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- Deciphering neuroimmune interactions in alcohol intake in mouse model of intermittent access in male and femalePublication . Pacheco, Raquel; Canedo, Teresa; Rodrigues, Ana M.; Moreira, Joana; Relvas, João B.; Socodato, Renato; Summavielle, Teresa; Summavielle, TeresaExcessive alcohol consumption continues to pose a significant global health challenge, with detrimental effects on millions of individuals. Our laboratory has shown that alcohol exposure triggers reactive changes in astrocytes, including alterations in gene expression, activity, and proliferation, while also affecting microglial morphology and immune responses. We are to characterizing the effects of chronic alcohol consumption using a wellestablished voluntary alcohol drinking model in adult mice, to investigate the impact of chronic alcohol exposure on the prefrontal cortex (PFC), focusing on glial cell morphology, synaptic density, and behaviour. Mice are exposed to intermittent “every-other-day” (EOD) access to alcohol 15% (v/v) for 3 weeks, and behaviourally tested for anxiety, depression and memory, before sacrifice at 21 days of alcohol, or at 7 days of withdrawal. Brains were processed for glial cell analysis. Our preliminary findings revealed sex-specific responses following chronic alcohol exposure. Male mice exhibited increased astrocyte volume in the ventromedial PFC (vmPFC) and hyper-ramification in the ventrolateral PFC (vlPFC), whereas females displayed reductions in astrocyte size and complexity. Microglia morphology also differed between sexes, with females showing increased cell volume and males displaying reduced microglial volume in the vlPFC. These results suggest distinctive immune and synaptic responses to ethanol in males and females. Of note, we observed heightened inhibitory synapse density in the male PFC, while females exhibited increased excitatory synapse density. We are now conducting a proteomic analysis of PFC synaptosomes to identify important molecular targets in the crosstalk between neuros and glial cells. With this work we expect to clarify the complex interplay between chronic ethanol exposure, sex, and PFC function, find also new targets for innovative therapeutic approaches.
- Early-life exposure to non-nutritive sweeteners: effects on hepatic FGF21 pathways and mitochondrial functionPublication . Brandão, Sofia; Bracchi, Isabella; Felipe, Stela Mirla; Magalhães, José; Keating, Elisa; Negrão, RitaTo combat obesity, diets and food products containing non-nutritive sweeteners (NNS), which provide sweetness without caloric intake, have gained popularity, even among pregnant women who reported intentional NNS consumption during gestation. In 2023, the World Health Organization warned about NNS consumption, as their long-term effects may be associated with an increased risk of type 2 diabetes, cardiovascular diseases, and mortality. Importantly, the environment during early life can induce permanent changes in metabolic health in adult life, suggesting that fetal exposure to NNS may induce long-term metabolic dysfunction in offspring. Fibroblast growth factor 21 (FGF21) is a key regulator of metabolic homeostasis. So, this branch of the MHSWEET project aims to study the effects of Rebaudioside A (RebA, the major sweetener component of stevia) exposure during the perigestational period on hepatic lipid metabolism of adult offspring and the involvement of FGF21. Female Sprague-Dawley rats were administered RebA in the drinking water or water as control from 4 weeks before mating until weaning. Food and beverage consumption, morphometric parameters, fasting glucose, oral glucose tolerance, and insulin sensitivity were measured before sacrifice at 10 months of age. Functionality of G1 hepatic mitochondria (OROBOROS® Oxygraph-2k system), hepatic triglyceride levels (Oil-Red-O staining), genes involved in hepatic lipid metabolic pathways and inflammation (RT-PCR) will be evaluated. The results obtained so far, showed that RebA exposure increased body weight in adult female offspring and induced dysglycemia in adult male offspring. The remaining studies are still being conducted. Results seem to support the hypothesis that intrauterine exposure to RebA may program offspring to a dysmetabolic state. These results will be useful to improve knowledge, and support public health policies regarding NNS use throughout life.
- Proteomics characterization of human uterine samples during reproductive ageingPublication . Dias, Bruna; Formoso, Inês; Guedes-Martins, Luís; Almeida, Henrique; Silva, Elisabete; Silva, André M.N.; Matos, LilianaNowadays, postponing motherhood has become more common in modern societies. Such decision rises major social and health concerns, as advanced maternal age associates with low fertility, pregnancy complications and a greater need for assisted reproductive techniques. Female reproductive ageing is associated with alterations that are particularly evident in the ovaries. Yet, given the pivotal contribution of the uterus to embryo adhesion, implantation, and fetal development, we believe that age-dependent uterine changes impair fertility. Evidence in mice already reported age-associated uterine changes. Also, albumin carbonylation was found increased in uterine samples from older term-pregnant women. Moreover, preliminary data show changes in composition and oxidation status of uterine extracellular matrix proteins with higher maternal age. We hypothesize that the uterine proteome changes during reproductive ageing, impairing tissue structure and function. This study aims to identify age-related alterations in uterine protein composition through a complete proteomics approach. Uterine samples were collected during c-section from termpregnant women and immediately fixed for paraffin-embedding or frozen for molecular analysis. Six samples were selected: three from younger and three from reproductively aged women. Paraffin-embedded samples were histologically analyzed after H+E staining. Protein lysates were obtained after sample homogenization and total protein content was determined and were then separated by SDS-PAGE, stained with Coomassie Blue, and will be subjected to a MS based proteomics study. Histological analysis of the samples has proven their uterine origin. SDS-PAGE results confirmed the successful protein extraction. The identification of uterine proteins differentially expressed according to maternal age will allow the development of therapeutic approaches, aiming to attenuate age-associated female fertility decline.
- Obesity and physical loading during manual liftingPublication . Colim, Ana; Arezes, Pedro; Flores, Paulo; Monteiro, Pedro; Mesquita, InêsManual lifting greatly increases the risk of developing work-related musculoskeletal disorders (WRMSD). Muscle loading related to manual tasks may be significantly influenced by workers’ body conditions, being obesity one of them. In the present study, different tasks of manual lifting were performed by 14 participants with different obesity levels, defined in terms of individuals’ fat mass percentage. Surface electromyography (EMG) data were collected bilaterally from 3 muscles recruited during this type of tasks. EMG data normalization was based on the percentage of maximum contraction during each task (MCT). Furthermore, the participants reported physical loading by using the Rated Perceived Exertion scale (RPE). These techniques sought to analyze the physical loading during manual lifting tasks. The obtained results indicate that obesity level influenced the MCT percentage, which in turn increases the muscle loading during manual lifting tasks. However, obesity level does not seem to influence the RPE values. In this context, further studies based on biomechanical approaches are required to provide a more complete understanding of the obesity effects on musculoskeletal loading during manual lifting.
- Muscle loading and psychophysical effects of obesity during vertical handling tasksPublication . Colim, Ana; Arezes, Pedro; Flores, Paulo; Monteiro, Pedro; Mesquita, InêsIt has been recognized by many researchers that the prevalence of obesity is a serious public health problem. Several physical problems are associated with obesity, which can negatively affect the individuals’ work performance. In occupational contexts, vertical handling tasks, including lifting and/or lowering loads, are common and produce significant muscle loading. For these reasons, Surface electromyography (EMG) data were collected bilaterally from muscles recruited during these tasks. Additionally, the participants reported physical loading by using the Rated Perceived Exertion scale (RPE). These techniques sought to analyze the physical loading during vertical handling tasks. The obtained results indicate that obesity seem to increase the muscle loading during vertical handling tasks. However, obesity level does not seem to influence the RPE values.
- Efeito de uma sessão de exercício físico associao à transferência elétrica capacitiva resistivaPublication . Noites, Andreia; Vale, Ana Luísa Pinto; Pereira, Ana Sofia Teixeira; Morais, Andreia Sofia Silva; Amorim, Manuela; Moreira, Teresa; Vilarinho, RuiA acumulação de tecido adiposo na região abdominal é um fator de risco para o desenvolvimento de doenças cardiovasculares. No sentido de procurar estratégias que atuem sobre a adiposidade abdominal, a radiofrequência e o exercício físico surgem como aliados, permitindo elevar o metabolismo celular potenciando a lipólise. Verificar se uma sessão de Transferência Elétrica Capacitiva abdominal associada a uma sessão de exercício físico aeróbio moderado é mais eficaz ao nível da lipólise do que uma sessão de exercício físico isolado. Este estudo randomizado controlado duplamente cego, contou com a participação de 30 indivíduos saudáveis do sexo feminino, distribuídos aleatoriamente em dois grupos, um grupo experimental (n=15) e um grupo placebo (n= 15). Os participantes alocados ao grupo experimental foram sujeitos a uma sessão de Transferência Elétrica Capacitiva abdominal seguida de exercício físico aeróbio de intensidade moderada. O grupo placebo realizou o protocolo de RF com intensidade de zero watts, seguido do protocolo de exercício físico aeróbio semelhante ao do grupo experimental. Foram realizadas análises sanguíneas antes e após o protocolo de intervenção de forma a determinar alterações no perfil lipídico e na concentração de glicerol. O estudo foi aprovado pela comissão de ética da Escola Superior de Saúde- Politécnico do Porto (registo nº 1351/2018). Não foram observadas alterações significativas nas concentrações de glicerol entre os grupos (p>0,05), verificando-se, entre momentos, um aumento estatisticamente significativo desta variável nos dois grupos. Não foram detectadas alterações nas restantes componentes lipídicas entre grupos após intervenção e entre os dois momentos de avaliação (p>0,05). Não se verificou o aumento da lipólise quando se associou a Transferência Elétrica Capacitiva Resistiva ao exercício físico, sugerindo que o efeito desta técnica pode ocorrer por outros mecanismos.
- Efeitos do exercício físico aeróbio combinado com tranferência elétrica capacitiva E R.Publication . Noites, Andreia; Vale, Ana Luísa Pinto; Pereira, Ana Sofia Teixeira; Morais, Andreia Sofia Silva; Carvalho, Paulo; Mendonça, Adriana; Vilarinho, RuiA acumulação de tecido adiposo na região abdominal é um fator de risco para o desenvolvimento de doenças cardiovasculares. No sentido de procurar estratégias que atuem sobre a adiposidade abdominal, a Transferência Elétrica Capacitiva Resistiva e o exercício físico surgem como aliados, permitindo intervir nas medidas antropométricas, indicadoras desse risco. Verificar as diferenças provocadas nas medidas antropométricas por quatro sessões de um protocolo de transferência elétrica capacitiva resistiva abdominal combinada com exercício físico aeróbio, quando comparado com exercício físico aeróbio isolado. ambos os protocolos após quatro sessões nas medidas antropométricas. Este estudo randomizado controlado duplamente cego, contou com a participação de 28 indivíduos, distribuídos aleatoriamente em dois grupos, o grupo experimental (n=14) e placebo (n=14). Foram realizadas 4 sessões, com um intervalo mínimo de 24 horas entre elas. Os participantes alocados ao grupo experimental foram sujeitos a uma sessão de Transferência Elétrica Capacitiva abdominal seguida de exercício físico aeróbio de intensidade moderada. O grupo placebo realizou o protocolo de Transferência Elétrica Capacitiva com intensidade de zero watts, seguido do protocolo de exercício físico aeróbio semelhante ao do grupo experimental. Procedeu-se à avaliação das medidas antropométricas, nomeadamente perímetro da cintura, rácio cintura/altura e à medição da espessura de tecido adiposo subcutâneo com recurso à ultrassonografia, antes do início da intervenção e imediatamente após a quarta sessão. O estudo foi aprovado pela comissão de ética da Escola Superior de Saúde- Politécnico do Porto (registo nrº 1351/2018). Observaram-se diferenças estatisticamente significativas em todas as medidas (p<0,05), sendo que o GE apresentou uma diminuição significativa destas variáveis (p<0,05). A associação da radiofrequência com o exercício físico parece ser um meio coadjuvante na redução do tecido adiposo abdominal em indivíduos do sexo feminino saudáveis.
- Effects of embryonic exposure to venlafaxine on a zebrafish modelPublication . Barros, Eduarda; Abreu, Isabel; Guimarães, Laura; Melo, PedroMajor depressive disorder affects over 350 million people around the world, and might result from a complex interaction of epigenetic, genetic, environmental and developmental factors. Antidepressants are a class of neuroactive compounds that are used mostly in the treatment of clinically severe mood and anxiety disorders, and can be divided into three major classes: tricyclic antidepressants, monoamine oxidase inhibitors and selective serotonin reuptake inhibitors. The boost on antidepressants’ prescription and consumption is related to an increase in the prevalence of psychiatric disorders and knowledge of mental health problems. These are considered emerging pollutants due to their omnipresence at trace levels in the environment. Nonetheless, the lack of knowledge concerning their impact on the environment, and consequently on aquatic species, highlights this topic as a pivotal concern. Even in low concentrations, antidepressants may cause several effects on the aquatic environment as a result of disturbing homeostasis throughout the central and peripheral nervous systems, both in vertebrates and in vertebrates, and by modifying the regulation of neurotransmitters such as serotonin, norepinephrine and dopamine. Zebrafish (Danio rerio), an aquatic vertebrate species, is one of the most important model organisms in developmental biology, considered extremely valuable for the study of translational neuroscience of complex human brain disorders, being particularly useful for studying genetic and pharmacological mechanisms of depression and antidepressant action.
- Etidronate-based ionic liquids: in vitro effects on bone metabolismPublication . Teixeira, Sónia; Santos, Miguel; Branco, Luís; Fernandes, Maria Helena; Costa-Rodrigues, JoãoThe Bisphosphonates have been used for various purposes since the middle of the 19th century, but the propriety that elicited more curiosity was their ability to regulate bone mineralization. This characteristic made this drug the elite choice for treatments of several pathologies associated to increased bone resorption.
- Uncovering the microglia response during neonatal Group B Streptococcus meningitisPublication . Soares, Joana; Lorga, Inês; Bravo, Joana; Summavielle, Teresa; Nova, Manuel Vila; Bonifácio Andrade, ElvaGroup B Streptococcus (GBS) remains the most common bacterial cause of meningitis in neonates. Microglia, the brain resident immune cells, have a critical role in the development of neural circuits. However, the role of GBS infection on microglia activation and neurological sequelae remains poorly characterised. Here, we aimed to evaluate whether GBS induces changes in microglia profile during the acute phase of infection, using a mouse model that mimics key steps of GBS pathophysiology in humans. Female C57BL/6 mice were intra-vaginally inoculated with GBS during gestation, and CFU analysis was performed on postnatal days (P) 1, 3 and 5. Bacterial colonisation was found at all ages, peaking at P3. When analysing the status of microglia by flow cytometry in the whole brain of male pups at P3, an overall activation was observed in the infected group. Mainly, we found a significant increase in microglia frequency, as well as the mean fluorescence intensities (MFIs) of CD45, CD11b and F4/80. Additionally, we also analysed some microglial receptors that are important neuro-immune regulators with relevant functions during development. We observed increased CX3CR1 expression in microglia, whereas Sirp and CD200r were not altered. Moreover, analysing the cortex and hippocampus, relevant regions for cognition, we found similar numbers in Iba1+ cells, a known microglia marker, in the hippocampus of infected pups. In contrast, a significant decrease was observed in the cortex, suggesting altered migration of these cells. Furthermore, microglia phagocytosis was increased in the cortex of infected pups but not in the hippocampus. Interestingly, quantification of neurons revealed a significant decrease in the hippocampus of infected pups while being increased in the cortex, compared with age-match controls. Altogether, our results show that GBS meningitis alters the neonatal microglia profile. Further studies will be necessary to better understand the microglia inflammatory state after GBS infection.
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