Determination of drugs used in combined cardiovascular therapy

Cardiovascular diseases are nowadays the first cause of mortality worldwide, causing around the 30% of global deaths each year. The risk of suffering from cardiovascular illnesses is strongly related to some factors such as hypertension, high cholesterol levels, diabetes, obesity The combination of these different risk factors is known as metabolic syndrome and it is considered a pandemic due to the high prevalence worldwide. The pathology of the disorders implies a combined cardiovascular therapy with drugs which have different targets and mechanisms of action, to regulate each factor separately. The simultaneous analysis of these drugs turns interesting but it is a complex task since the determination of multiple substances with different physicochemical properties and physiological behavior is always a challenge for the analytical chemist. The complexity of the biological matrices and the difference in the expected concentrations of some analytes require the development of extremely sensitive and selective determination methods. The aim of this work is to fill the gap existing in this field of the drug analysis, developing analytical methods capable of quantifying the different drugs prescribed in combined cardiovascular therapy simultaneously. Liquid chromatography andem mass spectrometry (LCMS/MS) has been the technique of choice throughout the main part of this work, due to the high sensitivity and selectivity requirements.

Generación automática de VHDL mediante Simulink HDL coder y aplicación al procesado de señales

Gordon E. Moore, co-fundador de Intel, predijo en una publicación del año 1965 que aproximadamente cada dos años se duplicaría el número de transistores presentes en un circuito integrado, debido a las cada vez mejores tecnologías presentes en el proceso de elaboración. A esta ley se la conoce como Ley de Moore y su cumplimiento se ha podido constatar hasta hoy en día. Gracias a ello, con el paso del tiempo cada vez se presentan en el mercado circuitos integrados más potentes, con mayores prestaciones para realizar tareas cada vez más complejas. Un tipo de circuitos integrados que han podido evolucionar de forma importante por dicho motivo, son los dispositivos de lógica programable, circuitos integrados que permiten implementar sobre ellos las funciones lógicas que desee implementar el usuario. Hasta hace no muchos años, dichos dispositivos eran capaces de implementar circuitos compuestos por unas pocas funciones lógicas, pero gracias al proceso de miniaturización predicho por la Ley de Moore, hoy en día son capaces de implementar circuitos tan complejos como puede ser un microprocesador; dichos dispositivos reciben el nombre de FPGA, siglas de Field Programmable Gate Array. Debido a la mayor capacidad y por lo tanto a diseños más complejos implementados sobre las FPGA, en los últimos años han aparecido herramientas cuyo objetivo es hacer más fácil el proceso de ingeniería dentro de un desarrollo en este tipo de dispositivos, como es la herramienta HDL Coder de la compañía MathWorks, creadores también Matlab y Simulink, unas potentes herramientas usadas ampliamente en diferentes ramas de la ingeniería. El presente proyecto tiene como objetivo evaluar el uso de dicha herramienta para el procesado digital de señales, usando para ello una FPGA Cyclone II de la casa Altera. Para ello, se empezará analizando la herramienta escogida comparándola con herramientas de la misma índole, para a continuación seleccionar una aplicación de procesado digital de señal a implementar. Tras diseñar e implementar la aplicación escogida, se deberá simular en PC para finalmente integrarla en la placa de evaluación seleccionada y comprobar su correcto funcionamiento. Tras analizar los resultados de la aplicación de implementada, concretamente un analizador de la frecuencia fundamental de una señal de audio, se ha comprobado que la herramienta HDL Coder, es adecuada para este tipo de desarrollos, facilitando enormemente los procesos tanto de implementación como de validación gracias al mayor nivel de abstracción que aporta.

LC-MS based methods for phytosterols determination in oenological matrices. Targeted metabolomic study of Rioja grape varieties

221 p.+ anexos

Human LDL Structural Diversity Studied by IR Spectroscopy

Lipoproteins are responsible for cholesterol traffic in humans. Low density lipoprotein (LDL) delivers cholesterol from liver to peripheral tissues. A misleading delivery can lead to the formation of atherosclerotic plaques. LDL has a single protein, apoB-100, that binds to a specific receptor. It is known that the failure associated with a deficient protein-receptor binding leads to plaque formation. ApoB-100 is a large single lipid-associated polypeptide difficulting the study of its structure. IR spectroscopy is a technique suitable to follow the different conformational changes produced in apoB-100 because it is not affected by the size of the protein or the turbidity of the sample. We have analyzed LDL spectra of different individuals and shown that, even if there are not big structural changes, a different pattern in the intensity of the band located around 1617 cm 21 related with strands embedded in the lipid monolayer, can be associated with a different conformational rearrangement that could affect to a protein interacting region with the receptor.

Adenylate Cyclase Toxin promotes bacterial internalisation into non phagocytic cells

Bordetella pertussis causes whooping cough, a respiratory infectious disease that is the fifth largest cause of vaccine-preventable death in infants. Though historically considered an extracellular pathogen, this bacterium has been detected both in vitro and in vivo inside phagocytic and non-phagocytic cells. However the precise mechanism used by B. pertussis for cell entry, or the putative bacterial factors involved, are not fully elucidated. Here we find that adenylate cyclase toxin (ACT), one of the important toxins of B. pertussis, is sufficient to promote bacterial internalisation into non-phagocytic cells. After characterization of the entry route we show that uptake of "toxin-coated bacteria" proceeds via a clathrin-independent, caveolae-dependent entry pathway, allowing the internalised bacteria to survive within the cells. Intracellular bacteria were found inside non-acidic endosomes with high sphingomyelin and cholesterol content, or "free" in the cytosol of the invaded cells, suggesting that the ACT-induced bacterial uptake may not proceed through formation of late endolysosomes. Activation of Tyr kinases and toxin-induced Ca2+-influx are essential for the entry process. We hypothesize that B. pertussis might use ACT to activate the endocytic machinery of non-phagocytic cells and gain entry into these cells, in this way evading the host immune system.