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- Book of Abstracts of the 8th Meeting on Medicinal BiotechnologyPublication . Pereira, CassildaO 8º Encontro sobre Biotecnologia Medicinal, é um evento obrigatório para quem se interessa por inovação em saúde e biotecnologia. O encontro reúne pesquisadores, estudantes e profissionais, destacando avanços de última geração e apoiando a colaboração, o networking e o intercâmbio de ideias. Apresentando palestras plenárias inspiradoras de Cláudia Serra (CIIMAR) e Miguel Garrido Costa (Colorifix), ao lado de apresentações orais e de pôsteres, este evento dinâmico da tarde oferece uma oportunidade de se conectar com especialistas e se envolver com os mais recentes avanços científicos em biotecnologia medicinal.
- Biotechnological potential of Macroalgae associated ActinomycetotaPublication . Moreira, Gabriela Alves; Girão, Mariana; Eusébio, Nádia; Mendes, Marta V.; Carvalho, Maria de Fátima; Alves Moreira, GabrielaActinomycetota are Gram-positive bacteria widely distributed across diverse environments, from ter-restrial to marine ecosystems, where they may occur as free-living organisms or in association with other organisms. This phylum is well-known for its remark-able biotechnological potential, with several members producing bioactive compounds such as antimicrobial and anticancer agents. However, macroalgae-associ-ated Actinomycetota remain underexplored despite having a metabolic potential comparable to free-living organisms.The main objective of this study was to explore the biosynthetic potential of 15 Actinomycetota isolates obtained from two marine macroalgae from the Por-tuguese coast, Codium tomentosum and Chondrus cris-pus, through the analysis of their genomes.Total DNA was sequenced using Illumina technology and 17 de novo genomes were assembled. Phyloge-nomic analysis identified 5 different genera, Actinoal-loteichus, Cellulosimicrobium, Kocuria, Nocardia and Streptomyces, with 3 putative novel species.Genome mining, using manually curated antiSMASH data, identified 452 biosynthetic gene clusters (BGCs) across 21 metabolite classes. Among the BGCs iden-tified are antimicrobial and anticancer compounds, siderophores and conserved metabolites. More than 60% of these BGCs have none or low similarity to pre-viously known compounds, which can be an indicator of novelty. Compared with Streptomyces, one of the most studied genera, non-Streptomyces genera exhib-ited a higher novelty percentage (ca. 80%).Additionally, BiG-SCAPE and clinker were used to visualize the data and assess the similarity between BGCs for the same compound in different strains.Overall, these results confirm that macroalgae-as-sociated Actinomycetota represent a valuable and underexplored source of new species and biosynthet-ic diversity, supporting further study of this bacterial group.
- Betulinic acid modulates adipocyte differentiation and secretome activity: Downstream effects on Glioblastoma cell migrationPublication . Gouveia, Afonso; Ferraz, Ricardo; Prudêncio, Cristina; Fernandes, Silvia; Fernandes, SílviaBetulinic acid (BA) has been recognized for its po-tential to modulate tumor progression and adipocyte function, yet its effects on adipocyte secretome re-modeling and the downstream consequences for glio-blastoma behavior remain incompletely understood1-4. In this study, we investigated the influence of BA on adipocyte secretory activity and its functional impact on glioblastoma cell migration. Direct exposure of U251 glioblastoma cells to BA significantly reduced cell migration, confirming its intrinsic anti-migratory potential. To further explore microenvironmental in-teractions, adipocyte-conditioned media (ACM) de-rived from BA-treated adipocytes was evaluated for its effects on glioblastoma behavior. Secretome anal-ysis demonstrated that BA altered matrix metallopro-teinase (MMP) expression and reduced gelatinolytic activity, indicating significant modulation of extracel-lular matrix–related proteolytic pathways. Functional assessment using wound healing assays revealed that ACM from BA-treated adipocytes did not consistent-ly suppress glioblastoma migration. Intermediate BA concentrations were associated with increased migratory behavior relative to basal conditions, although differences compared with control ACM were not statistically significant. Additionally, transwell migra-tion assays did not demonstrate significant differ-ences between treatment conditions, suggesting that BA-induced alterations in adipocyte secretome com-position do not uniformly translate into measurable changes in glioblastoma migratory capacity across dif-ferent experimental models. These findings indicate that modulation of gelatinolytic pathways alone is insufficient to predict glioblastoma cell migration and support the contribution of additional adipocyte-de-rived signalling mechanisms in regulating tumor be-havior. Overall, BA induces substantial remodelling of adipocyte secretory function, producing complex and non-linear downstream effects on glioblastoma cells. This study highlights the importance of tumor micro-environment crosstalk and underscores the need to evaluate multiple regulatory pathways when consid-ering the therapeutic potential of BA and related bio-active compounds.
- Enhancing antifungal activity against Candida albicans using novel organic compoundsPublication . Alves, Beatriz; Silva, Rui; Vieira, Mónica; Ferraz, Ricardo; Fernandes, Silvia; Teixeira, Vitor; Pereira, Clara; Almeida Vieira, Mónica Andreia; Fernandes, SílviaCandida albicans is the predominant cause of inva-sive candidiasis, a major global health threat [1 The increasing emergence of resistance to commonly used antifungals (ex.:iazoles and polyenes) demonstrates an urgent need for alternative therapeutic strategies. The objective of this study was to evaluate the antifungal activity of novel organic derivatives from bioactive natural compound, including Naphthalene, Betulinic Acid (BA) [2]; Protocatechuic Acid (PCA) and Gallic Acid (GA), alone, or in combination with conventional antifungals to investigate synergistic effects that en-hance their efficacy. Antifungal susceptibility assays were performed in C. albicans ATCC 10231 using the broth microdilution method, according to CLSI M27 guidelines, to determine Minimum Inhibitory Concen-trations (MICs) and Minimum Fungicidal Concentra-tions (MFC). The interaction between BA derivatives and amphotericin B was assessed by checkerboard assay and fractional inhibitory concentration index (FICI). Naphthalene, PCA and GA derivatives showed low antifungal activity, falling within the moderate to weak bioactivity range (MIC ≥ 100 μg mL−1; [3]). Among BA derivatives, BA-4 and BA-5 exhibited the best antifungal activity, with MIC values of 0,95 and 1,91 μg mL−1, respectively, achieving 99% fungicidal activity at the same concentration. Furthermore, in combination assays, both compounds enhanced Am-photericin B activity, showing synergistic effects (FICI = 0.50), resulting in a decrease in MIC. Although most newly synthesized compounds exhibited limited anti-fungal potential, BA-4 and BA-5 demonstrated very strong bioactivity and fungicidal effects against C. albicans. The observed synergy with amphotericin B highlights their potential role as antifungal adjuvants and supports the further exploration of betulinic acid derivatives as viable approaches to overcoming anti-fungal resistance.
- Profiling airway microbiome composition through volatilomicsPublication . Neto, Filipa; Ferraz, Ricardo; Vieira, Mónica; Prudêncio, Cristina; Rufo, João; Almeida Vieira, Mónica Andreia; Cavaleiro Rufo, JoãoThe airway microbiome is known to mediate respi-ratory health. However, current available methods for microbiome analysis are time-consuming and/or rep-resent significant costs for a generalized application in clinical practice. Volatilomics has been suggested as a rapid and low-cost approach to screen microbial pro-files in human samples. Therefore, we aimed to study the efficacy of volatilomics in discriminating microbi-al isolates collected from human breath condensate samples. Bacterial strains showing significant growth under conditions simulating the respiratory environ-ment were isolated. Each strain was standardised to an inoculum of 10⁸ CFU/mL and analysed using an elec-tronic nose equipped with a six-sensor matrix. Data were explored through principal component analysis, cluster analysis, pattern analysis, random forests and recursive partitioning regression. One sensor was re-moved from the analysis due to high collinearity. Prin-cipal component analysis was able to separate strains and the control mainly through the second principal component (p = 0.024), characterized by high MQ3 and MQ8 sensor responses. Sensor profile maps showed distinct volatile patterns across strains (Fig-ure 1), suggesting the presence of distinct microbial signatures. However, reproducibility was low between replicas and time since culture. Recursive partitioning for separating sterile controls from inoculated sam-ples showed the highest accuracy (AUC = 0.73). These results show the potential of separation of microbial strains based on volatilomics. Nonetheless, relative robustness was only achieved for the discrimination of sterile vs inoculated samples.
- Impact of simulated digestion on microplastics: Surface changes and intestinal in vitro exposurePublication . Lamas, Mariana; Fernandes, Virgínia Cruz; Oliveira, Marta; Rodrigues, Francisca; Fernandes, VirgíniaMicroplastics (MP) have been widely detected in a variety of food products1, and ingestion is one of the main pathways for human exposure to these contam-inants. Once ingested, MP might interact with the gastrointestinal (GI) environment, yet it remains un-clear whether digestive processes alter their chemical structure or mainly influence their surface properties, and possibly their interaction capacity3. This study in-vestigates digested-induced modifications in MP and establishes suitable conditions for intestinal in vitroassays. Simulated digestion of six polymers (PTFE; PMMA, PA6, uPVC, PET, and LDPE) followed the IN-FOGEST model2. Following digestion, particles were analysed using Fourier-transform infrared spectros-copy (FTIR) to assess structural and surface changes. Cell viability was evaluated, by MTT, in Caco-2 and HT29-MTX intestinal cell lines exposed to different dilutions (1:10, 1:20, and 1:40) of digestion fluids, with respective MP. FTIR results indicated that all polymers maintained their fundamental chemical structure after digestion. Instead, consistent surface-related alter-ations were observed, including reduced intensity of characteristic polymer bands and enhanced signals in regions associated with proteins and hydroxyl groups, suggesting biomolecules adsorption and increased hy-dration. Cell studies demonstrated that 1:20 dilution of digestive fluids with MP maintained the cell viability above 70%, representing the most suitable condition tested. Simulated GI digestion appears to preserve the intrinsic chemical identity of MP, while promoting surface modifications that may affect their biological interactions. The identification of 1:20 digestion fluid dilution supports its application in future studies using advanced intestinal models, including three-dimen-sional systems and assessment of cellular oxidative stress responses.
- Bioactive contact lens incorporating umbilical cord-derived Mesenchymal Stem Cell Secretome for corneal regenerationPublication . Coutinho, Ana; Sousa, Ana Catarina; Maurício, Ana ColetteCorneal injuries and degenerative corneal disorders remain a significant clinical challenge, as impaired tis-sue regeneration can lead to corneal opacity, neovas-cularization, and progressive vision loss [1,2]. Current therapeutic approaches, including topical eye drops and corneal transplantation, are limited by poor bio-availability, insufficient regenerative potential, and the risk of immune rejection [3]. Therefore, there is a pressing need for innovative, non-invasive strategies capable of restoring corneal function. In this context, conditioned medium derived from umbilical cord mes-enchymal stem cells (UC-MSCs), rich in bioactive fac-tors (secretome), has shown promising regenerative and immunomodulatory effects in corneal repair. This study aims to develop a bioactive contact lens as a non-invasive therapeutic platform for corneal regener-ation. The system is based on a hydrogel composed of polyvinyl alcohol (PVA), hyaluronic acid (HA), and UC-MSC-derived conditioned medium. Prior to incorpo-ration, the secretome will be characterized to identify key biomarkers associated with immunomodulation and tissue regeneration and validated in vitro through cytocompatibility and cell migration (scratch) assays. The bioactive contact lenses will be fabricated using mold-based techniques to ensure reproducible geom-etry and structural consistency. Within the hydrogel matrix, PVA provides mechanical stability and flexi-bility, while HA enhances hydration, biocompatibility, and mimics native corneal properties, contributing to comfort and biological performance. This platform enables sustained and localized delivery of bioactive factors directly to the injured corneal surface. The re-sulting hydrogel will undergo physicochemical charac-terization using Fourier Transform Infrared Spectros-copy (FTIR) and Scanning Electron Microscopy (SEM). Biological performance will be evaluated in vitro using corneal epithelial cells under conditions that mimic the ocular environment. Overall, this work proposes a promising cell-free regenerative strategy for corneal wound healing, with the potential to enhance epitheli-al repair, reduce fibrosis, and improve visual outcomes.
- Glioma behavior under the influence of Cinnamic Acid: a cellular approachPublication . Moreira, Ana; Almeida, Joana; Ferraz, Ricardo; Ferraz, RicardoCancer is one of the biggest health problems worldwide, characterized by uncontrolled proliferation, rapid growth and the ability to invade other tissues (1). Gliomas are highly challenging tu-mours from a therapeutic perspective, so new and more effective therapeutic strategies have been in-vestigated, with particular focus on the therapeu-tic potential of plant-derived compounds, such as Cinnamic Acid (2,3). Cinnamic Acid, extracted from Cinnamomum cassia, has proved to be a promising therapeutic agent, as it exhibits antitumor properties and low cytotoxicity toward healthy cells (4–6). This phenolic compound has the ability to cross the blood-brain barrier and affect tumour cell survival by reduc-ing their proliferation rate, inducing apoptosis and inhibiting key signalling pathways involved in tumour growth (7–9). This study aims to evaluate the glioma behaviour under the influence of cinnamic acid, using a GL261 cell model. To this end, cells were cultured and treated with different concentrations of cinnamic acid (1x10-3 mol/L to 4x10-3 mol/L), followed by in vi-tro assays to assess its impact on cell viability and cell migration. The MTT assay performed on the GL261 cell line demonstrated that Cinnamic Acid is capable of reducing cell viability at all tested concentrations, with a cytotoxic effect that tends to be dose-dependent. At lower concentrations (1x10-3 mol/L and 2x10-3 mol/L), cell viability was approximately 83% (SD=13,3% and 13,9%, respectively) and the most pronounced reduc-tion in cell viability was detected at the highest tested concentration (4x10-3 mol/L), with a viability of 76% ± 13,9%, corresponding to an approximately 24% re-duction relative to the control (viability of 100%).
- Evaluation of the concordance of pharmacogenomic information in the RCM and FI with the international recommendations of CPIC and PharmGKBPublication . Almeida, Nuno; Vasconcelos, Ana; Lacerda, Ivo; Santos, Marlene; Santos, MarlenePharmacogenomics plays an increas-ing role in optimizing antiviral therapy, allowing the choice and dosage of drugs to be tailored to the pa-tient’s genetic profile. However, the integration of this information into regulatory documents, such as the Summary of Product Characteristics (SmPC) and the Patient Information Leaflet (PIL), remains inconsistent. To evaluate the degree of concordance between the pharmacogenomic information present-ed in the SmPC and PIL of antiviral drugs with the international recommendations of the Clinical Phar-macogenetics Implementation Consortium (CPIC) and the evidence levels reported in PharmGKB. An analysis was conducted on a set of 146 marketed antiviral drugs, assessing the presence of pharmacogenomic recommendations in their respec-tive SmPCs and PILs. These data were compared with CPIC guidelines and the pharmacogenomic evidence levels assigned by PharmGKB. Results: Only 14 out of the 146 drugs analysed pre-sented recommendations based on CPIC. Regarding PharmGKB evidence levels, most active substanc-es exhibited level 3. Complete agreement with CPIC guidelines was observed in only 2 SmPCs, while other 5 did not consider any genetic polymorphisms. The re-maining documents contained incomplete or partially aligned pharmacogenomic information relative to in-ternational recommendations. The results show that, with the excep-tion of abacavir, there is a discrepancy between the available pharmacogenomic evidence and its inte-gration into the SmPCs of antiviral medicines. This misalignment reinforces the need for more compre-hensive studies and effective harmonization between international guidelines and the content of SmPCs, promoting the adoption of personalized medicine.
- Optimization of SPE-HPLC-FLD methodologies for the extraction and detection of steroid hormones in plasmaPublication . Bracchi, Isabella; Paíga, Paula; Delerue-Matos, Cristina; Pestana, Diogo; Negrão, Rita; Keating, Elisa; Fernandes, VirgíniaThe quantification of steroid hormones, such as es-tradiol and ethinylestradiol, in plasma requires sensi-tive and reliable analytical methodologies1. This study presents the optimization of a bioanalytical method based on Solid-Phase Extraction (SPE) coupled with High-Performance Liquid Chromatography with Flu-orescence Detection (HPLC-FLD). For sample ex-traction, Strata-X (polymeric reversed-phase) cartridg-es were used, where critical SPE parameters, including cartridge conditioning, matrix washing, and analyte elution, were evaluated. Chromatographic separation was achieved on a C18 stationary phase using a mobile phase of acetonitrile and 0.1% formic acid in ultrapure water. Both isocratic and gradient elution programs were tested to maximize resolution and peak shape. The method was validated for linearity, recovery, and precision (intra- and inter-day), meeting all established criteria for bioanalytical applications. This combined optimization resulted in improved selectivity and sen-sitivity for the determination of estradiol and ethinyl-estradiol in plasma (Figure 1), demonstrating a robust bioanalytical tool for monitoring these compounds in complex biological matrices.
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