Lfc is a guanine nucleotide exchange factor (GEF) that activates the small GTPase RhoA, and its GEF activity is tightly regulated through protein-protein interactions, phosphorylation, and cellular localization. Lfc is anchored to microtubules through its interaction with the dynein light chain Tctex-1, which results in inhibition of Lfc's GEF activity. Here we present a crystallographic structure of Tctex-1 in complex with Lfc with residues 143-155 of Lfc bound at the Tctex-1 dimer interface. Structural alignment of our structure with Tctex-1 in complex with the dynein intermediate chain (DIC) shows the binding site of the DIC peptide and Lfc substantially overlap. Biochemical evidence, NMR perturbations assays and intrinsic fluorescence provide structural validation and support an extension of the Lfc binding site to the α-helices that may accommodate additional contact points with Tctex-1. We postulate a potential mechanism for Lfc’s recruitment to the microtubules through a tripartite complex with Tctex-1 and DIC.

Lfc is a guanine nucleotide exchange factor (GEF) that activates the small GTPase RhoA, and its GEF activity is tightly regulated through protein-protein interactions, phosphorylation, and cellular localization. Lfc is anchored to microtubules through its interaction with the dynein light chain Tctex-1, which results in inhibition of Lfc's GEF activity. Here we present a crystallographic structure of Tctex-1 in complex with Lfc with residues 143-155 of Lfc bound at the Tctex-1 dimer interface. Structural alignment of our structure with Tctex-1 in complex with the dynein intermediate chain (DIC) shows the binding site of the DIC peptide and Lfc substantially overlap. Biochemical evidence, NMR perturbations assays and intrinsic fluorescence provide structural validation and support an extension of the Lfc binding site to the α-helices that may accommodate additional contact points with Tctex-1. We postulate a potential mechanism for Lfc’s recruitment to the microtubules through a tripartite complex with Tctex-1 and DIC.

Tese de mestrado integrado. Engenharia Civil. Faculdade de Engenharia. Universidade do Porto. 2010

During development of the mammalian nervous system, neural stem cells generate neurons first and glia second, thereby allowing the initial establishment of neuronal circuitry, and subsequent matching of glial numbers and position to that circuitry. Multiple molecular mechanisms act in concert to control neural precursor expansion prior to neurogenesis, and to allow for an exponential generation of neurons while ensuring the maintenance of sufficient precursors to produce later-born neurons, glial cells and adult neural stem cells. Throughout cortical development, these processes are regulated in part by the precursors’ environment as well as intrinsic changes in precursors and their modes of division, which regulate the fate of daughter cells and the balance between self-renewal and differentiation. In the first part of this thesis, the protein tyrosine phosphatase SHP-2 was identified as a novel signaling protein that regulates the neurogenic to gliogenic switch by potentiating neurogenic signals and suppressing gliogenic signals until the appropriate developmental time point for astrogenesis, providing one mechanism whereby precursors integrate conflicting environmental cues. A Noonan Syndrome (NS)-associated activated SHP-2 mutation causes perturbations in neural cell genesis, which may contribute to the mild mental retardation and learning disabilities observed in NS patients. In the second part of this thesis, a novel Rho-regulatory pathway which includes the Rho-GEF Lfc and its negative regulator Tctex-1 were also found to regulate neurogenesis, potentially by directing mitotic spindle orientation during precursor divisions, thereby regulating the symmetric and asymmetric nature of radial precursor divisions.

This thesis proposes and demonstrates a hydrogenated amorphous silicon passivated, inverted photovoltaic device on n-type silicon, utilizing a Laser Fired Emitter on a rear i-a- Si:H/SiOx dielectric stack. This novel low-temperature-fabricated device architecture constitutes the first demonstration of an LFE on a dielectric passivation stack. The optimization of the device is explored through Sentaurus computational modeling, predicting a potential efficiency of >20%. Proof of concept devices are fabricated using the DC Saddle Field PECVD system for the deposition of hydrogenated amorphous silicon passivation layers. Laser parameters are explored highlighting pulse energy density as a key performance determining factor. Annealing of devices in nitrogen atmosphere shows performance improvements albeit that the maximum annealing temperature is limited by the thermal stability of the passivation. A proof of concept device efficiency of 11.1% is realized forming the basis for further device optimization.

This thesis proposes and demonstrates a hydrogenated amorphous silicon passivated, inverted photovoltaic device on n-type silicon, utilizing a Laser Fired Emitter on a rear i-a- Si:H/SiOx dielectric stack. This novel low-temperature-fabricated device architecture constitutes the first demonstration of an LFE on a dielectric passivation stack. The optimization of the device is explored through Sentaurus computational modeling, predicting a potential efficiency of >20%. Proof of concept devices are fabricated using the DC Saddle Field PECVD system for the deposition of hydrogenated amorphous silicon passivation layers. Laser parameters are explored highlighting pulse energy density as a key performance determining factor. Annealing of devices in nitrogen atmosphere shows performance improvements albeit that the maximum annealing temperature is limited by the thermal stability of the passivation. A proof of concept device efficiency of 11.1% is realized forming the basis for further device optimization.

During development of the mammalian nervous system, neural stem cells generate neurons first and glia second, thereby allowing the initial establishment of neuronal circuitry, and subsequent matching of glial numbers and position to that circuitry. Multiple molecular mechanisms act in concert to control neural precursor expansion prior to neurogenesis, and to allow for an exponential generation of neurons while ensuring the maintenance of sufficient precursors to produce later-born neurons, glial cells and adult neural stem cells. Throughout cortical development, these processes are regulated in part by the precursors’ environment as well as intrinsic changes in precursors and their modes of division, which regulate the fate of daughter cells and the balance between self-renewal and differentiation. In the first part of this thesis, the protein tyrosine phosphatase SHP-2 was identified as a novel signaling protein that regulates the neurogenic to gliogenic switch by potentiating neurogenic signals and suppressing gliogenic signals until the appropriate developmental time point for astrogenesis, providing one mechanism whereby precursors integrate conflicting environmental cues. A Noonan Syndrome (NS)-associated activated SHP-2 mutation causes perturbations in neural cell genesis, which may contribute to the mild mental retardation and learning disabilities observed in NS patients. In the second part of this thesis, a novel Rho-regulatory pathway which includes the Rho-GEF Lfc and its negative regulator Tctex-1 were also found to regulate neurogenesis, potentially by directing mitotic spindle orientation during precursor divisions, thereby regulating the symmetric and asymmetric nature of radial precursor divisions.

This study aims to explore teacher beliefs on accents in the classroom and how they affect classroom practices as well as the teachers views on the LFC approach. Furthermore, the study investigates whether there exists a possible disconnect between teachers and the Swedish National Agency for Education when it comes to deciding what approach to take when teaching English pronunciation. The study is based on semi-structured interviews with five Secondary school and Upper Secondary School English teachers in Sweden, whose work experience ranged from six months to 22 years. The study found that the teachers with more experience were more likely to prefer the inner circle English accents and use them as a benchmark for correctness. Phenomena like the native-speaker ideal, which means to idealize native speakers and view them as better speakers of English, can be linked to the teachers age and experience. Moreover, some teachers expressed feeling pressure from students to sound native-like and felt disfavored as some schools showed a preference for hiring native speakers as English teachers. Thus, a haloeffect for teachers speaking with an inner-circle English accent was identified, which affects students’ and employers’ perception of non-native EFL-teachers. The study also found that while some of the teachers were familiar with the Lingua Franca Core model, none of them had taught it. The study concludes that there is a disconnect between the teachers and Skolverket’s steering documents, as most of the teachers felt that Skolverket does not explicitly call for pronunciation teaching and were unsure of the demands. Furthermore, this calls for clearer demands from Skolverket as well as re-formation programs for experienced teacher to change their beliefs on non-native accents.

En France, les chaudières à lit fluidisé circulant (LFC) ont une vingtaine d'années d'existence. Malgré leur âge, l'intérêt de leur système de désulfuration des fumées est amoindri du fait de la difficulté à valoriser les cendres qu'elles produisent. A l'heure actuelle, les voies de valorisation envisageables pour ces co-produits sont faiblement consommatrices de cendre et la filière du génie civil, c'est-à-dire la confection de ciment et de béton, leur reste inaccessible. Ces travaux de thèse visent à démontrer que sous réserves de plusieurs précautions, il est possible et même intéressant de confectionner un ciment contenant la cendre volante LFC. Du point de vue minéralogique, les cendres volantes LFC se différencient clairement des cendres volantes silico-alumineuses issues des chaudières à charbon pulvérisé (CP) normalisées par la présence de minéraux argileux, proches de ceux contenus dans les combustibles. A l'aide de la diffraction des rayons X et d'un logiciel de traitement de données par méthode de Rietveld, plusieurs couples de combustible et de cendre ont été caractérisés afin d'étudier leur filiation minéralogique. Du point de vue chimique, le système de désulfuration des chaudières LFC conduit à une grande diversité de composition chimique, de par l'ajout en quantité variable d'une fraction sulfo-calcique aux résidus silico-alumineux de la combustion des charbons. Cette fraction, constituée d'anhydrite et de chaux vive, est le principal frein à la valorisation de la cendre dans la filière du génie civil. Que la cendre soit hydratée seule ou en mélange avec du ciment, son anhydrite conduit à la formation d'ettringite, hydrate responsable de désordres dans la pâte de ciment. Les travaux de thèse eurent comme objectif d'étudier ce phénomène, en tâchant de le relier aux variations dimensionnelles de mortier et de béton. Au final, nos résultats dressent une liste de spécifications, suffisantes pour rendre réalisable la confection d'un ciment composé destiné à une utilisation industrielle. De plus, des essais mécaniques montrent que la nature physique, chimique et minéralogique de la cendre constitue un avantage en terme de résistance mécanique pour confectionner des ciments composés. En détaillant les principaux facteurs qui influencent le gonflement ettringitique et en apportant un nouvel éclairage sur la nature minéralogique de la cendre, cette étude pourrait aider, à terme, à la prise en compte de cette addition minérale dans les normes relatives à la confection de ciment et de béton.

This paper aims at investigating how upper secondary school teachers of English in Malmö, Sweden abide by the curriculum criteria of having their learners develop correctness in speech, as well as what support English teachers receive with regard to pronunciation teaching from official steering documents. Furthermore, this paper attempts to critically evaluate and discuss potential options for pronunciation models or standards in education which hold sway in contemporary research. Through qualitative interviews with four upper secondary school teachers of English, the findings of this study indicate a discrepancy between the participants’ views on how ‘correctness’ should be interpreted as well as their methods for teaching and assessing pronunciation. In addition, none of the participants explicitly expressed a subscription to any particular pronunciation model or standard, but rather that they focused on intelligibility over native speaker accent accuracy. Through personal communications with the Swedish National Agency for Education (Skolverket) and a university professor of English at a teacher education programme, tendencies are observable of attempts at shifting responsibility for interpreting, understanding, and applying the syllabi, ultimately leaving individual teachers to uphold the demand of an equitable education through subjective interpretations of pronunciation teaching.  The implications of this study suggest that the field of pronunciation teaching in both Swedish and international context is still underdeveloped and in desperate need of further research. Whilst this study may have limited reach or impact on the field as such, it may serve as an indicator for the problem at large for teachers, researchers and educational agencies as well as promoting an awareness of issues in the equitability of pronunciation teaching. This paper may also serve as a basis for discussion in teacher teams or other educational opportunities for teachers on the development of coherent pronunciation constructs.