Exploring the astrocytic neuroprotective functions in a chronic mild stredd model of depression

Dissertação de mestrado em Ciências da SaúdeDepression is a multidimensional psychiatric disorder that affects millions of people worldwide. In order to unveil the pathophysiology of this disorder, increasing significance is being given to the study of the abnormal neurotransmission in brain areas affected in depression. Particularly, glutamatergic excitotoxicity has been suggested as one of the possible process underlying the installation of the disease. Previous studies have suggested that glutamate release is increased in depressed subjects. Additionally, the β-lactamic antibiotic ceftriaxone (CEF) was described to increase glutamate uptake by the glutamate transporter GLT-1 (expressed mainly in astrocytes), preventing cellular damage. Taking these findings into account, the main goal of this work was to understand whether CEF-triggered enhancement of glutamate uptake might be used to prevent or reverse the deleterious effects of glutamatergic excitotoxicity in an animal model of depression, the unpredictable chronic mild stress (uCMS). For this, animals were subdivided in two different main sets - Prevention and Treatment. Different groups of animals were administered either with CEF and/or two antidepressants (ADs; fluoxetine and imipramine). CEF administration, at an early stage, prevented the installation of depressive-like behavior, yet it failed to reverse the installed depressive phenotype induced by uCMS exposure. CEF seems to reverse partially the cognitive deficits caused by uCMS exposure. Analysis of the collected brain tissue revealed that the prevention of the depressive-like behavior was correlated with an increased GLT-1 gene expression in the ventral hippocampus and with an increased expression of GLT-1 transporter in the dorsal dentate gyrus (DG) of the hippocampus. Morphological analyses disclosed neuronal atrophy of the DG granule neurons in the dorsal DG after uCMS exposure, which was prevented by CEF administration. These results suggest that CEF administration promotes GLT-1 transporter up-regulation, which may prevent excitotoxicity processes at glutamatergic synapses in the hippocampus, thus preventing the installation of the depressive-like behaviors triggered by uCMS. These observations elucidate the potential use of CEF, or similar drugs, in the prevention of depressive behavior, paving the way for the development of new therapeutic strategies.A depressão é uma doença psiquiátrica que afeta milhões de pessoas em todo o mundo. Na tentativa de compreender os mecanismos fisiopatológicos da doença, a comunidade científica tem-se dedicado ao estudo de defeitos na neurotransmissão em áreas cerebrais afetadas na doença. Em particular, a excitotoxicidade glutamatérgica tem sido sugerida como um processo subjacente à instalação da doença. Estudos anteriores verificaram que a libertação de glutamato está aumentada em indivíduos deprimidos. Neste contexto, foi também descrito que o antibiótico β-lactâmico ceftriaxona (CEF) aumenta a captação de glutamato pelo transportador GLT-1 (expresso principalmente em astrócitos), prevenindo os danos celulares. Considerando estas observações, o objetivo deste trabalho foi compreender se o aumento da captação de glutamato desencadeado pela CEF poderá prevenir ou reverter os efeitos deletérios da excitotoxicidade glutamatérgica num modelo animal de depressão, de exposição crónica ao stress (uCMS). De acordo com o objectivo, os animais foram divididos em dois grupos principais – Prevenção e Tratamento. A diferentes subgrupos de animais foram administrados CEF e/ou dois antidepressivos (fluoxetina e imipramina). A administração de CEF preveniu o desenvolvimento do comportamento depressivo, no entanto foi ineficaz na reversão deste fenótipo já instalado após exposição crónica a stress. Adicionalmente, a CEF parece reverter parcialmente os défices de cognição induzidos pelo uCMS. Análises do tecido cerebral dos animais revelaram uma correlação entre a prevenção do comportamento depressivo e a expressão do gene GLT-1 no hipocampo ventral e do transportador GLT-1 no girus denteado (GD) dorsal. A avaliação da morfologia dos neurónios granulares do GD revelou que a CEF foi capaz de prevenir a atrofia dendrítica no GD dorsal provocada pela exposição ao stress. Estes resultados sugerem que a administração de CEF estimula a expressão de GLT-1, prevenindo os eventos excitotóxicos nas sinapses glutamatérgicas no hipocampo e o consequente desenvolvimento do comportamento depressivo causado pela exposição ao stress crónico. Estas observações indicam um potencial efeito da CEF ou de outros fármacos que promovam a expressão do GLT-1, na prevenção do desenvolvimento de comportamento depressivo, abrindo caminho para o desenvolvimento de novas possibilidades terapêuticas

Análise comparativa de soluções de reabilitação nas margens do Rio Tinto (Gondomar)

Trabalho realizado na Câmara Municipal de Gondomar, orient. pela Drª Iva Carla Vieira Rodrigues FerreiraTese de mestrado integrado. Engenharia do Ambiente. Faculdade de Engenharia. Universidade do Porto. 201

Molecular techniques and target selection for the identification of Candida spp. in oral samples

Candida species are the causative agent of oral candidiasis, with medical devices being platforms for yeast anchoring and tissue colonization. Identifying the infectious agent involved in candidiasis avoids an empirical prescription of antifungal drugs. The application of high-throughput technologies to the diagnosis of yeast pathogens has clear advantages in sensitivity, accuracy, and speed. Yet, conventional techniques for the identification of Candida isolates are still routine in clinical and research settings. Molecular approaches are the focus of intensive research, but conversion into clinic settings requires overcoming important challenges. Several molecular approaches can accurately identify Candida spp.: Polymerase Chain Reaction, Microarray, High-Resolution Melting Analysis, Multi-Locus Sequence Typing, Restriction Fragment Length Polymorphism, Loop-mediated Isothermal Amplification, Matrix Assisted Laser Desorption Ionization-mass spectrometry, and Next Generation Sequencing. This review examines the advantages and disadvantages of the current molecular methods used for Candida spp. Identification, with a special focus on oral candidiasis. Discussion regarding their application for the diagnosis of oral infections aims to identify the most rapid, affordable, accurate, and easy-to-perform molecular techniques to be used as a point-of-care testing method. Special emphasis is given to the difficulties that health care professionals need to overcome to provide an accurate diagnosis.info:eu-repo/semantics/publishedVersio

Hyperglycaemic Hemichorea

Background: Hyperkinetic disorders such as hemichorea can be caused by cerebrovascular, infectious or inflammatory diseases or by metabolic conditions such as hyperglycaemia. Hyperglycaemic hemichorea is a rare movement disorder which is frequently misdiagnosed. It is characterized by involuntary, continuous, non-patterned movements on one side of the body, basal ganglia lesions seen on head CT or MRI, and clinical improvement after blood glucose normalization. We describe the case of a female patient with uncontrolled diabetes who presented with hemichorea. Case presentation: We report the case of a 69-year-old woman with type 2 diabetes who presented with abnormal movements of the right upper limb. She had no neurological signs other than hemichorea. Her blood glucose level was 349 mg/dl and her glycosylated haemoglobin level (HbA1c) was 10.5%. Head CT and MRI showed no changes in the basal ganglia or ischaemic lesions. The patient was started on insulin and haloperidol with clinical improvement. Conclusion: Larger case series are needed to establish better understanding of the physiopathological mechanisms and diagnostic criteria of hyperglycaemic hemichorea. The most important diagnostic criterion is clinical improvement after glycaemic control

Estudo Nacional sobre a Violência no Namoro em Contexto Universitário: Crenças e Práticas – 2017/2020

info:eu-repo/semantics/publishedVersio

Efeito de um programa de tiro com zarabatana na função respiratória de adultos com Dificuldades Intelectuais e Desenvolvimentais

Introdução e Objetivos: A principal causa de morte e de internamento em pessoas com Dificuldades Intelectuais e Desenvolvimentais (DID) é a patologia respiratória. O treino de tiro com zarabatana tem o potencial de melhorar a função respiratória de populações com/sem patologia através de uma atividade lúdica com significado, que envolve todo o ciclo respiratório. No entanto, o seu impacto na população com DID não é claro. Este estudo explorou os efeitos de um programa de tiro com zarabatana na função respiratória de adultos com DID. Material e Métodos: Foram recrutados 16 adultos com DID na Organização de Apoio e Solidariedade para a Integração Social (OASIS) e distribuídos, de acordo com a disponibilidade para participar nas sessões, em 2 grupos: intervenção (tiro com zarabatana, GI, n=8) e controlo (usual care, GC, n=8). O programa de tiro com zarabatana foi realizado na OASIS 1 vez/semana durante 3 meses (início: 30-40 tiros a 4m do alvo; progressão: aumento da distância/n.º tiros). Foram avaliadas: 1) a função pulmonar, através do Volume Expiratório Forçado no 1º segundo (FEV1%previsto), Capacidade Vital Forçada (FVC%previsto) e Pico de Fluxo Expiratório (PEF); 2) a força dos músculos respiratórios através das Pressões Inspiratória (PIM) e Expiratória (PEM) Máximas. Foram analisadas as diferenças: 1) entre GI e GC na baseline (M0) e aos 3 meses (M1) (teste t-student ou teste Mann-Whitney; α=0,05) e 2) entre os momentos de avaliação (M0-M1) em cada grupo (teste Wilcoxon; α=0,05). Resultados: 12 participantes concluíram o estudo, 7 no GI (33,0±14,4 anos; 5 mulheres) e 5 no GC (51,8±9,3 anos; 3 mulheres). Os grupos apresentaram uma diferença estatisticamente significativa em relação à idade (t-student, p=0.029), mas não em relação ao sexo. Os grupos não apresentaram diferenças significativas em M0 (p>0.05). Não foram observadas diferenças significativas na função pulmonar e na força dos músculos respiratórios (p>0,05) em M1 entre o GI e o GC (FEV1-GI=62,3±14,6%previsto FEV1-GC=79,0±17,7%previsto; FVCGI=64,1±17,2%previsto FVCGC=70,8±19,4%previsto; PEFGI=146,3±36,0L/min PEFGC= 279,0±166,9L/min; PIMGI=28,0±22,3cmH2O PIMGC=33,6±13,1cmH2O; PEMGI= 38,4±25,9cmH2O PEMGC= 40,4±16,6cmH2O). Nenhum dos grupos apresentou diferenças entre M0 e M1 (p<0,05). Conclusões: O treino de tiro com zarabatana não parece produzir efeitos significativos na função respiratória em adultos com DID a curto prazo. No entanto, são necessários mais estudos com desenhos robustos para confirmar os resultados.N/

Teria a Cistatina C um papel como substituto metabólico na diálise peritoneal além de sua associação com a função renal residual?

Introduction: It has been suggested that cystatin C levels are modified by obesity and inflammation. Furthermore, cystatin C has been associated with cardiovascular events and mortality outcomes. Aim: To study the association of cystatin C with the metabolic profile and cardiovascular disease of peritoneal dialysis patients. Methods: Data collected included clinical, laboratorial, and multifrequency bioimpedance assessment of 52 stable peritoneal dialysis patients. Minimal residual renal function was defined as > 2mL/min/1.73m2. Results: Serum cystatin C was not significantly associated with peritoneal or urinary cystatin C excretion. Negative correlation of cystatin C with normalized protein catabolic rate (rho -0.33, p = 0.02) and a trend towards positive correlation with relative body fat (rho 0.27, p = 0.05) were not independent from residual renal function. Cystatin C was not significantly associated with cardiovascular disease (p = 0.28), nor with glycated hemoglobin (p = 0.19) or c-reactive protein (p = 0.56). In the multivariate model, both age and diabetes were the strongest predictors of cardiovascular disease (odds ratio 1.09, p = 0.029 and odds ratio 29.95, p = 0.016, respectively), while relative body fat was negatively associated with cardiovascular disease (p = 0.038); neither cystatin C (p = 0.096) nor minimal residual renal function (p = 0.756) reached a significant association with cardiovascular disease. Conclusions: In this group of peritoneal dialysis patients, cystatin C did not correlate with the metabolic or inflammatory status, nor cardiovascular disease, after adjustment for residual renal function.Introdução:: Tem sido sugerido que os níveis de cistatina C são modificados pela obesidade e inflamação. Além disso, a cistatina C tem sido associada a eventos cardiovasculares e desfechos de mortalidade. Objetivo:: Estudar a associação da cistatina C com o perfil metabólico e doença cardiovascular de pacientes em diálise peritoneal. Métodos:: Os dados coletados incluíram avaliação clínica, laboratorial e de bioimpedância múltipla de 52 pacientes estáveis em diálise peritoneal. A função renal residual mínima foi definida como > 2mL/min/1,73m2. Resultados:: A cistatina C sérica não esteve significativamente associada à excreção peritoneal ou urinária. A correlação negativa da cistatina C com a taxa catabólica protéica normalizada (rho -0,33, p = 0,02) e uma tendência de correlação positiva com a gordura corporal relativa (rho 0,27, p = 0,05) não foram independentes da função renal residual. A cistatina C não se associou significativamente à doença cardiovascular (p = 0,28), nem com hemoglobina glicada (p = 0,19) ou proteína C reativa (p = 0,56). No modelo multivariado, idade e diabetes foram os mais fortes preditores de doença cardiovascular (razões de probabilidade 1,09, p = 0,029 e 29,95, p = 0,016, respectivamente) enquanto a gordura corporal relativa se associou negativamente à doença cardiovascular (p = 0,038). A cistatina C não se associou significativamente com doença cardiovascular (p = 0,096), tampouco a função residual mínima (p = 0,756). Conclusão:: Neste grupo de pacientes em diálise peritoneal, a cistatina C não se correlacionou com o estado metabólico ou inflamatório, nem com doença cardiovascular, após ajuste para função renal residual.info:eu-repo/semantics/publishedVersio

Population wide testing pooling strategy for SARS-CoV-2 detection using saliva

SARS-CoV-2 pandemic has forced frequent testing of populations. It is necessary to identify the most cost-effective strategies for the detection of COVID-19 outbreaks. Nasopharyngeal samples have been used for SARS-CoV-2 detection but require a healthcare professional to collect the sample and cause discomfort and pain to the individual. Saliva has been suggested as an appropriate fluid for the diagnosis of COVID-19. We have investigated the possibility of using pools of saliva samples to detect SARS-CoV-2 in symptomatic and asymptomatic patients. Two hundred and seventy-nine saliva samples were analyzed through RT-PCR of Envelope, Nucleocapsid and Open Reading Frame 1ab genes. Reproducibility assays showed an almost perfect agreement as well as high sensitivity (96.6%), specificity (96.8%), positive predicted value (96.6%), and negative predicted value (96.8%). The average Cycle Threshold of the genes detected was 29.7. No significant differences (p > 0.05) were detected when comparing the cycle threshold average of two consecutive reactions on the same positive saliva samples. Saliva samples have a higher median viral load (32.6) than in nasopharyngeal samples (28.9), although no significant differences were detected (p > 0.05). Saliva-pool samples allowed effective SARS-CoV-2 screening, with a higher sensibility (96.9%) on 10-sample pools than in 20-sample pools (87.5%). Regardless of pools size specificity was high (99.9%) and an almost perfect agreement was observed. Our strategy was successfully applied in population wide testing of more than 2000 individuals, showing that it is possible to use pooled saliva as diagnostic fluid for SARS-CoV-2 infection.info:eu-repo/semantics/publishedVersio

Preclinical assessment of mesenchymal-stem-cell-based therapies in spinocerebellar ataxia type 3

The low regeneration potential of the central nervous system (CNS) represents a challenge for the development of new therapeutic strategies for neurodegenerative diseases, including spinocerebellar ataxias. Spinocerebellar ataxia type 3 (SCA3)—or Machado–Joseph disease (MJD)—is the most common dominant ataxia, being mainly characterized by motor deficits; however, SCA3/MJD has a complex and heterogeneous pathophysiology, involving many CNS brain regions, contributing to the lack of effective therapies. Mesenchymal stem cells (MSCs) have been proposed as a potential therapeutic tool for CNS disorders. Beyond their differentiation potential, MSCs secrete a broad range of neuroregulatory factors that can promote relevant neuroprotective and immunomodulatory actions in different pathophysiological contexts. The objective of this work was to study the effects of (1) human MSC transplantation and (2) human MSC secretome (CM) administration on disease progression in vivo, using the CMVMJD135 mouse model of SCA3/MJD. Our results showed that a single CM administration was more beneficial than MSC transplantation—particularly in the cerebellum and basal ganglia—while no motor improvement was observed when these cell-based therapeutic approaches were applied in the spinal cord. However, the effects observed were mild and transient, suggesting that continuous or repeated administration would be needed, which should be further tested.This research was funded by the National Ataxia Foundation (NAF) and by Portuguese national funds, through the Foundation for Science and Technology (FCT)—projects UIDB/50026/2020, UIDP/50026/2020, POCI-01-0145-FEDER-029206, and through the Santa Casa Neuroscience Awards (Santa Casa da Misericórdia Lisboa)—project MC-04/17. Additionally, this project was funded by the ICVS Scientific Microscopy Platform, a member of the national infrastructure PPBI—Portuguese Platform of Bioimaging (PPBI-POCI-01-0145-FEDER-022122). S.C.S. received an individual fellowship within the project TUBITAK/0007/2014. The FCT funded individual fellowships to J.S C., A.N.-C., B.M.- P., F.G.T., R.L., S.M., N.A.S., C.S.-C., and S.D.-S. (SFRH/BD/140624/2018, SFRH/BPD/118779/2016, SFRH/BD/120124/2016, SFRH/BPD/118408/2016, PD/BDE/127836/2016, CEECIND/01902/2017, CEECIND/04794/2017, CEECIND/03887/2017, and CEECIND/00685/2020)