Institutionalising New Forms of Environmental Policy-making in the EU. The Case of the Emissions Trading Scheme.

The Emissions Trading Scheme represents a new departure in the field of environmental policy-making in the EU. With it, the EU has deployed a market-based instrument to meet its Kyoto Carbon Dioxide emissions reduction commitments. This paper will account for its emergence as a concrete policy output. It will first refer to the historical, ideational and interest-based logics which shaped its emergence, within the broader context of environmental policy-making in the EU. Using an integrated institutionalist approach incorporating all three strands of institutionalist thought, it will therefore account for the sequencing of these different logics at different stages of the policy-making process, from agenda-setting through to implementation and evaluation. Accordingly, it will be argued that this approach adds value in accounting for the change in logics which can often lead from good ideas unintentionally emerging as sub-optimal policies in the EU, as is the case with the first phase of the Emissions Trading Scheme

Spatiotemporal organisation of protein nanoclusters in adhesion complexes

The main goal of this thesis was to contribute to the understanding of the nanoscale lateral organisation of key proteins in adhesion complexes. For this, we exploited single molecule localisation-based super-resolution microscopy STORM to visualise the lateral organisation of five key proteins of the adhesion complex: the integrins, a5ß1 and avß3, and three of their adaptor proteins: paxillin, talin, and vinculin. We first established that these proteins form nanoclusters of around 50nm size that are preserved across all five proteins. Interestingly, these nanoclusters have similar size and number of localisations regardless of their localisation on the membrane, i.e., in the different adhesion structures studied, namely, FA and fAs as well as outside, and were maintained for different cell seeding times, from 90 min to 24 h. These results suggest that nanoclustering constitutes a general mechanism of adhesion protein organisation, creating nanohubs of functional activity. When studying how protein organisation in nanoclusters changes as a function of adhesion time, we revealed a two- and a four-fold increase in the density of a5ß1 and avß3 clusters, respectively, for cells that spread for 24 h as compared to those that spread for 90 min. Further analysis suggests that the increase in density of integrin nanoclusters is due to selective targeting of new integrin nanoclusters to the basal membrane. Following on from this, we then focus on mapping the distribution of these nanoclusters, first by measuring the nearest neighbour distance; (NND) between clusters of the same protein, and second by considering the shortest distance between clusters of different proteins. We found a clear physical segregation of nanoclusters of the same protein around ~55 nm, which is established at early time points after cell seeding for a5ß1 and the adaptors and maintained after 24 h. Interestingly, avß3 nanoclusters exhibited a more random distribution at earlier seeding times and progressively reached similar lateral segregation at 24 h. Concomitant with this lateral segregation, we observed an enriched of all proteins at distances between 100-200 nm. Our observations are in line with the existence of a critical distance spacing between integrins needed for support adhesion and stabilisation of focal adhesions. Furthermore, we found that the relative distribution of nanoclusters of different proteins is predominantly random, with the exception of a5ß1 and paxillin, which organise with a separation of 50 nm. Such an unexpected random distribution between integrins and their adaptors might reflect the dynamic and short-live active state of integrins. Finally, we evaluated and described the mesoscale organisation of nanoclusters inside adhesions. Specifically, we computed the shortest distance between a nanocluster and the edge of the adhesion and studied how the distance to the edge depends on the NND between clusters of different proteins. Remarkably, we found a preference for a5ß1 nanoclusters to be at the edge of the adhesions and in close proximity to its adaptors in a peripheral belt region of the adhesions. Altogether, the results of this thesis demonstrate a clear lateral and hierarchical organisation of integrins and their adaptors inside focal adhesions. Based on our results (together with extensive literature in the field), we propose that one population of a5ß1 nanoclusters and their adaptors preferentially localise close to the edge of adhesion complexes regulating the process of adhesion. A second population of a5ß1 and most of the avß3 nanoclusters organise more randomly at the centre of the adhesions, with dynamic and brief engagement to their adaptors, likely playing a role in mechanotransduction. As a whole, we postulate that the lateral nano- and meso-scale organisation of adhesion proteins is strictly related to and important for the functions of adhesion, mechanosensing and mechanotransduction.El objetivo de esta tesis ha sido contribuir a la comprensión de la organización lateral a nanoescala de proteínas clave en complejos de adhesión. Para ello, usamos la microscopía de superresolución STORM, para visualizar con resolución espacial nanométrica la organización lateral de cinco proteínas del complejo de adhesión: dos integrinas, a5ß1 y avß3, y las proteínas adaptadoras: paxilin, talin y vinculin. En primer lugar, establecimos que estas proteínas forman nanoagregados de ~50 nm tamaño en las cinco proteínas. Curiosamente, su tamaño y número de localizaciones son similares, independientemente de su localización en la membrana, es decir, tanto en FA y fAs, así como fuera de las adhesiones, manteniéndose constantes durante diferentes tiempos de siembra celular. Estos resultados sugieren que la nanoagregación constituye un mecanismo general de organización de proteínas de adhesión, constituyendo nanocentros de actividad funcional. Además, revelamos un aumento en la densidad de los agregados de a5ß1 y avß3 en células extendidas por 24 h en comparación con 90 min, mientras que la densidad de agregados de las proteínas adaptadoras se mantuvo constante. Esta disparidad en densidades indica que solo una fracción de las integrinas interacciona con sus adaptadores, consistente con estados dinámicos de activación-desactivación de las integrinas. También nos enfocamos en mapear la distribución de estos nanoagregados, midiendo la distancia entre agregados más corta entre grupos de la misma proteína, y luego, considerando la distancia más corta entre grupos de diferentes proteínas. Encontramos una clara segregación física de agregados de la misma proteína alrededor de ~55 nm, que se establece temprano después de la siembra celular para a5ß1 y sus adaptadores, y se mantiene hasta 24 h. Curiosamente, los agregados de avß3 exhibieron una distribución más aleatoria en tiempos tempranos de siembra, alcanzando progresivamente una segregación lateral similar a 24 h. Acompañada a esta segregación lateral, observamos un enriquecimiento de todas las proteínas a distancias entre 100¿200 nm. Nuestras observaciones son consistentes con la existencia de un espaciado de distancia crítico entre las integrinas necesarias para apoyar la adhesión y estabilizar las adhesiones focales. Además, encontramos que la distribución relativa de nanoagregados de diferentes proteínas es aleatoria, lo cual podría reflejar el estado activo dinámico y de corta duración de las integrinas, de modo que con nuestras condiciones de imágenes, actualmente no podemos capturar la participación de aquellas integrinas activas dentro de la población total. Finalmente, evaluamos la organización de mesoescala de nanoagregados en FAs, específicamente, en los bordes y el centro. Sorprendentemente, encontramos una preferencia por nanoagregados de a5ß1 en el borde de las FAs y cerca de sus adaptadores, en una región periférica a los bordes. En conjunto, nuestros resultados demuestran una clara organización lateral y jerárquica de las integrinas y sus adaptadores dentro de las adhesiones focales. Proponemos que una población de nanoagregados de a5ß1 y sus adaptadores se localizan preferentemente cerca del borde de los complejos de adhesión para regular el proceso de adhesión y probablemente interaccionando activamente con la maquinaria de la actomiosina. Una segunda población de a5ß1 y la mayoría de los nano-gregados de avß3 se organizan de forma aleatoria en el centro de las FAs con una interacción dinámica y breve con sus adaptadores, posiblemente comprometidos con el proceso de mecanotransducción. En conjunto, y similar a su organización axial, postulamos que la organización lateral a nano- y meso-escala dentro de las FAs es importante para las funciones de adhesión, mecanosensibilidad y mecanotransducción.Postprint (published version

Affective Computing for Emotion Detection using Vision and Wearable Sensors

The research explores the opportunities, challenges, limitations, and presents advancements in computing that relates to, arises from, or deliberately influences emotions (Picard, 1997). The field is referred to as Affective Computing (AC) and is expected to play a major role in the engineering and development of computationally and cognitively intelligent systems, processors and applications in the future. Today the field of AC is bolstered by the emergence of multiple sources of affective data and is fuelled on by developments under various Internet of Things (IoTs) projects and the fusion potential of multiple sensory affective data streams. The core focus of this thesis involves investigation into whether the sensitivity and specificity (predictive performance) of AC, based on the fusion of multi-sensor data streams, is fit for purpose? Can such AC powered technologies and techniques truly deliver increasingly accurate emotion predictions of subjects in the real world? The thesis begins by presenting a number of research justifications and AC research questions that are used to formulate the original thesis hypothesis and thesis objectives. As part of the research conducted, a detailed state of the art investigations explored many aspects of AC from both a scientific and technological perspective. The complexity of AC as a multi-sensor, multi-modality, data fusion problem unfolded during the state of the art research and this ultimately led to novel thinking and origination in the form of the creation of an AC conceptualised architecture that will act as a practical and theoretical foundation for the engineering of future AC platforms and solutions. The AC conceptual architecture developed as a result of this research, was applied to the engineering of a series of software artifacts that were combined to create a prototypical AC multi-sensor platform known as the Emotion Fusion Server (EFS) to be used in the thesis hypothesis AC experimentation phases of the research. The thesis research used the EFS platform to conduct a detailed series of AC experiments to investigate if the fusion of multiple sensory sources of affective data from sensory devices can significantly increase the accuracy of emotion prediction by computationally intelligent means. The research involved conducting numerous controlled experiments along with the statistical analysis of the performance of sensors for the purposes of AC, the findings of which serve to assess the feasibility of AC in various domains and points to future directions for the AC field. The AC experiments data investigations conducted in relation to the thesis hypothesis used applied statistical methods and techniques, and the results, analytics and evaluations are presented throughout the two thesis research volumes. The thesis concludes by providing a detailed set of formal findings, conclusions and decisions in relation to the overarching research hypothesis on the sensitivity and specificity of the fusion of vision and wearables sensor modalities and offers foresights and guidance into the many problems, challenges and projections for the AC field into the future

Editorial: Ethics in TESOL

This special issue of TESOL in Context focuses on the topic of “Ethics in TESOL”. In it a number of issues are explored including accountability of ESOL teachers to their students and communities, the development of ethical responsibility for professional life and how research can inform and transform practice. Broader questions are raised about where does the notion of ethics take us as a profession in relation to research, practice and advocacy