Topographical and mechanical characterization of living eukaryotic cells on opaque substrates: development of a general procedure and its application to the study of non-adherent lymphocytes

The mechanical behavior of living murine T-lymphocytes was assessed by atomic force microscopy (AFM). A robust experimental procedure was developed to overcome some features of lymphocytes, in particular their spherical shape and non-adherent character. The procedure included the immobilization of the lymphocytes on amine-functionalized substrates, the use of hydrodynamic effects on the deflection of the AFM cantilever to monitor the approaching, and the use of the jumping mode for obtaining the images. Indentation curves were analyzed according to Hertz's model for contact mechanics. The calculated values of the elastic modulus are consistent both when considering the results obtained from a single lymphocyte and when comparing the curves recorded from cells of different specimens

Medidas de propiedades dieléctricas de materiales de construcción mediante elipsometría

A set of measurements of electromagnetic properties of building materials is presented in this work. The method is based on the measurement of the polarization state of the reflected signal from the material under study at a fixed angle of incidence. From the measured data, by using the Fresnel equations, it has been obtained the dielectric constant. Measurements were done by using two horn antennas at the frequency of 9 GHz. The obtained results are compared with the free space reflexion and transmission Fresnel method and other reflection methods based on a conductor waveguide. The method explained in this work can be used for other type of materials and its main advantage is the non-destructive character and the ease implementation.

Desarrollo de un procedimiento para la observación y caracterización mecánica de células sobre sustratos opacos mediante microscopía de fuerzas atómicas : aplicación al estudio de linfocitos

Las células en los tejidos biológicos están continuamente sometidas a estímulos físicos tales como la presión hidrostática y esfuerzos de tracción, compresión o cortante, entre otros. La importancia de los estímulos mecánicos en el comportamiento de las células se ha reconocido recientemente al comprobarse cómo la naturaleza de estas fuerzas puede cambiar en patologías tales como las enfermedades vasculares o el cáncer. En respuesta a estos cambios, las células reaccionan modificando desde su forma o aspecto hasta su ciclo celular. Consecuentemente, el interés por el comportamiento mecánico de las células ha experimentado un auge creciente que ha requerido el desarrollo de varias técnicas de caracterización. En este contexto, se puede afirmar que una de las técnicas que ha irrumpido con más fuerza en esta nueva área, situada entre el mundo biológico y el físico, es la microscopía de fuerza atómica. En esta Tesis se ha abordado el estudio mediante microscopía de fuerza atómica de linfocitos de ratón que constituyen un linaje celular especialmente difícil de caracterizar mediante esta técnica por su tamaño y naturaleza no adherente. Los linfocitos, como actores fundamentales del sistema inmune, tienen gran importancia en la determinación de la respuesta que un organismo desencadena ante la presencia de un biomaterial. Bajo esta premisa, y como condición previa a la caracterización de los linfocitos, ha sido necesario el desarrollo de una metodología robusta y de amplia aplicabilidad que permita el estudio de células sobre biomateriales. Finalmente y con el objetivo de correlacionar el comportamiento mecánico de los linfocitos con alguna característica fisiológica relevante, se ha analizado la hipótesis de que el comportamiento mecánico pueda ser utilizado como marcador de la edad biológica. Consecuentemente se ha abordado el estudio del comportamiento mecánico de los linfocitos clasificados por grupos de edad, de manera que se han obtenido los primeros resultados que indican cómo puede manifestarse el proceso de inmunosenescencia -depresión del sistema inmune relacionada con el envejecimiento- en el comportamiento mecánico de las células del sistema inmune. Cells within tissues are continuously exposed to physical forces including hydrostatic pressure, shear stress, and compression and tension forces. The relevance of these mechanical stimuli has recently been recognised by different works in which significant changes were observed in these forces when they were measued in individuals affected by cardiovasvular diseases or cancer. Cells may alter their orientation, shape, internal constitution, contract, migrate, adhere, modify the synthesis and degradation of extracellular constituents, or even their life cycle in response to perturbations in their mechanical environment. As a consequence of this, the attention in cell mechanical behavior has undergone a significant thrust and novel techniques have been developed. In this context, atomic force microscopy has become a basic tool for the progress of this field. In this Thesis, the mechanical behavior of living murine T-lymphocytes was assessed by atomic force microscopy. Lymphocytes play a main role in the immune system of the individual and, consequently, in the immune response triggered by the presence of a biomaterial. The observation and characterization of the lymphocytes required the development of a robust experimental procedure that allowed overcoming the difficulties related to the analysis of this cell lineage, in particular their relatively large size and non-adherent character. These procedures could be easily transferred to other non-adherent cell lineages. Finally, to check the viability of developed method, we study the lymphocyte mechanical behavior as a function of the murine ageing. The obtained data represent a first step in the knowledge about how mechanical stimuli can affect the age-dependent decrease in immunological competence, i.e., the immunosenescence.

Especificación del comportamiento de un avatar en un mundo virtual mediante una ontología

El presente trabajo describe la construcción de una aplicación que controla a un Non Player Character (NPC), en un mundo virtual. La aplicación desarrollada, que tiene como nombre BotManager, realiza dos tareas fundamentales: 1) conectarse al repositorio de conocimiento, que en esta implementación es una ontología expresada en OWL, para obtener las acciones que debe realizar el NPC dentro del mundo virtual; y 2) ordenar al NPC que realice estas acciones en un mundo virtual creado con la plataforma OpenSimulator. BotManager puede tener variadas aplicaciones, por lo tanto puede ser usada como complemento en mundos virtuales aplicados a la educación, simulación, ocio, etc. Ahora bien, la principal razón que motivó el desarrollo del BotManager fue la de crear un sistema de demostración automática de tareas en un mundo virtual destinado a la educación/ entrenamiento. De esta forma, un Sistema Inteligente de Tutoría integrado con un mundo virtual podría demostrar paso a paso a un estudiante cómo realizar una tarea en el mundo virtual. La ontología que lee el BotManager extiende la ontología propuesta en la tesis “Una propuesta de modelado del estudiante basada en ontologías y diagnóstico pedagógico-cognitivo no monótono” de Julia Parraga en el 2011 (Ontología de Julia). La construcción y las pruebas del BotManager se llevaron a cabo en tres etapas: 1) creación de la Ontología de Acciones del NPC que extiende la Ontología de Julia; 2) diseño e implementación de la aplicación en C# que lee la ontología que contiene el plan de acción del NPC, y ordena al NPC realizar las acciones en el mundo virtual; y 3) pruebas de la aplicación con la práctica “preparación de una taza de cafe”, que es parte de un Laboratorio Virtual de Biotecnología. El BotManager se ha diseñado como una aplicación cliente que se conecta a un servidor de Open- Simulator. Por lo tanto, puede ejecutarse en una máquina distinta a la del servidor. Asimismo, en la implementación del BotManager se ha utilizado una librería gratuita denominada LibOpenMetaverse que permite controlar un NPC de forma remota.---ABSTRACT---This paper describes the construction of an application that controls a Non Player Character (NPC), in a virtual world. The application developed, called BotManager, performs two main tasks: 1) the connection to the repository of knowledge, which in this implementation is an ontology expressed in OWL, and retrieving the actions to be performed by the NPC within the virtual world; and 2) commanding the NPC to perform these actions in a virtual world created with the OpenSimulator platform. BotManager can have diverse applications, therefore it can be used as a complement in virtual worlds applied to education, simulation, entertainment, etc. However, the main reason behind the development of BotManager was to create an automatic demonstration of tasks in a virtual world for education / training. Thus, a virtual world integrated with an Intelligent Tutoring Systems could demonstrate step by step to a student how to perform a task in the virtual world. The ontology used by the BotManager extends ontology proposed in the thesis “A proposal for modeling ontologies based student and not monotonous teaching-cognitive diagnosis” by Julia Parraga in 2011 (Julia’s Ontology). Construction and testing of BotManager were conducted in three stages: 1) creation of the NPC Actions Ontology by extending the Julia’s Ontology; 2) design and implementation of the application in C# that reads the ontology containing the plan of action of the NPC, and commands the NPC to perform the read plan in the virtual world; and 3) testing of the application with the practice “preparing a cup of coffee”, which is part of a Virtual Laboratory of Biotechnology. The BotManager has been designed as a client application that connects to an OpenSimulator server. Therefore, it can run on a different machine to the server. To implement the BotManager we have used a free library called libopenmetaverse that allows us to control a NPC remotely.

Non-uniform target illumination in deflagration regime : thermal smoothing

Thermal smoothing in the plasma ablated from a laser target under weakly nonuniform irradiation is analyzed, assuming absorption at nc and a deflagration regime (conduction restricted to a thin quasisteady layer next to the target). Magnetic generation effects are included and found to be weak. Differences from results available in the literature are explained; the importance of the character of the underdense flow at uniform irradiation is emphasized.

Hexagons and spiral defect chaos in non-Boussinesq convection at low Prandtl numbers

We study the stability and dynamics of non-Boussinesq convection in pure gases ?CO2 and SF6? with Prandtl numbers near Pr? 1 and in a H2-Xe mixture with Pr= 0.17. Focusing on the strongly nonlinear regime we employ Galerkin stability analyses and direct numerical simulations of the Navier-Stokes equations. For Pr ? 1 and intermediate non-Boussinesq effects we find reentrance of stable hexagons as the Rayleigh number is increased. For stronger non-Boussinesq effects the usual, transverse side-band instability is superseded by a longitudinal side-band instability. Moreover, the hexagons do not exhibit any amplitude instability to rolls. Seemingly, this result contradicts the experimentally observed transition from hexagons to rolls. We resolve this discrepancy by including the effect of the lateral walls. Non-Boussinesq effects modify the spiral defect chaos observed for larger Rayleigh numbers. For convection in SF6 we find that non-Boussinesq effects strongly increase the number of small, compact convection cells and with it enhance the cellular character of the patterns. In H2-Xe, closer to threshold, we find instead an enhanced tendency toward roll-like structures. In both cases the number of spirals and of targetlike components is reduced. We quantify these effects using recently developed diagnostics of the geometric properties of the patterns.