Satisfação do cliente na área da restauração

Dissertação de Mestrado , Ciências Económicas e Empresariais, Faculdade de Economia, Universidade do Algarve, 2008

Synthesis and evaluation of polymer nanoparticles for delivery in RPE cells

Ocular pathologies are among the most debilitating medical conditions affecting all segments of the population. Traditional treatment options are often ineffective, and gene therapy has the potential to become an alternative approach for the treatment of several pathologies. Methacrylate polymers have been described as highly biocompatible and are successfully used in medical applications. Due to their cationic nature, these polymers can be used to form polyplexes with DNA for its delivery. This work aims to study the potential of PDMAEMA (poly(2-(N,N’-dimethylamino)ethyl methacrylate)) as a non viral gene delivery system to the retina. The first part of this work aimed to study the potential for gene delivery of a previously synthesized PDMAEMA polymer of high molecular weight (354kDa). In the second part, we synthesized by RAFT a PDMAEMA with a lower molecular weight (103.3kDa) and similarly, evaluated its ability to act as a gene delivery vehicle. PDMAEMA/DNA polyplexes were prepared at 5, 7.5, 10, 12.5 and 20 nitrogen/phosphorous (N/P) ratio for the 354kDa PDMAEMA and at 5 and 7.5 for the 103.3kDa PDMAEMA. Dynamic light scattering and zeta potential measurements confirmed the nanosize and positive charge of polyplexes for all ratios and for both polymers. Both high and low Mw PDMAEMA were able to efficiently complex and protect DNA from DNase I degradation. Their cytotoxicity was evaluated using a non-retinal cell line (HEK293) and a retinal pigment epithelium (RPE) cell line (D407). We have found that cytotoxicity of the free polymer is concentration and time dependent, as expected, and negligible for all the concentrations of the PDMAEMA-DNA polyplexes. Furthermore, for the concentrations to be used in vivo, the 354kDa PDMAEMA showed no signs of inflammation upon injection in the intravitreal space of C57BL/6 mice. The transfection efficiency, as evaluated by fluorescence microscopy and flow cytometry, showed that the D407 retinal cells were transfected by polyplexes of both high and low Mw PDMAEMA, but with varied efficiency, which was dependent on the N/P ratio. Althogether, these results suggest that PDMAEMA is a feasible candidate for non-viral gene delivery to the retina, and this work constitutes the basis of further studies to elucidate the bottleneck in transfection and further optimization of the material.

An embodied approach to language: the role of bodily feedback in discourse processing

Theories of embodied cognition argue that language processing arises not from amodal symbols that redescribe sensorimotor and affective experiences, but from partial simulations (reenactments) of modality-specific states. Recent findings on processing of words and sentences support such a stance emphasizing that the role of the body in the domain of language comprehension should not be overlooked or dismissed. The present research was conducted to extend prior work in two important ways. First, the role of simulation was tested with connected discourse rather than words or sentences presented in isolation. Second, both “online” and “offline” measures of discourse comprehension were taken. In Experiments 1 and 2 participants’ facial postures were manipulated to show that preparing the body for processing of emotion-congruent information improves discourse comprehension. In Experiment 3 the direction of body posture was manipulated to show that implicit properties of simulations, such as spatial dimension or location, are at least somewhat involved in processing of large language segments such as discourse. Finally, in Experiments 4 and 5 participants’ body movement and body posture were manipulated to show that even understanding of language describing metaphorical actions physically impossible to perform involves constructing a sensorimotor simulation of the described event. The major result was that compatibility between embodiment and language strongly modulated performance effectiveness in experiments on simulation of emotion and metaphorical action. The effect of simulation on comprehension of discourse implying spatial dimension was fragile. These findings support an embodied simulation account of cognition suggesting that sensorimotor and affective states are at least partially implicated in “online” and “offline” discourse comprehension.

Tourism in Timor Leste: contributions to a sustainable development

The main goal of this thesis is to analyse tourism as a contributor to sustainable development, from a supply and demand perspective, in Timor-Leste, a destination not only “young” but also framed in an early stage of touristic development. From the supply side, studies 1 and 2 present a literature review on the concept of sustainable development, in small islands destinations, its weaknesses and vulnerabilities, and whose attractiveness lies in an integrated set of natural resources (attractive beaches, biodiversity, the coral triangle, a rich fauna and flora), its geographic location, tropical climate, socio cultural tradition and a Luso-Timorense cultural mix, that can meet the needs of important segments of the tourist demand. In these studies, the focus is on the residents’ attitudes in relation to sustainable development and, in particular, on the perceptions of local leaders about the importance of the operationalization of the concept. As Timor-Leste is a “young” destination, it is travel industry is still faced with several problems in order to be able to satisfy the demand and to meet the tourists’ motivations, preferences and requirements. In this perspective, the study 3 examines diaspora tourism, while a niche market, allowing for not only the entry of Timor-Leste in the international tourism market, but also an articulation between the people of Timor-Leste in the diasporas and the development of sustainable tourism . In general, the results show that the development of sustainable tourism cannot be successfully implemented without the involvement of all stakeholders such as residents, public sector (local government, policy makers, planners), private sector (tourism operators), and non-governmental organizations (NGOs) that include civil society, religious institutions, the academic community (the supply side), and Timorese diasporas tourists as consumers (the demand side).