Evolución del conocimiento del hormigón estructural hasta 1973

Esta tesis analiza los criterios con que fueron proyectadas y construidas las estructuras de hormigón hasta 1973, fecha coincidente con la Instrucción EH-73, que en contenido, formato y planteamiento, consagró la utilización de los criterios modernamente utilizados hasta ahora. Es heredera, además, de las CEB 1970. Esos años marcan el cambio de planteamiento desde la Teoría Clásica hacia los Estados Límite. Los objetivos perseguidos son, sintéticamente: 1) Cubrir un vacío patente en el estudio de la evolución del conocimiento. Hay tratados sobre la historia del hormigón que cubren de manera muy completa el relato de personajes y realizaciones, pero no, al menos de manera suficiente, la evolución del conocimiento. 2) Servir de ayuda a los técnicos de hoy para entender configuraciones estructurales, geometrías, disposiciones de armado, formatos de seguridad, etc, utilizados en el pasado, lo que servirá para la redacción más fundada de dictámenes preliminares sobre estructuras existentes. 3) Ser referencia para la realización de estudios de valoración de la capacidad resistente de construcciones existentes, constituyendo la base de un documento pre-normativo orientado en esa dirección. En efecto, esta tesis pretende ser una ayuda para los ingenieros de hoy que se enfrentan a la necesidad de conservar y reparar estructuras de hormigón armado que forman parte del patrimonio heredado. La gran mayoría de las estructuras, fueron construidas hace más de 40 años, por lo que es preciso conocer los criterios que marcaron su diseño, su cálculo y su construcción. Pretende determinar cuáles eran los límites de agotamiento y por tanto de seguridad, de estructuras dimensionadas con criterios de antaño, analizadas por la metodología de cálculo actual. De este modo, se podrá determinar el resguardo existente “real” de las estructuras dimensionadas y calculadas con criterios “distintos” a los actuales. Conocer el comportamiento de las estructuras construidas con criterios de la Teoría Clásica, según los criterios actuales, permitirá al ingeniero de hoy tratar de la forma más adecuada el abanico de necesidades que se puedan presentar en una estructura existente. Este trabajo se centra en la evolución del conocimiento por lo que no se encuentran incluidos los procesos constructivos. En lo relativo a los criterios de proyecto, hasta mediados del siglo XX, éstos se veían muy influidos por los ensayos y trabajos de autor consiguientes, en los que se basaban los reglamentos de algunos países. Era el caso del reglamento prusiano de 1904, de la Orden Circular francesa de 1906, del Congreso de Lieja de 1930. A partir de la segunda mitad del siglo XX, destacan las aportaciones de ingenieros españoles como es el caso de Alfredo Páez Balaca, Eduardo Torroja y Pedro Jiménez Montoya, entre otros, que permitieron el avance de los criterios de cálculo y de seguridad de las estructuras de hormigón, hasta los que se conocen hoy. El criterio rector del proyecto de las estructuras de hormigón se fundó, como es sabido, en los postulados de la Teoría Clásica, en particular en el “momento crítico”, aquel para el que hormigón y acero alcanzan sus tensiones admisibles y, por tanto, asegura el máximo aprovechamiento de los materiales y sin pretenderlo conscientemente, la máxima ductilidad. Si el momento solicitante es mayor que el crítico, se dispone de armadura en compresión. Tras el estudio de muchas de las estructuras existentes de la época por el autor de esta tesis, incluyendo entre ellas las Colecciones Oficiales de Puentes de Juan Manuel de Zafra, Eugenio Ribera y Carlos Fernández Casado, se concluye que la definición geométrica de las mismas no se corresponde exactamente con la resultante del momento crítico, dado que como ahora resultaba necesario armonizar los criterios de armado a nivel sección con la organización de la ferralla a lo largo de los diferentes elementos estructurales. Los parámetros de cálculo, resistencias de los materiales y formatos de seguridad, fueron evolucionando con los años. Se fueron conociendo mejor las prestaciones de los materiales, se fue enriqueciendo la experiencia de los propios procesos constructivos y, en menor medida, de las acciones solicitantes y, consiguientemente, acotándose las incertidumbres asociadas lo cual permitió ir ajustando los coeficientes de seguridad a emplear en el cálculo. Por ejemplo, para el hormigón se empleaba un coeficiente de seguridad igual a 4 a finales del siglo XIX, que evolucionó a 3,57 tras la publicación de la Orden Circular francesa de 1906, y a 3, tras la Instrucción española de 1939. En el caso del acero, al ser un material bastante más conocido por cuanto se había utilizado muchísimo previamente, el coeficiente de seguridad permaneció casi constante a lo largo de los años, con un valor igual a 2. Otra de las causas de la evolución de los parámetros de cálculo fue el mejor conocimiento del comportamiento de las estructuras merced a la vasta tarea de planificación y ejecución de ensayos, con los estudios teóricos consiguientes, realizados por numerosos autores, principalmente austríacos y alemanes, pero también norteamericanos y franceses. En cuanto a los criterios de cálculo, puede sorprender al técnico de hoy el conocimiento que tenían del comportamiento del hormigón desde los primeros años del empleo del mismo. Sabían del comportamiento no lineal del hormigón, pero limitaban su trabajo a un rango de tensióndeformación lineal porque eso aseguraba una previsión del comportamiento estructural conforme a las hipótesis de la Elasticidad Lineal y de la Resistencia de Materiales, muy bien conocidas a principios del s. XX (no así sucedía con la teoría de la Plasticidad, aún sin formular, aunque estaba implícita en los planteamientos algunos ingenieros especializados en estructuras de fábrica (piedra o ladrillo) y metálicas. Además, eso permitía independizar un tanto el proyecto de los valores de las resistencias reales de los materiales, lo que liberaba de la necesidad de llevar a cabo ensayos que, en la práctica, apenas se podían hacer debido a la escasez de los laboratorios. Tampoco disponían de programas informáticos ni de ninguna de las facilidades de las que hoy se tienen, que les permitiera hacer trabajar al hormigón en un rango no lineal. Así, sabia y prudentemente, limitaban las tensiones y deformaciones del material a un rango conocido. El modus operandi seguido para la elaboración de esta tesis, ha sido el siguiente: -Estudio documental: se han estudiado documentos de autor, recomendaciones y normativa generada en este ámbito, tanto en España como con carácter internacional, de manera sistemática con arreglo al índice del documento. En este proceso, se han detectado lagunas del conocimiento (y su afección a la seguridad estructural, en su caso) y se han identificado las diferencias con los procedimientos de hoy. También ha sido necesario adaptar la notación y terminología de la época a los criterios actuales, lo que ha supuesto una dificultad añadida. -Desarrollo del documento: A partir del estudio previo se han ido desarrollando los siguientes documentos, que conforman el contenido de la tesis: o Personajes e instituciones relevantes por sus aportaciones al conocimiento de las estructuras de hormigón (investigación, normativa, docencia). o Caracterización de las propiedades mecánicas de los materiales (hormigón y armaduras), en relación a sus resistencias, diagramas tensión-deformación, módulos de deformación, diagramas momento-curvatura, etc. Se incluye aquí la caracterización clásica de los hormigones, la geometría y naturaleza de las armaduras, etc. o Formatos de seguridad: Se trata de un complejo capítulo del que se pretende extraer la información suficiente que permita a los técnicos de hoy entender los criterios utilizados entonces y compararlos con los actuales. o Estudio de secciones y piezas sometidas a tensiones normales y tangenciales: Se trata de presentar la evolución en el tratamiento de la flexión simple y compuesta, del cortante, del rasante, torsión, etc. Se tratan también en esta parte del estudio aspectos que, no siendo de preocupación directa de los técnicos de antaño (fisuración y deformaciones), tienen hoy mayor importancia frente a cambios de usos y condiciones de durabilidad. o Detalles de armado: Incluye el tratamiento de la adherencia, el anclaje, el solapo de barras, el corte de barras, las disposiciones de armado en función de la geometría de las piezas y sus solicitaciones, etc. Es un capítulo de importancia obvia para los técnicos de hoy. Se incluye un anejo con las referencias más significativas a los estudios experimentales en que se basaron las propuestas que han marcado hito en la evolución del conocimiento. Finalmente, junto a las conclusiones más importantes, se enuncian las propuestas de estudios futuros. This thesis analyzes the criteria with which structures of reinforced concrete have been designed and constructed prior to 1973. Initially, the year 1970 was chosen as starting point, coinciding with the CEB recommendations, but with the development of the thesis it was decided that 1973 was the better option, coinciding with the Spanish regulations of 1973, whose content, format and description introduced the current criteria. The studied period includes the Classic Theory. The intended goals of this thesis are: 1) To cover a clear gap in the study of evolution of knowledge about reinforced concrete. The concept and accomplishments achieved by reinforced concrete itself has been treated in a very complete way by the main researchers in this area, but not the evolution of knowledge in this subject area. 2) To help the engineers understand structural configurations, geometries, dispositions of steel, safety formats etc, that will serve as preliminary judgments by experts on existing structures. To be a reference to the existing studies about the valuation of resistant capacity of existing constructions, constituting a basic study of a pre-regulation document. This thesis intends to be a help for the current generation of engineers who need to preserve and repair reinforced concrete structures that have existed for a significant number of years. Most of these structures in question were constructed more than 40 years ago, and it is necessary to know the criteria that influenced their design, the calculation and the construction. This thesis intends to determine the safety limits of the old structures and analyze them in the context of the current regulations and their methodology. Thus, it will then be possible to determine the safety of these structures, after being measured and calculated with the current criteria. This will allow the engineers to optimize the treatment of such a structure. This work considers the evolution of the knowledge, so constructive methods are not included. Related to the design criteria, there existed until middle of the 20th century a large number of diverse European tests and regulations, such as the Prussian norm of 1904, the Circular French Order of 1906, the Congress of Liège of 1930, as well as individual engineers’ own notes and criteria which incorporated the results of their own tests. From the second half of the 20th century, the contributions of Spanish engineers as Alfredo Páez Balaca, Eduardo Torroja and Pedro Jiménez Montoya, among others, were significant and this allowed the advancement of the criteria of the calculation of safety standards of concrete structures, many of which still exist to the present day. The design and calculation of reinforced concrete structures by the Classic Theory, was based on the ‘Critical Bending Moment’, when concrete and steel achieve their admissible tensions, that allows the best employment of materials and the best ductility. If the bending moment is major than the critical bending moment, will be necessary to introduce compression steel. After the study of the designs of many existing structures of that time by the author of this thesis, including the Historical Collections of Juan Manuel de Zafra, Eugenio Ribera and Carlos Fernandez Casado, the conclusion is that the geometric definition of the structures does not correspond exactly with the critical bending moment inherent in the structures. The parameters of these calculations changed throughout the years. The principal reason that can be outlined is that the materials were improving gradually and the number of calculated uncertainties were decreasing, thus allowing the reduction of the safety coefficients to use in the calculation. For example, concrete used a coefficient of 4 towards the end of the 19th century, which evolved to 3,57 after the publication of the Circular French Order of 1906, and then to 3 after the Spanish Instruction of 1939. In the case of the steel, a much more consistent material, the safety coefficient remained almost constant throughout the years, with a value of 2. Other reasons related to the evolution of the calculation parameters were that the tests and research undertaken by an ever-increasing number of engineers then allowed a more complete knowledge of the behavior of reinforced concrete. What is surprising is the extent of knowledge that existed about the behavior of the concrete from the outset. Engineers from the early years knew that the behavior of the concrete was non-linear, but they limited the work to a linear tension-deformation range. This was due to the difficulties of work in a non-linear range, because they did not have laboratories to test concrete, or facilities such as computers with appropriate software, something unthinkable today. These were the main reasons engineers of previous generations limited the tensions and deformations of a particular material to a known range. The modus operandi followed for the development of this thesis is the following one: -Document study: engineers’ documents, recommendations and regulations generated in this area, both from Spain or overseas, have been studied in a systematic way in accordance with the index of the document. In this process, a lack of knowledge has been detected concerning structural safety, and differences to current procedures have been identified and noted. Also, it has been necessary to adapt the notation and terminology of the Classic Theory to the current criteria, which has imposed an additional difficulty. -Development of the thesis: starting from the basic study, the next chapters of this thesis have been developed and expounded upon: o People and relevant institutions for their contribution to the knowledge about reinforced concrete structures (investigation, regulation, teaching). Determination of the mechanical properties of the materials (concrete and steel), in relation to their resistances, tension-deformation diagrams, modules of deformation, moment-curvature diagrams, etc. Included are the classic characterizations of concrete, the geometry and nature of the steel, etc. Safety formats: this is a very difficult chapter from which it is intended to provide enough information that will then allow the present day engineer to understand the criteria used in the Classic Theory and then to compare them with the current theories. Study of sections and pieces subjected to normal and tangential tensions: it intends to demonstrate the evolution in the treatment of the simple and complex flexion, shear, etc. Other aspects examined include aspects that were not very important in the Classic Theory but currently are, such as deformation and fissures. o Details of reinforcement: it includes the treatment of the adherence, the anchorage, the lapel of bars, the cut of bars, the dispositions of reinforcement depending on the geometry of the pieces and the solicitations, etc. It is a chapter of obvious importance for current engineers. The document will include an annex with the most references to the most significant experimental studies on which were based the proposals that have become a milestone in the evolution of knowledge in this area. Finally, there will be included conclusions and suggestions of future studies. A deep study of the documentation and researchers of that time has been done, juxtaposing their criteria and results with those considered relevant today, and giving a comparison between the resultant safety standards according to the Classic Theory criteria and currently used criteria. This thesis fundamentally intends to be a guide for engineers who have to treat or repair a structure constructed according to the Classic Theory criteria.

Límites de ductilidad y de servicio al cálculo en carga última de estructuras de hormigón armado y de fábrica

En este trabajo se aborda una cuestión central en el diseño en carga última de estructuras de hormigón armado y de fábrica: la posibilidad efectiva de que las deformaciones plásticas necesarias para verificar un estado de rotura puedan ser alcanzadas por las regiones de la estructura que deban desarrollar su capacidad última para verificar tal estado. Así, se parte de las decisiones de diseño que mediante mera estática aseguran un equilibrio de la estructura para las cargas últimas que deba resistir, pero determinando directamente el valor de las deformaciones necesarias para llegar a tal estado. Por tanto, no se acude a los teoremas de rotura sin más, sino que se formula el problema desde un punto de vista elastoplástico. Es decir, no se obvia el recorrido que la estructura deba realizar en un proceso de carga incremental monótono, de modo que las regiones no plastificadas contribuyen a coaccionar las libres deformaciones plásticas que, en la teoría de rotura, se suponen. En términos de trabajo y energía, se introduce en el balance del trabajo de las fuerzas externas y en el de la energía de deformación, aquella parte del sistema que no ha plastificado. Establecido así el balance energético como potencial del sistema es cuando la condición de estacionariedad del mismo hace determinados los campos de desplazamientos y, por tanto, el de las deformaciones plásticas también. En definitiva, se trata de un modo de verificar si la ductilidad de los diseños previstos es suficiente, y en qué medida, para verificar el estado de rotura previsto, para unas determinadas cargas impuestas. Dentro del desarrollo teórico del problema, se encuentran ciertas precisiones importantes. Entre ellas, la verificación de que el estado de rotura a que se llega de manera determinada mediante el balance energético elasto-plástico satisface las condiciones de la solución de rotura que los teoremas de carga última predicen, asegurando, por tanto, que la solución determinada -unicidad del problema elásticocoincide con el teorema de unicidad de la carga de rotura, acotando además cuál es el sistema de equilibrio y cuál es la deformada de colapso, aspectos que los teoremas de rotura no pueden asegurar, sino sólo el valor de la carga última a verificar. Otra precisión se basa en la particularidad de los casos en que el sistema presenta una superficie de rotura plana, haciendo infinitas las posibilidades de equilibrio para una misma deformada de colapso determinada, lo que está en la base de, aparentemente, poder plastificar a antojo en vigas y arcos. Desde el planteamiento anterior, se encuentra entonces que existe una condición inherente a cualquier sistema, definidas unas leyes constitutivas internas, que permite al mismo llegar al inicio del estado de rotura sin demandar deformación plástica alguna, produciéndose la plastificación simultánea de todas las regiones que hayan llegado a su solicitación de rotura. En cierto modo, se daría un colapso de apariencia frágil. En tal caso, el sistema conserva plenamente hasta el final su capacidad dúctil y tal estado actúa como representante canónico de cualquier otra solución de equilibrio que con idéntico criterio de diseño interno se prevea para tal estructura. En la medida que el diseño se acerque o aleje de la solución canónica, la demanda de ductilidad del sistema para verificar la carga última será menor o mayor. Las soluciones que se aparten en exceso de la solución canónica, no verificarán el estado de rotura previsto por falta de ductilidad: la demanda de deformación plástica de alguna región plastificada estará más allá de la capacidad de la misma, revelándose una carga de rotura por falta de ductilidad menor que la que se preveía por mero equilibrio. Para la determinación de las deformaciones plásticas de las rótulas, se ha tomado un modelo formulado mediante el Método de los Elementos de Contorno, que proporciona un campo continuo de desplazamientos -y, por ende, de deformaciones y de tensiones- incluso en presencia de fisuras en el contorno. Importante cuestión es que se formula la diferencia, nada desdeñable, de la capacidad de rotación plástica de las secciones de hormigón armado en presencia de cortante y en su ausencia. Para las rótulas de fábrica, la diferencia se establece para las condiciones de la excentricidad -asociadas al valor relativo de la compresión-, donde las diferencias entres las regiones plastificadas con esfuerzo normal relativo alto o bajo son reseñables. Por otro lado, si bien de manera un tanto secundaria, las condiciones de servicio también imponen un límite al diseño previo en carga última deseado. La plastificación lleva asociadas deformaciones considerables, sean locales como globales. Tal cosa impone que, en estado de servicio, si la plastificación de alguna región lleva asociadas fisuraciones excesivas para el ambiente del entorno, la solución sea inviable por ello. Asimismo, las deformaciones de las estructuras suponen un límite severo a las posibilidades de su diseño. Especialmente en edificación, las deformaciones activas son un factor crítico a la hora de decidirse por una u otra solución. Por tanto, al límite que se impone por razón de ductilidad, se debe añadir el que se imponga por razón de las condiciones de servicio. Del modo anterior, considerando las condiciones de ductilidad y de servicio en cada caso, se puede tasar cada decisión de diseño con la previsión de cuáles serán las consecuencias en su estado de carga última y de servicio. Es decir, conocidos los límites, podemos acotar cuáles son los diseños a priori que podrán satisfacer seguro las condiciones de ductilidad y de servicio previstas, y en qué medida. Y, en caso de no poderse satisfacer, qué correcciones debieran realizarse sobre el diseño previo para poderlas cumplir. Por último, de las consecuencias que se extraen de lo estudiado, se proponen ciertas líneas de estudio y de experimentación para poder llegar a completar o expandir de manera práctica los resultados obtenidos. ABSTRACT This work deals with a main issue for the ultimate load design in reinforced concrete and masonry structures: the actual possibility that needed yield strains to reach a ultimate state could be reached by yielded regions on the structure that should develop their ultimate capacity to fulfill such a state. Thus, some statically determined design decisions are posed as a start for prescribed ultimate loads to be counteracted, but finding out the determined value of the strains needed to reach the ultimate load state. Therefore, ultimate load theorems are not taken as they are, but a full elasto-plastic formulation point of view is used. As a result, the path the structure must develop in a monotonus increasing loading procedure is not neglected, leading to the fact that non yielded regions will restrict the supposed totally free yield strains under a pure ultimate load theory. In work and energy terms, in the overall account of external forces work and internal strain energy, those domains in the body not reaching their ultimate state are considered. Once thus established the energy balance of the system as its potential, by imposing on it the stationary condition, both displacements and yield strains appear as determined values. Consequently, what proposed is a means for verifying whether the ductility of prescribed designs is enough and the extent to which they are so, for known imposed loads. On the way for the theoretical development of the proposal, some important aspects have been found. Among these, the verification that the conditions for the ultimate state reached under the elastoplastic energy balance fulfills the conditions prescribed for the ultimate load state predicted through the ultimate load theorems, assuring, therefore, that the determinate solution -unicity of the elastic problemcoincides with the unicity ultimate load theorem, determining as well which equilibrium system and which collapse shape are linked to it, being these two last aspects unaffordable by the ultimate load theorems, that make sure only which is the value of the ultimate load leading to collapse. Another aspect is based on the particular case in which the yield surface of the system is flat -i.e. expressed under a linear expression-, turning out infinite the equilibrium possibilities for one determined collapse shape, which is the basis of, apparently, deciding at own free will the yield distribution in beams and arches. From the foresaid approach, is then found that there is an inherent condition in any system, once defined internal constitutive laws, which allows it arrive at the beginning of the ultimate state or collapse without any yield strain demand, reaching the collapse simultaneously for all regions that have come to their ultimate strength. In a certain way, it would appear to be a fragile collapse. In such a case case, the system fully keeps until the end its ductility, and such a state acts as a canonical representative of any other statically determined solution having the same internal design criteria that could be posed for the that same structure. The extent to which a design is closer to or farther from the canonical solution, the ductility demand of the system to verify the ultimate load will be higher or lower. The solutions being far in excess from the canonical solution, will not verify the ultimate state due to lack of ductility: the demand for yield strains of any yielded region will be beyond its capacity, and a shortcoming ultimate load by lack of ductility will appear, lower than the expected by mere equilibrium. For determining the yield strains of plastic hinges, a Boundary Element Method based model has been used, leading to a continuous displacement field -therefore, for strains and stresses as well- even if cracks on the boundary are present. An important aspect is that a remarkable difference is found in the rotation capacity between plastic hinges in reinforced concrete with or without shear. For masonry hinges, such difference appears when dealing with the eccentricity of axial forces -related to their relative value of compression- on the section, where differences between yield regions under high or low relative compressions are remarkable. On the other hand, although in a certain secondary manner, serviceability conditions impose limits to the previous ultimate load stated wanted too. Yield means always big strains and deformations, locally and globally. Such a thing imposes, for serviceability states, that if a yielded region is associated with too large cracking for the environmental conditions, the predicted design will be unsuitable due to this. Furthermore, displacements must be restricted under certain severe limits that restrain the possibilities for a free design. Especially in building structures, active displacements are a critical factor when chosing one or another solution. Then, to the limits due to ductility reasons, other limits dealing with serviceability conditions shoud be added. In the foresaid way, both considering ductility and serviceability conditions in every case, the results for ultimate load and serviceability to which every design decision will lead can be bounded. This means that, once the limits are known, it is possible to bound which a priori designs will fulfill for sure the prescribed ductility and serviceability conditions, and the extent to wich they will be fulfilled, And, in case they were not, which corrections must be performed in the previous design so that it will. Finally, from the consequences derived through what studied, several study and experimental fields are proposed, in order to achieve a completeness and practical expansion of the obtained results.

Coherent structures in statistically-stationary homogeneous shear turbulence

La región cerca de la pared de flujos turbulentos de pared ya está bien conocido debido a su bajo número de Reynolds local y la separación escala estrecha. La región lejos de la pared (capa externa) no es tan interesante tampoco, ya que las estadísticas allí se escalan bien por las unidades exteriores. La región intermedia (capa logarítmica), sin embargo, ha estado recibiendo cada vez más atención debido a su propiedad auto-similares. Además, de acuerdo a Flores et al. (2007) y Flores & Jiménez (2010), la capa logarítmica es más o menos independiente de otras capas, lo que implica que podría ser inspeccionado mediante el aislamiento de otras dos capas, lo que reduciría significativamente los costes computacionales para la simulación de flujos turbulentos de pared. Algunos intentos se trataron después por Mizuno & Jiménez (2013), quien simulan la capa logarítmica sin la región cerca de la pared con estadísticas obtenidas de acuerdo razonablemente bien con los de las simulaciones completas. Lo que más, la capa logarítmica podría ser imitado por otra turbulencia sencillo de cizallamiento de motor. Por ejemplo, Pumir (1996) encontró que la turbulencia de cizallamiento homogéneo estadísticamente estacionario (SS-HST) también irrumpe, de una manera muy similar al proceso de auto-sostenible en flujos turbulentos de pared. Según los consideraciones arriba, esta tesis trata de desvelar en qué medida es la capa logarítmica de canales similares a la turbulencia de cizalla más sencillo, SS-HST, mediante la comparación de ambos cinemática y la dinámica de las estructuras coherentes en los dos flujos. Resultados sobre el canal se muestran mediante Lozano-Durán et al. (2012) y Lozano-Durán & Jiménez (2014b). La hoja de ruta de esta tarea se divide en tres etapas. En primer lugar, SS-HST es investigada por medio de un código nuevo de simulación numérica directa, espectral en las dos direcciones horizontales y compacto-diferencias finitas en la dirección de la cizalla. Sin utiliza remallado para imponer la condición de borde cortante periódica. La influencia de la geometría de la caja computacional se explora. Ya que el HST no tiene ninguna longitud característica externa y tiende a llenar el dominio computacional, las simulaciopnes a largo plazo del HST son ’mínimos’ en el sentido de que contiene sólo unas pocas estructuras media a gran escala. Se ha encontrado que el límite principal es el ancho de la caja de la envergadura, Lz, que establece las escalas de longitud y velocidad de la turbulencia, y que las otras dos dimensiones de la caja debe ser suficientemente grande (Lx > 2LZ, Ly > Lz) para evitar que otras direcciones estando limitado también. También se encontró que las cajas de gran longitud, Lx > 2Ly, par con el paso del tiempo la condición de borde cortante periódica, y desarrollar fuertes ráfagas linealizadas no físicos. Dentro de estos límites, el flujo muestra similitudes y diferencias interesantes con otros flujos de cizalla, y, en particular, con la capa logarítmica de flujos turbulentos de pared. Ellos son exploradas con cierto detalle. Incluyen un proceso autosostenido de rayas a gran escala y con una explosión cuasi-periódica. La escala de tiempo de ruptura es de aproximadamente universales, ~20S~l(S es la velocidad de cizallamiento media), y la disponibilidad de dos sistemas de ruptura diferentes permite el crecimiento de las ráfagas a estar relacionado con algo de confianza a la cizalladura de turbulencia inicialmente isotrópico. Se concluye que la SS-HST, llevado a cabo dentro de los parámetros de cílculo apropiados, es un sistema muy prometedor para estudiar la turbulencia de cizallamiento en general. En segundo lugar, las mismas estructuras coherentes como en los canales estudiados por Lozano-Durán et al. (2012), es decir, grupos de vórticidad (fuerte disipación) y Qs (fuerte tensión de Reynolds tangencial, -uv) tridimensionales, se estudia mediante simulación numérica directa de SS-HST con relaciones de aspecto de cuadro aceptables y número de Reynolds hasta Rex ~ 250 (basado en Taylor-microescala). Se discute la influencia de la intermitencia de umbral independiente del tiempo. Estas estructuras tienen alargamientos similares en la dirección sentido de la corriente a las familias separadas en los canales hasta que son de tamaño comparable a la caja. Sus dimensiones fractales, longitudes interior y exterior como una función del volumen concuerdan bien con sus homólogos de canales. El estudio sobre sus organizaciones espaciales encontró que Qs del mismo tipo están alineados aproximadamente en la dirección del vector de velocidad en el cuadrante al que pertenecen, mientras Qs de diferentes tipos están restringidos por el hecho de que no debe haber ningún choque de velocidad, lo que hace Q2s (eyecciones, u < 0,v > 0) y Q4s (sweeps, u > 0,v < 0) emparejado en la dirección de la envergadura. Esto se verifica mediante la inspección de estructuras de velocidad, otros cuadrantes como la uw y vw en SS-HST y las familias separadas en el canal. La alineación sentido de la corriente de Qs ligada a la pared con el mismo tipo en los canales se debe a la modulación de la pared. El campo de flujo medio condicionado a pares Q2-Q4 encontró que los grupos de vórticidad están en el medio de los dos, pero prefieren los dos cizalla capas alojamiento en la parte superior e inferior de Q2s y Q4s respectivamente, lo que hace que la vorticidad envergadura dentro de las grupos de vórticidad hace no cancele. La pared amplifica la diferencia entre los tamaños de baja- y alta-velocidad rayas asociados con parejas de Q2-Q4 se adjuntan como los pares alcanzan cerca de la pared, el cual es verificado por la correlación de la velocidad del sentido de la corriente condicionado a Q2s adjuntos y Q4s con diferentes alturas. Grupos de vórticidad en SS-HST asociados con Q2s o Q4s también están flanqueadas por un contador de rotación de los vórtices sentido de la corriente en la dirección de la envergadura como en el canal. La larga ’despertar’ cónica se origina a partir de los altos grupos de vórticidad ligada a la pared han encontrado los del Álamo et al. (2006) y Flores et al. (2007), que desaparece en SS-HST, sólo es cierto para altos grupos de vórticidad ligada a la pared asociados con Q2s pero no para aquellos asociados con Q4s, cuyo campo de flujo promedio es en realidad muy similar a la de SS-HST. En tercer lugar, las evoluciones temporales de Qs y grupos de vórticidad se estudian mediante el uso de la método inventado por Lozano-Durán & Jiménez (2014b). Las estructuras se clasifican en las ramas, que se organizan más en los gráficos. Ambas resoluciones espaciales y temporales se eligen para ser capaz de capturar el longitud y el tiempo de Kolmogorov puntual más probable en el momento más extrema. Debido al efecto caja mínima, sólo hay un gráfico principal consiste en casi todas las ramas, con su volumen y el número de estructuras instantáneo seguien la energía cinética y enstrofía intermitente. La vida de las ramas, lo que tiene más sentido para las ramas primarias, pierde su significado en el SS-HST debido a las aportaciones de ramas primarias al total de Reynolds estrés o enstrofía son casi insignificantes. Esto también es cierto en la capa exterior de los canales. En cambio, la vida de los gráficos en los canales se compara con el tiempo de ruptura en SS-HST. Grupos de vórticidad están asociados con casi el mismo cuadrante en términos de sus velocidades medias durante su tiempo de vida, especialmente para los relacionados con las eyecciones y sweeps. Al igual que en los canales, las eyecciones de SS-HST se mueven hacia arriba con una velocidad promedio vertical uT (velocidad de fricción) mientras que lo contrario es cierto para los barridos. Grupos de vórticidad, por otra parte, son casi inmóvil en la dirección vertical. En la dirección de sentido de la corriente, que están advección por la velocidad media local y por lo tanto deforman por la diferencia de velocidad media. Sweeps y eyecciones se mueven más rápido y más lento que la velocidad media, respectivamente, tanto por 1.5uT. Grupos de vórticidad se mueven con la misma velocidad que la velocidad media. Se verifica que las estructuras incoherentes cerca de la pared se debe a la pared en vez de pequeño tamaño. Los resultados sugieren fuertemente que las estructuras coherentes en canales no son especialmente asociado con la pared, o incluso con un perfil de cizalladura dado. ABSTRACT Since the wall-bounded turbulence was first recognized more than one century ago, its near wall region (buffer layer) has been studied extensively and becomes relatively well understood due to the low local Reynolds number and narrow scale separation. The region just above the buffer layer, i.e., the logarithmic layer, is receiving increasingly more attention nowadays due to its self-similar property. Flores et al. (20076) and Flores & Jim´enez (2010) show that the statistics of logarithmic layer is kind of independent of other layers, implying that it might be possible to study it separately, which would reduce significantly the computational costs for simulations of the logarithmic layer. Some attempts were tried later by Mizuno & Jimenez (2013), who simulated the logarithmic layer without the buffer layer with obtained statistics agree reasonably well with those of full simulations. Besides, the logarithmic layer might be mimicked by other simpler sheardriven turbulence. For example, Pumir (1996) found that the statistically-stationary homogeneous shear turbulence (SS-HST) also bursts, in a manner strikingly similar to the self-sustaining process in wall-bounded turbulence. Based on these considerations, this thesis tries to reveal to what extent is the logarithmic layer of channels similar to the simplest shear-driven turbulence, SS-HST, by comparing both kinematics and dynamics of coherent structures in the two flows. Results about the channel are shown by Lozano-Dur´an et al. (2012) and Lozano-Dur´an & Jim´enez (20146). The roadmap of this task is divided into three stages. First, SS-HST is investigated by means of a new direct numerical simulation code, spectral in the two horizontal directions and compact-finite-differences in the direction of the shear. No remeshing is used to impose the shear-periodic boundary condition. The influence of the geometry of the computational box is explored. Since HST has no characteristic outer length scale and tends to fill the computational domain, longterm simulations of HST are ‘minimal’ in the sense of containing on average only a few large-scale structures. It is found that the main limit is the spanwise box width, Lz, which sets the length and velocity scales of the turbulence, and that the two other box dimensions should be sufficiently large (Lx > 2LZ, Ly > Lz) to prevent other directions to be constrained as well. It is also found that very long boxes, Lx > 2Ly, couple with the passing period of the shear-periodic boundary condition, and develop strong unphysical linearized bursts. Within those limits, the flow shows interesting similarities and differences with other shear flows, and in particular with the logarithmic layer of wallbounded turbulence. They are explored in some detail. They include a self-sustaining process for large-scale streaks and quasi-periodic bursting. The bursting time scale is approximately universal, ~ 20S~l (S is the mean shear rate), and the availability of two different bursting systems allows the growth of the bursts to be related with some confidence to the shearing of initially isotropic turbulence. It is concluded that SS-HST, conducted within the proper computational parameters, is a very promising system to study shear turbulence in general. Second, the same coherent structures as in channels studied by Lozano-Dur´an et al. (2012), namely three-dimensional vortex clusters (strong dissipation) and Qs (strong tangential Reynolds stress, -uv), are studied by direct numerical simulation of SS-HST with acceptable box aspect ratios and Reynolds number up to Rex ~ 250 (based on Taylor-microscale). The influence of the intermittency to time-independent threshold is discussed. These structures have similar elongations in the streamwise direction to detached families in channels until they are of comparable size to the box. Their fractal dimensions, inner and outer lengths as a function of volume agree well with their counterparts in channels. The study about their spatial organizations found that Qs of the same type are aligned roughly in the direction of the velocity vector in the quadrant they belong to, while Qs of different types are restricted by the fact that there should be no velocity clash, which makes Q2s (ejections, u < 0, v > 0) and Q4s (sweeps, u > 0, v < 0) paired in the spanwise direction. This is verified by inspecting velocity structures, other quadrants such as u-w and v-w in SS-HST and also detached families in the channel. The streamwise alignment of attached Qs with the same type in channels is due to the modulation of the wall. The average flow field conditioned to Q2-Q4 pairs found that vortex clusters are in the middle of the pair, but prefer to the two shear layers lodging at the top and bottom of Q2s and Q4s respectively, which makes the spanwise vorticity inside vortex clusters does not cancel. The wall amplifies the difference between the sizes of low- and high-speed streaks associated with attached Q2-Q4 pairs as the pairs reach closer to the wall, which is verified by the correlation of streamwise velocity conditioned to attached Q2s and Q4s with different heights. Vortex clusters in SS-HST associated with Q2s or Q4s are also flanked by a counter rotating streamwise vortices in the spanwise direction as in the channel. The long conical ‘wake’ originates from tall attached vortex clusters found by del A´ lamo et al. (2006) and Flores et al. (2007b), which disappears in SS-HST, is only true for tall attached vortices associated with Q2s but not for those associated with Q4s, whose averaged flow field is actually quite similar to that in SS-HST. Third, the temporal evolutions of Qs and vortex clusters are studied by using the method invented by Lozano-Dur´an & Jim´enez (2014b). Structures are sorted into branches, which are further organized into graphs. Both spatial and temporal resolutions are chosen to be able to capture the most probable pointwise Kolmogorov length and time at the most extreme moment. Due to the minimal box effect, there is only one main graph consist by almost all the branches, with its instantaneous volume and number of structures follow the intermittent kinetic energy and enstrophy. The lifetime of branches, which makes more sense for primary branches, loses its meaning in SS-HST because the contributions of primary branches to total Reynolds stress or enstrophy are almost negligible. This is also true in the outer layer of channels. Instead, the lifetime of graphs in channels are compared with the bursting time in SS-HST. Vortex clusters are associated with almost the same quadrant in terms of their mean velocities during their life time, especially for those related with ejections and sweeps. As in channels, ejections in SS-HST move upwards with an average vertical velocity uτ (friction velocity) while the opposite is true for sweeps. Vortex clusters, on the other hand, are almost still in the vertical direction. In the streamwise direction, they are advected by the local mean velocity and thus deformed by the mean velocity difference. Sweeps and ejections move faster and slower than the mean velocity respectively, both by 1.5uτ . Vortex clusters move with the same speed as the mean velocity. It is verified that the incoherent structures near the wall is due to the wall instead of small size. The results suggest that coherent structures in channels are not particularly associated with the wall, or even with a given shear profile.

La introducción del hormigón armado en España: razón constructiva de su evolución

España se incorporó a la técnica del hormigón armado con más de dos décadas de retraso respecto a Francia o Alemania. En 1890, en Europa se construían ya estructuras de hormigón armado de cierta envergadura y complejidad. En España hubo que esperar hasta 1893 para la primera obra en hormigón armado, que fue un sencillo depósito descubierto en Puigverd (Lérida), ejecutado por el ingeniero militar Francesc Macià con patente Monier. En 1898, de la mano de Hennebique, se empezó la construcción de los dos primeros edificios con estructura de hormigón armado en España. Fueron dos obras puntuales, con proyectos importados de Francia, pero necesarias para introducir de manera definitiva el material. En paralelo, en París, se estaban edificando en hormigón armado la mayoría de los pabellones de la Exposición Universal de 1900. En el cambio de siglo, las construcciones de hormigón armado habían alcanzado ya la madurez proyectual y técnica en Europa. A pesar de la incorporación tardía, se puede constatar por las obras ejecutadas que en un periodo corto de tiempo, entre 1901 y 1906, se alcanzó en España prácticamente el mismo nivel técnico y constructivo que tenían el resto de los países que fueron pioneros en el empleo del hormigón armado. El desarrollo e implantación de una técnica constructiva no es un proceso lineal, y son muchos los factores que intervienen. Las patentes tuvieron una gran importancia en el desarrollo inicial del hormigón armado. Estas ofrecían un producto que funcionaba. Las primeras estructuras de hormigón armado no se calculaban y se construían siguiendo una reglamentación, se compraban. Y el resultado de esa “compra” solía ser, en la mayoría de los casos, satisfactorio. Las patentes vendían sistemas estructurales cuyo funcionamiento estaba corroborado por la experiencia y la pericia de su inventor. Esta investigación parte de la hipótesis de que las patentes sobre cemento y hormigón armado depositadas en España entre 1884 y 1906 fueron uno de los factores que proporcionaron a los técnicos y a las empresas españolas una pericia constructiva sólida en el empleo del hormigón armado. En este trabajo se aborda el estudio del proceso de introducción del hormigón armado en España desde una perspectiva fundamentalmente técnica, incorporando las patentes como una de las razones constructivas que explican su rápida evolución y generalización en un periodo de tiempo breve: 1901-1906. En este proceso se contextualiza y analiza una de las figuras que se considera fundamental en los primeros años del hormigón armado en España, la del ingeniero Juan Manuel de Zafra y Estevan. Esta tesis analiza las patentes de hormigón armado desde el punto de vista estadístico y constructivo. Desde ambas perspectivas se verifica la hipótesis de partida de esta investigación, concluyendo que las patentes fueron una de las razones constructivas de la evolución del hormigón armado en España y de su rápida implantación. ABSTRACT Spain incorporated the reinforced concrete technique more than two decades after France and Germany. In central Europe reinforced concrete structures of considerable size and complexity were being built in 1890, while in Spain it was not until 1893 that the first work, a simple open air water tank, was implemented in Puigverd (Lleida) by the military engineer Francesc Macià with a Monier patent. In 1898 the construction of the first two buildings with reinforced concrete structure in Spain started, with the guidance by Hennebique. They were two isolated cases with projects imported from France, but playing a key role to definitively introduce the material in Spain. In parallel, in Paris, most of the pavilions of the 1900 World Expo were being built in reinforced concrete. At the turn of the century reinforced concrete buildings had reached maturity both as a technology and as a design practice. Despite the late assumption of the material, the works carried out in the very short period between 1901 and 1906 clearly show that Spain reached practically the same technical and constructive level as the other pioneering countries in the use of reinforced concrete. The development and implementation of a constructive technique is never a linear process, there are many factors involved. The patents offered a successful product. Initial reinforced concrete structures were not calculated and built according to regulations, they were bought. And this purchase in most cases was satisfactory for the required use. Patents sold structural systems whose performance was supported by the experience and expertise of its inventor. The hypothesis of this research is based upon the assumption that the cement and concrete patents registered in Spain between 1884 and 1906 were one of the factors that provided Spanish technicians and companies with a solid constructive expertise in the use of reinforced concrete. This investigation studies the introduction of reinforced concrete to Spain from a predominantly technical perspective, incorporating patents as the constructive reason for the rapid evolution and spread in such a short period of time: 1901-1906. Along the way, the role of engineer J. M. de Zafra, generally considered a key agent in the initial years of reinforced concrete in Spain, is contextualized and analyzed. This dissertation analyzes the patents of reinforced concrete from a statistical and constructive point of view. From both perspectives the hypothesis of this research is verified, concluding that patents were one of the constructive reasons for the development of reinforced concrete in Spain.

Colocalization of cell division proteins FtsZ and FtsA to cytoskeletal structures in living Escherichia coli cells by using green fluorescent protein

In the current model for bacterial cell division, FtsZ protein forms a ring that marks the division plane, creating a cytoskeletal framework for the subsequent action of other proteins such as FtsA. This putative protein complex ultimately generates the division septum. Herein we report that FtsZ and FtsA proteins tagged with green fluorescent protein (GFP) colocalize to division-site ring-like structures in living bacterial cells in a visible space between the segregated nucleoids. Cells with higher levels of FtsZ–GFP or with FtsA–GFP plus excess wild-type FtsZ were inhibited for cell division and often exhibited bright fluorescent spiral tubules that spanned the length of the filamentous cells. This suggests that FtsZ may switch from a septation-competent localized ring to an unlocalized spiral under some conditions and that FtsA can bind to FtsZ in both conformations. FtsZ–GFP also formed nonproductive but localized aggregates at a higher concentration that could represent FtsZ nucleation sites. The general domain structure of FtsZ–GFP resembles that of tubulin, since the C terminus of FtsZ is not required for polymerization but may regulate polymerization state. The N-terminal portion of Rhizobium FtsZ polymerized in Escherichia coli and appeared to copolymerize with E. coli FtsZ, suggesting a degree of interspecies functional conservation. Analysis of several deletions of FtsA–GFP suggests that multiple segments of FtsA are important for its localization to the FtsZ ring.

A Role for the GSG Domain in Localizing Sam68 to Novel Nuclear Structures in Cancer Cell Lines

The GSG (GRP33, Sam68, GLD-1) domain is a protein module found in an expanding family of RNA-binding proteins. The numerous missense mutations identified genetically in the GSG domain support its physiological role. Although the exact function of the GSG domain is not known, it has been shown to be required for RNA binding and oligomerization. Here it is shown that the Sam68 GSG domain plays a role in protein localization. We show that Sam68 concentrates into novel nuclear structures that are predominantly found in transformed cells. These Sam68 nuclear bodies (SNBs) are distinct from coiled bodies, gems, and promyelocytic nuclear bodies. Electron microscopic studies show that SNBs are distinct structures that are enriched in phosphorus and nitrogen, indicating the presence of nucleic acids. A GFP-Sam68 fusion protein had a similar localization as endogenous Sam68 in HeLa cells, diffusely nuclear with two to five SNBs. Two other GSG proteins, the Sam68-like mammalian proteins SLM-1 and SLM-2, colocalized with endogenous Sam68 in SNBs. Different GSG domain missense mutations were investigated for Sam68 protein localization. Six separate classes of cellular patterns were obtained, including exclusive SNB localization and association with microtubules. These findings demonstrate that the GSG domain is involved in protein localization and define a new compartment for Sam68, SLM-1, and SLM-2 in cancer cell lines.

Function of RNA secondary structures in transcriptional attenuation of the Bacillus subtilis pyr operon

The Bacillus subtilis pyr operon is regulated by exogenous pyrimidines by a transcriptional attenuation mechanism. Transcription in vitro from pyr DNA templates specifying attenuation regions yielded terminated and read-through transcripts of the expected lengths. Addition of the PyrR regulatory protein plus UMP led to greatly increased termination. Synthetic antisense deoxyoligonucleotides were used to probe possible secondary structures in the pyr mRNA that were proposed to play roles in controlling attenuation. Oligonucleotides predicted to disrupt terminator structures suppressed termination, whereas oligonucleotides predicted to disrupt the stem of antiterminator stem-loops strongly promoted termination at the usual termination site. Oligonucleotides that disrupt a previously unrecognized stem-loop structure, called the anti-antiterminator, the formation of which interferes with formation of the downstream antiterminator, suppressed termination. We propose that transcriptional attenuation of the pyr operon is governed by switching between alternative antiterminator versus anti-antiterminator plus terminator structures, and that PyrR acts by UMP-dependent binding to and stabilization of the anti-antiterminator.

Portosystemic shunting and persistent fetal vascular structures in aryl hydrocarbon receptor-deficient mice

A physiological examination of mice harboring a null allele at the aryl hydrocarbon (Ah) locus revealed that the encoded aryl hydrocarbon receptor plays a role in the resolution of fetal vascular structures during development. Although the aryl hydrocarbon receptor is more commonly studied for its role in regulating xenobiotic metabolism and dioxin toxicity, a developmental role of this protein is supported by the observation that Ah null mice display smaller livers, reduced fecundity, and decreased body weights. Upon investigating the liver phenotype, we found that the decrease in liver size is directly related to a reduction in hepatocyte size. We also found that smaller hepatocyte size is the result of massive portosystemic shunting in null animals. Colloidal carbon uptake and microsphere perfusion studies indicated that 56% of portal blood flow bypasses the liver sinusoids. Latex corrosion casts and angiography demonstrated that shunting is consistent with the existence of a patent ductus venosus in adult animals. Importantly, fetal vascular structures were also observed at other sites. Intravital microscopy demonstrated an immature sinusoidal architecture in the liver and persistent hyaloid arteries in the eyes of adult Ah null mice, whereas corrosion casting experiments described aberrations in kidney vascular patterns.

Sir3-dependent assembly of supramolecular chromatin structures in vitro

Baculovirus-expressed recombinant Sir3p (rSir3p) has been purified to near homogeneity, and its binding to naked DNA, mononucleosomes, and nucleosomal arrays has been characterized in vitro. At stoichiometric levels rSir3p interacts with intact nucleosomal arrays, mononucleosomes, and naked DNA, as evidenced by formation of supershifted species on native agarose gels. Proteolytic removal of the core histone tail domains inhibits but does not completely abolish rSir3p binding to nucleosomal arrays. The linker DNA in the supershifted complexes remains freely accessible to restriction endonuclease digestion, suggesting that both the tail domains and nucleosomal DNA contribute to rSir3p–chromatin interactions. Together these data indicate that rSir3p cross-links individual nucleosomal arrays into supramolecular assemblies whose physical properties transcend those of typical 10-nm and 30-nm fibers. Based on these data we hypothesize that Sir3p functions, at least in part, by mediating reorganization of the canonical chromatin fiber into functionally specialized higher order chromosomal domains.

Association of RNase mitochondrial RNA processing enzyme with ribonuclease P in higher ordered structures in the nucleolus: a possible coordinate role in ribosome biogenesis.

RNase mitochondrial RNA processing enzyme (MRP) is a nucleolar ribonucleoprotein particle that participates in 5.8S ribosomal RNA maturation in eukaryotes. This enzyme shares a polypeptide and an RNA structural motif with ribonuclease P (RNase P), a nuclear endoribonuclease originally described in the nucleus that processes RNA transcripts to generate their mature 5' termini. Both enzymes are also located in mitochondria. This report further characterizes the relationship between RNase MRP and RNase P. Antisense affinity selection with biotinylated 2'-O-methyl oligoribonucleotides and glycerol gradient fractionation experiments demonstrated that small subpopulations of RNase MRP and RNase P associate with each other in vivo in macromolecular complex, possibly 60-80S preribosomes. This latter notion was supported by fluorescence in situ hybridization experiments with antisense oligonucleotides that localized that RNA components of RNase MRP and RNase P to the nucleolus and to discrete cytoplasmic structures. These findings suggest that small subpopulations of RNase MRP and RNase P are physically associated, and that both may function in ribosomal RNA maturation or ribosome assembly.

Evidence for nonrandom hydrophobicity structures in protein chains.

The question of whether proteins originate from random sequences of amino acids is addressed. A statistical analysis is performed in terms of blocked and random walk values formed by binary hydrophobic assignments of the amino acids along the protein chains. Theoretical expectations of these variables from random distributions of hydrophobicities are compared with those obtained from functional proteins. The results, which are based upon proteins in the SWISS-PROT data base, convincingly show that the amino acid sequences in proteins differ from what is expected from random sequences in a statistically significant way. By performing Fourier transforms on the random walks, one obtains additional evidence for nonrandomness of the distributions. We have also analyzed results from a synthetic model containing only two amino acid types, hydrophobic and hydrophilic. With reasonable criteria on good folding properties in terms of thermodynamical and kinetic behavior, sequences that fold well are isolated. Performing the same statistical analysis on the sequences that fold well indicates similar deviations from randomness as for the functional proteins. The deviations from randomness can be interpreted as originating from anticorrelations in terms of an Ising spin model for the hydrophobicities. Our results, which differ from some previous investigations using other methods, might have impact on how permissive with respect to sequence specificity protein folding process is-only sequences with nonrandom hydrophobicity distributions fold well. Other distributions give rise to energy landscapes with poor folding properties and hence did not survive the evolution.

Visualization of intermediate and transition-state structures in protein-tyrosine phosphatase catalysis.

Engineering site-specific amino acid substitutions into the protein-tyrosine phosphatase (PTPase) PTP1 and the dual-specific vaccinia H1-related phosphatase (VHR), has kinetically isolated the two chemical steps of the reaction and provided a rare opportunity for examining transition states and directly observing the phosphoenzyme intermediate. Changing serine to alanine in the active-site sequence motif HCXXGXXRS shifted the rate-limiting step from intermediate formation to intermediate hydrolysis. Using phosphorus 31P NMR, the covalent thiol-phosphate intermediate was directly observed during catalytic turnover. The importance of the conserved aspartic acid (D92 in VHR and D181 in PTP1) in both chemical steps was established. Kinetic analysis of D92N and D181N mutants indicated that aspartic acid acts as a general acid by protonating the leaving-group phenolic oxygen. Structure-reactivity experiments with native and aspartate mutant enzymes established that proton transfer is concomitant with P-O cleavage, such that no charge develops on the phenolic oxygen. Steady- and presteady-state kinetics, as well as NMR analysis of the double mutant D92N/S131A (VHR), suggested that the conserved aspartic acid functions as a general base during intermediate hydrolysis. As a general base, aspartate would activate a water molecule to facilitate nucleophilic attack. The amino acids involved in transition-state stabilization for cysteinylphosphate hydrolysis were confirmed by the x-ray structure of the Yersinia PTPase complexed with vanadate, a transition-state mimic that binds covalently to the active-site cysteine. Consistent with the NMR, x-ray, biochemical, and kinetic data, a unifying mechanism for catalysis is proposed.

Core binding factor beta-smooth muscle myosin heavy chain chimeric protein involved in acute myeloid leukemia forms unusual nuclear rod-like structures in transformed NIH 3T3 cells.

Patients with the M4Eo subtype of acute myeloid leukemia almost invariably are found to have an inversion of chromosome 16 in their leukemic cells, which results in a gene fusion between the transcription factor called core binding factor beta (CBFbeta) on 16q and a smooth muscle myosin heavy chain (SMMHC) gene on 16p. Subcellular localizations of the wild-type CBFbeta and the CBFbeta-SMMHC fusion protein were determined by immunofluorescence of NIH 3T3 cells that overexpress wild-type or fusion protein. Normal CBFbeta showed an unexpected perinuclear pattern consistent with primary localization in the Golgi complex. The CBFbeta-SMMHC fusion protein had a very different pattern. Nuclear staining included rod-like crystalline structures as long as 11 microm. The heterodimeric partner of CBFbeta, CBFalpha, formed part of this complex. Cytoplasmic staining included stress fibers that colocalized with actin, probably as a consequence of the myosin heavy chain component of the fusion protein. Deletion of different regions of the CBFbeta portion of the fusion protein showed that binding to CBFalpha was not required for nuclear translocation. However, deletion of parts of the SMMHC domain of the fusion protein involved in myosin-mediated filament formation resulted in proteins that did not form rod-like structures. These observations confirm previous indirect evidence that the CBFbeta-SMMHC fusion protein is capable of forming macromolecular nuclear aggregates and suggests possible models for the mechanism of leukemic transformation.

Paired structures in knowledge representation.

In this position paper we propose a consistent and unifying view to all those basic knowledge representation models that are based on the existence of two somehow opposite fuzzy concepts. A number of these basic models can be found in fuzzy logic and multi-valued logic literature. Here it is claimed that it is the semantic relationship between two paired concepts what determines the emergence of different types of neutrality, namely indeterminacy, ambivalence and conflict, widely used under different frameworks (possibly under different names). It will be shown the potential relevance of paired structures, generated from two paired concepts together with their associated neutrality, all of them to be modeled as fuzzy sets. In this way, paired structures can be viewed as a standard basic model from which different models arise. This unifying view should therefore allow a deeper analysis of the relationships between several existing knowledge representation formalisms, providing a basis from which more expressive models can be later developed.

Avaliação da integridade estrutural de elementos de concreto armado a partir das propriedades modais obtidas por técnicas de excitações aleatórias e transientes

A análise dinâmica experimental tem sido amplamente pesquisada como uma ferramenta de avaliação de integridade de estruturas de concreto armado. Existem técnicas de identificação de danos baseadas em propriedades modais como frequências de ressonâncias, deformadas modais, curvaturas modais e amortecimento. Há também técnicas baseadas na não linearidade da resposta dinâmica, que apesar do grande potencial na detecção de danos, têm sido pouco exploradas nos últimos anos. Este trabalho tem por objetivo avaliar a integridade estrutural de vigas de concreto armado através do comportamento da resposta dinâmica. Foram realizados ensaios dinâmicos em duas vigas de concreto armado com 3,5 m de comprimento, 25 cm de largura, 35 cm de altura e idênticas taxas de armaduras, mas configuradas com barras de aço de diferentes diâmetros, 2 ϕ 16 mm e 8 ϕ 8 mm, respectivamente. Tais vigas, inicialmente íntegras, foram submetidas a ciclos de carregamento e descarregamento com intensidades crescentes até atingir a ruptura do elemento. Após cada ciclo, as propriedades dinâmicas foram avaliadas experimentalmente, com o emprego de técnicas de excitação por sinais do tipo aleatório e tipo transiente, respectivamente, visando determinar parâmetros que indiquem a deterioração gradativa do elemento. Nesses ensaios dinâmicos aplicaram-se diferentes amplitudes da força de excitação. Verificou-se que o aumento da amplitude da força dinâmica de excitação provocou reduções nos valores das frequências de ressonância de 1,1% e 2,4%, associadas, respectivamente, às excitações aleatórias e transientes; e um comportamento não linear dos índices de amortecimento, associados às excitações aleatórias, mantendo um crescimento linear com as excitações transientes. Constatou-se, ainda, que os valores das frequências de ressonância decrescem com a redução de rigidez mecânica, diminuída com o aumento do nível de fissuração induzido nos modelos. Já os valores dos índices de amortecimento, após cada ciclo, se comportaram de forma não linear e assumiram diferentes valores, conforme a técnica de excitação empregada. Acredita-se que esta não linearidade está relacionada aos danos provocados no elemento pela solicitação estrutural e, por consequência, ao processo de como a dissipação de energia é empregada no processo de instauração, configuração e propagação das fissuras nos elementos de concreto armado.