Design,optimization and in vivo evaluation of non-viral vectors for gene therapy. Applications in ocular disorders

227 págs.

Analysis of SUMOylated proteins using SUMO-traps

6 p. [+ 7 p. Supplementary Information]

Keep an "eye" on the text: The Use of Self-mention in the Academic Writing of Experts

One of the most controversial inquiries in academic writing is whether it is admissible to use first person pronouns in a scientific paper or not. Many professors discourage their students from using them, rather favoring a more passive tone, and thus causing novices to avoid inserting themselves into their texts in an expert-like manner. Abundant research, however, has recently attested that negotiation of identity is plausible in academic prose, and there is no need for a paper to be void of an authorial identity. Because in the course of the English Studies Degree we have received opposing prompts in the use of I, the aim of this dissertation is to throw some light upon this vexed issue. To this end, I compiled a corpus of 16 Research Articles (RAs) that comprises two sub-corpora, one featuring Linguistics RAs and the other one Literature RAs, and each, in turn, consists of articles written by American and British authors. I then searched for real occurrences of I, me, my, mine, we, us, our and ours, and studied their frequency, rhetorical functions and distribution along each paper. The results obtained certainly show that academic writing is no longer the faceless prose that it used to be, for I is highly used in both disciplines and varieties of English. Concerning functions, the most typically used roles were the use of I to take credit for the writer’s research process, and also those involving plural forms. With respect to the spatial disposition, all sections welcomed first person pronouns, but the Method and the Results/Discussion sections seem to stimulate their appearance. On the basis of these findings, I suggest that an L2 writing pedagogy that is mindful not only of the language proficiency, but also of the students’ own identity may have a beneficial effect on the composition of their texts.

Structural studies of the hipersaline adaptation of proteins belonging to halophilic archaea

183 p.

Proteomic Analysis of the Ubiquitin Landscape in the Drosophila Embryonic Nervous System and the Adult Photoreceptor Cells

Background Ubiquitination is known to regulate physiological neuronal functions as well as to be involved in a number of neuronal diseases. Several ubiquitin proteomic approaches have been developed during the last decade but, as they have been mostly applied to non-neuronal cell culture, very little is yet known about neuronal ubiquitination pathways in vivo. Methodology/Principal Findings Using an in vivo biotinylation strategy we have isolated and identified the ubiquitinated proteome in neurons both for the developing embryonic brain and for the adult eye of Drosophila melanogaster. Bioinformatic comparison of both datasets indicates a significant difference on the ubiquitin substrates, which logically correlates with the processes that are most active at each of the developmental stages. Detection within the isolated material of two ubiquitin E3 ligases, Parkin and Ube3a, indicates their ubiquitinating activity on the studied tissues. Further identification of the proteins that do accumulate upon interference with the proteasomal degradative pathway provides an indication of the proteins that are targeted for clearance in neurons. Last, we report the proof-of-principle validation of two lysine residues required for nSyb ubiquitination. Conclusions/Significance These data cast light on the differential and common ubiquitination pathways between the embryonic and adult neurons, and hence will contribute to the understanding of the mechanisms by which neuronal function is regulated. The in vivo biotinylation methodology described here complements other approaches for ubiquitome study and offers unique advantages, and is poised to provide further insight into disease mechanisms related to the ubiquitin proteasome system.