973 resultados para Structural dynamics.


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Bridges with deck supported on either sliding or elastomeric bearings are very common in mid-seismicity regions. Their main seismic vulnerabilities are related to the pounding of the deck against abutments or between the different deck elements. A simplified model of the longitudinal behavior of those bridges will allow to characterize the reaction forces developed during pounding using the Pacific Earthquake Engineering Research Center framework formula. In order to ensure the general applicability of the results obtained, a large number of system parameter combinations will be considered. The heart of the formula is the identification of suitable intermediate variables. First, the pseudo acceleration spectral value for the fundamental period of the system (Sa(Ts)) will be used as an intensity measure (IM). This IM will result in a very large non-explained variability of the engineering demand parameter. A portion of this variability will be proved to be related to the relative content of high-frequency energy in the input motion. Two vector-valued IMs including a second parameter taking this energy content into account will then be considered. For both of them, a suitable form for the conditional intensity dependence of the response will be obtained. The question of which one to choose will also be analyzed. Finally, additional issues related to the IM will be studied: its applicability to pulse-type records, the validity of scaling records and the sufficiency of the IM.

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This paper presents the results of part of the research carried out by a committee in charge of the elaboration of the new Spanish Code of Actions in Railway Bridges. Following the work developed by the European Rail Research Institute (ERRI), the dynamic effects caused by the Spanish high-speed train TALGO have been studied and compared with other European trains. A simplified envelope of the impact coefficient is also presented. Finally, the train-bridge interactions has been analysed and the results compared with those obtained from simple models based on moving loads.

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This paper presents a time-domain stochastic system identification method based on Maximum Likelihood Estimation and the Expectation Maximization algorithm. The effectiveness of this structural identification method is evaluated through numerical simulation in the context of the ASCE benchmark problem on structural health monitoring. Modal parameters (eigenfrequencies, damping ratios and mode shapes) of the benchmark structure have been estimated applying the proposed identification method to a set of 100 simulated cases. The numerical results show that the proposed method estimates all the modal parameters reasonably well in the presence of 30% measurement noise even. Finally, advantages and disadvantages of the method have been discussed.

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In Operational Modal Analysis (OMA) of a structure, the data acquisition process may be repeated many times. In these cases, the analyst has several similar records for the modal analysis of the structure that have been obtained at di�erent time instants (multiple records). The solution obtained varies from one record to another, sometimes considerably. The differences are due to several reasons: statistical errors of estimation, changes in the external forces (unmeasured forces) that modify the output spectra, appearance of spurious modes, etc. Combining the results of the di�erent individual analysis is not straightforward. To solve the problem, we propose to make the joint estimation of the parameters using all the records. This can be done in a very simple way using state space models and computing the estimates by maximum-likelihood. The method provides a single result for the modal parameters that combines optimally all the records.

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The design of containment walls suffering seismic loads traditionally has been realized with methods based on pseudoanalitic procedures such as Mononobe-Okabe's method, which it has led in certain occasions to insecure designs, that they have produced the ruin of many containment walls suffering the action of an earthquake. The recommendations gathered in Mononobe-Okabe's theory have been included in numerous Codes of Seismic Design. It is clear that a revision of these recommendations must be done. At present there is taking place an important review of the design methods of anti-seismic structures such as containment walls placed in an area of numerous earthquakes, by means of the introduction at the beginning of the decade of 1990 the Displacement Response Spectrum (DRS) and the Capacity Demand Diagram (CDD) that suppose an important change in the way of presenting the Elastic Response Spectrum (ERS). On the other hand in case of action of an earthquake, the dynamic characteristics of a soil have been referred traditionally to the speed of the shear waves that can be generated in a site, together with the characteristics of plasticity and damping of the soil. The Principle of the energy conservation explains why a shear upward propagating seismic wave can be amplified when travelling from a medium with high shear wave velocity (rock) to other medium with lower velocity (soil deposit), as it happened in the earthquake of Mexico of 1985. This amplification is a function of the speed gradient or of the contrast of impedances in the border of both types of mediums. A method is proposed in this paper for the design of containment walls in different soils, suffering to the action of an earthquake, based on the Performance-Based Seismic Design.

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This paper presents shake-table tests conducted on a two-fifths-scale reinforced concrete frame representing a conventional construction design under current building code provisions in the Mediterranean area. The structure was subjected to a sequence of dynamic tests including free vibrations and four seismic simulations in which a historical ground motion record was scaled to levels of increasing intensity until collapse. Each seismic simulation was associated with a different level of seismic hazard, representing very frequent, frequent, rare and very rare earthquakes. The structure remained basically undamaged and within the inter-story drift limits of the "immediate occupancy" performance level for the very frequent and frequent earthquakes. For the rare earthquake, the specimen sustained significant damage with chord rotations of up to 28% of its ultimate capacity and approached the upper bound limit of inter-story drift associated with "life safety". The specimen collapsed at the beginning of the "very rare" seismic simulation. Besides summarizing the experimental program, this paper evaluates the damage quantitatively at the global and local levels in terms of chord rotation and other damage indexes, together with the energy dissipation demands for each level of seismic hazard. Further, the ratios of column-to-beam moment capacity recommended by Eurocode 8 and ACI-318 to guarantee the formation of a strong column-weak beam mechanism are examined.

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Passive energy dissipation devices are increasingly implemented in frame structures to improve their performance under seismic loading. Most guidelines for designing this type of system retain the requirements applicable to frames without dampers, and this hinders taking full advantage of the benefits of implementing dampers. Further, assessing the extent of damage suffered by the frame and by the dampers for different levels of seismic hazard is of paramount importance in the framework of performance-based design. This paper presents an experimental investigation whose objectives are to provide empirical data on the response of reinforced concrete (RC) frames equipped with hysteretic dampers (dynamic response and damage) and to evaluate the need for the frame to form a strong column-weak beam mechanism and dissipate large amounts of plastic strain energy. To this end, shake-table tests were conducted on a 2/5-scale RC frame with hysteretic dampers. The frame was designed only for gravitational loads. The dampers provided lateral strength and stiffness, respectively, three and 12 times greater than those of the frame. The test structure was subjected to a sequence of seismic simulations that represented different levels of seismic hazard. The RC frame showed a performance level of "immediate occupancy", with maximum rotation demands below 20% of the ultimate capacity. The dampers dissipated most of the energy input by the earthquake. It is shown that combining hysteretic dampers with flexible reinforced concrete frames leads to structures with improved seismic performance and that requirements of conventional RC frames (without dampers) can be relieved.

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The paper reports on a collaborative effort between the Swiss Federal Nuclear Safety Inspectorate (ENSI) and their consultants Principia and Stangenberg. As part of the IMPACT III project, reduced scale impact tests of reinforced concrete structures were carried out. The simulation of test X3 is presented here and the numerical results are compared with those obtained in the test, carried out in August 2013. The general object is to improve the safety of nuclear facilities and, more specifically, to demonstrate the capabilities of current simulation techniques to reproduce the behaviour of a reinforced concrete structure impacted by a soft missile. The missile is a steel tube with a mass of 50 kg and travelling at 140 m/s. The target is a 250 mm thick, 2,1 m by 2,1 m reinforced concrete wall, held in a stiff supporting frame. The reinforcement includes both longitudinal and transverse rebars. Calculations were carried out before and after the test with Abaqus (Principia) and SOFiSTiK (Stangenberg). In the Abaqus simulation the concrete is modelled using solid elements and a damaged plasticity formulation, the rebars with embedded beam elements, and the missile with shell elements. In SOFiSTiK the target is modelled with non-linear, layered shell elements for the reinforcement on both sides; non-linear shear deformations of shell/plate elements are approximately included. The results generally indicate a good agreement between calculations and measurements.

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The application of the Electro-Mechanical Impedance (EMI) method for damage detection in Structural Health Monitoring has noticeable increased in recent years. EMI method utilizes piezoelectric transducers for directly measuring the mechanical properties of the host structure, obtaining the so called impedance measurement, highly influenced by the variations of dynamic parameters of the structure. These measurements usually contain a large number of frequency points, as well as a high number of dimensions, since each frequency range swept can be considered as an independent variable. That makes this kind of data hard to handle, increasing the computational costs and being substantially time-consuming. In that sense, the Principal Component Analysis (PCA)-based data compression has been employed in this work, in order to enhance the analysis capability of the raw data. Furthermore, a Support Vector Machine (SVM), which has been widespread used in machine learning and pattern recognition fields, has been applied in this study in order to model any possible existing pattern in the PCAcompress data, using for that just the first two Principal Components. Different known non-damaged and damaged measurements of an experimental tested beam were used as training input data for the SVM algorithm, using as test input data the same amount of cases measured in beams with unknown structural health conditions. Thus, the purpose of this work is to demonstrate how, with a few impedance measurements of a beam as raw data, its healthy status can be determined based on pattern recognition procedures.

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Arch bridge structural solution has been known for centuries, in fact the simple nature of arch that require low tension and shear strength was an advantage as the simple materials like stone and brick were the only option back in ancient centuries. By the pass of time especially after industrial revolution, the new materials were adopted in construction of arch bridges to reach longer spans. Nowadays one long span arch bridge is made of steel, concrete or combination of these two as "CFST", as the result of using these high strength materials, very long spans can be achieved. The current record for longest arch belongs to Chaotianmen bridge over Yangtze river in China with 552 meters span made of steel and the longest reinforced concrete type is Wanxian bridge which also cross the Yangtze river through a 420 meters span. Today the designer is no longer limited by span length as long as arch bridge is the most applicable solution among other approaches, i.e. cable stayed and suspended bridges are more reasonable if very long span is desired. Like any super structure, the economical and architectural aspects in construction of a bridge is extremely important, in other words, as a narrower bridge has better appearance, it also require smaller volume of material which make the design more economical. Design of such bridge, beside the high strength materials, requires precise structural analysis approaches capable of integrating the combination of material behaviour and complex geometry of structure and various types of loads which may be applied to bridge during its service life. Depend on the design strategy, analysis may only evaluates the linear elastic behaviour of structure or consider the nonlinear properties as well. Although most of structures in the past were designed to act in their elastic range, the rapid increase in computational capacity allow us to consider different sources of nonlinearities in order to achieve a more realistic evaluations where the dynamic behaviour of bridge is important especially in seismic zones where large movements may occur or structure experience P - _ effect during the earthquake. The above mentioned type of analysis is computationally expensive and very time consuming. In recent years, several methods were proposed in order to resolve this problem. Discussion of recent developments on these methods and their application on long span concrete arch bridges is the main goal of this research. Accordingly available long span concrete arch bridges have been studied to gather the critical information about their geometrical aspects and properties of their materials. Based on concluded information, several concrete arch bridges were designed for further studies. The main span of these bridges range from 100 to 400 meters. The Structural analysis methods implemented in in this study are as following: Elastic Analysis: Direct Response History Analysis (DRHA): This method solves the direct equation of motion over time history of applied acceleration or imposed load in linear elastic range. Modal Response History Analysis (MRHA): Similar to DRHA, this method is also based on time history, but the equation of motion is simplified to single degree of freedom system and calculates the response of each mode independently. Performing this analysis require less time than DRHA. Modal Response Spectrum Analysis (MRSA): As it is obvious from its name, this method calculates the peak response of structure for each mode and combine them using modal combination rules based on the introduced spectra of ground motion. This method is expected to be fastest among Elastic analysis. Inelastic Analysis: Nonlinear Response History Analysis (NL-RHA): The most accurate strategy to address significant nonlinearities in structural dynamics is undoubtedly the nonlinear response history analysis which is similar to DRHA but extended to inelastic range by updating the stiffness matrix for every iteration. This onerous task, clearly increase the computational cost especially for unsymmetrical buildings that requires to be analyzed in a full 3D model for taking the torsional effects in to consideration. Modal Pushover Analysis (MPA): The Modal Pushover Analysis is basically the MRHA but extended to inelastic stage. After all, the MRHA cannot solve the system of dynamics because the resisting force fs(u; u_ ) is unknown for inelastic stage. The solution of MPA for this obstacle is using the previously recorded fs to evaluate system of dynamics. Extended Modal Pushover Analysis (EMPA): Expanded Modal pushover is a one of very recent proposed methods which evaluates response of structure under multi-directional excitation using the modal pushover analysis strategy. In one specific mode,the original pushover neglect the contribution of the directions different than characteristic one, this is reasonable in regular symmetric building but a structure with complex shape like long span arch bridges may go through strong modal coupling. This method intend to consider modal coupling while it take same time of computation as MPA. Coupled Nonlinear Static Pushover Analysis (CNSP): The EMPA includes the contribution of non-characteristic direction to the formal MPA procedure. However the static pushovers in EMPA are performed individually for every mode, accordingly the resulted values from different modes can be combined but this is only valid in elastic phase; as soon as any element in structure starts yielding the neutral axis of that section is no longer fixed for both response during the earthquake, meaning the longitudinal deflection unavoidably affect the transverse one or vice versa. To overcome this drawback, the CNSP suggests executing pushover analysis for governing modes of each direction at the same time. This strategy is estimated to be more accurate than MPA and EMPA, moreover the calculation time is reduced because only one pushover analysis is required. Regardless of the strategy, the accuracy of structural analysis is highly dependent on modelling and numerical integration approaches used in evaluation of each method. Therefore the widely used Finite Element Method is implemented in process of all analysis performed in this research. In order to address the study, chapter 2, starts with gathered information about constructed long span arch bridges, this chapter continuous with geometrical and material definition of new models. Chapter 3 provides the detailed information about structural analysis strategies; furthermore the step by step description of procedure of all methods is available in Appendix A. The document ends with the description of results and conclusion of chapter 4.

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Operational Modal Analysis consists on estimate the modal parameters of a structure (natural frequencies, damping ratios and modal vectors) from output-only vibration measurements. The modal vectors can be only estimated where a sensor is placed, so when the number of available sensors is lower than the number of tested points, it is usual to perform several tests changing the position of the sensors from one test to the following (multiple setups of sensors): some sensors stay at the same position from setup to setup, and the other sensors change the position until all the tested points are covered. The permanent sensors are then used to merge the mode shape estimated at each setup (or partial modal vectors) into global modal vectors. Traditionally, the partial modal vectors are estimated independently setup by setup, and the global modal vectors are obtained in a postprocess phase. In this work we present two state space models that can be used to process all the recorded setups at the same time, and we also present how these models can be estimated using the maximum likelihood method. The result is that the global mode shape of each mode is obtained automatically, and subsequently, a single value for the natural frequency and damping ratio of the mode is computed. Finally, both models are compared using real measured data.

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La dinámica estructural estudia la respuesta de una estructura ante cargas o fenómenos variables en el tiempo. En muchos casos, estos fenómenos requieren realizar análisis paramétricos de la estructura considerando una gran cantidad de configuraciones de diseño o modificaciones de la estructura. Estos cambios, ya sean en fases iniciales de diseño o en fases posteriores de rediseño, alteran las propiedades físicas de la estructura y por tanto del modelo empleado para su análisis, cuyo comportamiento dinámico se modifica en consecuencia. Un caso de estudio de este tipo de modificaciones es la supervisión de la integridad estructural, que trata de identificar la presencia de daño estructural y prever el comportamiento de la estructura tras ese daño, como puede ser la variación del comportamiento dinámico de la estructura debida a una delaminación, la aparición o crecimiento de grieta, la debida a la pérdida de pala sufrida por el motor de un avión en vuelo, o la respuesta dinámica de construcciones civiles como puentes o edificios frente a cargas sísmicas. Si a la complejidad de los análisis dinámicos requeridos en el caso de grandes estructuras se añade la variación de determinados parámetros en busca de una respuesta dinámica determinada o para simular la presencia de daños, resulta necesario la búsqueda de medios de simplificación o aceleración del conjunto de análisis que de otra forma parecen inabordables tanto desde el punto de vista del tiempo de computación, como de la capacidad requerida de almacenamiento y manejo de grandes volúmenes de archivos de datos. En la presente tesis doctoral se han revisado los métodos de reducción de elementos .nitos más habituales para análisis dinámicos de grandes estructuras. Se han comparado los resultados de casos de estudio de los métodos más aptos, para el tipo de estructuras y modificaciones descritas, con los resultados de aplicación de un método de reducción reciente. Entre los primeros están el método de condensación estática de Guyan extendido al caso con amortiguamiento no proporcional y posteriores implementaciones de condensaciones dinámicas en diferentes espacios vectoriales. El método de reducción recientemente presentado se denomina en esta tesis DACMAM (Dynamic Analysis in Complex Modal space Acceleration Method), y consiste en el análisis simplificado que proporciona una solución para la respuesta dinámica de una estructura, calculada en el espacio modal complejo y que admite modificaciones estructurales. El método DACMAM permite seleccionar un número reducido de grados de libertad significativos para la dinámica del fenómeno que se quiere estudiar como son los puntos de aplicación de la carga, localizaciones de los cambios estructurales o puntos donde se quiera conocer la respuesta, de forma que al implementar las modificaciones estructurales, se ejecutan los análisis necesarios sólo de dichos grados de libertad sin pérdida de precisión. El método permite considerar alteraciones de masa, rigidez, amortiguamiento y la adición de nuevos grados de libertad. Teniendo en cuenta la dimensión del conjunto de ecuaciones a resolver, la parametrización de los análisis no sólo resulta posible, sino que es también manejable y controlable gracias a la sencilla implementación del procedimiento para los códigos habituales de cálculo mediante elementos .nitos. En el presente trabajo se muestra la bondad y eficiencia del método en comparación con algunos de los métodos de reducción de grandes modelos estructurales, verificando las diferencias entre sí de los resultados obtenidos y respecto a la respuesta real de la estructura, y comprobando los medios empleados en ellos tanto en tiempo de ejecución como en tamaño de ficheros electrónicos. La influencia de los diversos factores que se tienen en cuenta permite identificar los límites y capacidades de aplicación del método y su exhaustiva comparación con los otros procedimientos. ABSTRACT Structural dynamics studies the response of a structure under loads or phenomena which vary over time. In many cases, these phenomena require the use of parametric analyses taking into consideration several design configurations or modifications of the structure. This is a typical need in an engineering o¢ ce, no matter the structural design is in early or final stages. These changes modify the physical properties of the structure, and therefore, the finite element model to analyse it. A case study, that exempli.es this circumstance, is the structural health monitoring to predict the variation of the dynamical behaviour after damage, such as a delaminated structure, a crack onset or growth, an aircraft that suffers a blade loss event or civil structures (buildings or bridges) under seismic loads. Not only large structures require complex analyses to appropriately acquire an accurate solution, but also the variation of certain parameters. There is a need to simplify the analytical process, in order to bring CPU time, data .les, management of solutions to a reasonable size. In the current doctoral thesis, the most common finite element reduction methods for large structures are reviewed. Results of case studies are compared between a recently proposed method, herein named DACMAM (Dynamic Analysis in Complex Modal space Acceleration Method), and different condensation methods, namely static or Guyan condensation and dynamic condensation in different vectorial spaces. All these methods are suitable for considering non-classical damping. The reduction method DACMAM consist of a structural modification in the complex modal domain which provides a dynamic response solution for the reduced models. This process allows the selection of a few degrees of freedom that are relevant for the dynamic response of the system. These d.o.f. are the load application points, relevant structural points or points in which it is important to know the response. Consequently, an analysis with structural modifications implies only the calculation of the dynamic response of the selected degrees of freedom added, but with no loss of information. Therefore, mass, stiffness or damping modifications are easily considered as well as new degrees of freedom. Taking into account the size of the equations to be solved, the parameterization of the dynamic solutions is not only possible, but also manageable and controllable due to the easy implementation of the procedure in the standard finite element solvers. In this thesis, the proposed reduction method for large structural models is compared with other published model order reduction methods. The comparison shows and underlines the efficiency of the new method, and veri.es the differences in the response when compared with the response of the full model. The CPU time, the data files and the scope of the parameterization are also addressed.

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Los fenómenos dinámicos pueden poner en peligro la integridad de estructuras aeroespaciales y los ingenieros han desarrollado diferentes estrategias para analizarlos. Uno de los grandes problemas que se plantean en la ingeniería es cómo atacar un problema dinámico estructural. En la presente tesis se plantean distintos fenómenos dinámicos y se proponen métodos para estimar o simular sus comportamientos mediante un análisis paramétrico determinista y aleatorio del problema. Se han propuesto desde problemas sencillos con pocos grados de libertad que sirven para analizar las diferentes estrategias y herramientas a utilizar, hasta fenómenos muy dinámicos que contienen comportamientos no lineales, daños y fallos. Los primeros ejemplos de investigación planteados cubren una amplia gama de los fenómenos dinámicos, como el análisis de vibraciones de elementos másicos, incluyendo impactos y contactos, y el análisis de una viga con carga armónica aplicada a la que también se le añaden parámetros aleatorios que pueden responder a un desconocimiento o incertidumbre de los mismos. Durante el desarrollo de la tesis se introducen conceptos y se aplican distintos métodos, como el método de elementos finitos (FEM) en el que se analiza su resolución tanto por esquemas implícitos como explícitos, y métodos de análisis paramétricos y estadísticos mediante la técnica de Monte Carlo. Más adelante, una vez ya planteadas las herramientas y estrategias de análisis, se estudian fenómenos más complejos, como el impacto a baja velocidad en materiales compuestos, en el que se busca evaluar la resistencia residual y, por lo tanto, la tolerancia al daño de la estructura. Se trata de un suceso que puede producirse por la caída de herramienta, granizo o restos en la pista de aterrizaje. Otro de los fenómenos analizados también se da en un aeropuerto y se trata de la colisión con un dispositivo frangible, el cual tiene que romperse bajo ciertas cargas y, sin embargo, soportar otras. Finalmente, se aplica toda la metodología planteada en simular y analizar un posible incidente en vuelo, el fenómeno de la pérdida de pala de un turbohélice. Se trata de un suceso muy particular en el que la estructura tiene que soportar unas cargas complejas y excepcionales con las que la aeronave debe ser capaz de completar con éxito el vuelo. El análisis incluye comportamientos no lineales, daños, y varios tipos de fallos, y en el que se trata de identificar los parámetros clave en la secuencia del fallo. El suceso se analiza mediante análisis estructurales deterministas más habituales y también mediante otras técnicas como el método de Monte Carlo con el que se logran estudiar distintas incertidumbres en los parámetros con variables aleatorias. Se estudian, entre otros, el tamaño de pala perdida, la velocidad y el momento en el que se produce la rotura, y la rigidez y resistencia de los apoyos del motor. Se tiene en cuenta incluso el amortiguamiento estructural del sistema. Las distintas estrategias de análisis permiten obtener unos resultados valiosos e interesantes que han sido objeto de distintas publicaciones. ABSTRACT Dynamic phenomena can endanger the integrity of aerospace structures and, consequently, engineers have developed different strategies to analyze them. One of the major engineering problems is how to deal with the structural dynamics. In this thesis, different dynamic phenomena are introduced and several methods are proposed to estimate or simulate their behaviors. The analysis is considered through parametric, deterministic and statistical methods. The suggested issues are from simple problems with few degrees of freedom, in order to develop different strategies and tools to solve them, to very dynamic phenomena containing nonlinear behaviors failures, damages. The first examples cover a wide variety of dynamic phenomena such as vibration analysis of mass elements, including impacts and contacts, and beam analysis with harmonic load applied, in which random parameters are included. These parameters can represent the unawareness or uncertainty of certain variables. During the development of the thesis several concepts are introduced and different methods are applied, such as the finite element method (FEM), which is solved through implicit and explicit schemes, and parametrical and statistical methods using the Monte Carlo analysis technique. Next, once the tools and strategies of analysis are set out more complex phenomena are studied. This is the case of a low-speed impact in composite materials, the residual strength of the structure is evaluated, and therefore, its damage tolerance. This incident may occur from a tool dropped, hail or debris throw on the runway. At an airport may also occur, and it is also analyzed, a collision between an airplane and a frangible device. The devise must brake under these loads, however, it must withstand others. Finally, all the considered methodology is applied to simulate and analyze a flight incident, the blade loss phenomenon of a turboprop. In this particular event the structure must support complex and exceptional loads and the aircraft must be able to successfully complete the flight. Nonlinear behavior, damage, and different types of failures are included in the analysis, in which the key parameters in the failure sequence are identified. The incident is analyzed by deterministic structural analysis and also by other techniques such as Monte Carlo method, in which it is possible to include different parametric uncertainties through random variables. Some of the evaluated parameters are, among others, the blade loss size, propeller rotational frequency, speed and angular position where the blade is lost, and the stiffness and strength of the engine mounts. The study does also research on the structural damping of the system. The different strategies of analysis obtain valuable and interesting results that have been already published.

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A dynamic capsid is critical to the events that shape the viral life cycle; events such as cell attachment, cell entry, and nucleic acid release demand a highly mobile viral surface. Protein mass mapping of the common cold virus, human rhinovirus 14 (HRV14), revealed both viral structural dynamics and the inhibition of such dynamics with an antiviral agent, WIN 52084. Viral capsid digestion fragments resulting from proteolytic time-course experiments provided structural information in good agreement with the HRV14 three-dimensional crystal structure. As expected, initial digestion fragments included peptides from the capsid protein VP1. This observation was expected because VP1 is the most external viral protein. Initial digestion fragments also included peptides belonging to VP4, the most internal capsid protein. The mass spectral results together with x-ray crystallography data provide information consistent with a “breathing” model of the viral capsid. Whereas the crystal structure of HRV14 shows VP4 to be the most internal capsid protein, mass spectral results show VP4 fragments to be among the first digestion fragments observed. Taken together this information demonstrates that VP4 is transiently exposed to the viral surface via viral breathing. Comparative digests of HRV14 in the presence and absence of WIN 52084 revealed a dramatic inhibition of digestion. These results indicate that the binding of the antiviral agent not only causes local conformational changes in the drug binding pocket but actually stabilizes the entire viral capsid against enzymatic degradation. Viral capsid mass mapping provides a fast and sensitive method for probing viral structural dynamics as well as providing a means for investigating antiviral drug efficacy.

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Mutant, but not wild-type p53 binds with high affinity to a variety of MAR-DNA elements (MARs), suggesting that MAR-binding of mutant p53 relates to the dominant-oncogenic activities proposed for mutant p53. MARs recognized by mutant p53 share AT richness and contain variations of an AATATATTT “DNA-unwinding motif,” which enhances the structural dynamics of chromatin and promotes regional DNA base-unpairing. Mutant p53 specifically interacted with MAR-derived oligonucleotides carrying such unwinding motifs, catalyzing DNA strand separation when this motif was located within a structurally labile sequence environment. Addition of GC-clamps to the respective MAR-oligonucleotides or introducing mutations into the unwinding motif strongly reduced DNA strand separation, but supported the formation of tight complexes between mutant p53 and such oligonucleotides. We conclude that the specific interaction of mutant p53 with regions of MAR-DNA with a high potential for base-unpairing provides the basis for the high-affinity binding of mutant p53 to MAR-DNA.