the past few years has been outstanding. Today is the second country in the world with the longest high speed rail network in operation and, therefore, with a great number of new stations adapted to this service. The most remarkable case is Atocha station. Since the arrival of the AVE in 1992, the station has become one of the world's most important railway hub. The research starts with the study of other European reference points, as the Gares of Paris, St Pancras station in London and five other stations of Central Europe –Amsterdam Centraal, Antwerpen Centraal, Köln Hauptbahnhof, Frankfurt (Main) Hauptbahnhof y la Gare de Strasbourg–, to establish the analytical framework that will be deepen with Atocha station. The transformation process of Atocha station has been created through a number of projects that have forged the station to date and have raised the sights in the future: the project of the General Urban Development Plan, the ideas competition for the station design, the Suburban train station, the High Speed and Long Distance station, its enlargement in order to separate passenger flows in different levels, the 'Masterplans' for the new Atocha transport hub and its first phase of construction. These seven projects are under scrutiny at three levels: chronological analysis, functional analysis and formal analysis. Atocha station was the first European historical station to undergo a great transformation tied to the arrival of the high speed rail. It brings the understanding of the station as a whole and the intermodality as its greatest values, besides the great urban improvement of the 'Atocha operation', and suffers from certain shortcomings in its commercial development, partly linked to the presence of the tropical garden, and from a poor space in the departure lounges. Atocha station completes its transformation on the basis of its functional renewal, keeping the symbolic charge of its history. The confrontation of Atocha case with the great European stations results in the definition of the principal consequences of the high speed rail arrival to the great European terminals and the identification of the key elements in its transformation. The principal consequences are: the empowering of the intermodality with other means of transport, of the commercial development, not necessarily intended for railway services users, and the enhancement of the old station and its urban environment. On the other hand, the key elements in the transformation of the great stations are directly related with the separation of passenger flows, the understanding of the station in different levels, the placement of new lateral accesses and the construction of a new deck over the new platforms. The pre-eminence of some elements over the others depends on the particular nature of each station and each country, on the scale of the intervention and also in the structure and composition of the teams in charge of the new station design. Nowadays, this is an interesting time concerning the high speed rail stations. After the recent foiled terrorist attempt in the Thalys train travelling from Amsterdam to Paris, it was agreed to establish passenger and luggage controls in every European high speed rail station. This will mean important changes in these great stations, which probably will take Atocha station's model as a reference.

La casa crecedera. El crecimiento programado de la vivienda con innovación europea y economía de medios latinoamericana

El concepto de casa crecedera, tal y como lo conocemos en la actualidad, se acuñó por primera vez en 1932 en el concurso Das Wachsende Haus organizado por Martin Wagner y Hans Poelzig dentro del marco de la Exposición Internacional Sonne, Luft und Haus für alle, promovida por la Oficina de Turismo de la ciudad de Berlín. En dicho concurso, se definía este tipo de vivienda como aquella célula básica o vivienda semilla que, dependiendo de las necesidades y posibilidades de los habitantes, podía crecer mediante otras estancias, conformando una vivienda completa en sí misma en cada fase de crecimiento. Numerosos arquitectos de primer orden, tales como Walter Gropius, Bruno Taut, Erich Mendelsohn o Hans Scharoun, participaron en este concurso, abriendo una nueva vía de exploración dentro de la vivienda flexible, la del crecimiento programado. A partir de ese momento, en Europa, y subsecuentemente en EEUU y otras regiones desarrolladas, se iniciaron numerosas investigaciones teóricas y prácticas en torno al fenómeno del crecimiento en la vivienda desde un enfoque vinculado a la innovación, tanto espacial como técnica. Por otro lado, aunque dentro del marco de la arquitectura popular de otros países, ya se ensayaban viviendas crecederas desde el siglo XVIII debido a que, por su tamaño, eran más asequibles dentro del mercado. Desde los años treinta, numerosos países en vías de desarrollo tuvieron que lidiar con migraciones masivas del campo a la ciudad, por lo que se construyeron grandes conjuntos habitacionales que, en numerosos casos, estaban conformados por viviendas crecederas. En todos ellos, la aproximación al crecimiento de la vivienda se daba desde una perspectiva diferente a la de los países desarrollados. Se primaba la economía de medios, el uso de sistemas constructivos de bajo costo y, en muchos casos, se fomentaba incluso la autoconstrucción guiada, frente a las construcciones prefabricadas ensambladas por técnicos especializados que se proponían, por ejemplo, en los casos europeos. Para realizar esta investigación, se recopiló información de estas y otras viviendas. A continuación, se identificaron distintas maneras de producir el crecimiento, atendiendo a su posición relativa respecto de la vivienda semilla, a las que se denominó mecanismos de ampliación, utilizados indistintamente sin tener en cuenta la ubicación geográfica de cada casa. La cuestión de porqué se prefiere un mecanismo en lugar de otro en un caso determinado, desencadenó el principal objetivo de esta Tesis: la elaboración de un sistema de análisis y diagnóstico de la vivienda crecedera que, de acuerdo a determinados parámetros, permitiera indicar cuál es la ampliación o sucesión de ampliaciones óptimas para una familia concreta, en una ubicación establecida. Se partió de la idea de que el crecimiento de la vivienda está estrechamente ligado a la evolución de la unidad de convivencia que reside en ella, de manera que la casa se transformó en un hábitat dinámico. Además se atendió a la complejidad y variabilidad del fenómeno, sujeto a numerosos factores socio-económicos difícilmente previsibles en el tiempo, pero fácilmente monitorizables según unos patrones determinados vinculados a la normatividad, el número de habitantes, el ahorro medio, etc. Como consecuencia, para el diseño del sistema de optimización de la vivienda crecedera, se utilizaron patrones evolutivos. Dichos patrones, alejados ya del concepto espacial y morfológico usualmente utilizado en arquitectura por figuras como C. Alexander o J. Habraken, pasaron a entenderse como una secuencia de eventos en el tiempo (espaciales, sociales, económicos, legales, etc.), que describen el proceso de transformación y que son peculiares de cada vivienda. De esta manera, el tiempo adquirió una especial importancia al convertirse en otro material más del proyecto arquitectónico. Fue en la construcción de los patrones donde se identificaron los mencionados mecanismos de ampliación, entendidos también como sistemas de compactación de la ciudad a través de la ocupación tridimensional del espacio. Al estudiar la densidad, mediante los conceptos de holgura y hacinamiento, se aceptó la congestión de las ciudades como un valor positivo. De esta forma, las posibles transformaciones realizadas por los habitantes (previstas desde un inicio) sobre el escenario del habitar (vivienda semilla), se convirtieron también en herramientas de proyecto urbano que responden a condicionantes del lugar y de los habitantes con distintas intensidades de crecimiento, ocupación y densidad. Igualmente, en el proceso de diseño del sistema de optimización, se detectaron las estrategias para la adaptabilidad y transformación de la casa crecedera, es decir, aquella serie de acciones encaminadas a la alteración de la vivienda para facilitar su ampliación, y que engloban desde sistemas constructivos en espera, que facilitan las costuras entre crecimiento y vivienda semilla, hasta sistemas espaciales que permiten que la casa altere su uso transformándose en un hábitat productivo o en un artefacto de renta. Así como los mecanismos de ampliación están asociados a la morfología, se descubrió que su uso es independiente de la localización, y que las estrategias de adaptabilidad de la vivienda se encuentran ligadas a sistemas constructivos o procesos de gestión vinculados a una región concreta. De esta manera, la combinación de los mecanismos con las estrategias caracterizan el proceso de evolución de la vivienda, vinculándola a unos determinados condicionantes sociales, geográficos y por tanto, constructivos. Finalmente, a través de la adecuada combinación de mecanismos de ampliación y estrategias de adaptabilidad en el proyecto de la vivienda con crecimiento programado es posible optimizar su desarrollo en términos económicos, constructivos, sociales y espaciales. Como resultado, esto ayudaría no sólo a mejorar la vida de los habitantes de la vivienda semilla en términos cualitativos y cuantitativos, sino también a compactar las ciudades mediante sistemas incluyentes, ya que las casas crecederas proporcionan una mayor complejidad de usos y diversidad de relaciones sociales. ABSTRACT The growing house concept -as we currently know it- was used for the first time back in 1932 in the competition Das Wachsende Haus organized by Martin Wagner and Hans Poelzig during the International Exhibition Sonne, Luft und Haus für alle, promoted by Berlin's Tourist Office. In that competition this type of housing was defined as a basic cell or a seed house unit, and depending on the needs and capabilities of the residents it could grow by adding rooms and defining itself as a complete house unit during each growing stage. Many world-top class architects such as Walter Gropius, Bruno Taut, Erich Mendelsohn or Hans Scharoun, were part of this competition exploring a new path in the flexible housing field, the scheduled grownth. From that moment and on, in Europe -and subsequently in the USA and other developed areas- many theorical and pragmatical researchs were directed towards the growing house phenomena, coming from an initial approach related to innovation, spacial and technical innovation. Furthermore -inside the traditional architecture frame in other countries, growing houses were already tested in the XVIII century- mainly due to the size were more affordable in the Real State Market. Since the 30's decade many developing countries had to deal with massive migration movements from the countryside to cities, building large housing developments were -in many cases- formed by growing housing units. In all of these developing countries the growing house approach was drawn from a different perspective than in the developed countries. An economy of means was prioritized, the utilization of low cost construction systems and -in many cases- a guided self-construction was prioritized versus the prefabricated constructions set by specialized technics that were proposed -for instance- in the European cases. To proceed with this research, information from these -and other- housing units was gathered. From then and on different ways to perform the growing actions were identified, according to its relative position from the seed house unit, these ways were named as addition or enlargement mechanisms indifferently utilized without adknowledging the geographic location for each house. The question of why one addition mechanism is preferred over another in any given case became the main target of this Thesis; the ellaboration of an analysis and diagnosis system for the growing house -according to certain parameters- would allow to point out which is the addition or addition process more efficient for a certain family in a particular location. As a starting point the grownth of the housing unit is directly linked to the evolution of the family unit that lives on it, so the house becomes a dynamic habitat. The complexity and the variability of the phenomena was taken into consideration related to a great number of socio-economic factors hardly able to be foreseen ahead on time but easy to be monitored according to certain patterns linked to regulation, population, average savings, etc As a consequence, to design the optimization system for the growing house, evolutionary patterns were utilized. Those patterns far away from the spatial and morphologic concept normally utilized in Architecture by characters like C. Alexander or J. Habraken, started to be understood like a sequence of events on time (spatial events, social events, economic events, legal events, etc) that describes the transformation process and that are particular for each housing unit. Therefore time became something important as another ingredient in the Architectural Project. The before mentioned addition or enlargement mechanisms were identified while building the patterns; these mechanisms were also understood as city's system of compactation through the tridimendional ocupation of space. Studying density, thorough the concepts of comfort and overcrowding, traffic congestion in the city was accepted as a positive value. This way, the possible transformations made by the residents (planned from the begining) about the residencial scenary (seed house), also became tools of the urban project that are a response to site's distinctive features and to the residents with different grownth intensities, activities and density Likewise, during the process of designing the optimization system, strategies for adaptations and transformation of the growing house were detected, in other words, the serial chain of actions directed to modify the house easing its enlargement or addition, and that comprehends from constructive systems on hold -that smooths the costures between grownth and housing seed- to spatial systems that allows that the house modify its utilization, becoming a productive habitat or a rental asset. Because the enlargement mechanisms are linked to the morphology, it was discovered that the use it's not related to the location, and that the adaptation strategies of the houses are linked to constructive systems or management processes linked to a particular area. This way the combination of mechanisms and strategies characterizes the process of housing evolution, linking it to certain social and geographic peculiarities and therefore constructives. At last, through the certain combination of enlargement mechanisms and adaptability strategies in the housing with scheduled grownth project is possible to optimize its development in economic, constructive, social and spatial terms. As a result, this would help not only to improve the life of the seed house residents in qualitative and quantitative terms but also to compact the cities through inclusive systems, given that the growing houses provide a larger complexity of uses and social relations.

Characterization of mice deficient in aromatase (ArKO) because of targeted disruption of the cyp19 gene

The formation of estrogens from C19 steroids is catalyzed by aromatase cytochrome P450 (P450arom), the product of the cyp19 gene. The actions of estrogen include dimorphic anatomical, functional, and behavioral effects on the development of both males and females, considerations that prompted us to examine the consequences of deficiency of aromatase activity in mice. Mice lacking a functional aromatase enzyme (ArKO) were generated by targeted disruption of the cyp19 gene. Male and female ArKO mice were born with the expected Mendelian frequency from F1 parents and grew to adulthood. Female ArKO mice at 9 weeks of age displayed underdeveloped external genitalia and uteri. Ovaries contained numerous follicles with abundant granulosa cells and evidence of antrum formation that appeared arrested before ovulation. No corpora lutea were present. Additionally the stroma were hyperplastic with structures that appeared to be atretic follicles. Development of the mammary glands approximated that of a prepubertal female. Examination of male ArKO mice of the same age revealed essentially normal internal anatomy but with enlargement of the male accessory sex glands because of increased content of secreted material. The testes appeared normal. Male ArKO mice are capable of breeding and produce litters of approximately average size. Whereas serum estradiol levels were at the limit of detection, testosterone levels were elevated, as were the levels of follicle-stimulating hormone and luteinizing hormone. The phenotype of these animals differs markedly from that of the previously reported ERKO mice, in which the estrogen receptor α is deleted by targeted disruption.

Relax promotes ectopic neuronal differentiation in Xenopus embryos

We previously isolated a novel rat cDNA encoding a basic helix–loop–helix transcription factor named Relax, whose expression in the developing central nervous system is strictly limited to discrete domains containing precursor cells. The timing of Relax expression coincides with neuronal differentiation. To investigate the involvement of Relax in neurogenesis we tested whether Relax activated neural genes in the ectoderm by injecting Relax RNA into Xenopus embryos. We demonstrate that ectopic Relax expression induces a persistent enlargement of the neural plate and converts presumptive epidermal cells into neurons. This indicates that Relax, when overexpressed in Xenopus embryos, has a neuronal fate-determination function. Analyses both of Relax overexpression in the frog and of the distribution of Relax in the rat neural tube strongly suggest that Relax is a neuronal fate-determination gene.

A tetracycline derivative, minocycline, reduces inflammation and protects against focal cerebral ischemia with a wide therapeutic window

The only treatment of patients with acute ischemic stroke is thrombolytic therapy, which benefits only a fraction of stroke patients. Both human and experimental studies indicate that ischemic stroke involves secondary inflammation that significantly contributes to the outcome after ischemic insult. Minocycline is a semisynthetic second-generation tetracycline that exerts antiinflammatory effects that are completely separate from its antimicrobial action. Because tetracycline treatment is clinically well tolerated, we investigated whether minocycline protects against focal brain ischemia with a wide therapeutic window. Using a rat model of transient middle cerebral artery occlusion, we show that daily treatment with minocycline reduces cortical infarction volume by 76 ± 22% when the treatment is started 12 h before ischemia and by 63 ± 35% when started even 4 h after the onset of ischemia. The treatment inhibits morphological activation of microglia in the area adjacent to the infarction, inhibits induction of IL-1β-converting enzyme, and reduces cyclooxygenase-2 expression and prostaglandin E2 production. Minocycline had no effect on astrogliosis or spreading depression, a wave of ionic transients thought to contribute to enlargement of cortical infarction. Treatment with minocycline may act directly on brain cells, because cultured primary neurons were also salvaged from glutamate toxicity. Minocycline may represent a prototype of an antiinflammatory compound that provides protection against ischemic stroke and has a clinically relevant therapeutic window.

Actin Filaments and Microtubules are Involved in Different Membrane Traffic Pathways That Transport Sphingolipids to the Apical Surface of Polarized HepG2 Cells

In polarized HepG2 hepatoma cells, sphingolipids are transported to the apical, bile canalicular membrane by two different transport routes, as revealed with fluorescently tagged sphingolipid analogs. One route involves direct, transcytosis-independent transport of Golgi-derived glucosylceramide and sphingomyelin, whereas the other involves basolateral to apical transcytosis of both sphingolipids. We show that these distinct routes display a different sensitivity toward nocodazole and cytochalasin D, implying a specific transport dependence on either microtubules or actin filaments, respectively. Thus, nocodazole strongly inhibited the direct route, whereas sphingolipid transport by transcytosis was hardly affected. Moreover, nocodazole blocked “hyperpolarization,” i.e., the enlargement of the apical membrane surface, which is induced by treating cells with dibutyryl-cAMP. By contrast, the transcytotic route but not the direct route was inhibited by cytochalasin D. The actin-dependent step during transcytotic lipid transport probably occurs at an early endocytic event at the basolateral plasma membrane, because total lipid uptake and fluid phase endocytosis of horseradish peroxidase from this membrane were inhibited by cytochalasin D as well. In summary, the results show that the two sphingolipid transport pathways to the apical membrane must have a different requirement for cytoskeletal elements.

Phosphatidylinositol 3-kinase signaling mediates angiogenesis and expression of vascular endothelial growth factor in endothelial cells

Phosphatidylinositol 3-kinase (PI 3-kinase) is a signaling molecule that controls numerous cellular properties and activities. The oncogene v-p3k is a homolog of the gene coding for the catalytic subunit of PI 3-kinase, p110α. P3k induces transformation of cells in culture, formation of hemangiosarcomas in young chickens, and myogenic differentiation in myoblasts. Here, we describe a role of PI 3-kinase in angiogenesis. Overexpression of the v-P3k protein or of cellular PI 3-kinase equipped with a myristylation signal, Myr-P3k, can induce angiogenesis in the chorioallantoic membrane (CAM) of the chicken embryo. This process is characterized by extensive sprouting of new blood vessels and enlargement of preexisting vessels. Overexpression of the myristylated form of the PI 3-kinase target Akt, Myr-Akt, also induces angiogenesis. Overexpression of the tumor suppressor PTEN or of dominant-negative constructs of PI 3-kinase inhibits angiogenesis in the yolk sac of chicken embryos, suggesting that PI 3-kinase and Akt signaling is required for normal embryonal angiogenesis. The levels of mRNA for vascular endothelial growth factor (VEGF) are elevated in cells expressing activated PI 3-kinase or Myr-Akt. VEGF mRNA levels are also increased by insulin treatment through the PI 3-kinase-dependent pathway. VEGF mRNA levels are decreased in cells treated with the PI 3-kinase inhibitor LY294002 and restored by overexpression of v-P3k or Myr-Akt. Overexpression of VEGF by the RCAS vector induces angiogenesis in chicken embryos. These results suggest that PI 3-kinase plays an important role in angiogenesis and regulates VEGF expression.

Regulation of dorsal fate in the neuraxis by Wnt-1 and Wnt-3a

Members of the Wnt family of signaling molecules are expressed differentially along the dorsal–ventral axis of the developing neural tube. Thus we asked whether Wnt factors are involved in patterning of the nervous system along this axis. We show that Wnt-1 and Wnt-3a, both of which are expressed in the dorsal portion of the neural tube, could synergize with the neural inducers noggin and chordin in Xenopus animal explants to generate the most dorsal neural structure, the neural crest, as determined by the expression of Krox-20, AP-2, and slug. Overexpression of Wnt-1 or Wnt-3a in the neuroectoderm of whole embryos led to a dramatic increase of slug and Krox-20-expressing cells, but the hindbrain expression of Krox-20 remained unaffected. Enlargement in the neural crest population could occur even when cell proliferation was inhibited. Wnt-5A and Wnt-8, neither of which is expressed in the dorsal neuroectoderm, failed to induce neural crest markers. Overexpression of glycogen synthase kinase 3, known to antagonize Wnt signaling, blocked the neural-crest-inducing activity of Wnt-3a in animal explants and inhibited neural crest formation in whole embryos. We suggest that Wnt-1 and Wnt-3a have a role in patterning the neural tube along its dorsoventral axis and function in the differentiation of the neural crest.

A FYVE-finger-containing protein, Rabip4, is a Rab4 effector involved in early endosomal traffic

The small GTPase Rab4 is implicated in endocytosis in all cell types, but also plays a specific role in some regulated processes. To better understand the role of Rab4 in regulation of vesicular trafficking, we searched for an effector(s) that specifically recognizes its GTP-bound form. We cloned a ubiquitous 69-kDa protein, Rabip4, that behaves as a Rab4 effector in the yeast two-hybrid system and in the mammalian cell. Rabip4 contains two coiled-coil domains and a FYVE-finger domain. When expressed in CHO cells, Rabip4 is present in early endosomes, because it is colocated with endogenous Early Endosome Antigen 1, although it is absent from Rab11-positive recycling endosomes and Rab-7 positive late endosomes. The coexpression of Rabip4 with active Rab4, but not with inactive Rab4, leads to an enlargement of early endosomes. It strongly increases the degree of colocalization of markers of sorting (Rab5) and recycling (Rab11) endosomes with Rab4. Furthermore, the expression of Rabip4 leads to the intracellular retention of a recycling molecule, the glucose transporter Glut 1. We propose that Rabip4, an effector of Rab4, controls early endosomal traffic possibly by activating a backward transport step from recycling to sorting endosomes.