Neuronal deletion of GSK3beta increases microtubule speed in the growth cone and enhances axon regeneration via CRMP-2 and independently of MAP1B and CLASP2

BACKGROUND: In the adult central nervous system, axonal regeneration is abortive. Regulators of microtubule dynamics have emerged as attractive targets to promote axonal growth following injury as microtubule organization is pivotal for growth cone formation. In this study, we used conditioned neurons with high regenerative capacity to further dissect cytoskeletal mechanisms that might be involved in the gain of intrinsic axon growth capacity. RESULTS: Following a phospho-site broad signaling pathway screen, we found that in conditioned neurons with high regenerative capacity, decreased glycogen synthase kinase 3β (GSK3β) activity and increased microtubule growth speed in the growth cone were present. To investigate the importance of GSK3β regulation during axonal regeneration in vivo, we used three genetic mouse models with high, intermediate or no GSK3β activity in neurons. Following spinal cord injury, reduced GSK3β levels or complete neuronal deletion of GSK3β led to increased growth cone microtubule growth speed and promoted axon regeneration. While several microtubule-interacting proteins are GSK3β substrates, phospho-mimetic collapsin response mediator protein 2 (T/D-CRMP-2) was sufficient to decrease microtubule growth speed and neurite outgrowth of conditioned neurons and of GSK3β-depleted neurons, prevailing over the effect of decreased levels of phosphorylated microtubule-associated protein 1B (MAP1B) and through a mechanism unrelated to decreased levels of phosphorylated cytoplasmic linker associated protein 2 (CLASP2). In addition, phospho-resistant T/A-CRMP-2 counteracted the inhibitory myelin effect on neurite growth, further supporting the GSK3β-CRMP-2 relevance during axon regeneration. CONCLUSIONS: Our work shows that increased microtubule growth speed in the growth cone is present in conditions of increased axonal growth, and is achieved following inactivation of the GSK3β-CRMP-2 pathway, enhancing axon regeneration through the glial scar. In this context, our results support that a precise control of microtubule dynamics, specifically in the growth cone, is required to optimize axon regrowth

Avaliação, educação e comunicação do risco sísmico no contexto das escolas do Algarve

Tem sido observado, um pouco por todo o mundo, que as populações não têm consciência do risco sísmico dos edifícios onde habitam, e que possuem espectativas irrealistas sobre o comportamento dessas construções. A perceção generalizada é que as habitações irão ficar intactas após um sismo moderado ou destruidor, desde que estas tenham sido corretamente projetadas e construídas, de acordo com as normas vigentes. Contudo, os edifícios somente são projetados para garantir a salvaguarda da vida humana, e não para a salvaguarda do património em si, podendo até não ser economicamente viável a sua reparação. Além disso, um sismo moderado poderá causar danos moderados ou severos, principalmente a nível não-estrutural, afetando assim o funcionamento das instalações por tempo indeterminado. Por outro lado, é importante promover a educação e informação das populações para contribuir para o aumento da resiliência das sociedades a estes fenómenos. Neste contexto, foi desenvolvida uma plataforma informática no âmbito do projeto de investigação PERSISTAH (Projetos de Escolas Resilientes aos SISmos no Território do Algarve e de Huelva), que possibilita a realização de avaliações de segurança sísmica dos edifícios das escolas do primeiro ciclo do ensino básico existentes no Algarve, recorrendo a modernas técnicas de análise estrutural. Usando esta ferramenta, é possível verificar qual o grau de dano provável de um edifício de uma escola, quer seja para um qualquer cenário de ocorrência de um sismo, ou simplesmente no contexto da verificação dos níveis de segurança estabelecidos nas normas NP EN 1998-1:2010 e NP EN 1998-3:2017. Estão a ser criados documentos direcionados para técnicos da área da construção, para aprendizagem das referidas metodologias, assim como relativas ao uso de técnicas de reabilitação sísmica. Em paralelo, também foram desenvolvidos instrumentos para aumentar a resiliência sísmica da população estudantil, designadamente o Projeto Educativo “Porquê que o chão se move?”, com atividades e material pedagógico para a educação e comunicação do risco sísmico, e o “Guia Prático da Escola Resiliente aos Sismos”, que também visam a implementação de medidas de redução do risco sísmico não-estrutural.info:eu-repo/semantics/publishedVersio

Risk of low energy availability among female and male elite runners competing at the 26th European cross-country championships

Low energy availability (LEA) causes impaired physiological functioning. Cross-country running is a weight-sensitive sport, making athletes more prone to LEA. We aimed to estimate the prevalence of elite European cross-country athletes at risk of LEA using the LEA in Females Questionnaire (LEAF-Q) and to analyze demographic and physical characteristics that are associated with LEA. Athletes ≥ 18 years competing at the 26th European Cross-Country Championships (n = 602) were invited to complete a questionnaire (sociodemographic, training, anthropometric characteristics, and LEAF-Q). A total of 207 valid surveys were collected (83 females, 22.1 (4.0) years, and 124 males, 22.3 (4.1) years), and 16 surveys were excluded. A high prevalence of athletes at risk of LEA (64.3%) was observed, being higher in females than in males (79.5 and 54.0% respectively, p < 0.001). More than half of athletes (54.1%, n = 112) reported bowel movements once a week or more rarely, while 33 female athletes (41.3%) did not report normal menstruation. Overall, cross-country athletes are at high risk of LEA. Moreover, a high prevalence of gastrointestinal and menstrual impairments was reported. Hence, athletes should be followed by multidisciplinary teams to inform, prevent, and treat LEA and its effects.publishersversionpublishe

Promover a continuidade do ensino antes da ocorrência de um sismo

Portugal and Spain are countries prone to earthquake hazard. An earthquake that occurs in the sea may trigger a tsunami threat, so in this case they are also prone to tsunami. After a disaster, the size and severity of the event, undermines the availability of systems and services to function properly, resulting in losses and impacts in human, social, environmental and economic spheres. One of the potential negative effectsfor children’s education is caused by damaged school buildings and lack of road access to schools. The PERSISTAH project aims to study the seismic risk of primary schools located in the regions of Algarve (Portugal) and Huelva (Spain). The present research has specific objectives: 1) to improve school seismic safety; 2) to protect learners and education workers from death, injury and harm in schools; 3) to safeguard education investments; and 4) to strengthen risk reduction and resilience through educationThis project was funded by the INTERREG-POCTEP España-Portugal program and the European Regional Development Fund.info:eu-repo/semantics/publishedVersio

Familial Amyloid Polyneuropathy: Receptor for Advanced Glycation End Products-Dependent Triggering of Neuronal Inflammatory and Apoptotic Pathways

This is the publisher's version, also available electronically from http://www.jneurosci.org/content/21/19/7576.short?sid=590f7ac1-22ec-41bd-88e6-0995b772a4e0Familial amyloid polyneuropathy (FAP) is a neurodegenerative disorder associated with extracellular deposition of mutant transthyretin (TTR) amyloid fibrils, particularly in the peripheral nervous system. We have hypothesized that binding of TTR fibrils to the receptor for advanced glycation end products (RAGE) on critical cellular targets is associated with a destructive stress response underlying peripheral nerve dysfunction. Analysis of nerve biopsy samples from patients with FAP (n = 16) at different stages of disease (0–3), compared with age-matched controls (n = 4), by semiquantitative immunohistology andin situ hybridization showed increased levels of RAGE, beginning at the earliest stages of the disease (FAP 0;p < 0.02) and especially localized in axons. Upregulation of proinflammatory cytokines (tumor necrosis factor-α and interleukin-1β) (approximately threefold; p< 0.02) and the inducible form of nitric oxide synthase (iNOS) (∼2.5-fold; p < 0.04) was also observed in a distribution overlapping RAGE expression. Tyrosine nitration and increased activated caspase-3 in axons from FAP patients (p < 0.03) were apparent. Although these data suggest the presence of ongoing neuronal stress, there was no upregulation of neurotrophins (nerve growth factor and neurotrophin-3) in FAP nerves. Studies on cultured neuronal-like, Schwann, and endothelial cells incubated with TTR fibrils displayed RAGE-dependent expression of cytokines and iNOS at early times (6 and 12 hr, respectively), followed by later (24 hr) activation of caspase-3 and DNA fragmentation. We propose that the interaction of TTR fibrils with RAGE may contribute to cellular stress and toxicity in FAP. Furthermore, there is an apparent lack of responsiveness of Schwann cells in FAP nerve to provide neurotrophic factors

Multi-target analysis of cytostatics in hospital effluents over a 9-month period

The consumption of cytostatics, pharmaceuticals prescribed in chemotherapy, is increasing every year and worldwide, along with the incidence of cancer. The presence and the temporal evolution of cytostatics in wastewaters from a Portuguese hospital center was evaluated through a 9-month sampling campaign, comprising a total of one hundred and twenty-nine samples, collected from May 2019 to February 2020. Eleven cytostatics out of thirteen pharmaceuticals were studied, including flutamide, mycophenolate mofetil and mycophenolic acid, which have never been monitored before. Target analytes were extracted and quantified by solid-phase extraction coupled to liquid-chromatography-tandem mass spectrometry analysis; the method was fully validated. All pharmaceuticals were detected in at least one sample, bicalutamide being the one found with higher frequency (detected in all samples), followed by mycophenolic acid, which was also the compound detected at higher concentrations (up to 5340 ± 211 ng/L). Etoposide, classified as carcinogenic to humans, was detected in 60% of the samples at concentrations up to 142 ± 15 ng/L. The risk from exposure to cytostatics was estimated for aquatic organisms living in receiving bodies. Cyclophosphamide, doxorubicin, etoposide, flutamide, megestrol and mycophenolic acid are suspected to induce risk. Long-term and synergic effects should not be neglected, even for the cytostatics for which no risk was estimated.publishe

Sustained remission from depressive-like behavior depends on hippocampal neurogenesis

Impairment of hippocampal neurogenesis has been associated with the expression of depressive-like symptoms and some studies have suggested neurogenesis as a critical factor in the normalization of behavior by antidepressant (AD) drugs. This study provides robust evidence that ongoing neurogenesis is essential for the maintenance of behavioral homeostasis and that its pharmacological arrest precipitates symptoms commonly found in depressed patients. Further, the incorporation of newly born neurons and astrocytes into the preexisting hippocampal neurocircuitry is shown to be necessary for the spontaneous recovery from the adverse effects of stress and for long-term benefits of AD treatments.We thank M Carneiro and L Martins for technical assistance. AM-P, LP, MM and SM received fellowships from the Portuguese Foundation for Science and Technology (FCT). This work was supported by FCT (PTDC/SAU-NEU/105180/2008) and the ICVS

Avaliação, mitigação e comunicação do risco sísmico: ensinamentos do projeto PERSISTAH

Têm sido evidentes os efeitos dos sismos em países como Itália ou o México, onde ocorreram colapsos de edifícios escolares que provocaram vítimas mortais, os quais foram construídos, quer com estrutura de betão armado, quer em alvenaria tradicional. Algumas dessas escolas tinham sido objeto de intervenções de reabilitação, mas sem ter em conta a segurança sísmica. Em Portugal, também existem muitas escolas que foram sujeitas a intervenções visando melhorar as suas características térmicas, e tentando criar espaços mais modernos e funcionais, contudo sem existirem quaisquer preocupações em relação à segurança sísmica. Provavelmente, a recente legislação sobre reabilitação publicada em Portugal em 2019, que estabelece o Eurocódigo 8 (EC8) como sendo a base dos estudos de vulnerabilidade sísmica, irá lentamente começar a mudar as atitudes dos promotores imobiliários e dos técnicos ligados à construção civil, em relação ao risco sísmico. O projeto PERSISTAH, desenvolvido ao abrigo do programa INTERREG España-Portugal, foi pioneiro em Portugal na aplicação das metodologias de avaliação sísmica previstas no EC8, designadamente a um conjunto alargado de escolas do 1º ciclo do ensino básico existentes no Algarve. Para que fosse possível atingir esse ambicioso objetivo, foi desenvolvido um programa informático que permite avaliar o nível de segurança sísmica de uma escola para dois tipos de ação sísmica: a ação regulamentar estipulada no EC8 e a ação resultante de um determinado cenário de ocorrência de um sismo. Com este tipo de ferramenta informática, é possível hierarquizar as necessidades de reforço sísmico dos edifícios das escolas. A sua utilização permitiu verificar que as antigas escolas do Algarve, construídas em alvenaria de pedra, não satisfazem os níveis de segurança atualmente exigidos, o que é especialmente preocupante atendendo ao elevado número de crianças que ainda estudam nesse tipo de edifícios. Foi neste contexto que foi realizado o reforço sísmico de uma escola no Algarve, que se pretende que funcione como estudo-piloto, cujos ensinamentos serão incluídos num manual destinado a apoiar os profissionais que trabalham na região, em virtude deste novo desafio para o setor da construção civil do Algarve. No entanto, também é importante que as populações entendam que mesmo que um edifício cumpra os requisitos de segurança sísmica estipulados num regulamento, as estruturas não são concebidas para resistir aos sismos sem danos, podendo até mesmo ser economicamente inviável a sua reparação após um sismo muito intenso. Uma estrutura bem projetada somente tem como objetivo garantir a salvaguarda da vida humana. Neste contexto, a comunicação do risco sísmico é de grande importância para o aumento da resiliência sísmica das comunidades, sendo que a população estudantil constitui um alvo muito importante. Por esse motivo, foram criados textos de apoio aos professores e alunos, de modo a facilitar a assimilação de conceitos e de medidas de autoproteção, designadamente relacionadas com os elementos não-estruturais, que poderão ficar seriamente danificados mesmo em edifícios corretamente projetados, podendo causar ferimentos ou mesmo a perda de vidas e funções de um edifício. O projeto PERSISTAH permitiu perceber quais as reais dificuldades associadas à avaliação, mitigação e comunicação do risco sísmico, cujos ensinamentos são descritos no presente trabalho.Programa INTERREG-POCTEP España-Portugal e o Fundo Europeu de Desenvolvimento Regional (FEDER); Projeto PERSISTAH (Projetos de Escolas Resilientes aos Sismos no Território do Algarve e de Huelva)info:eu-repo/semantics/publishedVersio

Transboundary Impact Assessment in the north-western Iberian Peninsula

En el marco del proyecto SIMAtlantic, se ha llevado a cabo un caso de estudio entre España y Portugal. Este documento describe una metodología propuesta para la Evaluación de Efectos Acumulativos con el fin de evaluar los impactos/efectos acumulativos a escala transfronteriza. El trabajo se llevó a cabo en un caso de estudio específico en la región noroeste de la Península Ibérica, en una zona transfronteriza entre Portugal y España

Rotator cuff isokinetic strength of young group aged swimmers in a competitive training program

Introduction: In any given sports activity, muscular or movement instability is pointed out as having a high influence for the maintenance of correct joint functionality. In swimming, due to the repetitive use of shoulder muscles, there is a higher tendency to promote injuries at the shoulder joint, especially when instability between the internal (IR) and external (ER) shoulder rotators is present. The aims of this study were to evaluate the rotator cuff isokinetic strength in order to determine any possible muscular over compensation or imbalance, and to assess differences between the dominant and non-dominant side, of young group aged swimmers. Methods: Forty young aged swimmers in a competitive training program were evaluated, and divided in two groups. Only subjects without any previous history of strength training were included. Group 1 (n=19), age 15 to 16 (years); height [mean (SD)] 168.6 (6.5) (centimeters); weight 59.6 (6.0) (kilograms), training/week 113.7 (12.6) (minutes); experience 5.3 (1.6) (years) and Group 2 (n=21); age 13 to 14; height 160.5 (12.2); weight 50.4 (10.8); training/week 88.6 (17.7); experience 4.1 (2.2) (years). The maximal unilateral isokinetic strength was measured (Biodex System 3) on the shoulder IR and ER during concentric action at 60°/second, bilaterally. Data was examined by the application of appropriate statistical tests for the analysis of variance. Results: Statistical differences were found on peak torque (Nm) at the agonist/antagonist ratio measured unilaterally [Group 1: right shoulder 35.8 (7.9) (IR) and 25.8 (4.7) (ER) (p=0.001) and left shoulder 32.7 (6.7) (IR) and 23.9 (5.1) (ER) (p=0.001) / Group 2: right shoulder 23.4 (6.5) (IR) and 19.0 (5.4) (ER) (p=0.048) and left shoulder 22.4 (5.4) (IR) and 16.8 (6.1) (ER) (p=0.027)]. However, no statistical differences were found between the maximal rotation strength measured bilaterally [Group 1: ER (p=0.280) and IR (p=0.246) / Group 2: ER (p=0.186) and IR (p=0.522)]. Conclusion: Young swimmers involved in a competitive training program can suffer from unilateral over compensation at shoulder IR or ER. This compensation seems to increase with the number of training years. To enable a higher stability between the internal and external shoulder rotators it is recommended that shoulder muscle strengthening should be considered in young aged swimmers training programs