Computational methods to create and analyze a digital gene expression atlas of embryo development from microscopy images

Abstract The creation of atlases, or digital models where information from different subjects can be combined, is a field of increasing interest in biomedical imaging. When a single image does not contain enough information to appropriately describe the organism under study, it is then necessary to acquire images of several individuals, each of them containing complementary data with respect to the rest of the components in the cohort. This approach allows creating digital prototypes, ranging from anatomical atlases of human patients and organs, obtained for instance from Magnetic Resonance Imaging, to gene expression cartographies of embryo development, typically achieved from Light Microscopy. Within such context, in this PhD Thesis we propose, develop and validate new dedicated image processing methodologies that, based on image registration techniques, bring information from multiple individuals into alignment within a single digital atlas model. We also elaborate a dedicated software visualization platform to explore the resulting wealth of multi-dimensional data and novel analysis algo-rithms to automatically mine the generated resource in search of bio¬logical insights. In particular, this work focuses on gene expression data from developing zebrafish embryos imaged at the cellular resolution level with Two-Photon Laser Scanning Microscopy. Disposing of quantitative measurements relating multiple gene expressions to cell position and their evolution in time is a fundamental prerequisite to understand embryogenesis multi-scale processes. However, the number of gene expressions that can be simultaneously stained in one acquisition is limited due to optical and labeling constraints. These limitations motivate the implementation of atlasing strategies that can recreate a virtual gene expression multiplex. The developed computational tools have been tested in two different scenarios. The first one is the early zebrafish embryogenesis where the resulting atlas constitutes a link between the phenotype and the genotype at the cellular level. The second one is the late zebrafish brain where the resulting atlas allows studies relating gene expression to brain regionalization and neurogenesis. The proposed computational frameworks have been adapted to the requirements of both scenarios, such as the integration of partial views of the embryo into a whole embryo model with cellular resolution or the registration of anatom¬ical traits with deformable transformation models non-dependent on any specific labeling. The software implementation of the atlas generation tool (Match-IT) and the visualization platform (Atlas-IT) together with the gene expression atlas resources developed in this Thesis are to be made freely available to the scientific community. Lastly, a novel proof-of-concept experiment integrates for the first time 3D gene expression atlas resources with cell lineages extracted from live embryos, opening up the door to correlate genetic and cellular spatio-temporal dynamics. La creación de atlas, o modelos digitales, donde la información de distintos sujetos puede ser combinada, es un campo de creciente interés en imagen biomédica. Cuando una sola imagen no contiene suficientes datos como para describir apropiadamente el organismo objeto de estudio, se hace necesario adquirir imágenes de varios individuos, cada una de las cuales contiene información complementaria respecto al resto de componentes del grupo. De este modo, es posible crear prototipos digitales, que pueden ir desde atlas anatómicos de órganos y pacientes humanos, adquiridos por ejemplo mediante Resonancia Magnética, hasta cartografías de la expresión genética del desarrollo de embrionario, típicamente adquiridas mediante Microscopía Optica. Dentro de este contexto, en esta Tesis Doctoral se introducen, desarrollan y validan nuevos métodos de procesado de imagen que, basándose en técnicas de registro de imagen, son capaces de alinear imágenes y datos provenientes de múltiples individuos en un solo atlas digital. Además, se ha elaborado una plataforma de visualization específicamente diseñada para explorar la gran cantidad de datos, caracterizados por su multi-dimensionalidad, que resulta de estos métodos. Asimismo, se han propuesto novedosos algoritmos de análisis y minería de datos que permiten inspeccionar automáticamente los atlas generados en busca de conclusiones biológicas significativas. En particular, este trabajo se centra en datos de expresión genética del desarrollo embrionario del pez cebra, adquiridos mediante Microscopía dos fotones con resolución celular. Disponer de medidas cuantitativas que relacionen estas expresiones genéticas con las posiciones celulares y su evolución en el tiempo es un prerrequisito fundamental para comprender los procesos multi-escala característicos de la morfogénesis. Sin embargo, el número de expresiones genéticos que pueden ser simultáneamente etiquetados en una sola adquisición es reducido debido a limitaciones tanto ópticas como del etiquetado. Estas limitaciones requieren la implementación de estrategias de creación de atlas que puedan recrear un multiplexado virtual de expresiones genéticas. Las herramientas computacionales desarrolladas han sido validadas en dos escenarios distintos. El primer escenario es el desarrollo embrionario temprano del pez cebra, donde el atlas resultante permite constituir un vínculo, a nivel celular, entre el fenotipo y el genotipo de este organismo modelo. El segundo escenario corresponde a estadios tardíos del desarrollo del cerebro del pez cebra, donde el atlas resultante permite relacionar expresiones genéticas con la regionalización del cerebro y la formación de neuronas. La plataforma computacional desarrollada ha sido adaptada a los requisitos y retos planteados en ambos escenarios, como la integración, a resolución celular, de vistas parciales dentro de un modelo consistente en un embrión completo, o el alineamiento entre estructuras de referencia anatómica equivalentes, logrado mediante el uso de modelos de transformación deformables que no requieren ningún marcador específico. Está previsto poner a disposición de la comunidad científica tanto la herramienta de generación de atlas (Match-IT), como su plataforma de visualización (Atlas-IT), así como las bases de datos de expresión genética creadas a partir de estas herramientas. Por último, dentro de la presente Tesis Doctoral, se ha incluido una prueba conceptual innovadora que permite integrar los mencionados atlas de expresión genética tridimensionales dentro del linaje celular extraído de una adquisición in vivo de un embrión. Esta prueba conceptual abre la puerta a la posibilidad de correlar, por primera vez, las dinámicas espacio-temporales de genes y células.

Incorporating small-scale farmers into sustainable supply chains: the Manobi case

Increased globalization and outsourcing to developing countries is fostering the interest in supply chain sustainability. From the academic point of view, while environmental impacts of supply chains have been largely analysed, the research on social issues has been scattered and fragmented. This paper thereby sets out to close this gap. We have identified an emerging sphere of knowledge at the interface between sustainable supply chain management, business strategy and international development literature, which seeks to propose innovative strategies for poverty alleviation. The incorporation of impoverished farmers into supply chains is presented here as one of those strategies, and illustrated through a case study on the integration of these farmers in the Senegalese horticulture supply chain.

Qualitative operational risk software -development of a software tool for determining the root of operational risk

This article shows software that allows determining the statistical behavior of qualitative data originating surveys previously transformed with a Likert’s scale to quantitative data. The main intention is offer to users a useful tool to know statistics' characteristics and forecasts of financial risks in a fast and simple way. Additionally,this paper presents the definition of operational risk. On the other hand, the article explains different techniques to do surveys with a Likert’s scale (Avila, 2008) to know expert’s opinion with the transformation of qualitative data to quantitative data. In addition, this paper will show how is very easy to distinguish an expert’s opinion related to risk, but when users have a lot of surveys and matrices is very difficult to obtain results because is necessary to compare common data. On the other hand, statistical value representative must be extracted from common data to get weight of each risk. In the end, this article exposes the development of “Qualitative Operational Risk Software” or QORS by its acronym, which has been designed to determine the root of risks in organizations and its value at operational risk OpVaR (Jorion, 2008; Chernobai et al, 2008) when input data comes from expert’s opinion and their associated matrices.

Ventilation system design and large scale fire tests

The design, construction and operation of the tunnels of M-30, the major ring road in the city of Madrid (Spain), represent a very interesting project in wich a wide variety of situations -geometrical, topographical, etc.- had to be covered, in variable conditions of traffic. For that reasons, the M-30 project is a remarkable technical challenge, which, after its completion, turned into an international reference. From the "design for safety" perspective, a holistic approach has been used to deal with new technologies, integration of systems and development of the procedures to reach the maximum level. However, one of the primary goals has been to achieve reasonable homogeneity characteristics which can permit operate a netword of tunels as one only infraestructure. In the case of the ventilation system the mentioned goals have implied innovative solutions and coordination efforts of great interest. Consequently, this paper describes the principal ideas underlying the conceptual solution developed focusing on the principal peculiarities of the project.

Development of a methodology and tool to evaluate the impact of ICT measures on road transport emissions

The paper presents the main elements of a project entitled ICT-Emissions that aims at developing a novel methodology to evaluate the impact of ICT-related measures on mobility, vehicle energy consumption and CO2 emissions of vehicle fleets at the local scale, in order to promote the wider application of the most appropriate ICT measures. The proposed methodology combines traffic and emission modelling at micro and macro scales. These will be linked with interfaces and submodules which will be specifically designed and developed. A number of sources are available to the consortium to obtain the necessary input data. Also, experimental campaigns are offered to fill in gaps of information in traffic and emission patterns. The application of the methodology will be demonstrated using commercially available software. However, the methodology is developed in such a way as to enable its implementation by a variety of emission and traffic models. Particular emphasis is given to (a) the correct estimation of driver behaviour, as a result of traffic-related ICT measures, (b) the coverage of a large number of current vehicle technologies, including ICT systems, and (c) near future technologies such as hybrid, plug-in hybrids, and electric vehicles. The innovative combination of traffic, driver, and emission models produces a versatile toolbox that can simulate the impact on energy and CO2 of infrastructure measures (traffic management, dynamic traffic signs, etc.), driver assistance systems and ecosolutions (speed/cruise control, start/stop systems, etc.) or a combination of measures (cooperative systems).The methodology is validated by application in the Turin area and its capacity is further demonstrated by application in real world conditions in Madrid and Rome.

Estimation of nitrogen budgets for contrasting catchments at the landscape scale.

A comprehensive assessment of nitrogen (N) flows at the landscape scale is fundamental to understand spatial interactions in the N cascade and to inform the development of locally optimised N management strategies. To explore these interactions, complete N budgets were estimated for two contrasting hydrological catchments (dominated by agricultural grassland vs. semi-natural peat-dominated moorland), forming part of an intensively studied landscape in southern Scotland. Local scale atmospheric dispersion modelling and detailed farm and field inventories provided high resolution estimations of input fluxes. Direct agricultural inputs (i.e. grazing excreta, N2 fixation, organic and synthetic fertiliser) accounted for most of the catchment N inputs, representing 82% in the grassland and 62% in the moorland catchment, while atmospheric deposition made a significant contribution, particularly in the moorland catchment, contributing 38% of the N inputs. The estimated catchment N budgets highlighted areas of key uncertainty, particularly N2 exchange and stream N export. The resulting N balances suggest that the study catchments have a limited capacity to store N within soils, vegetation and groundwater. The "catchment N retention", i.e. the amount of N which is either stored within the catchment or lost through atmospheric emissions, was estimated to be 13% of the net anthropogenic input in the moorland and 61% in the grassland catchment. These values contrast with regional scale estimates: Catchment retentions of net anthropogenic input estimated within Europe at the regional scale range from 50% to 90%, with an average of 82% (Billen et al., 2011). This study emphasises the need for detailed budget analyses to identify the N status of European landscapes.

MIA - a free and open source software for gray scale medical image analysis

Background Gray scale images make the bulk of data in bio-medical image analysis, and hence, the main focus of many image processing tasks lies in the processing of these monochrome images. With ever improving acquisition devices, spatial and temporal image resolution increases, and data sets become very large. Various image processing frameworks exists that make the development of new algorithms easy by using high level programming languages or visual programming. These frameworks are also accessable to researchers that have no background or little in software development because they take care of otherwise complex tasks. Specifically, the management of working memory is taken care of automatically, usually at the price of requiring more it. As a result, processing large data sets with these tools becomes increasingly difficult on work station class computers. One alternative to using these high level processing tools is the development of new algorithms in a languages like C++, that gives the developer full control over how memory is handled, but the resulting workflow for the prototyping of new algorithms is rather time intensive, and also not appropriate for a researcher with little or no knowledge in software development. Another alternative is in using command line tools that run image processing tasks, use the hard disk to store intermediate results, and provide automation by using shell scripts. Although not as convenient as, e.g. visual programming, this approach is still accessable to researchers without a background in computer science. However, only few tools exist that provide this kind of processing interface, they are usually quite task specific, and don’t provide an clear approach when one wants to shape a new command line tool from a prototype shell script. Results The proposed framework, MIA, provides a combination of command line tools, plug-ins, and libraries that make it possible to run image processing tasks interactively in a command shell and to prototype by using the according shell scripting language. Since the hard disk becomes the temporal storage memory management is usually a non-issue in the prototyping phase. By using string-based descriptions for filters, optimizers, and the likes, the transition from shell scripts to full fledged programs implemented in C++ is also made easy. In addition, its design based on atomic plug-ins and single tasks command line tools makes it easy to extend MIA, usually without the requirement to touch or recompile existing code. Conclusion In this article, we describe the general design of MIA, a general purpouse framework for gray scale image processing. We demonstrated the applicability of the software with example applications from three different research scenarios, namely motion compensation in myocardial perfusion imaging, the processing of high resolution image data that arises in virtual anthropology, and retrospective analysis of treatment outcome in orthognathic surgery. With MIA prototyping algorithms by using shell scripts that combine small, single-task command line tools is a viable alternative to the use of high level languages, an approach that is especially useful when large data sets need to be processed.

New steps in the development of the second generation TEC GESMEY

The paper shows the results of the new steps that have been done in the development of the tidal energy converter GESMEY. These are the design, construction and trials into the sea of a 1/10 scale prototype and also the construction with the same scale of the buoy BOSCEM, that anchors the device and lets it in the correct work position and depth, along the two directions of the flow that the daily tidal cycle have. Inside the paper is described the objectives and the methodology of the experimental trials that were ca rry out the last summer with the scale prototype. GESMEY is a new type of tidal energy converter (TEC) that has the capability to exploit currents in waters over forty meters by itself and it gets only using its internal ballast system the necessary equilibrium between hy drostatics and hydrodynamics forces to make the emersion and the immersion procedures without any other help. Finally the paper shows the description of the results obtained over the performance of the devices along the immersion, emersion and floating transport manoeuvres and afterwards the results, that were obtained along the generation power tests that were carried out, are shown.

Smart aeronautical structures: development and experimental validation of a structural health monitoring system for damage detection

En muchas áreas de la ingeniería, la integridad y confiabilidad de las estructuras son aspectos de extrema importancia. Estos son controlados mediante el adecuado conocimiento de danos existentes. Típicamente, alcanzar el nivel de conocimiento necesario que permita caracterizar la integridad estructural implica el uso de técnicas de ensayos no destructivos. Estas técnicas son a menudo costosas y consumen mucho tiempo. En la actualidad, muchas industrias buscan incrementar la confiabilidad de las estructuras que emplean. Mediante el uso de técnicas de última tecnología es posible monitorizar las estructuras y en algunos casos, es factible detectar daños incipientes que pueden desencadenar en fallos catastróficos. Desafortunadamente, a medida que la complejidad de las estructuras, los componentes y sistemas incrementa, el riesgo de la aparición de daños y fallas también incrementa. Al mismo tiempo, la detección de dichas fallas y defectos se torna más compleja. En años recientes, la industria aeroespacial ha realizado grandes esfuerzos para integrar los sensores dentro de las estructuras, además de desarrollar algoritmos que permitan determinar la integridad estructural en tiempo real. Esta filosofía ha sido llamada “Structural Health Monitoring” (o “Monitorización de Salud Estructural” en español) y este tipo de estructuras han recibido el nombre de “Smart Structures” (o “Estructuras Inteligentes” en español). Este nuevo tipo de estructuras integran materiales, sensores, actuadores y algoritmos para detectar, cuantificar y localizar daños dentro de ellas mismas. Una novedosa metodología para detección de daños en estructuras se propone en este trabajo. La metodología está basada en mediciones de deformación y consiste en desarrollar técnicas de reconocimiento de patrones en el campo de deformaciones. Estas últimas, basadas en PCA (Análisis de Componentes Principales) y otras técnicas de reducción dimensional. Se propone el uso de Redes de difracción de Bragg y medidas distribuidas como sensores de deformación. La metodología se validó mediante pruebas a escala de laboratorio y pruebas a escala real con estructuras complejas. Los efectos de las condiciones de carga variables fueron estudiados y diversos experimentos fueron realizados para condiciones de carga estáticas y dinámicas, demostrando que la metodología es robusta ante condiciones de carga desconocidas. ABSTRACT In many engineering fields, the integrity and reliability of the structures are extremely important aspects. They are controlled by the adequate knowledge of existing damages. Typically, achieving the level of knowledge necessary to characterize the structural integrity involves the usage of nondestructive testing techniques. These are often expensive and time consuming. Nowadays, many industries look to increase the reliability of the structures used. By using leading edge techniques it is possible to monitoring these structures and in some cases, detect incipient damage that could trigger catastrophic failures. Unfortunately, as the complexity of the structures, components and systems increases, the risk of damages and failures also increases. At the same time, the detection of such failures and defects becomes more difficult. In recent years, the aerospace industry has done great efforts to integrate the sensors within the structures and, to develop algorithms for determining the structural integrity in real time. The ‘philosophy’ has being called “Structural Health Monitoring” and these structures have been called “smart structures”. These new types of structures integrate materials, sensors, actuators and algorithms to detect, quantify and locate damage within itself. A novel methodology for damage detection in structures is proposed. The methodology is based on strain measurements and consists in the development of strain field pattern recognition techniques. The aforementioned are based on PCA (Principal Component Analysis) and other dimensional reduction techniques. The use of fiber Bragg gratings and distributed sensing as strain sensors is proposed. The methodology have been validated by using laboratory scale tests and real scale tests with complex structures. The effects of the variable load conditions were studied and several experiments were performed for static and dynamic load conditions, demonstrating that the methodology is robust under unknown load conditions.

Technological development in Spanish gothic vaults design

The principle of complexity as the evolution vector of the gothic style was an idea largely developed by Paul Frankl. The high complexity reached in the 15th and 16th centuries was possible thanks to the geometrical resources developed in the workshops of the medieval stonemasons. The search for more sophisticated designs was possible also with the higher standardization, so that the most complex ribbed vault could be built with ribs that had all the same curvature and with voussoirs that were therefore identical. Spanish Gothic architecture has been deeply studied from a historical and artistic point of view. The present paper, as a complement to these analyses, aims to point out some of the geometrical methods and technological improvements that late medieval masons were able to develop. In that way, some selected vaults have been measured, in order to study their geometry and design process. Also scale models of some vaults have been built at the Escuela de Arquitectura (Madrid) to validate these geometrical principles. More than just a research method, the scale models allow to understand the medieval construction techniques, and they are a powerful pedagogical tool with which pupils can reach a rewarding experience based on the “medieval-way” praxis.

Development of a methodology for nuclear data uncertainty propagation on isotopic evolution calculations for advanced nuclear systems

Una apropiada evaluación de los márgenes de seguridad de una instalación nuclear, por ejemplo, una central nuclear, tiene en cuenta todas las incertidumbres que afectan a los cálculos de diseño, funcionanmiento y respuesta ante accidentes de dicha instalación. Una fuente de incertidumbre son los datos nucleares, que afectan a los cálculos neutrónicos, de quemado de combustible o activación de materiales. Estos cálculos permiten la evaluación de las funciones respuesta esenciales para el funcionamiento correcto durante operación, y también durante accidente. Ejemplos de esas respuestas son el factor de multiplicación neutrónica o el calor residual después del disparo del reactor. Por tanto, es necesario evaluar el impacto de dichas incertidumbres en estos cálculos. Para poder realizar los cálculos de propagación de incertidumbres, es necesario implementar metodologías que sean capaces de evaluar el impacto de las incertidumbres de estos datos nucleares. Pero también es necesario conocer los datos de incertidumbres disponibles para ser capaces de manejarlos. Actualmente, se están invirtiendo grandes esfuerzos en mejorar la capacidad de analizar, manejar y producir datos de incertidumbres, en especial para isótopos importantes en reactores avanzados. A su vez, nuevos programas/códigos están siendo desarrollados e implementados para poder usar dichos datos y analizar su impacto. Todos estos puntos son parte de los objetivos del proyecto europeo ANDES, el cual ha dado el marco de trabajo para el desarrollo de esta tesis doctoral. Por tanto, primero se ha llevado a cabo una revisión del estado del arte de los datos nucleares y sus incertidumbres, centrándose en los tres tipos de datos: de decaimiento, de rendimientos de fisión y de secciones eficaces. A su vez, se ha realizado una revisión del estado del arte de las metodologías para la propagación de incertidumbre de estos datos nucleares. Dentro del Departamento de Ingeniería Nuclear (DIN) se propuso una metodología para la propagación de incertidumbres en cálculos de evolución isotópica, el Método Híbrido. Esta metodología se ha tomado como punto de partida para esta tesis, implementando y desarrollando dicha metodología, así como extendiendo sus capacidades. Se han analizado sus ventajas, inconvenientes y limitaciones. El Método Híbrido se utiliza en conjunto con el código de evolución isotópica ACAB, y se basa en el muestreo por Monte Carlo de los datos nucleares con incertidumbre. En esta metodología, se presentan diferentes aproximaciones según la estructura de grupos de energía de las secciones eficaces: en un grupo, en un grupo con muestreo correlacionado y en multigrupos. Se han desarrollado diferentes secuencias para usar distintas librerías de datos nucleares almacenadas en diferentes formatos: ENDF-6 (para las librerías evaluadas), COVERX (para las librerías en multigrupos de SCALE) y EAF (para las librerías de activación). Gracias a la revisión del estado del arte de los datos nucleares de los rendimientos de fisión se ha identificado la falta de una información sobre sus incertidumbres, en concreto, de matrices de covarianza completas. Además, visto el renovado interés por parte de la comunidad internacional, a través del grupo de trabajo internacional de cooperación para evaluación de datos nucleares (WPEC) dedicado a la evaluación de las necesidades de mejora de datos nucleares mediante el subgrupo 37 (SG37), se ha llevado a cabo una revisión de las metodologías para generar datos de covarianza. Se ha seleccionando la actualización Bayesiana/GLS para su implementación, y de esta forma, dar una respuesta a dicha falta de matrices completas para rendimientos de fisión. Una vez que el Método Híbrido ha sido implementado, desarrollado y extendido, junto con la capacidad de generar matrices de covarianza completas para los rendimientos de fisión, se han estudiado diferentes aplicaciones nucleares. Primero, se estudia el calor residual tras un pulso de fisión, debido a su importancia para cualquier evento después de la parada/disparo del reactor. Además, se trata de un ejercicio claro para ver la importancia de las incertidumbres de datos de decaimiento y de rendimientos de fisión junto con las nuevas matrices completas de covarianza. Se han estudiado dos ciclos de combustible de reactores avanzados: el de la instalación europea para transmutación industrial (EFIT) y el del reactor rápido de sodio europeo (ESFR), en los cuales se han analizado el impacto de las incertidumbres de los datos nucleares en la composición isotópica, calor residual y radiotoxicidad. Se han utilizado diferentes librerías de datos nucleares en los estudios antreriores, comparando de esta forma el impacto de sus incertidumbres. A su vez, mediante dichos estudios, se han comparando las distintas aproximaciones del Método Híbrido y otras metodologías para la porpagación de incertidumbres de datos nucleares: Total Monte Carlo (TMC), desarrollada en NRG por A.J. Koning y D. Rochman, y NUDUNA, desarrollada en AREVA GmbH por O. Buss y A. Hoefer. Estas comparaciones demostrarán las ventajas del Método Híbrido, además de revelar sus limitaciones y su rango de aplicación. ABSTRACT For an adequate assessment of safety margins of nuclear facilities, e.g. nuclear power plants, it is necessary to consider all possible uncertainties that affect their design, performance and possible accidents. Nuclear data are a source of uncertainty that are involved in neutronics, fuel depletion and activation calculations. These calculations can predict critical response functions during operation and in the event of accident, such as decay heat and neutron multiplication factor. Thus, the impact of nuclear data uncertainties on these response functions needs to be addressed for a proper evaluation of the safety margins. Methodologies for performing uncertainty propagation calculations need to be implemented in order to analyse the impact of nuclear data uncertainties. Nevertheless, it is necessary to understand the current status of nuclear data and their uncertainties, in order to be able to handle this type of data. Great eórts are underway to enhance the European capability to analyse/process/produce covariance data, especially for isotopes which are of importance for advanced reactors. At the same time, new methodologies/codes are being developed and implemented for using and evaluating the impact of uncertainty data. These were the objectives of the European ANDES (Accurate Nuclear Data for nuclear Energy Sustainability) project, which provided a framework for the development of this PhD Thesis. Accordingly, first a review of the state-of-the-art of nuclear data and their uncertainties is conducted, focusing on the three kinds of data: decay, fission yields and cross sections. A review of the current methodologies for propagating nuclear data uncertainties is also performed. The Nuclear Engineering Department of UPM has proposed a methodology for propagating uncertainties in depletion calculations, the Hybrid Method, which has been taken as the starting point of this thesis. This methodology has been implemented, developed and extended, and its advantages, drawbacks and limitations have been analysed. It is used in conjunction with the ACAB depletion code, and is based on Monte Carlo sampling of variables with uncertainties. Different approaches are presented depending on cross section energy-structure: one-group, one-group with correlated sampling and multi-group. Differences and applicability criteria are presented. Sequences have been developed for using different nuclear data libraries in different storing-formats: ENDF-6 (for evaluated libraries) and COVERX (for multi-group libraries of SCALE), as well as EAF format (for activation libraries). A revision of the state-of-the-art of fission yield data shows inconsistencies in uncertainty data, specifically with regard to complete covariance matrices. Furthermore, the international community has expressed a renewed interest in the issue through the Working Party on International Nuclear Data Evaluation Co-operation (WPEC) with the Subgroup (SG37), which is dedicated to assessing the need to have complete nuclear data. This gives rise to this review of the state-of-the-art of methodologies for generating covariance data for fission yields. Bayesian/generalised least square (GLS) updating sequence has been selected and implemented to answer to this need. Once the Hybrid Method has been implemented, developed and extended, along with fission yield covariance generation capability, different applications are studied. The Fission Pulse Decay Heat problem is tackled first because of its importance during events after shutdown and because it is a clean exercise for showing the impact and importance of decay and fission yield data uncertainties in conjunction with the new covariance data. Two fuel cycles of advanced reactors are studied: the European Facility for Industrial Transmutation (EFIT) and the European Sodium Fast Reactor (ESFR), and response function uncertainties such as isotopic composition, decay heat and radiotoxicity are addressed. Different nuclear data libraries are used and compared. These applications serve as frameworks for comparing the different approaches of the Hybrid Method, and also for comparing with other methodologies: Total Monte Carlo (TMC), developed at NRG by A.J. Koning and D. Rochman, and NUDUNA, developed at AREVA GmbH by O. Buss and A. Hoefer. These comparisons reveal the advantages, limitations and the range of application of the Hybrid Method.

Lidar uncertainty in complex terrain development of a bias correction methodology

El autor ha trabajado como parte del equipo de investigación en mediciones de viento en el Centro Nacional de Energías Renovables (CENER), España, en cooperación con la Universidad Politécnica de Madrid y la Universidad Técnica de Dinamarca. El presente reporte recapitula el trabajo de investigación realizado durante los últimos 4.5 años en el estudio de las fuentes de error de los sistemas de medición remota de viento, basados en la tecnología lidar, enfocado al error causado por los efectos del terreno complejo. Este trabajo corresponde a una tarea del paquete de trabajo dedicado al estudio de sistemas remotos de medición de viento, perteneciente al proyecto de intestigación europeo del 7mo programa marco WAUDIT. Adicionalmente, los datos de viento reales han sido obtenidos durante las campañas de medición en terreno llano y terreno complejo, pertenecientes al también proyecto de intestigación europeo del 7mo programa marco SAFEWIND. El principal objetivo de este trabajo de investigación es determinar los efectos del terreno complejo en el error de medición de la velocidad del viento obtenida con los sistemas de medición remota lidar. Con este conocimiento, es posible proponer una metodología de corrección del error de las mediciones del lidar. Esta metodología está basada en la estimación de las variaciones del campo de viento no uniforme dentro del volumen de medición del lidar. Las variaciones promedio del campo de viento son predichas a partir de los resultados de las simulaciones computacionales de viento RANS, realizadas para el parque experimental de Alaiz. La metodología de corrección es verificada con los resultados de las simulaciones RANS y validadas con las mediciones reales adquiridas en la campaña de medición en terreno complejo. Al inicio de este reporte, el marco teórico describiendo el principio de medición de la tecnología lidar utilizada, es presentado con el fin de familiarizar al lector con los principales conceptos a utilizar a lo largo de este trabajo. Posteriormente, el estado del arte es presentado en donde se describe los avances realizados en el desarrollo de la la tecnología lidar aplicados al sector de la energía eólica. En la parte experimental de este trabajo de investigación se ha estudiado los datos adquiridos durante las dos campañas de medición realizadas. Estas campañas has sido realizadas en terreno llano y complejo, con el fin de complementar los conocimiento adquiridos en casa una de ellas y poder comparar los efectos del terreno en las mediciones de viento realizadas con sistemas remotos lidar. La primer campaña experimental se desarrollo en terreno llano, en el parque de ensayos de aerogeneradores H0vs0re, propiedad de DTU Wind Energy (anteriormente Ris0). La segunda campaña experimental se llevó a cabo en el parque de ensayos de aerogeneradores Alaiz, propiedad de CENER. Exactamente los mismos dos equipos lidar fueron utilizados en estas campañas, haciendo de estos experimentos altamente relevantes en el contexto de evaluación del recurso eólico. Un equipo lidar está basado en tecnología de onda continua, mientras que el otro está basado en tecnología de onda pulsada. La velocidad del viento fue medida, además de con los equipos lidar, con anemómetros de cazoletas, veletas y anemómetros verticales, instalados en mástiles meteorológicos. Los sensores del mástil meteorológico son considerados como las mediciones de referencia en el presente estudio. En primera instancia, se han analizado los promedios diez minútales de las medidas de viento. El objetivo es identificar las principales fuentes de error en las mediciones de los equipos lidar causadas por diferentes condiciones atmosféricas y por el flujo no uniforme de viento causado por el terreno complejo. El error del lidar ha sido estudiado como función de varias propiedades estadísticas del viento, como lo son el ángulo vertical de inclinación, la intensidad de turbulencia, la velocidad vertical, la estabilidad atmosférica y las características del terreno. El propósito es usar este conocimiento con el fin de definir criterios de filtrado de datos. Seguidamente, se propone una metodología para corregir el error del lidar causado por el campo de viento no uniforme, producido por la presencia de terreno complejo. Esta metodología está basada en el análisis matemático inicial sobre el proceso de cálculo de la velocidad de viento por los equipos lidar de onda continua. La metodología de corrección propuesta hace uso de las variaciones de viento calculadas a partir de las simulaciones RANS realizadas para el parque experimental de Alaiz. Una ventaja importante que presenta esta metodología es que las propiedades el campo de viento real, presentes en las mediciones instantáneas del lidar de onda continua, puede dar paso a análisis adicionales como parte del trabajo a futuro. Dentro del marco del proyecto, el trabajo diario se realizó en las instalaciones de CENER, con supervisión cercana de la UPM, incluyendo una estancia de 1.5 meses en la universidad. Durante esta estancia, se definió el análisis matemático de las mediciones de viento realizadas por el equipo lidar de onda continua. Adicionalmente, los efectos del campo de viento no uniforme sobre el error de medición del lidar fueron analíticamente definidos, después de asumir algunas simplificaciones. Adicionalmente, durante la etapa inicial de este proyecto se desarrollo una importante trabajo de cooperación con DTU Wind Energy. Gracias a esto, el autor realizó una estancia de 1.5 meses en Dinamarca. Durante esta estancia, el autor realizó una visita a la campaña de medición en terreno llano con el fin de aprender los aspectos básicos del diseño de campañas de medidas experimentales, el estudio del terreno y los alrededores y familiarizarse con la instrumentación del mástil meteorológico, el sistema de adquisición y almacenamiento de datos, así como de el estudio y reporte del análisis de mediciones. ABSTRACT The present report summarizes the research work performed during last 4.5 years of investigation on the sources of lidar bias due to complex terrain. This work corresponds to one task of the remote sensing work package, belonging to the FP7 WAUDIT project. Furthermore, the field data from the wind velocity measurement campaigns of the FP7 SafeWind project have been used in this report. The main objective of this research work is to determine the terrain effects on the lidar bias in the measured wind velocity. With this knowledge, it is possible to propose a lidar bias correction methodology. This methodology is based on an estimation of the wind field variations within the lidar scan volume. The wind field variations are calculated from RANS simulations performed from the Alaiz test site. The methodology is validated against real scale measurements recorded during an eight month measurement campaign at the Alaiz test site. Firstly, the mathematical framework of the lidar sensing principle is introduced and an overview of the state of the art is presented. The experimental part includes the study of two different, but complementary experiments. The first experiment was a measurement campaign performed in flat terrain, at DTU Wind Energy H0vs0re test site, while the second experiment was performed in complex terrain at CENER Alaiz test site. Exactly the same two lidar devices, based on continuous wave and pulsed wave systems, have been used in the two consecutive measurement campaigns, making this a relevant experiment in the context of wind resource assessment. The wind velocity was sensed by the lidars and standard cup anemometry and wind vanes (installed on a met mast). The met mast sensors are considered as the reference wind velocity measurements. The first analysis of the experimental data is dedicated to identify the main sources of lidar bias present in the 10 minute average values. The purpose is to identify the bias magnitude introduced by different atmospheric conditions and by the non-uniform wind flow resultant of the terrain irregularities. The lidar bias as function of several statistical properties of the wind flow like the tilt angle, turbulence intensity, vertical velocity, atmospheric stability and the terrain characteristics have been studied. The aim of this exercise is to use this knowledge in order to define useful lidar bias data filters. Then, a methodology to correct the lidar bias caused by non-uniform wind flow is proposed, based on the initial mathematical analysis of the lidar measurements. The proposed lidar bias correction methodology has been developed focusing on the the continuous wave lidar system. In a last step, the proposed lidar bias correction methodology is validated with the data of the complex terrain measurement campaign. The methodology makes use of the wind field variations obtained from the RANS analysis. The results are presented and discussed. The advantage of this methodology is that the wind field properties at the Alaiz test site can be studied with more detail, based on the instantaneous measurements of the CW lidar. Within the project framework, the daily basis work has been done at CENER, with close guidance and support from the UPM, including an exchange period of 1.5 months. During this exchange period, the mathematical analysis of the lidar sensing of the wind velocity was defined. Furthermore, the effects of non-uniform wind fields on the lidar bias were analytically defined, after making some assumptions for the sake of simplification. Moreover, there has been an important cooperation with DTU Wind Energy, where a secondment period of 1.5 months has been done as well. During the secondment period at DTU Wind Energy, an important introductory learning has taken place. The learned aspects include the design of an experimental measurement campaign in flat terrain, the site assessment study of obstacles and terrain conditions, the data acquisition and processing, as well as the study and reporting of the measurement analysis.

Large-scale identification of gibberellin-related transcription factor defines group VII ERFs as functional of DELLA partners

DELLA proteins are the master negative regulators in gibberellin (GA) signaling acting in the nucleus as transcriptional regulators. The current view of DELLA action indicates that their activity relies on the physical interaction with transcription factors (TFs). Therefore, the identification of TFs through which DELLAs regulate GA responses is key to understanding these responses from a mechanistic point of view. Here, we have determined the TF interactome of the Arabidopsis (Arabidopsis thaliana) DELLA protein GIBBERELLIN INSENSITIVE and screened a collection of conditional TF overexpressors in search of those that alter GA sensitivity. As a result, we have found RELATED TO APETALA2.3, an ethylene-induced TF belonging to the group VII ETHYLENE RESPONSE FACTOR of the APETALA2/ethylene responsive element binding protein superfamily, as a DELLA interactor with physiological relevance in the context of apical hook development. The combination of transactivation assays and chromatin immunoprecipitation indicates that the interaction with GIBBERELLIN INSENSITIVE impairs the activity of RELATED TO APETALA2.3 on the target promoters. This mechanism represents a unique node in the cross regulation between the GA and ethylene signaling pathways controlling differential growth during apical hook development.

Development of novel melt-processable biopolymer nanocomposites based on poly(L-lactic acid) and WS2 inorganic nanotubes.

The use of tungsten disulphide inorganic nanotubes (INT-WS2) offers the opportunity to produce novel and advanced biopolymer-based nanocomposite materials with excellent nanoparticle dispersion without the need for modifiers or surfactants via conventional melt blending. The study of the non-isothermal melt-crystallization kinetics provides a clear picture of the transformation of poly(L-lactic acid) (PLLA) molecules from the non-ordered to the ordered state. The overall crystallization rate, final crystallinity and subsequent melting behaviour of PLLA were controlled by both the incorporation of INT-WS2 and the variation of the cooling rate. In particular, it was shown that INT-WS2 exhibits much more prominent nucleation activity on the crystallization of PLLA than other specific nucleating agents or nano-sized fillers. These features may be advantageous for the enhancement of mechanical properties and process-ability of PLLA-based materials. PLLA/INT-WS2 nanocomposites can be employed as low cost biodegradable materials for many eco-friendly and medical applications, and the exceptional crystallization behaviour observed opens new perspectives for scale-up and broader applications.

Guidelines for the design of an environmental flow regime program at the regional scale: Application of the ELOHA framework in the Ebro River basin

La gestión de los recursos hídricos se convierte en un reto del presente y del futuro frente a un panorama de continuo incremento de la demanda de agua debido al crecimiento de la población, el crecimiento del desarrollo económico y los posibles efectos del calentamiento global. La política hidráulica desde los años 60 en España se ha centrado en la construcción de infraestructuras que han producido graves alteraciones en el régimen natural de los ríos. Estas alteraciones han provocado y acrecentado los impactos sobre los ecosistemas fluviales y ribereños. Desde los años 90, sin embargo, ha aumentado el interés de la sociedad para conservar estos ecosistemas. El concepto de caudales ambientales consiste en un régimen de caudales que simula las características principales del régimen natural. Los caudales ambientales están diseñados para conservar la estructura y funcionalidad de los ecosistemas asociados al régimen fluvial, bajo la hipótesis de que los elementos que conforman estos ecosistemas están profundamente adaptados al régimen natural de caudales, y que cualquier alteración del régimen natural puede provocar graves daños a todo el sistema. El método ELOHA (Ecological Limits of Hydrological Alteration) tiene como finalidad identificar las componentes del régimen natural de caudales que son clave para mantener el equilibrio de los ecosistemas asociados, y estimar los límites máximos de alteración de estas componentes para garantizar su buen estado. Esta tesis presenta la aplicación del método ELOHA en la cuenca del Ebro. La cuenca del Ebro está profundamente regulada e intervenida por el hombre, y sólo las cabeceras de los principales afluentes del Ebro gozan todavía de un régimen total o cuasi natural. La tesis se estructura en seis capítulos que desarrollan las diferentes partes del método. El primer capítulo explica cómo se originó el concepto “caudales ambientales” y en qué consiste el método ELOHA. El segundo capítulo describe el área de estudio. El tercer capítulo realiza una clasificación de los regímenes naturales de la cuenca (RNC) del Ebro, basada en series de datos de caudal mínimamente alterado y usando exclusivamente parámetros hidrológicos. Se identificaron seis tipos diferentes de régimen natural: pluvial mediterráneo, nivo-pluvial, pluvial mediterréaneo con una fuerte componente del caudal base, pluvial oceánico, pluvio-nival oceánico y Mediterráneo. En el cuarto capítulo se realiza una regionalización a toda la cuenca del Ebro de los seis RNC encontrados en la cueca. Mediante parámetros climáticos y fisiográficos se extrapola la información del tipo de RNC a puntos donde no existen datos de caudal inalterado. El patrón geográfico de los tipos de régimen fluvial obtenido con la regionalización resultó ser coincidente con el patrón obtenido a través de la clasificación hidrológica. El quinto capítulo presenta la validación biológica de los procesos de clasificación anteriores: clasificación hidrológica y regionalización. La validación biológica de los tipos de regímenes fluviales es imprescindible, puesto que los diferentes tipos de régimen fluvial van a servir de unidades de gestión para favorecer el mantenimiento de los ecosistemas fluviales. Se encontraron diferencias significativas entre comunidades biológicas en cinco de los seis tipos de RNC encontrados en la cuenca. Finalmente, en el sexto capítulo se estudian las relaciones hidro-ecológicas existentes en tres de los seis tipos de régimen fluvial encontrados en la cuenca del Ebro. Mediante la construcción de curvas hidro-ecológicas a lo largo de un gradiente de alteración hidrológica, se pueden sugerir los límites de alteración hidrológica (ELOHAs) para garantizar el buen estado ecológico en cada uno de los tipos fluviales estudiados. Se establecieron ELOHAs en tres de los seis tipos de RNC de la cuenca del Ebro Esta tesis, además, pone en evidencia la falta de datos biológicos asociados a registros de caudal. Para llevar a cabo la implantación de un régimen de caudales ambientales en la cuenca, la ubicación de los puntos de muestreo biológico cercanos a estaciones de aforo es imprescindible para poder extraer relaciones causa-efecto de la gestión hidrológica sobre los ecosistemas dependientes. ABSTRACT In view of a growing freshwater demand because of population raising, improvement of economies and the potential effects of climate change, water resources management has become a challenge for present and future societies. Water policies in Spain have been focused from the 60’s on constructing hydraulic infrastructures, in order to dampen flow variability and granting water availability along the year. Consequently, natural flow regimes have been deeply altered and so the depending habitats and its ecosystems. However, an increasing acknowledgment of societies for preserving healthy freshwater ecosystems started in the 90’s and agreed that to maintain healthy freshwater ecosystems, it was necessary to set environmental flow regimes based on the natural flow variability. The Natural Flow Regime paradigm (Richter et al. 1996, Poff et al. 1997) bases on the hypothesis that freshwater ecosystems are made up by elements adapted to natural flow conditions, and any change on these conditions can provoke deep impacts on the whole system. Environmental flow regime concept consists in designing a flow regime that emulates natural flow characteristics, so that ecosystem structure, functions and services are maintained. ELOHA framework (Ecological Limits of Hydrological Alteration) aims to identify key features of the natural flow regime (NFR) that are needed to maintain and preserve healthy freshwater and riparian ecosystems. Moreover, ELOHA framework aims to quantify thresholds of alteration of these flow features according to ecological impacts. This thesis describes the application of the ELOHA framework in the Ebro River Basin. The Ebro River basin is the second largest basin in Spain and it is highly regulated for human demands. Only the Ebro headwaters tributaries still have completely unimpaired flow regime. The thesis has six chapters and the process is described step by step. The first chapter makes an introduction to the origin of the environmental flow concept and the necessity to come up. The second chapter shows a description of the study area. The third chapter develops a classification of NFRs in the basin based on natural flow data and using exclusively hydrological parameters. Six NFRs were found in the basin: continental Mediterranean-pluvial, nivo-pluvial, continental Mediterranean pluvial (with groundwater-dominated flow pattern), pluvio-oceanic, pluvio-nival-oceanic and Mediterranean. The fourth chapter develops a regionalization of the six NFR types across the basin by using climatic and physiographic variables. The geographical pattern obtained from the regionalization process was consistent with the pattern obtained with the hydrologic classification. The fifth chapter performs a biological validation of both classifications, obtained from the hydrologic classification and the posterior extrapolation. When the aim of flow classification is managing water resources according to ecosystem requirements, a validation based on biological data is compulsory. We found significant differences in reference macroinvertebrate communities between five over the six NFR types identified in the Ebro River basin. Finally, in the sixth chapter we explored the existence of significant and explicative flow alteration-ecological response relationships (FA-E curves) within NFR types in the Ebro River basin. The aim of these curves is to find out thresholds of hydrological alteration (ELOHAs), in order to preserve healthy freshwater ecosystem. We set ELOHA values in three NFR types identified in the Ebro River basin. During the development of this thesis, an inadequate biological monitoring in the Ebro River basin was identified. The design and establishment of appropriate monitoring arrangements is a critical final step in the assessment and implementation of environmental flows. Cause-effect relationships between hydrology and macroinvertebrate community condition are the principal data that sustain FA-E curves. Therefore, both data sites must be closely located, so that the effects of external factors are minimized. The scarce hydro-biological pairs of data available in the basin prevented us to apply the ELOHA method at all NFR types.