Kryoelektronenmikroskopie von Proteinen: 3D-Struktur (12 A) des Hämocyanins der Schnecke Haliotis

Zusammenfassung:Die Quartärstruktur des respiratorischen Proteins Hämocyanin (Isoform HtH1) aus der marinen Schnecke Haliotis tuberculata wurde vermittels Kryoelektronen-mikroskopie und 3D-Rekonstruktion untersucht. Das Molekül ist zylinderförmig, hat einen Durchmesser von ca. 35 nm und besteht aus einer Zylinderwand und einem internen Kragenkomplex. Dieser wiederum besteht aus einem Collar und einem Arc.Die kryoelektronenmikroskopischen Aufnahmen von in glasartigem Eis fixierten HtH1-Molekülen brachte eine enorme Verbesserung der Anzahl der zur Verfügung stehenden Ansichtswinkel gegenüber den negativkontrastierten Molekülen, die auf Karbonfilm präpariert waren.Die 3D-Rekonstruktion des HtH1 mittels Aufnahmen bei drei verschiedenen Defo-kuswerten verbesserte die Auflösung noch einmal deutlich gegenüber den Rekon-struktionen, die aus Aufnahmen bei einem festen Defokuswert gemacht wurden, und zwar auf 12 Å. Das Molekül besitzt eine D5-Symmetrie.Aus dieser bisher genausten Rekonstruktion eines Molluskenhämocyanins aus EM-Bildern ließen sich folgende neue Strukturdetails ableiten:· Ein Untereinheitendimer konnte als Repeating Unit im Dekamer des HtH1 beschrieben werden.· Das Untereinheitendimer konnte aus der 3D-Dichtekarte isoliert werden. Es be-steht eindeutig aus 16 Massen, die funktionellen Domänen entsprechen. Zwei dieser Massen bilden den Collar, zwei den Arc und 12 das Wandsegment.· Die gegenläufige Anordnung der beiden Untereinheiten innerhalb dieses Unte-reinheitendimers konnten bestätigt und auf zwei Möglichkeiten eingeschränkt werden.· Die Zahl der alternativen Anordnungen der 16 funktionellen Domänen (HtH1-a bis HtH1-h) im Untereinheitendimer konnten von 80 auf 2 eingeengt werden.· Es konnte über molekulares Modellieren mithilfe einer publizierten Kristallstruk-tur eine 3D-Struktur fastatomarer Auflösung der funktionellen Domäne HtH1-g berechnet werden.· Die funktionelle Domäne HtH1-g konnte als Domänenpaar plausibel in die 3D?Dichtekarte des Untereinheitendimers eingepasst werden, und zwar in die beiden Massen des Arc.Aus der elektronenmikroskopisch gewonnenen Dichtekarte wurde mit Hilfe des

Paleo-sea surface temperature calculations in the equatorial east Atlantic

We present two ~270 kyr paleo-sea surface temperature (SST) records from the Equatorial Divergence and the South Equatorial Current derived from Mg/Ca ratios in the planktic foraminifer Globigerinoides sacculifer. The present study suggests that the magnesium signature of G. sacculifer provides a seasonal SST estimate from the upper ~50 m of the water column generated during upwelling in austral low-latitude fall/winter. Common to both down-core records is a glacial-interglacial amplitude of ~3°-3.5°C for the last climatic changes and lower Holocene and glacial oxygen isotope stage 2 temperatures compared with interglacial stage 5.5 and glacial stage 6 temperatures, respectively. The comparison to published SST estimates from alkenones, oxygen isotopes, and foraminiferal transfer function from the same core material pinpoints discrepancies and conformities between methods.

Contribución al desarrollo de técnicas CAD para el diseño de antenas impresas y dispositivos pasivos de microondas basadas en el método de elementos finitos

El objetivo principal de esta tesis es el desarrollo de herramientas numéricas basadas en técnicas de onda completa para el diseño asistido por ordenador (Computer-Aided Design,‘CAD’) de dispositivos de microondas. En este contexto, se desarrolla una herramienta numérica basada en el método de los elementos finitos para el diseño y análisis de antenas impresas mediante algoritmos de optimización. Esta técnica consiste en dividir el análisis de una antena en dos partes. Una parte de análisis 3D que se realiza sólo una vez en cada punto de frecuencia de la banda de funcionamiento donde se sustituye una superficie que contiene la metalización del parche por puertas artificiales. En una segunda parte se inserta entre las puertas artificiales en la estructura 3D la superficie soportando una metalización y se procede un análisis 2D para caracterizar el comportamiento de la antena. La técnica propuesta en esta tesis se puede implementar en un algoritmo de optimización para definir el perfil de la antena que permite conseguir los objetivos del diseño. Se valida experimentalmente dicha técnica empleándola en el diseño de antenas impresas de banda ancha para diferentes aplicaciones mediante la optimización del perfil de los parches. También, se desarrolla en esta tesis un procedimiento basado en el método de descomposición de dominio y el método de los elementos finitos para el diseño de dispositivos pasivos de microonda. Se utiliza este procedimiento en particular para el diseño y sintonía de filtros de microondas. En la primera etapa de su aplicación se divide la estructura que se quiere analizar en subdominios aplicando el método de descomposición de dominio, este proceso permite analizar cada segmento por separado utilizando el método de análisis adecuado dado que suele haber subdominios que se pueden analizar mediante métodos analíticos por lo que el tiempo de análisis es más reducido. Se utilizan métodos numéricos para analizar los subdominios que no se pueden analizar mediante métodos analíticos. En esta tesis, se utiliza el método de los elementos finitos para llevar a cabo el análisis. Además de la descomposición de dominio, se aplica un proceso de barrido en frecuencia para reducir los tiempos del análisis. Como método de orden reducido se utiliza la técnica de bases reducidas. Se ha utilizado este procedimiento para diseñar y sintonizar varios ejemplos de filtros con el fin de comprobar la validez de dicho procedimiento. Los resultados obtenidos demuestran la utilidad de este procedimiento y confirman su rigurosidad, precisión y eficiencia en el diseño de filtros de microondas. ABSTRACT The main objective of this thesis is the development of numerical tools based on full-wave techniques for computer-aided design ‘CAD’ of microwave devices. In this context, a numerical technique based on the finite element method ‘FEM’ for the design and analysis of printed antennas using optimization algorithms has been developed. The proposed technique consists in dividing the analysis of the antenna in two stages. In the first stage, the regions of the antenna which do not need to be modified during the CAD process are initially characterized only once from their corresponding matrix transfer function (Generalized Admittance matrix, ‘GAM’). The regions which will be modified are defined as artificial ports, precisely the regions which will contain the conducting surfaces of the printed antenna. In a second stage, the contour shape of the conducting surfaces of the printed antenna is iteratively modified in order to achieve a desired electromagnetic performance of the antenna. In this way, a new GAM of the radiating device which takes into account each printed antenna shape is computed after each iteration. The proposed technique can be implemented with a genetic algorithm to achieve the design objectives. This technique is validated experimentally and applied to the design of wideband printed antennas for different applications by optimizing the shape of the radiating device. In addition, a procedure based on the domain decomposition method and the finite element method has been developed for the design of microwave passive devices. In particular, this procedure can be applied to the design and tune of microwave filters. In the first stage of its implementation, the structure to be analyzed is divided into subdomains using the domain decomposition method; this process allows each subdomains can be analyzed separately using suitable analysis method, since there is usually subdomains that can be analyzed by analytical methods so that the time of analysis is reduced. For analyzing the subdomains that cannot be analyzed by analytical methods, we use the numerical methods. In this thesis, the FEM is used to carry out the analysis. Furthermore the decomposition of the domain, a frequency sweep process is applied to reduce analysis times. The reduced order model as the reduced basis technique is used in this procedure. This procedure is applied to the design and tune of several examples of microwave filters in order to check its validity. The obtained results allow concluding the usefulness of this procedure and confirming their thoroughness, accuracy and efficiency for the design of microwave filters.

Wavelength monitoring with semiconductor laser amplifiers

The semiconductor laser diodes that are typically used in applications of optical communications, when working as amplifiers, present under certain conditions optical bistability, which is characterized by abruptly switching between two different output states and an associated hysteresis cycle. This bistable behavior is strongly dependent on the frequency detuning between the frequency of the external optical signal that is injected into the semiconductor laser amplifier and its own emission frequency. This means that small changes in the wavelength of an optical signal applied to a laser amplifier causes relevant changes in the characteristics of its transfer function in terms of the power requirements to achieve bistability and the width of the hysteresis. This strong dependence in the working characteristics of semiconductor laser amplifiers on frequency detuning suggest the use of this kind of devices in optical sensing applications for optical communications, such as the detection of shifts in the emission wavelength of a laser, or detect possible interference between adjacent channels in DWDM (Dense Wavelength Division Multiplexing) optical communication networks

On the harmonic analysis of cup anemometer rotation speed: A principle to monitor performance and maintenance status of rotating meteorological sensors

The calibration results of one anemometer equipped with several rotors, varying their size, were analyzed. In each case, the 30-pulses pert turn output signal of the anemometer was studied using Fourier series decomposition and correlated with the anemometer factor (i.e., the anemometer transfer function). Also, a 3-cup analytical model was correlated to the data resulting from the wind tunnel measurements. Results indicate good correlation between the post-processed output signal and the working condition of the cup anemometer. This correlation was also reflected in the results from the proposed analytical model. With the present work the possibility of remotely checking cup anemometer status, indicating the presence of anomalies and, therefore, a decrease on the wind sensor reliability is revealed.

Caracterización y optimización de estructuras biofotónicas y técnicas avanzadas de interrogación para desarrollo de biosensores ópticos

El núcleo fundamental de esta tesis doctoral es un modelo teórico de la interacción de la luz con un tipo particular de biosensor óptico. Este biosensor se compone de dos regiones: en la región inferior puede haber capas de materiales con diferentes espesores y propiedades ópticas, apiladas horizontalmente; en la zona superior, sobre la que incide directamente el haz de luz, puede haber estructuras que hacen que las propiedades ópticas cambien tanto en el plano horizontal como en la dirección vertical. Estos biosensores responden ópticamente de forma diferente al ser iluminados dependiendo de que su superficie externa esté, en mayor o menor medida, recubierta con diferentes tipos de material biológico. En esta tesis se define un modelo analítico aproximado que permite simular la respuesta óptica de biosensores con estructuras en su región más externa. Una vez comprobada la validez práctica del modelo mediante comparación con medidas experimentales, éste se utiliza en el diseño de biosensores de rendimiento óptimo y en la definición de nuevas técnicas de interrogación óptica. En particular, el sistema de transducción IROP (Increased Relative Optical Power), basado en el efecto que produce la presencia de material biológico, en la potencia total reflejada por la celda biosensora en determinados intervalos espectrales, es uno de los sistemas que ha sido patentado y es objeto de desarrollo por la empresa de base tecnológica BIOD [www.biod.es/], estando ya disponibles en este momento varios dispositivos de diagnóstico basados en esta idea. Los dispositivos basados en este sistema de transducción han demostrado su eficiencia en la detección de proteínas y agentes infecciosos como los rotavirus y el virus del dengue. Finalmente, el modelo teórico desarrollado se utiliza para caracterizar las propiedades ópticas de algunos de los materiales de los que se fabrican los biosensores, así como las de las capas de material biológico formadas en las diferentes fases de un inmunoensayo. Los parámetros ópticos de las capas mencionadas se obtienen mediante el método general de ajuste por mínimos cuadrados a las curvas experimentales obtenidas en los inmunoensayos. ABSTRACT The core of this thesis is the theoretical modeling of the interaction of light with a particular type of optical biosensor. This biosensor consists of two parts: in the lower region may have layers of materials with different thicknesses and optical properties, stacked horizontally; at the top, on which directly affects the light beam, there may be structures that make optical properties change in both, the horizontal and in the vertical direction. These biosensors optically respond differently when illuminated depending on its external surface is greater or lesser extent, coated with different types of biological material. In this thesis an approximate analytical model to simulate the optical response of biosensors with structures in its outer region is defined. After verifying the practical validity of the model by comparison with experimental measurements, it is used in the design of biosensors with optimal performance and the definition of new optical interrogation techniques. In particular, the transduction system IROP (Increased Relative Optical Power) based on the effect of the presence of biological material in the total power reflected from the biosensor cell in certain spectral ranges, has been patented and is under development by the startup company BIOD [www.biod.es/], being already available at this time, several diagnostic devices based on this idea. Devices based on this transduction system have proven their efficiency in detecting proteins and infectious agents such as rotavirus and virus of dengue. Finally, the developed theoretical model is used to characterize the optical properties of some of the materials from which biosensors are fabricated, as well as the optical properties of the biological material layers formed at different stages of an immunoassay. The optical parameters of the layers above are obtained by the general method of least squares fit to the experimental curves obtained in immunoassays.

Radiolarian-based bransfer function for the estimation of sea surface temperatures in the Southern Ocean (Atlantic Sector)

A new radiolarian-based transfer function for sea surface temperature (SST) estimations has been developed from 23 taxa and taxa groups in 53 surface sediment samples recovered between 35° and 72°S in the Atlantic sector of the Southern Ocean. For the selection of taxa and taxa groups ecological information from water column studies was considered. The transfer function allows the estimation of austral summer SST (December-March) ranging between -1 and 18°C with a standard error of estimate of 1.2°C. SST estimates from selected late Pleistocene squences were sucessfully compared with independend paleotemperature estimates derived from a diatom transfer function. This shows that radiolarians provide an excellent tool for paleotemperature reconstructions in Pleistocene sediments of the Southern Ocean.

Three-dimensional solitons in coupled atomic-molecular Bose-Einstein condensates

We present a theoretical analysis of three-dimensional (3D) matter-wave solitons and their stability properties in coupled atomic and molecular Bose-Einstein condensates (BECs). The soliton solutions to the mean-field equations are obtained in an approximate analytical form by means of a variational approach. We investigate soliton stability within the parameter space described by the atom-molecule conversion coupling, the atom-atom s-wave scattering, and the bare formation energy of the molecular species. In terms of ordinary optics, this is analogous to the process of sub- or second-harmonic generation in a quadratic nonlinear medium modified by a cubic nonlinearity, together with a phase mismatch term between the fields. While the possibility of formation of multidimensional spatiotemporal solitons in pure quadratic media has been theoretically demonstrated previously, here we extend this prediction to matter-wave interactions in BEC systems where higher-order nonlinear processes due to interparticle collisions are unavoidable and may not be neglected. The stability of the solitons predicted for repulsive atom-atom interactions is investigated by direct numerical simulations of the equations of motion in a full 3D lattice. Our analysis also leads to a possible technique for demonstrating the ground state of the Schrodinger-Newton and related equations that describe Bose-Einstein condensates with nonlocal interparticle forces.

Applications of ultralong Raman fibre lasers in photonics

This thesis presents a numerical and experimental investigation on applications of ultralong Raman fibre lasers in optical communications, supercontinuum generation and soliton transmission. The research work is divided in four main sections. The first involves the numerical investigation of URFL intra-cavity power and the relative intensity noise transfer evolution along the transmission span. The performance of the URFL is compared with amplification systems of similar complexity. In the case of intracavity power evolution, URFL is compared with a first order Raman amplification system. For the RIN transfer investigation, URFL is compared with a bi-directional dual wavelength pumping system. The RIN transfer function is investigated for several cavity design parameters such as span length, pump distribution and FBG reflectivity. The following section deals with experimental results of URFL cavities. The enhancement of the available spectral bandwidth in the C-band and its spectral flatness are investigated for single and multi-FBGs cavity system. Further work regarding extended URFL cavity in combination with Rayleigh scattering as random distributed feedback produced a laser cavity with dual wavelength outputs independent to each other. The last two sections relate to URFL application in supercontinuum (SC) generation and soliton transmission. URFL becomes an enhancement structure for SC generation. This thesis shows successful experimental results of SC generation using conventional single mode optical fibre and pumped with a continuous wave source. The last section is dedicated to soliton transmission and the study of soliton propagation dynamics. The experimental results of exact soliton transmission over multiple soliton periods using conventional single mode fibre are shown in this thesis. The effect of the input signal, pump distribution, span length and FBGs reflectivity on the soliton propagation dynamics is investigated experimentally and numerically.

Clinical studies of spatial and temporal aspects of vision: an investigation using psychophysical and electrophysiological techniques

Separate physiological mechanisms which respond to spatial and temporal stimulation have been identified in the visual system. Some pathological conditions may selectively affect these mechanisms, offering a unique opportunity to investigate how psychophysical and electrophysiological tests reflect these visual processes, and thus enhance the use of the tests in clinical diagnosis. Amblyopia and optical blur were studied, representing spatial visual defects of neural and optical origin, respectively. Selective defects of the visual pathways were also studied - optic neuritis which affects the optic nerve, and dementia of the Alzheimer type in which the higher association areas are believed to be affected, but the primary projections spared. Seventy control subjects from 10 to 79 years of age were investigated. This provided material for an additional study of the effect of age on the psychophysical and electrophysiological responses. Spatial processing was measured by visual acuity, the contrast sensitivity function, or spatial modulation transfer function (MTF), and the pattern reversal and pattern onset-offset visual evoked potential (VEP). Temporal, or luminance, processing was measured by the de Lange curve, or temporal MTF, and the flash VEP. The pattern VEP was shown to reflect the integrity of the optic nerve, geniculo striate pathway and primary projections, and was related to high temporal frequency processing. The individual components of the flash VEP differed in their characteristics. The results suggested that the P2 component reflects the function of the higher association areas and is related to low temporal frequency processing, while the Pl component reflects the primary projection areas. The combination of a delayed flash P2 component and a normal latency pattern VEP appears to be specific to dementia of the Alzheimer type and represents an important diagnostic test for this condition.