Aphids in Corn Research

Corn, Zea mays L., is the most abundant field crop in Iowa, and there are many insect pests associated with this field crop. Although aphids are not typically economically important in corn, recent observations have indicated several aphid species developing heavy populations in northwest Iowa and southwest Minnesota. Historically, the corn leaf aphid, Rhopalosiphum maidis (Hemiptera: Aphididae), has been the most abundant aphid species in corn; however, the bird cherry oat aphid, R. padi (Hemiptera: Aphididae), and several other species have also been detected. Recent observations show a shift to populations peaking later in the summer. Damage potential and management guidelines for aphids in corn are not well defined and this research is aimed at developing economic threshold and sampling protocols.

Data from PhD thesis

The ocean quahog, Arctica islandica is the longest-lived non-colonial animal known to science. A maximum individual age of this bivalve of 405 years has been found in a population off the north western coast of Iceland. Conspicuously shorter maximum lifespan potentials (MLSPs) were recorded from other populations of A. islandica in European waters (e.g. Kiel Bay: 30 years, German Bight: 150 years) which experience wider temperature and salinity fluctuations than the clams from Iceland. The aim of my thesis was to identify possible life-prolonging physiological strategies in A. islandica and to examine the modulating effects of extrinsic factors (e.g. seawater temperature, food availability) and intrinsic factors (e.g. species-specific behavior) on these strategies. Burrowing behavior and metabolic rate depression (MRD), tissue-specific antioxidant and anaerobic capacities as well as cell-turnover (= apoptosis and proliferation) rates were investigated in A. islandica from Iceland and the German Bight. An inter-species comparison of the quahog with the epibenthic scallop Aequipecten opercularis (MLSP = 8-10 years) was carried out in order to determine whether bivalves with short lifespans and different lifestyles also feature a different pattern in cellular maintenance and repair. The combined effects of a low-metabolic lifestyle, low oxidative damage accumulation, and constant investment into cellular protection and tissue maintenance, appear to slow-down the process of physiological aging in A. islandica and to afford the extraordinarily long MLSP in this species. Standard metabolic rates were lower in A. islandica when compared to the shorter-lived A. opercularis. Furthermore, A. islandica regulate mantle cavity water PO2 to mean values < 5 kPa, a PO2 at which the formation of reactive oxygen species (ROS) in isolated gill tissues of the clams was found to be 10 times lower than at normoxic conditions (21 kPa). Burrowing and metabolic rate depression (MRD) in Icelandic specimens were more pronounced in winter, possibly supported by low seawater temperature and food availability, and seem to be key energy-saving and life-prolonging parameters in A. islandica. The signaling molecule nitric oxide (NO) may play an important role during the onset of MRD in the ocean quahog by directly inhibiting cytochome-c-oxidase at low internal oxygenation upon shell closure. In laboratory experiments, respiration of isolated A. islandica gills was completely inhibited by chemically produced NO at low experimental PO2 <= 10 kPa. During shell closure, mantle cavity water PO2 decreased to 0 kPa for longer than 24 h, a state in which ROS production is supposed to subside. Compared to other mollusk species, onset of anaerobic metabolism is late in A. islandica in the metabolically reduced state. Increased accumulation of the anaerobic metabolite succinate was initially detected in the adductor muscle of the clams after 3.5 days under anoxic incubation or in burrowed specimens. A ROS-burst was absent in isolated gill tissue of the clams following hypoxia (5 kPa)-reoxygenation (21 kPa). Accordingly, neither the activity of antioxidant enzymes superoxide dismutase (SOD) and catalase (CAT), nor the specific content of the ROS-scavenger glutathione (GSH) was enhanced in different tissues of the ocean quahog after 3.5 days of self-induced or forced hypoxia/anoxia to prepare for an oxidative burst. While reduced ROS formation compared to routine levels lowers oxidative stress during MRD and also during surfacing, the general preservation of high cellular defense and the efficient removal and replacement of damaged cells over lifetime seem to be of crucial importance in decelerating the senescent decline in tissues of A. islandica. Along with stable antioxidant protection over 200 years of age, proliferation rates and apoptosis intensities in most investigated tissues of the ocean quahog were low, but constant over 140 years of age. Accordingly, age-dependent accumulations of protein and lipid oxidation products are lower in A. islandica tissues when compared to the shorter-lived bivalve A. opercularis. The short-lived swimming scallop is a model bivalve species representing the opposite life and aging strategy to A. islandica. In this species permanently high energy throughput, reduced investment into antioxidant defense with age, and higher accumulation of oxidation products are met by higher cell turnover rates than in the ocean quahog. The only symptoms of physiological change over age ever found in A. islandica were decreasing cell turnover rates in the heart muscle over a lifetime of 140 years. This may either indicate higher damage levels and possibly ongoing loss of functioning in the heart of aging clams, or, the opposite, lower rates of cell damage and a reduced need for cell renewal in the heart tissue of A. islandica over lifetime. Basic physiological capacities of different A. islandica populations, measured at controlled laboratory conditions, could not explain considerable discrepancies in population specific MLSPs. For example, levels of tissue-specific antioxidant capacities and cell turnover rates were similarly high in individuals from the German Bight and from Iceland. Rather than genetic differences, the local impacts of environmental conditions on behavioral and physiological traits in the ocean quahog seem to be responsible for differences in population-specific MLSPs.

Urban forest health survey on Roznik in Ljubljana in 2013 by two methods

Urban forest health was surveyed on Roznik in Ljubljana (46.05141 N, 14.47797 E) in 2013 by two methods: ICP Forests and UFMO. ICP Forests is most commonly used monitoring programme in Europe - the International Co-operative Programme on the Assessment and Monitoring of Air Pollution Effects on Forests, which is based on systematic grid. UFMO method - Urban Forests Management Oriented method was developed in the frame of EMoNFUr Project - Establishing a monitoring network to assess lowland forest and urban plantations in Lombardy and urban forest in Slovenia (LIFE10 ENV/IT/000399). UFMO is based on non-linear transects (GPS tracks). ICP forests monitoring plots were established in July 2013 in the urban forest Roznik in Ljubljana .The 32 plots are located on sampling grid 500 × 500 m. The grid was down-scaled from the National Forest Monitoring survey, which bases on national sample grid 4 × 4 km. With the ICP forests method the following parameters for each tree within the 15 plots were gathered according to the ICP forests manual for Visual assessment of crown condition and damaging agents: tree species, percentage of defoliation, affected part of the tree, specification of affected part, location in crown, symptom, symptom specification, causal agents / factors, age of damage, damage extent, and damage extent on the trunk. With the UFMO method, the following parameters for each tree that needed sylviculture measure (felling, pruning, sanitary felling, thinning, etc.) were recorded: tree species, breast diameter, causal agent / damaging factor, GPS waypoint and GPS track. For overall picture in the urban forest health problems, also other biotic and abiotic damaging factors that did not require management action were recorded.

Estudio de la tolerancia al daño de los materiales y sistemas de pretensado

La mayoría de las estructuras de hormigón pretensadas construidas en los últimos 50 años han demostrado una excelente durabilidad cuando su construcción se realiza atendiendo las recomendaciones de un buen diseño así como una buena ejecución y puesta en obra de la estructura. Este hecho se debe en gran parte al temor que despierta el fenómeno de la corrosión bajo tensión típico de las armaduras de acero de alta resistencia. Menos atención se ha prestado a la susceptibilidad a la corrosión bajo tensión de los anclajes de postensado, posiblemente debido a que se han reportado pocos casos de fallos catastróficos. El concepto de Tolerancia al Daño y la Mecánica de la Fractura en estructuras de Ingeniería Civil ha empezado a incorporarse recientemente en algunas normas de diseño y cálculo de estructuras metálicas, sin embargo, aún está lejos de ser asimilado y empleado habitualmente por los ingenieros en sus cálculos cuando la ocasión lo requiere. Este desconocimiento de los aspectos relacionados con la Tolerancia al Daño genera importantes gastos de mantenimiento y reparación. En este trabajo se ha estudiado la aplicabilidad de los conceptos de la Mecánica de la Fractura a los componentes de los sistemas de postensado empleados en ingeniería civil, empleándolo para analizar la susceptibilidad de las armaduras activas frente a la corrosión bajo tensiones y a la pérdida de capacidad portante de las cabezas de anclajes de postensado debido a la presencia de defectos. Con este objeto se han combinado tanto técnicas experimentales como numéricas. Los defectos superficiales en los alambres de pretensado no se presentan de manera aislada si no que existe una cierta continuidad en la dirección axial así como un elevado número de defectos. Por este motivo se ha optado por un enfoque estadístico, que es más apropiado que el determinístico. El empleo de modelos estadísticos basados en la teoría de valores extremos ha permitido caracterizar el estado superficial en alambres de 5,2 mm de diámetro. Por otro lado la susceptibilidad del alambre frente a la corrosión bajo tensión ha sido evaluada mediante la realización de una campaña de ensayos de acuerdo con la actual normativa que ha permitido caracterizar estadísticamente su comportamiento. A la vista de los resultados ha sido posible evaluar como los parámetros que definen el estado superficial del alambre pueden determinar la durabilidad de la armadura atendiendo a su resistencia frente a la corrosión bajo tensión, evaluada mediante los ensayos que especifica la normativa. En el caso de las cabezas de anclaje de tendones de pretensado, los defectos se presentan de manera aislada y tienen su origen en marcas, arañazos o picaduras de corrosión que pueden producirse durante el proceso de fabricación, transporte, manipulación o puesta en obra. Dada la naturaleza de los defectos, el enfoque determinístico es más apropiado que el estadístico. La evaluación de la importancia de un defecto en un elemento estructural requiere la estimación de la solicitación local que genera el defecto, que permite conocer si el defecto es crítico o si puede llegar a serlo, si es que progresa con el tiempo (por fatiga, corrosión, una combinación de ambas, etc.). En este trabajo los defectos han sido idealizados como grietas, de manera que el análisis quedara del lado de la seguridad. La evaluación de la solicitación local del defecto ha sido calculada mediante el empleo de modelos de elementos finitos de la cabeza de anclaje que simulan las condiciones de trabajo reales de la cabeza de anclaje durante su vida útil. A partir de estos modelos numéricos se ha analizado la influencia en la carga de rotura del anclaje de diversos factores como la geometría del anclaje, las condiciones del apoyo, el material del anclaje, el tamaño del defecto su forma y su posición. Los resultados del análisis numérico han sido contrastados satisfactoriamente mediante la realización de una campaña experimental de modelos a escala de cabezas de anclaje de Polimetil-metacrilato en los que artificialmente se han introducido defectos de diversos tamaños y en distintas posiciones. ABSTRACT Most of the prestressed concrete structures built in the last 50 years have demonstrated an excellent durability when they are constructed in accordance with the rules of good design, detailing and execution. This is particularly true with respect to the feared stress corrosion cracking, which is typical of high strength prestressing steel wires. Less attention, however, has been paid to the stress corrosion cracking susceptibility of anchorages for steel tendons for prestressing concrete, probably due to the low number of reported failure cases. Damage tolerance and fracture mechanics concepts in civil engineering structures have recently started to be incorporated in some design and calculation rules for metallic structures, however it is still far from being assimilated and used by civil engineers in their calculations on a regular basis. This limited knowledge of the damage tolerance basis could lead to significant repair and maintenance costs. This work deals with the applicability of fracture mechanics and damage tolerance concepts to the components of prestressed systems, which are used in civil engineering. Such concepts have been applied to assess the susceptibility of the prestressing steel wires to stress corrosion cracking and the reduction of load bearing capability of anchorage devices due to the presence of defects. For this purpose a combination of experimental work and numerical techniques have been performed. Surface defects in prestressing steel wires are not shown alone, though a certain degree of continuity in the axial direction exist. A significant number of such defects is also observed. Hence a statistical approach was used, which is assumed to be more appropriate than the deterministic approach. The use of statistical methods based in extreme value theories has allowed the characterising of the surface condition of 5.2 mm-diameter wires. On the other hand the stress corrosion cracking susceptibility of the wire has been assessed by means of an experimental testing program in line with the current regulations, which has allowed statistical characterisasion of their performances against stress corrosion cracking. In the light of the test results, it has been possible to evaluate how the surface condition parameters could determine the durability of the active metal armour regarding to its resistance against stress corrosion cracking assessed by means of the current testing regulations. In the case of anchorage devices for steel tendons for prestressing concrete, the damage is presented as point defects originating from dents, scratches or corrosion pits that could be produced during the manufacturing proccess, transport, handling, assembly or use. Due to the nature of these defects, in this case the deterministic approach is more appropriate than the statistical approach. The assessment of the relevancy of defect in a structural component requires the computation of the stress intensity factors, which in turn allow the evaluation of whether the size defect is critical or could become critical with the progress of time (due to fatigue, corrosion or a combination of both effects). In this work the damage is idealised as tiny cracks, a conservative hypothesis. The stress intensity factors have been calculated by means of finite element models of the anchorage representing the real working conditions during its service life. These numeric models were used to assess the impact of some factors on the rupture load of the anchorage, such the anchorage geometry, material, support conditions, defect size, shape and its location. The results from the numerical analysis have been succesfully correlated against the results of the experimental testing program of scaled models of the anchorages in poly-methil methacrylate in which artificial damage in several sizes and locations were introduced.

Detección de marcas para realidad aumentada sobre dispositivos móviles

Este documento es una guía para el desarrollo de una aplicación para dispositivos móviles en Android. Dicha aplicación combina las técnicas de visión por computador para calibrar la cámara del dispositivo y localizar un elemento en el espacio en base a esos los parámetros calculados en la calibración. El diseño de la aplicación incluye las decisiones sobre la forma en que se reciben los inputs de la aplicación, que patrones se utilizan en la calibración y en la localización y como se muestran los resultados finales al usuario. También incluye un diagrama de flujo de información que representa el tránsito de esta entre los diferentes módulos. La implementación comienza con la configuración de un entorno para desarrollar aplicaciones con parte nativa en Android, después comenta el código de la aplicación paso por paso incluyendo comentarios sobre los archivos adicionales necesarios para la compilación y finalmente detalla los archivos dedicados a la interfaz. Los experimentos incluyen una breve descripción sobre cómo interpretar los resultados seguidos de una serie de imágenes tomadas de la aplicación con diferentes localizaciones del patrón. En la entrega se incluye también un video. En el capítulo de resultados y conclusiones podemos encontrar observaciones sobre el desarrollo de la práctica, opiniones sobre su utilidad, y posibles mejoras.---ABSTRACT---This document is a guide that describes the development of and application for mobile devices in Android OS. The application combines computer vision techniques to calibrate the device camera and locate an element in the real world based on the parameters of the calibration The design of the application includes the decisions over the way that the application receives its input data, the patterns used in the calibration and localization and how the results are shown to the user. It also includes a flow chart that describes how the information travels along the application modules. The development begins with the steps necessary to configure the environment to develop native Android applications, then it explains the code step by step, including commentaries on the additional files necessary to build the application and details the files of the user interface. The experiments chapter explains the way the results are shown in the experiments before showing samples of different pattern localizations. There is also a video attached. In the conclusions chapter we can find observations on the development of the TFG, opinions about its usefulness, and possibilities of improvement in the future.

Caracterización multivariada de los temporales para su aplicación en el dimensionamiento del manto principal

En esta tesis se presenta una metodología para la caracterización del oleaje, dentro del marco de las nuevas Recomendaciones para Obras Marítimas (ROM 0.0.-00 y ROM 1.0-09), por ser una de las principales acciones que afectan a la estabilidad de las estructuras marítimas. Debido al carácter aleatorio intrínsecamente multivariado de la acción considerada, las tormentas, su caracterización paramétrica se realiza en términos de funciones cópula uniparamétricas. Las variables consideradas son altura de ola significante del pico de la tormenta, el periodo medio asociado y la magnitud, o número de olas, de todo el ciclo de solicitación. Para establecer un patrón teórico de evolución de la tormenta que permita extrapolar las muestras fuera de la región con datos se analizan los modelos teóricos existentes, comprobándose que no reproducen adecuadamente las tormentas constituidas por estados de mar con un peso importante de oleaje swell. Para evitar esta limitación se proponen cuatro modelos teóricos de evolución de tormentas con distintas formas geométricas. El análisis de los modelos existentes y los propuestos pone de relieve que el Modelo Magnitud Equivalente de Tormenta (EMS= Equivalent Magnitude Storm) con la forma triangular es el que mejor adapta las tormentas constituidas por estados de mar típicos del viento. Para tormentas con un mayor grado de desarrollo, el modelo teórico de tormenta EMS con la forma trapezoidal es el adecuado. De las aproximaciones propuestas para establecer el periodo medio de los sucesivos estados de mar del ciclo de solicitación. la propuesta por Martín Soldevilla et al., (2009) es la más versátil y , en general , mejor reproduce la evolución de todo tipo de tormentas. La caracterización de las tormentas se complementa con la altura de ola máxima. Debido a la mayor disponibilidad y longitud temporal de los datos sintéticos frente a las registros, la práctica totalidad de los análisis de extremos se realizan con tormentas sintéticas en las que la distribución de olas individuales es desconocida. Para evitar esta limitación se utilizan modelos teóricos de distribución de olas acordes a las características de cada uno de los estados de mar que conforman la tormenta sintética. Para establecer dichas características se utiliza la curtosis y en función de su valor la altura de ola máxima se determina asumiendo una determinada distribución de olas. Para estados de mar lineales la distribución de olas individuales de Rayleigh es la considerada. Para condiciones no lineales de gran ancho de banda el modelo de distribución de olas propuesto por Dawson, (2004) es el utilizado y si es de banda estrecha las predicciones de (Boccotti, (1989), Boccotti et al., (2013)) se compara con las resultantes del modelo de Dawson. La caracterización de la evolución de las tormentas en términos multivariados es aplicada al estudio de la progresión del daño del manto principal de diques en talud, y al rebase de las olas. Ambos aspectos cubren el segundo objetivo de la tesis en el que se propone una nueva formulación para el dimensionamiento de mantos constituidos por bloques cúbicos de hormigón. Para el desarrollo de esta nueva formulación se han utilizado los resultados recogidos en los estudios de estabilidad del manto principal de diques talud realizados en modelo físico a escala reducida en el Centro de Estudios de Puertos y Costas (CEDEX) desde la década de los 80 empleando, en su mayoría, bloques paralelepípedos cúbicos de hormigón. Por este motivo y porque los últimos diques construidos en la costa Española utilizan este tipo de pieza, es por lo que la formulación planteada se centra en este tipo de pieza. Después de un primer análisis de las fórmulas de cálculo y de evolución existentes, se llega a la conclusión de que es necesario realizar un esfuerzo de investigación en este campo, así como ensayos en laboratorio y recogida de datos in-situ con base a desarrollar fórmulas de evolución de daño para mantos constituidos por piezas diferentes a la escollera, que tenga en cuenta las principales variables que condiciona su estabilidad. En esta parte de la tesis se propone un método de análisis de evolución de daño, que incluye el criterio de inicio de avería, adecuada para diques en talud constituidos por bloque cúbicos de hormigón y que considera la incidencia oblicua, el daño acumulado y el rebase. This thesis proposes a methodology to estimate sea waves, one of the main actions affecting the maritime structures stability, complying with (ROM 0.0.-00 & ROM 1.0-09.Due to the multivariate behavior of sea storms, the characterization of the structures of sea storms is done using copula function. The analyzed variables are the significant height wave, mean period and magnitude or number of waves during the storm history. The storm evolution in terms of the significant height wave and the mean period is also studied in other to analyze the progressive failure modes. The existing models of evolution are studied, verifying that these approximations do not adjust accurately for developed waves. To overcome this disadvantage, four evolution models are proposed, with some geometrical shapes associated to fit any development degree. The proposed Equivalent Magnitude Storm model, EMS, generally obtains the best results for any kind of storm (predominant sea, swell or both). The triangle is recommended for typical sea storms whereas the trapezoid shape is much more appropriate for more developed storm conditions.The Martín Soldevilla et al., (2009) approach to estimate the mean period is better than others approaches used.The storm characterization is completed with the maximum wave height of the whole storm history. Due to synthetic historical waves databases are more accessible and longer than recorded database, the extreme analyses are done with synthetic data. For this reason the individual waves’ distribution is not known. For that limitation to be avoided, and depending on the characteristics of every sea states, one theoretical model of waves is choose and used. The kurtosis parameter is used to distinguish between linear and nonlinear sea states. The Rayleigh model is used for the linear sea states. For the nonlinear sea states, Dawson, (2004) approach is used for non-narrow bandwidth storms, comparing the results with the Boccotti, (1989), Boccotti et al., (2013) approach, with is used for narrow bandwidth storms. The multivariate and storm evolution characterization is used to analyze of stone armour damage progression and wave overtopping discharge. Both aspects are included in the second part of the thesis, with a new formula is proposed to design cubes armour layer. The results the stability studies of armour layer, done in the Centre for Harbours and Coastal Studies (CEDEX) laboratory are used for defining a new stability formula. For this reason and because the last biggest breakwater built in Spain using the cube, the damage progression is analyze for this kind of concrete block. Before to analyze the existing formulae, it is concluded that it is necessary more investigation, more tests in laboratory and data gathering in situ to define damage evolution formulae to armour of other kind of pieces and that takes to account the principal variables. This thesis proposed a method to calculate the damage progression including oblique waves, accumulated damage, and overtopping effect. The method also takes account the beginning of the movement of the blocks.

Nuevos desarrollos en la medida de descargas parciales y técnicas avanzadas de análisis para el diagnóstico del estado de los aislamientos en sistemas eléctricos de alta tensión

En esta tesis se desarrolla una investigación sobre las técnicas de medida de descargas parciales (DP) que se aplican en medidas on-line, en condiciones normales de operación de las instalaciones eléctricas de alta tensión (AT). También se realiza un estudio de técnicas avanzadas de procesado y análisis de las señales medidas, que permiten realizar diagnósticos precisos del estado de los aislamientos eléctricos de AT. Uno de los objetivos fundamentales de la tesis ha sido disponer de un procedimiento eficaz de medida y procesado de las señales de DP, para la realización de medidas on-line tanto de forma itinerante, como mediante monitorización temporal o permanente. La implementación del nuevo procedimiento de medida permite obtener resultados satisfactorios en la detección, identificación y localización de defectos de aislamiento. Se ha dedicado especial interés al desarrollo de un método de clasificación de señales, que permite separar pulsos de ruido y diferentes fuentes de DP, presentes de forma simultánea en las instalaciones de AT. El estudio de la clasificación de señales se ha completado con la aplicación de un método para la detección de manera asistida, de los diferentes grupos de pulsos de ruido y de DP. La aplicación de este método de detección de grupos de pulsos, facilita la labor de los técnicos especialistas a la hora de diagnosticar el estado de los elementos aislantes. Al efecto de verificar de forma práctica las aportaciones de la tesis, se han realizado medidas de DP tanto en laboratorio como en campo. Las medidas experimentales en laboratorio se han efectuado en el Laboratorio de Alta Tensión de la Escuela Técnica Superior de Ingeniería y Diseño Industrial (LAT-UPM), de la Universidad Politécnica de Madrid. Por otro lado, las medidas experimentales en campo se han llevado a cabo en instalaciones de AT propiedad de compañías de transporte y distribución de energía eléctrica. La realización de ensayos de DP en estas instalaciones ha sido posible, gracias a los proyectos de investigación llevados a cabo por el grupo de trabajo del LAT-UPM, con diferentes empresas del sector durante los diez últimos años. ABSTRACT This thesis develops techniques for measuring partial discharges (PD) that are applied in on-line measurements, under normal operating conditions of the high voltage (HV) electrical installations. In addition there are studied advanced techniques for the processing and analysis of the measured signals, that permit precise diagnostics of the state of HV electrical insulation systems. One of the fundamental objectives of the thesis is to make available an effective procedure for measuring and processing PD signals, for making on-line measurements, either in an itinerant way or in temporary or permanent monitoring. The implementation of the new measurement procedure yields satisfactory results in the detection, identification and localization of insulation defects. Special attention has been devoted to the development of a method for classifying signals, that separates noise pulses and various PD sources present simultaneously in the HV installations. The study of the classification of signals has been completed by the application of a method for detecting, in a user assisted manner, the different groups of noise pulses and of PD. The application of this method for detecting groups of pulses facilitates the work of the specialist technicians to diagnose the condition of the insulation elements. To demonstrate the practical value of the thesis, PD measurements were made in laboratory as well as in field installations. The experimental measurements in laboratory were made in the High Voltage Laboratory (LAT-UPM) of the High Technical School of Engineering and Industrial Design, of the Polytechnic University of Madrid. Field measurements were realized in the HV installations of companies providing electrical energy transport and distribution. The realization of PD tests in these facilities was possible thanks to the research projects carried out by the working group of the LAT-UPM during the last ten years, with different companies operating in the sector.

Monitorización de estructuras aeronáuticas mediante técnicas de inteligencia artificial

Una de las barreras para la aplicación de las técnicas de monitorización de la integridad estructural (SHM) basadas en ondas elásticas guiadas (GLW) en aeronaves es la influencia perniciosa de las condiciones ambientales y de operación (EOC). En esta tesis se ha estudiado dicha influencia y la compensación de la misma, particularizando en variaciones del estado de carga y temperatura. La compensación de dichos efectos se fundamenta en Redes Neuronales Artificiales (ANN) empleando datos experimentales procesados con la Transformada Chirplet. Los cambios en la geometría y en las propiedades del material respecto al estado inicial de la estructura (lo daños) provocan cambios en la forma de onda de las GLW (lo que denominamos característica sensible al daño o DSF). Mediante técnicas de tratamiento de señal se puede buscar una relación entre dichas variaciones y los daños, esto se conoce como SHM. Sin embargo, las variaciones en las EOC producen también cambios en los datos adquiridos relativos a las GLW (DSF) que provocan errores en los algoritmos de diagnóstico de daño (SHM). Esto sucede porque las firmas de daño y de las EOC en la DSF son del mismo orden. Por lo tanto, es necesario cuantificar y compensar el efecto de las EOC sobre la GLW. Si bien existen diversas metodologías para compensar los efectos de las EOC como por ejemplo “Optimal Baseline Selection” (OBS) o “Baseline Signal Stretching” (BSS), estas, se emplean exclusivamente en la compensación de los efectos térmicos. El método propuesto en esta tesis mezcla análisis de datos experimentales, como en el método OBS, y modelos basados en Redes Neuronales Artificiales (ANN) que reemplazan el modelado físico requerido por el método BSS. El análisis de datos experimentales consiste en aplicar la Transformada Chirplet (CT) para extraer la firma de las EOC sobre la DSF. Con esta información, obtenida bajo diversas EOC, se entrena una ANN. A continuación, la ANN actuará como un interpolador de referencias de la estructura sin daño, generando información de referencia para cualquier EOC. La comparación de las mediciones reales de la DSF con los valores simulados por la ANN, dará como resultado la firma daño en la DSF, lo que permite el diagnóstico de daño. Este esquema se ha aplicado y verificado, en diversas EOC, para una estructura unidimensional con un único camino de daño, y para una estructura representativa de un fuselaje de una aeronave, con curvatura y múltiples elementos rigidizadores, sometida a un estado de cargas complejo, con múltiples caminos de daños. Los efectos de las EOC se han estudiado en detalle en la estructura unidimensional y se han generalizado para el fuselaje, demostrando la independencia del método respecto a la configuración de la estructura y el tipo de sensores utilizados para la adquisición de datos GLW. Por otra parte, esta metodología se puede utilizar para la compensación simultánea de una variedad medible de EOC, que afecten a la adquisición de datos de la onda elástica guiada. El principal resultado entre otros, de esta tesis, es la metodología CT-ANN para la compensación de EOC en técnicas SHM basadas en ondas elásticas guiadas para el diagnóstico de daño. ABSTRACT One of the open problems to implement Structural Health Monitoring techniques based on elastic guided waves in real aircraft structures at operation is the influence of the environmental and operational conditions (EOC) on the damage diagnosis problem. This thesis deals with the compensation of these environmental and operational effects, specifically, the temperature and the external loading, by the use of the Chirplet Transform working with Artificial Neural Networks. It is well known that the guided elastic wave form is affected by the damage appearance (what is known as the damage sensitive feature or DSF). The DSF is modified by the temperature and by the load applied to the structure. The EOC promotes variations in the acquired data (DSF) and cause mistakes in damage diagnosis algorithms. This effect promotes changes on the waveform due to the EOC variations of the same order than the damage occurrence. It is difficult to separate both effects in order to avoid damage diagnosis mistakes. Therefore it is necessary to quantify and compensate the effect of EOC over the GLW forms. There are several approaches to compensate the EOC effects such as Optimal Baseline Selection (OBS) or Baseline Signal Stretching (BSS). Usually, they are used for temperature compensation. The new method proposed here mixes experimental data analysis, as in the OBS method, and Artificial Neural Network (ANN) models to replace the physical modelling which involves the BSS method. The experimental data analysis studied is based on apply the Chirplet Transform (CT) to extract the EOC signature on the DSF. The information obtained varying EOC is employed to train an ANN. Then, the ANN will act as a baselines interpolator of the undamaged structure. The ANN generates reference information at any EOC. By comparing real measurements of the DSF against the ANN simulated values, the damage signature appears clearly in the DSF, enabling an accurate damage diagnosis. This schema has been applied in a range of EOC for a one-dimensional structure containing single damage path and two dimensional real fuselage structure with stiffener elements and multiple damage paths. The EOC effects tested in the one-dimensional structure have been generalized to the fuselage showing its independence from structural arrangement and the type of sensors used for GLW data acquisition. Moreover, it can be used for the simultaneous compensation of a variety of measurable EOC, which affects the guided wave data acquisition. The main result, among others, of this thesis is the CT-ANN methodology for the compensation of EOC in GLW based SHM technique for damage diagnosis.

Activity-dependent regulation of synaptic clustering in a hippocampal culture system

Currently, there is a limited understanding of the factors that influence the localization and density of individual synapses in the central nervous system. Here we have studied the effects of activity on synapse formation between hippocampal dentate granule cells and CA3 pyramidal neurons in culture, taking advantage of FM1–43 as a fluorescent marker of synaptic boutons. We observed an early tendency for synapses to group together, quickly followed by the appearance of synaptic clusters on dendritic processes. These events were strongly influenced by N-methyl-d-aspartic acid receptor- and cyclic AMP-dependent signaling. The microstructure and localization of the synaptic clusters resembled that found in hippocampus, at mossy fiber synapses of stratum lucidum. Activity-dependent clustering of synapses represents a means for synaptic targeting that might contribute to synaptic organization in the brain.

Direct Visualization of the Human Estrogen Receptor α Reveals a Role for Ligand in the Nuclear Distribution of the Receptor

The human estrogen receptor α (ER α) has been tagged at its amino terminus with the S65T variant of the green fluorescent protein (GFP), allowing subcellular trafficking and localization to be observed in living cells by fluorescence microscopy. The tagged receptor, GFP-ER, is functional as a ligand-dependent transcription factor, responds to both agonist and antagonist ligands, and can associate with the nuclear matrix. Its cellular localization was analyzed in four human breast cancer epithelial cell lines, two ER+ (MCF7 and T47D) and two ER− (MDA-MB-231 and MDA-MB-435A), under a variety of ligand conditions. In all cell lines, GFP-ER is observed only in the nucleus in the absence of ligand. Upon the addition of agonist or antagonist ligand, a dramatic redistribution of GFP-ER from a reticular to punctate pattern occurs within the nucleus. In addition, the full antagonist ICI 182780 alters the nucleocytoplasmic compartmentalization of the receptor and causes partial accumulation in the cytoplasm in a process requiring continued protein synthesis. GFP-ER localization varies between cells, despite being cultured and treated in a similar manner. Analysis of the nuclear fluorescence intensity for variation in its frequency distribution helped establish localization patterns characteristic of cell line and ligand. During the course of this study, localization of GFP-ER to the nucleolar region is observed for ER− but not ER+ human breast cancer epithelial cell lines. Finally, our work provides a visual description of the “unoccupied” and ligand-bound receptor and is discussed in the context of the role of ligand in modulating receptor activity.

14–3-3 Inhibits the Dictyostelium Myosin II Heavy-Chain-specific Protein Kinase C Activity by a Direct Interaction: Identification of the 14–3-3 Binding Domain

Myosin II heavy chain (MHC) specific protein kinase C (MHC-PKC), isolated from Dictyostelium discoideum, regulates myosin II assembly and localization in response to the chemoattractant cyclic AMP. Immunoprecipitation of MHC-PKC revealed that it resides as a complex with several proteins. We show herein that one of these proteins is a homologue of the 14–3-3 protein (Dd14–3-3). This protein has recently been implicated in the regulation of intracellular signaling pathways via its interaction with several signaling proteins, such as PKC and Raf-1 kinase. We demonstrate that the mammalian 14–3-3 ζ isoform inhibits the MHC-PKC activity in vitro and that this inhibition is carried out by a direct interaction between the two proteins. Furthermore, we found that the cytosolic MHC-PKC, which is inactive, formed a complex with Dd14–3-3 in the cytosol in a cyclic AMP-dependent manner, whereas the membrane-bound active MHC-PKC was not found in a complex with Dd14–3-3. This suggests that Dd14–3-3 inhibits the MHC-PKC in vivo. We further show that MHC-PKC binds Dd14–3-3 as well as 14–3-3ζ through its C1 domain, and the interaction between these two proteins does not involve a peptide containing phosphoserine as was found for Raf-1 kinase. Our experiments thus show an in vivo function for a member of the 14–3-3 family and demonstrate that MHC-PKC interacts directly with Dd14–3-3 and 14–3-3ζ through its C1 domain both in vitro and in vivo, resulting in the inhibition of the kinase.

Luminal Heterodimeric Amino Acid Transporter Defective in Cystinuria

Mutations of the glycoprotein rBAT cause cystinuria type I, an autosomal recessive failure of dibasic amino acid transport (b0,+ type) across luminal membranes of intestine and kidney cells. Here we identify the permease-like protein b0,+AT as the catalytic subunit that associates by a disulfide bond with rBAT to form a hetero-oligomeric b0,+ amino acid transporter complex. We demonstrate its b0,+-type amino acid transport kinetics using a heterodimeric fusion construct and show its luminal brush border localization in kidney proximal tubule. These biochemical, transport, and localization characteristics as well as the chromosomal localization on 19q support the notion that the b0,+AT protein is the product of the gene defective in non-type I cystinuria.

Inhibition of β-catenin-mediated transactivation by cadherin derivatives

We studied the effect of N-cadherin, and its free or membrane-anchored cytoplasmic domain, on the level and localization of β-catenin and on its ability to induce lymphocyte enhancer-binding factor 1 (LEF-1)-responsive transactivation. These cadherin derivatives formed complexes with β-catenin and protected it from degradation. N-cadherin directed β-catenin into adherens junctions, and the chimeric protein induced diffuse distribution of β-catenin along the membrane whereas the cytoplasmic domain of N-cadherin colocalized with β-catenin in the nucleus. Cotransfection of β-catenin and LEF-1 into Chinese hamster ovary cells induced transactivation of a LEF-1 reporter, which was blocked by the N-cadherin-derived molecules. Expression of N-cadherin and an interleukin 2 receptor/cadherin chimera in SW480 cells relocated β-catenin from the nucleus to the plasma membrane and reduced transactivation. The cytoplasmic tails of N- or E-cadherin colocalized with β-catenin in the nucleus, and suppressed the constitutive LEF-1-mediated transactivation, by blocking β-catenin–LEF-1 interaction. Moreover, the 72 C-terminal amino acids of N-cadherin stabilized β-catenin and reduced its transactivation potential. These results indicate that β-catenin binding to the cadherin cytoplasmic tail either in the membrane, or in the nucleus, can inhibit β-catenin degradation and efficiently block its transactivation capacity.

Fluorescent RNA cytochemistry: tracking gene transcripts in living cells

The advent of jellyfish green fluorescent protein and its spectral variants, together with promising new fluorescent proteins from other classes of the Cnidarian phylum (coral and anemones), has greatly enhanced and promises to further boost the detection and localization of proteins in cell biology. It has been less widely appreciated that highly sensitive methods have also recently been developed for detecting the movement and localization in living cells of the very molecules that precede proteins in the gene expression pathway, i.e. RNAs. These approaches include the microinjection of fluorescent RNAs into living cells, the in vivo hybridization of fluorescent oligonucleotides to endogenous RNAs and the expression in cells of fluorescent RNA-binding proteins. This new field of ‘fluorescent RNA cytochemistry’ is summarized in this article, with emphasis on the biological insights it has already provided. These new techniques are likely to soon collaborate with other emerging approaches to advance the investigation of RNA birth, RNA–protein assembly and ribonucleoprotein particle transport in systems such as oocytes, embryos, neurons and other somatic cells, and may even permit the observation of viral replication and transcription pathways as they proceed in living cells, ushering in a new era of nucleic acids research in vivo.

Use of Ubiquitin Fusions to Augment Protein Expression in Transgenic Plants1

A major goal of plant biotechnology is the production of genetically engineered crops that express natural or foreign proteins at high levels. To enhance protein accumulation in transgenic plants, we developed a set of vectors that express proteins and peptides as C-terminal translational fusions with ubiquitin (UBQ). Studies of several proteins in tobacco (Nicotiana tabacum) showed that: (a) proteins can be readily expressed in plants as UBQ fusions; (b) by the action of endogenous UBQ-specific proteases (Ubps), these fusions are rapidly and precisely processed in vivo to release the fused protein moieties in free forms; (c) the synthesis of a protein as a UBQ fusion can significantly augment its accumulation; (d) proper processing and localization of a protein targeted to either the apoplast or the chloroplast is not affected by the N-terminal UBQ sequence; and (e) single amino acid substitutions surrounding the cleavage site can inhibit in vivo processing of the fusion by Ubps. Noncleavable UBQ fusions of β-glucuronidase became extensively modified, with additional UBQs in planta. Because multiubiquitinated proteins are the preferred substrates of the 26S proteasome, noncleavable fusions may be useful for decreasing protein half-life. Based on their ability to augment protein accumulation and the sequence specificity of Ubps, UBQ fusions offer a versatile way to express plant proteins.