GDNF modulates HO-1 expression in substantia nigra postnatal cell cultures

Heme oxygenase-1 (HO-1) has been strongly highlighted because of its induction in many cell types by toxic stimuli, including oxidative stress. The intense HO-1 immunostaining in the substantia nigra of Parkinson disease (PD) patients suggests its involvement in the pathogenesis of this neurodegenerative disease. In this work we investigated HO-1 expression in rat substantia nigra postnatal cell cultures under conditions mimicking dopamine toxicity and its modulation by glial cell line-derived neurotrophic factor (GDNF), a potent neuroprotective factor for dopaminergic neurons. In neuron-glia cultures, we found that H2O2, a product of dopamine metabolism, or l-3,4-dihydroxyphenylalanine (l-DOPA), the dopamine precursor used in the therapy of PD, induced a fast up-regulation of HO-1 mRNA and protein levels, followed by a secondary down-regulation. H2O2 and l-DOPA also increased HO-1 expression in astrocyte cultures, but with a delayed time course in H2O2-treated cultures. HO-1 expression was decreased in neuron-glia cultures under conditions under which GDNF up-regulation was observed. Because exogenously applied GDNF prevented HO-1 up-regulation in cultures treated with H2O2 or l-DOPA, and antibody neutralization of GDNF prevented the secondary HO-1 down-regulation observed in neuron-glia cultures, we propose that GDNF negatively modulates HO-1 expression induced by oxidative stress. To our knowledge, this is the first report showing the modulation of HO-1 expression by GDNF.http://www.sciencedirect.com/science/article/B6T38-4GY83GS-1/1/db5c1961511769a30a115fcbfcd902c

Rodent models of Parkinson's disease: beyond the motor symptomatology

Parkinson's disease (PD) is classically characterized by motor symptoms; however, non-motor symptoms (NMS) are increasingly recognized as relevant in disease-state, given the associated alterations in mood (depression and anxiety) and cognition. Here, particularly in regards to NMS, we aimed to compare the motor, emotional and cognitive behavior of three animal models of PD that trigger dopaminergic (DAergic) degeneration on both brain hemispheres: (i) the 6-hydroxydopamine (6-OHDA, 8 or 6 µg) lesion model; (ii) the paraquat (PQ) induced model, and (iii) a genetic model based on a-synuclein overexpression (a-syn). 6-OHDA and a-syn vector were injected bilaterally in the substantia nigra pars compacta (SNpc) of adult male Wistar rats; as for PQ delivery, micro-osmotic pumps were implanted in the interscapular region. Motor deficits were observed in all models, with histological analysis of tyrosine hydroxylase positive cells in the SNpc revealing a significant loss of DAergic neurons in all animal models. In addition, the a-syn animal model also presented a reduction in exploratory activity, and the 6-OHDA and PQ animals displayed a significant increase in both depressive- and anxiety-like behavior. Interestingly, cognitive impairment (working memory) was only observed in the 6-OHDA model. Overall, these PD models are suitable for mimicking the motor symptoms associated to PD, with each encompassing other relevant NMS components of the disorder that may prove beneficial for further studies in PD.We would like to acknowledge the funds attributed by the Portuguese Foundation for Science and Technology (FCT), the PhD scholarships to E L. Campos (SFRH/BD/47311/2008) and M. M. Carvalho (SFRH/BD/51061/2010) and the Post-Doctoral fellowship to A. C. Cristovao (SFRH/BPD/69643/2010), and to Fundacao Calouste de Gulbenkian-Programme to Support Cutting Edge Research in Life Sciences and ICVS for funding this work. We want to further acknowledge Joao Cerqueira and Nadine Correia Santos for their contribution to this work

Monitorização da efetividade da vacina antigripal utilizando o método screening

Introdução: Dada a reformulação anual da composição da vacina antigripal sazonal é necessário avaliar a efetividade da vacina antigripal (EVA) todas as épocas. No presente estudo estudou-se a viabilidade da utilização do método screening na monitorização da EVA. Materiais e Métodos: O estudo incidiu nas épocas 2004/05 a 2011/12. A EVA foi determinada comparando a cobertura da vacina (CV) nos casos de síndroma gripal (SG) positivos para a gripe com a CV na população geral, através da fórmula de Orenstein. Os SG positivos foram obtidos do Programa de Vigilância da Gripe (1, 2). A CV na população geral foi estimada na amostra ECOS (3). As estimativas da EVA foram ajustadas para a idade pelo método de Farrington (4). Resultados: No período em análise a CV na população geral oscilou entre 14,3% na época 2006/07 e 19,5% na época 2009/10. As estimativas pontuais brutas da EVA, nas épocas 2004/05 e 2011/12 situaram-se abaixo dos 20%; na maioria das épocas a EVA situou-se entre 40% e 65% e na época 2007/08 a EVA foi mais elevada (~95%). Com a exceção de 2004/05 e 2011/12, os valores da EVA foram significativos. Após ajustamento, a EVA decresceu e perdeu significado estatístico. Discussão e Conclusão: O método screening permitiu estimar a EVA em todas as épocas estudadas. Observaram-se estimativas da EVA inferiores mas comparáveis com outros estudos. Como limitações, este método só permitiu o ajuste para a idade. Contudo, se o viés se mantiver constante, este método pode ser aplicado numa perspetiva de monitorização das tendências evolutivas da EVA

Monitoring influenza vaccine effectiveness using the national influenza surveillance system

Background: Due to the annual reformulation of the influenza vaccine composition estimates of the vaccine effectiveness (VE) are required every season. A portuguese influenza surveillance system is in place, and data obtained by this system may be used to evaluate VE (compared to studies specifically designed to this purpose). This study intends to evaluate the feasibility of using the national influenza surveillance system for monitoring the influenza VE (MonitorEVA). Methods: Two different study designs were implemented to estimate VE in season 2004-05 to 2011-12: Test negative design (TND) and screening method (SM). TND used a case-control approach where laboratory confirmed incident influenza like illness patients (ILI+) were compared to laboratory negative influenza ILI patients (ILI). Eligible individuals to this study consisted on individuals from all ages that consult a general practitioner or the emergency room (ER) with ILI symptoms. VE was estimated as 1-odds ratio of being vaccinated in cases versus controls adjusted for age and month of onset by logistic regression. For the SM, VE was estimated by comparing the proportion of vaccinated cases (recruited within TND) to the vaccine coverage in the source population (obtained from a sample of households, using a telephonic survey - ECOS sample), using the Orenstein formula and the Farrington method to adjust for age group. Results: Considering TND, crude point estimates of VE were under 40% in 2004/05, 2005/06 and 2011/12 season; between 50% and 70% in 2006/07, 2008/09 and 2010/11 seasons, and above 70% in the 2007/08 season. After adjustment for age group and month of onset, the VE point estimates decreased. SM estimates were in accordance to the TND ones but for the majority of the SM VE was lower than the TND. Discussion/Conclusions: Sample size and data quality are sufficient to obtain crude VE estimates with statistical significance (if VE is higher than 50%), however allowing less precise estimates. The surveillance data allowed the VE monitoring indicating if the VE was higher than 70% and less than 50%. Improvement of data quality in the surveillance program seems a potential way of improving precision and closing the gap between the two methods proposed

Li+ Influx and Binding, and Li+/Mg2+ Competition in Bovine Chromaffin Cell Suspensions as Studied by 7Li NMR and Fluorescence Spectroscopy

Li+ influx by bovine chromaffin cells, obtained from bovine adrenal medulla, was studied in intact cell suspensions using 7Li NMR spectroscopy with the shift reagent [Tm(HDOTP)]4-. The influx rate constants, ki, were determined in the absence and in the presence of two Na+ membrane transport inhibitors. The values obtained indicate that both voltage sensitive Na+ channels and (Na+/K+)-ATPase play an important role in Li+ uptake by these cells. 7Li NMR T1 and T2 relaxation times for intracellular Li+ in bovine chromaffin cells provided a T1/T2 ratio of 305, showing that Li+ is highly, immobilized due to strong binding to intracellular structures. Using fluorescence spectroscopy and the Mg2+ fluorescent probe, furaptra, the free intracellular Mg2+ concentration in the bovine chromaffin cells incubated with 15 mM LiCl was found to increase by about mM after the intracellular Li+ concentration reached a steady state. Therefore, once inside the cell, Li+ is able to displace Mg2+ from its binding sites

Uma análise lexicométrica dos documentos oficiais sobre a cooperação transfronteiriça Espanha-Portugal

No presente artigo, apresentamos uma análise do discurso produzido nos documentos oficiais sobre a Raia Ibérica (Espanha e Portugal), tomando como referência um corpus constituído por textos (conteúdos) dos diferentes programas comunitários do INTERREG e suas diretrizes para os programas operativos, Portugal e Espanha. Para apoiar esta análise, recorremos ao tratamento lexical dos textos através do software Iramuteq, cujos resultados permitiram colocar em evidência, os núcleos de sentido que sustentam as estratégias discursivas utilizadas nos documentos políticos para a maximização da importância do sistema ideológico da cooperação transfronteiriça. Os principais resultados da análise lexicométrica permitiram colocar em evidência dois pólos do sistema ideológico da cooperação transfronteiriça Espanha-Portugal, em geral, e da Euroregião EuroACE, em particular, sendo um marcado pela visão duma utopia generosa dum mundo “sem fronteiras”, e por outro lado, a presença duma lógica ideológica local e regionalista fundada pela dinamização de jogos de poder dos atores coletivos, que, em última instância, converte a ideologia da cooperação transfronteiriça num veículo para instrumentalizar o acesso a mecanismos financeiros ou institucionais. Em suma, falar de cooperação transfronteiriça não esvazia a noção de fronteira estatal e espacial formalmente definida, antes porém, reforça a omnipresença da ideia de uma fronteira dependente de lógicas exógenas, com impactos de ordem diversa no campo social, cultural e político

Hidden lives and lost paths: (dis)connections between public policies and institutional practices on domestic violence

[PT] Este ensaio tem como finalidade conhecer as práticas organizacionais que potenciem uma intervenção mais eficaz no combate à violência doméstica, na perspetiva de vítimas e de profissionais. Trata-se de uma investigação qualitativa, baseada na análise de conteúdo a relatórios públicos e a entrevistas aplicadas a diferentes atores. O quadro legal português encontra-se atualizado e, na redação atual, assume um papel inovador no combate à violência doméstica. As práticas dos atores da Rede Nacional de Apoio às Vítimas de Violência Doméstica (RNAVVD) são dissemelhantes, encontrando-se práxis com diversos graus de sucesso. Em prol da prevenção dos homicídios em violência doméstica, recomenda-se que o Governo português promova ferramentas de monitorização da qualidade dos serviços disponibilizados às utentes vítimas de violência doméstica.[EN] This essay aims to understand the organizational practices that enhance the effectiveness of interventions in the fight against domestic violence, from the perspective of victims and professionals. This is a qualitative investigation, based on content analysis encountered in public reports and interviews applied to different actors. The Portuguese legal framework is updated and, in the current wording, assumes an innovative role on the combat of domestic violence. The practices of the actors of the National Network for The Support of Victims of Domestic Violence are dissimilar, presenting praxis with varying degrees of success. In order to prevent homicides linked to domestic violence, it is recommended that the Portuguese government promotes tools to monitor the quality of the services available to victims of domestic violence

Influenza seroprotection correlates with predominant circulating viruses during 2014/15 and 2015/16 seasons in Portugal

Rede Portuguesa de Laboratórios para o Diagnóstico da GripeBACKGROUND: Population immune profile for influenza is highly affected by circulating influenza viruses, thus changing the risk of infection for influenza. This study aims to assess influenza immunity in the Portuguese population by age groups, during 2014 and 2015 and establish a relationship between seroprotection and circulating influenza viruses in 2014/15 and 2015/16 seasons. METHODS: Two cross-sectional studies were developed based on a convenience serum sample collected in June 2014 (n=626) and July 2015 (n=675) in hospitals from mainland and Azores and Madeira.Serums equally represent all age groups. Antibody titers were evaluated by HI assay for strains recommended for seasonal influenza vaccine northern hemisphere,2014/15 and 2015/2016. Seroprevalences were estimated for each strain by age group and the association with seasonal cumulative influenza-like illness (ILI) rates for influenza virus during both seasons was analised. RESULTS: In June 2014 the highest seroprotection was observed for influenza A(H3) (39.0%; 95% CI: 36.2-43.8%) and A(H1)pdm09 (29.7; 95% CI: 26.3-33.4%), with higher levels in children 5-14 years old. In 2014/2015 a dominant circulation of influenza B/Yamagata was observed with high incidence rates in individuals under 65 years old, the ones that had lower seroprotection. Although before the start of the season high protection for A(H3) was observed, the circulation of the new drift A(H3) strains had gained an immunological advantage,in accordance with A(H3) elevated incidence rates observed during 2014/15. In July 2015 the highest seroprotection was observed for influenza B/ Yamagata (55.1%; 95% CI: 51.4-58.9%), 2.4 times the estimated 2014.This increase was even more pronounced in younger (≤ 4 years old), 6.3 times increase in 2015.This fact is in agreement with the predominant influenza B virus detected and the high ILI incidence rate observed in children during 2014/2015 epidemic. Seroprotection levels for influenza A in July 2015 were not significantly different from 2014.During 2015/16 season, influenza A(H1N1)pdm09 was predominant, with high incidence rate in < 65 year old. Influenza B/Victoria lineage,although detected at low levels increased in frequency, in agreement with the lowest level of seroprotection detected in the general population before the start of 2015/2016 season (21.8%; 95% CI: 18.7-24.0%). CONCLUSIONS There was a correlation between virus circulation, incidence rates for each age group and the previous seroprotection for seasonal influenza viruses.Our study highlights the value of measuring the serological profile for influenza to establishe risk groups for infection for which an increase preventive measures, including vaccination, should be fostered.info:eu-repo/semantics/publishedVersio

SARS-CoV-2 introductions and early dynamics of the epidemic in Portugal

Genomic surveillance of SARS-CoV-2 in Portugal was rapidly implemented by the National Institute of Health in the early stages of the COVID-19 epidemic, in collaboration with more than 50 laboratories distributed nationwide. Methods By applying recent phylodynamic models that allow integration of individual-based travel history, we reconstructed and characterized the spatio-temporal dynamics of SARSCoV-2 introductions and early dissemination in Portugal. Results We detected at least 277 independent SARS-CoV-2 introductions, mostly from European countries (namely the United Kingdom, Spain, France, Italy, and Switzerland), which were consistent with the countries with the highest connectivity with Portugal. Although most introductions were estimated to have occurred during early March 2020, it is likely that SARS-CoV-2 was silently circulating in Portugal throughout February, before the first cases were confirmed. Conclusions Here we conclude that the earlier implementation of measures could have minimized the number of introductions and subsequent virus expansion in Portugal. This study lays the foundation for genomic epidemiology of SARS-CoV-2 in Portugal, and highlights the need for systematic and geographically-representative genomic surveillance.We gratefully acknowledge to Sara Hill and Nuno Faria (University of Oxford) and Joshua Quick and Nick Loman (University of Birmingham) for kindly providing us with the initial sets of Artic Network primers for NGS; Rafael Mamede (MRamirez team, IMM, Lisbon) for developing and sharing a bioinformatics script for sequence curation (https://github.com/rfm-targa/BioinfUtils); Philippe Lemey (KU Leuven) for providing guidance on the implementation of the phylodynamic models; Joshua L. Cherry (National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health) for providing guidance with the subsampling strategies; and all authors, originating and submitting laboratories who have contributed genome data on GISAID (https://www.gisaid.org/) on which part of this research is based. The opinions expressed in this article are those of the authors and do not reflect the view of the National Institutes of Health, the Department of Health and Human Services, or the United States government. This study is co-funded by Fundação para a Ciência e Tecnologia and Agência de Investigação Clínica e Inovação Biomédica (234_596874175) on behalf of the Research 4 COVID-19 call. Some infrastructural resources used in this study come from the GenomePT project (POCI-01-0145-FEDER-022184), supported by COMPETE 2020 - Operational Programme for Competitiveness and Internationalisation (POCI), Lisboa Portugal Regional Operational Programme (Lisboa2020), Algarve Portugal Regional Operational Programme (CRESC Algarve2020), under the PORTUGAL 2020 Partnership Agreement, through the European Regional Development Fund (ERDF), and by Fundação para a Ciência e a Tecnologia (FCT).info:eu-repo/semantics/publishedVersio