New methods for quantification and analysis of quantitative real-time polymerase chain reaction data

Quantitative real-time polymerase chain reaction (qPCR) is a sensitive gene quantitation method that has been widely used in the biological and biomedical fields. The currently used methods for PCR data analysis, including the threshold cycle (CT) method, linear and non-linear model fitting methods, all require subtracting background fluorescence. However, the removal of background fluorescence is usually inaccurate, and therefore can distort results. Here, we propose a new method, the taking-difference linear regression method, to overcome this limitation. Briefly, for each two consecutive PCR cycles, we subtracted the fluorescence in the former cycle from that in the later cycle, transforming the n cycle raw data into n-1 cycle data. Then linear regression was applied to the natural logarithm of the transformed data. Finally, amplification efficiencies and the initial DNA molecular numbers were calculated for each PCR run. To evaluate this new method, we compared it in terms of accuracy and precision with the original linear regression method with three background corrections, being the mean of cycles 1-3, the mean of cycles 3-7, and the minimum. Three criteria, including threshold identification, max R2, and max slope, were employed to search for target data points. Considering that PCR data are time series data, we also applied linear mixed models. Collectively, when the threshold identification criterion was applied and when the linear mixed model was adopted, the taking-difference linear regression method was superior as it gave an accurate estimation of initial DNA amount and a reasonable estimation of PCR amplification efficiencies. When the criteria of max R2 and max slope were used, the original linear regression method gave an accurate estimation of initial DNA amount. Overall, the taking-difference linear regression method avoids the error in subtracting an unknown background and thus it is theoretically more accurate and reliable. This method is easy to perform and the taking-difference strategy can be extended to all current methods for qPCR data analysis.^

The introduction of teaching innovations into the traditional teaching of construction and building materials

The traditional teaching methods used for training civil engineers are currently being called into question as a result of the new knowledge and skills now required by the labor market. In addition, the European Higher Education Area is requesting that students be given a greater say in their learning. In the subject called Construction and Building Materials at the Civil Engineering School of the Universidad Politécnica de Madrid, a path was set three academic years ago to lead to an improvement in traditional teaching by introducing active methodologies. The innovations are based on cooperative learning, new technologies, and continuous assessment. The writers’ proposal is to offer their experience as a contribution to the debate on how students can be encouraged to acquire the skills currently demanded from a civil engineer, though not overlooking solid, top-quality training. From the outcomes obtained, it can be concluded that using new teaching techniques to supplement a traditional approach provides more opportunities for students to learn while boosting their motivation. In our case, the introduction of these changes has resulted in an increased pass rate of 29% on average, when such a figure is considered in the light of the mean value of passes during the last decade.

New methods for the estimation of Takagi-Sugeno model based extended Kalman filter and its applications to optimal control for nonlinear systems

This paper describes new approaches to improve the local and global approximation (matching) and modeling capability of Takagi–Sugeno (T-S) fuzzy model. The main aim is obtaining high function approximation accuracy and fast convergence. The main problem encountered is that T-S identification method cannot be applied when the membership functions are overlapped by pairs. This restricts the application of the T-S method because this type of membership function has been widely used during the last 2 decades in the stability, controller design of fuzzy systems and is popular in industrial control applications. The approach developed here can be considered as a generalized version of T-S identification method with optimized performance in approximating nonlinear functions. We propose a noniterative method through weighting of parameters approach and an iterative algorithm by applying the extended Kalman filter, based on the same idea of parameters’ weighting. We show that the Kalman filter is an effective tool in the identification of T-S fuzzy model. A fuzzy controller based linear quadratic regulator is proposed in order to show the effectiveness of the estimation method developed here in control applications. An illustrative example of an inverted pendulum is chosen to evaluate the robustness and remarkable performance of the proposed method locally and globally in comparison with the original T-S model. Simulation results indicate the potential, simplicity, and generality of the algorithm. An illustrative example is chosen to evaluate the robustness. In this paper, we prove that these algorithms converge very fast, thereby making them very practical to use.

Process of Expropriation of Buildings for the Construction of the New Dock of the Port of Cartagena (Spain) in the 18th Century

The need of the Bourbon monarchy to build a Naval Base in the Bay of Cartagena (Spain) during the eighteenth century, implied performing various actions on the environment which allowed the construction of the new dock. One of the priority actions was the transformation of the watershed of the streams that flowed into Mandaraches´s sea. For this reason, a dike was designed and constructed in the northern part of the city. The design of this great work, which was designed as a fortification of the city, was subject to considerable uncertainties. Its proximity to the city involved the demolition of several buildings in the San Roque´s neighborhood. The greater or lesser number of affected buildings and the value of the just indemnification for the expropriation of them, become decisive factors to determine if the work was viable for the Royal Estate or not.

New methods for the synthesis of radiation patterns of coupled antenna arrays = Nuevos métodos de síntesis de diagramas de radiación de agrupaciones de antenas acopladas

El principal objetivo de esta tesis es el desarrollo de métodos de síntesis de diagramas de radiación de agrupaciones de antenas, en donde se realiza una caracterización electromagnética rigurosa de los elementos radiantes y de los acoplos mutuos existentes. Esta caracterización no se realiza habitualmente en la gran mayoría de métodos de síntesis encontrados en la literatura, debido fundamentalmente a dos razones. Por un lado, se considera que el diagrama de radiación de un array de antenas se puede aproximar con el factor de array que únicamente tiene en cuenta la posición de los elementos y las excitaciones aplicadas a los mismos. Sin embargo, como se mostrará en esta tesis, en múltiples ocasiones un riguroso análisis de los elementos radiantes y del acoplo mutuo entre ellos es importante ya que los resultados obtenidos pueden ser notablemente diferentes. Por otro lado, no es sencillo combinar un método de análisis electromagnético con un proceso de síntesis de diagramas de radiación. Los métodos de análisis de agrupaciones de antenas suelen ser costosos computacionalmente, ya que son estructuras grandes en términos de longitudes de onda. Generalmente, un diseño de un problema electromagnético suele comprender varios análisis de la estructura, dependiendo de las variaciones de las características, lo que hace este proceso muy costoso. Dos métodos se utilizan en esta tesis para el análisis de los arrays acoplados. Ambos están basados en el método de los elementos finitos, la descomposición de dominio y el análisis modal para analizar la estructura radiante y han sido desarrollados en el grupo de investigación donde se engloba esta tesis. El primero de ellos es una técnica de análisis de arrays finitos basado en la aproximación de array infinito. Su uso es indicado para arrays planos de grandes dimensiones con elementos equiespaciados. El segundo caracteriza el array y el acoplo mutuo entre elementos a partir de una expansión en modos esféricos del campo radiado por cada uno de los elementos. Este método calcula los acoplos entre los diferentes elementos del array usando las propiedades de traslación y rotación de los modos esféricos. Es capaz de analizar agrupaciones de elementos distribuidos de forma arbitraria. Ambas técnicas utilizan una formulación matricial que caracteriza de forma rigurosa el campo radiado por el array. Esto las hace muy apropiadas para su posterior uso en una herramienta de diseño, como los métodos de síntesis desarrollados en esta tesis. Los resultados obtenidos por estas técnicas de síntesis, que incluyen métodos rigurosos de análisis, son consecuentemente más precisos. La síntesis de arrays consiste en modificar uno o varios parámetros de las agrupaciones de antenas buscando unas determinadas especificaciones de las características de radiación. Los parámetros utilizados como variables de optimización pueden ser varios. Los más utilizados son las excitaciones aplicadas a los elementos, pero también es posible modificar otros parámetros de diseño como son las posiciones de los elementos o las rotaciones de estos. Los objetivos de las síntesis pueden ser dirigir el haz o haces en una determinada dirección o conformar el haz con formas arbitrarias. Además, es posible minimizar el nivel de los lóbulos secundarios o del rizado en las regiones deseadas, imponer nulos que evitan posibles interferencias o reducir el nivel de la componente contrapolar. El método para el análisis de arrays finitos basado en la aproximación de array infinito considera un array finito como un array infinito con un número finito de elementos excitados. Los elementos no excitados están físicamente presentes y pueden presentar tres diferentes terminaciones, corto-circuito, circuito abierto y adaptados. Cada una de estas terminaciones simulará mejor el entorno real en el que el array se encuentre. Este método de análisis se integra en la tesis con dos métodos diferentes de síntesis de diagramas de radiación. En el primero de ellos se presenta un método basado en programación lineal en donde es posible dirigir el haz o haces, en la dirección deseada, además de ejercer un control sobre los lóbulos secundarios o imponer nulos. Este método es muy eficiente y obtiene soluciones óptimas. El mismo método de análisis es también aplicado a un método de conformación de haz, en donde un problema originalmente no convexo (y de difícil solución) es transformado en un problema convexo imponiendo restricciones de simetría, resolviendo de este modo eficientemente un problema complejo. Con este método es posible diseñar diagramas de radiación con haces de forma arbitraria, ejerciendo un control en el rizado del lóbulo principal, así como en el nivel de los lóbulos secundarios. El método de análisis de arrays basado en la expansión en modos esféricos se integra en la tesis con tres técnicas de síntesis de diagramas de radiación. Se propone inicialmente una síntesis de conformación del haz basado en el método de la recuperación de fase resuelta de forma iterativa mediante métodos convexos, en donde relajando las restricciones del problema original se consiguen unas soluciones cercanas a las óptimas de manera eficiente. Dos métodos de síntesis se han propuesto, donde las variables de optimización son las posiciones y las rotaciones de los elementos respectivamente. Se define una función de coste basada en la intensidad de radiación, la cual es minimizada de forma iterativa con el método del gradiente. Ambos métodos reducen el nivel de los lóbulos secundarios minimizando una función de coste. El gradiente de la función de coste es obtenido en términos de la variable de optimización en cada método. Esta función de coste está formada por la expresión rigurosa de la intensidad de radiación y por una función de peso definida por el usuario para imponer prioridades sobre las diferentes regiones de radiación, si así se desea. Por último, se presenta un método en el cual, mediante técnicas de programación entera, se buscan las fases discretas que generan un diagrama de radiación lo más cercano posible al deseado. Con este método se obtienen diseños que minimizan el coste de fabricación. En cada uno de las diferentes técnicas propuestas en la tesis, se presentan resultados con elementos reales que muestran las capacidades y posibilidades que los métodos ofrecen. Se comparan los resultados con otros métodos disponibles en la literatura. Se muestra la importancia de tener en cuenta los diagramas de los elementos reales y los acoplos mutuos en el proceso de síntesis y se comparan los resultados obtenidos con herramientas de software comerciales. ABSTRACT The main objective of this thesis is the development of optimization methods for the radiation pattern synthesis of array antennas in which a rigorous electromagnetic characterization of the radiators and the mutual coupling between them is performed. The electromagnetic characterization is usually overlooked in most of the available synthesis methods in the literature, this is mainly due to two reasons. On the one hand, it is argued that the radiation pattern of an array is mainly influenced by the array factor and that the mutual coupling plays a minor role. As it is shown in this thesis, the mutual coupling and the rigorous characterization of the array antenna influences significantly in the array performance and its computation leads to differences in the results obtained. On the other hand, it is difficult to introduce an analysis procedure into a synthesis technique. The analysis of array antennas is generally expensive computationally as the structure to analyze is large in terms of wavelengths. A synthesis method requires to carry out a large number of analysis, this makes the synthesis problem very expensive computationally or intractable in some cases. Two methods have been used in this thesis for the analysis of coupled antenna arrays, both of them have been developed in the research group in which this thesis is involved. They are based on the finite element method (FEM), the domain decomposition and the modal analysis. The first one obtains a finite array characterization with the results obtained from the infinite array approach. It is specially indicated for the analysis of large arrays with equispaced elements. The second one characterizes the array elements and the mutual coupling between them with a spherical wave expansion of the radiated field by each element. The mutual coupling is computed using the properties of translation and rotation of spherical waves. This method is able to analyze arrays with elements placed on an arbitrary distribution. Both techniques provide a matrix formulation that makes them very suitable for being integrated in synthesis techniques, the results obtained from these synthesis methods will be very accurate. The array synthesis stands for the modification of one or several array parameters looking for some desired specifications of the radiation pattern. The array parameters used as optimization variables are usually the excitation weights applied to the array elements, but some other array characteristics can be used as well, such as the array elements positions or rotations. The desired specifications may be to steer the beam towards any specific direction or to generate shaped beams with arbitrary geometry. Further characteristics can be handled as well, such as minimize the side lobe level in some other radiating regions, to minimize the ripple of the shaped beam, to take control over the cross-polar component or to impose nulls on the radiation pattern to avoid possible interferences from specific directions. The analysis method based on the infinite array approach considers an infinite array with a finite number of excited elements. The infinite non-excited elements are physically present and may have three different terminations, short-circuit, open circuit and match terminated. Each of this terminations is a better simulation for the real environment of the array. This method is used in this thesis for the development of two synthesis methods. In the first one, a multi-objective radiation pattern synthesis is presented, in which it is possible to steer the beam or beams in desired directions, minimizing the side lobe level and with the possibility of imposing nulls in the radiation pattern. This method is very efficient and obtains optimal solutions as it is based on convex programming. The same analysis method is used in a shaped beam technique in which an originally non-convex problem is transformed into a convex one applying symmetry restrictions, thus solving a complex problem in an efficient way. This method allows the synthesis of shaped beam radiation patterns controlling the ripple in the mainlobe and the side lobe level. The analysis method based on the spherical wave expansion is applied for different synthesis techniques of the radiation pattern of coupled arrays. A shaped beam synthesis is presented, in which a convex formulation is proposed based on the phase retrieval method. In this technique, an originally non-convex problem is solved using a relaxation and solving a convex problems iteratively. Two methods are proposed based on the gradient method. A cost function is defined involving the radiation intensity of the coupled array and a weighting function that provides more degrees of freedom to the designer. The gradient of the cost function is computed with respect to the positions in one of them and the rotations of the elements in the second one. The elements are moved or rotated iteratively following the results of the gradient. A highly non-convex problem is solved very efficiently, obtaining very good results that are dependent on the starting point. Finally, an optimization method is presented where discrete digital phases are synthesized providing a radiation pattern as close as possible to the desired one. The problem is solved using linear integer programming procedures obtaining array designs that greatly reduce the fabrication costs. Results are provided for every method showing the capabilities that the above mentioned methods offer. The results obtained are compared with available methods in the literature. The importance of introducing a rigorous analysis into the synthesis method is emphasized and the results obtained are compared with a commercial software, showing good agreement.

Simplified design methods of treated timber structures for shore, beach, and marina construction /

"March 1976."

The new metal worker pattern book; a complete course of instruction in the modern methods of developing and cutting the patterns for sheet metal work, giving the principles underlying practically every problem that is likely to come up in practice and explaining the selection and use of drawing tools and linear and geometrical drawing so clearly that one, who has had no previous knowledge of arithmetic or drawing, may understand these essentials and apply them in using the 253 problems in the development of patterns by the parallel line, conical or flaring, and triangulation systems.

Mode of access: Internet.

A new class of digital division methods /

"This work was supported in part by the Atomic Energy Commission and the Office of Naval Research under AEC Contract AT(11-1)-415."

Experimental essays on medical and philosophical subjects : particularly, I. On the fermentation of alimentary mixtures, and digestion of the food. II. On the nature and properties of fixed air. III. On the respective powers, and manner of acting, of the different kinds of antiseptics. IV. On the scurvy ; with a proposal for trying new methods to prevent or cure the same, at sea. V. On the dissolvent power of quick-lime ; and a further investigation of the properties of fixed air /

Mode of access: Internet.

How to succeed with bees: how fall management saves spring labor, increases next year's honey crop, reduces winter losses far below U.S. average, how bees store more honey when swarming is controlled, successful methods of producing comb or extracted honey, how to make increase with minimum crop loss, how to introduce new queen bees. More than 190 successful plans to produce big crops of honey.

On cover: By E. W. Atkins [and] K. Hawkins.

Methods of preparing horizontal construction joints in mass concrete /

Mode of access: Internet.

Concrete roads and their construction; being a description of the concrete roads in the United Kingdom, together with a summary of the experience in this form of construction gained in Australia, Canada, New Zealand and the United States of America.

Mode of access: Internet.