The objectives were to analyze one-year survival and mortality predictors in patients with fracture of the proximal femur (low/moderate trauma). A prospective cohort was formed by inviting all patients hospitalized in the Orthopedic Ward of the second largest hospital in Portugal (May 2008-April 2009). Survival was assessed at 3, 6, 9, and 12 months after fracture and related to demographic factors, lifestyle, and clinical history, as well as to data from medical records (fracture type, surgery date, surgical treatment, and preoperative risk). Of the 340 patients hospitalized, 252 were included (78.9% women). Mortality at 3, 6, 9, and 12 months was 21.2%, 25%, 28.8%, and 34.6% for men and 7.8%, 13.5%, 19.2%, and 21.4% for women, respectively. Predictors of death were male gender (HR = 2.54; 95%CI: 1.40-4.58), ASA score III/IV vs. I/II (HR = 1.95; 95%CI: 1.10-3.47), age (HR = 1.06; 95%CI: 1.03-1.10), and delay in days to surgery (HR = 1.07; 95%CI: 1.03-1.12). Factors related to death were mainly related to patients' characteristics at admission.

In this work, we present a procedure to grow single phase SnS thin films consisting on the annealing of RF magnetron sputtered SnS2 precursors . A series of samples was produced by rapid thermal processing of precursors deposited both on bare and Mo coated glass. For those samples the time at maximum temperature and heating rate were varied

In this work we have studied the influence of the Ar working pressure, substrate temperature, low power plasma irradiation and partial pressure of hydrogen in the RF-magnetron sputtering of indium tin oxide (ITO) thin films on glass substrates. This work aims at identifying the best conditions to achieve good quality ITO film at low temperature. Four sets of samples were prepared which were characterized by scanning electron microscopy, X-ray diffraction (XRD), Van der Pauw, transmittance and absorbance measurements

In the present work, we present a process to grow tin diselenide thin films by selenization at a maximum temperature of 470 ºC, of tin metallic precursor layers deposited by dc magnetron sputtering. For this maximum temperature, disklike grain morphologies were observed. Prominent XRD reflections at 2θ= 30.75º, 40.10º and 47.72º and vibration modes located at 119 cm-1 and 185 cm-1 were observed. These results allowed concluding that the dominant phase is SnSe2. The composition analysis, done by energy dispersive spectroscopy (EDS), showed that the films were close to being stoichiometric SnSe2 with a Se to Sn ratio of 1.95. Photoluminescence characterization was performed and revealed a dominant band at 0.874 eV and two other bands at ~0.74 and 1.08 eV with a lower relative intensities. The observed radiative transitions depend critically on the temperature.

The prevalence of poisoning events due to harmful algal blooms (HABs) has declined during the last two decades through monitoring programs and legislation, implemented mainly for bivalves. However, new toxin vectors and emergent toxins pose a challenge to public health. Several locations on the Portuguese coast were surveyed between 2009 and 2010 for three distinct biotoxin groups [saxitoxin (PST), spirolide (SPX) and okadaic acid (OA)], in 14 benthic species of mollusks and echinoderms. Our main goals were to detect new vectors and unravel the seasonal and geographical patterns of these toxins. PSTs were analyzed by the Lawrence method, SPXs by LC-MS/MS, and OA by LC-MS/MS and UPLC-MS/MS. We report 16 new vectors for these toxins in the North Atlantic. There were differences in toxin contents among species, but no significant geographical or seasonal patterns were found. Our results suggest that legislation should be adjusted to extend the monitoring of marine toxins to a wider range of species besides edible bivalves.

Toxic cyanobacterial blooms are documented worldwide as an emerging environmental concern. Recent studies support the hypothesis that microcystin-LR (MC-LR) and cylindrospermopsin (CYN) produce toxic effects in crop plants. Lettuce (Lactuca sativa L.) is an important commercial leafy vegetable that supplies essential elements for human nutrition; thus, the study of its sensitivity to MC-LR, CYN and a MC-LR/CYN mixture is of major relevance. This study aimed to assess the effects of environmentally relevant concentrations (1, 10 and 100 µg/L) of MC-LR, CYN and a MC-LR/CYN mixture on growth, antioxidant defense system and mineral content in lettuce plants. In almost all treatments, an increase in root fresh weight was obtained; however, the fresh weight of leaves was significantly decreased in plants exposed to 100 µg/L concentrations of each toxin and the toxin mixture. Overall, GST activity was significantly increased in roots, contrary to GPx activity, which decreased in roots and leaves. The mineral content in lettuce leaves changed due to its exposure to cyanotoxins; in general, the mineral content decreased with MC-LR and increased with CYN, and apparently these effects are time and concentration-dependent. The effects of the MC-LR/CYN mixture were almost always similar to the single cyanotoxins, although MC-LR seems to be more toxic than CYN. Our results suggest that lettuce plants in non-early stages of development are able to cope with lower concentrations of MC-LR, CYN and the MC-LR/CYN mixture; however, higher concentrations (100 µg/L) can affect both lettuce yield and nutritional quality.

The formulation of magnetic ionic liquids (MILs) or organic salts based on lanthanides as anions has been explored. In this work, a set of choline-family-based salts, and two other, different cation families, were combined with Gadolinium(III) and Terbium(III) anions. Synthetic methodologies were previously optimized, and all organic salts were obtained as solids with melting temperatures higher than 100 °C. The magnetic moments obtained for the Gd(III) salts were, as expected, smaller than those obtained for the Tb(III)-based compounds. The values for Gd(III) and Tb(III) magnetic salts are in the range of 6.55–7.30 MB and 8.22–9.34 MB, respectively. It is important to note a correlation between the magnetic moments obtained for lanthanides, and the structural features of the cation. The cytotoxicity of lanthanide-based salts was also evaluated using 3T3, 293T, Caco2, and HepG2 cells, and it was revealed that most of the prepared compounds are not toxic.

The potential of WW domains as proteins scaffolds for the development of purification affinity reagents has been explored. Minimal versions of native human Pin1 (hPin1_WW) and human YAP65 (hYAP65_WW) WW domains were produced through chemical synthesis using solid-phase peptide synthesis. After purification, characterization by mass spectrometry and circular dichroism demonstrated, respectively, the correct molecular mass, and the correct folding of WW domains with thermal stability comparable to the extended version expressed biologically. These peptides were further immobilized in chromatographic supports. Adsorbents bearing hPin1_WW captured phosphorylated peptides and proteins, whereas those modified with hYAP65_WW captured Proline-rich peptides. In both cases the binding and elution of the target peptides was achieved under mild conditions. Encouraged by these results, a näive library based on the sequence of WW prototype was designed and generated. The library was further evolved in vitro through phage and ribosome display methodologies against Human Serum Albumin (HSA) and Immunoglobulin G (IgG). The phage display library leads to the identification of CW3S (a WW domain derived from Clone 3) as a potential HSA binder. This ligand was produced biologically in co-expression with GFP, and was also chemically synthesized. The affinity between CW3S and HSA was determined as Ka of 8.37x106 M-1 (KD=119nM) by ELISA. The chemically synthesized peptide was characterized by circular dichroism showing the folding and thermal stability similar to other native WW domains. This peptide was also immobilized in agarose and captured HSA (0.291μg protein/mg support at 4°C). This work strongly demonstrated the robustness of the WW domains to withstand modifications and mutations, therefore possessing a clear potential for application as a protein scaffold.

BACKGROUND: Building reference libraries of DNA barcodes is relatively straightforward when specifically designed primers are available to amplify the COI-5P region from a relatively narrow taxonomic group (e.g. single class or single order). DNA barcoding marine communities have been comparatively harder to accomplish due to the broad taxonomic diversity and lack of consistently efficient primers. Although some of the so-called "universal" primers have been relatively successful, they still fail to amplify COI-5P of many marine animal groups, while displaying random success even among species within each group. Here we propose a new pair of primers designed to enhance amplification of the COI-5P region in a wide range of marine organisms. RESULTS: Amplification tests conducted on a wide range of marine animal taxa, rendered possible the first--time sequencing of DNA barcodes from eight separated phyla (Annelida, Arthropoda, Chordata, Cnidaria, Echinodermata, Mollusca, Nemertea and Platyhelminthes), comprising a total of 14 classes, 28 orders, 57 families, 68 genus and 76 species. CONCLUSIONS: These primers demonstrated to be highly cost-effective, which is of key importance for DNA barcoding procedures, such as for building comprehensive DNA barcode libraries of marine communities, where the processing of a large numbers of specimens from a wide variety of marine taxa is compulsory.

A diabetes tipo 2 é uma das doenças crónicas mais comuns no mundo, cuja prevalência continua a aumentar. Em 2045 estima-se que esta doença afete cerca de 629 milhões de pessoas no mundo. A diabetes tipo 2 é caracterizada pela resistência periférica à insulina, o anormal metabolismo hepático de glucose e a falha progressiva das células beta do pâncreas. Existem vários fármacos disponíveis para o tratamento da diabetes tipo 2 contudo, com o tempo, o controlo da glucose deteriora-se progressivamente e, mesmo com ajustes na medicação, uma proporção considerável de indivíduos continua pouco controlado. Deste modo, é essencial o desenvolvimento de novas estratégicas terapêuticas para o controlo da diabetes tipo 2. Nos últimos anos, o corpo carotídeo (CB), um quimoreceptor periférico que deteta alterações de O2, CO2 e pH no sangue, tem também sido descrito como um sensor metabólico envolvido no controlo da homeostasia energética. De facto, foi descrito que o CB está envolvido na génesis da resistência à insulina e hipertensão induzidos pelas dietas hipercalóricas. Este trabalho teve como objetivo avaliar o papel do CB no controlo da homeostasia da glucose e investigar uma possível metodologia para modular a atividade do CB com o intuito de encontrar um novo tratamento para a diabetes tipo 2. O capítulo I introduz conceitos gerais na diabetes tipo 2, tais como, a sinalização da insulina, a homeostasia da glucose e as opções terapêuticas para o tratamento da diabetes tipo 2. Para além disso, conceitos relacionados com a função do CB e o papel do ATP e da adenosina na neurotransmissão do CB, bem como, o seu papel como sensor metabólico serão também abordados. No capítulo II estão descritos os objetivos gerais e específicos do presente trabalho. No capítulo III demonstrou-se que a ressecção do nervo do seio carotídeo (CSN), o nervo sensitivo do CB, restaura a sensibilidade à insulina em dois modelos animais de prediabetes, um efeito que se mantém com a continuação da administração das dietas hipercalóricas. Para além disso, foi observado que a ressecção do CSN normaliza a atividade do sistema nervo simpático, a pressão arterial, a função endotelial, o perfil lipídico e os níveis plasmáticos de glucose e insulina. Foi também descrito que o mecanismo pelo qual é restaurada a homeostasia da glucose envolve uma melhoria na captação de glucose no fígado e no tecido adiposo perientérico bem como, o restauro das vias de sinalização da insulina no músculo esquelético e no tecido adiposo. No capítulo IV foi descrito que a modulação bioelectrónica do CSN através da aplicação de uma corrente alternada de alta frequência (KHFAC) restaura a sensibilidade à insulina e a tolerância à glucose num modelo animal de diabetes tipo 2. Para além disso, observou-se que estes efeitos são reversíveis após o término do estímulo elétrico de alta frequência. Assim, este trabalho suporta o potencial terapêutico da medicina bioelectrónica na diabetes tipo 2. Uma abordagem farmacológica poderá também ser utilizada para modular a atividade do CSN. Desta forma, no capítulo V foi estudado o papel do ATP e da adenosina na atividade do CSN no basal e em resposta à hipóxia. Observou-se que a frequência de descarga do CSN está aumentada num modelo animal de prediabetes, sendo este efeito modulado pelo ATP e pela adenosina. Tendo em conta que a adenosina contribui mais do que o ATP para gerar atividade do CSN na hipóxia moderada, enquanto que o ATP tem um papel preponderante durante a hipóxia intensa e, sabendo que hipóxias intensas são menos frequentes em situações fisiológicas, é sugerido que a modulação da sinalização do ATP no CB poderá ser um alvo terapêutico para o tratamento da diabetes tipo 2. Finalmente, no capítulo VI, é apresentada uma discussão geral e integrada da presente tese de Doutoramento. Em conclusão, os resultados apresentados neste trabalho contribuem para reforçar que a modulação do CB/CSN representa uma nova estratégia terapêutica na diabetes tipo 2.