Efecto de la infección por virus persistentes y no persistentes sobre el comportamiento alimenticio, la preferencia, tasa de aterrizaje y asentamiento y la eficacia biológica de Aphis gossypii Glover

En el complejo de plagas que atacan a los principales cultivos hortícolas protegidos, destacan principalmente los Hemípteros, y dentro de estos los pulgones, dada su importancia como vectores de virus que provocan considerables daños y pérdidas económicas. Debido a que la dispersión de la mayoría de los virus de plantas puede ser eficaz con densidades bajas de vectores y su control es muy complicado al no existir métodos curativos para su control, es necesario generar nuevos conocimientos sobre las interacciones virus-vector con el fin de desarrollar nuevas y eficaces estrategias de control. Por ello, el objetivo general de esta Tesis ha sido conocer el efecto de la infección viral (directo-mediado por la presencia del virus en el vector- e indirecto-mediado por las alteraciones físico-químicas que se originan en la planta como consecuencia de la infección viral-) sobre el comportamiento y eficacia biológica del vector Aphis gossypii Glover y sus posibles repercusiones en la epidemiología de virosis de transmisión no persistente (Cucumber mosaic virus, CMV, Cucumovirus) y persistente (Cucurbit aphid-borne yellows virus, CABYV, Polerovirus). El primer objetivo de esta Tesis Doctoral, se centró en el estudio del efecto indirecto del virus de transmisión no persistente CMV sobre el comportamiento alimenticio y la preferencia del pulgón A. gossypii en el cultivo de pepino. Los ensayos de despegue y aterrizaje mostraron que los pulgones que fueron liberados en las plantas de pepino infectadas con CMV tuvieron una mayor propensión en migrar hacia las plantas no infectadas (60, 120 y 180 minutos después de la liberación) que aquellos que fueron sometidos al tratamiento contrario (planta no infectada hacia planta infectada con CMV). El estudio de preferencia y asentamiento mostró que el vector A. gossypii prefiere asentarse en plantas infectadas con CMV en una etapa temprana de evaluación (30 minutos después de la liberación). Sin embargo, este comportamiento se revirtió en una etapa posterior (4 y 48 horas después de la liberación), donde los pulgones se asentaron más en las plantas no infectadas. A través de la técnica de Gráficos de Penetración Eléctrica (EPG) se observó un efecto indirecto del virus CMV, revelado por un cambio brusco en el comportamiento de prueba del pulgón a lo largo del tiempo, cuando éstos fueron expuestos a las plantas infectadas con CMV. Los primeros 15 minutos de registro EPG mostraron que los pulgones hicieron un número mayor de punciones intracelulares (potencial drops - pds) y pruebas en las plantas infectadas con CMV que en las plantas no infectadas. Por otra parte, la duración de la primera prueba fue más corta y la duración total de las pds por insecto fue mucho más larga en las plantas infectadas con CMV. Se observaron diferencias significativas en el tiempo transcurrido desde el final de la última pd hasta el final de la prueba, siendo ese tiempo más corto para los pulgones que estaban alimentándose en plantas infectadas con CMV. En la segunda hora de registro los pulgones rechazaron las plantas infectadas con CMV como fuente de alimento, permaneciendo menos tiempo en las fases de prueba en floema (fase de salivación – E1 y fase de ingestión del floema – E2). El comportamiento alimenticio observado sobre las plantas infectadas con CMV favorece la adquisición y posterior transmisión de los virus de transmisión no persistente, los cuales son adquiridos e inoculados durante la realización de pruebas intracelulares en las primeras pruebas de corta duración. En el segundo objetivo de la Tesis se evaluó el efecto directo e indirecto del virus de transmisión persistente CABYV en el comportamiento alimenticio y preferencia del pulgón A. gossypii en cultivo de pepino, especie susceptible al virus, y algodón, especie inmune al virus. No se observó un efecto directo del virus relevante en el comportamiento alimenticio del vector, ya que los resultados obtenidos a nivel floemático en plantas de pepino no se observaron en plantas de algodón, inmune al virus CABYV. Esto sugiere que los resultados obtenidos en pepino, pueden deberse a un “posible efecto indirecto” originado por la infección de las plantas susceptibles al virus durante la realización del ensayo, lo que indirectamente puede modificar el comportamiento del pulgón durante la fase de evaluación. Sin embargo, el virus CABYV modificó indirectamente el comportamiento alimenticio de su vector a través de cambios en la planta infectada. Los pulgones tardaron menos tiempo en llegar al floema, realizaron un mayor número de pruebas floemáticas y permanecieron durante más tiempo en actividades floemáticas en plantas infectadas con CABYV. El comportamiento observado sobre las plantas infectadas con CABYV favorece la adquisición de virus persistentes, los cuales son adquiridos durante la alimentación sostenida en floema. El estudio de preferencia y asentamiento de A. gossypii mostró que los pulgones virulíferos prefieren asentarse en plantas no infectadas a corto y largo plazo de evaluación (2, 4 y 48 horas después de la liberación). Los ensayos de despegue y aterrizaje mostraron que los pulgones virulíferos que fueron liberados en las plantas de pepino infectadas con CABYV tuvieron una mayor propensión en migrar hacia las plantas no infectadas (3, 6, 24 y 48 horas después de la liberación) que aquellos que fueron sometidos al tratamiento contrario (planta no infectada hacia planta infectada con CABYV). Sin embargo, los pulgones no virulíferos no mostraron preferencia por plantas de pepino no infectadas o infectadas con CABYV en ninguno de los ensayos (preferencia o despegue) o periodos evaluados (corto y largo plazo). Los resultados indican que el virus CABYV es capaz de modificar indirectamente el comportamiento alimenticio de su vector a través de cambios en la planta infectada, favoreciendo su adquisición por su principal vector, A. gossypii. Una vez que los pulgones tienen capacidad de transmitir el virus (virulíferos) se produce un cambio en su comportamiento prefiriendo asentarse sobre plantas no infectadas optimizándose así la dispersión viral. El tercer objetivo de la Tesis, fue evaluar los efectos directos e indirectos del virus CABYV así como los efectos indirectos del virus CMV en la eficacia biológica del vector A. gossypii. Los resultados obtenidos en los ensayos realizados con el virus persistente CABYV indican que el virus parece no modificar directamente ni indirectamente la eficacia biológica del vector en plantas de pepino o algodón, no observándose diferencias estadísticas en ninguno de los parámetros poblacionales evaluados (tiempo de desarrollo, tasa intrínseca de crecimiento, tiempo generacional medio, tasa media de crecimiento relativo y ninfas totales). En cuanto a los ensayos realizados con el virus no persistente, CMV, los resultados muestran un efecto indirecto del virus sobre la biología del vector. Así resultó que tanto la tasa intrínseca de crecimiento natural (rm) como la tasa media de crecimiento relativo (RGR) fueron más altas para pulgones crecidos sobre plantas infectadas con CMV que sobre plantas no infectadas, favoreciendo la reproducción y crecimiento poblacional del vector sobre plantas infectadas con CMV. Los resultados obtenidos en la presente Tesis, ofrecen un ejemplo de como los virus de plantas pueden manipular directa e indirectamente a su vector, maximizando así su dispersión entre las plantas. Esos nuevos conocimientos generados tienen implicaciones importantes en la transmisión, dispersión y en la epidemiología de los virus y deben ser considerados para diseñar o ajustar los modelos de simulación existentes y patrones de dispersión que describen las epidemias de estos virus. ABSTRACT The main objective of this Thesis has been to understand the effect of the viral infection (direct-mediated by the presence of the virus in the vector and indirect mediated by the chemical and physical changes originated in the plant as a consequence of the viral infection) on the behaviour and biological efficacy of the vector Aphis gossypii Glover and its consequences in the epidemiology of two viral diseases, one with non-persistent transmission (Cucumber mosaic virus, CMV, Cucumovirus) and another with persistent transmission (Cucurbit aphid-borne yellows virus, CABYV, Polerovirus). The first objective of this Thesis was the study of the indirect effect of the nonpersistent virus CMV on the feeding behaviour and preference of the aphid A. gossypii in cucumber plants. The results of the alighting and settling behaviour studies showed that aphids exhibited no preference to migrate from CMV-infected to mock-inoculated plants at short time intervals (1, 10 and 30 min after release), but showed a clear shift in preference to migrate from CMV-infected to mock-inoculated plants 60 min after release. Our free-choice preference assays showed that A. gossypii alates preferred CMV-infected over mockinoculated plants at an early stage (30 min), but this behaviour was reverted at a later stage and aphids preferred to settle and reproduce on mock-inoculated plants. The electrical penetration graph (EPG) technique revealed a sharp change in aphid probing behaviour over time when exposed to CMV-infected plants. At the beginning (first 15 min) aphid vectors dramatically increased the number of short superficial probes and intracellular punctures when exposed to CMV-infected plants. At a later stage (second hour of recording) aphids diminished their feeding on CMV-infected plants as indicated by much less time spent in phloem salivation and ingestion (E1 and E2). This particular probing behaviour including an early increase in the number of short superficial probes and intracellular punctures followed by a phloem feeding deterrence is known to enhance the transmission efficiency of viruses transmitted in a NP manner. We conclude that CMV induces specific changes in a plant host that modify the alighting, settling and probing behaviour of its main vector A. gossypii, leading to optimum transmission and spread of the virus. The second objective of this work was to evaluate the effects that the persistently aphid transmitted Cucurbit aphid-borne yellows virus (CABYV) can induce directly and indirectly on the alighting, settling and probing behaviour activities of the cotton aphid A. gossypii. Only minor direct changes on aphid feeding behaviour was observed due to CABYV when viruliferous aphids fed on mock-inoculated plants. However, the feeding behaviour of non-viruliferous aphids was very different on CABYV-infected than on mockinoculated plants. Non-viruliferous aphids spent longer time feeding from the phloem when plants were infected by CABYV than on mock-inoculated plants, suggesting that CABYV indirectly manipulates aphid feeding behaviour through its shared host plant in order to favour viral acquisition. The vector alighting and settling preference was compared between nonviruliferous and viruliferous aphids. Viruliferous aphids showed a clear preference for mockinoculated over CABYV-infected plants at short and long time, while such behaviour was not observed for non-viruliferous aphids. Overall, our results indicate that CABYV induces changes in its host plant that modifies aphid feeding behaviour in a way that virus acquisition from infected plants is enhanced. Once the aphids become viruliferous they prefer to settle on healthy plants, leading to optimize the transmission and spread of the virus. The third objective was to evaluate the direct and indirect effects of CABYV and indirect effects of the CMV on the A. gossypii fitness. Obtained results for the persistent virus CABYV showed that the virus did not modify the vector fitness in cucumber or cotton plants. None of the evaluated variables was statistically significant (development time (d), intrinsic growth rate (rm), mean relative growth rate (RGR) and total number of nymphs). On the other hand, data obtained for the non-persistent virus (CMV) showed an indirect effect of the virus on the vector fitness. Thus, the rm and RGR were higher for aphids grown on CMV-infected plants compared to aphids grown on mock-inoculated plants. Overall, the obtained results are clear examples of how plant viruses could manipulate directly and indirectly vector behaviour to optimize its own dispersion. These results are important for a better understanding of transmission, dispersion and epidemiology of plant viruses transmitted by vectors. This information could be also considered to design or adjust simulation models and dispersion patterns that describe plant virus epidemics.

Elementos peatonales de las ciudades medias españolas : tipos, orígenes, relaciones y articulaciones

A través de esta tesis se busca aportar un análisis de los elementos peatonales que cubren las necesidades y las infraestructuras de apoyo al peatón, siendo un campo poco desarrollado en nuestro país a diferencia de otros como Dinamarca, Reino Unido, Alemania y Holanda, donde las actuaciones peatonales ejecutadas son tradicionales y muy exitosas. Se trata por tanto de un tema poco estudiado pero que adquiere singular importancia en estos años de búsqueda de nuevos modelos urbanos que eviten el derroche de recursos, y avancen hacia formas de movilidad más adecuadas a un mundo limitado y vulnerable. La investigación tiene un carácter eminentemente descriptivo cuyo principal valor es documentar y analizar sistemáticamente una realidad poco conocida, aportándose datos para entender el origen y desarrollo de las actuaciones peatonales. El propósito es ayudar a cimentar este campo en nuestro país, definiendo con precisión el estado de la cuestión geográficamente y dejando para un futuro una cartografía temática completa. El objeto general es estudiar y comprender los elementos peatonales de un número limitado de ciudades medias españolas, marcándose como objetivos específicos los siguientes: - Identificar y describir los principales elementos que conforman la red peatonal de las ciudades. ‐ Estudiar las tipologías espaciales dominantes, sus características y orígenes, pudiendo distinguir de un lado, las desarrolladas en la ciudad consolidada, como las actuaciones en los cascos antiguos; y por otro, las ejecutadas en la ciudad nueva a través de fórmulas arquitectónicas generalmente residenciales con espacios peatonales adjuntos. ‐ Analizar los referidos elementos para comprobar si han conformado redes de uso peatonal, si se han constituido articulando la ciudad como en el caso de los ejes lineales en las ciudades con río o mar, o si han influenciado el carácter del entorno en el que se han realizado, centrando el interés en los cascos históricos peatonalizados. ‐ Comprobar si hay actuaciones complementarias a los procesos de urbanización y mejora urbanística, así como la aprobación de ayudas y subvenciones para afrontar las mismas. ‐ Finalmente, describir y evaluar los procesos y procedimientos mediante los que se produce la peatonalización de determinadas áreas urbanas, los orígenes, los instrumentos y fechas de su puesta en marcha, aportando una cronología peatonal. La metodología comienza con la acotación de 22 ciudades españolas, capitales de provincia con una población entre 100.000 y 500.000 habitantes según los datos del INE del año 2009. La distribución es equilibrada con la selección de ciudades de costa, de interior, con río/ría, con carácter patrimonial, ciudades en islas, etc. El análisis de los elementos peatonales de estas urbes se realiza gracias a la gran resolución de las bases cartográficas públicas y privadas que hay en la actualidad en internet, especialmente con el uso de las herramientas de Google Earth y Google Maps en su versión “street view”, softwares que permiten un paseo virtual de cada ciudad detallándose con gran precisión las actuaciones peatonales. Las ciudades elegidas son: ‐ 11 ciudades costeras: Alicante, Almería, Bilbao, Cádiz, Castellón de la Plana, Huelva, Las Palmas de Gran Canaria, Palma de Mallorca, Santander, San Sebastián y Tarragona. ‐ 11 ciudades de interior: Albacete, Burgos, Córdoba, León, Lérida, Logroño, Oviedo, Murcia, Pamplona, Valladolid y Vitoria. Finalmente, por tener los porcentajes mas elevados de las áreas peatonales de sus casco histórico, se eligen 3 casos de ciudades representativas para un desarrollo peatonal en profundidad, seleccionándose Burgos, San Sebastián y Pamplona. Otros aspectos metodológicos interesantes a destacar, es la extensa documentación adquirida a través de las fuentes de Internet, sobre todo a la hora de desarrollar el último capítulo “Procesos de peatonalización de las ciudades españolas” centrado en las 3 ciudades elegidas. Se consulta las hemerotecas de periódicos locales, los boletines oficiales, las actas de los plenos de los Ayuntamientos y los documentos de planificación (Planes Generales, Planes Especiales, Planes y Proyectos Municipales, Planes de Movilidad), adquiriéndose una precisa información en cuanto a la cronología de las fechas y extensión del trabajo. Se complementa la metodología con las entrevistas a actores claves en el proceso peatonal, como el personal técnico de los Ayuntamiento y los técnicos redactores de los proyectos; y con la colaboración directa de los técnicos de los consistorios de las 3 ciudades seleccionadas, aportando todo tipo de información demandada. En la tesis, se introduce los elementos peatonales mediante el estudio de los espacios peatonales y los procesos de peatonalización existentes, a través del análisis de la literatura consultada aportándose información valiosa en cuanto a la definición de los modelos peatonales, exponiéndose una breve historia de las peatonalizaciones. Destacan las publicaciones de los expertos europeos en la materia como los autores Carmen Hass-Klau, Rolf Monheim, Rob Krier y Collin Buchanan. En España, las actuaciones peatonales fueron tardías en el tiempo en comparación con el resto de Europa, con casi 40 años de retraso. Las referencias bibliográficas españolas en esta materia son escasas siendo las mas significativas las realizadas por Alfonso Sanz y por el profesor Julio Pozueta destacando una de sus últimas publicaciones “La ciudad paseable” describiéndose los elementos peatonales existentes en las ciudades, sirviendo como modelo y ejemplo para esta investigación. En base a lo anterior, se definen los elementos peatonales como aquellos espacios públicos libres de edificación que, por sus características y diseño, garantizan un confortable uso estancial, de tránsito peatonal y están expresamente reservados para ello. En la tesis se detectan y se especifican los siguientes elementos peatonales: las calles y plazas peatonales, las aceras y bulevares de una anchura superior a 8 metros, los paseos peatonales, los elementos de paso (tradicionales y mecánicos), espacios peatonales ligados a nuevas áreas residenciales, los soportales y pasajes peatonales con una anchura mínima de 4 metros, recintos feriales, espacios libres y zonas verdes y parques. Para cada una de las 22 ciudades seleccionadas se realiza un exhaustivo inventario de los anteriores elementos a través de fichas, aportando un gran número de proyectos individuales ejecutados en cada urbe. Esta información se recoge gráficamente en 2 fotoplanos, conformándose como herramientas fundamentales para esta investigación. El Fotoplano 1 se hace sobre la base de la ciudad del Google Earth diferenciándose las áreas peatonales y las zonas verdes. En el Fotoplano 2, y con el soporte anterior, se distinguen las tipologías persistentes y dominantes como son los grandes ejes peatonales, las redes peatonales locales y los elementos peatonales singulares. Finalmente, a partir de los 2 fotoplanos, se realiza la denominada “Huella peatonal” que destaca por su atractivo y pregnancia, algo de lo que el campo de lo peatonal es muy útil. A través de la huella se obtiene una rápida información al tener el plano dos colores, color naranja para lo peatonal (formado por las áreas peatonales y zonas verdes) y color negro para el resto el resto no peatonal. Toda esta documentación gráfica y descriptiva de los elementos peatonales de cada ciudad, queda ampliamente recogida en el Anexo de la tesis. La “Huella peatonal” se considera como el plano básico en la investigación obteniendo una percepción rápida, clara y visual del espacio peatonal de cada ciudad. Se arroja información morfológica con respecto a la distribución espacial, articulación, equilibrio, modulación, cohesión, concentración peatonal, etc. de cada una de las ciudades seleccionadas. A través de la huella se resuelve muchos de las cuestiones enmarcadas como objetivos de la tesis, comprobándose de forma general, que los elementos peatonales dentro de la trama urbana están desarticulados ya que no hay una estructura que module la ciudad como un conjunto cohesionado. Se localizan pequeñas redes conectadas entre sí, que de forma heterogénea, están dispersas por la ciudad. Se verifica que los elementos peatonales con mayor concentración y presencia en las ciudades analizadas son los ejes peatonales, los cascos históricos y los elementos peatonales en las nuevas áreas residenciales. En la investigación, además de estos 3 elementos mencionados, se analiza con mayor profundidad otros tipos que destacan por su persistencia y repetición en la “Huella peatonal” de cada una de las 22 ciudades, como son las plazas peatonales, los soportales y los elementos singulares peatonales: espacios tradicionales (recintos feriales, pasajes, puentes históricos), nuevos elementos peatonales (pasarelas peatonales, elementos mecánicos de paso), áreas peatonales entorno a edificios culturales y otros casos como los consistentes en la recuperación y transformación de edificaciones y construcciones urbanas de diferente uso y tipología, en nuevas áreas peatonales. Se aporta luz sobre su definición, localización, cronología, proporción, presencia y génesis, facilitando resultados de cada uno de ellos. De forma paralela, se obtiene información de índole cuantitativa a través de las mediciones de la huella, obteniéndose datos y porcentajes de extensión de la ciudad, de las zonas peatonales y de las zonas verdes, de los cascos históricos y de sus superficies peatonales, de los metros lineales de dichos cascos históricos y de sus calles peatonales. Se proporcionan ratios e índices que se consideran fundamentales para una mejor comprensión peatonal de cada urbe. Se establece una clasificación de las ciudades en cada uno de los ámbitos descritos, destacando las ciudades de Pamplona, San Sebastián y Burgos por arrojar las cifras peatonales más positivas encabezando la mayoría de las tablas clasificatorias. La peor ciudad valorada es Alicante, seguida según criterio, de Almería, Palma de Mallorca y Las Palmas de GC. En el capítulo final de la investigación, se eligen las ciudades de Burgos, San Sebastián y Pamplona como casos de estudio representativos. El objetivo es obtener un conocimiento más preciso en cuanto a su origen, cronología y procesos de las peatonalizaciones. Cada ciudad se compone de un cuadro cronológicos por etapas, desarrollándose de forma exhaustiva cada una de ellas, enunciadas normalmente, como instrumentos de planeamiento o proyectos de urbanización, acompañadas de los planos de las actuaciones peatonales. Al final de cada etapa se aportan datos de las cifras de las peatonalizaciones que se van realizando, con un cuadro numérico de localizaciones, superficie peatonal, superficie reurbanizada, longitud peatonal y el grado de peatonalización del casco histórico, además de los datos acumulados por etapa. De las conclusiones cruzadas de estas 3 ciudades, destaca la comprobación de la tendencia de ir peatonalizando los centros antiguos llegando incluso al 100% de calles peatonales en el caso de San Sebastián. No obstante, es difícil confirmar si el objetivo es una peatonalización global de los cascos, ya que son muchas las variables que afectan e influyen en el proceso peatonal de cada ciudad. Se coteja adicionalmente cómo se extiende la cultura peatonal desde el interior al exterior, mas allá de los cascos históricos hacia los ensanches próximos. En cuanto al origen de las peatonalizaciones, se concluye que no hay una concepción inicial y teórica marcada, a partir de la cual se va desarrollando la trama peatonal de cada ciudad. Se puede afirmar que los procesos peatonales ejecutados son variados y de diversa índole, sin poder precisar un hecho como causa principal. Según el momento y las circunstancias, las peatonalizaciones responden a distintas iniciativas promovidas por los comerciantes, por las administraciones locales o por los técnicos de planeamiento, con sus expertas aportaciones en los planes. Estos tres agentes actúan como impulsores de proyectos individuales peatonales que se van solapando en el tiempo. Como punto final de la tesis, se propone una serie de nuevas líneas de investigación que pueden servir como estudio adicional y complementario, respondiendo a aspectos tan relevantes como cuestiones de índole económica, social y de movilidad, fuertemente ligadas a las peatonalizaciones, tal y como se demuestra en la literatura consultada. ABSTRACT This thesis aims to analyse the pedestrian elements dealing with the needs and infrastructures that support pedestrians, as this is a field that is little developed in our country, in comparison to other regions such as Denmark, the United Kingdom, Germany and the Netherlands, where the pedestrian actions implemented are traditional and highly successful. Therefore, even though little research has been done about this topic, this field is increasingly important throughout these years of search for urban models intended to prevent the waste of resources and to develop new mobility ways, more adapted to a limited, vulnerable world. This research is essentially descriptive and it mainly aims to document and systematically analysed a scarcely known reality by providing data in order to understand the origin and development of pedestrian actions. This thesis intends to lay the foundations of this field in Spain, accurately defining the state of the art from a geographical point of view and preparing a full thematic map that may be used in the future. The overall aim is to study and understand the pedestrian elements of a limited number of Spanish medium-size cities, establishing the following specific goals: - To identify and describe the main elements comprising cities' pedestrian networks. - To study of special predominant typologies, their features and origins, with the possibility to, on the one hand, make a difference between the typologies developed in well-established cities, such as the actions in historic quarters, and, on the other hand, those implemented in new cities through generally residential architectural formulae with adjoining pedestrian areas. - To analyse the said elements in order to check whether they have resulted in pedestrian-use networks, whether they have been established by organising the city, such as the linear axes in coastal and riverside cities, or whether they have had an impact on the character of the areas where these elements have been implemented, focusing on pedestrian historic quarters. - Check whether there are actions supplementary to the urban development and urban improvement processes, as well as the approval of financial support and subventions to deal with these actions. - Finally, to describe and assess the processes and procedures by which the pedestrianisation of certain urban areas is carried out, the origins, the instruments and the date of their implementation, providing a pedestrian timeline. The methodology starts by defining 22 Spanish cities, province capitals with a population ranging from 100,000 to 500,000 inhabitants, according to the data recorded by the Spanish Statistics Institute (INE) in 2009. The distribution of coastal, riverside and interior cities, as well as of patrimonial cities and cities in islands, etc. is well balanced. The analysis of the pedestrian elements of these cities is made with the great resolution of the public and private map databases that can be accessed on the Internet, especially using the "street-view" version of tools such as Google Earth and Google Maps, software applications that allow to go for a virtual walk in each city, providing highly precise details about the pedestrian actions. The following cities have been chosen: ‐ 11 coastal cities: Alicante, Almería, Bilbao, Cádiz, Castellón de la Plana, Huelva, Las Palmas de Gran Canaria, Palma de Mallorca, Santander, San Sebastián and Tarragona. ‐ 11 interior cities: Albacete, Burgos, Córdoba, León, Lérida, Logroño, Oviedo, Murcia, Pamplona, Valladolid and Vitoria. Finally, as they have the highest percentages regarding the pedestrian areas found in the historic quarters, 3 cities representing a deep pedestrian development have been chosen: Burgos, San Sebastián and Pamplona. Other significant methodological aspects are the many documents found from online sources, especially when preparing the last chapter: “Processes for the pedestrianisation of Spanish cities”, which focuses on the 3 cities chosen. Local newspaper and periodical libraries, official gazettes, the minutes of plenary sessions of councils and the zoning regulation documents (General Zoning Plans, Special Zoning Plans, Municipal Plans and Projects, Mobility Plans, etc.) have been consulted, obtaining accurate information regarding the timeline and the extension of the works carried out. The methodology is supplemented by interviews with key players in the pedestrianisation process, such as the technical staff in councils and the officers drafting and designing the projects, as well as with the direct collaboration by the officers of the councils of the 3 cities chosen, who provided all the information requested. The thesis introduces the pedestrian elements by studying pedestrian areas and existing pedestrianisation processes through the analysis of the literature consulted, providing valuable information for the definition of pedestrian models and showing a brief history and background of the pedestrianisation process. Remarkable papers published by some European experts in the field, such as Carmen Hass-Klau, Rolf Monheim and Collin Buchanan, are covered in the thesis. In Spain, the pedestrianisation actions were late in time in comparison to the rest of Europe, with at least 40 years of delay. The Spanish literature references in this field are limited. The most significant papers are those published by Alfonso Sanz and by Professor Julio Pozueta, with a special mention of one of his last works "La ciudad paseable" (The Walkable City), which describes the pedestrian elements found in the cities and is used a model and an example for this research. Based on the elements above, pedestrian elements are defined as those construction-free public areas that, due to their features and design, ensure the comfortable, convenient use of the spaces, characterised by pedestrian traffic and specifically reserved for this purpose. The thesis detects and specifies the following pedestrian elements: pedestrian streets and squares, pavements (sidewalks) and boulevards with a width exceeding 8 metres, pedestrian promenades, crossing elements (traditional and mechanical), pedestrian spaces linked to new residential areas, colonnades and pedestrian passages or narrow streets with a minimum width of 4 metres, exhibition sites, free spaces and green areas and parks. For each of the 22 cities chosen, a thorough inventory of the elements mentioned above has been made by using worksheets, providing a significant number of individual projects developed in each city. This information is graphically collected and displayed on 2 photomaps, resulting in tools essential for this research. Photomap 1 is made based on the city displayed by Google Earth, making a difference between pedestrian areas and green areas. On Photomap 2, using the tool mentioned above, a difference can be made between persistent and predominant typologies, such as the big pedestrian axes, the local pedestrian networks and singular pedestrian elements. Finally, the 2 photomaps are used in order to establish the so-called "pedestrian footprint", which is highlighted by its attractiveness and appeal, concepts for which the pedestrian field is very useful. With the pedestrian footprint, quick information can be obtained, since the map shows two colours: orange for pedestrian elements (made up of pedestrian areas and green areas) and black for the other non-pedestrian elements. A significant part of these graphic, descriptive documents about each city's pedestrian elements can be found in the thesis appendix. The "Pedestrian Footprint" is regarded in the research as the basic map, obtaining a quick, clear and visual perception of each city's pedestrian space. This footprint provides morphological information regarding the space distribution, the organisation, the balance, the modulation, the cohesion, the pedestrian concentration, etc. in each of the cities chosen. The pedestrian footprint helps solve many of the questions established as the thesis goals, proving that, in general, the pedestrian elements are not organised in the urban plot, as there is no structure modulating the city as a properly linked set of elements. Small networks linked to each other and heterogeneously scattered all over the city are found. It has been verified that the pedestrian elements with the highest concentration and presence in the cities analysed are the pedestrian axes, the historic quarters and the pedestrian elements found in the new residential areas. Besides these 3 elements mentioned, the research analyses in depth other types that are remarkable due to their persistence and repetition in the "Pedestrian Footprint" of each of the 22 cities, such as the pedestrian squares, the colonnades and the singular pedestrian elements: traditional spaces (exhibition sites, passages, historic bridges), new pedestrian elements (pedestrian footbridges, mechanical crossing elements), pedestrian areas around cultural buildings and other cases such as those consisting of recovering and transforming building and urban constructions, intended for a wide range of purposes and of different types, into new pedestrian areas. This work puts light on the definition, location, timeline, proportion, presence and origin, providing results for each of these concepts. At the same time, quantitative information is obtained by measuring the footprint, getting data and percentages on the size of the city, the pedestrian areas and the green areas, the historic quarters and the pedestrian zones, the linear metres of such historic quarters and pedestrian streets. The footprint also provides ratios and rates that are considered as essential in order to better understand the pedestrian elements of each city. A classification of cities is established for each of the areas described, highlighting the cities of Pamplona, San Sebastián and Burgos, as they provide the most positive pedestrian figures and lead most of the classification tables or rankings. According to the criteria considered, the city with the worst values is Alicante, followed by Almería, Palma de Mallorca and Las Palmas de Gran Canaria. In the final chapter in the thesis, the cities of Burgos, San Sebastián and Pamplona are chosen as representative study cases. The aim is to gain more accurate knowledge regarding the pedestrianisation origin, timeline and processes. Each city comprises a chronological sequence made of stages, each of which is thoroughly developed and usually announced as zoning plans or urban development projects, accompanied by the plans of the pedestrian actions. At the end of each stage, data with the figures of the pedestrianisation projects are provided, including a numerical chart with locations, pedestrian area, redeveloped urban areas, pedestrian length and pedestrianisation degree in the historic quarter, as well as the data cumulated throughout each stage. In the crossed conclusions on the three cities, the trend of gradually pedestrianising the historic quarters (even reaching the 100% of the pedestrian streets as in San Sebastián) is verified. However, it is difficult to confirm whether the purpose is to reach an overall pedestrianisation of the historic quarters, since there are many variables that affect and influence each city's pedestrianisation process. In addition, the spread of the pedestrian culture from the internal areas to the external areas, beyond the historic quarters to the nearby expansion areas, is compared. Regarding the origin of pedestrianisations, the thesis comes to the conclusion that there is no initial or theoretical conception from which each city's pedestrian plot is developed. The pedestrian processes implemented are varied and diverse but no fact can be specified as the main cause. Based on the moment and circumstances, the pedestrianisation processes are due to different initiatives promoted by shopkeepers, by the local administrations or by the zoning officers, with their expert contributions to the plans. These three players promote and drive individual pedestrianisation projects that overlap in the course of time. Finally, new lines of research are put forwards, as they can be taken as an additional study and as a supplement to this research, dealing with aspects as significant as the economic, social and mobility factors, closely linked to the pedestrianisation processes, as proven in the literature consulted.

O derramamento do Espírito : fortalecimento dos enfraquecidos

Esta tese se insere dentro da leitura bíblica latino-americana trazendo, nessa ótica, a riqueza e o desafio do que significa ler a Bíblia a partir de um contexto de opressão como é o nosso continente. Nos servimos do roteiro das ciências bíblicas e dos métodos exegéticos modernos. É uma pesquisa bibliográfica onde, eventualmente, consideramos o nível experiêncial, por exemplo quando trabalhamos o tema do derramamento do Espírito nos pentecostais chilenos. Esse capítulo, IV, segue o roteiro da hipótese central de nossa tese na medida que firmamos o princípio de que os pentecostais fazem parte do amplo e diverso grupo dos enfraquecidos como produto de um sistema excludente, baseado na exploração social. Numa situação de crise, o anúncio do derramamento do Espírito é um sinal de salvação, de perspectivas futuras, de conservação e prolongação da vida. Esta tese relê o tema do derramamento do Espírito, a partir de Jl 3,1-5. O tema é analisado num contexto social concreto, onde os enfraquecidos recebem o Espírito do Senhor. O eixo que nos permite fazer esta leitura encontra sua referencial nas pessoas setores sociais - mencionadas como beneficiadas diretas da ação do Espírito. São pessoas que representam setores diferentes; jovens, escravos e escravas, formam o grupo que sustenta e faz funcionar o sistema imperial persa grego - baseado na exploração e mão de obra barata. Eles, juntamente com os idosos que não produzem mais, são a base de sustentação da pirâmide social. Como demostraremos no decorrer da tese, nossa hipótese somente é possível se duas conjunturas se cruzam. Para isso, em num primeiro momento, localizamos e demostramos que o livro de Joel deve ser situado no contexto da literatura apocalíptica (nascimento). Em seguida, demonstramos que o livro de Joel, como produto literário final, deve ser localizado no contexto histórico do pós-exílio, isto é, no tempo dos impérios persa e grego. A confirmação destes dois aspectos desenvolvidos nos capítulos I e II, nos permitem, na análise literária, capítulo III, aprofundar, mediante a análise exegética, e confirmar a nossa hipótese central. Com estes três capítulos demostramos que o derramamento do Espírito do Senhor tem como destinatários preferenciais, senão exclusivos, os setores enfraquecidos pela política social, econômica e religiosa dos impérios persa e grego. Com estes três capítulos desenvolvidos, no quarto capítulo analisamos uma experiência concreta, de um setor majoritariamente pobre que tem se apropriado do texto de Jl 3,1-5, e encontrado nele uma alternativa social, política e religiosa para se manter fiel ao Senhor e por quase um século recria e revive a experiência do derramamento do Espírito. Os velhos, os jovens, os escravos e as escravas, formam o setor dos enfraquecidos. É o setor que nada espera, e que nada terá da parte dos impérios e que, como os pentecostais, pela sua situação de enfraquecimento, acredita que somente uma intervenção externa pode mudar o seu futuro, pode trazer de volta a esperança. Essa intervenção externa começa a ser possível pelo derramamento do Espírito e se concretiza na chegada do dia de Javé, que será grande e terrível, onde o sol e a lua se unem para indicar o monte de Sião e a cidade de Jerusalém como lugar de adoração, refúgio e salvação. Esta perspectiva da ação do Espírito é possível somente no período do pós-exílio. Nesse tempo o Espírito adquire uma dimensão ampla e inclusiva agindo sobre diversos setores sociais, independe de serem ou não judeus.(AU)

Glutamyl-tRNAGln amidotransferase in Deinococcus radiodurans may be confined to asparagine biosynthesis

Asparaginyl-tRNA (Asn-tRNA) and glutaminyl-tRNA (Gln-tRNA) are essential components of protein synthesis. They can be formed by direct acylation by asparaginyl-tRNA synthetase (AsnRS) or glutaminyl-tRNA synthetase (GlnRS). The alternative route involves transamidation of incorrectly charged tRNA. Examination of the preliminary genomic sequence of the radiation-resistant bacterium Deinococcus radiodurans suggests the presence of both direct and indirect routes of Asn-tRNA and Gln-tRNA formation. Biochemical experiments demonstrate the presence of AsnRS and GlnRS, as well as glutamyl-tRNA synthetase (GluRS), a discriminating and a nondiscriminating aspartyl-tRNA synthetase (AspRS). Moreover, both Gln-tRNA and Asn-tRNA transamidation activities are present. Surprisingly, they are catalyzed by a single enzyme encoded by three ORFs orthologous to Bacillus subtilis gatCAB. However, the transamidation route to Gln-tRNA formation is idled by the inability of the discriminating D. radiodurans GluRS to produce the required mischarged Glu-tRNAGln substrate. The presence of apparently redundant complete routes to Asn-tRNA formation, combined with the absence from the D. radiodurans genome of genes encoding tRNA-independent asparagine synthetase and the lack of this enzyme in D. radiodurans extracts, suggests that the gatCAB genes may be responsible for biosynthesis of asparagine in this asparagine prototroph.

Two contact regions between Stat1 and CBP/p300 in interferon γ signaling

Interferon γ (IFN-γ) induces rapid tyrosine phosphorylation of the latent cytoplasmic transcription factor, Stat1, which then forms homodimers, translocates to the nucleus and participates in IFN-γ-induced transcription. However, little is known of the interactions between Stat1 and the general transcription machinery during transcriptional activation. We show here that Stat1 can directly interact with the CREB-binding protein (CBP)/p300 family of transcriptional coactivators. Specifically, two interaction regions were identified: the amino-terminal region of Stat1 interacts with the CREB-binding domain of CBP/p300 and the carboxyl-terminal region of Stat1 interacts with the domain of CBP/p300 that binds adenovirus E1A protein. Transfection experiments suggest a role for these interactions in IFN-γ-induced transcription. Because CBP/p300-binding is required for the adenovirus E1A protein to regulate transcription of many genes during viral replication and cellular transformation, it is possible that the anti-viral effect of IFN-γ is based at least in part on direct competition by nuclear Stat1 with E1A for CBP/p300 binding.

The cellular ecology of progressive neoplastic transformation: A clonal analysis

A comparison was made of the competence for neoplastic transformation in three different sublines of NIH 3T3 cells and multiple clonal derivatives of each. Over 90% of the neoplastic foci produced by an uncloned transformed (t-SA′) subline on a confluent background of nontransformed cells were of the dense, multilayered type, but about half of the t-SA′ clones produced only light foci in assays without background. This asymmetry apparently arose from the failure of the light focus formers to register on a background of nontransformed cells. Comparison was made of the capacity for confluence-mediated transformation between uncloned parental cultures and their clonal derivatives by using two nontransformed sublines, one of which was highly sensitive and the other relatively refractory to confluence-mediated transformation. Transformation was more frequent in the clones than in the uncloned parental cultures for both sublines. This was dramatically so in the refractory subline, where the uncloned culture showed no overt sign of transformation in serially repeated assays but increasing numbers of its clones exhibited progressive transformation. The reason for the greater susceptibility of the pure clones is apparently the suppression of transformation among the diverse membership that makes up the uncloned parental culture. Progressive selection toward increasing degrees of transformation in confluent cultures plays a major role in the development of dense focus formers, but direct induction by the constraint of confluence may contribute by heritably damaging cells. In view of our finding of increased susceptibility to transformation in clonal versus uncloned populations, expansion of some clones at the expense of others during the aging process would contribute to the marked increase of cancer with age.

Phosphatidylinositol 3-kinase signals activation of p70 S6 kinase in situ through site-specific p70 phosphorylation.

The p70 S6 kinase is activated by insulin and mitogens through multisite phosphorylation of the enzyme. One set of activating phosphorylations occurs in a putative autoinhibitory domain in the noncatalytic carboxyl-terminal tail. Deletion of this tail yields a variant (p70 delta CT104) that nevertheless continues to be mitogen regulated. Coexpression with a recombinant constitutively active phosphatidylinositol (PI) 3-kinase (EC 2.7.1.137) gives substantial activation of both full-length p70 and p70 delta CT104 but not Rsk. Activation of p70 delta CT104 by PI 3-kinase and inhibition by wortmannin are each accompanied by parallel and selective changes in the phosphorylation of p70 Thr-252. A Thr or Ser at this site, in subdomain VIII of the catalytic domain just amino-terminal to the APE motif, is necessary for p70 40S kinase activity. The inactive ATP-binding site mutant K123M p70 delta CT104 undergoes phosphorylation of Thr-252 in situ but does not undergo direct phosphorylation by the active PI 3-kinase in vitro. PI 3-kinase provides a signal necessary for the mitogen activation of the p70 S6 kinase, which directs the site-specific phosphorylation of Thr-252 in the p70 catalytic domain, through a distinctive signal transduction pathway.

Estudos sobre a organização genômica, evolução e expressão de microRNAs

Os microRNAs (miRNAs) são pequenos RNAs não codificadores de proteínas presentes na maioria dos eucariotos. Esses RNAs regulam a expressão gênica em nível pós-transcricional através do silenciamento de mRNAs-alvo que possuem sítios complementares às suas sequências, atuando em praticamente todos os processos celulares. Embora a estrutura e função dos miRNAs estejam bem caracterizadas, aspectos relacionados à sua organização genômica, evolução e atuação em doenças são tópicos que apresentam enormes lacunas. Nesta tese, utilizamos abordagens computacionais para investigar estes temas em três trabalhos. No primeiro, processamos e integramos um vasto volume de dados publicamente disponíveis referentes aos miRNAs e genes codificadores de proteínas para cinco espécies de vertebrados. Com isso, construimos uma ferramenta web que permite a fácil inspeção da organização genômica dos miRNAs em regiões inter e intragênicas, o acesso a dados de expressão de miRNAs e de genes codificadores de proteínas (classificados em genes hospedeiros e não hospedeiros de miRNAs), além de outras informações pertinentes. Verificamos que a ferramenta tem sido amplamente utilizada pela comunidade científica e acreditamos que ela possa facilitar a geração de hipóteses associadas à regulação dos miRNAs, principalmente quando estão inseridos em genes hospedeiros. No segundo estudo, buscamos compreender como o contexto genômico e a origem evolutiva dos genes hospedeiros influenciam a expressão e evolução dos miRNAs humanos. Nossos achados mostraram que os miRNAs intragênicos surgem preferencialmente em genes antigos (origem anterior à divergência de vertebrados). Observamos que os miRNAs inseridos em genes antigos têm maior abrangência de expressão do que os inseridos em genes novos. Surpreendentemente, miRNAs jovens localizados em genes antigos são expressos em um maior número de tecidos do que os intergênicos de mesma idade, sugerindo uma vantagem adaptativa inicial que pode estar relacionada com o controle da expressão dos genes hospedeiros, e como consequência, expondo-os a contextos celulares e conjuntos de alvos diversos. Na evolução a longo prazo, vimos que genes antigos conferem maior restrição nos padrões de expressão (menor divergência de expressão) para miRNAs intragênicos, quando comparados aos intergênicos. Também mostramos possíveis associações funcionais relacionadas ao contexto genômico, tais como o enriquecimento da expressão de miRNAs intergênicos em testículo e dos intragênicos em tecidos neurais. Propomos que o contexto genômico e a idade dos genes hospedeiros são fatores-chave para a evolução e expressão dos miRNAs. Por fim, buscamos estabelecer associações entre a expressão diferencial de miRNAs e a quimioresistência em câncer colorretal utilizando linhagens celulares sensíveis e resistentes às drogas 5-Fluoruracil e Oxaliplatina. Dentre os miRNAs identificados, o miR-342 apresentou níveis elevados de expressão nas linhagens sensíveis à Oxaliplatina. Com base na análise dos alvos preditos, detectamos uma significativa associação de miR-342 com a apoptose. A superexpressão de miR-342 na linhagem resistente SW620 evidenciou alterações na expressão de genes da via apoptótica, notavelmente a diminuição da expressão do fator de crescimento PDGFB, um alvo predito possivelmente sujeito à regulação direta pelo miR-342.

The Future Direction of Delaware Law (Including a Brief Exegesis on Fee Shifting Bylaws)

Delaware sets the governance standards for most public companies. The ability to attract corporations could not be explained solely by the existence of a favorable statutory regime. Delaware was not invariably the first or the only state to implement management friendly provisions. Given the interpretive gaps in the statute and the critical importance of the common law in the governance process, courts played an outsized role in setting legal standards. The management friendly nature of the Delaware courts contributed significantly to the state’s attraction to public corporations. A current example of a management friendly trend in the case law had seen the recent decisions setting out the board’s authority to adopt bylaws under Section 109 of the Delaware General Corporation Law (DGCL), particularly those involving the shifting of fees in litigation against the corporation or its directors. The DGCL allows bylaws that address “the business of the corporation, the conduct of its affairs, and its rights or powers or the rights or powers of its stockholders, directors, officers or employees.” The broad parameters are, however, subject to limits. Bylaws cannot be inconsistent with the certificate of incorporation or “the law.” Law includes the common law. The Delaware courts have used the limitations imposed by “the law” to severely restrict the reach of shareholder inspired bylaws. The courts have not used the same principles to impose similar restraints on bylaws adopted by the board of directors. This can be seen with respect to bylaws that restrict or even eliminate the right of shareholders to bring actions against management and the corporation. In ATP Tour, Inc. v. Deutscher Tennis Bund the court approved a fee shifting bylaw that had littl relationship to the internal affairs of the corporation. The decision upheld the bylaw as facially valid.The decision ignored a number of obvious legal infirmities. Among other things, the decision did not adequately address the requirement in Section 109(b) that bylaws be consistent with “the law.” The decision obliquely acknowledged that the provisions would “by their nature, deter litigation” but otherwise made no effort to assess the impact of this deterrence on shareholders causes of action. The provision in fact had the practical effect of restricting, if not eliminating, litigation rights granted by the DGCL and the common law. Perhaps most significantly, however, the bylaws significantly limited common law rights of shareholders to bring actions against the corporation and the board. Given the high dismissal rates for these actions, fee shifting bylaws imposed a meaningful risk of liability on plaintiffs. Moreover, because judgments in derivative suits were paid to the corporation, shareholders serving as plaintiffs confronted the risk of liability without any offsetting direct benefit. By preventing suits in this area, the bylaw effectively insulated the behavior of boards from legal challenge. The ATP decision was poorly reasoned and overstepped acceptable boundaries. The management friendly decision threatened the preeminent role of Delaware in the development of corporate law. The decision raised the specter of federal intervention and the potential for meaningful competition from the states. Because the opinion examined the bylaw in the context of non-stock companies, the reasoning may remain applicable only to those entities and never make the leap to for-profit stock corporations. Nonetheless, the analysis reflects a management friendly approach that does not adequately take into account the impact of the provision on the rights of shareholders.

Ten Years of Judicial Cooperation. Occasional Paper 4/2014

This paper intends to illustrate the respective roles and functions of the Court of Justice of the EU (CJEU) on the one hand, and the Maltese national courts on the other. It will then define the scope and role of the judicial cooperation between the CJEU and the national courts, highlighting the procedure relating to the preliminary rulings. The paper will then briefly describe the cases brought before the CJEU involving Malta, including those concerning requests for preliminary rulings originating from Malta, and the direct actions by the European Commission before the Court of Justice, as well as those before the General Court. After a description of the rationale behind the publication of the book Malta u l-Qorti tal-Ġustizzja tal-Unjoni Ewropea (Malta and the Court of Justice of the European Union), and following the conference in which it was presented, the main points that emerged from the conference will serve as a backdrop to some statistical analysis pertaining to the Maltese cases, as well as some reflections on the current situation of the judicial cooperation obtained after ten years. It will propose that, besides a mere statistical analysis of the raw figures that emerge, one must rather address his attention to the spirit of EU membership, and reflect on whether Malta’s legal system has actually absorbed and understood the full meaning of the EU membership, ten years after it took place.

A Spatial Analysis of Agricultural Land Prices in Bavaria. Factor Markets Working Document No. 50, June 2013

This paper empirically analyses a dataset of more than 7,300 agricultural land sales transactions from 2001 and 2007 to identify the factors influencing agricultural land prices in Bavaria. We use a general spatial model, which combines a spatial lag and a spatial error model, and in addition account for endogeneity introduced by the spatially lagged dependent variable as well as other explanatory variables. Our findings confirm the strong influence of agricultural factors such as land productivity, of variables describing the regional land market structure, and of non-agricultural factors such as urban pressure on agricultural land prices. Moreover, the involvement of public authorities as a seller or buyer increases sales prices in Bavaria. We find a significant capitalisation of government support payments into agricultural land, where a decrease of direct payments by 1% would decrease land prices in 2007 and 2001 by 0.27% and 0.06%, respectively. In addition, we confirm strong spatial relationships in our dataset. Neglecting this leads to biased estimates, especially if aggregated data is used. We find that the price of a specific plot increases by 0.24% when sales prices in surrounding areas increase by 1%.

Organic geochemistry of sediments at DSDP Legs 56-57 Holes

The quantity, type, and maturity of the organic matter of Quaternary and Tertiary sediments from the Japan Trench (DSDP Leg 56, Sites 434 and 436; and Leg 57, Sites 438, 439 and 440) were determined. The hydrocarbons in lipid extracts were analyzed by capillary- column gas chromatography and combined gas chromatography/ mass spectrometry. Kerogen concentrates were investigated by microscopy, and vitrinite-reflectance values were determined. Measured organic-carbon values were in the range of 0.13 to 1.00 per cent. Extract yields, however, were extremely low. Normalized to organic carbon, total extracts ranged from 4.1 to 15.7 mg/g Corg. Gas chromatography of non-aromatic hydrocarbons showed that all sediments, except one Oligocene sample, contained very immature, mainly terrigenous organic material. This was indicated by n-alkane maxima at C29 and C31 and high odd-carbon-number predominances. Unsaturated steroid hydrocarbons were found to be major cyclic compounds in lower- and middle-Miocene samples from the upper inner trench slope (Sites 438 and 439). Perylene was the dominating aromatic hydrocarbon in all but the Oligocene sample. Microscopy showed kerogens rich in terrigenous organic particles, with a major portion of recycled vitrinite. Nevertheless, almost all the liptinite particles appeared to be primary. This is a paradox, as the bulk of the samples were composed of hemipelagic mineral matter with a major siliceous biogenic (planktonic) component. A trend of reduced size and increased roundness can be seen for the vitrinite/ inertinite particles from west to east (from upper inner trench slope to outer trench slope). All sediments but one are relatively immature, with mean huminite-reflectance values (Ro)in the range of 0.30 to 0.45 per cent. The oldest and deepest sediment investigated, an Oligocene sandstone from Site 439, yielded a mean vitrinitereflectance value of 0.74 per cent and a mature n-alkane distribution. This sample may indicate a geothermal event in late Oligocene time. It failed to affect the overlying lower Miocene and may have been caused by an intrusion. Boulders of acidic igneous rocks in the Oligocene can be interpreted as witnesses of nearby volcanic activity accompanied by intrusions.