(Table 1) Concentrations of neutral and phenolic organohalogen compounds in adipose tissue, blood, brain and liver of polar bears (Ursus maritimus) from East Greenland

The tissue-specific composition of sum classes of brominated and chlorinated contaminants and metabolic/degradation byproducts was determined in adult male and female polar bears from East Greenland. Significantly (p < 0.05) higher concentrations of SUM-PCBs, various other organochlorines such as SUM-CHL, p,p'-DDE, SUM-CBz, SUM-HCHs, octachlorostyrene (OCS),SUM-mirex, dieldrin, the flame retardants SUM-PBDEs, and total-(R)-hexabromocyclododecane (HBCD), SUM-methylsulfonyl (MeSO2)-PCBs and 3-MeSO2-p,p'-DDE, were found in the adipose and liver tissues relative to whole blood and brain. In contrast, SUM-hydroxyl (OH)-PCB, 4-OH-heptachlorostyrene and SUM-OH-PBDE concentrations were significantly highest (p < 0.05) in whole blood, whereas the highest concentrations of SUM-OH-PBBs were found in the adipose tissue. Based on the total concentrations of all organohalogens in all three tissues and blood, the combined body burden was estimated to be 1.34 ± 0.12 g, where >91% of this amount was accounted for by the adipose tissue alone, followed by the liver, whole blood, and brain. These results show that factors such as protein association and lipid solubility appear to be differentially influencing the toxicokinetics, in terms of tissue composition/localization and burden, of organohalogen classes with respect to chemical structure and properties such as the type of halogenation (e.g., chlorination or bromination), and the presence or absence of additional phenyl group substituents (e.g., MeO and OH groups). The tissue- and blood-specific accumulation (or retention) among organohalogen classes indicates that exposure and any potential contaminant-mediated effects in these polar bears are likely tissue or blood specific.

Ecological assessment data of the marine flora and fauna of Point Lookout in 2014 (PLEA)

In 2014, UniDive (The University of Queensland Underwater Club) conducted an ecological assessment of the Point Lookout Dive sites for comparison with similar surveys conducted in 2001. Involvement in the project was voluntary. Members of UniDive who were marine experts conducted training for other club members who had no, or limited, experience in identifying marine organisms and mapping habitats. Since the 2001 detailed baseline study, no similar seasonal survey has been conducted. The 2014 data is particularly important given that numerous changes have taken place in relation to the management of, and potential impacts on, these reef sites. In 2009, Moreton Bay Marine Park was re-zoned, and Flat Rock was converted to a marine national park zone (Green zone) with no fishing or anchoring. In 2012, four permanent moorings were installed at Flat Rock. Additionally, the entire area was exposed to the potential effects of the 2011 and 2013 Queensland floods, including flood plumes which carried large quantities of sediment into Moreton Bay and surrounding waters. The population of South East Queensland has increased from 2.49 million in 2001 to 3.18 million in 2011 (BITRE, 2013). This rapidly expanding coastal population has increased the frequency and intensity of both commercial and recreational activities around Point Lookout dive sites (EPA 2008). Methodology used for the PLEA project was based on the 2001 survey protocols, Reef Check Australia protocols and Coral Watch methods. This hybrid methodology was used to monitor substrate and benthos, invertebrates, fish, and reef health impacts. Additional analyses were conducted with georeferenced photo transects. The PLEA marine surveys were conducted over six weekends in 2014 totaling 535 dives and 376 hours underwater. Two training weekends (February and March) were attended by 44 divers, whilst biological surveys were conducted on seasonal weekends (February, May, July and October). Three reefs were surveyed, with two semi-permanent transects at Flat Rock, two at Shag Rock, and one at Manta Ray Bommie. Each transect was sampled once every survey weekend, with the transect tapes deployed at a depth of 10 m below chart datum. Fish populations were assessed using a visual census along 3 x 20 m transects. Each transect was 5 m wide (2.5 m either side of the transect tape), 5 m high and 20 m in length. Fish families and species were chosen that are commonly targeted by recreational or commercial fishers, or targeted by aquarium collectors, and that were easily identified by their body shape. Rare or otherwise unusual species were also recorded. Target invertebrate populations were assessed using visual census along 3 x 20 m transects. Each transect was 5 m wide (2.5 m either side of the transect tape) and 20 m in length. The diver surveying invertebrates conducted a 'U-shaped' search pattern, covering 2.5 m on either side of the transect tape. Target impacts were assessed using a visual census along the 3 x 20 m transects. Each transect was 5 m wide (2.5 m either side of the transect tape) and 20 m in length. The transect was surveyed via a 'U-shaped' search pattern, covering 2.5 m on either side of the transect tape. Substrate surveys were conducted using the point sampling method, enabling percentage cover of substrate types and benthic organisms to be calculated. The substrate or benthos under the transect line was identified at 0.5m intervals, with a 5m gap between each of the three 20m segments. Categories recorded included various growth forms of hard and soft coral, key species/growth forms of algae, other living organisms (i.e. sponges), recently killed coral, and, non-living substrate types (i.e. bare rock, sand, rubble, silt/clay).

Seawater carbonate chemistry and processes during experiments with crabs Chionoecetes tanneri and Cancer magister, 2007

Rising levels of atmospheric carbon dioxide could be curbed by large-scale sequestration of CO2 in the deep sea. Such a solution requires prior assessment of the impact of hypercapnic, acidic seawater on deep-sea fauna. Laboratory studies were conducted to assess the short-term hypercapnic tolerance of the deep-sea Tanner crab Chionoecetes tanneri, collected from 1000 m depth in Monterey Canyon off the coast of central California, USA. Hemolymph acid- base parameters were monitored over 24 h of exposure to seawater equilibrated with ~1% CO2 (seawater PCO2 ~6 torr or 0.8 kPa, pH 7.1), and compared with those of the shallow-living Dungeness crab Cancer magister. Short-term hypercapnia-induced acidosis in the hemolymph of Chionoecetes tanneri was almost uncompensated, with a net 24 h pH reduction of 0.32 units and a net bicarbonate accumulation of only 3 mM. Under simultaneous hypercapnia and hypoxia, short-term extracellular acidosis in Chionoecetes tanneri was completely uncompensated. In contrast, Cancer magister fully recovered its hemolymph pH over 24 h of hypercapnic exposure by net accumulation of 12 mM bicarbonate from the surrounding medium. The data support the hypothesis that deep-sea animals, which are adapted to a stable environment and exhibit reduced metabolic rates, lack the short-term acid-base regulatory capacity to cope with the acute hypercapnic stress that would accompany large-scale CO2 sequestration. Additionally, the data indicate that sequestration in oxygen-poor areas of the ocean would be even more detrimental to deep-sea fauna.

Safety, feasibility, and long-term results of percutaneous closure of atrial septal defects using the Amplatzer septal occluder without periprocedural echocardiography.

OBJECTIVES We sought to assess the safety and efficacy of percutaneous closure of atrial septal defects (ASDs) under fluoroscopic guidance only, without periprocedural echocardiographic guidance. BACKGROUND Percutaneous closure of ASDs is usually performed using simultaneous fluoroscopic and transthoracic, transesophageal (TEE), or intracardiac echocardiographic (ICE) guidance. However, TEE requires deep sedation or general anesthesia, which considerably lengthens the procedure. TEE and ICE increase costs. METHODS Between 1997 and 2008, a total of 217 consecutive patients (age, 38 ± 22 years; 155 females and 62 males), of whom 44 were children ≤16 years, underwent percutaneous ASD closure with an Amplatzer ASD occluder (AASDO). TEE guidance and general anesthesia were restricted to the children, while devices were implanted under fluoroscopic guidance only in the adults. For comparison of technical safety and feasibility of the procedure without echocardiographic guidance, the children served as a control group. RESULTS The implantation procedure was successful in all but 3 patients (1 child and 2 adults; 1.4%). Mean device size was 23 ± 8 mm (range, 4-40 mm). There was 1 postprocedural complication (0.5%; transient perimyocarditis in an adult patient). At last echocardiographic follow-up, 13 ± 23 months after the procedure, 90% of patients had no residual shunt, whereas a minimal, moderate, or large shunt persisted in 7%, 1%, and 2%, respectively. Four adult patients (2%) underwent implantation of a second device for a residual shunt. During a mean follow-up period of 3 ± 2 years, 2 deaths and 1 ischemic stroke occurred. CONCLUSION According to these results, percutaneous ASD closure using the AASDO without periprocedural echocardiographic guidance seems safe and feasible.

The influence of ocean acidification on nitrogen regeneration and nitrous oxide production in the northwest European shelf sea

The assimilation and regeneration of dissolved inorganic nitrogen, and the concentration of N2O, was investigated at stations located in the NW European shelf sea during June/July 2011. These observational measurements within the photic zone demonstrated the simultaneous regeneration and assimilation of NH4+, NO2- and NO3-. NH4+ was assimilated at 1.82-49.12 nmol N/L/h and regenerated at 3.46-14.60 nmol N/L/h; NO2- was assimilated at 0-2.08 nmol N/L/h and regenerated at 0.01-1.85 nmol N/L/h; NO3-was assimilated at 0.67-18.75 nmol N/L/h and regenerated at 0.05-28.97 nmol N/L/h. Observations implied that these processes were closely coupled at the regional scale and that nitrogen recycling played an important role in sustaining phytoplankton growth during the summer. The [N2O], measured in water column profiles, was 10.13 ± 1.11 nmol/L and did not strongly diverge from atmospheric equilibrium indicating that sampled marine regions were neither a strong source nor sink of N2O to the atmosphere. Multivariate analysis of data describing water column biogeochemistry and its links to N-cycling activity failed to explain the observed variance in rates of N-regeneration and N-assimilation, possibly due to the limited number of process rate observations. In the surface waters of five further stations, ocean acidification (OA) bioassay experiments were conducted to investigate the response of NH4+ oxidising and regenerating organisms to simulated OA conditions, including the implications for [N2O]. Multivariate analysis was undertaken which considered the complete bioassay data set of measured variables describing changes in N-regeneration rate, [N2O] and the biogeochemical composition of seawater. While anticipating biogeochemical differences between locations, we aimed to test the hypothesis that the underlying mechanism through which pelagic N-regeneration responded to simulated OA conditions was independent of location. Our objective was to develop a mechanistic understanding of how NH4+ regeneration, NH4+ oxidation and N2O production responded to OA. Results indicated that N-regeneration process responses to OA treatments were location specific; no mechanistic understanding of how N-regeneration processes respond to OA in the surface ocean of the NW European shelf sea could be developed.

Challenges for the Multilingual Web of Data

The Web has witnessed an enormous growth in the amount of semantic information published in recent years. This growth has been stimulated to a large extent by the emergence of Linked Data. Although this brings us a big step closer to the vision of a Semantic Web, it also raises new issues such as the need for dealing with information expressed in different natural languages. Indeed, although the Web of Data can contain any kind of information in any language, it still lacks explicit mechanisms to automatically reconcile such information when it is expressed in different languages. This leads to situations in which data expressed in a certain language is not easily accessible to speakers of other languages. The Web of Data shows the potential for being extended to a truly multilingual web as vocabularies and data can be published in a language-independent fashion, while associated language-dependent (linguistic) information supporting the access across languages can be stored separately. In this sense, the multilingual Web of Data can be realized in our view as a layer of services and resources on top of the existing Linked Data infrastructure adding i) linguistic information for data and vocabularies in different languages, ii) mappings between data with labels in different languages, and iii) services to dynamically access and traverse Linked Data across different languages. In this article we present this vision of a multilingual Web of Data. We discuss challenges that need to be addressed to make this vision come true and discuss the role that techniques such as ontology localization, ontology mapping, and cross-lingual ontology-based information access and presentation will play in achieving this. Further, we propose an initial architecture and describe a roadmap that can provide a basis for the implementation of this vision.

Environmental variables modeling based on remote sensing data using time series analysis: forestry and agricultural applications

En la actualidad, el seguimiento de la dinámica de los procesos medio ambientales está considerado como un punto de gran interés en el campo medioambiental. La cobertura espacio temporal de los datos de teledetección proporciona información continua con una alta frecuencia temporal, permitiendo el análisis de la evolución de los ecosistemas desde diferentes escalas espacio-temporales. Aunque el valor de la teledetección ha sido ampliamente probado, en la actualidad solo existe un número reducido de metodologías que permiten su análisis de una forma cuantitativa. En la presente tesis se propone un esquema de trabajo para explotar las series temporales de datos de teledetección, basado en la combinación del análisis estadístico de series de tiempo y la fenometría. El objetivo principal es demostrar el uso de las series temporales de datos de teledetección para analizar la dinámica de variables medio ambientales de una forma cuantitativa. Los objetivos específicos son: (1) evaluar dichas variables medio ambientales y (2) desarrollar modelos empíricos para predecir su comportamiento futuro. Estos objetivos se materializan en cuatro aplicaciones cuyos objetivos específicos son: (1) evaluar y cartografiar estados fenológicos del cultivo del algodón mediante análisis espectral y fenometría, (2) evaluar y modelizar la estacionalidad de incendios forestales en dos regiones bioclimáticas mediante modelos dinámicos, (3) predecir el riesgo de incendios forestales a nivel pixel utilizando modelos dinámicos y (4) evaluar el funcionamiento de la vegetación en base a la autocorrelación temporal y la fenometría. Los resultados de esta tesis muestran la utilidad del ajuste de funciones para modelizar los índices espectrales AS1 y AS2. Los parámetros fenológicos derivados del ajuste de funciones permiten la identificación de distintos estados fenológicos del cultivo del algodón. El análisis espectral ha demostrado, de una forma cuantitativa, la presencia de un ciclo en el índice AS2 y de dos ciclos en el AS1 así como el comportamiento unimodal y bimodal de la estacionalidad de incendios en las regiones mediterránea y templada respectivamente. Modelos autorregresivos han sido utilizados para caracterizar la dinámica de la estacionalidad de incendios y para predecir de una forma muy precisa el riesgo de incendios forestales a nivel pixel. Ha sido demostrada la utilidad de la autocorrelación temporal para definir y caracterizar el funcionamiento de la vegetación a nivel pixel. Finalmente el concepto “Optical Functional Type” ha sido definido, donde se propone que los pixeles deberían ser considerados como unidades temporales y analizados en función de su dinámica temporal. ix SUMMARY A good understanding of land surface processes is considered as a key subject in environmental sciences. The spatial-temporal coverage of remote sensing data provides continuous observations with a high temporal frequency allowing the assessment of ecosystem evolution at different temporal and spatial scales. Although the value of remote sensing time series has been firmly proved, only few time series methods have been developed for analyzing this data in a quantitative and continuous manner. In the present dissertation a working framework to exploit Remote Sensing time series is proposed based on the combination of Time Series Analysis and phenometric approach. The main goal is to demonstrate the use of remote sensing time series to analyze quantitatively environmental variable dynamics. The specific objectives are (1) to assess environmental variables based on remote sensing time series and (2) to develop empirical models to forecast environmental variables. These objectives have been achieved in four applications which specific objectives are (1) assessing and mapping cotton crop phenological stages using spectral and phenometric analyses, (2) assessing and modeling fire seasonality in two different ecoregions by dynamic models, (3) forecasting forest fire risk on a pixel basis by dynamic models, and (4) assessing vegetation functioning based on temporal autocorrelation and phenometric analysis. The results of this dissertation show the usefulness of function fitting procedures to model AS1 and AS2. Phenometrics derived from function fitting procedure makes it possible to identify cotton crop phenological stages. Spectral analysis has demonstrated quantitatively the presence of one cycle in AS2 and two in AS1 and the unimodal and bimodal behaviour of fire seasonality in the Mediterranean and temperate ecoregions respectively. Autoregressive models has been used to characterize the dynamics of fire seasonality in two ecoregions and to forecasts accurately fire risk on a pixel basis. The usefulness of temporal autocorrelation to define and characterized land surface functioning has been demonstrated. And finally the “Optical Functional Types” concept has been proposed, in this approach pixels could be as temporal unities based on its temporal dynamics or functioning.

Overall and local mechanical behaviour of dissimilar laser welded joints

The present investigation addresses the overall and local mechanical performance of dissimilar joints of low carbon steel (CS) and stainless steel (SS) thin sheets achieved by laser welding in case of heat source displacement from the weld gap centreline towards CS. Microstructure characterization and residua! strain scanning, carried out by neutron diffraction, were used to assess the joints features. It was found that the heat source position influences the base metals dilution and the residua! stress field associated to the welding process; the transverse residual stress is smaller than for the longitudinal component, of magnitudes close to the parent CS yield strength. Furthermore, compressive transverse residual stresses were encountered at the SS-weld interface. The tensile behavior of the joint different zones assessed by using a video-image based system (VIC-2D) reveals that the residual stress field, together with the positive difference in yield between the weld metal and the base materials protects the joint from being piastically deformed. The tensile loadings of flat transverse specimens generate the strain localization and failure in CS, far away from the weld.En este trabajo se exponen los resultados de una investigacion sobre el comportamiento mecanico de soldaduras disimiles acero inoxidable-acero al carbono, realizadas para unir chapas delgadas, desplazando la fuente de calor del eje longitudinal de la union soldada por laser sobre el acero al carbono. Se han determinado las caracteristicas microestructurales de la union soldada, las tensiones residuales generadas (mediante difraccion de neutrones) y las curvas tension-deformacion locales y globales, mediante medidas locales de deformacion empleando el sistema VIC-2D "video image correlation". El desplazamiento de la fuente de calor infiuye en la dilution de los metales base y el campo de tensiones residuales asociado al proceso de soldeo; las tensiones residuales medidas en direction longitudinal se aproximan al limite elastico del acero al carbono, mientras que las tensiones residuales transversales son menores, e incluso de compresion. El ensayo a traccion de la union soldada revela que las tensiones residuales y la diferencia de limite elastico entre los metales base y la soldadura propician que la rotura se produzca por inestabilidad plastica del acero al carbono, lejos de la soldadura, sin que la union plastifique.

The Spreading of Christianity and the introduction of Modern Architecture in Shannxi, China (1840-1949): Christian churches and traditional Chinese architecture

La arquitectura china ha experimentado grandes cambios a lo largo de un extenso proceso histórico. El hito de mayor importancia es el que da paso al denominado Tiempo Moderno, periodo en el cual irrumpe por vez primera en China la arquitectura occidental, que comienza a tener una influencia muy activa y significativa sobre los rasgos y la identidad de la arquitectura tradicional china, hasta ese momento el único estilo o forma de hacer –muy diferente, en cuanto a su concepción y fisonomía, de los planteamientos occidentales- que había sobrevivido sin desvíos significativos, configurando un panorama milenario bastante homogéneo en los aspectos técnicos y artísticos en el desarrollo de esa arquitectura. Por un cúmulo de complejas razones, la mayor parte de la arquitectura china del periodo feudal -es decir el que forman todos los años anteriores a 1849- ha desaparecido. Sin embargo, desde la fecha indicada hasta la Revolución de 1949 (el denominado periodo semicolonial o semifeudal), sí se conservan muchas edificaciones, que fueron mejor construidas y mantenidas luego, destacando por su importancia en ese sentido las iglesias cristianas. Dichos templos representan cronológicamente, no sólo la primera irrupción de la arquitectura clásica occidental en China, sino el inicio de un proceso de modernización de la profundamente enraizada y, en buena medida, estancada arquitectura vernácula, combinando técnicas y estilos de ambos planteamientos, para dar como resultado originales edificaciones de un singular eclecticismo que caracterizarían buena parte de la arquitectura de dicha etapa semicolonial. En términos generales, últimamente se ha ido prestando cada vez más atención a esta arquitectura de los tiempos modernos, aunque las iglesias cristianas de la provincia de Shaanxi no han sido objeto de estudio específico, a pesar de que su tipología es muy representativa de las construcciones de esta clase en otras regiones del interior de China. La investigación que desarrolla la presente tesis doctoral sale al paso de esa deficiencia, abriendo puertas a la continuación del trabajo referido, extendido a otras zonas o arquitecturas, y, por extensión, a la profundización analítica de la hibridación arquitectónica y cultural entre China y Occidente. Sobre las bases de investigación documental, estudios de campo y dibujo, la tesis plantea un estudio aclaratorio de los rasgos y raíces de la arquitectura tradicional china, al que sigue otro histórico y tipológico de los templos cristianos en la provincia de Shaanxi, deteniéndose en sus características fundamentales, situación (uso) actual y estado de conservación. Se ha considerado imprescindible añadir al trabajo, como apéndice, un elaborado glosario conceptual ilustrado de términos básicos arquitectónicos y constructivos, en chino, inglés y español. ABSTRACT The Chinese architecture has gone through great changes during the long process of history. The tremendous changing period was the named Modern Times of China when, for the very first time, the western architecture was introduced into China and became to influence majorly on the traditional Chinese architecture. Before that, the traditional Chinese architecture which has its own, yet totally different system from the occidental architecture system was the only architectural style could be found in China. Although, due to many historical, conceptual and architectural characteristic reasons, large amount of the ancient Chinese architecture built in the feudal China was not preserved, there are a lot of buildings of semi-feudal China that was well constructed and conserved. The most important architectural type of the semi-feudal China is the Christian Churches. It was not only the first western architectural form that was brought into and well developed in China, but also was the beginner of the modernization process of Chinese architecture. Because of the deep root of the 2000-year traditional Chinese architecture, all the Christian Churches built in China during the semi-colonial society has a combined style of both the traditional Chinese architecture and the classic western churches. They are a priceless asset of the Chinese architectural history. Recently, more and more attention had been paid on the Chinese Modern Times architecture, however, the Christian Churches in Shaanxi Province, the province which has a unique history with the Christian, but less economically developed have never been researched yet. The Christian Churches of Shaanxi Province reflect the general feature of developing history of the Christian Churches of common inner-land regions in China. The research opens the door to further study on other Christian Churches and related buildings, and also for the further study on the Chinese-western architectural and culture communication. On the base of document research, field survey and mapping, in this thesis, an in-depth study had been done on the general history of the features and roots of the traditional Chinese architecture, the developing history of the Christian Churches of Shaanxi Province and the architectural types, examples, characteristics, present situation and conservation status. By comparing the Christian Churches of the cities in Shaanxi province to the Christian Churches in other more developed cities, and by comparing the Christian Churches in China to the classic western churches, the architectural combination feature of the Christian Churches in China are highlighted. The thesis is a fundamental research on which many further studies about the architectural developing history, characteristics and conservation of the Christian Churches in China could be done. It is considered essential to add to the work, as an appendix, an elaborate conceptual illustrated glossary of architectural and construction terms in Chinese, English and Spanish.

Some advances in extensive bridge monitoring using low cost dynamic characterization

Dynamic measurements will become a standard for bridge monitoring in the near future. This fact will produce an important cost reduction for maintenance. US Administration has a long term intensive research program in order to diminish the estimated current maintenance cost of US$7 billion per year over 20 years. An optimal intervention maintenance program demands a historical dynamical record, as well as an updated mathematical model of the structure to be monitored. In case that a model of the structure is not actually available it is possible to produce it, however this possibility does not exist for missing measurement records from the past. Current acquisition systems to monitor structures can be made more efficient by introducing the following improvements, under development in the Spanish research Project “Low cost bridge health monitoring by ambient vibration tests using wireless sensors”: (a) a complete wireless system to acquire sensor data, (b) a wireless system that permits the localization and the hardware identification of the whole sensor system. The applied localization system has been object of a recent patent, and (c) automatization of the modal identification process, aimed to diminish human intervention. This system is assembled with cheap components and allows the simultaneous use of a large number of sensors at a low placement cost. The engineer’s intervention is limited to the selection of sensor positions, probably based on a preliminary FE analysis. In case of multiple setups, also the position of a number of fixed reference sensors has to be decided. The wireless localization system will obtain the exact coordinates of all these sensors positions. When the selection of optimal positions is difficult, for example because of the lack of a proper FE model, this can be compensated by using a higher number of measuring (also reference) points. The described low cost acquisition system allows the responsible bridge administration to obtain historical dynamic identification records at reasonable costs that will be used in future maintenance programs. Therefore, due to the importance of the baseline monitoring record of a new bridge, a monitoring test just after its construction should be highly recommended, if not compulsory.

Cycling Dof Factors: Molecular and functional characterization of Arabidopsis thaliana AtCDF3 and tomato (Solanum lycopersicum L.) SlCDF3 in response to abiotic stress

El tomate (Solanum lycopersicum L.) es considerado uno de los cultivos hortícolas de mayor importancia económica en el territorio Español. Sin embargo, su producción está seriamente afectada por condiciones ambientales adversas como, salinidad, sequía y temperaturas extremas. Para resolver los problemas que se presentan en condiciones de estrés, se han empleado una serie de técnicas culturales que disminuyen sus efectos negativos, siendo de gran interés el desarrollo de variedades tolerantes. En este sentido la obtención y análisis de plantas transgénicas, ha supuesto un avance tecnológico, que ha facilitado el estudio y la evaluación de genes seleccionados en relación con la tolerancia al estrés. Estudios recientes han mostrado que el uso de genes reguladores como factores de transcripción (FTs) es una gran herramienta para obtener nuevas variedades de tomate con mayor tolerancia a estreses abióticos. Las proteínas DOF (DNA binding with One Finger) son una familia de FTs específica de plantas (Yangisawa, 2002), que están involucrados en procesos fisiológicos exclusivos de plantas como: asimilación del nitrógeno y fijación del carbono fotosintético, germinación de semilla, metabolismo secundario y respuesta al fotoperiodo pero su preciso rol en la tolerancia a estrés abiótico se desconoce en gran parte. El trabajo descrito en esta tesis tiene como objetivo estudiar genes reguladores tipo DOF para incrementar la tolerancia a estrés abiotico tanto en especies modelo como en tomate. En el primer capítulo de esta tesis se muestra la caracterización funcional del gen CDF3 de Arabidopsis, así como su papel en la respuesta a estrés abiótico y otros procesos del desarrollo. La expresión del gen AtCDF3 es altamente inducido por sequía, temperaturas extremas, salinidad y tratamientos con ácido abscísico (ABA). La línea de inserción T-DNA cdf3-1 es más sensible al estrés por sequía y bajas temperaturas, mientras que líneas transgénicas de Arabidopsis 35S::AtCDF3 aumentan la tolerancia al estrés por sequía, osmótico y bajas temperaturas en comparación con plantas wild-type (WT). Además, estas plantas presentan un incremento en la tasa fotosintética y apertura estomática. El gen AtCDF3 se localiza en el núcleo y que muestran una unión específica al ADN con diferente afinidad a secuencias diana y presentan diversas capacidades de activación transcripcional en ensayos de protoplastos de Arabidopsis. El dominio C-terminal de AtCDF3 es esencial para esta localización y su capacidad activación, la delección de este dominio reduce la tolerancia a sequía en plantas transgénicas 35S::AtCDF3. Análisis por microarray revelan que el AtCDF3 regula un set de genes involucrados en el metabolismo del carbono y nitrógeno. Nuestros resultados demuestran que el gen AtCDF3 juega un doble papel en la regulación de la respuesta a estrés por sequía y bajas temperaturas y en el control del tiempo de floración. En el segundo capítulo de este trabajo se lleva a cabo la identificación de 34 genes Dof en tomate que se pueden clasificar en base a homología de secuencia en cuatro grupos A-D, similares a los descritos en Arabidopsis. Dentro del grupo D se han identificado cinco genes DOF que presentan características similares a los Cycling Dof Factors (CDFs) de Arabidopsis. Estos genes son considerados ortólogos de Arabidopsis CDF1-5, y han sido nombrados como Solanum lycopersicum CDFs o SlCDFs. Los SlCDF1-5 son proteínas nucleares que muestran una unión específica al ADN con diferente afinidad a secuencias diana y presentan diversas capacidades de activación transcripcional in vivo. Análisis de expresión de los genes SlCDF1-5 muestran diferentes patrones de expresión durante el día y son inducidos de forma diferente en respuesta a estrés osmótico, salino, y de altas y bajas temperaturas. Plantas de Arabidopsis que sobre-expresan SlCDF1 y SlCDF3 muestran un incremento de la tolerancia a la sequía y salinidad. Además, de la expresión de varios genes de respuesta estrés como AtCOR15, AtRD29A y AtERD10, son expresados de forma diferente en estas líneas. La sobre-expresión de SlCDF3 en Arabidopsis promueve un retardo en el tiempo de floración a través de la modulación de la expresión de genes que controlan la floración como CONSTANS (CO) y FLOWERING LOCUS T (FT). En general, nuestros datos demuestran que los SlCDFs están asociados a funciones aun no descritas, relacionadas con la tolerancia a estrés abiótico y el control del tiempo de floración a través de la regulación de genes específicos y a un aumento de metabolitos particulares. ABSTRACT Tomato (Solanum lycopersicum L.) is one of the horticultural crops of major economic importance in the Spanish territory. However, its production is being affected by adverse environmental conditions such as salinity, drought and extreme temperatures. To resolve the problems triggered by stress conditions, a number of agricultural techniques that reduce the negative effects of stress are being frequently applied. However, the development of stress tolerant varieties is of a great interest. In this direction, the technological progress in obtaining and analysis of transgenic plants facilitated the study and evaluation of selected genes in relation to stress tolerance. Recent studies have shown that a use of regulatory genes such as transcription factors (TFs) is a great tool to obtain new tomato varieties with greater tolerance to abiotic stresses. The DOF (DNA binding with One Finger) proteins form a family of plant-specific TFs (Yangisawa, 2002) that are involved in the regulation of particular plant processes such as nitrogen assimilation, photosynthetic carbon fixation, seed germination, secondary metabolism and flowering time bur their precise roles in abiotic stress tolerance are largely unknown. The work described in this thesis aims at the study of the DOF type regulatory genes to increase tolerance to abiotic stress in both model species and the tomato. In the first chapter of this thesis, we present molecular characterization of the Arabidopsis CDF3 gene as well as its role in the response to abiotic stress and in other developmental processes. AtCDF3 is highly induced by drought, extreme temperatures, salt and abscisic acid (ABA) treatments. The cdf3-1 T-DNA insertion mutant was more sensitive to drought and low temperature stresses, whereas the AtCDF3 overexpression enhanced the tolerance of transgenic plants to drought, cold and osmotic stress comparing to the wild-type (WT) plants. In addition, these plants exhibit increased photosynthesis rates and stomatal aperture. AtCDF3 is localized in the nuclear region, displays specific binding to the canonical DNA target sequences and has a transcriptional activation activity in Arabidopsis protoplast assays. In addition, the C-terminal domain of AtCDF3 is essential for its localization and activation capabilities and the deletion of this domain significantly reduces the tolerance to drought in transgenic 35S::AtCDF3 overexpressing plants. Microarray analysis revealed that AtCDF3 regulated a set of genes involved in nitrogen and carbon metabolism. Our results demonstrate that AtCDF3 plays dual roles in regulating plant responses to drought and low temperature stress and in control of flowering time in vegetative tissues. In the second chapter this work, we carried out to identification of 34 tomato DOF genes that were classified by sequence similarity into four groups A-D, similar to the situation in Arabidopsis. In the D group we have identified five DOF genes that show similar characteristics to the Cycling Dof Factors (CDFs) of Arabidopsis. These genes were considered orthologous to the Arabidopsis CDF1 - 5 and were named Solanum lycopersicum CDFs or SlCDFs. SlCDF1-5 are nuclear proteins that display specific binding to canonical DNA target sequences and have transcriptional activation capacities in vivo. Expression analysis of SlCDF1-5 genes showed distinct diurnal expression patterns and were differentially induced in response to osmotic, salt and low and high temperature stresses. Arabidopsis plants overexpressing SlCDF1 and SlCDF3 showed increased drought and salt tolerance. In addition, various stress-responsive genes, such as AtCOR15, AtRD29A and AtERD10, were expressed differently in these lines. The overexpression of SlCDF3 in Arabidopsis also results in the late flowering phenotype through the modulation of the expression of flowering control genes such CONSTANS (CO) and FLOWERING LOCUS T (FT). Overall, our data connet SlCDFs to undescribed functions related to abiotic stress tolerance and flowering time through the regulation of specific target genes and an increase in particular metabolites.

Detecting, Localizing and Following Dynamic Objects with a Mini-UAV

This paper presents an approach for the detection, localization and following of dynamic terrestrial objects using a mini-UAV. The development is intended to be used for surveillance of large infrastructures. The detection algorithm is based on finding several pre-defined characteristics of the target, such as color, shape and size. The process used to localize the target, once it is detected, is based on an inversion of the Pinhole camera model. The task of following the Summit XL was designed to keep the target inside the field of view of the camera, and it was implemented in the form of a PID controller. The system has been tested both in simulation and with real robots, showing promising results.

Mammalian capping enzyme binds RNA and uses protein tyrosine phosphatase mechanism

Mammalian capping enzymes are bifunctional proteins with both RNA 5′-triphosphatase and guanylyltransferase activities. The N-terminal 237-aa triphosphatase domain contains (I/V)HCXXGXXR(S/T)G, a sequence corresponding to the conserved active-site motif in protein tyrosine phosphatases (PTPs). Analysis of point mutants of mouse RNA 5′-triphosphatase identified the motif Cys and Arg residues and an upstream Asp as required for activity. Like PTPs, this enzyme was inhibited by iodoacetate and VO43− and independent of Mg2+, providing additional evidence for phosphate removal from RNA 5′ ends by a PTP-like mechanism. The full-length, 597-aa mouse capping enzyme and the C-terminal guanylyltransferase fragment (residues 211–597), unlike the triphosphatase domain, bound poly (U) and were nuclear in transfected cells. RNA binding was increased by GTP, and a guanylylation-defective, active-site mutant was not affected. Ala substitution at positions required for the formation of the enzyme-GMP capping intermediate (R315, R530, K533, or N537) also eliminated poly (U) binding, while proteins with conservative substitutions at these sites retained binding but not guanylyltransferase activity. These results demonstrate that the guanylyltransferase domain of mammalian capping enzyme specifies nuclear localization and RNA binding. Association of capping enzyme with nascent transcripts may act in synergy with RNA polymerase II binding to ensure 5′ cap formation.

mRNA binding protein mrnp 41 localizes to both nucleus and cytoplasm

We have identified and molecularly characterized a human protein with a Mr of 40,880 Da. After UV irradiation of HeLa cells, this protein was cross-linked to poly(A)-containing mRNA and was therefore designated mrnp 41 (for mRNA binding protein of 41 kDa). Cell fractionation and immunoblotting showed mrnp 41 in both the cytoplasm and the nucleus and particularly in the nuclear envelope. Immunofluorescence microscopy localized mrnp 41 to distinct foci in the nucleoplasm, to the nuclear rim, and to meshwork-like structures throughout the cytoplasm. The cytoplasmic meshwork staining was disrupted by prior treatment of cells with the actin filament- or microtubule-disrupting drugs cytochalasin or nocodazole, respectively, suggesting association of mrnp 41 with the cytoskeleton. Double immunofluorescence with antibodies against mrnp 41 and the cytoplasmic poly(A) binding protein showed colocalization to the cytoplasmic meshwork. Immunogold electronmicroscopy confirmed mrnp 41’s cytoplasmic and nucleoplasmic localization and revealed a striking labeling of nuclear pore complexes. Together these data suggest that mrnp 41 may function in nuclear export of mRNPs and/or in cytoplasmic transport on, or attachment to, the cytoskeleton. Consistent with a role of mrnp 41 in nuclear export are previous reports that mutations in homologs of mrnp 41 in Schizosaccharomyces pombe, designated Rae1p, or in Saccharomyces cerevisiae, designated Gle2p, result in mRNA accumulation in the nucleus although it is presently not known whether these homologs are mRNA binding proteins as well.