Different endocytotic uptake mechanisms for nanoparticles in epithelial cells and macrophages.

Precise knowledge regarding cellular uptake of nanoparticles is of great importance for future biomedical applications. Four different endocytotic uptake mechanisms, that is, phagocytosis, macropinocytosis, clathrin- and caveolin-mediated endocytosis, were investigated using a mouse macrophage (J774A.1) and a human alveolar epithelial type II cell line (A549). In order to deduce the involved pathway in nanoparticle uptake, selected inhibitors specific for one of the endocytotic pathways were optimized regarding concentration and incubation time in combination with fluorescently tagged marker proteins. Qualitative immunolocalization showed that J774A.1 cells highly expressed the lipid raft-related protein flotillin-1 and clathrin heavy chain, however, no caveolin-1. A549 cells expressed clathrin heavy chain and caveolin-1, but no flotillin-1 uptake-related proteins. Our data revealed an impeded uptake of 40 nm polystyrene nanoparticles by J774A.1 macrophages when actin polymerization and clathrin-coated pit formation was blocked. From this result, it is suggested that macropinocytosis and phagocytosis, as well as clathrin-mediated endocytosis, play a crucial role. The uptake of 40 nm nanoparticles in alveolar epithelial A549 cells was inhibited after depletion of cholesterol in the plasma membrane (preventing caveolin-mediated endocytosis) and inhibition of clathrin-coated vesicles (preventing clathrin-mediated endocytosis). Our data showed that a combination of several distinguishable endocytotic uptake mechanisms are involved in the uptake of 40 nm polystyrene nanoparticles in both the macrophage and epithelial cell line.

Dielectrophoresis-based analyte separation and analysis

Dielectrophoresis—the tendency of a material of high dielectric permittivity to migrate in an electrical field gradient to a region of maximum field strength—provides an ideal motive force for manipulating small volumes of biological analytes in microfluidic microsystems. The work described in this thesis was based on the hypothesis that dielectrophoresis could be exploited to provide high-resolution cell separations in microsystems as well as a means for the electrically-controllable manipulation of solid supports for molecular analysis. To this end, a dielectrophoretic/gravitational field-flow-fractionation (DEP/G-FFF) system was developed and the separation performance evaluated using various types and sizes of polystyrene microspheres as model particles. It was shown that separation of the polystyrene beads was based on the differences in their effective dielectrophoretic properties. The ability of an improved DEP/G-FFF system to separate genetically identical, but phenotypically dissimilar cell types was demonstrated using mixtures of 6m2 mutant rat kidney cells grown under transforming and non-transforming culture conditions. Additionally, a panel of engineered dielectric microspheres was designed with specific, predetermined dielectrophoretic properties such that their dielectrophoretic behaviors would be controllable and predictable. The fabrication method involved the use of gold-coated polystyrene microsphere cores coated with a self-assembled monolayer of alkanethiol and, optionally, a self-assembled monolayer of phospholipid to form a thin-insulating-shell-over-conductive-interior structure. The successful development of the DEP/G-FFF separation system and the dielectrically engineered microspheres provides proof-of-principle demonstrations of enabling dielectrophoresis-based microsystem technology that should provide powerful new methods for the manipulation, separation and identification of analytes in many diverse fields. ^

Biological Signatures of Vaccine Responses

The set of host- and pathogen-specific molecular features of a disease comprise its “signature”. We hypothesize that biological signatures enables distinctions between vaccinated vs. infected individuals. In our research, using porcine samples, protocols were developed that could also be used to identify biological signatures of human disease. Different classes of molecular features will be tested during this project, including indicators of basic immune capacity, which are being studied at this instance. These indicators of basic immune response such as porcine cytokines and antibodies were validated using Enzyme-linked immunosorbent assay (ELISA). This is an established method that detects antigens by their interaction with a specific antibody coupled to a polystyrene substrate. Serum from naïve and vaccinated pigs was tested for the presence of cytokines. We were able to differentiate the presence of porcine IL-6 in normal porcine serum with or without added porcine IL-6 by ELISA. In addition, four different cytokines were spotted on a grating-coupled surface plasmon resonance imaging system (GCSPRI) chip and antibody specific for IL-8 was run over the chip. Only the presence of IL-8 was detected; therefore, there was no cross-reactivity in this combination of antigens and antibodies. This system uses a multiplexed sensor chip to identify components of a sample run over it. The detection is accomplished by the change in refractive index caused by the interaction between the antibody spotted on the sensor chip and the antigen present in the sample. As the multiplexed GCSPRI is developed, we will need to optimize both sensitivity and specificity, minimizing the potential for cross-reactivity between individual analytes. The next step in this project is to increase the sensitivity of detection of the analytes. Currently, we are using two different antibodies (that recognize a different part of the antigen) to amplify the signal emitted by the interaction of antibody with its cognate antigen. The development of this sensor chip would not only allow to detect FMD virus, but also to differentiate between infected and vaccinated individuals, on location. Furthermore, the diagnosis of other diseases could be done with increased accuracy, and in less time due to the microarray approach.

Development and Implementation of the Use of Optically Stimulated Luminescent Detectors in the Radiological Physics Center Anthropomorphic Quality Assurance Phantoms

The Radiological Physics Center (RPC) uses both on-site and remote reviews to credential institutions for participation in clinical trials. Anthropomorphic quality assurance (QA) phantoms are one tool the RPC uses to remotely audit institutions, which include thermoluminescent dosimeters (TLDs) and radiochromic film. The RPC desires to switch from TLD as the absolute dosimeter in the phantoms, to optically stimulated luminescent dosimeters (OSLDs), but a problem lies in the angular dependence exhibited by the OSLD. The purpose of this study was to characterize the angular dependence of OSLD and establish a correction factor if necessary, to provide accurate dosimetric measurements as a replacement for TLD in the QA phantoms. A 10 cm diameter high-impact polystyrene spherical phantom was designed and constructed to hold an OSLD to study the angular response of the dosimeter under the simplest of circumstances for both coplanar and non-coplanar treatment deliveries. OSLD were irradiated in the spherical phantom, and the responses of the dosimeter from edge-on angles were normalized to the response when irradiated with the beam incident normally on the surface of the dosimeter. The average normalized response was used to establish an angular correction factor for 6 MV and 18 coplanar treatments, and for 6 MV non-coplanar treatments specific to CyberKnife. The RPC pelvic phantom dosimetry insert was modified to hold OSLD, in addition to the TLD, adjacent to the planes of film. Treatment plans of increasing angular beam delivery were developed, three in Pinnacle v9.0 (4-field box, IMRT, and VMAT) and one in Accuray’s MultiPlan v3.5.3 (CyberKnife). The plans were delivered to the pelvic phantom containing both TLD and OSLD in the target volume. The pelvic phantom was also sent to two institutions to be irradiated as trials, one delivering IMRT, and the other a CyberKnife treatment. For the IMRT deliveries and the two institution trials, the phantom also included film in the sagittal and coronal planes. The doses measured from the TLD and OSLD were calculated for each irradiation, and the angular correction factors established from the spherical phantom irradiations were applied to the OSLD dose. The ratio of the TLD dose to the angular corrected OSLD dose was calculated for each irradiation. The corrected OSLD dose was found to be within 1% of the TLD measured dose for all irradiations, with the exception of the in-house CyberKnife deliveries. The films were normalized to both TLD measured dose and the corrected OSLD dose. Dose profiles were obtained and gamma analysis was performed using a 7%/4 mm criteria, to compare the ability of the OSLD, when corrected for the angular dependence, to provide equivalent results to TLD. The results of this study indicate that the OSLD can effectively be used as a replacement for TLD in the RPC’s anthropomorphic QA phantoms for coplanar treatment deliveries when a correction is applied for the dosimeter’s angular dependence.

An approach for particle sinking velocity measurements in the 3-400 µm size range and considerations on the effect of temperature on sinking rates

The flux of organic particles below the mixed layer is one major pathway of carbon from the surface into the deep ocean. The magnitude of this export flux depends on two major processes-remineralization rates and sinking velocities. Here, we present an efficient method to measure sinking velocities of particles in the size range from approximately 3-400 µm by means of video microscopy (FlowCAM®). The method allows rapid measurement and automated analysis of mixed samples and was tested with polystyrene beads, different phytoplankton species, and sediment trap material. Sinking velocities of polystyrene beads were close to theoretical values calculated from Stokes' Law. Sinking velocities of the investigated phytoplankton species were in reasonable agreement with published literature values and sinking velocities of material collected in sediment trap increased with particle size. Temperature had a strong effect on sinking velocities due to its influence on seawater viscosity and density. An increase in 9 °C led to a measured increase in sinking velocities of 40 %. According to this temperature effect, an average temperature increase in 2 °C as projected for the sea surface by the end of this century could increase sinking velocities by about 6 % which might have feedbacks on carbon export into the deep ocean.

New applications of cork in building

Several types of prefabricated elements for partitions and extrados, which improve the similar systems presently in the market, are presented in this research. These elements, manufactured with cork and lightened plaster, can be classified in two well defined groups. The first group is formed by lightened plaster panels for partitions of dimensions 60 x 265 cm (width, height) and of 7cm and 10 cm thickness. In the second group, panels for extrados with the same dimensions are included but with a thickness of 9.5 and 5 cm, including the 3 cm thickness of the incorporated high density expanded polystyrene sheet.

Inmovilización de TiO2 sobre polímeros transparentes en el UV-A para la eliminación fotocatalítica de tricloroetileno en aire

El gran desarrollo industrial y demográfico de las últimas décadas ha dado lugar a un consumo crecientemente insostenible de energía y materias primas, que influye negativamente en el ambiente por la gran cantidad de contaminantes generados. Entre las emisiones tienen gran importancia los compuestos orgánicos volátiles (COV), y entre ellos los compuestos halogenados como el tricloroetileno, debido a su elevada toxicidad y resistencia a la degradación. Las tecnologías generalmente empleadas para la degradación de estos compuestos presentan inconvenientes derivados de la generación de productos tóxicos intermedios o su elevado coste. Dentro de los procesos avanzados de oxidación (Advanced Oxidation Processes AOP), la fotocatálisis resulta una técnica atractiva e innovadora de interés creciente en su aplicación para la eliminación de multitud de compuestos orgánicos e inorgánicos, y se ha revelado como una tecnología efectiva en la eliminación de compuestos orgánicos volátiles clorados como el tricloroetileno. Además, al poder aprovechar la luz solar como fuente de radiación UV permite una reducción significativa de costes energéticos y de operación. Los semiconductores más adecuados para su empleo como fotocatalizadores con aprovechamiento de la luz solar son aquellos que tienen una banda de energía comparable a la de los fotones de luz visible o, en su defecto, de luz ultravioleta A (Eg < 3,5 eV), siendo el más empleado el dióxido de titanio (TiO2). El objetivo principal de este trabajo es el estudio de polímeros orgánicos comerciales como soporte para el TiO2 en fotocatálisis heterogénea y su ensayo para la eliminación de tricloroetileno en aire. Para ello, se han evaluado sus propiedades ópticas y su resistencia a la fotodegradación, y se ha optimizado la fijación del fotocatalizador para conseguir un recubrimiento homogéneo, duradero y con elevada actividad fotocatalítica en diversas condiciones de operación. Los materiales plásticos ensayados fueron el polietileno (PE), copolímero de etil vinil acetato con distintos aditivos (EVA, EVA-H y EVA-SH), polipropileno (PP), polimetil (metacrilato) fabricado en colada y extrusión (PMMA-C y PMMA-E), policarbonato compacto y celular (PC-C y PC-Ce), polivinilo rígido y flexible (PVC-R y PVC-F), poliestireno (PS) y poliésteres (PET y PETG). En base a sus propiedades ópticas se seleccionaron el PP, PS, PMMA-C, EVA-SH y PVC-R, los cuales mostraron un valor de transmitancia superior al 80% en el entorno de la región estudiada (λ=365nm). Para la síntesis del fotocatalizador se empleó la tecnología sol-gel y la impregnación multicapa de los polímeros seleccionados por el método de dip-coating con secado intermedio a temperaturas moderadas. Con el fin de evaluar el envejecimiento de los polímeros bajo la radiación UV, y el efecto sobre éste del recubrimiento fotoactivo, se realizó un estudio en una cámara de exposición a la luz solar durante 150 días, evaluándose la resistencia química y la resistencia mecánica. Los resultados de espectroscopía infrarroja y del test de tracción tras el envejecimiento revelaron una mayor resistencia del PMMA y una degradación mayor en el PS, PVC-R y EVA SH, con una apreciable pérdida del recubrimiento en todos los polímeros. Los fotocatalizadores preparados sobre soportes sin tratamiento y con tres capas de óxido de titanio mostraron mejores resultados de actividad con PMMA-C, PET y PS, con buenos resultados de mineralización. Para conseguir una mayor y mejor fijación de la película al soporte se realizaron tratamientos químicos abrasivos con H2SO4 y NaOH y tratamientos de funcionalización superficial por tecnología de plasma a presión atmosférica (APP) y a baja presión (LPP). Con los tratamientos de plasma se consiguió una excelente mojabilidad de los soportes, que dio lugar a una distribución uniforme y más abundante del fotocatalizador, mientras que con los tratamientos químicos no se obtuvo una mejora significativa. Asimismo, se prepararon fotocatalizadores con una capa previa de dióxido de silicio con la intervención de surfactantes (PDDA-SiO2-3TiO2 y SiO2FC-3TiO2), consiguiéndose buenas propiedades de la película en todos los casos. Los mejores resultados de actividad con tratamiento LPP y tres capas de TiO2 se lograron con PMMA-C (91% de conversión a 30 ppm de TCE y caudal 200 ml·min-1) mejorando significativamente también la actividad fotocatalítica en PVC-R y PS. Sin embargo, el material más activo de todos los ensayados fue el PMMA-C con el recubrimiento SiO2FC-3TiO2, logrando el mejor grado de mineralización, del 45%, y una velocidad de 1,89 x 10-6 mol· m-2 · s-1, que dio lugar a la eliminación del 100 % del tricloroetileno en las condiciones anteriormente descritas. A modo comparativo se realizaron ensayos de actividad con otro contaminante orgánico tipo, el formaldehído, cuya degradación fotocatalítica fue también excelente (100% de conversión y 80% de mineralización con 24 ppm de HCHO en un caudal de aire seco de 200 ml·min-1). Los buenos resultados de actividad obtenidos confirman las enormes posibilidades que ofrecen los polímeros transparentes en el UV-A como soportes del dióxido de titanio para la eliminación fotocatalítica de contaminantes en aire. ABSTRACT The great industrial and demographic development of recent decades has led to an unsustainable increase of energy and raw materials consumption that negatively affects the environment due to the large amount of waste and pollutants generated. Between emissions generated organic compounds (VOCs), specially the halogenated ones such as trichloroethylene, are particularly important due to its high toxicity and resistance to degradation. The technologies generally used for the degradation of these compounds have serious inconveniences due to the generation of toxic intermediates turn creating the problem of disposal besides the high cost. Among the advanced oxidation processes (AOP), photocatalysis is an attractive and innovative technique with growing interest in its application for the removal of many organic and inorganic compounds, and has emerged as an effective technology in eliminating chlorinated organic compounds such as trichloroethylene. In addition, as it allows the use of sunlight as a source of UV radiation there is a significant reduction of energy costs and operation. Semiconductors suitable to be used as photocatalyst activated by sunlight are those having an energy band comparable to that of the visible or UV-A light (Eg <3,5 eV), being titanium dioxide (TiO2), the most widely used. The main objective of this study is the test of commercial organic polymers as supports for TiO2 to be applied in heterogeneous photocatalysis and its assay for removing trichloroethylene in air. To accomplish that, its optical properties and resistance to photooxidation have been evaluated, and different operating conditions have been tested in order to optimize the fixation of the photocatalyst to obtain a homogeneous coating, with durable and high photocatalytic activity. The plastic materials tested were: polyethylene (PE), ethyl vinyl acetace copolymers with different additives (EVA, EVA-H and EVA -SH), polypropylene (PP), poly methyl (methacrylate) manufactured by sheet moulding and extrusion (PMMA-C and PMMA-E), compact and cellular polycarbonates (PC-C PC-Ce), rigid and flexible polyvinyl chloride (PVC-R and PVC-F), polystyrene (PS) and polyesters (PET and PETG). On the basis of their optical properties PP, PS, PMMA-C, EVA-SH and PVC-R were selected, as they showed a transmittance value greater than 80% in the range of the studied region (λ = 365nm). For the synthesis of the photocatalyst sol-gel technology was employed with multilayers impregnation of the polymers selected by dip-coating, with intermediate TiO2 drying at moderate temperatures. To evaluate the polymers aging due to UV radiation, and the effect of photoactive coating thereon, a study in an sunlight exposure chamber for 150 days was performed, evaluating the chemical resistance and the mechanical strength. The results of infrared spectroscopy and tensile stress test after aging showed the PMMA is the most resistant sample, but a greater degradation in PS, PVC-R and EVA SH, with a visible loss of the coating in all the polymers tested. The photocatalysts prepared on the untreated substrates with three layers of TiO2 showed better activity results when PMMA-C, PET and PS where used. To achieve greater and better fixation of the film to the support, chemical abrasive treatments, with H2SO4 and NaOH, as well as surface functionalization treatments with atmospheric pressure plasma (APP) and low pressure plasma (LPP) technologies were performed. The plasma treatment showed the best results, with an excellent wettability of the substrates that lead to a better and uniform distribution of the photocatalyst compared to the chemical treatments tested, in which no significant improvement was obtained. Also photocatalysts were prepared with the a silicon dioxide previous layer with the help of surfactants (SiO2- 3TiO2 PDDA-and-3TiO2 SiO2FC), obtaining good properties of the film in all cases. The best activity results for LPP-treated samples with three layers of TiO2 were achieved with PMMA-C (91% conversion, in conditions of 30 ppm of TCE and 200 ml·min-1 air flow rate), with a significant improvement of the photocatalytic activity in PVC-R and PS samples too. However, among all the materials assayed, PMMA-C with SiO2FC-3TiO2 coating was the most active one, achieving the highest mineralization grade (45%) and a reaction rate of 1,89 x 10-6 mol· m-2 · s-1, with total trichloroethylene elimination in the same conditions. As a comparative assay, an activity test was also performed with another typical organic contaminant, formaldehyde, also with good results (100% conversion with 24 ppm of HCHO and 200 ml·min-1 gas flow rate). The good activity results obtained in this study confirm the great potential of organic polymers which are transparent in the UV-A as supports for titanium dioxide for photocatalytic removal of air organic pollutants.

Polypropylene/glass fiber hierarchical composites incorporating inorganic fullerene-like nanoparticles for advanced technological applications

Novel isotactic polypropylene (iPP)/glass fiber (GF) laminates reinforced with inorganic fullerene-like tungsten disulfide (IF-WS2) nanoparticles as environmentally friendly fillers have been successfully fabricated by simple melt-blending and fiber impregnation in a hot-press without the addition of any compatibilizer. The influence of IF-WS2 concentration on the morphology, viscosity. and thermal and mechanical behavior of the hierarchical composites has been investigated. Results revealed an unprecedented 62 °C increase in the degradation temperature of iPP/GF upon addition of only 4.0 wt % IF-WS2. The coexistence of both micro- and nanoscale fillers resulted in synergistic effects on enhancing the stiffness, strength, crystallinity, thermal stability, glass transition (Tg) and heat distortion temperature (HDT) of the matrix. The approach used in this work is an efficient, versatile, scalable and economic strategy to improve the mechanical and thermal behavior of GF-reinforced thermoplastics with a view to extend their use in advanced technological applications. This new type of composite materials shows great potential to improve the efficiency and sustainability of many forms of transport.

Evaluating the reinforcement of inorganic fullerene-like nanoparticles in thermoplastic matrices by depth-sensing indentation

The reinforcing effect of inorganic fullerene-like tungsten disulfide (IF-WS2) nanoparticles in two different polymer matrices, isotactic polypropylene (iPP) and polyphenylene sulfide (PPS), has been investigated by means of dynamic depth-sensing indentation. The hardness and elastic modulus enhancement upon filler addition is analyzed in terms of two main contributions: changes in the polymer matrix nanostructure and intrinsic properties of the filler including matrix-particle load transfer. It is found that the latter mainly determines the overall mechanical improvement, whereas the nanostructural changes induced in the polymer matrix only contribute to a minor extent. Important differences are suggested between the mechanisms of deformation in the two nanocomposites, resulting in a moderate mechanical enhancement in case of iPP (20% for a filler loading of 1%), and a remarkable hardness increase in case of PPS (60% for the same filler content). The nature of the polymer amorphous phase, whether in the glassy or rubbery state, seems to play here an important role. Finally, nanoindentation and dynamic mechanical analysis measurements are compared and discussed in terms of the different directionality of the stresses applied.

Influencia de las condiciones de ejecución en la resistencia de anclajes en convencional y autocompactante

A través de los años las estructuras de hormigón armado han ido aumentando su cuota de mercado, sustituyendo a las estructuras de fábrica de piedra o ladrillo y restándole participación a las estructuras metálicas. Uno de los primeros problemas que surgieron al ejecutar las estructuras de hormigón armado, era cómo conectar una fase de una estructura de este tipo a una fase posterior o a una modificación posterior. Hasta los años 80-90 las conexiones de una fase de una estructura de hormigón armado, con otra posterior se hacían dejando en la primera fase placas de acero con garrotas embebidas en el hormigón fresco o barras grifadas recubiertas de poliestireno expandido. Una vez endurecido el hormigón se podían conectar nuevas barras, para la siguiente fase mediante soldadura a la placa de la superficie o enderezando las barras grifadas, para embeberlas en el hormigón fresco de la fase siguiente. Estos sistemas requerían conocer la existencia y alcance de la fase posterior antes de hormigonar la fase previa. Además requerían un replanteo muy exacto y complejo de los elementos de conexión. Otro problema existente en las estructuras de hormigón era la adherencia de un hormigón fresco a un hormigón endurecido previamente, ya que la superficie de contacto de ambos hormigones suponía un punto débil, con una adherencia baja. A partir de los años 80, la industria química de la construcción experimentó un gran avance en el desarrollo de productos capaces de generar una buena adherencia sobre el hormigón endurecido. Este avance tecnológico tenía aplicación tanto en la adherencia del hormigón fresco sobre el hormigón endurecido, como en la adherencia de barras post-instaladas en agujeros de hormigón endurecido. Este sistema se denominó “anclajes adherentes de barras de acero en hormigón endurecido”. La forma genérica de ejecutarlos es hacer una perforación cilíndrica en el soporte de hormigón, con una herramienta especifica como un taladro, limpiar la perforación, llenarla del material adherente y finalmente introducir la barra de acero. Los anclajes adherentes se dividen en anclajes cementosos y anclajes químicos, siendo estos últimos los más habituales, fiables, resistentes y fáciles de ejecutar. El uso del anclaje adherente de barras de acero en hormigón endurecido se ha extendido por todo el espectro productivo, siendo muy habitual tanto en construcción de obras de hormigón armado de obra civil y edificación, como en obras industriales, instalaciones o fijación de elementos. La ejecución de un anclaje de una barra de acero en hormigón endurecido depende de numerosas variables, que en su conjunto, o de forma aislada pueden afectar de forma notable a la resistencia del anclaje. Nos referimos a variables de los anclajes, que a menudo no se consideran tales como la dirección de la perforación, la máquina de perforación y el útil de perforación utilizado, la diferencia de diámetros entre el diámetro del taladro y la barra, el tipo de material de anclaje, la limpieza del taladro, la humedad del soporte, la altura del taladro, etc. La utilización en los últimos años de los hormigones Autocompactables, añade una variable adicional, que hasta ahora apenas ha sido estudiada. En línea con lo apuntado, la presente tesis doctoral tiene como objetivo principal el estudio de las condiciones de ejecución en la resistencia de los anclajes en hormigón convencional y autocompactable. Esta investigación se centra principalmente en la evaluación de la influencia de una serie de variables sobre la resistencia de los anclajes, tanto en hormigón convencional como en un hormigón autocompactable. Para este estudio ha sido necesaria la fabricación de dos soportes de hormigón sobre los cuales desarrollar los ensayos. Uno de los bloques se ha fabricado con hormigón convencional y el otro con hormigón autocompactable. En cada pieza de hormigón se han realizado 174 anclajes con barras de acero, variando los parámetros a estudiar, para obtener resultados de todas las variables consideradas. Los ensayos a realizar en ambos bloques son exactamente iguales, para poder comparar la diferencia entre un anclaje en un soporte de hormigón con vibrado convencional (HVC) y un hormigón autocompactante (HAC). De cada tipo de ensayo deseado se harán dos repeticiones en la misma pieza. El ensayo de arrancamiento de las barras se realizara con un gato hidráulico hueco, con un sistema de instrumentación de lectura y registro de datos en tiempo real. El análisis de los resultados, realizado con una potente herramienta estadística, ha permitido determinar y evaluar numéricamente la influencia de los variables consideradas en la resistencia de los anclajes realizados. Así mismo ha permitido diferenciar los resultados obtenidos en los hormigones convencionales y autocompactantes, tanto desde el punto de vista de la resistencia mecánica, como de las deformaciones sufridas en el arrancamiento. Se define la resistencia mecánica de un anclaje, como la fuerza desarrollada en la dirección de la barra, para hacer su arrancamiento del soporte. De la misma forma se considera desplazamiento, a la separación entre un punto fijo de la barra y otro del soporte, en la dirección de la barra. Dichos puntos se determinan cuando se ha terminado el anclaje, en la intersección de la superficie plana del soporte, con la barra. Las conclusiones obtenidas han permitido establecer qué variables afectan a la ejecución de los anclajes y en qué cuantía lo hacen, así como determinar la diferencia entre los anclajes en hormigón vibrado convencional y hormigón autocompactante, con resultados muy interesantes, que permiten valorar la influencia de dichas variables. Dentro de las conclusiones podemos destacar tres grupos, que denominaremos como de alta influencia, baja influencia y sin influencia. En todos los casos hay que hacer el estudio en términos de carga y de desplazamiento. Podemos considerar como de alta influencia, en términos de carga las variables de máquina de perforación y el material de anclaje. En términos de desplazamiento podemos considerar de alta influencia además de la máquina de perforación y el material de anclaje, el diámetro del taladro, así como la limpieza y humedad del soporte. Podemos considerar de baja influencia, en términos de carga las variables de tipo de hormigón, dirección de perforación, limpieza y humedad del soporte. En términos de desplazamiento podemos considerar de baja influencia el tipo de hormigón y la dirección de perforación. Podemos considerar en el apartado de “sin influencia”, en términos de carga las variables de diámetro de perforación y altura del taladro. En términos de desplazamiento podemos considerar como “sin influencia” la variable de altura del taladro. Podemos afirmar que las diferencias entre los valores de carga aumentan de forma muy importante en términos de desplazamiento. ABSTRACT Over the years the concrete structures have been increasing their market share, replacing the masonry structures of stone or brick and subtracting as well the participation of the metallic structures. One of the first problems encountered in the implementing of the reinforced concrete structures was connecting a phase structure of this type at a later stage or a subsequent amendment. Until the 80s and 90s the connections of one phase of a reinforced concrete structure with a subsequent first phase were done by leaving the steel plates embedded in the fresh concrete using hooks or bent bars coated with expanded polystyrene. Once the concrete had hardened new bars could be connected to the next stage by welding them to the surface plate or by straightening the bent bars to embed them in the fresh concrete of the next phase. These systems required a previous knowledge of the existence and scope of the subsequent phase before concreting the previous one. They also required a very precise and complex rethinking of the connecting elements. Another existing problem in the concrete structures was the adhesion of a fresh concrete to a previously hardened concrete, since the contact surface of both concretes leaded to a weak point with low adherence. Since the 80s, the chemicals construction industry experienced a breakthrough in the development of products that generate a good grip on the concrete. This technological advance had its application both in the grip on one hardened fresh concrete and in the adhesion of bar post-installed in holes of hardened concrete. This system was termed as adherent anchors of steel bars in hardened concrete. The generic way of executing this system is by firstly drilling a cylindrical hole in the concrete support using a specific tool such as a drill. Then, cleaning the bore and filling it with bonding material to lastly, introduce the steel bar. These adherent anchors are divided into cement and chemical anchors, the latter being the most common, reliable, durable and easy to run. The use of adhesive anchor of steel bars in hardened concrete has spread across the production spectrum turning itself into a very common solution in both construction of reinforced concrete civil engineering and construction, and industrial works, installations and fixing elements as well. The execution of an anchor of a steel bar in hardened concrete depends on numerous variables which together or as a single solution may significantly affect the strength of the anchor. We are referring to variables of anchors which are often not considered, such as the diameter difference between the rod and the bore, the drilling system, cleansing of the drill, type of anchor material, the moisture of the substrate, the direction of the drill, the drill’s height, etc. During recent years, the emergence of self-compacting concrete adds an additional variable which has hardly been studied so far. According to mentioned this thesis aims to study the main performance conditions in the resistance of conventional and self-compacting concrete anchors. This research is primarily focused on the evaluation of the influence of several variables on the strength of the anchoring, both in conventional concrete and self-compacting concrete. In order to complete this study it has been required the manufacture of two concrete supports on which to develop the tests. One of the blocks has been manufactured with conventional concrete and the other with self-compacting concrete. A total of 174 steel bar anchors have been made in each one of the concrete pieces varying the studied parameters in order to obtain results for all variables considered. The tests to be performed on both blocks are exactly the same in order to compare the difference between an anchor on a stand with vibrated concrete (HVC) and a self-compacting concrete (SCC). Each type of test required two repetitions in the same piece. The pulling test of the bars was made with a hollow jack and with an instrumentation system for reading and recording data in real time. The use of a powerful statistical tool in the analysis of the results allowed to numerically determine and evaluate the influence of the variables considered in the resistance of the anchors made. It has likewise enabled to differentiate the results obtained in the self-compacting and conventional concretes, from both the outlook of the mechanical strength and the deformations undergone by uprooting. The mechanical strength of an anchor is defined as the strength undergone in a direction of the bar to uproot it from the support. Likewise, the movement is defined as the separation between a fixed point of the bar and a fixed point from the support considering the direction of the bar. These points are only determined once the anchor is finished, with the bar, at the intersection in the flat surface of the support. The conclusions obtained have established which variables affect the execution of the anchors and in what quantity. They have also permitted to determine the difference between the anchors in vibrated concrete and selfcompacting concrete with very interesting results that also allow to assess the influence of these mentioned variables. Three groups are highlighted among the conclusions called high influence, low influence and no influence. In every case is necessary to perform the study in terms of loading and movement. In terms of loading, there are considered as high influence two variables: drilling machinery and anchorage material. In terms of movement, there are considered as high influence the drilling diameter and the cleaning and moisture of the support, besides the drilling machinery and the anchorage material. Variables such as type of concrete, drilling direction and cleaning and moisture of the support are considered of low influence in terms of load. In terms of movement, the type of concrete and the direction of the drilling are considered variables of low influence. Within the no influence section in terms of loading, there are included the diameter of the drilling and the height of the drill. In terms of loading, the height of the drill is considered as a no influence variable. We can affirm that the differences among the loading values increase significantly in terms of movement.

Análisis de la durabilidad de la cubierta plana invertida, a través del estudio de las interacciones e incompatibilidades entre las membranas sintéticas y el poliestireno extrusionado

La cubierta plana invertida se forma, cuando sobre una membrana impermeable se coloca un aislamiento térmico. Estos materiales pueden separarse con capas auxiliares que impiden el contacto directo entre ellos. Muchos de los materiales que forman esta solución constructiva pueden ser polímeros, como lo son algunas membranas impermeables, las capas auxiliares separadoras o el poliestireno extrusionado (XPS). La mayoría de los polímeros son incompatibles entre sí, por lo que en esta tesis se plantea como hipótesis la posibilidad de que se produzcan interacciones e incompatibilidades entre algunos de estos materiales. Por ello se hace una búsqueda bibliográfica y documental de otras investigaciones que pudieran estar relacionadas con el tema, y se estudian y analizan normas y documentación facilitada por fabricantes. Sin embargo, tras consultar toda la bibliografía y documentación que se referencia en esta tesis doctoral, no fue encontrado ningún trabajo de investigación sobre la influencia de la interacción entre los materiales que componen las cubiertas planas invertidas, y de cómo afecta esta a la durabilidad de las mismas. El propósito de esta tesis es el análisis de la durabilidad de la cubierta plana invertida, desde el punto de vista de las interacciones e incompatibilidades que pueden producirse entre los materiales formantes de esta solución constructiva. Además de este objetivo general, se estudian alternativas que puedan prolongar el ciclo de vida de la cubierta plana invertida. Para ello, se desarrolla un plan experimental con el fin de analizar dichas incompatibilidades, y estudiar los factores que las condicionan. Algunos de los resultados obtenidos, muestran que determinadas láminas impermeables utilizadas normalmente para la construcción de cubiertas planas, pueden interactuar con el XPS y sufrir deterioro. El contacto con el mismo, la incorrecta separación, la presión y el calor, son factores determinantes para que se produzca deterioro. Se puede señalar como una de las conclusiones de esta investigación, que la protección térmica que proporciona el XPS a la cubierta plana invertida, puede no ser suficiente (dependiendo del espesor del mismo, y de la ubicación de la cubierta fundamentalmente), para reducir la cantidad de calor que alcanza la línea de contacto entre los materiales, y por tanto paliar las interacciones que se producen. Además, aunque los geotextiles utilizados como capas auxiliares separadoras, en los gramajes por metro cuadrado, recomendados por algunas de las normas reguladoras de este tipo de cubiertas minoran las interacciones, estas siguen produciéndose. ABSTRACT The inverted flat roof is formed from a waterproofing membrane on which the thermal insulation is placed. These materials may be separated with auxiliary layers, which prevent the direct contact between them. Many of the materials forming this constructive solution can be polymers, such as some waterproofing membranes, the auxiliary separating layers, or the extruded polystyrene board (XPS). Most polymers are incompatible, so this thesis hypothesized that interactions and incompatibilities between some of these materials might be possible. Therefore, a literature search, and other documentation that could be related to the topic, are studied and analyzed, as well as, standards and documentation provided by manufacturers. However, after consulting all literature and documents referenced in this dissertation, it was not found any research about the influence of interaction between the materials forming the inverted flat roof, and how this affects to the durability of them. The purpose of this thesis is the analysis of the durability of the inverted flat roof, from the point of view of interactions and incompatibilities may occur between the materials setting up this constructive solution. Along with this general objective, alternatives that can prolong the life cycle of the inverted roof are studied. To get this, an experimental plan is developed, in order to analyze these incompatibilities, and study the factors conditioning them. Some of the results show, that certain normally used waterproofing laminas for building flat roofs, may interact with XPS producing lamina deterioration. Contact, incorrect separation, pressure and heat, are determinant factors for degradation. It can be pointed out, as one of the conclusions of this research, that the thermal protection provided by XPS to the inverted flat roof, cannot be enough (depending on the XPS thickness, and the location of the flat mainly) to reduce the amount of heat that reaches the contact line between materials, and thus to cut down interactions. Furthermore, although geotextiles used as auxiliary separating layers, within the weights per square meter recommended by some of the regulating rules of this type of roofs reduce interactions, these still occur.

Caracterización físico-mecánica del yeso de construcción con adición de residuos de poliestireno extruído y residuos cerámicos

Este trabajo estudia el comportamiento de una matriz de yeso de construcción a la que se le han añadido residuos de construcción y demolición (RCD), residuos de poliestireno extruido (XPS) y residuos cerámicos respectivamente, combinados y en diferentes porcentajes en función del peso del yeso. Los residuos de XPS son producto de una obra en Madrid donde el material fue utilizado como aislamiento térmico y los residuos cerámicos corresponden a trozos de ladrillos toscos encontrados en una obra paralizada en la ciudad de Ávila. Se confeccionaron probetas con porcentajes hasta 3% de XPS y hasta 50% de cerámicos en función del peso del yeso utilizado, como referencia se confeccionaron probetas sin adición de RCD. Fueron ensayadas en laboratorio y se determinaron las características físicas y mecánicas de las mismas. Tras un análisis comparativo se evidencian que la adición de residuos de XPS y residuos cerámicos en conjunto disminuye la densidad seca del material y la absorción de agua por capilaridad, en algunos casos disminuye la conductividad térmica y aumenta la dureza superficial. ABSTRACT: This paper studies the behavior of a building gypsum matrix to which have been added Construction and Demolition waste (CDW), residues of extruded polystyrene (XPS) and ceramic waste respectively, and combined in different percentages depending on the weight of gypsum. XPS waste are the product of a work in Madrid where the material was used as thermal insulation and ceramic waste correspond to pieces of rough bricks found in a paralyzed work in the city of Avila. Specimens were prepared with percentages up to 3% of XPS and up to 50% depending on the weight ceramic gypsum used as reference samples were prepared without addition of CDW. They were tested in laboratory and determined the physical and mechanical characteristics thereof. After a comparative analysis they show that the addition of ceramic waste and waste XPS decreases together dry material density and water absorption by capillary action, in some cases the thermal conductivity decreases and increases surface hardness.

Stimulation of neurite outgrowth using an electrically conducting polymer

Damage to peripheral nerves often cannot be repaired by the juxtaposition of the severed nerve ends. Surgeons have typically used autologous nerve grafts, which have several drawbacks including the need for multiple surgical procedures and loss of function at the donor site. As an alternative, the use of nerve guidance channels to bridge the gap between severed nerve ends is being explored. In this paper, the electrically conductive polymer—oxidized polypyrrole (PP)—has been evaluated for use as a substrate to enhance nerve cell interactions in culture as a first step toward potentially using such polymers to stimulate in vivo nerve regeneration. Image analysis demonstrates that PC-12 cells and primary chicken sciatic nerve explants attached and extended neurites equally well on both PP films and tissue culture polystyrene in the absence of electrical stimulation. In contrast, PC-12 cells interacted poorly with indium tin oxide (ITO), poly(l-lactic acid) (PLA), and poly(lactic acid-co-glycolic acid) surfaces. However, PC-12 cells cultured on PP films and subjected to an electrical stimulus through the film showed a significant increase in neurite lengths compared with ones that were not subjected to electrical stimulation through the film and tissue culture polystyrene controls. The median neurite length for PC-12 cells grown on PP and subjected to an electrical stimulus was 18.14 μm (n = 5643) compared with 9.5 μm (n = 4440) for controls. Furthermore, animal implantation studies reveal that PP invokes little adverse tissue response compared with poly(lactic acid-co-glycolic acid).

Absence of a barrier to backwards rotation of the bacterial flagellar motor demonstrated with optical tweezers

A cell of the bacterium Escherichia coli was tethered covalently to a glass coverslip by a single flagellum, and its rotation was stopped by using optical tweezers. The tweezers acted directly on the cell body or indirectly, via a trapped polystyrene bead. The torque generated by the flagellar motor was determined by measuring the displacement of the laser beam on a quadrant photodiode. The coverslip was mounted on a computer-controlled piezo-electric stage that moved the tether point in a circle around the center of the trap so that the speed of rotation of the motor could be varied. The motor generated ≈4500 pN nm of torque at all angles, regardless of whether it was stalled, allowed to rotate very slowly forwards, or driven very slowly backwards. This argues against models of motor function in which rotation is tightly coupled to proton transit and back-transport of protons is severely limited.

Modulation of Cell Proliferation and Differentiation through Substrate-dependent Changes in Fibronectin Conformation

Integrin-mediated cell adhesion to extracellular matrices provides signals essential for cell cycle progression and differentiation. We demonstrate that substrate-dependent changes in the conformation of adsorbed fibronectin (Fn) modulated integrin binding and controlled switching between proliferation and differentiation. Adsorption of Fn onto bacterial polystyrene (B), tissue culture polystyrene (T), and collagen (C) resulted in differences in Fn conformation as indicated by antibody binding. Using a biochemical method to quantify bound integrins in cultured cells, we found that differences in Fn conformation altered the quantity of bound α5 and β1 integrin subunits but not αv or β3. C2C12 myoblasts grown on these Fn-coated substrates proliferated to different levels (B > T > C). Immunostaining for muscle-specific myosin revealed minimal differentiation on B, significant levels on T, and extensive differentiation on C. Differentiation required binding to the RGD cell binding site in Fn and was blocked by antibodies specific for this site. Switching between proliferation and differentiation was controlled by the levels of α5β1 integrin bound to Fn, and differentiation was inhibited by anti-α5, but not anti-αv, antibodies, suggesting distinct integrin-mediated signaling pathways. Control of cell proliferation and differentiation through conformational changes in extracellular matrix proteins represents a versatile mechanism to elicit specific cellular responses for biological and biotechnological applications.