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- The synergy of dopaminergic system and adult hippocampal neurogenesis in a pre-clinical model of Parkinson’s disease pp85Publication . Araújo, B.; Caridade-Silva, Rita; Vilaça-Ferreira, A.; Martins-Macedo, J.; Teixeira, C.; Soares-Guedes, C.; Svenningsson, P.; Pinto, L.; Teixeira, F.; Guedes, CarlaDepressive disturbances are prevalent in 40% to 50% of clinical cases of Parkinson’s Disease (PD), alongside a common reduction in adult hippocampal neurogenesis observed in both PD and its related conditions. This neurogenesis deficit may affect the clinical course of the disease. With this in mind, we set an experiment using the glial fibrillary acidic protein-thymidine kinase (GFAP-TK) transgenic rat model to assess the impact of impaired adult cytogenesis induced by the antiviral Ganciclovir on PD. The experiment involved a combination of the GFAP-TK model and a 6-hydroxydopamine (6-OHDA) model of PD, while behavioral analyses focused on anxiety, depression, and motor skills. From the results, histological examinations revealed decreased proliferative cells and reduced dopaminergic innervation. Additionally, analysis of newborn and immature neurons occurred in the hippocampus, subventricular zone, and olfactory bulbs, while dopaminergic loss was assessed in regions like the substantia nigra and striatum. Findings indicated that the model exhibited anxiety/depressive-like behaviors and motor impairments, linked to the notable loss of dopaminergic neurons, which appeared to correlate with reduced doublecortin-positive cells in the hippocampus. Moreover, results suggested subtle differences between ipsilateral and contralateral sides, highlighting the dopaminergic system's role in hippocampal adaptation. Therefore, these findings suggest a connection between reduced neurogenesis and dopaminergic neuron loss, hinting that these phenomena might be interrelated. Therefore, investigating this potential regional interconnection may augment our understanding of non-motor dimensions in PD pathophysiology related to motor functions, thereby facilitating the development of enhanced therapeutic strategies for individuals in the early stages of PD.
- Beyond the brain: The hidden role of cardiorenal dysfunction in Parkinson’s diseasePublication . Teixeira, C.; Araújo, B.; Caridade-Silva, Rita; Martins-Macedo, J.; Guedes,Carla; Gomes, Eduardo; Falcão-Pires, I.; Alencastre, I.; Teixeira, F.; Guedes, Carla; Gomes, EduardoParkinson’s disease (PD) is the second most common neurodegenerative disorder, marked by the progressive loss of dopaminergic neurons in critical areas of the brain, particularly the striatum and substantia nigra. PD's complex nature suggests its interactions with various systemic health issues, particularly those affecting organs outside the central nervous system (CNS), which may increase the risk of developing PD and affect treatment outcomes. Research indicates that individuals with cardiovascular disease (CVD) and chronic kidney disease (CKD) face significantly higher risks of PD, even when controlling for shared risk factors. Notably, alpha-synuclein aggregations, a hallmark of PD, have also been found in the renal and cardiac tissues of patients with PD, CKD, and CVD, highlighting the interconnectedness of these systems. The Zucker fatty and spontaneously hypertensive (ZSF1) rats model metabolic syndrome, which includes kidney issues and heart failure. This study aimed to explore how the ZSF1 phenotype impacts the integrity of dopaminergic neurons and neuroinflammatory processes. Brain tissues from ZSF1 rats were analyzed through immunostaining with markers specific to dopaminergic and glial cells. The results showed a significant decrease in dopaminergic markers in the striatum and substantia nigra, indicating a potential link between cardiorenal dysfunction and neurodegenerative pathways. These findings suggest that systemic health conditions can directly influence PD pathology, emphasizing the complex interactions between the brain, heart, and kidneys, and presenting new opportunities for targeted PD therapies.
- Exploring the bioactive potential of Gracilaria gracilis: An extraction optimization study using response surface methodologyPublication . Neves, Cátia; Morais, Stephanie; Vale, Tiago; Soares, Cristina; Grosso, Clara; Domingues, Valentina F.; Vieira, Mónica; Delerue-Matos, Cristina; Vieira, MónicaThe extraction of bioactive compounds from the seaweed Gracilaria gracilis was optimized for food use using a Response Surface Methodology. Two designs, Central Composite Face-centered (CCD) and Box-Behnken (BBD) assessed the effects of extraction time, temperature, and seaweed-to-solvent ratio using water as the solvent. The extraction yield was assessed by Total Phenolic Content (TPC). BBD’s best model was a Reduced Quadratic (R2 = 0.9356), predicting 3.336 mg GAE/L at 74 °C in 1.4 h with a 1:75 ratio. CCD’s top was Reduced Cubic (R2 = 0.9091), forecasting 4.278 mg GAE/L at 46 °C in 1.1 h, the same ratio. The actual obtained TPC values were 4.35 mg GAE/L for BBD and 4.25 mg GAE/L for CCD.
- Utilizing spent yeast for tannin adsorption in chestnut shell treatment solutionsPublication . Vieira, Elsa F.; Amaral, Tomás; Ferraz, Ricardo; Delerue-Matos, Cristina; Ferraz, RicardoThis study evaluated the use of brewer’s spent yeast (BSY) as an adsorbent for tannins from a chestnut shell extract (CS tannin extract). This extract was derived from an alkaline treatment (5% NaOH (v/v)) to recover cellulosic material from chestnut shells and needed valorization. Various BSY treatments, including lyophilization, immobilization in calcium alginate beads, and alkaline and acid treatments, were tested to identify which had the best tannin adsorption capacity. The results highlight BSY’s potential as a system to valorize tannins from this treatment solution.
- Deciphering the influence of CD276 glycocode in gastric cancer immunoregulationPublication . Magalhães, Mariana; Dourado-Gonçalves, Martina; Santos, Lúcio Lara; Peixoto, Andreia; Ferreira, José AlexandreGastric cancer (GC) ranks 5th globally in incidence and cancer-related mortality, with advanced-stage 5-year survival rates below 20%1,2, unveiling the need for novel immunotherapies. CD276, an immune checkpoint molecule, is related to immune evasion and tumour progression in GC3-5. In turn, aberrant glycosylation, especially truncated O-glycans, Tn and Sialyl-Tn, correlates with GC aggressiveness and poor prognosis6. Its clinical impact relies on the modulation of protein function and immune escape5,7. This study hypothesizes that CD276 glycosylation may offer new targets for immunotherapy in GC. Methods: CD276 expression was assessed in AGS and MKN-45 cell lines using qPCR and immunoblotting. Glycoengineered AGS and MKN-45 cell lines with truncated glycosylation (C1GALT1 KO) and wild-type (WT) controls were analysed similarly. In vitro proliferation, migration and invasion assays were conducted. CD276 expression was assessed in AGS and MKN-45 cell lines using qPCR and immunoblotting. Glycoengineered AGS and MKN-45 cell lines with truncated glycosylation (C1GALT1 KO) and wild-type (WT) controls were analysed similarly. In vitro proliferation, migration and invasion assays were conducted. Both cell lines have CD276 expression in WT cells. CRISPR/Cas9-mediated knockout of C1GALT1 did not significantly alter CD276 protein or mRNA expression levels. The proliferation assay did not show any changes in either cell line. However, migratory and invasive assays promoted a cell line-dependent response. The migratory capacity of AGS decreased with truncated glycosylation while increasing for MKN-45. C1GALT1 KO AGS cells showed enhanced invasiveness. Aberrant glycosylation in AGS and MKN-45 cells did not affect CD276 expression in vitro, suggesting a CD276 expression independent of O-glycan elongation in GC. Tn/STn glycophenotypes have a cell line-dependent functional impact in GC, suggesting glycosylation-related migration and invasion capacities. Future proteomic studies will clarify the results. Moreover, further research will identify CD276 glycoproteoforms and assess CD276 abrogation effects on T-cell immunomodulation and cytokine profiles. This research may contribute to the development of novel targeted therapies and improved patient outcomes in GC.
- MicroRNA-mediated control of mesenchymal stem/stromal cell senescence: A comparative in vitro studyPublication . Giestinhas, Andrea; Moura, Sara Reis; Silva, Elsa; Nascimento, Diana; Almeida, Maria InêsOsteoarthritis is a growing health problem, aggravated by aging. Mesenchymal stem/stromal cells (MSCs) emerged as a promising cell-based therapy for this disease. However, in vitro expansion of MSCs can result in cell senescence. MicroRNAs (miRNAs) are key regulators of senescence pathways, making them attractive targets to decrease senescence. This work aims to identify senescence-related miRNAs, with the long-term goal to optimize large-scale MSC expansion. Human primary MSCs from 30-60 years patients were isolated and characterized. Senescence was induced by exposing MSCs to 5, 10 and 20 Gy of gamma radiation or to 200, 400, 600 μM of hydrogen peroxide (H2O2). Senescence was confirmed by β-Galactosidase staining along with fluorescence immunocytochemistry to assess Ki67 (proliferation marker) and γH2AX (DNA damage marker). Senescence associated with miRNAs were analyzed by RT-qPCR. Statistical analysis was performed using GraphPad Prism 8.0 (P-value<0.05 was considered statistically significant). Radiation successfully induced senescence in MSCs, particularly in the 20 Gy condition compared to the control. Specifically, γH2AX increased 60%, Ki67 decreased 25%, and β-Gal levels were increased. miR-29 family and miR-34a were upregulated at 20Gy compared to the control, while no differences were found for miR-16, miR-195, and miR-126. On the other hand, neither of the tested H2O2 concentrations showed significant differences in inducing senescence in MSCs. Our data shows that gamma radiation can be used as an in vitro model to induce MSC senescence and to identify senescence-associated miRNAs, while single treatments with H2O2 do not significantly impact senescence.
- Uncovering the mechanisms of protection during mycobacterium tuberculosis infectionPublication . Queirós, Sofia; Silva, Marta L.; Fernandes, Ana I.; Medina-Lopes, Mónica; Saraiva, Margarida; Cardoso, Marcos SantosTuberculosis (TB) is one of the most devastating infectious diseases affecting humankind, having caused 1.25 million deaths in 2023. The identification of effective protective mechanisms in TB could critically contribute to the urgent need of new therapies to stop TB. It is known that genetically related Mycobacterium tuberculosis (Mtb) isolates shape the immune response and disease outcomes. Using the natural genetic diversity of Mtb as a tool to understand protection in TB, my project focuses on the slow progressing infection caused by Mtb isolates of lineage 6 (L6), in which we hypothesize that critical protective mechanisms operate. Analyses of in vitro (macrophages) and in vivo (mouse) infections were performed in susceptible C3HeB/FeJ or resistant C57BL/6 mice, to compare the immune response and outcome of infection caused by Mtb belonging to L6 or L4. Results: In vitro (macrophage) infections showed that both Mtb L6 and Mtb L4 isolates grow overtime, but Mtb L6 grows slower than Mtb L4. In vivo, we found that C57BL/6 mice infected with Mtb L6 offer a unique model of full protection. Strikingly, flow cytometry analysis revealed a faster recruitment of activated CD4+ T cells to the lungs in the Mtb L6-infected C57BL/6, which might suggest that protection may be achieved by this early T cells recruitment. Our results indicate that a slower growth of Mtb L6 in macrophages together with earlier T cell responses in the lung may underlie fully infection control. The ongoing RNA-sequencing analyses will highlight putative protective immune mechanisms operating in this model, of great interest to devise new strategies to control TB.
- Copine 1 counteracts pneumolysin-induced plasma membrane damage caused by Streptococcus pneumoniae infectionPublication . Monteiro, João; Lima, Ricardo; Vale-Costa, Sílvia; Sousa, SandraStreptococcus pneumoniae causes pneumonia, meningitis, and sepsis, leading to millions of hospitalizations and deaths annually despite vaccination efforts. Understanding bacterial interaction with host cells is key to developing innovative therapies. Pneumolysin (PLY), a S. pneumoniae virulence factor, forms pores in the host cell plasma membrane (PM) to disrupt cellular integrity and promote bacterial dissemination. Host cells rely on repair mechanisms to counteract PM damage induced by PLY(1). We recently identified Copine 1, a calcium-dependent phospholipid-binding protein, as a key player in this repair process(2) However, its precise role in PM repair remains unclear, which is the focus of this study. To ascertain if Copine 1 redistributed to PM damage sites, A549 (lung epithelial) or HeLa (cervix epithelial) cells were treated with purified PLY, infected with S. pneumoniae, or untreated. Protein localization was assessed by immunofluorescence confocal microscopy. To identify Copine 1 partners involved in PM repair, HEK 293T (kidney epithelial) cells expressing Copine 1 fused to a biotin ligase were non-intoxicated or intoxicated with PLY in the presence of exogenous biotin. Biotinylated proteins were purified and identified by mass-spectrometry. Following both PLY intoxication and bacterial infection, Copine 1 was recruited to nonmuscle myosin heavy chain IIA protein accumulations at the PM, which are PM sites associated with effective repair upon damage by other bacterial pore forming toxins (3,4). The results from the mass-spectrometry analysis are pending. These findings suggest, thus far, that Copine 1 counteracts PLY-mediated PM damage by participating in cortical actomyosin cytoskeleton remodeling.
- A practical case of mentoring in higher education at a health schoolPublication . Saúde, Miguel; Ferraz, Ricardo; Silva, Vítor; Barreto, João; Portugal, Paula; Santos, Joana; Vieira, Mónica; Vieira, Mónica; Santos, Joana; Barreto, João Francisco; Simões-Silva, VitorThe rapid advancement of Generative Artificial Intelligence (AI) tools is reshaping higher education, particularly in creative fields such as design, multimedia, and audiovisuals. Students increasingly seek faster solutions for academic challenges, often relying on AI-generated content rather than engaging in research, critical analysis, and validation through credible sources or instructors. This tendency weakens the "thinking vs. creating" stage of the design methodology, a fundamental cognitive process that requires deep concentration, research, and creative reasoning to materialize ideas effectively. The proposed design methodology consists of four stages: (1) Briefing—problem definition, (2) Thinking vs. Creating—ideation, (3) Materialization—execution based on Lupton’s (2016) Design Thinking principles, and (4) Solution—evaluating originality, relevance, and creativity. In practical-laboratory classroom settings, students engage in manual techniques such as mind maps and moodboards (A2 format). Initially, they hesitate to move away from digital tools but later recognize the cognitive and creative benefits of these techniques. This process reinforces the understanding that strong ideas require effort rather than instant AI-generated solutions. A survey had a total of 256 answers of students in Communication Design, Multimedia, and Audiovisuals, revealed that 64.8% use ChatGPT, 28.9% Capcut, 19.5% Copilot, 13.3% Adobe Firefly, 11.7% DALL·E, and other AI tools. However, only 41.4% critically reflect on AI-generated content, 17.6% validate it with teachers, and 14.5% verify information through online sources. This study concludes that while AI accelerates and optimizes aspects of the creative process, human cognitive engagement remains essential. The integration of AI in co-creation processes, particularly at the "thinking vs. creating" stage, enhances skill development when used as a complementary tool rather than a replacement for critical and creative thinking.
- Goji berries (Lycium barbarum) extract as new potential cosmetic ingredient – A first screeningPublication . Silva, Joana; Teixeira, Filipa; Silva, Ana M.; Vieira, Mónica; Amaral, M. Helena; Rodrigues, Francisca; Vieira, MónicaCurrently there is greater concern about appearance and skin care products. Although cosmetic products are not associated with serious health problems, there are concerns related to undesirable effects due to the presence of potentially carcinogenic and allergenic ingredients. Thus, consumers have been increasingly looking for products of natural and plant-based origin. The use of these products is expected to increase exponentially, and studies are urgently needed to introduce new products into the market. Goji berry is a fruit with increasing popularity due to its richness in phenolic compounds, carotenoids, organic acids, carbohydrates, and vitamins. Its antioxidant, antimicrobial and anticancer activities make this product promising for cosmetic applications. The aim of this study was to study an ecological extract obtained from goji berries through ultrasound-assisted extraction as a new ingredient incorporated into a cosmetic formulation. Goji berry extract was obtained by ultrasound-assisted extraction, according to Silva et al. . To assess safety, in vitro assays were carried out on skin cell lines. Subsequently, several cosmetic formulations incorporating the extract were developed and characterized. The results demonstrated that goji berries have no toxicity on skin cell lines. Cosmetic formulations containing the goji berries extract demonstrated to be suitable for application to the skin and maintained their characteristics during the storage period. Goji berries extract is safe and stable in cosmetic formulations, proving to be suitable for consumer needs.