Formation of Lipofuscin-Like Autofluorescent Granules in the Retinal Pigment Epithelium Requires Lysosome Dysfunction.

Purpose: We aim to characterize the pathways required for autofluorescent granule (AFG) formation by RPE cells using cultured monolayers. Methods: We fed RPE monolayers in culture with a single pulse of photoreceptor outer segments (POS). After 24 hours the cells started accumulating AFGs that were comparable to lipofuscin in vivo. Using this model, we used a variety of light and electron microscopical techniques, flow cytometry and Western blot to analyze the formation of AFGs. We also generated a mutant RPE line lacking cathepsin D by gene editing. Results: AFGs seem to derive from incompletely digested POS-containing phagosomes and after 3 days are surrounded by a single membrane positive for lysosome markers. We show by various methods that lysosome-phagosome fusion is required for AFG formation, and that impairment of lysosomal pH or catalytic activity, particularly cathepsin D activity, enhances AF accumulation. Conclusions: We conclude that lysosomal dysfunction results in incomplete POS degradation and enhanced AFG accumulation.

De seniores para seniores, Proalfa: um projeto de alfabetização com dinâmicas de animação / From seniors to seniors Proalfa: a literacy project with socio-educational animation dynamics

Num contexto de transformações demográficas, de incentivo à educação ao longo da vida, de necessidade de valorizar o papel dos seniores na sociedade e de promover a convivência entre gerações, foi criado o projeto ProAlfa, para utentes de uma estrutura residencial para pessoas idosas de Leiria ­ Portugal. O objetivo é potenciar a aprendizagem da leitura e da escrita, a partir de estratégias de animação socioeducativa, e promover a interação social. As oficinas decorrem uma vez por semana, no espaço da instituição, sendo dinamizadas, voluntariamente, por quatro estudantes seniores do Programa 60+ do Politécnico de Leiria. O presente artigo apresenta parte da investigação realizada no âmbito do ProAlfa, com os seguintes objetivos: conhecer os motivos que levam os seniores a participar no projeto, identificar as atividades que realizam e as que mais valorizam, além de conhecer as expetativas dos participantes sobre o mesmo. Em nível metodológico, a pesquisa é um estudo de caso, de abordagem mista concretizado a partir de um inquérito por entrevista, de cariz estruturado, aplicado a dezesseis participantes no ProAlfa. Estes são maioritariamente mulheres, têm idades compreendidas entre os 58 e os 102 anos, e são pouco escolarizados. Os resultados evidenciam que a oportunidade para a realização de aprendizagens valiosas e para conviver são os motivos apontados para a participação no projeto. Realizam semanalmente atividades de leitura, escrita e pintura, sendo esta última a mais valorizada. Apre- sentam expectativas reduzidas quanto à sua participação no projeto, nomeadamente em relação à aquisição de competências de leitura.(AU)

In a context of demographic changes, of encouraging lifelong education, of valuing the role of seniors in society and of promoting coexistence between generations, the ProAlfa project was created for users of a residential structure for elderly people in Leiria ­ Portugal. The objective is to enhance the learning of reading and writing, based on socio-educational animation strategies, and to promote social interaction. The workshops take place once a week, in the institution's space, being voluntarily promoted by four senior students of the Programa 60+ of the Polytechnic of Leiria. This article presents part of the research carried out within the scope of ProAlfa, with the following objectives: know the reasons that lead seniors to participate in the project, identify the activities they carry out and those that they value most, and to know the expectations of the participants about the project. At a methodological level, the research is a case study, with a mixed approach, obtained through an interview survey, of a structured nature, applied to 16 participants in ProAlfa. These are mostly women, ages between 58 and 102 years old, and with low level of education. The results show that the opportunity to learn valuable skills and to enjoy their time together are the reasons given for participating in the project. They carry out weekly reading, writing and painting sessions, the latter being the most valued. They have low expectations regarding their participation in the project, namely in relation to the acquisition of reading skills.(AU)

Intracranial Internal Carotid Artery Wall Calcification in Ischemic Strokes Treated with Thrombolysis.

BACKGROUND: Calcifications are an important element of atherosclerotic plaques and have been used as a marker of atherosclerosis and clinical outcome predictor in different vascular territories. CT-scan, performed in the acute ischemic stroke setting, can reliably detect intracranial arterial calcifications. OBJECTIVES: To investigate the association between intracranial internal carotid artery calcification and functional outcome, symptomatic intracerebral hemorrhage (sICH), recanalization, and death. METHODS: We included 396 consecutive ischemic stroke patients submitted to recombinant tissue plasminogen activator treatment between January 2011 and September 2014. Admission CT-scans were reviewed to calculate the Total Carotid Syphon Calcification score. Patients were followed for up to at least 6 months post-stroke or until death. Outcome measures included evaluation of recanalization on the first 24 h (transcranial color coded Doppler or angio-CT), sICH, and assessment of functional outcome at 3 months after stroke (using modified Rankin scale). RESULTS: Carotid artery wall calcification did not predict sICH, recanalization or any good outcome. However, it was a statistically significant predictor of death (OR 1.102, 95% CI [1.004-1.211], p = 0.042). DISCUSSION: Intracranial carotid artery calcification does not increase the risk of thrombolysis-induced sICH. Patients with higher grade of carotid artery wall calcification may have a higher mortality rate.