RCAAP Repository

γδT cells represent the majority of lymphocytes in several mucosal tissues where they contribute to tissue homoeostasis, microbial defence and wound repair. Here we characterise a population of interleukin (IL) 17-producing γδ (γδ17) T cells that seed the testis of naive C57BL/6 mice, expand at puberty and persist throughout adulthood. We show that this population is foetal-derived and displays a T-cell receptor (TCR) repertoire highly biased towards Vγ6-containing rearrangements. These γδ17 cells were the major source of IL-17 in the testis, whereas αβ T cells mostly provided interferon (IFN)-γ in situ. Importantly, testicular γδ17 cell homoeostasis was strongly dependent on the microbiota and Toll-like receptor (TLR4)/IL-1α/IL-23 signalling. We further found that γδ17 cells contributed to tissue surveillance in a model of experimental orchitis induced by intra-testicular inoculation of Listeria monocytogenes, as Tcrδ-/- and Il17-/- infected mice displayed higher bacterial loads than wild-type (WT) controls and died 3 days after infection. Altogether, this study identified a previously unappreciated foetal-derived γδ17 cell subset that infiltrates the testis at steady state, expands upon puberty and plays a crucial role in local tissue immune surveillance.

Excitotoxicity is a phenomenon that describes the toxic actions of excitatory neurotransmitters, primarily glutamate, where the exacerbated or prolonged activation of glutamate receptors starts a cascade of neurotoxicity that ultimately leads to the loss of neuronal function and cell death. In this process, the shift between normal physiological function and excitotoxicity is largely controlled by astrocytes since they can control the levels of glutamate on the synaptic cleft. This control is achieved through glutamate clearance from the synaptic cleft and its underlying recycling through the glutamate-glutamine cycle. The molecular mechanism that triggers excitotoxicity involves alterations in glutamate and calcium metabolism, dysfunction of glutamate transporters, and malfunction of glutamate receptors, particularly N-methyl-D-aspartic acid receptors (NMDAR). On the other hand, excitotoxicity can be regarded as a consequence of other cellular phenomena, such as mitochondrial dysfunction, physical neuronal damage, and oxidative stress. Regardless, it is known that the excessive activation of NMDAR results in the sustained influx of calcium into neurons and leads to several deleterious consequences, including mitochondrial dysfunction, reactive oxygen species (ROS) overproduction, impairment of calcium buffering, the release of pro-apoptotic factors, among others, that inevitably contribute to neuronal loss. A large body of evidence implicates NMDAR-mediated excitotoxicity as a central mechanism in the pathogenesis of many neurodegenerative diseases, including amyotrophic lateral sclerosis (ALS), Alzheimer's disease (AD), and epilepsy. In this review article, we explore different causes and consequences of excitotoxicity, discuss the involvement of NMDAR-mediated excitotoxicity and its downstream effects on several neurodegenerative disorders, and identify possible strategies to study new aspects of these diseases that may lead to the discovery of new therapeutic approaches. With the understanding that excitotoxicity is a common denominator in neurodegenerative diseases and other disorders, a new perspective on therapy can be considered, where the targets are not specific symptoms, but the underlying cellular phenomena of the disease.

In Europe, which has an influenza vaccination coverage of 45·6%, about 30 million individuals have ischaemic heart disease, and more than half a million of these people will die from cardiovascular causes (figures based on the conservative standardised mortality rates of individuals aged 65 years and older). Considering that influenza vaccination is associated with a 24% risk reduction of cardiovascular mortality in this population (the lower limit of 95% CI of the estimate), an increase of coverage to 75% (as recommended by the European Council) would translate to about 42 000 potentially avoidable cardiovascular deaths in Europe among individuals with ischaemic heart disease.

Este relatório foi realizado no âmbito do Mestrado em Ensino de Geografia, da Universidade de Lisboa e apresenta o resultado de uma sequência letiva didática, através da lecionação do tema “A dinâmica do Litoral”. A sequência foi implementada na Escola Básica de Mafra à turma do 7ºA. O objetivo principal passou por perceber de que forma o estudo a dinâmica do litoral no 7º ano de escolaridade pode sensibilizar para a Educação Ambiental. Assim, é feita uma primeira abordagem à importância do Ensino da Geografia e à crescente preocupação que esta disciplina demonstra para com a proteção e prevenção do ambiente. Para conhecer a realidade portuguesa, no que diz respeito aos desafios relacionados com o litoral e com o ambiente, a estratégia passou pela exploração e análise de imagens vídeos, e da realização de algumas fichas de trabalho e questionários com o intuito de envolver os alunos na compreensão dos problemas sobre o litoral e procura de soluções para os mesmos. Para estimular o desenvolvimento de atitudes de sensibilização para a preservação do ambiente no espaço de vivência dos alunos, foram realizadas duas atividades: a primeira consistia na elaboração de um cartaz onde foram debatidas ideias sobre os problemas que afetam o mar trabalhando o pensamento critico e o pensamento criativo. A segunda atividade está relacionada com a realização de um vídeo, para concurso, com o tema “A importância do mar na minha região”, onde os alunos ficaram a perceber a importância que o mar teve e tem para o desenvolvimento de atividades locais. As estratégias de ensino – aprendizagem adotadas foram implementadas tentando sempre cativar e apelar à participação dos alunos nas aulas facilitando a aprendizagem. Estas passaram pela análise de mapas, imagens, realização fichas de trabalho e exploração e realização de vídeos. Com esta experiência foi possível concluir que o estudo da dinâmica do litoral pode de facto ajudar a sensibilizar os alunos para a Educação Ambiental. Embora estes alunos demonstrassem já alguma preocupação com as questões ambientais, tomaram ainda mais consciência dos problemas e possíveis soluções.

Formation of blood vessel networks by sprouting angiogenesis is critical for tissue growth, homeostasis and regeneration. How endothelial cells arise in adequate numbers and arrange suitably to shape functional vascular networks is poorly understood. Here we show that YAP/TAZ promote stretch-induced proliferation and rearrangements of endothelial cells whilst preventing bleeding in developing vessels. Mechanistically, YAP/TAZ increase the turnover of VE-Cadherin and the formation of junction associated intermediate lamellipodia, promoting both cell migration and barrier function maintenance. This is achieved in part by lowering BMP signalling. Consequently, the loss of YAP/TAZ in the mouse leads to stunted sprouting with local aggregation as well as scarcity of endothelial cells, branching irregularities and junction defects. Forced nuclear activity of TAZ instead drives hypersprouting and vascular hyperplasia. We propose a new model in which YAP/TAZ integrate mechanical signals with BMP signaling to maintain junctional compliance and integrity whilst balancing endothelial cell rearrangements in angiogenic vessels.

O turismo é um dos aspetos da modernidade mais importantes no mundo, com um impacto de extrema importância da economia, às sociedades e culturas, e ao ambiente. No entanto, o crescimento descontrolado do turismo tem vindo a provocar uma série de consequências no ambiente, como o aumento das emissões CO2, a sobre-exploração dos recursos naturais ou a destruição de ecossistemas e habitats naturais. O turismo sustentável e o turismo responsável são conceitos que tentam reduzir esses riscos, mas a sua diminuição deve começar pelas práticas consideradas como comuns e banais, no dia-a-dia de cada pessoa, que reduzem o consumo dos recursos e o desperdício. Considerando que o turismo jovem se encontra em franco crescimento, e que os jovens ditarão o comportamento do futuro, a presente dissertação pretende analisar as atitudes e comportamentos dos jovens face ao ambiente em férias, comparando-os com as práticas realizadas no seu quotidiano. Para tal, foram questionados 356 jovens entre os 18 e 30 anos, de várias nacionalidades, sobre os seus hábitos sustentáveis em ambos os contextos. Os resultados obtidos indicaram que os jovens revelam preocupações com o ambiente e que procuram, em geral, ter comportamentos ecologicamente corretos quer no seu dia-a-dia como em férias, não se observando grandes variações de comportamento naqueles dois contextos. Ainda assim, foram identificados alguns fatores que levam os jovens a não adotar certas práticas sustentáveis em férias, pelo que é necessária uma investigação mais profunda sobre as atitudes e motivações deste grupo-alvo, que vá além das práticas mais genéricas, a fim de criar soluções sustentáveis, credíveis e uma implementação coerente dessas práticas no quotidiano e em férias.

Este documento descreve detalhadamente todo o trabalho de estágio desenvolvido durante 9 meses na empresa GEDI (Gabinete de Estudos e Divulgação Informática, SA) pelo aluno Dan Mihai Ile, no âmbito do Mestrado em Engenharia Informática da Faculdade de Ciências da Universidade de Lisboa. Consta num projecto desenvolvido com objectivo de melhorar a qualidade, metodologia de desenvolvimento e utilização da framework SIAG tanto pelos utilizadores como pela equipa de desenvolvimento da mesma empresa. Pretende-se desenvolver um sistema que permita a criação de componentes de interface dinâmicos directamente no browser utilizando tecnologias como AJAX com interface web 2.0, GWT, web-services e sistema de mensagens para comunicação distribuída entre vários componentes da framework.

Central memory T cells (TCM) patrol lymph nodes, providing central immunosurveillance against known pathogens, but have not been described as conducting primary tissue immunosurveillance. We analyzed the expression of tissue-homing addressins in human TCM vs effector memory T cells (TEM) from the same donors. In humans, the majority of human TCM were tropic for either skin or gut, and the overall tissue tropism of TCM was comparable to that of TEM TCM were present in healthy, noninflamed human skin, lung, colon, and cervix, suggesting a role for TCM in the primary immunosurveillance of peripheral tissues. TCM also had potent effector functions; 80% of CD8+ TCM produced TC1/TC2/TC17/TC22 cytokines. TCM injected into human skin-grafted mice migrated into skin and induced inflammatory eruptions comparable to TEM-injected mice. In summary, human TCM express peripheral tissue-homing receptors at levels similar to their effector memory counterparts, are found in healthy human tissues, have impressive effector functions, and can act alone to induce skin inflammation in human engrafted mice. Our studies support a novel role for human TCM in primary immunosurveillance of peripheral tissues and highlight the important role of this long-lived cell type in tissue-based immune responses.

Campylobacter jejuni and Campylobacter coli are the most common cause of bacterial gastroenteritis worldwide. Additionally, C. jejuni is the most common bacterial etiological agent in the autoimmune Guillain-Barré syndrome (GBS). Ganglioside mimicry by C. jejuni lipooligosaccharide (LOS) is the triggering factor of the disease. LOS-associated genes involved in the synthesis and transfer of sialic acid (glycosyltranferases belonging to family GT-42) are essential in C. jejuni to synthesize ganglioside-like LOS. Despite being isolated from GBS patients, scarce genetic evidence supports C. coli role in the disease. In this study, through data mining and bioinformatics analysis, C. coli is shown to possess a larger GT-42 glycosyltransferase repertoire than C. jejuni. Although GT-42 glycosyltransferases are widely distributed in C. coli population, only a fraction of C. coli strains (1%) are very likely able to express ganglioside mimics. Even though the activity of C. coli specific GT-42 enzymes and their role in shaping the bacterial population are yet to be explored, evidence presented herein suggest that loss of function of some LOS-associated genes occurred during agriculture niche adaptation.

Micro-RNAs (miRNAs) are small non-coding RNAs that act as negative regulators of the genomic output. Their intrinsic importance within cell biology and human disease is well known. Their mechanism of action based on the base pairing binding to their cognate targets have helped the development not only of many computer applications for the prediction of miRNA target recognition but also of specific applications for functional assessment and analysis. Learning about miRNA function requires practical training in the use of specific computer and web-based applications that are complementary to wet-lab studies. In order to guide the learning process about miRNAs, we have created miRNAtools (http://mirnatools.eu), a web repository of miRNA tools and tutorials. This article compiles tools with which miRNAs and their regulatory action can be analyzed and that function to collect and organize information dispersed on the web. The miRNAtools website contains a collection of tutorials that can be used by students and tutors engaged in advanced training courses. The tutorials engage in analyses of the functions of selected miRNAs, starting with their nomenclature and genomic localization and finishing with their involvement in specific cellular functions.

Less is more: The efficacy of antibody-drug conjugates (ADCs) for cancer therapy is traditionally associated with cleavable linkers for payload release. Evidence now suggests that simpler constructs without cleavable moieties can afford more stable and effective ADCs.

Although yet poorly defined and often misused, the concept of functional mobility has been used in research studies as a more global and ecological outcome of patients’ health status. Functional mobility is a person’s physiological ability to move independently and safely in a variety of environments in order to accomplish functional activities or tasks and to participate in the activities of daily living, at home, work and in the community. Parkinson’s disease (PD) has a direct impact on patients’ motor control and on mobility in general. Even with optimal medical management, the progression of PD is associated with mounting impairments at different levels of body function, causing marked limitations in a wide variety of activities, as well as a severe disability and loss of autonomy. Despite this, for everyday functioning PD patients need to have a good functional mobility that allow them to get around effortlessly in a reasonable amount of time to access to the same environments as others. This paper reviewed the concept of functional mobility applied to PD. This was done through an International Classification of Functioning and Disability (ICF) perspective. Recommendations to address the known factors that contribute to a poor functional mobility were outlined while suggestions for clinical practice and research were made.

In the second edition of the Huntington’s Disease Clinical Trials Corner we list all currently registered and ongoing clinical trials, summarise the top-line results of the recently-announced IONIS-HTTRX trial (NCT02519036), expand on Wave Life Sciences’ PRECISION-HD1 (NCT03225833) and PRECISION-HD2 (NCT03225846), and cover one recently finished trial: the FIRST-HD deutetrabenazine trial (NCT01795859).

Four series of primaquine (PQ) derivatives were screened for antitubercular and antiplasmodial activity: amides 1a-k, ureas 2a-s, semicarbazides 3a-c and bis-ureas 4a-u. Antimycobacterial activity of PQ derivatives against Mycobacterium tuberculosis (MTB), M. avium complex (MAC) and M. avium subsp. paratuberculosis (MAP) were evaluated in vitro and compared with PQ and the standard antitubercular drugs. In general, the PQ derivatives showed higher potency than the parent compound. Most of the compounds of series 1 and 2 showed high activity against MAP, comparable or even higher than the relevant drug ciprofloxacin, and weak or no activity against MTB and MAC. bis-Trifluoromethylated cinnamamide 1k showed low cytotoxicity and high activity against all three Mycobacterium species and their activities were comparable or slightly higher than those of the reference drugs. PQ urea derivatives with hydroxyl, halogen and trifluoromethyl substituents on benzene ring 2f-p exerted very strong antimycobacterial activity towards all tested mycobacteria, stronger than PQ and the relevant standard drug(s). Unfortunately, these compounds had relatively high cytotoxicity, except bromo 2l and trifluoromethyl 2m, 2n derivatives. In general, meta-substituted derivatives were more active than analogues para-derivatives. Phenyl ureas were also more active than cycloalkyl or hydroxyalkyl ureas. Semicarbazide 3a showed similar activity as PQ, while the other two semicarbazides were inactive. Bis-urea derivatives 4 were generally less active than the urea derivatives sharing the same scaffold, differing only in the spacer type. Out of 21 evaluated bis-urea derivatives, only p-Cl/m-CF3 phenyl derivative 4p, benzhydryl derivatives 4t and 4u and bis-PQ derivative 4s showed high activity, higher than all three reference drugs. After comparison of activity and cytotoxicity, urea 2m and bis-urea 4u could be considered as the most promising agents. Antimalarial potential of PQ derivatives in vitro against the liver stage of P. berghei was evaluated as well. 3-(4-Chlorophenyl)-1-[({4-[(6-methoxyquinolin-8-yl)amino]pentyl}carbamoyl)amino]urea (4l) was the most active compound (IC50 = 42 nM; cytotoxicity/activity ratio >2000). Our results bring new insights into development of novel anti-TB and antimalarial compounds.

Genome engineering is a branch of modern biotechnology composed of a cohort of protocols designed to construct and modify a genotype with the main objective of giving rise to a desired phenotype. Conceptually, genome engineering is based on the so called genome editing technologies, a group of genetic techniques that allow either to delete or to insert genetic information in a particular genomic locus. Ten years ago, genome editing tools were limited to virus-driven integration and homologous DNA recombination. However, nowadays the uprising of programmable nucleases is rapidly changing this paradigm. There are two main families of modern tools for genome editing depending on the molecule that controls the specificity of the system and drives the editor machinery to its place of action. Enzymes such as Zn-finger and TALEN nucleases are protein-driven genome editors; while CRISPR system is a nucleic acid-guided editing system. Genome editing techniques are still not widely applied for the design of new compounds with pharmacological activity, but they are starting to be considered as promising tools for rational genome manipulation in biotechnology applications. In this review we will discuss the potential applications of programmable nucleases for the metabolic engineering of secondary metabolites with biological activity.

Introduction: Infections are a major cause of morbidity and mortality in systemic inflammatory rheumatic diseases and the management of infectious complications in patients under biological therapies deserves particular attention. Objective: Develop evidence-based recommendations for the management of infections in rheumatic patients receiving biological therapies. Methods: A search in PubMed (until 10 November 2014) and EMBASE (until 20 December 2014) databases was performed. Patients with systemic inflammatory rheumatic diseases treated with approved biologics in whom infections occurred were included. Search results were submitted to title and abstract selection, followed by detailed review of suitable studies. Information regarding presentation of the infectious complication, its diagnosis, treatment, and outcome, as well as maintenance or discontinuation of the biological agent was extracted and subsequently pooled according to the type of infection considered. Results of literature review were presented and critically reviewed in a dedicated meeting by a multidisciplinary panel. Recommendations were then formulated using the Delphi method. Finally, the level of agreement among rheumatologists was voted using an online survey. Results: Fifteen recommendations were issued. Nine general recommendations concerned the assessment of infectious risk before and while on biologics, the procedures in case of suspected infection and the management of biologics during infectious complications. Six specific recommendations were developed for respiratory, urinary, gastrointestinal, skin, osteoarticular and disseminated infections. Conclusion: These fifteen recommendations are intended to help rheumatologists in the management of infections in patients on biological therapy. They integrate an extensive literature review, expert opinion and inputs from Portuguese rheumatologists.

Introduction: Pancreas angiosarcoma is a very aggressive malignant neoplasm. The symptoms are nonspecific and it is usually diagnosed at an advanced stage, which confers a poor prognosis. Presentation of case: We present a 56-year-old woman with abdominal epigastric pain and nausea. The abdominal CT-scan showed a 7 cm mass within the head of the pancreas and the pathology and immunochemistry analysis were positive for pancreas angiosarcoma. Intra-operatively the tumor was irresectable. Discussion: Pancreas angiosarcoma is an extremely rare neoplasm with non-specific diagnosis. The histology has a wide range of presentations and immunohistochemistry is required. The surgery appears to be the only effective treatment. Conclusion: We report the seventh case of pancreas angiosarcoma in the English literature. Despite it's irresectability, the patient was asymptomatic two months after surgery, initiating chemotherapy with paclitaxel, with good tolerance.

Para mitigar os efeitos da perda e fragmentação dos habitats a rede Natura2000 tem-se tornado um importante aspecto de conservação sustentável, mas é também essencial implementar conexões funcionais entre Sítios. Os invertebrados podem actuar como importantes bioindicadores dos efeitos da fragmentação e os carabídeos são frequentemente utilizados neste contexto. As alterações climáticas podem ter um impacto significativo na biodiversidade, sendo importante perceber como as espécies vão responder a tais alterações. A modelação da distribuição potencial de espécies facilita a localização de habitats adequados e está no cerne da maioria das abordagens para a identificação de corredores ecológicos. Este estudo pretende modelar a distribuição de duas espécies de carabídeos, contribuir para um melhor conhecimento da distribuição e possíveis determinantes ecológicos das mesmas na área de estudo, analisar como os modelos de distribuição de espécies ajudam na definição de corredores ecológicos e perceber como as alterações climáticas podem afectar a distribuição das duas espécies e a persistência dos corredores ecológicos no futuro. Assim, foram recolhidos dados de presença nos Sítio Cabrela e Costa Sudoeste e na área entre ambos, para construir modelos preditivos de distribuição potencial e modelos para identificar corredores ecológicos. Os resultados destacam a importância de conhecer a distribuição de uma espécie para se poderem estimar corredores ecológicos relevantes. Só assim as medidas e esforços de gestão e conservação podem ser eficazes na preservação da conectividade. O cenário futuro B1 revelou as melhores previsões de distribuição potencial e persistência de corredores ecológicos para as duas espécies. A sobrevivência destas a longo prazo depende também da identificação de áreas que sejam importantes para a sua ocorrência e que permitam a sua dispersão. A aplicação de medidas de gestão e conservação nessas áreas é fundamental. Grândola revelou-se uma área de grande importância para a presença das espécies e para a conectividade entre os dois Sítios, funcionando ainda como um potencial refúgio para as espécies no futuro.

Esta Newsletter (NL) resulta de uma parceria entre o Instituto de Saúde Baseada na Evidência e a Cochrane Portugal, e tem como objectivo disponibilizar informação sobre áreas interessantes para a prática clínica, com base na melhor evidência científica. São incluídos estudos relevantes, criticamente avaliados pela sua validade, importância dos resultados e aplicabilidade prática, resumidos numa óptica de suporte à decisão. É dada prioridade a estudos de causalidade incluindo-se ainda, quando justificado, estudos qualitativos e metodológicos, assim como revisões científicas. O conteúdo da NL é da exclusiva responsabilidade do(s) seu(s) autor(es).

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