Drug-induced stress granule formation protects sensory hair cells in mouse cochlear explants during ototoxicity

Stress granules regulate RNA translation during cellular stress, a mechanism that is generally presumed to be protective, since stress granule dysregulation caused by mutation or ageing is associated with neurodegenerative disease. Here, we investigate whether pharmacological manipulation of the stress granule pathway in the auditory organ, the cochlea, affects the survival of sensory hair cells during aminoglycoside ototoxicity, a common cause of acquired hearing loss. We show that hydroxamate (-)-9, a silvestrol analogue that inhibits eIF4A, induces stress granule formation in both an auditory cell line and ex-vivo cochlear cultures and that it prevents ototoxin-induced hair-cell death. In contrast, preventing stress granule formation using the small molecule inhibitor ISRIB increases hair-cell death. Furthermore, we provide the first evidence of stress granule formation in mammalian hair cells in-vivo triggered by aminoglycoside treatment. Our results demonstrate that pharmacological induction of stress granules enhances cell survival in native-tissue, in a clinically-relevant context. This establishes stress granules as a viable therapeutic target not only for hearing loss but also other neurodegenerative diseases.EI:595 - Action on Hearing Loss; 091092/Z/09/Z - Wellcome Trust (Wellcome); MR/N004329/1 - RCUK | Medical Research Council (MRC)Published versio

Human tendon-derived cell sheets created by magnetic force-based tissue engineering hold tenogenic and immunomodulatory potential

Cell sheet technology and magnetic based tissue engineering hold the potential to become instrumen- tal in developing magnetically responsive living tissues analogues that can be potentially used both for modeling and therapeutical purposes. Cell sheet constructions more closely recreate physiological niches, through the preservation of contiguous cells and cell-ECM interactions, which assist the cellular guidance in regenerative processes. We herein propose to use magnetically assisted cell sheets (magCSs) constructed with human tendon- derived cells (hTDCs) and magnetic nanoparticles to study inflammation activity upon magCSs exposure to IL-1 β, anticipating its added value for tendon disease modeling. Our results show that IL-1 βinduces an inflammatory profile in magCSs, supporting its in vitro use to en- lighten inflammation mediated events in tendon cells. Moreover, the response of magCSs to IL-1 βis mod- ulated by pulsed electromagnetic field (PEMF) stimulation, favoring the expression of anti-inflammatory genes, which seems to be associated to MAPK(ERK1/2) pathway. The anti-inflammatory response to PEMF together with the immunomodulatory potential of magCSs opens new perspectives for their applicability on tendon regeneration that goes beyond advanced cell based modeling.This research was funded by the ERC CoG MagTendon (No. 772817), Fundação para a Ciência e Tecnologia (FCT) for the doctoral grant PD/BD/128089/2016 of A. Vinhas and the project MagTT PTDC/CTM-CTM/29930/2017 (POCI-01-0145-FEDER-29930), project Norte-01-0145-FEDER-02219015 supported by Norte Portugal Regional Operational Programme (NORTE 2020) and EC Twinning project Achilles (No. 810850)

Effect of the exposure to maternal smoking during pregnancy and childhood on the body mass index until adolescence

OBJETIVO Analisar se a exposição ao tabagismo materno durante a gravidez e no início da infância afeta as mudanças no índice de massa corporal entre o nascimento e a adolescência. MÉTODOS Realizado estudo de coorte de base populacional com 2.405 crianças (0 a 5 anos) nascidas em Cuiabá, Brasil, e avaliadas de 1999 a 2000. De 2009 a 2011, esse grupo foi reavaliado. Peso ao nascer foi obtido a partir de registros médicos e a exposição ao tabagismo durante a gravidez e infância foi avaliada na primeira entrevista. Modelos lineares de efeitos mistos foram utilizados para estimar a associação entre a exposição ao tabagismo materno, durante a gravidez e a fase pré-escolar, e o índice de massa corporal das crianças ao nascer e durante a infância e adolescência. RESULTADOS Apenas 11,3% das mães relataram fumar durante a gravidez, sendo que a maioria delas (78,2%) também fumou durante a fase pré-escolar da criança. Entre as mães que fumaram exclusivamente durante a gravidez (n = 59), 97,7% fumaram somente no primeiro trimestre. As mudanças de índice de massa corporal entre o nascimento e a infância foram semelhantes entre as crianças expostas e não expostas ao tabagismo materno. Entretanto, entre a infância e a adolescência, a taxa de variação do índice de massa corporal foi maior entre os expostos ao tabagismo materno apenas durante a gravidez quando comparado aos não expostos. CONCLUSÕES A exposição ao fumo apenas durante a gravidez, especialmente no primeiro trimestre, pode afetar as mudanças no índice de massa corporal até a adolescência, apoiando a recomendação de cessação do tabagismo entre as mulheres em idade fértil.OBJECTIVE: Investigate the effect of exposure to smoking during pregnancy and early childhood on changes in the body mass index (BMI) from birth to adolescence. METHODS: A population-based cohort of children (0-5 years old) from Cuiabá, Midwest Brazil, was assessed in 1999-2000 (n = 2,405). Between 2009 and 2011, the cohort was re-evaluated. Information about birth weight was obtained from medical records, and exposure to smoking during pregnancy and childhood was assessed at the first interview. Linear mixed effects models were used to estimate the association between exposure to maternal smoking during pregnancy and preschool age, and the body mass index of children at birth, childhood and adolescence. RESULTS: Only 11.3% of the mothers reported smoking during pregnancy, but most of them (78.2%) also smoked during early childhood. Among mothers who smoked only during pregnancy (n = 59), 97.7% had smoked only in the first trimester. The changes in body mass index at birth and in childhood were similar for children exposed and those not exposed to maternal smoking. However, from childhood to adolescence the rate of change in the body mass index was higher among those exposed only during pregnancy than among those who were not exposed. CONCLUSIONS: Exposure to smoking only during pregnancy, especially in the first trimester, seems to affect changes in the body mass index until adolescence, supporting guidelines that recommend women of childbearing age to stop smokin

Monitoring of emerging micropollutants in hydric media in Bragança district

Emerging Micropollutants are a subclass of Pollutants and they can be classified as Persistent Organic Pollutants, Endocrine Disrupting Compounds and Pharmaceuticals and Personal Care Products (PPCP´s). These compounds may cause undesired effects to health or to the environment when used in high quantities or wrong ways. Generally, a pharmaceutical drug is prescribed to cause the desired therapeutic effect. However after their use, even at low concentrations they can retain their physicochemical properties [1]. Because of these effects, among others, PPCP’s are substances excreted in relevant quantities by the human body and are not effectively removed from water in sewage treatment plants [2].info:eu-repo/semantics/publishedVersio

Doença de Pompe: a propósito de 2 casos clínicos do Hospital Pêro da Covilhã

A Doença de Pompe também conhecida como doença de armazenamento do glicogénio tipo II ou deficiência de maltase ácida, pertence a um grupo de doenças lisossomais de sobrecarga, sendo esta a forma mais grave do grupo. Trata-se de uma doença hereditária autossómica recessiva causada por mutações no gene que codifica a α-glicosidase ácida, determinando acumulação lisossomal de glicogénio. A doença de Pompe apresenta-se com um amplo espectro de manifestações clínicas e os sintomas iniciais podem surgir em qualquer idade. Previamente à aprovação, no ano 2006, da terapêutica de reposição enzimática com α-glicosidase ácida recombinante, não existia nenhum tratamento específico para a doença de Pompe. A terapêutica de reposição enzimática revelou-se eficaz e capaz de alterar a história natural da doença, dando uma nova esperança a estes doentes. Este trabalho tem como objetivo rever o estado da arte da Doença de Pompe em Portugal, com especial ênfase à forma tardia desta doença. São abordadas as formas de apresentação, patogénese, métodos de diagnósticos, bem como as atuais recomendações terapêuticas. Para tal é levada a cabo uma revisão da literatura científica mais recentemente publicada e relevante, acerca da doença em estudo. Este trabalho contempla ainda a apresentação e discussão de dois casos clínicos de doentes com a forma tardia da doença de Pompe, acompanhados no Hospital Pêro da Covilhã (Centro Hospitalar Cova da Beira).Pompe disease, also known as glycogen storage disease type II or acid maltase deficiency, belongs to a group of lysossomal storage disorders, this being the most severe form of the group. It is an autossomal recessive disease caused by mutations in the gene encoding the lysosomal enzyme acid α-glucosidase, leading to lysosomal glygogen storage. Pompe’s disease has a broad spectrum of clinical manifestations, whose initial symptoms can appear at any age. There was no disease-specific treatment for Pompe disease until the approval, in 2006, of the enzyme replacement therapy with recombinant human acid α-glucosidase. This enzyme replacement therapy is effective in modifying the natural course of the disease and gave a new hope to Pompe patients. The present work intends to review the state of the art of Pompe disease in Portugal, with particular emphasis on the late-onset disease. It is discussed the clinical presentation, pathogenesis, diagnosis methods and current treatment recommendations of this disease. In order to achieve this, a review of the latest published and relevant scientific literature is performed. This paper also includes the presentation and discussion of two patients with lateonset Pompe disease, followed at Hospital Pêro da Covilhã (Centro Hospitalar Cova da Beira)

Memórias e cinema pelo olhar da criança de periferia

Este artigo é um recorte da pesquisa “O Cinema como mediador de aprendizagem de crianças e adolescentes em situação de vulnerabilidade social", sob a orientação da Profa. Dra. Marlene Gonçalves e integrante do mestrado em Educação da Universidade Federal de Mato Grosso, Brasil. O objetivo da pesquisa é investigar a memória familiar e escolar de 30 crianças de 09 a 13 anos, relacionada aos filmes exibidos em sessões refletidas, articulando experiência, lembranças, narrativas e tecnologia, observando os impactos da vulnerabilidade social na elaboração do conhecimento. Os sujeitos são estudantes da rede pública de ensino e residem na região periférica de Várzea Grande, que ocupa a 920ª posição da lista de 5.565 municípios brasileiros segundo o Índice de Desenvolvimento Humano Municipal (IDHM), enquanto a capital Cuiabá, localizada ao lado, ocupa a 92ª, segundo ranking no "Atlas do Desenvolvimento Humano no Brasil 2013", fruto de estudos do Instituto de Pesquisa Econômica Aplicada (Ipea), do Programa das Nações Unidas para o Desenvolvimento (Pnud) e da Fundação João Pinheiro. O cenário dos sujeitos de pesquisa está inserido no estado apontado como maior produtor de soja, de algodão em pluma e rebanho bovino, em que o agronegócio representa 50,5% do PIB, segundo o Instituto Mato-Grossense de Economia Agropecuária (Imea). Apesar do potencial econômico que perpassa pela mídia brasileira quando Mato Grosso é notícia, o ambiente de pesquisa revela as contradições persistentes neste país, em que crianças de periferia vivem situações de ausências diversas, da doméstica às políticas públicas. Neste artigo, focalizamos alguns personagens reais, residentes do bairro Eliane Gomes, incrustado neste Brasil central, cujo cerrado evidencia mais que o retorcido das árvores do quintal, mas sobretudo, o contorno árduo de histórias de vidas cujas memórias estão se construindo nas salas de aulas, para além dos muros da escola e na adversidade do abandono dos pais pelo vício e pela criminalidade. Desprovidas de espaços públicos para fruição cultural, os sujeitos foram convidados a percorrer novo trajeto, em que a filmografia foi o fio condutor. Nesta caminhada, se destacaram relatos dos sujeitos acerca de sua realidade, da possiblidade de romper estigmas, ressignificando conhecimento, ampliando percepções acerca dos temas exibidos e das possibilidades propiciadas pela tecnologia do cinema. Para compreender o fenômeno, as análises ancoram-se nos estudos sobre memória, direitos humanos, infância e cinema, por meio da pesquisa participante e da fenomenologia.Fil: Vieira e Silva, Ana Cristina.Fil: Gonçalves, Marlene

Síndrome de Wilkie : caso clínico

Trabalho Final do Curso de Mestrado Integrado em Medicina, Faculdade de Medicina, Universidade de Lisboa, 2016O síndrome de Wilkie ou síndrome da artéria mesentérica superior (SAMS) é uma situação clínica rara caracterizada pela obstrução parcial ou completa da terceira porção do duodeno pela artéria mesentérica superior na face anterior, e pela aorta e coluna vertebral posteriormente. Apresentamos um caso de clínico com dez anos de evolução, sendo diagnosticado SAMS através de meios complementares de diagnóstico. A terapêutica somente pôde ser cirúrgica devido à cronicidade. Após a cirurgia ficou assintomática e recuperou o peso. O síndrome de Wilkie é uma causa incomum de obstrução duodenal, com sintomas inespecíficos cujo diagnóstico requer um alto índice de suspeição pela equipa médica, pela história e sintomas clínicos, tendo maioria dos pacientes associado índice massa corporal baixo. As evidências imagiológicas fazem o diagnóstico definitivo.The Wilkie syndrome or superior mesenteric artery syndrome (SAMS) is a rare clinical situation characterized by partial or complete obstruction of the third portion of the duodenum by the superior mesenteric artery in the anterior face and by the aorta and spine on the posterior face. The following case presents a patient with ten years of evolution who was diagnosed with SAMS through complementary exams. Due to the chronicity of the disease, surgery was the only possible treatment. After surgery the patient became completely assymptomatic and recovered his weight. Wilkie syndrome is an uncommon cause of duodenal obstruction, with nonspecific symptoms, whose diagnosis depends on a high index of suspicion by the medical team, based on history and clinical symptoms, as the majority of cases present with a low body mass index. Imaging evidence make the definitive diagnosis

Development and Optimization of an Online SPE-HPLC-FD Method for Quantification of Fluoroquinolones in Wastewater Effluents

Fluoroquinolones are antimicrobial agents widely found in environmental matrices and extensively studied due to their persistence and implications for multiresistant bacteria. The presence of fluoroquinolones in the environment is mainly due to the incapability of wastewater treatment plants (WWTPs) to completely remove those compounds. The amount of fluoroquinolones released through effluents depends on the type of treatment used by the WWTPs. So, accurate analytical methods to quantify those compounds on WWTPs process and in effluents are crucial. Solid phase extraction (SPE) coupled to liquid chromatography is a straightforward technique that provides analyte extraction, cleanup, separation and detection while providing a good reproducibility and efficiency. The purpose of this work was the establishment of a novel method for quantification of Ofloxacin, Norfloxacin, Ciprofloxacin and Moxifloxacin on WWTPs effluents using on-line SPE. Samples were injected directly on a restricted access material column LichroCart 25-4 Lichrospher® RP-18 ADS (25 μm) and then transferred to an analytical column Luna PFP (2) (150 x 4.6 mm ID, 100 Å, 3 μm) for separation in isocratic mode with a mixture of 0.1% triethylamine in water (acidified to pH = 2.2 with trifluoroacetic acid) and ethanol as mobile phase; column oven was set at 45ºC. The detection was performed by fluorescence with an excitation wavelength of 290 nm and an emission wavelength of 460 nm. The injection volume of 100 μL of previous preconcentrated sample was compared with larger volume injection of only filtered effluent samples. The study was conducted with effluent samples collected from a municipal WWTP in the north of Portugal

Magnetic stimulation drives macrophage polarization in cell to–cell communication with IL-1β primed tendon cells

Inflammation is part of the natural healing response, but it has been simultaneously associated with tendon disorders, as persistent inflammatory events contribute to physiological changes that compromise tendon functions. The cellular interactions within a niche are extremely important for healing. While human tendon cells (hTDCs) are responsible for the maintenance of tendon matrix and turnover, macrophages regulate healing switching their functional phenotype to environmental stimuli. Thus, insights on the hTDCs and macrophages interactions can provide fundamental contributions on tendon repair mechanisms and on the inflammatory inputs in tendon disorders. We explored the crosstalk between macrophages and hTDCs using co-culture approaches in which hTDCs were previously stimulated with IL-1β. The potential modulatory effect of the pulsed electromagnetic field (PEMF) in macrophage-hTDCs communication was also investigated using the magnetic parameters identified in a previous work. The PEMF influences a macrophage pro-regenerative phenotype and favors the synthesis of anti-inflammatory mediators. These outcomes observed in cell contact co-cultures may be mediated by FAK signaling. The impact of the PEMF overcomes the effect of IL-1β-treated-hTDCs, supporting PEMF immunomodulatory actions on macrophages. This work highlights the relevance of intercellular communication in tendon healing and the beneficial role of the PEMF in guiding inflammatory responses toward regenerative strategies.This research was funded by the ERC CoG grant MagTendon (No. 772817), FCT Project Mag TT PTDC/CTM-CTM/29930/2017, and project NORTE-01-0145-FEDER-000021 supported by Norte Portugal Regional Operational Programme (NORTE 2020). A. Vinhas is funded by Fundação para a Ciência e Tecnologia (FCT)-doctoral grant PD/BD/128089/2016