Mutilação Genital Feminina: o que Sabem os Profissionais de Saúde?

Portugal is considered by the World Health Organization (WHO) a risk country for the practice of Female Genital Mutilation (FGM). Objectives: To evaluate the knowledge that health professionals from Maternity Dr. Alfredo da Costa (MAC) have regarding FGM. Population and Methods: Analysis of surveys delivered to health professionals from MAC (a hospital dedicated to reproductive health), between April and June 2008, addressing issues related to the knowledge about FGM. Results: Authors collected 112 valid surveys involving 38 doctors, 48 nurses and 26 medical auxiliaries/administrative personnel. From the respondents, 106 (95%) had heard about FMG practice before, the media being the most reported source of information; 59 (53%) replied they could be able to recognize FGM cases in their clinical practice; however, only 31 (28%) claimed to know the FGM type classiication and 32 (29%) admitted to be prepared to recognize and manage these situations in their own clinical practice; 9 had been consulted explicitly by a FGM practice complication and 1 doctor had admitted having been asked to perform/execute FGM; 13 (12%) recognized that the Portuguese legislation its this practice. Regarding the practice of FGM, 100 (89%) of respondentes stated that it should not be maintained and 97 (87%) stated that it should not be tolerated. However, 42 (38%) considered that if these practices were a reality, then they should be medical assisted. Discussion: Health professionals can play an important role in eliminating the practice of FGM, not only by the proper clinical management of this situation, but also by preventing those communities at risk to resort to FGM. Most health professionals are not prepared to deal with FGM in their clinical practice. It is important to promote a better knowledge on the subject and to create protocols for proper clinical management.

Deletions within COL11A1 in Type 2 Stickler Syndrome Detected by Multiplex Ligation - Syndrome Detected by Multiplex Ligation Dependent Probe Amplification (MLPA)

Background: COL11A1 is a large complex gene around 250 kb in length and consisting of 68 exons. Pathogenic mutations in the gene can result in Stickler syndrome, Marshall syndrome or Fibrochondrogenesis. Many of the mutations resulting in either Stickler or Marshall syndrome alter splice sites and result in exon skipping, which because of the exon structure of collagen genes usually leaves the message in-frame. The mutant protein then exerts a dominant negative effect as it co-assembles with other collagen gene products. To date only one large deletion of 40 kb in the COL11A1, which was detected by RT-PCR, has been characterized. However, commonly used screening protocols, utilizing genomic amplification and exon sequencing, are unlikely to detect such large deletions. Consequently the frequency of this type of mutation is unknown. Case presentations: We have used Multiplex Ligation-Dependent Probe Amplification (MLPA) in conjunction with exon amplification and sequencing, to analyze patients with clinical features of Stickler syndrome, and have detected six novel deletions that were not found by exon sequencing alone. Conclusion: Exon deletions appear to represent a significant proportion of type 2 Stickler syndrome. This observation was previously unknown and so diagnostic screening of COL11A1 should include assays capable of detecting both large and small deletions, in addition to exon sequencing.

Gestão de Redes e Sistemas: Estudo de um caso

A gestão e monitorização de redes é uma necessidade fundamental em qualquer organização, quer seja grande ou pequena. A sua importância tem de ser refletida na eficiência e no aumento de informação útil disponível, contribuindo para uma maior eficácia na realização das tarefas em ambientes tecnologicamente avançados, com elevadas necessidades de desempenho e disponibilidade dos recursos dessa tecnologia. Para alcançar estes objetivos é fundamental possuir as ferramentas de gestão de redes adequadas. Nomeadamente ferramentas de monitorização. A classificação de tráfego também se revela fundamental para garantir a qualidade das comunicações e prevenir ataques indesejados aumentando assim a segurança nas comunicações. Paralelamente, principalmente em organizações grandes, é relevante a inventariação dos equipamentos utilizados numa rede. Neste trabalho pretende-se implementar e colocar em funcionamento um sistema autónomo de monitorização, classificação de protocolos e realização de inventários. Todas estas ferramentas têm como objetivo apoiar os administradores e técnicos de sistemas informáticos. Os estudos das aplicações que melhor se adequam à realidade da organização culminaram num acréscimo de conhecimento e aprendizagem que irão contribuir para um melhor desempenho da rede em que o principal beneficiário será o cidadão.

Hypermedia-based Web Services as System Integrators

As we move more closely to the practical concept of the Internet of Things and, our reliance on public and private APIs increases, web services and their related topics have become utterly crucial to the informatics community. However, the question about which style of web services would best solve a particular problem, can raise signi cant and multifarious debates. There can be found two implementation styles that highlight themselves: the RPC-oriented style represented by the SOAP protocol’s implementations and the hypermedia style, which is represented by the REST architectural style’s implementations. As we search examples of already established web services, we can nd a handful of robust and reliable public and private SOAP APIs, nevertheless, it seems that RESTful services are gaining popularity in the enterprise community. For the current generation of developers that work on informatics solutions, REST seems to represent a fundamental and straightforward alternative and even, a more deep-rooted approach than SOAP. But are they comparable? Do both approaches have each speci c best suitable scenarios? Such study is brie y carried out in the present document’s chapters, starting with the respective background study, following an analysis of the hypermedia approach and an instantiation of its architecture, in a particular case study applied in a BPM context.

Estudo de vulnerabilidades em aplicações web e o seu reflexo em domínios Portugueses

Muito se tem falado sobre revolução tecnológica e do aparecimento constante de novas aplicações Web, com novas funcionalidades que visam facilitar o trabalho dos utilizadores. Mas será que estas aplicações garantem que os dados transmitidos são tratados e enviados por canais seguros (protocolos)? Que garantias é que o utilizador tem que mesmo que a aplicação utilize um canal, que prevê a privacidade e integridade de dados, esta não apresente alguma vulnerabilidade pondo em causa a informação sensível do utilizador? Software que não foi devidamente testado, aliado à falta de sensibilização por parte dos responsáveis pelo desenvolvimento de software para questões de segurança, levam ao aumento de vulnerabilidades e assim exponenciam o número de potenciais vítimas. Isto aliado ao efeito de desinibição que o sentimento de invisibilidade pode provocar, conduz ao facilitismo e consequentemente ao aumento do número de vítimas alvos de ataques informáticos. O utilizador, por vezes, não sabe muito bem do que se deve proteger, pois a confiança que depõem no software não pressupõem que os seus dados estejam em risco. Neste contexto foram recolhidos dados históricos relativos a vulnerabilidades nos protocolos SSL/TLS, para perceber o impacto que as mesmas apresentam e avaliar o grau de risco. Para além disso, foram avaliados um número significativo de domínios portugueses para perceber se os mesmos têm uma vulnerabilidade específica do protocolo SSL/TLS.

Expressão em Escherichia coli de antigénios do Cell fusing agent virus (Flaviviridae: Flavivirus) como proteína de fusão

RESUMO: O Cell Fusing Agent Vírus (CFAV), considerado como o primeiro “flavivírus específicos de insectos” (ISF), parece estar exclusivamente adaptado aos seus hospedeiros, não replicando em células de vertebrados. Apesar de ter sido identificado há mais de três décadas (1975), a verdade é que muito pouco se conhece sobre a sua biologia. Dado o seu parentesco filogenético com alguns outros flavivírus encontrados naturalmente em mosquitos de diferentes géneros colhidos em diferentes regiões do globo, este vírus poderá ser usado como modelo para o estudo de ISF. No entanto, necessitam do desenvolvimento de ferramentas básicas, tais como clones moleculares ou baterias de soros contendo anticorpos que reconheçam uma ou mais proteínas codificadas pelo genoma viral, produzidas, por exemplo, a partir de antigénios virais produzidos de forma recombinante. Com este trabalho pretendeu-se a optimização de protocolos que permitiram a expressão e purificação parcial de quatro proteínas [duas proteínas estruturais (C e E) e duas não estruturais (NS3hel e NS5B)] do CFAV em E. coli, todas elas produzidas como proteínas de fusão com “caudas” (tags) de hexahistidina nos seus extremos carboxilo. Para a expansão do CFAV foram utilizadas células Aedes albopictus (C6/36). Após a realização da extracção do RNA viral e a obtenção de cDNA, procedeu-se amplificação, por RT-PCR, das regiões codificantes das proteínas C, E, NS3hel e NS5B, utilizando primers específicos. Os quatro fragmentos de DNA foram independentemente inseridos no vector pJTE1.2/blunt usando E. coli NovaBlue como hospedeira de clonagem e, posteriormente, inseridos em vectores de expressão pET-28b e pET-29b usando E. coli BL21(DE3)pLysS e Rosetta(DE3)pLysS como hospedeiras de expressão. Após da indução, expressão e purificação das proteínas recombinantes C, E, NS3hel e NS5B, foi confirmada a autenticidade destas proteínas produzidas através do método Western Blot com um anticorpo anti-histidina. --------- ABSTRACT: The Cell Fusing Agent virus (CFAV) considered as the first "insect- specific flavivirus" (ISF) and seems to be uniquely adapted to their hosts, not replicating in vertebrate cells. Although it has been known for more than three decades (1975), the truth is very little is known about its biology. Given its close phylogenetic relationship with other flavivirus naturally circulating in various genera of mosquitoes collected from different regions of the globe, this virus could be used as a model for the study of ISF. However, such studies require the development of experimental basic tools, such as molecular clones or serum batteries containing antibodies that recognize one or more proteins encoded by the viral genome, produced, for example, from viral antigens recombinant produced. In this work, we carried out the optimization of protocols that allowed the expression and partial purification of four proteins [two structural proteins (C and E) and two nonstructural proteins (NS3hel and NS5B)] CFAV in E. coli as fusion protein for c-terminal hexahistidine tags. For the expansion of the CFAV we used Aedes albopictus (C6/36) cells. After completion of the viral RNA extraction and cDNA obtained, amplification of the coding regions of the C, E, NS5B and NS3hel proteins was carried out by RT-PCR using specific primers. The four DNA fragments were independently inserted into the vector pJTE1.2/blunt using E. coli NovaBlue as cloning host and then inserted into expression vectors pET-28b and pET-29b using E. coli BL21(DE3)pLysS and Rosetta(DE3)pLysS as expression host. After induction, expression and purification of recombinant C, E, NS3hel and NS5B proteins Western Blot analyses with an anti-histidine antibody confirmed the authenticity of these proteins produced.

Quantum dots: synthesis, functionalization and bioconjugation for biological applications

Dissertation presented to obtain the Ph.D degree in Engineering Sciences and Technology

Locally Advanced and Metastatic Prostate Cancer Treated with Intermittent Androgen Monotherapy or Maximal Androgen Blockade: Results from a Randomised Phase 3 Study by the South European Uroncological Group

BACKGROUND: Few randomised studies have compared antiandrogen intermittent hormonal therapy (IHT) with continuous maximal androgen blockade (MAB) therapy for advanced prostate cancer (PCa). OBJECTIVE: To determine whether overall survival (OS) on IHT (cyproterone acetate; CPA) is noninferior to OS on continuous MAB. DESIGN, SETTING, AND PARTICIPANTS: This phase 3 randomised trial compared IHT and continuous MAB in patients with locally advanced or metastatic PCa. INTERVENTION: During induction, patients received CPA 200 mg/d for 2 wk and then monthly depot injections of a luteinising hormone-releasing hormone (LHRH; triptoreline 11.25 mg) analogue plus CPA 200 mg/d. Patients whose prostate-specific antigen (PSA) was <4 ng/ml after 3 mo of induction treatment were randomised to the IHT arm (stopped treatment and restarted on CPA 300 mg/d monotherapy if PSA rose to ≥20 ng/ml or they were symptomatic) or the continuous arm (CPA 200 mg/d plus monthly LHRH analogue). OUTCOME MEASUREMENTS AND STATISTICAL ANALYSIS: Primary outcome measurement was OS. Secondary outcomes included cause-specific survival, time to subjective or objective progression, and quality of life. Time off therapy in the intermittent arm was recorded. RESULTS AND LIMITATIONS: We recruited 1045 patients, of which 918 responded to induction therapy and were randomised (462 to IHT and 456 to continuous MAB). OS was similar between groups (p=0.25), and noninferiority of IHT was demonstrated (hazard ratio [HR]: 0.90; 95% confidence interval [CI], 0.76-1.07). There was a trend for an interaction between PSA and treatment (p=0.05), favouring IHT over continuous therapy in patients with PSA ≤1 ng/ml (HR: 0.79; 95% CI, 0.61-1.02). Men treated with IHT reported better sexual function. Among the 462 patients on IHT, 50% and 28% of patients were off therapy for ≥2.5 yr or >5 yr, respectively, after randomisation. The main limitation is that the length of time for the trial to mature means that other therapies are now available. A second limitation is that T3 patients may now profit from watchful waiting instead of androgen-deprivation therapy. CONCLUSIONS: Noninferiority of IHT in terms of survival and its association with better sexual activity than continuous therapy suggest that IHT should be considered for use in routine clinical practice.

Multipath inter-domain policy routing

Dissertação submetida para a obtenção do grau de Doutor em Engenharia Electrotécnica e de Computadores

On the cloud deployment of a session abstraction for service/data aggregation

Dissertação para obtenção do Grau de Mestre em Engenharia Informática

Functional brain perfusion evaluation with Arterial Spin Labeling at 3 Tesla

Dissertation submitted in Faculdade de Ciências e Tecnologia of Universidade Nova de Lisboa for the degree of Master of Biomedical Engineering

Multipath policy routing in packet switched networks

Dissertação apresentada para obtenção do Grau de Mestre em Engenharia Electrotécnica e de Computadores, pela Universidade Nova de Lisboa, Faculdade de Ciências e Tecnologia

Intrusion tolerant routing with data consensus in wireless sensor networks

Dissertação para obtenção do Grau de Mestre em Engenharia Informática

Atomic force microscopy assisted with electron and ion beam microscopy for the direct measurement of biostructures and DNA samples

Dissertation to obtain a Master degree in Biotechnology