The Evolution of Function in the Rab family of Small GTPases

Dissertation presented to obtain the PhD degree in Computational Biology.

Immunohistochemistry detection of putative miR-200c and miR-203 targets in breast cancer patients

Part of this thesis will be published in the following: Gomes, B.C., Santos, B. 2015. Methods for studying microRNAs expression and their targets in formalin-fixed, paraffin-embedded (FFPE) breast cancer tissues. In Methods in Molecular Biology: Cancer Drug Resistance (Rueff, J. & Rodrigues, A.S. eds), Springer Protocols.

Establishing guidelines for obtaining optimal cell sources to use in tendon tissue engineering strategies

Cell-based approaches in tissue engineering (TE) have been barely explored for the treatment of tendon and ligament (T/L) tissues, requiring the establishment of a widely available cell source with tenogenic potential. As T/L cells are scarce, stem cells may provide a good alternative. Understanding how resident cells behave in vitro, might be useful for recapitulating the tenogenic potential of stem cells for tendon TE applications. Therefore, we propose to isolate and characterize human T/L-derived cells (hTDCs and hLDCs) and compare their regenerative potential with stem cells from adipose tissue (hASCs) and amniotic fluid (hAFSCs)(1). T/L cells were isolated using different procedures and stem cells isolated as described elsewhere(1). Moreover, T/L cells were stimu- lated into the three mesenchymal lineages, using standard differentia- tion media. Cells were characterized for the typical stem cell markers as well as T/L related markers, namely tenascin-C, collagen I and III, decorin and scleraxis, using different complementary techniques such as real time RT-PCR, immunocytochemistry and flow cytometry. No differences were observed between T/L in gene expression and protein deposition. T/L cells were mostly positive for stem ness markers (CD73/CD90/CD105), and have the potential to differentiate towards osteogenesis, chondrogenesis and adipogenesis, demonstrated by the positive staining for AlizarinRed, SafraninO, ToluidineBlue and OilRed. hASCs and hAFSCs exhibit positive expression of all tenogenic mark- ers, although at lower levels than hTDCs and hLDCs. Nevertheless, stem cells availability is key factor in TE strategies, despite that it’s still required optimization to direct their tenogenic phenotype.

Electrically tunable resonant scattering in fluorinated bilayer graphene

Adatom-decorated graphene offers a promising new path towards spintronics in the ultrathin limit. We combine experiment and theory to investigate the electronic properties of dilutely fluorinated bilayer graphene, where the fluorine adatoms covalently bond to the top graphene layer. We show that fluorine adatoms give rise to resonant impurity states near the charge neutrality point of the bilayer, leading to strong scattering of charge carriers and hopping conduction inside a field-induced band gap. Remarkably, the application of an electric field across the layers is shown to tune the resonant scattering amplitude from fluorine adatoms by nearly twofold. The experimental observations are well explained by a theoretical analysis combining Boltzmann transport equations and fully quantum-mechanical methods. This paradigm can be generalized to many bilayer graphene-adatom materials, and we envision that the realization of electrically tunable resonance may be a key advantage in graphene-based spintronic devices.

Numerical modeling and assessment of the thermal environment in an industrial premise of automotive components

Dissertação de mestrado em Engenharia Industrial

The influence of kaolin application on key metabolic pathways associated with grape berry quality: a molecular and biochemical analysis

Dissertação de mestrado em Biologia Molecular, Biotecnologia e Bioempreendedorismo em Plantas

Lipids under stress - a lipidomic approach for the study of mood disorders

The emerging field of lipidomics has identified lipids as key players in disease physiology. Their physicochemical diversity allows precise control of cell structure and signaling events through modulation of membrane prop- erties and trafficking of proteins. As such, lipids are important regulators of brain function and have been implicated in neurodegenerative and mood disorders. Importantly, environmental chronic stress has been associated with anxiety and depression and its exposure in rodents has been extensively used as a model to study these diseases. With the accessibility to modern mass- spectrometry lipidomic platforms, it is now possible to snapshot the extensively interconnected lipid network. Here, we review the fundamentals of lipid biology and outline a framework for the interpretation of lipidomic studies as a new approach to study brain pathophysiology. Thus, lipid profiling provides an exciting avenue for the identification of disease signatures with important implications for diagnosis and treatment of mood disorders.

Safety management of cranes on construction sites in Portugal: Critical factors of safety

The evolution of the construction caused a need to use more effective equipments, capable of meeting the increasingly demanding deadlines for the completion of works. In this context, the safety and efficiency of equipment have become key aspects in order to optimize the execution time of the works, as well as reducing labor costs and loss of materials. With the evolution of construction and construction processes, cranes have come to represent a signal of the construction of buildings, revealing to be, in most of the cases, the main equipment of construction sites. Currently, some engineers revels some apprehension regarding the use and handling of cranes which is natural and acceptable, since an equipment failure can lead to serious or fatal accidents. The factors affecting safety management of the cranes in construction sites were investigated, identified, classified and evaluated according to their degree of importance, through interviews with representatives of the general contractors of a set of selected construction sites.

Human Tissue Kallikrein Activity in Angiographically Documented Chronic Stable Coronary Artery Disease

AbstractBackground:Human tissue kallikrein (hK1) is a key enzyme in the kallikrein–kinin system (KKS). hK1-specific amidase activity is reduced in urine samples from hypertensive and heart failure (HF) patients. The pathophysiologic role of hK1 in coronary artery disease (CAD) remains unclear.Objective:To evaluate hK1-specific amidase activity in the urine of CAD patientsMethods:Sixty-five individuals (18–75 years) who underwent cardiac catheterism (CATH) were included. Random midstream urine samples were collected immediately before CATH. Patients were classified in two groups according to the presence of coronary lesions: CAD (43 patients) and non-CAD (22 patients). hK1 amidase activity was estimated using the chromogenic substrate D-Val-Leu-Arg-Nan. Creatinine was determined using Jaffé’s method. Urinary hK1-specific amidase activity was expressed as µM/(min · mg creatinine) to correct for differences in urine flow rates.Results:Urinary hK1-specific amidase activity levels were similar between CAD [0.146 µM/(min ·mg creatinine)] and non-CAD [0.189 µM/(min . mg creatinine)] patients (p = 0.803) and remained similar to values previously reported for hypertensive patients [0.210 µM/(min . mg creatinine)] and HF patients [0.104 µM/(min . mg creatinine)]. CAD severity and hypertension were not observed to significantly affect urinary hK1-specific amidase activity.Conclusion:CAD patients had low levels of urinary hK1-specific amidase activity, suggesting that renal KKS activity may be reduced in patients with this disease.

Ostracodes (Crustacea) from Cananéia-Iguape estuarine/lagoon system and geographical distribution of the mixohaline assemblages in southern and southeastern Brazil

The ostracode assemblages from Cananéia-Iguape estuarine/lagoon system (southernmost State of São Paulo) are here discussed in detail for the first time. Thirty-four sites, approximately 1 km equidistant, were sampled along the system, including the Cananéia Sea, Pequeno Sea, Cubatão Sea, Ribeira de Iguape River and Itapitangui River. The ostracodes throughout this area have poor assemblages, with a total of 662 specimens of dead and living organisms. The majority of the ostracode fauna is composed of euryhaline species, as follows: Cyprideis multidentata Hartmann, 1955 (174 specimens), Minicythere heinii Ornellas, 1974 (54 specimens), Tanella gracilis Kingma, 1948 (96 specimens) and Whatleyella sanguinettiae Coimbra, Carreño & Ferron, 1994 (226 specimens). Although there are few studies on the Brazilian mixohaline ostracode faunas, including the euryhaline marginal marine taxa, the published data show that the group is best known in the south and southeast regions. Based on this review and with the new data presented in this paper, the geographical distribution of eight mixohaline key species in southern and southeastern Brazil is also discussed.

Bird community in an Araucaria forest fragment in relation to changes in the surrounding landscape in Southern Brazil

The dynamics of the bird community in a small forest fragment was evaluated along seven years in relation to changes in the surrounding landscape. The study area is an Araucaria forest fragment in Southern Brazil (state of Paraná). The sampling period covered the years 1988 through 1994 and the mark-release-recapture method was utilized. The landscape analysis was based on Landsat TM images, and changes in exotic tree plantations, native forest, open areas (agriculture, pasture, bare soil, and abandoned field), and "capoeira"(native vegetation < 2 m) were quantified. The relationship between landscape changes and changes in abundance diversity of forest birds, open-area birds, forest-edge birds, and bamboo specialists was evaluated. Richness estimates were run for each year studied. The richness recorded in the study area comprised 96 species. The richness estimates were 114, 118 and 110 species for Chao 1, Jackknife 1 and Bootstrap, respectively. The bird community varied in species richness, abundance and diversity from year to year. As for species diversity, 1991, 1993 and 1994 were significantly different from the other years. Changes in the landscape contributed to the increase in abundance and richness for the groups of forest, open-area and bamboo-specialist species. An important factor discussed was the effect of the flowering of "taquara" (Poaceae), which contributed significantly to increasing richness of bamboo seed eaters, mainly in 1992 and 1993. In general, the results showed that landscape changes affected the dynamics and structure of the bird community of this forest fragment over time, and proved to have an important role in conservation of the avian community in areas of intensive forestry and agricultural activities.

Why did London become the main money market? Monetary Policy, Arbitrage and European Money Market Integration in 18th century

How did the leading capital market start to attract international bullion? Why did London become the main money market? Monetary regulations, including the charges for minting money and the restrictions on bullion exchange, have played the key role in defining the direction of the flow of international bullion. Countries that abolished minting charges and permitted the free movement of bullion were able to attract international bullion, and countries that applied minting taxes suffered an outflow of bullion. In these cases monetary authorities tried to limit bullion movement through prohibitions on domestic bullion exchange at a free price, and tariffs and quantitative restrictions on bullion exports. The paper illustrates the logic of international monetary flow in the 18th century, using empirical evidence for England, France and Spain. The first section defines and measures monetary policy, and the second section introduces minting charges into the arbitrage equation in order to explain the logic of bullion flow between the pairs of nations England-France, England-Spain and France-Spain. The conclusion emphasises the importance of monetary policy in the creation of leading money markets.

Polysemy and Olympic audiences: lessons for sport marketing

Paper presented at the 2000 seminar of the International Chair in Olympism. Key topics of the seminar included references to the multiple narratives, embedded genres and layered symbols that are possible in an Olympic context.

Polisèmia i públic olímpic : lliçons de màrqueting esportiu

Ponència presentada al seminari de la Càtedra Internacional d'Olimpisme l'any 2000. Els temes principals del seminari inclouen les referències a la multiplicitat de discursos, la fixació de gèneres i la superposició de símbols que són possibles en el context olímpic.

Serum anticholinergic activity and postoperative cognitive dysfunction in elderly patients

Background: Cerebral cholinergic transmission plays a key role in cognitive function and anticholinergic drugs are associated with impaired cognitive functions [1]. In the perioperative phase many substances with anticholinergic effects are administered and disturbed cholinergic transmission is a hypothetical cause of postoperative cognitive dysfunction (POCD). Serum anticholinergic activity (SAA; pmol/ml) may be measured as a summary marker of anticholinergic activity in an individual patient's blood. We hypothesised that an increase in SAA from preoperatively to one week postoperatively is associated with POCD in elderly patients. Methods: Thirty-two patients aged >65 yrs undergoing elective major surgery under standardized general anaesthesia (thiopental, sevoflurane, fentanyl) were investigated. Cognitive functions were measured preoperatively and 7 days postoperatively using the extended version of the Consortium to Establish a Registry for Alzheimer's Disease - Neuropsychological Assessment Battery. POCD was defined as a postoperative decline >1 z-score in at least 2 cognitive domains. SAA was measured preoperatively and 7 days postoperatively at the time of cognitive testing. Results: 50% of the investigated patients developed POCD. There were no statistically significant differences between patients with and without POCD regarding age, education, baseline cognitive function, duration of anaesthesia, SAA preoperatively (median (range) 1.0 (0.3 to 5.0) vs 1.5 (0.4 to 5.0), SAA 7 days postoperatively (median (range) 1.3 (0.1 to 7.0) vs 1.4 (0.6 to 5.5) or changes in SAA (median (range) 0.1 (-1.6 to 2.2) vs 0.2 (-1.4 to 2.8). The variability of SAA in individual patients was considerable and marked changes in SAA between the two examinations were observed in some patients. However, there was no significant relationship between changes in SAA and changes in cognitive function. Conclusion: In this preliminary analysis of a small group of patients, changes in SAA in the perioperative phase were highly variable. SAA was not associated with POCD suggesting that POCD is not simply caused by anticholinergic medications administered in the perioperative phase. A further analysis of a larger group of patients is in progress.