Bases moleculares de la resistencia a insecticidas en la mosca mediterránea de la fruta Ceratitis Capitata (Wiedemann)

La mosca mediterránea de la fruta Ceratitis capitata (Wiedemann, 1824) está considerada una de las plagas clave para la fruticultura. El malatión es un insecticida organofosforado que fue empleado mayoritariamente en España para el control de C. capitata hasta 2009, año en el que dejó de utilizarse por no estar incluido en el anexo I de la Directiva Europea 91/414/ECC. El incremento del uso del malatión, debido a las graves pérdidas económicas causadas por C. capitata, provocó la aparición de poblaciones de campo resistentes. El estudio de una población resistente a malatión, recogida en Castelló en 2004, permitió la identificación de dos mecanismos de resistencia: una mutación puntual (G328A) en la acetilcolinesterasa (AChE) y un mecanismo de resistencia metabólica, probablemente mediado por carboxilesterasas. Teniendo en cuenta estos antecedentes, nos propusimos estudiar los mecanismos implicados en la resistencia a malatión en C. capitata. Además, durante el desarrollo de esta Tesis, el malatión fue sustituido por otros insecticidas como el espinosad y la lambda-cialotrina para el control de la plaga. En este nuevo contexto, es extremadamente importante analizar la susceptibilidad de poblaciones de campo frente a espinosad y estudiar la posible existencia de resistencia cruzada a estos insecticidas, así como sentar las bases para el estudio de futuros mecanismos de resistencia. En primer lugar, analizamos mediante bioensayos con dosis discriminante la susceptibilidad a malatión y espinosad en doce poblaciones de C. capitata de Andalucía, Aragón, Cataluña, Comunidad Valenciana e Islas Baleares; y nuestros resultados sugirieron la presencia de individuos resistentes a malatión en la mayoría de las poblaciones analizadas. En el caso del espinosad, observamos que la susceptibilidad a este insecticida de origen biológico fue elevada en la mayoría de las poblaciones, sin embargo, la población recogida en Xàbia (Alicante) mostró un nivel de susceptibilidad unas dos veces menor al resto de poblaciones. Mediante la selección en laboratorio, obtuvimos dos líneas resistentes a malatión, W-4Km y W-10Km, con unos niveles de resistencia con respeto a la línea susceptible C de 178 y 400 veces, respectivamente. Además, se seleccionó por primera vez en C. capitata una línea altamente resistente a espinosad (Xàbia-W-100s), que actualmente es unas 500 veces más resistente que la línea de laboratorio C. Con el objetivo de escoger la estrategia más adecuada para el manejo de la plaga, estudiamos la susceptibilidad a diferentes tipos de insecticidas en la línea resistente a malatión W- 4Km. En esta línea detectamos resistencia cruzada moderada a los organofosforados fentión, diazinón, fosmet, triclorfón y metil-clorpirifos (de 7 a 16 veces) y frente al carbamato carbaril, al piretroide lambda-cialotrina y al quimioesterilizante lufenurón (de 4 a 6 veces). Por otra parte, la resistencia cruzada frente a espinosad fue baja (1,5 veces). Es importante destacar que los niveles de resistencia estimados frente a todos los insecticidas fueron de uno o dos órdenes de magnitud inferiores al observado en la línea W-4Km frente a malatión (178 veces), hecho que podría deberse, al menos, a dos posibles hipótesis: que la mutación AChE G328A confiera mayor insensibilidad al malaoxón (forma activa del malatión) que a otros insecticidas que tienen como diana la AChE y/o, en segundo lugar, que el mecanismo de resistencia mediado por carboxilesterasas hidrolice el malatión de manera más eficiente que los otros insecticidas analizados. En el estudio de nuevos mecanismos de resistencia en C. capitata, por un lado, analizamos la diversidad de enzimas citocromo P450, asociadas con resistencia metabólica en otras especies, y por otro lado, desarrollamos un sistema para la detección de nuevas mutaciones puntuales que pudiesen aparecer en los genes que codifican la AChE (Ccace2) y la aliesterasa (Ccae7). Mediante el empleo de cebadores degenerados obtuvimos 37 genes CYP, que codifican enzimas P450, pertenecientes a cinco familias. Posteriormente, en un estudio de inducción con fenobarbital, observamos que la expresión de cuatro de los seis genes analizados era susceptible de ser inducida. Por otro lado, se puso a punto un sistema que permite amplificar y secuenciar, a partir de DNA genómico, los exones de los genes Ccace2 y Ccae7 en los que se han encontrado mutaciones relacionadas con resistencia a insecticidas en otras especies. Los resultados obtenidos facilitarán el estudio de nuevos mecanismos de resistencia mediados por estas enzimas en C. capitata. Se diseñó un método PCR-RFLP para identificar los individuos portadores de la mutación AChE G328A (alelo de resistencia Ccace2R) sin la necesidad de realizar bioensayos y que, además, permite detectar resistencia cuando ésta se encuentra a baja frecuencia. Según el análisis realizado, el alelo Ccace2R se observó en 25 de las 27 localidades españolas muestreadas en el territorio español, incluyendo las Islas Baleares y Canarias. Sin embargo, este alelo no se detectó en poblaciones procedentes de once países y de cinco continentes. El análisis de la presencia del alelo Ccace2R en las líneas resistentes a malatión durante el proceso de selección en el laboratorio mostró una rápida disminución de los homocigotos, tanto para el alelo susceptible como para el alelo de resistencia, en favor de los individuos heterocigotos. Así, después de 52 generaciones de selección, se observó que la totalidad de los individuos analizados de la línea W-10Km presentaban un genotipo heterocigoto para la mutación AChE G328A. Este desequilibrio contradice la segregación mendeliana esperada para un gen con dos alelos pero podría ser explicado por la existencia de una duplicación del gen Ccace2. La demostración de la presencia de esta duplicación se realizó mediante: i) el cruzamiento de individuos heterocigotos de la línea W-10Km con homocigotos susceptibles de la línea C, que dio lugar a una descendencia en la que el 100% de los individuos eran heterocigotos; ii) la evaluación del número de copias del gen Ccace2 por PCR cuantitativa en tiempo real (qPCR), que resultó dos veces mayor en individuos de la línea W-10Km en comparación con los de la línea C; iii) el análisis del nivel de expresión de Ccace2, que fue el doble en la línea W-10Km con respecto a la línea C, y iv) el estudio de la actividad AChE, que resultó mayor en los individuos de la línea W-10Km. Según los resultados obtenidos, una duplicación del gen Ccace2 provoca la coexistencia en un mismo cromosoma del alelo silvestre y del alelo mutado y, además, las dos copias del gen Ccace2, al estar ligadas, producen una heterocigosis permanente (Ccace2RS). De esta manera se explica que el hecho de que 100% de los individuos de la línea W-10Km mostrasen un perfil de restricción correspondiente a un individuo heterocigoto ya que, en realidad, eran homocigotos estructurales para la duplicación (genotipo CCace2RS/RS). Se ha detectado un coste biológico asociado a la duplicación que consiste en un incremento en la mortalidad acumulada de los adultos a partir del séptimo día después de la emergencia. La descripción de la duplicación Ccace2RS supone la identificación de un nuevo mecanismo de resistencia a malatión en C. capitata. Finalmente, mediante el diseño de un método de doble PCR-RFLP se determinó la presencia de la duplicación Ccace2RS en la mayoría de las poblaciones españolas. La proporción de individuos portadores de la duplicación osciló entre el 5% y el 35%, observándose los mayores valores de frecuencia en las poblaciones de C. capitata recogidas en la cuenca mediterránea. Podemos por lo tanto concluir que la resistencia a malatión asociada a la mutación AChE G328A y a la duplicación Ccace2RS está ampliamente establecida en las poblaciones españolas de C. capitata. Nuestros resultados desaconsejan la utilización del malatión (si fuera de nuevo autorizado) o de otros organofosforados para el control de esta plaga. Además, una de las líneas resistentes a malatión mostró resistencia cruzada frente a insecticidas con diferentes modos de acción y que se utilizan actualmente para el control de C. capitata, tales como lambda-cialotrina y lufenurón. La alta susceptibilidad a espinosad observada en las poblaciones españolas, así como la reducida resistencia cruzada estimada para este insecticida, sugieren que su utilización es adecuada para el control de la plaga. Sin embargo, la utilización de un sólo insecticida puede entrañar riesgos por favorecer la selección de resistencia, de hecho, mediante selección en laboratorio se obtuvo una población altamente resistente a espinosad. Por tanto, es recomendable implementar programas de control integrado y de manejo de la resistencia en C. capitata utilizando distintos sistemas de control e insecticidas con diferentes mecanismos de acción que permitan su sostenibilidad en el tiempo. Los sistemas de detección de alelos de resistencia desarrollados en este trabajo permitirán la detección precoz de resistencia en campo, facilitando la decisión sobre el sistema de control más adecuado. Además, los conocimientos generados podrán contribuir al desarrollo de nuevos sistemas de detección para otros mecanismos de resistencia. Abstract. The Mediterranean fruit fly, Ceratitis capitata (Wiedemann, 1824), is considered one of the most harmful pests in fruit crops. Until 2009, when malathion use was banned due to its not inclusion in the Annex I of Directive 91/414/EEC, the application of this organophosphate (OP) insecticide in Spain increased gradually due to the large economic losses caused by C. capitata. The increase in the frequency of treatments resulted in the development of resistant field populations. The study of a malathion-resistant population, collected in 2004 in Castelló (Comunidad Valenciana), allowed the identification of two resistance mechanisms: a single point mutation (G328A) in the target acetylcholinesterase (AChE), as well as a metabolic resistance mechanism, most likely carboxylesterase-mediated. Taking all the preceding into account, we studied the malathion resistance mechanisms in C. capitata. During the development of this PhD Thesis malathion use was banned by the European Union, being replaced by other insecticides, such as spinosad and lambda-cyhalotrin. Within this new working frame, the need to analyse the possible existence of cross-resistance to these insecticides and the susceptibility to spinosad in field populations was raised. This would define the baseline for future studies on resistance mechanisms. Firstly, through discriminant dose bioassays, we analysed malathion and spinosad susceptibility in twelve C. capitata populations from Andalucia, Aragon, Cataluña, C. Valenciana and the Baleares Islands. Our results suggest the presence of malathion-resistant individuals in most of the populations analysed. Regarding spinosad, we noticed a high susceptibility to this biologically derived insecticide in most of the populations, but in the one collected in Xabia (Alicante), which had a susceptibility level two times lower than the rest of populations. Through laboratory selection, we obtained two malathion-resistant strains, W-4Km and W-10Km, with resistance levels 178- and 400-fold, respectively, compared to the control susceptible C strain. Besides, a strain highly-resistant to spinosad (Xabia-W-100s), 500-times more resistant than control C strain, was selected. In order to decide the most appropriate management strategy for the pest, we studied the susceptibility to different insecticides in the malathion-resistant W-4Km strain. We detected a moderated cross-resistance to the OPs fenthion, diazinon, phosmet, trichlorphon and methylchlorpyrifos (7- to 16-fold), and to the carbamate carbaryl, the pyretroid lambda-cyhalotrin and the chemosterilizer lufenuron (4- to 6-fold). On the other hand, cross-resistance to spinosad was low (1.5-fold). It is important to note that resistance levels to all insecticides were one or two orders of magnitude less than that observed against malathion in W-4Km strain (178-fold), a fact that might be due to, at least, two possible causes: mutation AChE G328A may provide a higher insensitivity to malaoxon (the active form of malathion) than to other insecticides having AChE as target, and/or, secondly, the carboxylesterase-mediated resistance mechanism hydrolyzes malathion more efficiently than all other analysed insecticides. To investigate new resistance mechanisms in C. capitata we analysed the diversity of the cytochrome P450 enzymes, which have been associated to metabolic resistance in insects, and we developed a new method to detect single point mutations in acetylcholinesterase (Ccace2) and aliesterase (Ccae7) genes that could appear. Using degenerate primers we obtained 37 CYP genes, coding P450 enzymes, included in five families. Afterwards, in a phenobarbital-induction study, we observed that the expression of 4 out of the 6 analysed genes could be induced. On the other hand, a system was set up to amplify and to sequence from genomic DNA the exons of genes Ccace2 and Ccae7 where mutations related to insecticide resistance have been found in other species. The results obtained could facilitate the study of new resistance mechanisms in C. capitata mediated by these enzymes. A PCR-RFLP method was designed to detect the presence of the mutation AChE G328A (resistance allele Ccace2R), with no need to perform bioassays and allowing detecting resistance at low frequency. According to the analysis, the resistance allele was found in 25 out of 27 sampled locations in Spain, including the Balearic and the Canary Islands. However, this allele was not detected in other populations collected in 11 countries from 5 continents. The follow-up of the presence of the allele Ccace2R in the malathion-resistant strains during the selection process in the laboratory showed a quick decrease in homozygous individuals, for both the susceptible and the resistant alleles, favouring heterozygous. Thus, after 52 generations of selection, all the individuals analysed from W-10Km strain showed a heterozygous genotype for mutation AChE G328A, contradicting mendelian segregation as expected for a gene with two alleles. Afterwards, we were able to demonstrate that this was caused by the presence of a duplication of the gene coding acetylcholinesterase by: i) crossing heterozygous individuals from W-10Km strain with susceptible homozygous from C strain, originating a F1 population in which 100% of individuals were heterozygous; ii) evaluating the number of copies of gen Ccace2 by quantitative PCR in real time (qPCR), that happened to be twice higher in individuals from W-10Km VII strain when compared with C strain; iii) analysing the level of expression of Ccace2, twice in W- 10Km strain when compared to C strain; iv) studying the acetylcholinesterase activity, that was higher in individuals from W-10Km strain. According to these results, duplication of gen Ccace2 originates the coexistence of the susceptible and the resistant allele in the same chromosome. The two linked copies of the gene Ccace2 provoke the existence of permanent heterozygosis (Ccace2RS). This explains why the 100% of individuals from W-10Km strain showed an heterozygous restriction pattern since, in fact, they were structural homozygotes for the duplication (genotype Ccace2RS/RS). A biological cost has been detected associated to this duplication, consisting in a rise in accumulated adult mortality from the seventh day after emergence. The Ccace2RS duplication described in this study represents a new resistance mechanism to malathion in C. capitata. Finally, by the design of a double PCR-RFLP method, the presence of Ccace2RS duplication was confirmed in most of the Spanish populations. We observed that the proportion of individuals carrying the duplication oscillated between 5 and 35%, the frequency being higher in those C. capitata populations collected in the area of the Mediterranean basin. Therefore, we can conclude that malathion resistance associated to mutation AChE G328A and to Ccace2RS duplication are widely distributed in Spanish populations of C. capitata. Our results advice against the use of malathion (if it came to be newly authorized for use) or other OPs for the control of this pest. Besides, one of the malathion-resistant strains showed cross-resistance against insecticides with diverse action modes that are currently used for pest control, such as lambdacyhalotrin and lufenuron. High susceptibility to spinosad in the Spanish populations, as well as the reduced cross-resistance estimated for this insecticide suggests its adequacy for Medfly control. However, the use of a single insecticide is a risky strategy since it favours the selection of resistance. In fact, a population highly resistant to spinosad was obtained through laboratory selection. Therefore, it is advisable to implement integrated pest management (IPM) and resistance management programs for C. capitata control. Using insecticides with different modes of action and diverse control systems would contribute to the sustainability of the pest control. The resistance allele detection systems developed through this work will allow the early detection of resistance in the field, making possible the selection of the most appropriate method for pest control. Besides, the generated knowledge may also contribute to the development of new detection systems for other resistance mechanisms.

Hysteresis phenomena in transverse galloping of triangular cross-section bodies

Transverse galloping is a type of aeroelastic instability characterised by large amplitude, low frequency oscillation of a structure in the direction normal to the mean wind direction. It normally appears in bodies with small stiffness and structural damping, provided the incident flow velocity is high enough. In the simplest approach transverse galloping can be considered as a one-degree-of-freedom oscillator subjected to aerodynamic forces, which in turn can be described by using a quasi-steady description. In this frame it has been demonstrated that hysteresis phenomena in transverse galloping is related to the existence of inflection points in the curve giving the dependence with the angle of attack of the aerodynamic coefficient normal to the incident flow. Aiming at experimentally checking such a relationship between these inflection points and hysteresis, wind tunnel experiments have been conducted. Experiments have been restricted to isosceles triangular cross-section bodies, whose galloping behaviour is well documented. Experimental results show that, according to theoretical predictions, hysteresis takes place at the angles of attack where there are inflection points in the lift coefficient curve, provided that the body is prone to gallop at these angles of attack.

Rheological and tribological properties of poly(phenylene sulphide) and poly(ether ether ketone) composites incorporating carbon nanotubes and inorganic nanoparticles: a comparative study

The rheological and tribological properties of single-walled carbon nanotube (SWCNT)-reinforced poly(phenylene sulphide) (PPS) and poly(ether ether ketone) (PEEK) nanocomposites prepared via melt-extrusion were investigated. The effectiveness of employing a dual-nanofiller strategy combining polyetherimide (PEI)-wrapped SWCNTs with inorganic fullerene-like tungsten disulfide (IF-WS2) nanoparticles for property enhancement of the resulting hybrid composites was evaluated. Viscoelastic measurements revealed that the complex viscosity ?, storage modulus G?, and loss modulus G? increased with SWCNT content. In the low-frequency region, G? and G? became almost independent of frequency at higher SWCNT loadings, suggesting a transition from liquid-like to solid-like behavior. The incorporation of increasing IF-WS2 contents led to a progressive drop in ? and G? due to a lubricant effect. PEEK nanocomposites showed lower percolation threshold than those based on PPS, ascribed to an improved SWCNT dispersion due to the higher affinity between PEI and PEEK. The SWCNTs significantly lowered the wear rate but only slightly reduced the coefficient of friction. Composites with both nanofillers exhibited improved wear behavior, attributed to the outstanding tribological properties of these nanoparticles and a synergistic reinforcement effect. The combination of SWCNTs with IF-WS2 is a promising route for improving the tribological and rheological performance of thermoplastic nanocomposites.

Novel system for bite-force sensing and monitoring based on magnetic eear field communication

Intraoral devices for bite-force sensing have several applications in odontology and maxillofacial surgery, as bite-force measurements provide additional information to help understand the characteristics of bruxism disorders and can also be of help for the evaluation of post-surgical evolution and for comparison of alternative treatments. A new system for measuring human bite forces is proposed in this work. This system has future applications for the monitoring of bruxism events and as a complement for its conventional diagnosis. Bruxism is a pathology consisting of grinding or tight clenching of the upper and lower teeth, which leads to several problems such as lesions to the teeth, headaches, orofacial pain and important disorders of the temporomandibular joint. The prototype uses a magnetic field communication scheme similar to low-frequency radio frequency identification (RFID) technology (NFC). The reader generates a low-frequency magnetic field that is used as the information carrier and powers the sensor. The system is notable because it uses an intra-mouth passive sensor and an external interrogator, which remotely records and processes information regarding a patient?s dental activity. This permits a quantitative assessment of bite-force, without requiring intra-mouth batteries, and can provide supplementary information to polysomnographic recordings, current most adequate early diagnostic method, so as to initiate corrective actions before irreversible dental wear appears. In addition to describing the system?s operational principles and the manufacture of personalized prototypes, this report will also demonstrate the feasibility of the system and results from the first in vitro and in vivo trials.

Towards the development of poly (phenylene sulphide) based nanocomposites with enhanced mechanical, electrical and tribological properties

Novel poly(phenylene sulphide) (PPS) nanocomposites reinforced with an aminated derivative (PPS-NH2) covalently attached to acid-treated single-walled carbon nanotubes (SWCNTs) were prepared via simple melt-blending technique. Their morphology, viscoelastic behaviour, electrical conductivity, mechanical and tribological properties were investigated. Scanning electron microscopy revealed that the grafting process was effective in uniformly dispersing the SWCNTs within the matrix. The storage and loss moduli as a function of frequency increased with the SWCNT content, tending to a plateau in the low-frequency regime. The electrical conductivity of the nanocomposites was considerably enhanced in the range 0.1?0.5 wt% SWCNTs; electrical and rheological percolation thresholds occurred at similar nanotube concentrations. Mechanical tests demonstrated that with only 1.0 wt% SWCNTs the Young's modulus and tensile strength of the matrix improved by 51 and 37%, respectively, without decrement in toughness, ascribed to a very efficient load transfer. A moderate decrease in the friction coefficient and a 75% reduction in wear rate were found for the abovementioned nanotube loading, indicating that PPS-NH2-g-SWCNTs are good tribological additives for thermoplastic polymers. Based on the promising results obtained in this work, it is expected that these nanofillers will be used to develop high-performance thermoplastic/CNT nanocomposites for structural applications.

Synchronous Buck converter with Output Impedance Correction Circuit

This work is related to the improvement of the output impedance of the Buck converter by means of introducing an additional power path that virtually increases the output capacitance during transients. It is well known that in VRM applications, with wide load steps, voltage overshoots and undershoots may lead to undesired performance of the load. To solve this problem, high-bandwidth high-switching frequency power converters can be applied to reduce the transient time or a big output capacitor can be applied to reduce the output impedance. The first solution can degrade the efficiency by increasing switching losses of the MOSFETS, and the second solution is penalizing the cost and size of the output filter. The Output Impedance Correction Circuit (OICC), as presented here, is used to inject or extract a current n-1 times larger than the output capacitor current, thus virtually increasing n times the value of the output capacitance during the transients. This feature allows the usage of a low frequency Buck converter with smaller capacitor but satisfying the dynamic requirements.

Improvement of the Dynamic Performance of a Buck Converter using Controlled Current Source as an Additional Energy Path

This work is related to the improvement of the dynamic performance of the Buck converter by means of introducing an additional power path that virtually increase s the output capacitance during transients, thus improving the output impedance of the converter. It is well known that in VRM applications, with wide load steps, voltage overshoots and undershoots ma y lead to undesired performance of the load. To solve this problem, high-bandwidth high-switching frequency power converter s can be applied to reduce the transient time or a big output capacitor can be applied to reduce the output impedance. The first solution can degrade the efficiency by increasing switching losses of the MOSFETS, and the second solution is penalizing the cost and size of the output filter. The additional energy path, as presented here, is introduced with the Output Impedance Correction Circuit (OICC) based on the Controlled Current Source (CCS). The OICC is using CCS to inject or extract a current n - 1 times larger than the output capacitor current, thus virtually increasing n times the value of the output capacitance during the transients. This feature allows the usage of a low frequency Buck converter with smaller capacitor but satisfying the dynamic requirements.

Análisis del acoplamiento vibro-acústico en resonadores de membrana

El presente Trabajo Fin de Máster pretende llevar a cabo el análisis del comportamiento vibratorio de resonadores de membrana, consistentes en un panel delgado y ligero montado a cierta distancia de un elemento constructivo rígido y pesado. Este tipo de sistemas resonantes son empleados habitualmente como absorbentes de media-baja frecuencia en aplicaciones de acondicionamiento acústico de salas. El análisis hará especial hincapié en la influencia del acoplamiento mecánico-acústico entre la placa vibrante (estructura) y el colchón de aire (fluido) encerrado entre la misma y la pared rígida. En primer lugar, realizaremos el análisis modal experimental del resonador objeto de ensayo a partir de las mediciones de su respuesta vibratoria, con el fin de caracterizar su comportamiento en base a sus primeros modos propios acoplados de flexión. El análisis de las señales vibratorias en el dominio de la frecuencia para la identificación de dicho modos se realizará en el entorno de programación MATLAB, haciendo uso de una herramienta propia que implementa los métodos de cálculo y los algoritmos necesarios para tal fin. Asimismo, simularemos el comportamiento del resonador mediante el método de elementos finitos (FEM), utilizando las aplicaciones ANSYS y SYSNOISE, considerando diferentes condiciones frontera en el modelo generado. Los resultados aquí obtenidos serán de utilidad para complementar aquellos obtenidos de forma experimental a la hora de extraer conclusiones prácticas del análisis realizado. SUMMARY. This Master's Thesis intends to carry out the analysis of the vibratory behaviour of resonance absorbers, consisting of a thin and lightweight panel mounted at a distance from a rigid wall. Such systems are commonly used as sound absorption systems for mid-low frequency in room acoustics applications. The analysis will emphasize the influence of mechanical-acoustic coupling between the vibrating plate (structure) and the air cushion (acoustic element) enclosed behind it. First of all, we are performing the experimental modal analysis of the resonance absorber under test from the vibrational response measurements, in order to characterize its behaviour based on its first bending coupled-modes. The analysis of vibration signals in the frequency domain for the identification of such modes will be made in MATLAB programming environment, using a proprietary tool that implements the calculation methods and algorithms needed for this purpose. Furthermore, we are simulating the behaviour of the resonance absorber applying the Finite Element Method (FEM) – using ANSYS and SYSNOISE applications - considering different boundary conditions in the model created. The results from the simulation will be useful to complement those obtained experimentally when drawing practical conclusions from this analysis.

Contribución al estudio de sistemas de hipertermia óptica y su aplicación en muestras biológicas

Las nanopartículas de metales nobles (especialmente las de oro) tienen un gran potencial asociado al desarrollo de sistemas de terapia contra el cáncer debido principalmente a sus propiedades ópticas, ya que cuando son irradiadas con un haz de luz sintonizado en longitud de onda con su máximo de Resonancia de Plasmón Superficial, absorben de manera muy eficiente dicha luz y la disipan rápidamente al medio en forma de calor localizado. Esta característica por tanto, puede ser aprovechada para conseguir elevar la temperatura de células tumorales hasta sobrepasar umbrales a partir de los cuales se produciría la muerte celular. Partiendo de estos principios, esta tesis se centra en el desarrollo y la caracterización de una serie de prototipos de hipertermia óptica basados en la irradiación de nanopartículas de oro con un haz de luz adecuado, así como en la aplicación in vitro de la terapia sobre células cancerígenas. Además, el trabajo se orienta a identificar y comprender los procesos mecánicos y térmicos asociados a este tipo de hipertermia, y a desarrollar modelos que los describan, estudiando y planteando nuevas formas de irradiación, para, en última instancia, poder optimizar los procesos descritos y hacerlos más efectivos. Los resultados obtenidos indican que, el uso de nanopartículas de oro, y más concretamente de nanorods de oro, para llevar a cabo terapias de hipertermia óptica, permite desarrollar terapias muy efectivas para inducir muerte en células cancerígenas, especialmente en tumores superficiales, o como complemento quirúrgico en tumores internos. Sin embargo, los efectos de la toxicidad de las nanopartículas de oro, aún deben ser detalladamente estudiados, ya que este tipo de terapias sólo será viable si se consigue una completa biocompatibilidad. Por otro lado, el estudio exhaustivo de los procesos térmicos que tienen lugar durante la irradiación de las nanopartículas ha dado lugar a una serie de modelos que permiten determinar la efectividad fototérmica de las nanopartículas y además, visualizar la evolución de la temperatura tanto a escala nanométrica como a escala macrométrica, en función de los parámetros ópticos y térmicos del sistema. El planteamiento de nuevas formas de irradiación y el desarrollo de dispositivos orientados a estudiar los fenómenos mecánicos que tienen lugar durante la irradiación pulsada de baja frecuencia y baja potencia de nanopartículas de oro, ha dado lugar a la detección de ondas de presión asociadas a procesos de expansión termoelástica, abriendo la puerta al desarrollo de terapias de hipertermia que combinen la muerte celular producida por calentamiento con la muerte derivada de los fenómenos mecánicos descritos.VII Noble metal nanoparticles (especially gold ones), have a huge potential in the development of therapy systems against cancer mainly due to their optical properties, so that, when these particles are irradiated with a light that is syntonized in wavelength with their maximum of Surface Plasmon Resonance, they effectively absorb and dissipate the light to the surrounding medium as localized heat. We can take advantage of this characteristic for rising the temperature of cancer cells above the threshold at which cellular death would occur. From these principles, this thesis is oriented to the development and characterization of a series of optical hyperthermia prototypes based on the irradiation of gold nanoparticles using the suitable light, and on the in vitro application of this therapy over cancer cells, to understand the mechanical and thermal processes associated with this kind of hyperthermia, developing descriptive models, and to study and to approach new ways of irradiation in order to, ultimately, optimize the described processes and make them more effective. The obtained results show that, the use of gold nanoparticles, and more specifically, of gold nanorods, to carry out optical hyperthermia therapies, allows the development of very effective therapies in order to induce death in VIII cancer cells, especially in superficial tumors, or like surgical complement in more internal tumors. However, the toxicity effects of the gold nanoparticles still need to be studied more detail, because this kind of therapies will be feasible only if a complete biocompatibility is achieved. On the other hand, the exhaustive study of the thermal processes that take place during the irradiation of the nanoparticles resulted in a series of models that allow the determination of the photothermal efficiency of the nanoparticles and also the visualization of the temperature evolution, both at nanoscale and at macroscale, as a function of the optical and thermal parameters of the system. The proposal of new ways of irradiation and the development of devices oriented to study the mechanical effects that take place during the low frequency and low power pulsing irradiation of gold nanoparticles has led to the detection of pressure waves associated to thermoelastic expansion processes, opening the door to the development of hyperthermia therapies that combine the cellular death due to the heating with the death derived from the described mechanical phenomena.

Electrohydrodynamic behavior in twisted-wedge nematic structures

A new type of domain for nematic liquid crystals with a twisted-wedge structure is presented. This new type of domain appears from the low frequency range to 10 kHz. This behavior was observed for square and pulsed excitations. The liquid crystal was N-(p-methoxybenzylidene)-p'-butylaniline) (MBBA) used at room temperature. These domains offer a higher degree of complexity than conventional Williams domains. The corresponding stability chart is presented.

Fast magnetosonic wave excitation by an array of wires with time-modulated currents

The excitation of Fast Magnetosonic (FMS)waves by a cylindrical array of parallel tethers carrying timemodulated current is discussed. The tethers would fly vertical in the equatorial plane, which is perpendicular to the geomagnetic field when its tilt is ignored, and would be stabilized by the gravity gradient. The tether array would radiate a single FMS wave. In the time-dependent background made of geomagnetic field plus radiated wave, plasma FMS perturbations are excited in the array vicinity through a parametric instability. The growth rate is estimated by truncating the evolution equation for FMS perturbations to the two azimuthal modes of lowest order. Design parameters such as tether length and number, required power and mass are discussed for Low Earth Orbit conditions. The array-attached wave structure would have the radiated wave controlled by the intensity and modulation frequency of the currents, making an active experiment on non-linear low frequency waves possible in real space plasma conditions.

Estudio de la vibración en baja frecuencia de motor eléctrico

Se realizará un análisis en baja frecuencia del comportamiento de sistemas vibratorios excitados principalmente por un motor eléctrico. El sistema está formado por un motor eléctrico acoplado a un volante de inercia que produce una carga sobre citado motor diferente según las configuraciones adoptadas, todo ello montado sobre una base metálica de acero, soportada esta, mediante resortes de diferente características. El estudio contemplará la identificación de frecuencias de excitación, resonancias, pérdidas de inserción de los sistemas, transmisibilidad, problemas de alineamiento, desajustes, modos propios... para cada una de las diferentes situaciones en las que opera el sistema. ABSTRACT. In this Project will proceed to an behavior analysis of vibrating systems in low frequency mainly excited by an electric motor. The system is comprised of an electric motor coupled to inertial flywheel (o flywheel) which produces a different load upon said engine according to the configurations adopted. This system is mounted on a steel metal base which is supported by springs of different characteristics. This study will consider the excitation frequency identification, system resonances, insertion loss, transmissibility, shaft dealignment, eccentricities, mismatches, modal frequencies of the plate… for each of the situations in which the system operates.

Using array based techniques for noise source identification in cars

Una de las principales causas del ruido en nuestras ciudades es el tráfico rodado. El ruido generado por los vehículos no es sólo debido al motor, sino que existen diversas fuentes de ruido en los mismos, entre las que se puede destacar el ruido de rodadura. Para localizar las causas del ruido e identificar las principales fuentes del mismo se han utilizado en diversos estudios las técnicas de coherencia y las técnicas basadas en arrays. Sin embargo, en la bibliografía existente, no es habitual encontrar el uso de estas técnicas en el sector automovilístico. En esta tesis se parte de la premisa de la posibilidad de usar estas técnicas de medida en coches, para demostrar a la largo de la misma su factibilidad y su bondad para evaluar las fuentes de ruido en dos condiciones distintas: cuando el coche está parado y cuando está en movimiento. Como técnica de coherencia se elige la de Intensidad Selectiva, utilizándose la misma para evaluar la coherencia existente entre el ruido que llega a los oídos del conductor y la intensidad radiada por distintos puntos del motor. Para la localización de fuentes de ruido, las técnicas basadas en array son las que mejores resultados ofrecen. Statistically Optimized Near-field Acoustical Holography (SONAH) es la técnica elegida para la localización y caracterización de las fuentes de ruido en el motor a baja frecuencia. En cambio, Beamforming es la técnica seleccionada para el caso de media-alta frecuencia y para la evaluación de las fuentes de ruido cuando el coche se encuentra en movimiento. Las técnicas propuestas no sólo pueden utilizarse en medidas reales, sino que además proporcionan abundante información y frecen una gran versatilidad a la hora de caracterizar fuentes de ruido. ABSTRACT One of the most important noise causes in our cities is the traffic. The noise generated by the vehicles is not only due to the engine, but there are some other noise sources. Among them the tyre/road noise can be highlighted. Coherence and array based techniques have been used in some research to locate the noise causes and identify the main noise sources. Nevertheless, it is not usual in the literature to find the application of this kind of techniques in the car sector. This Thesis starts taking into account the possibility of using this kind of measurement techniques in cars, to demonstrate their feasability and their quality to evaluate the noise sources under two different conditions: when the car is stopped and when it is in movement. Selective Intensity was chosen as coherence technique, evaluating the coherence between the noise in the driver’s ears and the intensity radiated in different points of the engine. Array based techniques carry out the best results to noise source location. Statistically Optimized Near-field Acoustical Holography (SONAH) is the measurement technique chosen for noise source location and characterization in the engine at low frequency. On the other hand, Beamforming is the technique chosen in the case of medium-high frequency and to characterize the noise sources when the car is in movement. The proposed techniques not only can be used in actual measurements, but also provide a lot of information and are very versatile to noise source characterization.

Pulsating emission of droplets from an electrified meniscus.

A numerical description is given for the pulsating emission of droplets from an electrified meniscus of an inviscid liquid of infinite electrical conductivity which is injected at a constant flow rate into a region of uniform, continuous or time periodic, electric field. Under a continuous field, the meniscus attains a periodic regime in which bursts of tiny droplets are emitted from its tip. At low electric fields this regime consists of sequences of emission bursts interspersed with sequences of meniscus oscillations without droplet emission, while at higher fields the bursts occur periodically. These results are in qualitative agreement with experimental results in the literature. Under a time periodic electric field with square waveform, the electric stress that acts on the surface of the liquid while the field is on may generate a tip that emits tiny droplets or may accelerate part of the meniscus and lead to a second emission mode in which a few large droplets are emitted after the electric field is turned off. Conditions under which each emission mode or a combination of the two are realized are discussed for low frequency oscillatory fields. A simplified model is proposed for high electric field frequencies, of the order of the capillary frequency of the meniscus. This model allows computing the average emission rate as a function of the amplitude, duration and bias of the electric field square wave, and shows that droplet emission fails to follow the applied field above a certain frequency

InfIuence of wave induced second-order forces in semisubmersible fowt mooring design

The main focus of this paper is on hydrodynamic modelling of a semisubmersible platform (which can support a 1.5MW wind turbine and is composed by three buoyant columns connected by bracings) with especial emphasis on the estimation of the wave drift components and their effects on the design of the mooring system. Indeed, with natural periods of drift around 60 seconds, accurate computation of the low-frequency second-order components is not a straightforward task. As methods usually adopted when dealing with the slow-drifts of deep-water moored systems, such as Newman?s approximation, have their errors increased by the relatively low resonant periods, and as the effects of depth cannot be ignored, the wave diffraction analysis must be based on full Quadratic Transfer Functions (QTF) computations. A discussion on the numerical aspects of performing such computations is presented, making use of the second-order module available with the seakeeping software WAMIT®. Finally, the paper also provides a preliminary verification of the accuracy of the numerical predictions based on the results obtained in a series of model tests with the structure fixed in bichromatic waves.