Design and optimization of optical, fiber based PSK demodulators for high-bit-rate optical networks

El objetivo principal de esta tesis ha sido el diseño y la optimización de receptores implementados con fibra óptica, para ser usados en redes ópticas de alta velocidad que empleen formatos de modulación de fase. En los últimos años, los formatos de modulación de fase (Phase Shift keying, PSK) han captado gran atención debido a la mejora de sus prestaciones respecto a los formatos de modulación convencionales. Principalmente, presentan una mejora de la eficiencia espectral y una mayor tolerancia a la degradación de la señal causada por la dispersión cromática, la dispersión por modo de polarización y los efectos no-lineales en la fibra óptica. En este trabajo, se analizan en detalle los formatos PSK, incluyendo sus variantes de modulación de fase diferencial (Differential Phase Shift Keying, DPSK), en cuadratura (Differential Quadrature Phase Shift Keying, DQPSK) y multiplexación en polarización (Polarization Multiplexing Differential Quadrature Phase Shift Keying, PM-DQPSK), con la finalidad de diseñar y optimizar los receptores que permita su demodulación. Para ello, se han analizado y desarrollado nuevas estructuras que ofrecen una mejora en las prestaciones del receptor y una reducción de coste comparadas con las actualmente disponibles. Para la demodulación de señales DPSK, en esta tesis, se proponen dos nuevos receptores basados en un interferómetro en línea Mach-Zehnder (MZI) implementado con tecnología todo-fibra. El principio de funcionamiento de los MZI todo-fibra propuestos se asienta en la interferencia modal que se produce en una fibra multimodo (MMF) cuando se situada entre dos monomodo (SMF). Este tipo de configuración (monomodo-multimodo-monomodo, SMS) presenta un buen ratio de extinción interferente si la potencia acoplada en la fibra multimodo se reparte, principal y equitativamente, entre dos modos dominantes. Con este objetivo, se han estudiado y demostrado tanto teórica como experimentalmente dos nuevas estructuras SMS que mejoran el ratio de extinción. Una de las propuestas se basa en emplear una fibra multimodo de índice gradual cuyo perfil del índice de refracción presenta un hundimiento en su zona central. La otra consiste en una estructura SMS con las fibras desalineadas y donde la fibra multimodo es una fibra de índice gradual convencional. Para las dos estructuras, mediante el análisis teórico desarrollado, se ha demostrado que el 80 – 90% de la potencia de entrada se acopla a los dos modos dominantes de la fibra multimodo y se consigue una diferencia inferior al 10% entre ellos. También se ha demostrado experimentalmente que se puede obtener un ratio de extinción de al menos 12 dB. Con el objeto de demostrar la capacidad de estas estructuras para ser empleadas como demoduladores de señales DPSK, se han realizado numerosas simulaciones de un sistema de transmisión óptico completo y se ha analizado la calidad del receptor bajo diferentes perspectivas, tales como la sensibilidad, la tolerancia a un filtrado óptico severo o la tolerancia a las dispersiones cromática y por modo de polarización. En todos los casos se ha concluido que los receptores propuestos presentan rendimientos comparables a los obtenidos con receptores convencionales. En esta tesis, también se presenta un diseño alternativo para la implementación de un receptor DQPSK, basado en el uso de una fibra mantenedora de la polarización (PMF). A través del análisi teórico y del desarrollo de simulaciones numéricas, se ha demostrado que el receptor DQPSK propuesto presenta prestaciones similares a los convencionales. Para complementar el trabajo realizado sobre el receptor DQPSK basado en PMF, se ha extendido el estudio de su principio de demodulación con el objeto de demodular señales PM-DQPSK, obteniendo como resultado la propuesta de una nueva estructura de demodulación. El receptor PM-DQPSK propuesto se basa en la estructura conjunta de una única línea de retardo junto con un rotador de polarización. Se ha analizado la calidad de los receptores DQPSK y PM-DQPSK bajo diferentes perspectivas, tales como la sensibilidad, la tolerancia a un filtrado óptico severo, la tolerancia a las dispersiones cromática y por modo de polarización o su comportamiento bajo condiciones no-ideales. En comparación con los receptores convencionales, nuestra propuesta exhibe prestaciones similares y además permite un diseño más simple que redunda en un coste potencialmente menor. En las redes de comunicaciones ópticas actuales se utiliza la tecnología de multimplexación en longitud de onda (WDM) que obliga al uso de filtros ópticos con bandas de paso lo más estrechas posibles y a emplear una serie de dispositivos que incorporan filtros en su arquitectura, tales como los multiplexores, demultiplexores, ROADMs, conmutadores y OXCs. Todos estos dispositivos conectados entre sí son equivalentes a una cadena de filtros cuyo ancho de banda se va haciendo cada vez más estrecho, llegando a distorsionar la forma de onda de las señales. Por esto, además de analizar el impacto del filtrado óptico en las señales de 40 Gbps DQPSK y 100 Gbps PM-DQPSK, este trabajo de tesis se completa estudiando qué tipo de filtro óptico minimiza las degradaciones causadas en la señal y analizando el número máximo de filtros concatenados que permiten mantener la calidad requerida al sistema. Se han estudiado y simulado cuatro tipos de filtros ópticos;Butterworth, Bessel, FBG y F-P. ABSTRACT The objective of this thesis is the design and optimization of optical fiber-based phase shift keying (PSK) demodulators for high-bit-rate optical networks. PSK modulation formats have attracted significant attention in recent years, because of the better performance with respect to conventional modulation formats. Principally, PSK signals can improve spectrum efficiency and tolerate more signal degradation caused by chromatic dispersion, polarization mode dispersion and nonlinearities in the fiber. In this work, many PSK formats were analyzed in detail, including the variants of differential phase modulation (Differential Phase Shift Keying, DPSK), in quadrature (Differential Quadrature Phase Shift Keying, DQPSK) and polarization multiplexing (Polarization Multiplexing Differential Quadrature Phase Shift Keying, PM-DQPSK), in order to design and optimize receivers enabling demodulations. Therefore, novel structures, which offer good receiver performances and a reduction in cost compared to the current structures, have been analyzed and developed. Two novel receivers based on an all-fiber in-line Mach-Zehnder interferometer (MZI) were proposed for DPSK signal demodulation in this thesis. The operating principle of the all-fiber MZI is based on the modal interference that occurs in a multimode fiber (MMF) when it is located between two single-mode fibers (SMFs). This type of configuration (Single-mode-multimode-single-mode, SMS) can provide a good extinction ratio if the incoming power from the SMF could be coupled equally into two dominant modes excited in the MMF. In order to improve the interference extinction ratio, two novel SMS structures have been studied and demonstrated, theoretically and experimentally. One of the two proposed MZIs is based on a graded-index multimode fiber (MMF) with a central dip in the index profile, located between two single-mode fibers (SMFs). The other one is based on a conventional graded-index MMF mismatch spliced between two SMFs. Theoretical analysis has shown that, in these two schemes, 80 – 90% of the incoming power can be coupled into the two dominant modes exited in the MMF, and the power difference between them is only ~10%. Experimental results show that interference extinction ratio of 12 dB could be obtained. In order to demonstrate the capacity of these two structures for use as DPSK signal demodulators, numerical simulations in a completed optical transmission system have been carried out, and the receiver quality has been analyzed under different perspectives, such as sensitivity, tolerance to severe optical filtering or tolerance to chromatic and polarization mode dispersion. In all cases, from the simulation results we can conclude that the two proposed receivers can provide performances comparable to conventional ones. In this thesis, an alternative design for the implementation of a DQPSK receiver, which is based on a polarization maintaining fiber (PMF), was also presented. To complement the work made for the PMF-based DQPSK receiver, the study of the demodulation principle has been extended to demodulate PM-DQPSK signals, resulting in the proposal of a novel demodulation structure. The proposed PM-DQPSK receiver is based on only one delay line and a polarization rotator. The quality of the proposed DQPSK and PM-DQPSK receivers under different perspectives, such as sensitivity, tolerance to severe optical filtering, tolerance to chromatic dispersion and polarization mode dispersion, or behavior under non-ideal conditions. Compared with the conventional receivers, our proposals exhibit similar performances but allow a simpler design which can potentially reduce the cost. The wavelength division multiplexing (WDM) technology used in current optical communications networks requires the use of optical filters with a passband as narrow as possible, and the use of a series of devices that incorporate filters in their architecture, such as multiplexers, demultiplexers, switches, reconfigurable add-drop multiplexers (ROADMs) and optical cross-connects (OXCs). All these devices connected together are equivalent to a chain of filters whose bandwidth becomes increasingly narrow, resulting in distortion to the waveform of the signals. Therefore, in addition to analyzing the impact of optical filtering on signal of 40 Gbps DQPSK and 100 Gbps PM-DQPSK, we study which kind of optical filter minimizes the signal degradation and analyze the maximum number of concatenated filters for maintaining the required quality of the system. Four types of optical filters, including Butterworth, Bessel, FBG and FP, have studied and simulated.

Water and nitrogen footprint in an irrigated crop under mineral and organic fertilization

In order to establish rational nitrogen (N) application and reduce groundwater contamination, a clearer understanding of the N distribution through the growing season and its balance is crucial. Excessive doses of N and/or water applied to fertigated crops involve a substantial risk of aquifer contamination by nitrate; but knowledge of N cycling and availability within the soil could assist in avoiding this excess. In central Spain, the main horticultural fertigated crop is the melon type ?piel de sapo¿ and it is cultivated in vulnerable zones to nitrate pollution (Directive 91/676/CEE). However, until few years ago there were not antecedents related to the optimization of nitrogen fertilization together with irrigation. Water and N footprint are indicators that allow assessing the impact generated by different agricultural practices, so they can be used to improve the management strategies in fertigated crop systems. The water footprint distinguishes between blue water (sources of water applied to the crop, like irrigation and precipitation), green water (water used by the crop and stored in the soil), and it is furthermore possible to quantify the impact of pollution by calculating the grey water, which is defined as the volume of polluted water created from the growing and production of crops. On the other hand, the N footprint considers green N (nitrogen consumed by the crops and stored in the soil), blue N (N available for crop, like N applied with mineral and/or organic fertilizers, N applied with irrigation water and N mineralized during the crop period), whereas grey N is the amount of N-NO3- washed from the soil to the aquifer. All these components are expressed as the ratio between the components of water or N footprint and the yield (m3 t-1 or kg N t-1 respectively). The objetives of this work were to evaluate the impact derivated from the use of different fertilizer practices in a melon crop using water and N footprint.

A methodology to analyze, design and implement very fast and robust controls of Buck-type converters

La electrónica digital moderna presenta un desafío a los diseñadores de sistemas de potencia. El creciente alto rendimiento de microprocesadores, FPGAs y ASICs necesitan sistemas de alimentación que cumplan con requirimientos dinámicos y estáticos muy estrictos. Específicamente, estas alimentaciones son convertidores DC-DC de baja tensión y alta corriente que necesitan ser diseñados para tener un pequeño rizado de tensión y una pequeña desviación de tensión de salida bajo transitorios de carga de una alta pendiente. Además, dependiendo de la aplicación, se necesita cumplir con otros requerimientos tal y como proveer a la carga con ”Escalado dinámico de tensión”, donde el convertidor necesitar cambiar su tensión de salida tan rápidamente posible sin sobreoscilaciones, o ”Posicionado Adaptativo de la Tensión” donde la tensión de salida se reduce ligeramente cuanto más grande sea la potencia de salida. Por supuesto, desde el punto de vista de la industria, las figuras de mérito de estos convertidores son el coste, la eficiencia y el tamaño/peso. Idealmente, la industria necesita un convertidor que es más barato, más eficiente, más pequeño y que aún así cumpla con los requerimienos dinámicos de la aplicación. En este contexto, varios enfoques para mejorar la figuras de mérito de estos convertidores se han seguido por la industria y la academia tales como mejorar la topología del convertidor, mejorar la tecnología de semiconducores y mejorar el control. En efecto, el control es una parte fundamental en estas aplicaciones ya que un control muy rápido hace que sea más fácil que una determinada topología cumpla con los estrictos requerimientos dinámicos y, consecuentemente, le da al diseñador un margen de libertar más amplio para mejorar el coste, la eficiencia y/o el tamaño del sistema de potencia. En esta tesis, se investiga cómo diseñar e implementar controles muy rápidos para el convertidor tipo Buck. En esta tesis se demuestra que medir la tensión de salida es todo lo que se necesita para lograr una respuesta casi óptima y se propone una guía de diseño unificada para controles que sólo miden la tensión de salida Luego, para asegurar robustez en controles muy rápidos, se proponen un modelado y un análisis de estabilidad muy precisos de convertidores DC-DC que tienen en cuenta circuitería para sensado y elementos parásitos críticos. También, usando este modelado, se propone una algoritmo de optimización que tiene en cuenta las tolerancias de los componentes y sensados distorsionados. Us ando este algoritmo, se comparan controles muy rápidos del estado del arte y su capacidad para lograr una rápida respuesta dinámica se posiciona según el condensador de salida utilizado. Además, se propone una técnica para mejorar la respuesta dinámica de los controladores. Todas las propuestas se han corroborado por extensas simulaciones y prototipos experimentales. Con todo, esta tesis sirve como una metodología para ingenieros para diseñar e implementar controles rápidos y robustos de convertidores tipo Buck. ABSTRACT Modern digital electronics present a challenge to designers of power systems. The increasingly high-performance of microprocessors, FPGAs (Field Programmable Gate Array) and ASICs (Application-Specific Integrated Circuit) require power supplies to comply with very demanding static and dynamic requirements. Specifically, these power supplies are low-voltage/high-current DC-DC converters that need to be designed to exhibit low voltage ripple and low voltage deviation under high slew-rate load transients. Additionally, depending on the application, other requirements need to be met such as to provide to the load ”Dynamic Voltage Scaling” (DVS), where the converter needs to change the output voltage as fast as possible without underdamping, or ”Adaptive Voltage Positioning” (AVP) where the output voltage is slightly reduced the greater the output power. Of course, from the point of view of the industry, the figures of merit of these converters are the cost, efficiency and size/weight. Ideally, the industry needs a converter that is cheaper, more efficient, smaller and that can still meet the dynamic requirements of the application. In this context, several approaches to improve the figures of merit of these power supplies are followed in the industry and academia such as improving the topology of the converter, improving the semiconductor technology and improving the control. Indeed, the control is a fundamental part in these applications as a very fast control makes it easier for the topology to comply with the strict dynamic requirements and, consequently, gives the designer a larger margin of freedom to improve the cost, efficiency and/or size of the power supply. In this thesis, how to design and implement very fast controls for the Buck converter is investigated. This thesis proves that sensing the output voltage is all that is needed to achieve an almost time-optimal response and a unified design guideline for controls that only sense the output voltage is proposed. Then, in order to assure robustness in very fast controls, a very accurate modeling and stability analysis of DC-DC converters is proposed that takes into account sensing networks and critical parasitic elements. Also, using this modeling approach, an optimization algorithm that takes into account tolerances of components and distorted measurements is proposed. With the use of the algorithm, very fast analog controls of the state-of-the-art are compared and their capabilities to achieve a fast dynamic response are positioned de pending on the output capacitor. Additionally, a technique to improve the dynamic response of controllers is also proposed. All the proposals are corroborated by extensive simulations and experimental prototypes. Overall, this thesis serves as a methodology for engineers to design and implement fast and robust controls for Buck-type converters.

A framework for adaptive e-learning for continuum mechanics and structural analysis

This paper presents a project for providing the students of Structural Engineering with the flexibility to learn outside classroom schedules. The goal is a framework for adaptive E-learning based on a repository of open educational courseware with a set of basic Structural Engineering concepts and fundamentals. These are paramount for students to expand their technical knowledge and skills in structural analysis and design of tall buildings, arch-type structures as well as bridges. Thus, concepts related to structural behaviour such as linearity, compatibility, stiffness and influence lines have traditionally been elusive for students. The objective is to facilitate the student a teachinglearning process to acquire the necessary intuitive knowledge, cognitive skills and the basis for further technological modules and professional development in this area. As a side effect, the system is expected to help the students improve their preparation for exams on the subject. In this project, a web-based open-source system for studying influence lines on continuous beams is presented. It encompasses a collection of interactive user-friendly applications accessible via Web, written in JavaScript under JQuery and Dygraph Libraries, taking advantage of their efficiency and graphic capabilities. It is performed in both Spanish and English languages. The student is enabled to set the geometric, topologic, boundary and mechanic layout of a continuous beam. While changing the loading and the support conditions, the changes in the beam response prompt on the screen, so that the effects of the several issues involved in structural analysis become apparent. This open interaction with the user allows the student to simulate and virtually infer the structural response. Different levels of complexity can be handled, whereas an ongoing help is at hand for any of them. Students can freely boost their experiential learning on this subject at their own pace, in order to further share, process, generalize and apply the relevant essential concepts of Structural Engineering analysis. Besides, this collection is being added to the "Virtual Lab of Continuum Mechanics" of the UPM, launched in 2013 (http://serviciosgate.upm.es/laboratoriosvirtuales/laboratorios/medios-continuos-en-construcci%C3%B3n)

Monte Carlo simulations of densely-packed athermal polymers in the bulk and under confinement

We review the main results from extensive Monte Carlo (MC) simulations on athermal polymer packings in the bulk and under confinement. By employing the simplest possible model of excluded volume, macromolecules are represented as freely-jointed chains of hard spheres of uniform size. Simulations are carried out in a wide concentration range: from very dilute up to very high volume fractions, reaching the maximally random jammed (MRJ) state. We study how factors like chain length, volume fraction and flexibility of bond lengths affect the structure, shape and size of polymers, their packing efficiency and their phase behaviour (disorder–order transition). In addition, we observe how these properties are affected by confinement realized by flat, impenetrable walls in one dimension. Finally, by mapping the parent polymer chains to primitive paths through direct geometrical algorithms, we analyse the characteristics of the entanglement network as a function of packing density.

Biometric authentication of users through iris by using key binding and similarity preserving hash functions = Autenticación biométrica de usuarios a través del iris mediante la ocultación de claves y funciones resumen que preservan la similitud

El extraordinario auge de las nuevas tecnologías de la información, el desarrollo de la Internet de las Cosas, el comercio electrónico, las redes sociales, la telefonía móvil y la computación y almacenamiento en la nube, han proporcionado grandes beneficios en todos los ámbitos de la sociedad. Junto a éstos, se presentan nuevos retos para la protección y privacidad de la información y su contenido, como la suplantación de personalidad y la pérdida de la confidencialidad e integridad de los documentos o las comunicaciones electrónicas. Este hecho puede verse agravado por la falta de una frontera clara que delimite el mundo personal del mundo laboral en cuanto al acceso de la información. En todos estos campos de la actividad personal y laboral, la Criptografía ha jugado un papel fundamental aportando las herramientas necesarias para garantizar la confidencialidad, integridad y disponibilidad tanto de la privacidad de los datos personales como de la información. Por otro lado, la Biometría ha propuesto y ofrecido diferentes técnicas con el fin de garantizar la autentificación de individuos a través del uso de determinadas características personales como las huellas dáctilares, el iris, la geometría de la mano, la voz, la forma de caminar, etc. Cada una de estas dos ciencias, Criptografía y Biometría, aportan soluciones a campos específicos de la protección de datos y autentificación de usuarios, que se verían enormemente potenciados si determinadas características de ambas ciencias se unieran con vistas a objetivos comunes. Por ello es imperativo intensificar la investigación en estos ámbitos combinando los algoritmos y primitivas matemáticas de la Criptografía con la Biometría para dar respuesta a la demanda creciente de nuevas soluciones más técnicas, seguras y fáciles de usar que potencien de modo simultáneo la protección de datos y la identificacíón de usuarios. En esta combinación el concepto de biometría cancelable ha supuesto una piedra angular en el proceso de autentificación e identificación de usuarios al proporcionar propiedades de revocación y cancelación a los ragos biométricos. La contribución de esta tesis se basa en el principal aspecto de la Biometría, es decir, la autentificación segura y eficiente de usuarios a través de sus rasgos biométricos, utilizando tres aproximaciones distintas: 1. Diseño de un esquema criptobiométrico borroso que implemente los principios de la biometría cancelable para identificar usuarios lidiando con los problemas acaecidos de la variabilidad intra e inter-usuarios. 2. Diseño de una nueva función hash que preserva la similitud (SPHF por sus siglas en inglés). Actualmente estas funciones se usan en el campo del análisis forense digital con el objetivo de buscar similitudes en el contenido de archivos distintos pero similares de modo que se pueda precisar hasta qué punto estos archivos pudieran ser considerados iguales. La función definida en este trabajo de investigación, además de mejorar los resultados de las principales funciones desarrolladas hasta el momento, intenta extender su uso a la comparación entre patrones de iris. 3. Desarrollando un nuevo mecanismo de comparación de patrones de iris que considera tales patrones como si fueran señales para compararlos posteriormente utilizando la transformada de Walsh-Hadarmard. Los resultados obtenidos son excelentes teniendo en cuenta los requerimientos de seguridad y privacidad mencionados anteriormente. Cada uno de los tres esquemas diseñados han sido implementados para poder realizar experimentos y probar su eficacia operativa en escenarios que simulan situaciones reales: El esquema criptobiométrico borroso y la función SPHF han sido implementados en lenguaje Java mientras que el proceso basado en la transformada de Walsh-Hadamard en Matlab. En los experimentos se ha utilizado una base de datos de imágenes de iris (CASIA) para simular una población de usuarios del sistema. En el caso particular de la función de SPHF, además se han realizado experimentos para comprobar su utilidad en el campo de análisis forense comparando archivos e imágenes con contenido similar y distinto. En este sentido, para cada uno de los esquemas se han calculado los ratios de falso negativo y falso positivo. ABSTRACT The extraordinary increase of new information technologies, the development of Internet of Things, the electronic commerce, the social networks, mobile or smart telephony and cloud computing and storage, have provided great benefits in all areas of society. Besides this fact, there are new challenges for the protection and privacy of information and its content, such as the loss of confidentiality and integrity of electronic documents and communications. This is exarcebated by the lack of a clear boundary between the personal world and the business world as their differences are becoming narrower. In both worlds, i.e the personal and the business one, Cryptography has played a key role by providing the necessary tools to ensure the confidentiality, integrity and availability both of the privacy of the personal data and information. On the other hand, Biometrics has offered and proposed different techniques with the aim to assure the authentication of individuals through their biometric traits, such as fingerprints, iris, hand geometry, voice, gait, etc. Each of these sciences, Cryptography and Biometrics, provides tools to specific problems of the data protection and user authentication, which would be widely strengthen if determined characteristics of both sciences would be combined in order to achieve common objectives. Therefore, it is imperative to intensify the research in this area by combining the basics mathematical algorithms and primitives of Cryptography with Biometrics to meet the growing demand for more secure and usability techniques which would improve the data protection and the user authentication. In this combination, the use of cancelable biometrics makes a cornerstone in the user authentication and identification process since it provides revocable or cancelation properties to the biometric traits. The contributions in this thesis involve the main aspect of Biometrics, i.e. the secure and efficient authentication of users through their biometric templates, considered from three different approaches. The first one is designing a fuzzy crypto-biometric scheme using the cancelable biometric principles to take advantage of the fuzziness of the biometric templates at the same time that it deals with the intra- and inter-user variability among users without compromising the biometric templates extracted from the legitimate users. The second one is designing a new Similarity Preserving Hash Function (SPHF), currently widely used in the Digital Forensics field to find similarities among different files to calculate their similarity level. The function designed in this research work, besides the fact of improving the results of the two main functions of this field currently in place, it tries to expand its use to the iris template comparison. Finally, the last approach of this thesis is developing a new mechanism of handling the iris templates, considering them as signals, to use the Walsh-Hadamard transform (complemented with three other algorithms) to compare them. The results obtained are excellent taking into account the security and privacy requirements mentioned previously. Every one of the three schemes designed have been implemented to test their operational efficacy in situations that simulate real scenarios: The fuzzy crypto-biometric scheme and the SPHF have been implemented in Java language, while the process based on the Walsh-Hadamard transform in Matlab. The experiments have been performed using a database of iris templates (CASIA-IrisV2) to simulate a user population. The case of the new SPHF designed is special since previous to be applied i to the Biometrics field, it has been also tested to determine its applicability in the Digital Forensic field comparing similar and dissimilar files and images. The ratios of efficiency and effectiveness regarding user authentication, i.e. False Non Match and False Match Rate, for the schemes designed have been calculated with different parameters and cases to analyse their behaviour.

Design and evaluation of a dehumidifying plaster panel for passive architecture integration

Buildings Indoor Air Quality requires a control in the Relative Humidity parameter. In passive architecture in humid climates relative humidity is even more important for human comfort and difficult to control. Therefore, nowadays, there is a research on dehumidifying systems. The present article shows an innovative dehumidifying panel composed of a plaster and Calcium Chloride salt. Laboratory tests are carried out to establish its viability as an indoor air moister regulator integrated in common plaster building interior coatings. There are two types of tests that have been carried out in two consecutive empirical phases: in the first phase, the tests of characterization of the Calcium Chloride as a desiccant are carried out; in a second phase, the dehumidifying panel as a whole is tested. Finally, both types of empirical tests show the efficiency and viability as an air moisture passive control system.

Bio-based polymer nanocomposites based on nylon 11 and WS2 inorganic nanotubes

Tungsten disulphide nanotubes (INT-WS2) have been successfully dispersed in a bio-based polyamide matrix (nylon 11) by conventional melt processing. The effect of INT-WS2 content on the morphology, thermal stability, crystallization behaviour and dynamic mechanical properties is investigated. The results indicate that these inorganic nanotubes can be efficiently incorporated into the bio-based polymer matrix without the need for modifiers or surfactants. Additionally, it is found that the non-isothermal crystallization behaviour of nylon 11/INT-WS2 depends on both the cooling rate and INT-WS2 concentration. In particular, crystallization kinetics results demonstrate that the nucleating activity of INTs plays a dominant role in accelerating the crystallization of nylon 11. This fact leads to the appearance of the more-disordered phase at higher temperature. More significantly, it was shown that these INT-WS2 nanocomposites can facilitate a good processability and cost efficiency, and will be of interest for many eco-friendly and medical applications.

An artificial cytochrome P450 that hydroxylates unactivated carbons with regio- and stereoselectivity and useful catalytic turnovers

A catalyst has been synthesized comprising a manganese porphyrin carrying four beta-cyclodextrin groups. It catalyzes the hydroxylation of substrates of appropriate size carrying tert-butylphenyl groups that can hydrophobically bind into the cyclodextrin cavities. In one example as many as 650 catalytic turnovers are seen before the catalyst is oxidatively destroyed, and with a rate comparable to that of typical cytochrome P450 enzymes. In another example, a steroid derivative is regio- and stereoselectively hydroxylated at a single unactivated carbon atom, but more slowly and with fewer turnovers. The carbon attacked is not the most chemically reactive, and the selectivity is determined by the geometry of the catalyst-substrate complex. Nonbinding substrates are not reactive under the conditions used, and substrates with more flexible binding geometries give more than a single product.

T-strand integration in maize protoplasts after codelivery of a T-DNA substrate and virulence genes

We describe a plant protoplast transformation method that provides transformants with a simple pattern of integration of a foreign gene. The approach is to deliver into plant protoplasts by direct gene transfer the Agrobacterium virulence genes virD1 and virD2 with or without virE2, together with a target plasmid containing a gene of interest flanked by Agrobacterium T-DNA border repeat sequences of 25 bp. We present evidence of T-DNA formation in maize protoplasts and its integration into the maize genome. The frequency of VirD1-VirD2-mediated integration events was about 20–35% of the total number of transformants. The addition of virE2 doubled the transformation efficiency. The method described here is of sufficient efficiency and simplicity to be useful for the production of transgenic plants with single-copy well-defined transgenic inserts.

A link between protein structure and enzyme catalyzed hydrogen tunneling

We present evidence that the size of an active site side chain may modulate the degree of hydrogen tunneling in an enzyme-catalyzed reaction. Primary and secondary kH/kT and kD/kT kinetic isotope effects have been measured for the oxidation of benzyl alcohol catalyzed by horse liver alcohol dehydrogenase at 25°C. As reported in earlier studies, the relationship between secondary kH/kT and kD/kT isotope effects provides a sensitive probe for deviations from classical behavior. In the present work, catalytic efficiency and the extent of hydrogen tunneling have been correlated for the alcohol dehydrogenase-catalyzed hydride transfer among a group of site-directed mutants at position 203. Val-203 interacts with the opposite face of the cofactor NAD+ from the alcohol substrate. The reduction in size of this residue is correlated with diminished tunneling and a two orders of magnitude decrease in catalytic efficiency. Comparison of the x-ray crystal structures of a ternary complex of a high-tunneling (Phe-93 → Trp) and a low-tunneling (Val-203 → Ala) mutant provides a structural basis for the observed effects, demonstrating an increase in the hydrogen transfer distance for the low-tunneling mutant. The Val-203 → Ala ternary complex crystal structure also shows a hyperclosed interdomain geometry relative to the wild-type and the Phe-93 → Trp mutant ternary complex structures. This demonstrates a flexibility in interdomain movement that could potentially narrow the distance between the donor and acceptor carbons in the native enzyme and may enhance the role of tunneling in the hydride transfer reaction.

Organization of G Proteins and Adenylyl Cyclase at the Plasma Membrane

There is mounting evidence for the organization and compartmentation of signaling molecules at the plasma membrane. We find that hormone-sensitive adenylyl cyclase activity is enriched in a subset of regulatory G protein-containing fractions of the plasma membrane. These subfractions resemble, in low buoyant density, structures of the plasma membrane termed caveolae. Immunofluorescence experiments revealed a punctate pattern of G protein α and β subunits, consistent with concentration of these proteins at distinct sites on the plasma membrane. Partial coincidence of localization of G protein α subunits with caveolin (a marker for caveolae) was observed by double immunofluorescence. Results of immunogold electron microscopy suggest that some G protein is associated with invaginated caveolae, but most of the protein resides in irregular structures of the plasma membrane that could not be identified morphologically. Because regulated adenylyl cyclase activity is present in low-density subfractions of plasma membrane from a cell type (S49 lymphoma) that does not express caveolin, this protein is not required for organization of the adenylyl cyclase system. The data suggest that hormone-sensitive adenylyl cyclase systems are localized in a specialized subdomain of the plasma membrane that may optimize the efficiency and fidelity of signal transduction.

Kinetic equilibrium of forces and molecular events in muscle contraction

In biomolecular systems, the mechanical transfer of free energy occurs with both high efficiency and high speed. It is shown here that such a transfer can be achieved only if the participating free-energy-storing elements exhibit opposing relationships between their content of free energy and the force they exert in the transfer direction. A kinetic equilibrium of forces (KEF) results, in which the transfer of free energy is mediated essentially by thermal molecular motion. On the basis of present evidence, KEF is used as a guiding principle in developing a mechanical model of the crossbridge cycle in muscle contraction. The model allows the basic features of molecular events to be visualized in terms of plausible structures. Real understanding of the process will require identification of the elements that perform the functions described here. Besides chemomechanical energy transduction, KEF may have a role in other biomolecular processes in which free energy is transferred mechanically over large distances.

Pressure-mediated oligonucleotide transfection of rat and human cardiovascular tissues

The application of gene therapy to human disease is currently restricted by the relatively low efficiency and potential hazards of methods of oligonucleotide or gene delivery. Antisense or transcription factor decoy oligonucleotides have been shown to be effective at altering gene expression in cell culture expreriments, but their in vivo application is limited by the efficiency of cellular delivery, the intracellular stability of the compounds, and their duration of activity. We report herein the development of a highly efficient method for naked oligodeoxynucleotide (ODN) transfection into cardiovascular tissues by using controlled, nondistending pressure without the use of viral vectors, lipid formulations, or exposure to other adjunctive, potentially hazardous substances. In this study, we have documented the ability of ex vivo, pressure-mediated transfection to achieve nuclear localization of fluorescent (FITC)-labeled ODN in approximately 90% and 50% of cells in intact human saphenous vein and rat myocardium, respectively. We have further documented that pressure-mediated delivery of antisense ODN can functionally inhibit target gene expression in both of these tissues in a sequence-specific manner at the mRNA and protein levels. This oligonucleotide transfection system may represent a safe means of achieving the intraoperative genetic engineering of failure-resistant human bypass grafts and may provide an avenue for the genetic manipultation of cardiac allograft rejection, allograft vasculopathy, or other transplant diseases.

Reconstitution and functional comparison of purified GlpF and AqpZ, the glycerol and water channels from Escherichia coli

A large family of membrane channel proteins selective for transport of water (aquaporins) or water plus glycerol (aquaglyceroporins) has been found in diverse life forms. Escherichia coli has two members of this family—a water channel, AqpZ, and a glycerol facilitator, GlpF. Despite having similar primary amino acid sequences and predicted structures, the oligomeric state and solute selectivity of AqpZ and GlpF are disputed. Here we report biochemical and functional characterizations of affinity-purified GlpF and compare it to AqpZ. Histidine-tagged (His-GlpF) and hemagglutinin-tagged (HA-GlpF) polypeptides encoded by a bicistronic construct were expressed in bacteria. HA-GlpF and His-GlpF appear to form oligomers during Ni-nitrilotriacetate affinity purification. Sucrose gradient sedimentation analyses showed that the oligomeric state of octyl glucoside-solubilized GlpF varies: low ionic strength favors subunit dissociation, whereas Mg2+ stabilizes tetrameric assembly. Reconstitution of affinity-purified GlpF into proteoliposomes increases glycerol permeability more than 100-fold and water permeability up to 10-fold compared with control liposomes. Glycerol and water permeability of GlpF both occur with low Arrhenius activation energies and are reversibly inhibited by HgCl2. Our studies demonstrate that, unlike AqpZ, a water-selective stable tetramer, purified GlpF exists in multiple oligomeric forms under nondenaturing conditions and is highly permeable to glycerol but less well permeated by water.