Determinação e aplicações dos frutooligossacarídeos de Yacon (Smallanthus sonchifolius)

Torna-se cada vez mais importante fortalecer o vínculo entre o setor agrário, as universidades e o setor industrial. Através de estudos científicos pode-se aprimorar a produção agrícola, valorizar comercialmente os produtos manufaturados e permitir usos biotecnológicos anteriormente indisponíveis. Este trabalho ambicionou gerar dados que incentivassem e permitissem o suporte técnico-científico adequado ao pequeno e médio agricultor do litoral da Grande-Florianópolis, no estado de Santa Catarina (Brasil) viabilizando a exploração comercial das raízes de yacon (Smallanthus sonchifolius). Primeiro, o enfoque agronómico que objetivou ofertar dados científicos da adequação climática do plantio de yacon na região do litoral de Florianópolis (SC) com a finalidade de otimizar a produção e rentabilizá-la. Por cromatografia líquida de alta eficiência foram quantificados os valores de frutooligossacarídeos (fração de interesse, alimento funcional com efeito prebiótico), frutose, glucose e sacarose e avaliaram-se os efeitos diretos dos fatores climáticos na produção fazendo uso da análise multivariada para sua interpretação. Os resultados mostram que a produção de FOS não consegue atingir valores similares aos de outras regiões onde o yacon é explorado economicamente. Igualmente a produção de FOS parece ter uma grande dependência do clima, especialmente da amplitude térmica e da chuva. O segundo enfoque foi a possibilidade de exploração comercial, através de secagem em estufa. O processo de secagem, especialmente em amostras produzidas em condições climatológicas mais drásticas induzem modificações provocadas pela reacção de Maillard, como foi verificado por estudos de GCxGC-Tof-Ms e ESI-MS. O terceiro enfoque foi biotecnológico e consistiu em aproveitar frutooligossacarídeos comerciais para utilizá-los como matriz polimérica para produção de estruturas em escala nano e micrométrica através da eletropulverização. Foram observadas, nesta etapa, os aspetos morfológicos e as características das partículas formadas e a sua capacidade de servir como veículo transportador de compostos com características anti-oxidantes.

A proteomic study of the effects of nutrient restriction in utero on postnatal metabolism

It is widely recognized that protein restriction in utero may cause metabolic and endocrine adaptations, which may be of benefit to the neonate on a short-term basis but may cause adverse long-term conditions such as obesity, Type 2 diabetes, metabolic syndrome, hypertension and cardiovascular diseases. Adequate foetal and early post natal nutrient and energy supply is therefore essential for adult animal health, performance and life span. In this project it was investigated the progressive adaptations of the hepatic proteome in male mink offspring exposed to either a low protein (FL) or an adequate protein (FA) diet in utero fed either on a low protein (LP) or on an adequate (AP) diet from weaning until sexual maturity. Specifically, the aim was to determine the metabolic adaptations at selected phases of the animal’s first annual cycle and establish the metabolic priorities occurring during those phases. The three different morphological stages studied during the first year of development included, end of bone growth at 4 months of age, maximal fat accretion at 6 months of age and sexual maturity at 12 months of age. A reference proteome of mink liver coming from these different animal groups were generated using 2D electrophoresis coupled to MALDI-TOF analysis and the way in which dietary treatment affect their proteome was established. Approximately 330 proteins were detected in the mink liver proteome. A total of 27 comparisons were carried out between all different animal groups which resulted in 20 differentially expressed proteins. An extensive survey was conducted towards the characterization of these proteins including their subcellular localization, the biological processes in which they are involved and their molecular functions. This characterization allowed the identification of proteins in various processes including the glycolysis and fatty acid metabolism. The detailed analysis of the different dietary treatment animal groups was indicative of differences in metabolism and also to changes associated with development in mink.

Compilation of Periodic Histories from 1 April 1979 to 31 December 1989

This document covers operations, training, organization, key personnel, recognition and awards between 1979 and 1989.

Translating proteomics into environmental health : biomarkers discovery for tobacco smoke-induced biological damage

Tese de doutoramento, Biologia (Biologia Molecular), Universidade de Lisboa, Faculdade de Ciências, 2015

Influence of environmental variables on the predatory sucess of the iberian wolf in the north of Portugal

Tese de mestrado em Biologia da Conservação, apresentada à Universidade de Lisboa, através da Faculdade de Ciências, 2016

Characterization of metabolites of sweroside in rat urine using ultra-high-performance liquid chromatography combined with electrospray ionization quadrupole time-of-flight tandem mass spectrometry and NMR spectroscopy

Sweroside, a major active iridoid in Swertia pseudochinensis Hara, is recognized as an effective agent in the treatment of liver injury. Based on previous reports, the relatively short half-life (64 min) and poor bioavailability (approximately 0.31%) in rats suggested that not only sweroside itself but also its metabolites could be responsible for the observed hepato-protective effect. However, few studies have been carried out on the metabolism of sweroside. Therefore, the present study aimed at identifying the metabolites of sweroside in rat urine after a single oral dose (100 mg/kg). With ultra-high-performance liquid chromatography coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry (UHPLC/Q-TOF-MS), the metabolic profile revealed 11 metabolites in rat urine, including phase I, phase II and aglycone-related products. The chemical structures of metabolites were proposed based on accurate mass measurements of protonated or deprotonated molecules and their fragmentation patterns. Our findings showed that the aglycone of sweroside (M05) and its glucuronide conjugate (M06) were principal circulating metabolites in rats. While several other metabolic transformations, occurring via reduction, N-heterocyclization and N-acetylation after deglycosylation, were also observed. Two metabolites (M05 and M06) were isolated from the rat urine for structural elucidation and identifcation of reaction sites. Both M05 and M06 were characterized by 1H, 13C and two-dimensional nuclear magnetic resonance (NMR) spectroscopy. UHPLC/Q-TOF-MS analysis has provided an important analytical platform to gather metabolic profile of sweroside.

Occupational exposure to environmental tobacco smoke in hospitality venues: are genetic- or proteomics-based biomarkers predictive of respiratory diseases?

Environmental tobacco smoke (ETS) is recognized as an occupational hazard in the hospitality industry. Although Portuguese legislation banned smoking in most indoor public spaces, it is still allowed in some restaurants/bars, representing a potential risk to the workers’ health, particularly for chronic respiratory diseases. The aims of this work were to characterize biomarkers of early genetic effects and to disclose proteomic signatures associated to occupational exposure to ETS and with potential to predict respiratory diseases development. A detailed lifestyle survey and clinical evaluation (including spirometry) were performed in 81 workers from Lisbon restaurants. ETS exposure was assessed through the level of PM 2.5 in indoor air and the urinary level of cotinine. The plasma samples were immunodepleted and analysed by 2D-SDSPAGE followed by in-gel digestion and LC-MS/MS. DNA lesions and chromosome damage were analysed innlymphocytes and in exfoliated buccal cells from 19 cigarette smokers, 29 involuntary smokers, and 33 non-smokers not exposed to tobacco smoke. Also, the DNA repair capacity was evaluated using an ex vivo challenge comet assay with an alkylating agent (EMS). All workers were considered healthy and recorded normal lung function. Interestingly, following 2D-DIGE-MS (MALDI-TOF/TOF), 61 plasma proteins were found differentially expressed in ETS-exposed subjects, including 38 involved in metabolism, acute-phase respiratory inflammation, and immune or vascular functions. On the other hand, the involuntary smokers showed neither an increased level of DNA/chromosome damage on lymphocytes nor an increased number of micronuclei in buccal cells, when compared to non-exposed non-smokers. Noteworthy, lymphocytes challenge with EMS resulted in a significantly lower level of DNA breaks in ETS-exposed as compared to non-exposed workers (P<0.0001) suggestive of an adaptive response elicited by the previous exposure to low levels of ETS. Overall, changes in proteome may be promising early biomarkers of exposure to ETS. Likewise, alterations of the DNA repair competence observed upon ETS exposure deserves to be further understood. Work supported by Fundação Calouste Gulbenkian, ACSS and FCT/Polyannual Funding Program.

Remistura de vídeo: um estudo exploratório de diferentes estilos

Dissertação apresentada à Escola Superior de Comunicação Social como parte dos requisitos para obtenção de grau de mestre em Audiovisual e Multimédia.

Estudo comparativo da estrutura de betão armado de um edifício dimensionada segunda a actual regulamentação nacional e os eurocódigos estruturais

Dissertação para obtenção do grau de Mestre em Engenharia Civil - Ramo de Estruturas

A Review of antenas for indoor positioning systems

This paper provides a review of antennas applied for indoor positioning or localization systems. The desired requirements of those antennas when integrated in anchor nodes (reference nodes) are discussed, according to different localization techniques and their performance. The described antennas will be subdivided into the following sections according to the nature of measurements: received signal strength (RSS), time of flight (ToF), and direction of arrival (DoA). This paper intends to provide a useful guide for antenna designers who are interested in developing suitable antennas for indoor localization systems.

Proteomic analysis of the influence of the adipocyte secretome on Glioma Gl261 cells

Glioma is the most frequent form of malignant brain tumor in the adults and childhood. There is a global tendency toward a higher incidence of gliomas in highly developed and industrialized countries. Simultaneously obesity is reaching epidemic proportions in such developed countries. It has been highly accepted that obesity may play an important role in the biology of several types of cancer. We have developed an in vitro method for the understanding of the influence of obesity on glioma mouse cells (Gl261). 3T3-L1 mouse pre-adipocytes were induced to the maturity. The conditioned medium was harvested and used into the Gl261 cultures. Using two-dimension electrophoresis it was analyzed the proteome content of Gl261 in the presence of conditioned medium (CGl) and in its absence (NCGl). The differently expressed spots were collected and analyzed by means of mass spectroscopy (MALDI-TOF-MS). Significantly expression pattern changes were observed in eleven proteins and enzymes. RFC1, KIF5C, ANXA2, N-RAP, RACK1 and citrate synthase were overexpressed or only present in the CGl. Contrariwise, STI1, hnRNPs and phosphoglycerate kinase 1 were significantly underexpressed in CGl. Aldose reductase and carbonic anhydrase were expressed only in NCGl. Our results show that obesity remodels the physiological and metabolic behavior of glioma cancer cells. Also, proteins found differently expressed are implicated in several signaling pathways that control matrix remodeling, proliferation, progression, migration and invasion. In general our results support the idea that obesity may increase glioma malignancy, however, some interesting paradox finding were also reported and discussed.

Analysis of pesticide residues in strawberries and soils by GC-MS/MS, LC-MS/MS and two-dimensional GC-timeof- flight MS comparing organic and integrated pest management farming

This study analysed 22 strawberry and soil samples after their collection over the course of 2 years to compare the residue profiles from organic farming with integrated pest management practices in Portugal. For sample preparation, we used the citrate-buffered version of the quick, easy, cheap, effective, rugged, and safe (QuEChERS) method. We applied three different methods for analysis: (1) 27 pesticides were targeted using LC-MS/MS; (2) 143 were targeted using low pressure GC-tandem mass spectrometry (LP-GC-MS/MS); and (3) more than 600 pesticides were screened in a targeted and untargeted approach using comprehensive, two-dimensional gas chromatography time-of-flight mass spectrometry (GC × GC-TOF-MS). Comparison was made of the analyses using the different methods for the shared samples. The results were similar, thereby providing satisfactory confirmation of both similarly positive and negative findings. No pesticides were found in the organic-farmed samples. In samples from integrated pest management practices, nine pesticides were determined and confirmed to be present, ranging from 2 μg kg−1 for fluazifop-pbutyl to 50 μg kg−1 for fenpropathrin. Concentrations of residues in strawberries were less than European maximum residue limits.

O dever de obediência a ordens ilegais na Guarda Nacional Republicana

The Republican National Guard (GNR) is a military structure and hierarchical force where discipline and obedience is a serious matter, but at the same time, the scope of its activity relates to the protection of the rights, freedoms and guarantees of citizens and the primacy of public interest. While security force, GNR ensures democratic law, guarantee the internal security and the rights of citizens. The controversial issue that lies at the heart of this work its related with the balance between the hierarchy and the written law. The hierarchy, also established by law, with given powers, exist to apply the law. However, the rule of law has exceptions. Which institute to prioritize, hierarchy or the law. And within the law, its rules or the exceptions. Who decides? The GNR's officers have to obey the laws and regulations and comply with the accuracy and timeliness determinations, orders and instructions issued by a superior, given in terms of service, as long as does not involve the practice of crime. The GNR´s officer with command tasks exercises power of authority inherent in these functions, and the corresponding disciplinary authority, being responsible for acts by himself or by his order are practiced. Identify situations of exception to law enforcement, the situations in which one must obey illegal orders, is difficult and thankless, it requires conferred authority and raises the weight of responsibility for decisions and orders issued.

Fatores de risco no patrulhamento da GNR em ambiente operacional

In the mid-twentieth century, Portugal took the first big step towards social awareness of the Safety and Health at Work. Still later, the International Labour Organization and the World Health Organization were responsible for setting global guidelines that clarified the States for the way forward in inguito of safeguarding the common interests of workers, businesses and the state. All workers should be covered by the rules governing matters relating to Safety, imperative requirements established in the Constitution of the Portuguese Republic. These also include those soldiers from National Guard who, in contemporary social conjecture face in their everyday life situations worthy of heightened risk aquidade. Ensure the identification of risk factors to which they are exposed, is, first, a big boost in the way of preserving the safety of these employees, who daily selflessly and under the most adverse working conditions fulfill the mission of the Guarda Nacional Republicana. Adverse weather conditions, and violence at work are two examples of risk factors to which the military Guard are daily exposed, and hence arise many days of absence from the workplace. The purpose of this study is to identify the main risk factors to which the military from GNR are exposed during dismounted patrols, and also provide solutions on ways to mitigate and manage the risks presented. The cognitive distance traveled, throughout this study led us to demonstrate that it has been done by the GNR chain of Command, a huge effort to ensure through various forms (including emphasize the new Regulation of Uniforms), the resolution of the main factors that may jeopardize the integrity of the patrolmen, betting this Institution in the protection of the military that compose it, and the prevention of accidents at work through training and systematic monitoring that superiors expend with its employees.

O corpo da Guarda prisional como Órgão de Polícia Criminal e a sua importância na Segurança Interna

The study now presented a research study aimed at the exercise of security activity of the prison guard corps (CGP) in the specific context of the prison system (SP). The study also focused on the phenomenon of globalization and its influence on the current panorama of world crime and their relationship with the prison security object of study was the security activity of the CGP on the current inmate population, as well as the typology of crimes inherent to it and that requires the interpretation of the amendment to the correctional paradigm. In the preparation of the study we have tried to identify the legal framework of the profession based on the main laws and decree-laws governing the institution DGRSP and CGP's career. The difficulties and constraints were analyzed the performance of the security function of the CGP, resulting from the infrastructure, the interaction with the inmates and the need of assigning the status of Criminal Police (OPC). Really connected the relevant importance of OPC so the Corporation can establish a cooperation interaction and sharing of information with different security forces. We noted the importance of the CGP being OPC status before the framework for its action in work situations, specifically, on gatehouses, prison wings, area, and high-complexity operations such as escorts and riot control interventions.The explanation of this study enabled us to assess the importance of the actions of a police officer of the elements of the CGP for maintaining internal security. The analyses and studies were complemented by twenty years of career and exercise of the activity of the master's degree, during which time he served as guard and head of the CGP, the intervention Group and Prison Security, the Lisbon Prison, Prison, Prison of Caxias, Sintra and Monsanto.