Wind tunnel analysis of the flow topology over complex topographies

El principal objetivo de este trabajo es aportar conocimiento para contestar la pregunta: ¿hasta que punto los ensayos en túnel aerodinámico pueden contribuir a determinar las características que afectan la respuesta dinámica de los aerogeneradores operando en terreno complejo?. Esta pregunta no es nueva, de hecho, el debate en la comunidad científica comenzó en el primer tercio del siglo pasado y aún está intensamente vivo. El método generalmente aceptado para enfrentar el mencionado problema consiste en analizar un caso de estudio determinado en el cual se aplican tanto ensayos a escala real como análisis computacionales y ensayos en túnel aerodinámico. Esto no es ni fácil ni barato. Esta es la razón por la cual desde el experimento de Askervein en 1988, los modelizadores del flujo atmosférico tuvieron que esperar hasta 2007 a que el experimento de Bolund fuese puesto en marcha con un despliegue de medios técnicos equivalentes (teniendo en cuenta la evolución de las tecnologías de sensores y computación). El problema contempla tantos aspectos que ambas experiencias fueron restringidas a condiciones de atmósfera neutra con efectos de Coriolis despreciables con objeto de reducir la complejidad. Este es el contexto en el que se ha desarrollado la presente tesis doctoral. La topología del flujo sobre la isla de Bolund ha sido estudiada mediante la reproducción del experimento de Bolund en los túneles aerodinámicos A9 y ACLA16 del IDR. Dos modelos de la isla de Bolund fueron fabricados a dos escalas, 1:230 y 1:115. El flujo de entrada en el túnel aerodinámico simulando la capa límite sin perturbar correspondía a régimen de transición (transitionally rough regime) y fue usado como situación de referencia. El modelo a escala 1:230 fue ensayado en el túnel A9 para determinar la presión sobre su superficie. La distribución del coeficiente de presión sobre la isla proporcionó una visualización y estimación de una región de desprendimiento sobre el pequeño acantilado situado al frente de la misma. Las medidas de presión instantánea con suficiente grado de resolución temporal pusieron de manifiesto la no estacionariedad en la región de desprendimiento. El modelo a escala 1:115 fue ensayado utilizando hilo caliente de tres componentes y un sistema de velocimetría por imágenes de partículas de dos componentes. El flujo fue caracterizado por el ratio de aceleración, el incremento normalizado de energía cinética turbulenta y los ángulos de inclinación y desviación horizontal. Los resultados a lo largo de la dirección 270°y alturas de 2 m y 5 m presentaron una gran similitud con los resultados a escala real del experimento de Bolund. Los perfiles verticales en las localizaciones de las torres meteorológicas mostraron un acuerdo significativo con los resultados a escala real. El análisis de los esfuerzos de Reynolds y el análisis espectral en las localizaciones de los mástiles meteorológicos presentaron niveles de acuerdo variados en ciertas posiciones, mientras que en otras presentaron claras diferencias. El mapeo horizontal del flujo, para una dirección de viento de 270°, permitió caracterizar el comportamiento de la burbuja intermitente de recirculación sobre el pequeño acantilado existente al frente de la isla así como de la región de relajación y de la capa de cortadura en la región corriente abajo de Bolund. Se realizaron medidas de velocidad con alta resolución espacial en planos perpendiculares a la dirección del flujo sin perturbar. Estas medidas permitieron detectar y caracterizar una estructura de flujo similar a un torbellino longitudinal con regiones con altos gradientes de velocidad y alta intensidad de turbulencia. Esta estructura de flujo es, sin duda, un reto para los modelos computacionales y puede considerarse un factor de riesgo para la operación de los aerogeneradores. Se obtuvieron y analizaron distribuciones espaciales de los esfuerzos de Reynolds mediante 3CHW y PIV. Este tipo de parámetros no constituyen parte de los resultados habituales en los ensayos en túnel sobre topografías y son muy útiles para los modelizadores que utilizan simulación de grades torbellinos (LES). Se proporciona una interpretación de los resultados obtenidos en el túnel aerodinámico en términos de utilidad para los diseñadores de parques eólicos. La evolución y variación de los parámetros del flujo a lo largo de líneas, planos y superficies han permitido identificar como estas propiedades del flujo podrían afectar la localización de los aerogeneradores y a la clasificación de emplazamientos. Los resultados presentados sugieren, bajo ciertas condiciones, la robustez de los ensayos en túnel para estudiar la topología sobre terreno complejo y su comparabilidad con otras técnicas de simulación, especialmente considerando el nivel de acuerdo del conjunto de resultados presentados con los resultados a escala real. De forma adicional, algunos de los parámetros del flujo obtenidos de las medidas en túnel son difícilmente determinables en ensayos a escala real o por medios computacionales, considerado el estado del arte. Este trabajo fue realizado como parte de las actividades subvencionadas por la Comisión Europea como dentro del proyecto FP7-PEOPLE-ITN-2008WAUDIT (Wind Resource Assessment Audit and Standardization) dentro de la FP7 Marie-Curie Initial Training Network y por el Ministerio Español de Economía y Competitividad dentro del proyecto ENE2012-36473, TURCO (Determinación en túnel aerodinámico de la distribución espacial de parámetros estadísticos de la turbulencia atmosférica sobre topografías complejas) del Plan Nacional de Investigación (Subprograma de investigación fundamental no orientada 2012). El informe se ha organizado en siete capítulos y un conjunto de anexos. En el primer capítulo se introduce el problema. En el capítulo dos se describen los medios experimentales utilizados. Seguidamente, en el capítulo tres, se analizan en detalle las condiciones de referencia del principal túnel aerodinámico utilizado en esta investigación. En el capítulo tres se presentan resultados de ensayos de presión superficial sobre un modelo de la isla. Los principales resultados del experimento de Bolund se reproducen en el capítulo cinco. En el capítulo seis se identifican diferentes estructuras del flujo sobre la isla y, finalmente, en el capitulo siete, se recogen las conclusiones y una propuesta de lineas de trabajo futuras. ABSTRACT The main objective of this work is to contribute to answer the question: to which extend can the wind tunnel testing contribute to determine the flow characteristics that affect the dynamic response of wind turbines operating in highly complex terrains?. This question is not new, indeed, the debate in the scientific community was opened in the first third of the past century and it is still intensely alive. The accepted approach to face this problem consists in analysing a given case study where full-scale tests, computational modelling and wind tunnel testing are applied to the same topography. This is neither easy nor cheap. This is is the reason why since the Askervein experience in 1988, the atmospheric flow modellers community had to wait till 2007 when the Bolund experiment was setup with a deployment of technical means equivalent (considering the evolution of the sensor and computing techniques). The problem is so manifold that both experiences were restricted to neutral conditions without Coriolis effects in order to reduce the complexity. This is the framework in which this PhD has been carried out. The flow topology over the Bolund Island has been studied by replicating the Bolund experiment in the IDR A9 and ACLA16 wind tunnels. Two mock-ups of the Bolund island were manufactured at two scales of 1:230 and 1:115. The in-flow in the empty wind tunnel simulating the incoming atmospheric boundary layer was in the transitionally rough regime and used as a reference case. The 1:230 model was tested in the A9 wind tunnel to measure surface pressure. The mapping of the pressure coefficient across the island gave a visualisation and estimation of a detachment region on the top of the escarpment in front of the island. Time resolved instantaneous pressure measurements illustrated the non-steadiness in the detachment region. The 1:115 model was tested using 3C hot-wires(HW) and 2C Particle Image Velocimetry(PIV). Measurements at met masts M3, M6, M7 and M8 and along Line 270°were taken to replicate the result of the Bolund experiment. The flow was characterised by the speed-up ratio, normalised increment of the turbulent kinetic energy, inclination angle and turning angle. Results along line 270°at heights of 2 m and 5 m compared very well with the full-scale results of the Bolund experiment. Vertical profiles at the met masts showed a significant agreement with the full-scale results. The analysis of the Reynolds stresses and the spectral analysis at the met mast locations gave a varied level of agreement at some locations while clear mismatch at others. The horizontal mapping of the flow field, for a 270°wind direction, allowed to characterise the behaviour of the intermittent recirculation bubble on top of the front escarpment followed by a relaxation region and the presence of a shear layer in the lee side of the island. Further detailed velocity measurements were taken at cross-flow planes over the island to study the flow structures on the island. A longitudinal vortex-like structure with high mean velocity gradients and high turbulent kinetic energy was characterised on the escarpment and evolving downstream. This flow structure is a challenge to the numerical models while posing a threat to wind farm designers when siting wind turbines. Spatial distribution of Reynold stresses were presented from 3C HW and PIV measurements. These values are not common results from usual wind tunnel measurements and very useful for modellers using large eddy simulation (LES). An interpretation of the wind tunnel results in terms of usefulness to wind farm designers is given. Evolution and variation of the flow parameters along measurement lines, planes and surfaces indicated how the flow field could affect wind turbine siting. Different flow properties were presented so compare the level of agreement to full-scale results and how this affected when characterising the site wind classes. The results presented suggest, under certain conditions, the robustness of the wind tunnel testing for studying flow topology over complex terrain and its capability to compare to other modelling techniques especially from the level of agreement between the different data sets presented. Additionally, some flow parameters obtained from wind tunnel measurements would have been quite difficult to be measured at full-scale or by computational means considering the state of the art. This work was carried out as a part of the activities supported by the EC as part of the FP7- PEOPLE-ITN-2008 WAUDIT project (Wind Resource Assessment Audit and Standardization) within the FP7 Marie-Curie Initial Training Network and by the Spanish Ministerio de Economía y Competitividad, within the framework of the ENE2012-36473, TURCO project (Determination of the Spatial Distribution of Statistic Parameters of Flow Turbulence over Complex Topographies in Wind Tunnel) belonging to the Spanish National Program of Research (Subprograma de investigación fundamental no orientada 2012). The report is organised in seven chapters and a collection of annexes. In chapter one, the problem is introduced. In chapter two the experimental setup is described. Following, in chapter three, the inflow conditions of the main wind tunnel used in this piece of research are analysed in detail. In chapter three, preliminary pressure tests results on a model of the island are presented. The main results from the Bolund experiment are replicated in chapter five. In chapter six, an identification of specific flow strutures over the island is presented and, finally, in chapter seven, conclusions and lines for future works related to the presented one are included.

Improving parameters selection of a seeded region growing method for multiband image segmentation

In the last decade, Object Based Image Analysis (OBIA) has been accepted as an effective method for processing high spatial resolution multiband images. This image analysis method is an approach that starts with the segmentation of the image. Image segmentation in general is a procedure to partition an image into homogenous groups (segments). In practice, visual interpretation is often used to assess the quality of segmentation and the analysis relies on the experience of an analyst. In an effort to address the issue, in this study, we evaluate several seed selection strategies for an automatic image segmentation methodology based on a seeded region growing-merging approach. In order to evaluate the segmentation quality, segments were subjected to spatial autocorrelation analysis using Moran's I index and intra-segment variance analysis. We apply the algorithm to image segmentation using an aerial multiband image.

Detection and quantification of peanut traces in wheat flour by near infrared hyperspectral imaging spectroscopy using principal-component analysis

The use of a common environment for processing different powder foods in the industry has increased the risk of finding peanut traces in powder foods. The analytical methods commonly used for detection of peanut such as enzyme-linked immunosorbent assay (ELISA) and real-time polymerase chain reaction (RT-PCR) represent high specificity and sensitivity but are destructive and time-consuming, and require highly skilled experimenters. The feasibility of NIR hyperspectral imaging (HSI) is studied for the detection of peanut traces down to 0.01% by weight. A principal-component analysis (PCA) was carried out on a dataset of peanut and flour spectra. The obtained loadings were applied to the HSI images of adulterated wheat flour samples with peanut traces. As a result, HSI images were reduced to score images with enhanced contrast between peanut and flour particles. Finally, a threshold was fixed in score images to obtain a binary classification image, and the percentage of peanut adulteration was compared with the percentage of pixels identified as peanut particles. This study allowed the detection of traces of peanut down to 0.01% and quantification of peanut adulteration from 10% to 0.1% with a coefficient of determination (r2) of 0.946. These results show the feasibility of using HSI systems for the detection of peanut traces in conjunction with chemical procedures, such as RT-PCR and ELISA to facilitate enhanced quality-control surveillance on food-product processing lines.

Quantitative analysis of morphological changes in yeast colonies growing on solid medium: the eccentricity and Fourier indices

The colony shape of four yeast species growing on agar medium wasmeasured for 116 days by image analysis. Initially, all the colonies are circular, with regular edges. The loss of circularity can be quantitatively estimated by the eccentricity index, Ei, calculated as the ratio between their orthogonal vertical and horizontal diameters. Ei can increase from 1 (complete circularity) to a maximum of 1.17–1.30, depending on the species. One colony inhibits its neighbour only when it has reached a threshold area. Then, Ei of the inhibited colony increases proportionally to the area of the inhibitory colony. The initial distance between colonies affects those threshold values but not the proportionality, Ei/area; this inhibition affects the shape but not the total surface of the colony. The appearance of irregularities in the edges is associated, in all the species, not with age but with nutrient exhaustion. The edge irregularity can be quantified by the Fourier index, Fi, calculated by the minimum number of Fourier coefficients that are needed to describe the colony contour with 99% fitness. An ad hoc function has been developed in Matlab v. 7.0 to automate the computation of the Fourier coefficients. In young colonies, Fi has a value between 2 (circumference) and 3 (ellipse). These values are maintained in mature colonies of Debaryomyces, but can reach values up to 14 in Saccharomyces.All the species studied showed the inhibition of growth in facing colony edges, but only three species showed edge irregularities associated with substrate exhaustion. Copyright © 2014 John Wiley & Sons, Ltd.

Systematic analysis of the decoding delay in multiview video

We present a framework for the analysis of the decoding delay in multiview video coding (MVC). We show that in real-time applications, an accurate estimation of the decoding delay is essential to achieve a minimum communication latency. As opposed to single-view codecs, the complexity of the multiview prediction structure and the parallel decoding of several views requires a systematic analysis of this decoding delay, which we solve using graph theory and a model of the decoder hardware architecture. Our framework assumes a decoder implementation in general purpose multi-core processors with multi-threading capabilities. For this hardware model, we show that frame processing times depend on the computational load of the decoder and we provide an iterative algorithm to compute jointly frame processing times and decoding delay. Finally, we show that decoding delay analysis can be applied to design decoders with the objective of minimizing the communication latency of the MVC system.

A study of feature combination for vehicle detection based on image processing

Video analytics play a critical role in most recent traffic monitoring and driver assistance systems. In this context, the correct detection and classification of surrounding vehicles through image analysis has been the focus of extensive research in the last years. Most of the pieces of work reported for image-based vehicle verification make use of supervised classification approaches and resort to techniques, such as histograms of oriented gradients (HOG), principal component analysis (PCA), and Gabor filters, among others. Unfortunately, existing approaches are lacking in two respects: first, comparison between methods using a common body of work has not been addressed; second, no study of the combination potentiality of popular features for vehicle classification has been reported. In this study the performance of the different techniques is first reviewed and compared using a common public database. Then, the combination capabilities of these techniques are explored and a methodology is presented for the fusion of classifiers built upon them, taking into account also the vehicle pose. The study unveils the limitations of single-feature based classification and makes clear that fusion of classifiers is highly beneficial for vehicle verification.

Methods for the analysis of multi-scale cell dynamics from fluorescence microscopy images

La embriogénesis es el proceso mediante el cual una célula se convierte en un ser un vivo. A lo largo de diferentes etapas de desarrollo, la población de células va proliferando a la vez que el embrión va tomando forma y se configura. Esto es posible gracias a la acción de varios procesos genéticos, bioquímicos y mecánicos que interaccionan y se regulan entre ellos formando un sistema complejo que se organiza a diferentes escalas espaciales y temporales. Este proceso ocurre de manera robusta y reproducible, pero también con cierta variabilidad que permite la diversidad de individuos de una misma especie. La aparición de la microscopía de fluorescencia, posible gracias a proteínas fluorescentes que pueden ser adheridas a las cadenas de expresión de las células, y los avances en la física óptica de los microscopios han permitido observar este proceso de embriogénesis in-vivo y generar secuencias de imágenes tridimensionales de alta resolución espacio-temporal. Estas imágenes permiten el estudio de los procesos de desarrollo embrionario con técnicas de análisis de imagen y de datos, reconstruyendo dichos procesos para crear la representación de un embrión digital. Una de las más actuales problemáticas en este campo es entender los procesos mecánicos, de manera aislada y en interacción con otros factores como la expresión genética, para que el embrión se desarrolle. Debido a la complejidad de estos procesos, estos problemas se afrontan mediante diferentes técnicas y escalas específicas donde, a través de experimentos, pueden hacerse y confrontarse hipótesis, obteniendo conclusiones sobre el funcionamiento de los mecanismos estudiados. Esta tesis doctoral se ha enfocado sobre esta problemática intentando mejorar las metodologías del estado del arte y con un objetivo específico: estudiar patrones de deformación que emergen del movimiento organizado de las células durante diferentes estados del desarrollo del embrión, de manera global o en tejidos concretos. Estudios se han centrado en la mecánica en relación con procesos de señalización o interacciones a nivel celular o de tejido. En este trabajo, se propone un esquema para generalizar el estudio del movimiento y las interacciones mecánicas que se desprenden del mismo a diferentes escalas espaciales y temporales. Esto permitiría no sólo estudios locales, si no estudios sistemáticos de las escalas de interacción mecánica dentro de un embrión. Por tanto, el esquema propuesto obvia las causas de generación de movimiento (fuerzas) y se centra en la cuantificación de la cinemática (deformación y esfuerzos) a partir de imágenes de forma no invasiva. Hoy en día las dificultades experimentales y metodológicas y la complejidad de los sistemas biológicos impiden una descripción mecánica completa de manera sistemática. Sin embargo, patrones de deformación muestran el resultado de diferentes factores mecánicos en interacción con otros elementos dando lugar a una organización mecánica, necesaria para el desarrollo, que puede ser cuantificado a partir de la metodología propuesta en esta tesis. La metodología asume un medio continuo descrito de forma Lagrangiana (en función de las trayectorias de puntos materiales que se mueven en el sistema en lugar de puntos espaciales) de la dinámica del movimiento, estimado a partir de las imágenes mediante métodos de seguimiento de células o de técnicas de registro de imagen. Gracias a este esquema es posible describir la deformación instantánea y acumulada respecto a un estado inicial para cualquier dominio del embrión. La aplicación de esta metodología a imágenes 3D + t del pez zebra sirvió para desvelar estructuras mecánicas que tienden a estabilizarse a lo largo del tiempo en dicho embrión, y que se organizan a una escala semejante al del mapa de diferenciación celular y con indicios de correlación con patrones de expresión genética. También se aplicó la metodología al estudio del tejido amnioserosa de la Drosophila (mosca de la fruta) durante el cierre dorsal, obteniendo indicios de un acoplamiento entre escalas subcelulares, celulares y supracelulares, que genera patrones complejos en respuesta a la fuerza generada por los esqueletos de acto-myosina. En definitiva, esta tesis doctoral propone una estrategia novedosa de análisis de la dinámica celular multi-escala que permite cuantificar patrones de manera inmediata y que además ofrece una representación que reconstruye la evolución de los procesos como los ven las células, en lugar de como son observados desde el microscopio. Esta metodología por tanto permite nuevas formas de análisis y comparación de embriones y tejidos durante la embriogénesis a partir de imágenes in-vivo. ABSTRACT The embryogenesis is the process from which a single cell turns into a living organism. Through several stages of development, the cell population proliferates at the same time the embryo shapes and the organs develop gaining their functionality. This is possible through genetic, biochemical and mechanical factors that are involved in a complex interaction of processes organized in different levels and in different spatio-temporal scales. The embryogenesis, through this complexity, develops in a robust and reproducible way, but allowing variability that makes possible the diversity of living specimens. The advances in physics of microscopes and the appearance of fluorescent proteins that can be attached to expression chains, reporting about structural and functional elements of the cell, have enabled for the in-vivo observation of embryogenesis. The imaging process results in sequences of high spatio-temporal resolution 3D+time data of the embryogenesis as a digital representation of the embryos that can be further analyzed, provided new image processing and data analysis techniques are developed. One of the most relevant and challenging lines of research in the field is the quantification of the mechanical factors and processes involved in the shaping process of the embryo and their interactions with other embryogenesis factors such as genetics. Due to the complexity of the processes, studies have focused on specific problems and scales controlled in the experiments, posing and testing hypothesis to gain new biological insight. However, methodologies are often difficult to be exported to study other biological phenomena or specimens. This PhD Thesis is framed within this paradigm of research and tries to propose a systematic methodology to quantify the emergent deformation patterns from the motion estimated in in-vivo images of embryogenesis. Thanks to this strategy it would be possible to quantify not only local mechanisms, but to discover and characterize the scales of mechanical organization within the embryo. The framework focuses on the quantification of the motion kinematics (deformation and strains), neglecting the causes of the motion (forces), from images in a non-invasive way. Experimental and methodological challenges hamper the quantification of exerted forces and the mechanical properties of tissues. However, a descriptive framework of deformation patterns provides valuable insight about the organization and scales of the mechanical interactions, along the embryo development. Such a characterization would help to improve mechanical models and progressively understand the complexity of embryogenesis. This framework relies on a Lagrangian representation of the cell dynamics system based on the trajectories of points moving along the deformation. This approach of analysis enables the reconstruction of the mechanical patterning as experienced by the cells and tissues. Thus, we can build temporal profiles of deformation along stages of development, comprising both the instantaneous events and the cumulative deformation history. The application of this framework to 3D + time data of zebrafish embryogenesis allowed us to discover mechanical profiles that stabilized through time forming structures that organize in a scale comparable to the map of cell differentiation (fate map), and also suggesting correlation with genetic patterns. The framework was also applied to the analysis of the amnioserosa tissue in the drosophila’s dorsal closure, revealing that the oscillatory contraction triggered by the acto-myosin network organized complexly coupling different scales: local force generation foci, cellular morphology control mechanisms and tissue geometrical constraints. In summary, this PhD Thesis proposes a theoretical framework for the analysis of multi-scale cell dynamics that enables to quantify automatically mechanical patterns and also offers a new representation of the embryo dynamics as experienced by cells instead of how the microscope captures instantaneously the processes. Therefore, this framework enables for new strategies of quantitative analysis and comparison between embryos and tissues during embryogenesis from in-vivo images.

Analysis by finite element calculations of light scattering in laser-textured AZO films for PV thin-film solar cells

In the thin-film photovoltaic industry, to achieve a high light scattering in one or more of the cell interfaces is one of the strategies that allow an enhancement of light absorption inside the cell and, therefore, a better device behavior and efficiency. Although chemical etching is the standard method to texture surfaces for that scattering improvement, laser light has shown as a new way for texturizing different materials, maintaining a good control of the final topography with a unique, clean, and quite precise process. In this work AZO films with different texture parameters are fabricated. The typical parameters used to characterize them, as the root mean square roughness or the haze factor, are discussed and, for deeper understanding of the scattering mechanisms, the light behavior in the films is simulated using a finite element method code. This method gives information about the light intensity in each point of the system, allowing the precise characterization of the scattering behavior near the film surface, and it can be used as well to calculate a simulated haze factor that can be compared with experimental measurements. A discussion of the validation of the numerical code, based in a comprehensive comparison with experimental data is included.

Analysis of the role of the Spitzenkörper in fungal morphogenesis by computer simulation of apical branching in Aspergillus niger

High-resolution video microscopy, image analysis, and computer simulation were used to study the role of the Spitzenkörper (Spk) in apical branching of ramosa-1, a temperature-sensitive mutant of Aspergillus niger. A shift to the restrictive temperature led to a cytoplasmic contraction that destabilized the Spk, causing its disappearance. After a short transition period, new Spk appeared where the two incipient apical branches emerged. Changes in cell shape, growth rate, and Spk position were recorded and transferred to the fungus simulator program to test the hypothesis that the Spk functions as a vesicle supply center (VSC). The simulation faithfully duplicated the elongation of the main hypha and the two apical branches. Elongating hyphae exhibited the growth pattern described by the hyphoid equation. During the transition phase, when no Spk was visible, the growth pattern was nonhyphoid, with consecutive periods of isometric and asymmetric expansion; the apex became enlarged and blunt before the apical branches emerged. Video microscopy images suggested that the branch Spk were formed anew by gradual condensation of vesicle clouds. Simulation exercises where the VSC was split into two new VSCs failed to produce realistic shapes, thus supporting the notion that the branch Spk did not originate by division of the original Spk. The best computer simulation of apical branching morphogenesis included simulations of the ontogeny of branch Spk via condensation of vesicle clouds. This study supports the hypothesis that the Spk plays a major role in hyphal morphogenesis by operating as a VSC—i.e., by regulating the traffic of wall-building vesicles in the manner predicted by the hyphoid model.

Crystal structure of D-amino acid oxidase: a case of active site mirror-image convergent evolution with flavocytochrome b2.

D-amino acid oxidase is the prototype of the FAD-dependent oxidases. It catalyses the oxidation of D-amino acids to the corresponding alpha-ketoacids. The reducing equivalents are transferred to molecular oxygen with production of hydrogen peroxide. We have solved the crystal structure of the complex of D-amino acid oxidase with benzoate, a competitive inhibitor of the substrate, by single isomorphous replacement and eightfold averaging. Each monomer is formed by two domains with an overall topology similar to that of p-hydroxybenzoate hydroxylase. The benzoate molecule lays parallel to the flavin ring and is held in position by a salt bridge with Arg-283. Analysis of the active site shows that no side chains are properly positioned to act as the postulated base required for the catalytic carboanion mechanism. On the contrary, the benzoate binding mode suggests a direct transfer of the substrate alpha-hydrogen to the flavin during the enzyme reductive half-reaction.The active site Of D-amino acid oxidase exhibits a striking similarity with that of flavocytochrome b2, a structurally unrelated FMN-dependent flavoenzyme. The active site groups (if these two enzymes are in fact superimposable once the mirror-image of the flavocytochrome b2 active site is generated with respect to the flavin plane. Therefore, the catalytic sites of D-amino acid oxidase and flavocytochrome b2 appear to have converged to a highly similar but enantiomeric architecture in order to catalvze similar reactions (oxidation of alpha-amino acids or alpha-hydroxy acids), although with opposite stereochemistry.

Quantitative analysis of senile plaques in Alzheimer disease: observation of log-normal size distribution and molecular epidemiology of differences associated with apolipoprotein E genotype and trisomy 21 (Down syndrome).

The discovery that the epsilon 4 allele of the apolipoprotein E (apoE) gene is a putative risk factor for Alzheimer disease (AD) in the general population has highlighted the role of genetic influences in this extremely common and disabling illness. It has long been recognized that another genetic abnormality, trisomy 21 (Down syndrome), is associated with early and severe development of AD neuropathological lesions. It remains a challenge, however, to understand how these facts relate to the pathological changes in the brains of AD patients. We used computerized image analysis to examine the size distribution of one of the characteristic neuropathological lesions in AD, deposits of A beta peptide in senile plaques (SPs). Surprisingly, we find that a log-normal distribution fits the SP size distribution quite well, motivating a porous model of SP morphogenesis. We then analyzed SP size distribution curves in genotypically defined subgroups of AD patients. The data demonstrate that both apoE epsilon 4/AD and trisomy 21/AD lead to increased amyloid deposition, but by apparently different mechanisms. The size distribution curve is shifted toward larger plaques in trisomy 21/AD, probably reflecting increased A beta production. In apoE epsilon 4/AD, the size distribution is unchanged but the number of SP is increased compared to apoE epsilon 3, suggesting increased probability of SP initiation. These results demonstrate that subgroups of AD patients defined on the basis of molecular characteristics have quantitatively different neuropathological phenotypes.