Efeito dos inibidores das deacetilases de histonas da classe I em murganhos 3xTg-AD

Alzheimer’s disease is the commonest form of dementia worldwide. Clinically, this neurodegenerative disorder is characterized by cognitive decline, changes in personality and mood, and disorientation. Neuropathologically, it is characterized by the extracellular accumulation of beta-amyloid peptide (Aβ) forming senile plaques, and the intracellular deposition of hyperphosphorylated Tau protein forming neurofibrillary tangles. Alzheimer’s disease affects primarily the hippocampus and may progress towards the cerebral cortex. Recent studies showed that deregulation of histone acetylation is associated with increased chromatin compaction and the subsequent decrease in gene transcription. Thus, the study of inhibitors of enzymes that promote histone deacetylation, HDACs (Histone DeACetylases) became a matter of interest in Alzheimer’s disease therapeutics. Besides, it was shown that Aβ peptide induces the production of reactive oxygen species (ROS), such as hydrogen peroxide (H2O2), that further promote the activity of the Nrf2 (nuclear factor erythroid 2-related factor 2) transcription factor, altering cellular/tissue antioxidant profile. In this perspective, in this study we aimed to evaluate the possible effect of a HDAC inhibitor, sodium butyrate (SB) (1 mg/kg/day, for 42 days), in the attenuation of Alzheimer’s disease-related symptoms and neuropathology, using the triple transgenic mice for Alzheimer’s disease (3xTg-AD). 3xTg-AD mice at 10.5 months of age showed increased anxiety levels associated to the disease in comparison to non-transgenic/wild-type (WT) mice, that was not reverted by treatment with SB. Memory processes analysed through the Y-maze, the novel object recognition and object displacement tests showed no alterations in 3xTg-AD animals nor after treatment with SB. Quantification of acetylation levels of histone H3, the levels of total and phosphorylated Tau, as well as Nrf2 levels showed no significant differences between control and 3xTg-AD animals either. Treatment of 3xTg-AD mice with SB did not significantly affect any of these parameters either. Aiming to explain the lack of behavioural and biochemical differences between the 3xTg-AD and WT animals, the presence of Aβ aggregates and hyperphosphorylated Tau were assessed by immunohistochemistry in the hippocampus of 3xTg-AD mice. However, we did not observe any histopathological markers associated with this neurodegenerative disorder. The most probable explanation for these results is a delay in the disease development, leading to a later appearance of brain histopathological markers and behavioural alterations, as suggested by recent literature. In future studies, it will be interesting to analyse an animal model with a more severe pathological progression, such as the APP/PS1 transgenic mice.A doença de Alzheimer é a forma mais comum de demência a nível mundial. Clinicamente, caracteriza-se por declínio cognitivo, alterações de personalidade e humor, e desorientação. Neuropatologicamente, esta doença neurodegenerativa caracteriza-se pela acumulação extracelular do péptido beta-amilóide (βA), com formação de placas senis, e pela acumulação intracelular da proteína Tau hiperfosforilada formando tranças neurofibrilares. A patologia afeta maioritariamente o hipocampo, podendo progredir para o córtex cerebral. Estudos recentes mostram que a desregulação da acetilação das histonas está associada ao aumento da compactação da cromatina, com a consequente diminuição da transcrição génica. Assim, o estudo dos inibidores de enzimas que promovem a deacetilação das histonas, as HDAC (do inglês histone deacetylase), tem vindo a ganhar interesse como potenciais terapias contra a doença de Alzheimer. Para além disto, o péptido βA induz a produção de espécies reativas de oxigénio (ERO), tais como o peróxido de hidrogénio (H2O2), que promovem a atividade do fator de transcrição Nrf2 (do inglês nuclear factor erythroid 2-related factor 2), alterando assim a resposta antioxidante. Nesta perspetiva, neste estudo pretendemos avaliar o efeito de um inibidor das HDAC, o butirato de sódio (SB, do inglês sodium butyrate) (1 mg/kg/dia, durante 42 dias), na modificação dos sintomas e neuropatologia associados à doença de Alzheimer, utilizando o murganho triplo transgénico para a doença de Alzheimer (3xTg-AD). Os animais 3xTg-AD com 10.5 meses de idade apresentaram um nível elevado de ansiedade em comparação com animais não transgénicos (wild-type, WT), que não foi minimizado pelo tratamento com SB. Por outro lado, em testes comportamentais que permitem avaliar processos mnemónicos, como o labirinto em Y (Y-maze), o reconhecimento de novos objetos e o teste de deslocação de objetos, não se observaram alterações significativas nos animais 3xTg-AD. A determinação dos níveis de acetilação da histona H3, dos níveis da proteína Tau total e fosforilada, assim como os níveis de Nrf2 nestes animais, com recurso à técnica de Western blotting mostrou que não só não existiam diferenças entre os animais 3xTg-AD e animais controlo wild-type, como o tratamento dos animais 3xTg-AD com SB não afetou nenhum destes parâmetros. Para tentar explicar a inexistência de diferenças entre os animais 3xTg-AD e WT avaliou-se a presença de agregados de βA e de Tau hiperfosforilada por imunohistoquímica nos animais 3xTg-AD. Porém, não se verificou a presença de marcadores histopatológicos da doença nestes animais. A explicação mais provável para estes resultados poderá ser um atraso no desenvolvimento da doença causado pelo aparecimento mais tardio dos marcadores histopatológicos e das alterações comportamentais, como sugerido na literatura recente sobre este modelo transgénico. Futuramente, será necessária a realização deste estudo num modelo animal que apresente uma progressão mais acelerada da patologia, nomeadamente no murganho transgénico APP/PS1.Mestrado em Biologia Molecular e Celula

Evaluation system in the European Commission : DEVCO’S evaluation methodology

Mestrado em Economia e Políticas PúblicasNas últimas décadas, a procura relativamente a estudos de avaliação tem sido crescente e a Comissão Europeia teve um papel importante na dinamização de práticas de avaliação na União Europeia (e os seus Estados -Membros), através da institucionalização com sucesso de um sistema de avaliação e de um quadro conceptual abrangente para a avaliação de as suas actividades. Embora não exista uma metodologia única, procedimentos comuns são utilizados em várias unidades de avaliação da Comissão Europeia e algumas diretrizes foram estabelecidas entre as Direcções -Gerais que têm uma mais longa tradição em avaliação, nomeadamente a DG DEVCO. O processo de avaliação é longo e complexo, envolve muitos stakeholders e, portanto, os limites estabelecidos por uma metodologia clara e uniforme permite aferir a integridade e a independência dos resultados. No entanto, muitas vezes também provoca a perda do sentimento de pertença pela mesma, tornando mais difícil a incorporação dos resultados da avaliação no processo de tomada de decisão.Over the last decades, the demand for evaluation has been growing and the European Commission have had a major role in boosting evaluation practices in the European Union (and its Member States) by successfully institutionalising an evaluation system and establishing a comprehensive framework for the evaluation of its activities. Although no common methodology has been defined within the European Commission Directorate-Generals, common procedures are used across several evaluation units and some guidelines were established among some of them that have a longer the tradition in evaluation, notably DG DEVCO. The evaluation process is long and complex, involves many stakeholders and therefore the boundaries established by a clear and uniformed methodology enhances the integrity and independence of the results. However, it also often causes the loss of ownership, making more difficult the incorporation of the evaluation results into the decision-making process

Immunosensor for alpha-synuclein detection in Parkinson´s Disease

Parkinsons disease (PD) is a neurodegenerative disorder and one of the leading causes of death worldwide [1]. The principal neuropathological defining hallmark of PD is the presence of Lewy bodies mainly composed of aggregates of alpha-synuclein (-syn) [2]. This protein is a specific PD biomarker that, when detected early, can contribute to an effective PD treatment application. Immunosensors are analytical platforms that can detect disease biomarkers with great accuracy [3]. Thus, a selective and sensitive immunosensor to detect -syn protein based on a standard screen-printed gold electrode is proposed herein. The sensor assembly was done through three different and sequential steps: (1) incubation with 3-aminophenol (3-AMP) to promote the amination of the gold electrode surface; (2) incubation with the -syn antibody; and (3) bovine serum albumin application to avoid non-specific binding. The electrochemical characterization and surface modification was analysed through cyclic voltammetry, electrochemical impedance spectroscopy, and square wave voltammetry techniques. Finally, the performance of this immunosensor was assessed using several calibration curves with standard solutions. The final immunosensor exhibited a linear response from 7 pM to 70 nM. On balance, this immunosensor was characterized as a sensitive, selective, and reproducible platform, with great potential for real sample analysis.info:eu-repo/semantics/publishedVersio

Lideranças políticas no Brasil: da Teologia da Libertação ao Neofundamentalismo

O presente artigo tem como objetivo traçar um perfil de lideranças políticas que começaram suas trajetórias influenciadas pela Teologia da Libertação (corrente teológica que vinculava a atuação dos cristãos, principalmente católicos, a uma opção preferencial pelos pobres). Dentre estas destacamos: Chico Alencar (PSOL); Paulo Teixeira (PT); e João Pedro Stédile (MST). Também analisaremos o perfil de algumas lideranças que estão conectadas ao neofundamentalismo (movimento formado, principalmente, por evangélicos que atua na defesa de valores da moral cristã fundamentalista) e que representam, atualmente, um segmento expressivo na vida política brasileira. Dentre eles, podemos citar: João Campos (PRB), que é líder da Frente Parlamentar Evangélica e Marco Feliciano (PSC). Análise comparativa na formação das lideranças políticas é fundamental para podermos acompanhar as alterações que estão ocorrendo na composição destas correntes políticas e religiosas no Congresso Nacional

Materials, components and technology for the footwear of the future - Newalk project

At present, footwear is considered as one of the most internationalized sectors of the Portuguese industry with a significant importance for the national economy, as it is proved by the economic data of 2012: the average annual production was 74 million of shoe pares, being 90.0% of the production exported, which represents an average value of 1610 million euros [l].Financiai support from COMPETE, QREN and EU (project QREN SI J&D Copromoção 13850 NEWALK) and FCT and FEDER under Program COMPETE (PEst-C/EQB/LA0020/2011 strategic project).info:eu-repo/semantics/publishedVersio

RKIP Inhibition in Cervical Cancer Is Associated with Higher Tumor Aggressive Behavior and Resistance to Cisplatin Therapy

Cervical cancer is one of the most common cancers in women worldwide, being high-risk group the HPV infected, the leading etiological factor. The raf kinase inhibitory protein (RKIP) has been associated with tumor progression and metastasis in several human neoplasms, however its role on cervical cancer is unclear. In the present study, 259 uterine cervix tissues, including cervicitis, cervical intraepithelial lesions and carcinomas, were analyzed for RKIP expression by immunohistochemistry. We found that RKIP expression was significantly decreased during malignant progression, being highly expressed in non-neoplastic tissues (54% of the samples; 73/135), and expressed at low levels in the cervix invasive carcinomas (∼15% (19/124). Following in vitro downregulation of RKIP, we observed a viability and proliferative advantage of RKIP-inhibited cells over time, which was associated with an altered cell cycle distribution and higher colony number in a colony formation assay. An in vitro wound healing assay showed that RKIP abrogation is associated with increased migratory capability. RKIP downregulation was also associated with an increased vascularization of the tumors in vivo using a CAM assay. Furthermore, RKIP inhibition induced cervical cancer cells apoptotic resistance to cisplatin treatment. In conclusion, we described that RKIP protein is significantly depleted during the malignant progression of cervical tumors. Despite the lack of association with patient clinical outcome, we demonstrate, in vitro and in vivo, that loss of RKIP expression can be one of the factors that are behind the aggressiveness, malignant progression and chemotherapy resistance of cervical cancer.</p

HER Family Receptors are Important Theranostic Biomarkers for Cervical Cancer: Blocking Glucose Metabolism Enhances the Therapeutic Effect of HER Inhibitors

Persistent HPV infection alone is not sufficient for cervical cancer development, which requires additional molecular alterations for tumor progression and metastasis ultimately leading to a lethal disease. In this study, we performed a comprehensive analysis of HER family receptor alterations in cervical adenocarcinoma. We detected overexpression of HER protein, mainly HER2, which was an independent prognostic marker for these patients. By using in vitro and in vivo approaches, we provided evidence that HER inhibitors, allitinib and lapatinib, were effective in reducing cervical cancer aggressiveness. Furthermore, combination of these drugs with glucose uptake blockers could overcome the putative HIF1-a-mediated resistance to HER-targeted therapies. Thus, we propose that the use of HER inhibitors in association with glycolysis blockers can be a potentially effective treatment option for HER-positive cervical cancer patients.FINEP (MCTI/FINEP/MS/SCTIE/DECIT-BIOPLAT (1302/13), Brazil and co-funded by the project “ON.2 SR&TD Integrated Program (NORTE-07-0124-FEDER-000017)” co-funded by Programa Operacional Regional do Norte (ON.2- O Novo Norte), Quadro de Referência Estratégico Nacional (QREN), through Fundo Europeu de Desenvolvimento Regional (FEDER). OM is recipient of a post-doc fellowship (SFRH/BPD/108351/2015) from Fundação para a Ciência e Tecnologia (FCT), Portugal. FC is recipient of a master fellowship (2014/03684-0) from Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP). VMG is recipient from a PhD fellowship (SFRH/BD/51997/2012) from Fundação para a Ciência e Tecnologia (FCT), Portugalinfo:eu-repo/semantics/publishedVersio

Mapping cultural policy in Portugal: From incentives to crisis

Taking into account the course of cultural policy in democratic Portugal, and against the backdrop of the international crisis of 2008 and the sovereign debt crisis of 2011, this article seeks to interpret recent changes in the cultural sector in Portugal. Using both qualitative and quantitative methods it focuses on three main aspects: institutionalisation of democratic cultural policy; government funding; cultural organizations and facilities. The 2008 crisis put an end to a period in which investment tended to grow. We place Portugal in the broader European context, concluding that the Portuguese cultural scene may once again diverge from that of other European countries