936 resultados para limit cycles
Resumo:
CPV receivers are made of materials with very different lineal expansion coefficients. Strong variations in DNI due to the passage of clouds can cause sudden temperature changes that creates mechanical stress. For common solder and metal filled polymers the plastic limit could be reached causing substantial fatigue. The best forecast of receiver reliability is therefore achieved by applying an intermittent light source with nominal irradiance level and a number of cycles equal to the expected cloud passages for a given site. The UPM has developed specialized equipment, dubbed the LYSS (Light cYcling Stressing Source), for carrying out such experiments. The small thermal capacity of receivers allows simulating more than 25000 cycles per week. The number of deep transients expected for Madrid in 30 years operation, based on available data, is about 45000. We are currently using the system to cycle a ?Ge/Ag Epoxy/aluminum? receiver, which shows no degradation after 20000 cycles. The equipment can cast up to 200 and 70 W/cm2 on 0.1 and 1 cm2 cells, respectively.
Resumo:
After construction of the LYSS (Light cYcling Stressing Source) in early 2014, several CPV receivers, with and without secondary optical element (SOE) have been aged under fast transient illumination cycling,. The test plan for Madrid consisted of 50000 cycles. Receivers with poor heat spreaders showed low reliability but those with thicker metal layers passed the test well. The operation of LYSS along 8 months, after more than 250000 cycles, did not show any significant failure, except lamp reposition every 120 hours, in average. The equipment seems valid for unveiling weak receiver designs with respect to intensive illumination, in steady and transient modes.
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Los adhesivos se conocen y han sido utilizados en multitud de aplicaciones a lo lago de la historia. En la actualidad, la tecnología de la adhesión como método de unión de materiales estructurales está en pleno crecimiento. Los avances científicos han permitido comprender mejor los fenómenos de adhesión, así como, mejorar y desarrollar nuevas formulaciones poliméricas que incrementan el rango de aplicaciones de los adhesivos. Por otro lado, el desarrollo de nuevos materiales y la necesidad de aligerar peso, especialmente en el sector transporte, hace que las uniones adhesivas se introduzcan en aplicaciones hasta ahora reservadas a otros sistemas de unión como la soldadura o las uniones mecánicas, ofreciendo rendimientos similares y, en ocasiones, superiores a los aportados por estas. Las uniones adhesivas ofrecen numerosas ventajas frente a otros sistemas de unión. En la industria aeronáutica y en automoción, las uniones adhesivas logran una reducción en el número de componentes (tales como los tornillos, remaches, abrazaderas) consiguiendo como consecuencia diseños más ligeros y una disminución de los costes de manipulación y almacenamiento, así como una aceleración de los procesos de ensamblaje, y como consecuencia, un aumento de los procesos de producción. En el sector de la construcción y en la fabricación de equipos industriales, se busca la capacidad para soportar la expansión y contracción térmica. Por lo tanto, se usan las uniones adhesivas para evitar producir la distorsión del sustrato al no ser necesario el calentamiento ni la deformación de las piezas cuando se someten a un calentamiento elevado y muy localizado, como en el caso de la soldadura, o cuando se someten a esfuerzos mecánicos localizados, en el caso de montajes remachados. En la industria naval, se están desarrollando técnicas de reparación basadas en la unión adhesiva para distribuir de forma más uniforme y homogénea las tensiones con el objetivo de mejorar el comportamiento frente a fatiga y evitar los problemas asociados a las técnicas de reparación habituales de corte y soldadura. Las uniones adhesivas al no requerir importantes aportes de calor como la soldadura, no producen modificaciones microestructurales indeseables como sucede en la zona fundida o en la zona afectada térmicamente de las uniones soldadas, ni deteriora los recubrimientos protectores de metales de bajo punto de fusión o de naturaleza orgánica. Sin embargo, las uniones adhesivas presentan una desventaja que dificulta su aplicación, se trata de su durabilidad a largo plazo. La primera causa de rotura de los materiales es la rotura por fatiga. Este proceso de fallo es la causa del 85% de las roturas de los materiales estructurales en servicio. La rotura por fatiga se produce cuando se somete al material a la acción de cargas que varían cíclicamente o a vibraciones durante un tiempo prolongado. Las uniones y estructuras sometidas a fatiga pueden fallar a niveles de carga por debajo del límite de resistencia estática del material. La rotura por fatiga en las uniones adhesivas no se produce por un proceso de iniciación y propagación de grieta de forma estable, el proceso de fatiga va debilitando poco a poco la unión hasta que llega un momento que provoca una rotura de forma rápida. Underhill explica este mecanismo como un proceso de daño irreversible de los enlaces más débiles en determinados puntos de la unión. Cuando se ha producido el deterioro de estas zonas más débiles, su área se va incrementando hasta que llega un momento en que la zona dañada es tan amplia que se produce el fallo completo de la unión. En ensayos de crecimiento de grieta realizados sobre probetas preagrietadas en viga con doble voladizo (DCB), Dessureault identifica los procesos de iniciación y crecimiento de grietas en muestras unidas con adhesivo epoxi como una acumulación de microfisuras en la zona próxima al fondo de grieta que, luego, van coalesciendo para configurar la grieta principal. Lo que supone, igualmente, un proceso de daño del adhesivo en la zona de mayor concentración de tensiones que, posteriormente, conduce al fallo de la unión. La presente tesis surge con el propósito de aumentar los conocimientos existentes sobre el comportamiento a fatiga de las uniones adhesivas y especialmente las realizadas con dos tipos de adhesivos estructurales aplicados en aceros con diferentes acabados superficiales. El estudio incluye la obtención de las curvas de tensión frente al número de ciclos hasta el fallo del componente, curvas SN o curvas de Wöhler, que permitirán realizar una estimación de la resistencia a la fatiga de un determinado material o estructura. Los ensayos de fatiga realizados mediante ciclos predeterminados de carga sinusoidales, de amplitud y frecuencia constantes, han permitido caracterizar el comportamiento a la fatiga por el número de ciclos hasta la rotura, siendo el límite de fatiga el valor al que tiende la tensión cuando el número de ciclos es muy grande. En algunos materiales, la fatiga no tiende a un valor límite sino que decrece de forma constante a medida que aumenta el número de ciclos. Para estas situaciones, se ha definido la resistencia a la fatiga (o límite de resistencia) por la tensión en que se produce la rotura para un número de ciclos predeterminado. Todos estos aspectos permitirán un mejor diseño de las uniones y las condiciones de trabajo de los adhesivos con el fin de lograr que la resistencia a fatiga de la unión sea mucho más duradera y el comportamiento total de la unión sea mucho mejor, contribuyendo al crecimiento de la utilización de las uniones adhesivas respecto a otras técnicas. ABSTRACT Adhesives are well-known and have been used in many applications throughout history. At present, adhesion bonding technology of structural materials is experiencing an important growth. Scientific advances have enabled a better understanding of the phenomena of adhesion, as well as to improve and develop new polymeric formulations that increase the range of applications. On the other hand, the development of new materials and the need to save weight, especially in the transport sector, have promote the use of adhesive bonding in many applications previously reserved for other joining technologies such as welded or mechanical joints, presenting similar or even higher performances. Adhesive bonding offers many advantages over other joining methods. For example, in the aeronautic industry and in the automation sector, adhesive bonding allows a reduction in the number of components (such as bolts, rivets, clamps) and as consequence, resulting in lighter designs and a decrease in handling and storage costs, as well as faster assembly processes and an improvement in the production processes. In the construction sector and in the industrial equipment manufacturing, the ability to withstand thermal expansion and contraction is required. Therefore, adhesion bonding technology is used to avoid any distortion of the substrate since this technology does not require heating nor the deformation of the pieces when these are exposed to very high and localized heating, as in welding, or when are subjected to localized mechanical stresses in the case of riveted joints. In the naval industry, repair techniques based in the adhesive bonding are being developed in order to distribute stresses more uniform and homogeneously in order to improve the performance against fatigue and to avoid the problems associated with standard repair techniques as cutting and welding. Adhesive bonding does not require the use of high temperatures and as consequence they do not produce undesirable microstructural changes, as it can be observed in molten zones or in heat-affected zones in the case of welding, neither is there damage of the protective coating of metals with low melting points or polymeric films. However, adhesive bonding presents a disadvantage that limits its application, the low longterm durability. The most common cause of fractures of materials is fatigue fracture. This failure process is the cause of 85% of the fracture of structural materials in service. Fatigue failure occurs when the materials are subjected to the action of cyclic loads or vibrations for a long period of time. The joints and structures subjected to fatigue can fail at stress values below the static strength of the material. Fatigue failure do not occurs by a static and homogeneous process of initiation and propagation of crack. The fatigue process gradually weakens the bond until the moment in which the fracture occurs very rapidly. Underhill explains this mechanism as a process of irreversible damage of the weakest links at certain points of the bonding. When the deterioration in these weaker zones occurs, their area increase until the damage zone is so extensive that the full failure of the joint occurs. During the crack growth tests performed on precracked double-cantilever beam specimen, (DCB), Dessureault identified the processes of crack initiation and growth in samples bonded with epoxy adhesive as a process of accumulation of microcracks on the zone near the crack bottom, then, they coalesced to configure the main crack. This is a damage process of the adhesive in the zone of high stress concentration that leads to failure of the bond. This thesis aims to further the understanding of the fatigue behavior of the adhesive bonding, primarily those based on two different types of structural adhesives used on carbon-steel with different surface treatments. This memory includes the analysis of the SN or Wöhler curves (stress vs. number of cycles curves up to the failure), allowing to carry out an estimation of the fatigue strength of a specific material or structure. The fatigue tests carried out by means of predetermined cycles of sinusoidal loads, with a constant amplitude and frequency, allow the characterisation of the fatigue behaviour. For some materials, there is a maximum stress amplitude below which the material never fails for any number of cycles, known as fatigue limit. In the other hand, for other materials, the fatigue does not tend toward a limit value but decreases constantly as the number of cycles increases. For these situations, the fatigue strength is defined by the stress at which the fracture occurs for a predetermined number of cycles. All these aspects will enable a better joint design and service conditions of adhesives in order to get more durable joints from the fatigue failure point of view and in this way contribute to increase the use of adhesive bonding over other joint techniques.
Resumo:
El magnesio y sus aleaciones representan un interesante campo de investigación dentro de la ingeniería de materiales debido a los retos que plantean tanto su conformabilidad como durabilidad. Las características físicas y mecánicas del magnesio y sus aleaciones los convierten en materiales de gran interés desde el punto de vista industrial al tratarse de uno de los materiales más abundantes y ligeros en un mundo en el que día a día los recursos y materias primas son más escasos por lo que el acceso a materiales abundantes y ligeros que permitan economizar el uso de energía cobrará mayor importancia en el futuro. En la coyuntura actual es por tanto necesario revisar procesos y procedimientos, investigando y tratando de ampliar desde este punto de vista si es posible mejorar los procedimientos de fabricación de los materiales disponibles actualmente o el desarrollo de nuevos, mejores que los anteriores, que permitan ayudar a la sostenibilidad del planeta. El magnesio, pese a ser un material muy abundante y relativamente barato, presenta una serie de inconvenientes que limitan de manera muy seria su aplicación industrial, su alta reactividad en presencia de oxígeno y su mal comportamiento frente a la corrosión así como limitaciones en su conformabilidad han estado limitando su uso y aplicaciones, los investigaciones dentro del campo de la metalurgia física de este material y el desarrollo de nuevas aleaciones han permitido su empleo en múltiples aplicaciones dentro de la industria aeroespacial, militar, automovilística, electrónica, deportiva y médica. La motivación para esta tesis doctoral ha sido tratar de aportar más luz sobre el comportamiento de una de las aleaciones comerciales base magnesio más empleadas, la AZ31B, tratando de modelizar como le afectan los procesos de soldadura y estudiando desde un punto de vista experimental como se ve modificada su microestructura, su comportamiento mecánico y su resistencia frente a la corrosión. Aunque en un principio se pensó en el empleo de métodos electroquímicos para el estudio de la corrosión de estos materiales, rápidamente se decidió prescindir de su uso dada la dificultad observada tanto durante los trabajos de investigación de esta Tesis como los encontrados por otros investigadores. Mediante microdurezas se han caracterizado mecánicamente las soldaduras de aleación de magnesio tipo AZ31 en función de diferentes materiales de aporte, observándose que el empleo de las aleaciones con mayor contenido de aluminio y zinc no contribuye a una mejora significativa de las propiedades mecánicas. Se han podido establecer correlaciones entre los modelos de simulación desarrollados y las microestructuras resultantes de los procesos reales de soldadura que permiten definir a priori que estructuras se van a obtener. De igual forma ha sido posible completar un estudio micrográfico y químico completo de las diferentes fases y microconstituyentes originados durante los procesos de soldadura, gracias a estos resultados se ha propuesto como hipótesis una explicación que justifica el comportamiento frente a la corrosión de estas aleaciones una vez soldadas. Los ensayos de corrosión realizados han permitido determinar correlaciones matemáticas que indican las velocidades de corrosión esperables de este tipo de aleaciones. Desde el punto de vista del diseño, los resultados obtenidos en este trabajo permitirán a otros investigadores y diseñadores tomar decisiones a la hora de decidir qué materiales de aporte emplear junto con las implicaciones que conllevan desde el punto de vista metalúrgico, mecánico o corrosivo las diferentes alternativas. Por último indicar que gracias al trabajo desarrollado se han definido modelos matemáticos para predecir el comportamiento frente a la corrosión de estas aleaciones, se han determinado las posibles causas y mecanismos por las que se gobierna la corrosión en la soldadura de chapas de aleación AZ31B y los motivos por los que se debe considerar el empleo de un material de aporte u otro. Los modelos de simulación desarrollados también han ayudado a comprender mejor la microestructura resultante de los procesos de soldadura y se han determinado que fases y microconstituyentes están presentes en las soldaduras de estas aleaciones. ABSTRACT Magnesium and its alloys represent and interesting research field in the material science due to the challenges of their fabrication and durability. The physical and mechanical properties of magnesium and its alloys make them a very interesting materials from and industrial point of view being one of the most abundant and lightest materials in a world in which day by day the lacking of resources and raw materials is more important, the use of light materials which allow to save energy will become more important in a near future. So that it is necessary to review processes and procedures, investigating and trying to improve current fabrication procedures and developing new ones, better than the former ones, in order to help with the sustainability of the planet. Although magnesium is a very common and relatively cheap material, it shows some inconveniences which limit in a major way their industrial application; its high reactivity in presence of oxygen, its poor corrosion resistance and some manufacturing problems had been limiting their use and applications, metallurgical investigations about this material and the development of new alloys have allowed its use in multiple applications in the aerospacial, military, automobile, electronics, sports and medical industry. The motivation for this thesis has been trying to clarify the behavior of one most used commercial base magnesium alloys, the AZ31, trying to modeling how its affected by thermal cycles of the welding process and studying from an experimental point of view how its microstructure is modified and how these modifications affect its mechanical behavior and corrosion resistance. Although at the beginning of this works it was though about the using of electrochemical techniques to evaluate the corrosion of these materials, rapidly it was decided not to use them because of the difficulty observed by during this research and by other investigators. The results obtained in this thesis have allowed to characterize mechanically AZ31 magnesium welding alloys considering different filler metals, according to this study using filler metals with a high content of aluminum and zinc does not represent an important improve It has been possible to establish correlations between simulation models and the resultant microstructures of the real melting processes originated during welding processes which allow to predict the structures which will be obtained after the welding. In addition to that it is possible to complete a complete micrographic and chemical analysis of the different phases and microconstituents created during welding, due to these results and hypothesis to explain the corrosion behavior of these welded alloys. Corrosion tests carried out have allowed defining mathematical correlations to predict corrosion rates of this kind of alloys. From a designing point of view, the results obtained in this work will let other investigators and designers to make decisions taking into account which implications have the different options from a metallurgical, mechanic and corrosive point of view. Finally we would like to indicate that thanks to this work it has been possible to define mathematical models to predict the corrosion behavior, the causes and the mechanism of this corrosion in the AZ31 welding sheets have been also determined and the reasons for using of one filler metal or another, the developed simulation models have also help to get a better understanding of the result microstructure determining the phases and the microconstituents present in the welding of this alloys.
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A proposal for an extended formulation of the power coefficient of a wind turbine is presented. This new formulation is a generalization of the Betz–Lanchester expression for the power coefficient as function of the axial deceleration of the wind speed provoked by the wind turbine in operation. The extended power coefficient takes into account the benefits of the power produced and the cost associated to the production of this energy. By the simple model proposed is evidenced that the purely energetic optimum operation condition giving rise to the Betz–Lanchester limit (maximum energy produced) does not coincide with the global optimum operational condition (maximum benefit generated) if cost of energy and degradation of the wind turbine during operation is considered. The new extended power coefficient is a general parameter useful to define global optimum operation conditions for wind turbines, considering not only the energy production but also the maintenance cost and the economic cost associated to the life reduction of the machine.
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La presente Tesis Doctoral evalúa la contribución de una fachada activa, constituida por acristalamientos con circulación de agua, en el rendimiento energético del edificio. Con especial énfasis en la baja afección sobre su imagen, su integración ha de favorecer la calificación del edificio con el futuro estándar de Edificio de consumo de Energía Casi Nulo (EECN). El propósito consiste en cuantificar su aportación a limitar la demanda de climatización, como solución de fachada transparente acorde a las normas de la energía del 2020. En el primer capítulo se introduce el planteamiento del problema. En el segundo capítulo se desarrollan la hipótesis y el objetivo fundamental de la investigación. Para tal fin, en el tercer capítulo, se revisa el estado del arte de la tecnología y de la investigación científica, mediante el análisis de la literatura de referencia. Se comparan patentes, prototipos, sistemas comerciales asimilables, investigaciones en curso en Universidades, y proyectos de investigación y desarrollo, sobre envolventes que incorporan acristalamientos con circulación de agua. El método experimental, expuesto en el cuarto capítulo, acomete el diseño, la fabricación y la monitorización de un prototipo expuesto, durante ciclos de ensayos, a las condiciones climáticas de Madrid. Esta fase ha permitido adquirir información precisa sobre el rendimiento del acristalamiento en cada orientación de incidencia solar, en las distintas estaciones del año. En paralelo, se aborda el desarrollo de modelos teóricos que, mediante su asimilación a soluciones multicapa caracterizadas en las herramientas de simulación EnergyPlus y IDA-ICE (IDA Indoor Climate and Energy), reproducen el efecto experimental. En el quinto capítulo se discuten los resultados experimentales y teóricos, y se analiza la respuesta del acristalamiento asociado a un determinado volumen y temperatura del agua. Se calcula la eficiencia en la captación de la radiación y, mediante la comparativa con un acristalamiento convencional, se determina la reducción de las ganancias solares y las pérdidas de energía. Se comparan el rendimiento del acristalamiento, obtenido experimentalmente, con el ofrecido por paneles solares fototérmicos disponibles en el mercado. Mediante la traslación de los resultados experimentales a casos de células de tamaño habitable, se cuantifica la afección del acristalamiento sobre el consumo en refrigeración y calefacción. Diferenciando cada caso por su composición constructiva y orientación, se extraen conclusiones sobre la reducción del gasto en climatización, en condiciones de bienestar. Posteriormente, se evalúa el ahorro de su incorporación en un recinto existente, de construcción ligera, localizado en la Escuela de Arquitectura de la Universidad Politécnica de Madrid (UPM). Mediante el planteamiento de escenarios de rehabilitación energética, se estima su compatibilidad con un sistema de climatización mediante bomba de calor y extracción geotérmica. Se describe el funcionamiento del sistema, desde la perspectiva de la operación conjunta de los acristalamientos activos e intercambio geotérmico, en nuestro clima. Mediante la parametrización de sus funciones, se estima el beneficio adicional de su integración, a partir de la mejora del rendimiento de la bomba de calor COP (Coefficient of Performance) en calefacción, y de la eficiencia EER (Energy Efficiency Ratio) en refrigeración. En el recinto de la ETSAM, se ha analizado la contribución de la fachada activa en su calificación como Edificio de Energía Casi Nula, y estudiado la rentabilidad económica del sistema. En el sexto capítulo se exponen las conclusiones de la investigación. A la fecha, el sistema supone alta inversión inicial, no obstante, genera elevada eficiencia con bajo impacto arquitectónico, reduciéndose los costes operativos, y el dimensionado de los sistemas de producción, de mayor afección sobre el edificio. Mediante la envolvente activa con suministro geotérmico no se condena la superficie de cubierta, no se ocupa volumen útil por la presencia de equipos emisores, y no se reduce la superficie o altura útil a base de reforzar los aislamientos. Tras su discusión, se considera una alternativa de valor en procesos de diseño y construcción de Edificios de Energía Casi Nulo. Se proponen líneas de futuras investigación cuyo propósito sea el conocimiento de la tecnología de los acristalamientos activos. En el último capítulo se presentan las actividades de difusión de la investigación. Adicionalmente se ha proporcionado una mejora tecnológica a las fachadas activas existentes, que ha derivado en la solicitud de una patente, actualmente en tramitación. ABSTRACT This Thesis evaluates the contribution of an active water flow glazing façade on the energy performance of buildings. Special emphasis is made on the low visual impact on its image, and the active glazing implementation has to encourage the qualification of the building with the future standard of Nearly Zero Energy Building (nZEB). The purpose is to quantify the façade system contribution to limit air conditioning demand, resulting in a transparent façade solution according to the 2020 energy legislation. An initial approach to the problem is presented in first chapter. The second chapter develops the hypothesis and the main objective of the research. To achieve this purpose, the third chapter reviews the state of the art of the technology and scientific research, through the analysis of reference literature. Patents, prototypes, assimilable commercial systems, ongoing research in other universities, and finally research and development projects incorporating active fluid flow glazing are compared. The experimental method, presented in fourth chapter, undertakes the design, manufacture and monitoring of a water flow glazing prototype exposed during test cycles to weather conditions in Madrid. This phase allowed the acquisition of accurate information on the performance of water flow glazing on each orientation of solar incidence, during different seasons. In parallel, the development of theoretical models is addressed which, through the assimilation to multilayer solutions characterized in the simulation tools EnergyPlus and IDA-Indoor Climate and Energy, reproduce the experimental effect. Fifth chapter discusses experimental and theoretical results focused to the analysis of the active glazing behavior, associated with a specific volume and water flow temperature. The efficiency on harvesting incident solar radiation is calculated, and, by comparison with a conventional glazing, the reduction of solar gains and energy losses are determined. The experimental performance of fluid flow glazing against the one offered by photothermal solar panels available on the market are compared. By translating the experimental and theoretical results to cases of full-size cells, the reduction in cooling and heating consumption achieved by active fluid glazing is quantified. The reduction of energy costs to achieve comfort conditions is calculated, differentiating each case by its whole construction composition and orientation. Subsequently, the saving of the implementation of the system on an existing lightweight construction enclosure, located in the School of Architecture at the Polytechnic University of Madrid (UPM), is then calculated. The compatibility between the active fluid flow glazing and a heat pump with geothermal heat supply system is estimated through the approach of different energy renovation scenarios. The overall system operation is described, from the perspective of active glazing and geothermal heat exchange combined operation, in our climate. By parameterization of its functions, the added benefit of its integration it is discussed, particularly from the improvement of the heat pump performance COP (Coefficient of Performance) in heating and efficiency EER (Energy Efficiency Ratio) in cooling. In the case study of the enclosure in the School of Architecture, the contribution of the active glazing façade in qualifying the enclosure as nearly Zero Energy Building has been analyzed, and the feasibility and profitability of the system are studied. The sixth chapter sets the conclusions of the investigation. To date, the system may require high initial investment; however, high efficiency with low architectural impact is generated. Operational costs are highly reduced as well as the size and complexity of the energy production systems, which normally have huge visual impact on buildings. By the active façade with geothermal supply, the deck area it is not condemned. Useful volume is not consumed by the presence of air-conditioning equipment. Useful surface and room height are not reduced by insulation reinforcement. After discussion, water flow glazing is considered a potential value alternative in nZEB design and construction processes. Finally, this chapter proposes future research lines aiming to increase the knowledge of active water flow glazing technology. The last chapter presents research dissemination activities. Additionally, a technological improvement to existing active facades has been developed, which has resulted in a patent application, currently in handling process.
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Restenosis continues to be a major problem limiting the effectiveness of revascularization procedures. To date, the roles of heterotrimeric G proteins in the triggering of pathological vascular smooth muscle (VSM) cell proliferation have not been elucidated. βγ subunits of heterotrimeric G proteins (Gβγ) are known to activate mitogen-activated protein (MAP) kinases after stimulation of certain G protein-coupled receptors; however, their relevance in VSM mitogenesis in vitro or in vivo is not known. Using adenoviral-mediated transfer of a transgene encoding a peptide inhibitor of Gβγ signaling (βARKct), we evaluated the role of Gβγ in MAP kinase activation and proliferation in response to several mitogens, including serum, in cultured rat VSM cells. Our results include the striking finding that serum-induced proliferation of VSM cells in vitro is mediated largely via Gβγ. Furthermore, we studied the effects of in vivo adenoviral-mediated βARKct gene transfer on VSM intimal hyperplasia in a rat carotid artery restenosis model. Our in vivo results demonstrated that the presence of the βARKct in injured rat carotid arteries significantly reduced VSM intimal hyperplasia by 70%. Thus, Gβγ plays a critical role in physiological VSM proliferation, and targeted Gβγ inhibition represents a novel approach for the treatment of pathological conditions such as restenosis.
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The last few years have witnessed a significant decrease in the gap between the Shannon channel capacity limit and what is practically achievable. Progress has resulted from novel extensions of previously known coding techniques involving interleaved concatenated codes. A considerable body of simulation results is now available, supported by an important but limited theoretical basis. This paper presents a computational technique which further ties simulation results to the known theory and reveals a considerable reduction in the complexity required to approach the Shannon limit.
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A novel type I transmembrane protein of COPI-coated vesicles, p23, has been demonstrated to be localized mainly to the Golgi complex. This protein and p24, another member of the p24 family, have been shown to bind coatomer via their short cytoplasmic tails. Here we demonstrate that p23 continuously cycles through the early secretory pathway. The cytoplasmic tail of p23 is shown to act as a functional retrieval signal as it confers endoplasmic reticulum (ER) residence to a CD8–p23 fusion protein. This ER localization is, at least in part, a result of retrieval from post-ER compartments because CD8–p23 fusion proteins receive post-ER modifications. In contrast, the cytoplasmic tail of p24 has been shown not to retrieve a CD8–p24 fusion protein. The coatomer binding motifs FF and KK in the cytoplasmic tail of p23 are reported to influence the steady-state localization of the CD8–p23 fusion protein within the ER–Golgi recycling pathway. It appears that the steady-state Golgi localization of endogenous p23 is maintained by its lumenal domain, as a fusion protein with the lumenal domain of CD8, and the membrane span as well as the cytoplasmic tail of p23 is no longer detected in the Golgi.
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Formulas are derived for the effect of size on a free-swimming microbe’s ability to follow chemical, light, or temperature stimuli or to disperse in random directions. The four main assumptions are as follows: (i) the organisms can be modeled as spheres, (ii) the power available to the organism for swimming is proportional to its volume, (iii) the noise in measuring a signal limits determination of the direction of a stimulus, and (iv) the time available to determine stimulus direction or to swim a straight path is limited by rotational diffusion caused by Brownian motion. In all cases, it is found that there is a sharp size limit below which locomotion has no apparent benefit. This size limit is estimated to most probably be about 0.6 μm diameter and is relatively insensitive to assumed values of the other parameters. A review of existing descriptions of free-floating bacteria reveals that the smallest of 97 motile genera has a mean length of 0.8 μm, whereas 18 of 94 nonmotile genera are smaller. Similar calculations have led to the conclusion that a minimum size also exists for use of pheromones in mate location, although this size limit is about three orders of magnitude larger. In both cases, the application of well-established physical laws and biological generalities has demonstrated that a common feature of animal behavior is of no use to small free-swimming organisms.
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Kinesin is a processive motor protein: A single molecule can walk continuously along a microtubule for several micrometers, taking hundreds of 8-nm steps without dissociating. To elucidate the biochemical and structural basis for processivity, we have engineered a heterodimeric one-headed kinesin and compared its biochemical properties to those of the wild-type two-headed molecule. Our construct retains the functionally important neck and tail domains and supports motility in high-density microtubule gliding assays, though it fails to move at the single-molecule level. We find that the ATPase rate of one-headed kinesin is 3–6 s−1 and that detachment from the microtubule occurs at a similar rate (3 s−1). This establishes that one-headed kinesin usually detaches once per ATP hydrolysis cycle. Furthermore, we identify the rate-limiting step in the one-headed hydrolysis cycle as detachment from the microtubule in the ADP⋅Pi state. Because the ATPase and detachment rates are roughly an order of magnitude lower than the corresponding rates for two-headed kinesin, the detachment of one head in the homodimer (in the ADP⋅Pi state) must be accelerated by the other head. We hypothesize that this results from internal strain generated when the second head binds. This idea accords with a hand-over-hand model for processivity in which the release of the trailing head is contingent on the binding of the forward head. These new results, together with previously published ones, allow us to propose a pathway that defines the chemical and mechanical cycle for two-headed kinesin.
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The MAL proteolipid is a nonglycosylated integral membrane protein found in glycolipid-enriched membrane microdomains. In polarized epithelial Madin-Darby canine kidney cells, MAL is necessary for normal apical transport and accurate sorting of the influenza virus hemagglutinin. MAL is thus part of the integral machinery for glycolipid-enriched membrane–mediated apical transport. At steady state, MAL is predominantly located in perinuclear vesicles that probably arise from the trans-Golgi network (TGN). To act on membrane traffic and to prevent their accumulation in the target compartment, integral membrane elements of the protein-sorting machinery should be itinerant proteins that cycle between the donor and target compartments. To establish whether MAL is an itinerant protein, we engineered the last extracellular loop of MAL by insertion of sequences containing the FLAG epitope or with sequences containing residues that became O-glycosylated within the cells or that displayed biotinylatable groups. The ectopic expression of these modified MAL proteins allowed us to investigate the surface expression of MAL and its movement through different compartments after internalization with the use of a combination of assays, including surface biotinylation, surface binding of anti-FLAG antibodies, neuraminidase sensitivity, and drug treatments. Immunofluorescence and flow cytometric analyses indicated that, in addition to its Golgi localization, MAL was also expressed on the cell surface, from which it was rapidly internalized. This retrieval implies transport through the endosomal pathway and requires endosomal acidification, because it can be inhibited by drugs such as chloroquine, monensin, and NH4Cl. Resialylation experiments of surface MAL treated with neuraminidase indicated that ∼30% of the internalized MAL molecules were delivered to the TGN, probably to start a new cycle of cargo transport. Together, these observations suggest that, as predicted for integral membrane members of the late protein transport machinery, MAL is an itinerant protein cycling between the TGN and the plasma membrane.
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The hair follicle cycle successively goes through the anagen, catagen, telogen, and latency phases, which correspond, respectively, to hair growth, arrest, shedding, and absence before a new anagen phase is initiated. Experimental observations collected over a period of 14 years in a group of 10 male volunteers, alopecic and nonalopecic, allowed us to determine the characteristics of scalp hair follicle cycles. On the basis of these observations, we propose a follicular automaton model to simulate the dynamics of human hair cycles. The automaton model is defined by a set of rules that govern the stochastic transitions of each follicle between the successive states anagen, telogen, and latency, and the subsequent return to anagen. The transitions occur independently for each follicle, after time intervals given stochastically by a distribution characterized by a mean and a variance. The follicular automaton model accounts both for the dynamical transitions observed in a single follicle and for the behavior of an ensemble of independently cycling follicles. Thus, the model successfully reproduces the evolution of the fractions of follicle populations in each of the three phases, which fluctuate around steady-state or slowly drifting values. We apply the follicular automaton model to the study of spatial patterns of follicular growth that result from a spatially heterogeneous distribution of parameters such as the mean duration of anagen phase. When considering that follicles die or miniaturize after going through a critical number of successive cycles, the model can reproduce the evolution to hair patterns similar to well known types of diffuse or androgenetic alopecia.
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The single gene encoding limit dextrinase (pullulan 6-glucanohydrolase; EC 3.2.1.41) in barley (Hordeum vulgare) has 26 introns that range in size from 93 to 822 base pairs. The mature polypeptide encoded by the gene has 884 amino acid residues and a calculated molecular mass of 97,417 D. Limit dextrinase mRNA is abundant in gibberellic acid-treated aleurone layers and in germinated grain. Gibberellic acid response elements were found in the promoter region of the gene. These observations suggest that the enzyme participates in starch hydrolysis during endosperm mobilization in germinated grain. The mRNA encoding the enzyme is present at lower levels in the developing endosperm of immature grain, a location consistent with a role for limit dextrinase in starch synthesis. Enzyme activity was also detected in developing grain. The limit dextrinase has a presequence typical of transit peptides that target nascent polypeptides to amyloplasts, but this would not be expected to direct secretion of the mature enzyme from aleurone cells in germinated grain. It remains to be discovered how the enzyme is released from the aleurone and whether another enzyme, possibly of the isoamylase group, might be equally important for starch hydrolysis in germinated grain.
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Neuronal progenitors and tumor cells possess propensity to proliferate and to migrate. Glutamate regulates proliferation and migration of neurons during development, but it is not known whether it influences proliferation and migration of tumor cells. We demonstrate that glutamate antagonists inhibit proliferation of human tumor cells. Colon adenocarcinoma, astrocytoma, and breast and lung carcinoma cells were most sensitive to the antiproliferative effect of the N-methyl-d-aspartate antagonist dizocilpine, whereas breast and lung carcinoma, colon adenocarcinoma, and neuroblastoma cells responded most favorably to the α-amino-3-hydroxy-5-methyl-4-isoxazole-propionate antagonist GYKI52466. The antiproliferative effect of glutamate antagonists was Ca2+ dependent and resulted from decreased cell division and increased cell death. Morphological alterations induced by glutamate antagonists in tumor cells consisted of reduced membrane ruffling and pseudopodial protrusions. Furthermore, glutamate antagonists decreased motility and invasive growth of tumor cells. These findings suggest anticancer potential of glutamate antagonists.