A study on X-ray irradiation of composite polyaniline LB films

Composite Langmuir-Blodgett (LB) films from polyaniline and cadmium stearate have been irradiated with ionizing X-rays for various exposure times. In the initial stages of X-ray irradiation the absorption peak at 580 nm of an as-deposited film was seen to decrease with a concomitant increase in the absorption in the long wavelength region (700-1100 nm). Upon prolonging the irradiation, the absorption maximum shifted to 800 nm with the LB film color changing to green, characteristic of acid doped polyaniline. The changes in the Fourier transform infrared (FTIR) spectra upon irradiation are also similar to those observed upon acid doping of polyaniline. When compared with acid doping, two major differences were observed for the LB films exposed to X-rays. First, the packing order of the cadmium stearate domains in the composite LB films - as observed by X-ray diffraction - is not affected by the X-ray irradiation. In addition, no significant increase in the DC conductivity was noted after the X-ray exposure whereas similar LB films have their conductivity increased by an order of magnitude upon acid doping. These differences may be explained by considering that the inter-domain contribution to the conductivity is increased by the acid doping because the insulating cadmium stearate domains are destroyed, which does not occur with the X-ray irradiation. (C) 1998 Elsevier B.V. S.A. All rights reserved.

Contribution towards intelligent service management in wearable and ubiquitous devices

Hoy en día asistimos a un creciente interés por parte de la sociedad hacia el cuidado de la salud. Esta afirmación viene apoyada por dos realidades. Por una parte, el aumento de las prácticas saludables (actividad deportiva, cuidado de la alimentación, etc.). De igual manera, el auge de los dispositivos inteligentes (relojes, móviles o pulseras) capaces de medir distintos parámetros físicos como el pulso cardíaco, el ritmo respiratorio, la distancia recorrida, las calorías consumidas, etc. Combinando ambos factores (interés por el estado de salud y disponibilidad comercial de dispositivos inteligentes) están surgiendo multitud de aplicaciones capaces no solo de controlar el estado actual de salud, también de recomendar al usuario cambios de hábitos que lleven hacia una mejora en su condición física. En este contexto, los llamados dispositivos llevables (weareables) unidos al paradigma de Internet de las cosas (IoT, del inglés Internet of Things) permiten la aparición de nuevos nichos de mercado para aplicaciones que no solo se centran en la mejora de la condición física, ya que van más allá proponiendo soluciones para el cuidado de pacientes enfermos, la vigilancia de niños o ancianos, la defensa y la seguridad, la monitorización de agentes de riesgo (como bomberos o policías) y un largo etcétera de aplicaciones por llegar. El paradigma de IoT se puede desarrollar basándose en las existentes redes de sensores inalámbricos (WSN, del inglés Wireless Sensor Network). La conexión de los ya mencionados dispositivos llevables a estas redes puede facilitar la transición de nuevos usuarios hacia aplicaciones IoT. Pero uno de los problemas intrínsecos a estas redes es su heterogeneidad. En efecto, existen multitud de sistemas operativos, protocolos de comunicación, plataformas de desarrollo, soluciones propietarias, etc. El principal objetivo de esta tesis es realizar aportaciones significativas para solucionar no solo el problema de la heterogeneidad, sino también de dotar de mecanismos de seguridad suficientes para salvaguardad la integridad de los datos intercambiados en este tipo de aplicaciones. Algo de suma importancia ya que los datos médicos y biométricos de los usuarios están protegidos por leyes nacionales y comunitarias. Para lograr dichos objetivos, se comenzó con la realización de un completo estudio del estado del arte en tecnologías relacionadas con el marco de investigación (plataformas y estándares para WSNs e IoT, plataformas de implementación distribuidas, dispositivos llevables y sistemas operativos y lenguajes de programación). Este estudio sirvió para tomar decisiones de diseño fundamentadas en las tres contribuciones principales de esta tesis: un bus de servicios para dispositivos llevables (WDSB, Wearable Device Service Bus) basado en tecnologías ya existentes tales como ESB, WWBAN, WSN e IoT); un protocolo de comunicaciones inter-dominio para dispositivos llevables (WIDP, Wearable Inter-Domain communication Protocol) que integra en una misma solución protocolos capaces de ser implementados en dispositivos de bajas capacidades (como lo son los dispositivos llevables y los que forman parte de WSNs); y finalmente, la tercera contribución relevante es una propuesta de seguridad para WSN basada en la aplicación de dominios de confianza. Aunque las contribuciones aquí recogidas son de aplicación genérica, para su validación se utilizó un escenario concreto de aplicación: una solución para control de parámetros físicos en entornos deportivos, desarrollada dentro del proyecto europeo de investigación “LifeWear”. En este escenario se desplegaron todos los elementos necesarios para validar las contribuciones principales de esta tesis y, además, se realizó una aplicación para dispositivos móviles por parte de uno de los socios del proyecto (lo que contribuyó con una validación externa de la solución). En este escenario se usaron dispositivos llevables tales como un reloj inteligente, un teléfono móvil con sistema operativo Android y un medidor del ritmo cardíaco inalámbrico capaz de obtener distintos parámetros fisiológicos del deportista. Sobre este escenario se realizaron diversas pruebas de validación mediante las cuales se obtuvieron resultados satisfactorios. ABSTRACT Nowadays, society is shifting towards a growing interest and concern on health care. This phenomenon can be acknowledged by two facts: first, the increasing number of people practising some kind of healthy activity (sports, balanced diet, etc.). Secondly, the growing number of commercial wearable smart devices (smartwatches or bands) able to measure physiological parameters such as heart rate, breathing rate, distance or consumed calories. A large number of applications combining both facts are appearing. These applications are not only able to monitor the health status of the user, but also to provide recommendations about routines in order to improve the mentioned health status. In this context, wearable devices merged with the Internet of Things (IoT) paradigm enable the proliferation of new market segments for these health wearablebased applications. Furthermore, these applications can provide solutions for the elderly or baby care, in-hospital or in-home patient monitoring, security and defence fields or an unforeseen number of future applications. The introduced IoT paradigm can be developed with the usage of existing Wireless Sensor Networks (WSNs) by connecting the novel wearable devices to them. In this way, the migration of new users and actors to the IoT environment will be eased. However, a major issue appears in this environment: heterogeneity. In fact, there is a large number of operating systems, hardware platforms, communication and application protocols or programming languages, each of them with unique features. The main objective of this thesis is defining and implementing a solution for the intelligent service management in wearable and ubiquitous devices so as to solve the heterogeneity issues that are presented when dealing with interoperability and interconnectivity of devices and software of different nature. Additionally, a security schema based on trust domains is proposed as a solution to the privacy problems arising when private data (e.g., biomedical parameters or user identification) is broadcasted in a wireless network. The proposal has been made after a comprehensive state-of-the-art analysis, and includes the design of a Wearable Device Service Bus (WDSB) including the technologies collected in the requirement analysis (ESB, WWBAN, WSN and IoT). Applications are able to access the WSN services regardless of the platform and operating system where they are running. Besides, this proposal also includes the design of a Wearable Inter-Domain communication Protocols set (WIDP) which integrates lightweight protocols suitable to be used in low-capacities devices (REST, JSON, AMQP, CoAP, etc...). Furthermore, a security solution for service management based on a trustworthy domains model to deploy security services in WSNs has been designed. Although the proposal is a generic framework for applications based on services provided by wearable devices, an application scenario for testing purposes has been included. In this validation scenario it has been presented an autonomous physical condition performance system, based on a WSN, bringing the possibility to include several elements in an IoT scenario: a smartwatch, a physiological monitoring device and a smartphone. In summary, the general objective of this thesis is solving the heterogeneity and security challenges arising when developing applications for WSNs and wearable devices. As it has been presented in the thesis, the solution proposed has been successfully validated in a real scenario and the obtained results were satisfactory.

Structural basis for heterogeneous kinetics: Reengineering the hairpin ribozyme

The RNA cleavage reaction catalyzed by the hairpin ribozyme shows biphasic kinetics, and chase experiments show that the slow phase of the reaction results from reversible substrate binding to an inactive conformational isomer. To investigate the structural basis for the heterogeneous kinetics, we have developed an enzymatic RNA modification method that selectively traps substrate bound to the inactive conformer and allows the two forms of the ribozyme-substrate complex to be separated and analyzed by using both physical and kinetic strategies. The inactive form of the complex was trapped by the addition of T4 RNA ligase to a cleavage reaction, resulting in covalent linkage of the 5′ end of the substrate to the 3′ end of the ribozyme and in selective and quantitative ablation of the slow kinetic phase of the reaction. This result indicates that the inactive form of the ribozyme-substrate complex can adopt a conformation in which helices 2 and 3 are coaxially stacked, whereas the active form does not have access to this conformation, because of a sharp bend at the helical junction that presumably is stabilized by inter-domain tertiary contacts required for catalytic activity. These results were used to improve the activity of the hairpin ribozyme by designing new interfaces between the two domains, one containing a non-nucleotidic orthobenzene linkage and the other replacing the two-way junction with a three-way junction. Each of these modified ribozymes preferentially adopts the active conformation and displays improved catalytic efficiency.

The (Un)Folding of Multidomain Proteins Through the Lens of Single-molecule Force-spectroscopy and Computer Simulation

Proteins are specialized molecules that catalyze most of the reactions that can sustain life, and they become functional by folding into a specific 3D structure. Despite their importance, the question, "how do proteins fold?" - first pondered in in the 1930's - is still listed as one of the top unanswered scientific questions as of 2005, according to the journal Science. Answering this question would provide a foundation for understanding protein function and would enable improved drug targeting, efficient biofuel production, and stronger biomaterials. Much of what we currently know about protein folding comes from studies on small, single-domain proteins, which may be quite different from the folding of large, multidomain proteins that predominate the proteomes of all organisms.

In this thesis I will discuss my work to fill this gap in understanding by studying the unfolding and refolding of large, multidomain proteins using the powerful combination of single-molecule force-spectroscopy experiments and molecular dynamic simulations.

The three model proteins studied - Luciferase, Protein S, and Streptavidin - lend insight into the inter-domain dependence for unfolding and the subdomain stabilization of binding ligands, and ultimately provide new insight into atomistic details of the intermediate states along the folding pathway.

Inter-observer agreement for spectral- and time-domain optical coherence tomography image grading: a prospective study

The purpose of this study was to compare inter-observer agreement of Stratus™ OCT versus Spectralis™ OCT image grading in patients with neovascular age-related macular degeneration (AMD). Thirty eyes with neovascular AMD were examined with Stratus™ OCT and Spectralis™ OCT. Four different scan protocols were used for imaging. Three observers graded the images for the presence of various pathologies. Inter-observer agreement between OCT models was assessed by calculating intra-class correlation coefficients (ICC). In Stratus™ OCT highest interobserver agreement was found for subretinal fluid (ICC: 0.79), and in Spectralis™ OCT for intraretinal cysts (IRC) (ICC: 0.93). Spectralis™ OCT showed superior interobserver agreement for IRC and epiretinal membranes (ERM) (ICC(Stratus™): for IRC 0.61; for ERM 0.56; ICC(Spectralis™): for IRC 0.93; for ERM 0.84). Increased image resolution of Spectralis™ OCT did improve the inter-observer agreement for grading intraretinal cysts and epiretinal membranes but not for other retinal changes.

Eukaryotic translation elongation factor 1A (eEF1A) domain I from S. cerevisiae is required but not sufficient for inter-species complementation

Ethanolamine phosphoglycerol (EPG) is a protein modification attached exclusively to eukaryotic elongation factor 1A (eEF1A). In mammals and plants, EPG is linked to conserved glutamate residues located in eEF1A domains II and III, whereas in the unicellular eukaryote Trypanosoma brucei, only domain III is modified by a single EPG. A biosynthetic precursor of EPG and structural requirements for EPG attachment to T. brucei eEF1A have been reported, but nothing is known about the EPG modifying enzyme(s). By expressing human eEF1A in T. brucei, we now show that EPG attachment to eEF1A is evolutionarily conserved between T. brucei and Homo sapiens. In contrast, S. cerevisiae eEF1A, which has been shown to lack EPG is not modified in T. brucei. Furthermore, we show that eEF1A cannot functionally complement across species when using T. brucei and S. cerevisiae as model organisms. However, functional complementation in yeast can be obtained using eEF1A chimera containing domains II or III from other species. In contrast, yeast domain I is strictly required for functional complementation in S. cerevisiae.

Regulation of Nedd4-2 self-ubiquitination and stability by a PY motif located within its HECT-domain.

Ubiquitin ligases play a pivotal role in substrate recognition and ubiquitin transfer, yet little is known about the regulation of their catalytic activity. Nedd4 (neural-precursor-cell-expressed, developmentally down-regulated 4)-2 is an E3 ubiquitin ligase composed of a C2 domain, four WW domains (protein-protein interaction domains containing two conserved tryptophan residues) that bind PY motifs (L/PPXY) and a ubiquitin ligase HECT (homologous with E6-associated protein C-terminus) domain. In the present paper we show that the WW domains of Nedd4-2 bind (weakly) to a PY motif (LPXY) located within its own HECT domain and inhibit auto-ubiquitination. Pulse-chase experiments demonstrated that mutation of the HECT PY-motif decreases the stability of Nedd4-2, suggesting that it is involved in stabilization of this E3 ligase. Interestingly, the HECT PY-motif mutation does not affect ubiquitination or down-regulation of a known Nedd4-2 substrate, ENaC (epithelial sodium channel). ENaC ubiquitination, in turn, appears to promote Nedd4-2 self-ubiquitination. These results support a model in which the inter- or intra-molecular WW-domain-HECT PY-motif interaction stabilizes Nedd4-2 by preventing self-ubiquitination. Substrate binding disrupts this interaction, allowing self-ubiquitination of Nedd4-2 and subsequent degradation, resulting in down-regulation of Nedd4-2 once it has ubiquitinated its target. These findings also point to a novel mechanism employed by a ubiquitin ligase to regulate itself differentially compared with substrate ubiquitination and stability.

1.° Bartholomaei Facii ad Alphonsum Regem dialogus de vitae felicitate. — 2.° Ejusdem epistola ad Robertum Strozam. — 3.° Mafei Vegii, Laudensis, dialogus de felicitate et miseria. — 4.° Ejusdem ad Eustachium, fratrem suum, dialogus de injuriis veritati illatis. — 5.° Anonymi dialogus inter Epicureum et Stoïcum de virtute et voluptate. — 6.° Platonis dialogus de morte contemnenda : interprete Cincio, Romano. — 7.° Plutarchi sermo de virtute et vitio : eodem interprete. — 8.° Apologia Socratis : authore Platone ; interprete verò Leonardo Aretino. — 9.° Platonis Crito : interprete anonymo. — 10.° Luciani Toxaris, sive, dialogus de amicitia : interprete anonymo.

Rogerii de Gaignieres

1.° M. T. Ciceronis ad Caium Trebatium, Jurisconsultum, topica : passim inter lineas glossae. — 2.° A. Manlii Severini Boëtii in Ciceronis topica commentariorum libri sex.

Colbertinus

1.° M. T. Ciceronis rhetoricorum ad Herennium libri quatuor. — 2.° Joannis Boni Andreae, civis Bononiensis, ars dictaminis. — 3.° Joannis Candidi, Secretarii Imperialis, liber artis novae epistolarum. — 4.° Forma appellationis Raymundi de Oppeda, Sistarciensis Episcopi. — 5.° Ars memoriae secundùm Aristotelem et S. Thomam de Aquino. — 6.° Tractatus de baculo Jacob : authore anonymo. — 7.° Exordia secundùm Cassiodorum, de pace facienda inter Imperatorem et Regem. — 8.° Petri Blesensis epistolae nonnullae. — 9.° Guidonis de Columna historiae Trojanae libri decem priores.

Colbertinus

Gualteri Castellionensis Alexandreis : passim inter lineas glossae et ad marginem scholia.

Colbertinus

Quinti Horatii Flacci opera omnia : passim inter lineas glossae et ad marginem scholia ; ad calcem subjiciuntur anonymi annotationes de numeris Horatii, et ejusdem vita.

Colbertinus

P. Ovidii Nasonis metamorphoseon libri quindecim : passim inter lineas glossae et ad marginem scholia.

Colbertinus, antea Jacobi Augusti Thuani