The role of the mammalian DNA end-processing enzyme polynucleotide kinase 3'-phosphatase in spinocerebellar ataxia Type 3 pathogenesis

DNA strand-breaks (SBs) with non-ligatable ends are generated by ionizing radiation, oxidative stress, various chemotherapeutic agents, and also as base excision repair (BER) intermediates. Several neurological diseases have already been identified as being due to a deficiency in DNA end-processing activities. Two common dirty ends, 3'-P and 5'-OH, are processed by mammalian polynucleotide kinase 3'-phosphatase (PNKP), a bifunctional enzyme with 3'-phosphatase and 5'-kinase activities. We have made the unexpected observation that PNKP stably associates with Ataxin-3 (ATXN3), a polyglutamine repeat-containing protein mutated in spinocerebellar ataxia type 3 (SCA3), also known as Machado-Joseph Disease (MJD). This disease is one of the most common dominantly inherited ataxias worldwide; the defect in SCA3 is due to CAG repeat expansion (from the normal 14-41 to 55-82 repeats) in the ATXN3 coding region. However, how the expanded form gains its toxic function is still not clearly understood. Here we report that purified wild-type (WT) ATXN3 stimulates, and by contrast the mutant form specifically inhibits, PNKP's 3' phosphatase activity in vitro. ATXN3-deficient cells also show decreased PNKP activity. Furthermore, transgenic mice conditionally expressing the pathological form of human ATXN3 also showed decreased 3'-phosphatase activity of PNKP, mostly in the deep cerebellar nuclei, one of the most affected regions in MJD patients' brain. Finally, long amplicon quantitative PCR analysis of human MJD patients' brain samples showed a significant accumulation of DNA strand breaks. Our results thus indicate that the accumulation of DNA strand breaks due to functional deficiency of PNKP is etiologically linked to the pathogenesis of SCA3/MJD.

Annoyance and welfare costs from the presence of renewable energy power plants: an application of the contingent valuation method

Sustainability is frequently defined by its three pillars: economically viable, socially equitable, and environmentally bearable. Consequently the evaluation of the sustainability of any decision, public or private, requires information on these three dimensions. This paper focuses on social sustainability. In the context of renewable energy sources, the examination of social sustainability requires the analysis of not only the efficiency but also the equity of its welfare impacts. The present paper proposes and applies a methodology to generate the information necessary to do a proper welfare analysis of the social sustainability of renewable energy production facilities. This information is key both for an equity and an efficiency analysis. The analysis focuses on the case of investments in renewable energy electricity production facilities, where the impacts on local residents’ welfare are often significantly different than the welfare effects on the general population. We apply the contingent valuation method to selected facilities across the different renewable energy power plants located in Portugal and conclude that local residents acknowledge differently the damage sustained by the type, location and operation of the plants. The results from these case studies attest to the need of acknowledging and quantifying the negative impacts on local communities when assessing the economic viability, social equity and environmental impact of renewable energy projects.

Cost optimal strategies for the renovation of residential neighbourhoods towards energy and emissions neutrality – Rainha Dona Leonor case study

Buildings are one of the major consumers of energy in Europe. This makes them an important target when aiming to reduce the energy consumptions and carbon emissions. The majority of the European building stock has already some decades and so it needs renovation in order to keep its functionality. Taking advantage of these interventions, the energy performance of the buildings may also be improved. In Portugal the renovation techniques, both regarding energy efficiency measures as well as measures for the use of renewable energy sources, are normally planned at the building scale. It is important to explore the possibility of having large scale interventions, has it has been done in other countries, namely at neighbourhood scale with district energy system in order to optimize the results in terms of costs and environmental impact.

Promoting sustainability as a strategy to mitigate the effects of economic downturn on the construction industry

During the recent years followed by the Global Financial Crisis (GFC), most of business and industries around the globe have been hardly hit to the limit that it still struggling to survive, suffering from the crisis financial consequences. For instance, in the construction industry; many construction projects have been suspended or totally cancelled. Nevertheless, among this dilemma, a call has been raised to use the sustainable practices to mitigate the effects of the GFC on construction industry. For the first look, it seems that there is contradiction since the sustainable solutions are often associated with an increase in the initial cost, undoubtedly, the sustainable practices have many advantages in both economic and environment aspects, however, the question which needs to be addressed here is, to what extent using such sustainable practices can mitigate the negative effects of the economic downturn on construction industry. Therefore, it is a challenging argument for using such sustainable construction from its economic perspective, however, this paper is aiming to present the economical benefits of sustainable practices in construction industry, and trying to clear the doubt of the high initial costs of the sustainable construction through studying the life cycle benefit of green building.

The Warburg effect in mycobacterial granulomas is dependent on the recruitment and activation of macrophages by interferon-γ

Granulomas are the hallmark of mycobacterial disease. Here, we demonstrate that both the cell recruitment and the increased glucose consumption in granulomatous infiltrates during Mycobacterium avium infection are highly dependent on interferon-y (IFN-y). Mycobacterium avium-infected mice lacking IFN-y signalling failed to developed significant inflammatory infiltrations and lacked the characteristic uptake of the glucose analogue fluorine-18-fluorodeoxyglucose (FDG). To assess the role of macrophages in glucose uptake we infected mice with a selective impairment of IFN-y signalling in the macrophage lineage (MIIG mice). Although only a partial reduction of the granulomatous areas was observed in infected MIIG mice, the insensitivity of macrophages to IFN-y reduced the accumulation of FDG. In vivo, ex vivo and in vitro assays showed that macrophage activated by IFN-y displayed increased rates of glucose uptake and in vitro studies showed also that they had increased lactate production and increased expression of key glycolytic enzymes. Overall, our results show that the activation of macrophages by IFN-y is responsible for the Warburg effect observed in organs infected with M. avium.

Study of the tightness of the metallization used on top of a thin film capacitor cell

Dissertação de mestrado integrado em Engenharia de Materiais

The influence of the Palestinian sociocultural values in shaping the vernacular architecture of Nablus city

Recently, environmental architecture and sustainable construction has been ranked on top of the worldâ s interests. Making use of natural resources helps in reducing energy consumption and costs associated with the operation of buildings. The current architectural approaches and designs in Palestine are far away from environmental concepts, copying and simulating abroad approaches, without taking into account the culture, climate, and inhabitant's needs. On the contrast, vernacular architecture has achieved environmental concepts and has given suitable approaches and samples - without any need to simulate or copy - which come from people and land. This paper discusses how the Palestinian socio-cultural context shaped the residential vernacular architecture in Palestine, taking the old city of Nablus as a case-study. The research concept depends on analysing and trying to understand the effect of the socio-cultural context on vernacular architecture and trying to reach some rules or understandings of how it works in order to reach a modern environmental dwelling that is suitable to this concept. The research method goes through analysing study cases from the traditional architecture models and the Nablus city is selected as a case study. This analytical and qualitative method can lead to deep understanding for how to benefit from vernacular architecture in Palestine in finding the future environmental residential construction. One of the main findings of this research is to set general and special rules for building sustainable buildings in Palestine from the socio-cultural point view, in order to be a reference for designers, stakeholders, ministry of planning, and municipalities.

Toxicity induced by a metal mixture (Cd, Pb and Zn) in the yeast Pichia kudriavzevii: The role of oxidative stress

The present work aims to contribute for the elucidation of the role of oxidative stress in the toxicity associated with the exposure of Pichia kudriavzevii to multi-metals (Cd, Pb and Zn). Cells of the non-conventional yeast P. kudriavzevii exposed for 6 h to the action of multi-metals accumulated intracellular reactive oxygen species (ROS), evaluated through the oxidation of the probe 2,7-dichlorodihydrofluorescein diacetate. A progressive loss of membrane integrity (monitored using propidium iodide) was observed in multi-metal-treated cells. The triggering of intracellular ROS accumulation preceded the loss of membrane integrity. These results suggest that the disruption of membrane integrity can be attributed to the oxidative stress. The exposure of yeast cells to single metal showed that, under the concentrations tested, Pb was the metal responsible for the induction of the oxidative stress. Yeast cells coexposed to an antioxidant (ascorbic acid) and multi-metals did not accumulate intracellular ROS, but loss proliferation capacity. Together, the data obtained indicated that intracellular ROS accumulation contributed to metal toxicity, namely for the disruption of membrane integrity of the yeast P. kudriavzevii. It was proposed that Pb toxicity (the metal responsible for the toxic symptoms under the conditions tested) result from the combination of an ionic mechanism and the intracellular ROS accumulation.

Determination of the importance of iron acquisition for Staphylococcus epidermidis biofilm survival in human blood

Dissertação de mestrado em Bioengenharia