Molecular and Pharmacokinetic Rationale for the Use of Chelidonium majus L. in Wound Healing: An In Silico and In Vitro Validation Ana Borges, Carlos Seiti H. Shiraishi, Rui M. V. Abreu, María Luisa Martín Calvo, Josiana A. Vaz, Ricardo C. Calhelha Molecules, 2026 Wound healing involves the coordinated regulation of inflammation, angiogenesis, and extracellular matrix remodeling, processes modulated by natural bioactives. In this context, Chelidonium majus L. (C. majus), a plant rich in alkaloids and flavonoids, remains mechanistically underexplored. This study, therefore, investigates its metabolites using an integrated computational–experimental approach and evaluates their applicability in sericin-based wound-healing systems. A curated database of 83 C. majus bioactive compounds was analyzed using cheminformatics and molecular docking against key wound-healing targets (iNOS, VEGF, MMP-3, and tyrosinase), followed by ADMET and toxicity prediction (StopTox). Selected plant–sericin formulations were subsequently evaluated for wound-healing activity using an in vitro fibroblast scratch assay. Docking revealed strong binding affinities for several metabolites, particularly protopine, kaempferol-3-rutinoside, cynaroside, hesperidin, quercetin-3-rhamnosylrutinoside, and vitexin, indicating multi-target modulation across inflammatory, proliferative, and remodeling phases of tissue repair. ADMET and toxicity analyses predicted favorable dermal safety and pharmacokinetic profiles for most compounds. Consistently, in vitro assays demonstrated that C. majus–sericin systems had fibroblast migration and wound closure in a concentration- and ratio-dependent manner, with improved healing kinetics observed at 150 µg/mL and for formulations containing higher relative proportions of both components. The experimental outcomes supported the pro-angiogenic and matrix-stabilizing mechanisms predicted in silico. Overall, C. majus metabolites exhibit polypharmacological wound-healing activity, supporting their integration into sericin-based systems as a promising strategy for topical therapies.
Adherence to the Mediterranean Diet and Metabolic Health in Older Adults: Insights from a Feasibility Study Sara Brás Alves, Leandro Moreira de Sá, Carla Agradém, Eugénia Mendes, António Miguel Monteiro, Adília Fernandes, Hélder Fernandes, Josiana Vaz, Ana Pereira Journal of Ageing and Longevity, 2026 Introduction: The Mediterranean Diet is known for its protective effects against cardiovascular and metabolic diseases. Metabolic syndrome, characterized by multiple health risk factors, is increasingly concerning in older populations. Understanding dietary impacts on metabolic health is key for promoting healthy ageing. Objectives: This feasibility study aimed to explore the relationship between adherence to the Mediterranean Diet and metabolic risk factors in older adults participating in a community exercise program and to evaluate the feasibility of applying validated tools in this setting. Methods: A cross-sectional design was used. Adherence to the Mediterranean Diet was evaluated using the PREDIMED questionnaire, while Metabolic Syndrome was evaluated according to National Cholesterol Education Program criteria. Blood samples were taken following WHO guidelines. Results: Ten participants (mean age 73.1 years; 90% women) were included. 50% showed high adherence to the Mediterranean Diet, while 40% had moderate or low adherence. No participants met the full criteria for Metabolic Syndrome. Significant associations were found between Mediterranean Diet adherence and chronic disease (r = 0.869, p < 0.01), and an inverse correlation with the number of Metabolic Syndrome criteria (r = –0.707, p < 0.05). The Mediterranean Diet score was also inversely related to cholesterol (r = –0.740, p < 0.05). Conclusions: Higher adherence to the Mediterranean Diet was associated with better metabolic profiles, highlighting its potential protective role. The study demonstrates the feasibility of incorporating nutritional screening in community exercise programs for older adults. Future research should include larger and longitudinal samples and integrate inflammatory biomarkers.
Antioxidant and Anti-Inflammatory Activities of Probiotic Strains Olga Adriana Caliman-Sturdza, Josiana A. Vaz, Ancuta Veronica Lupaescu, Andrei Lobiuc, Codruta Bran, Roxana Elena Gheorghita International Journal of Molecular Sciences, 2026 This review highlights the anti-inflammatory and antioxidant effects of probiotics and their complex health-related impacts. The main health areas targeted are gastrointestinal inflammation, neuroinflammation, systemic metabolic disorders, and liver conditions. Probiotics work mechanistically to regulate key inflammatory pathways by suppressing nuclear factor (NF-κb) and mitogen-activated protein kinase (MAPK) pathways and activating antioxidant defenses through nuclear erythroid 2-related factor (Nrf2). They stimulate anti-inflammatory cytokines (including interleukin 10 (IL-10) and inhibit pro-inflammatory mediators such as tumor necrosis factor-α (TNF-α), partly through the regulation of T cells. Probiotics also produce antioxidant metabolites (e.g., exopolysaccharides and short-chain fatty acids), which enhance the host’s resistance to oxidative stress. Supplementation with probiotics improves intestinal inflammation and oxidative injury in gut disorders. Clinical trials suggest that probiotic supplements may reduce neuroinflammation and oxidative stress, while improving cognitive or behavioral outcomes in neurodegenerative disorders. Overall, this review underscores that probiotics have potent anti-inflammatory and antioxidant effects within the gut–brain axis and across various organ systems, supporting their use as valuable adjunctive therapies for inflammatory and oxidative stress-related conditions. It further emphasizes that additional mechanistic research and controlled clinical trials are essential to translate these findings into the most effective therapeutic strategies.
Bioactive and Phenolic Profiles in Pinus pinaster Bark: A Comparative Study of Microwave and Ultrasound Extraction Methods Diana Barros, José Ignacio Alonso-Esteban, Tiane C. Finimundy, Carla Pereira, Josiana A. Vaz, Ricardo Pereira-Pinto, Élia Fernandes, Preciosa Pires, Joana Santos, Lillian Barros, Manuela Vaz-Velho Chemengineering, 2026 This study conducted a comprehensive comparison of two green extraction methods, microwave-assisted extraction (MAE) and ultrasound-assisted extraction (UAE), for recovering bioactive phenolic compounds from Pinus pinaster bark. The goal was to valorize timber industry waste and enhance the value of by-products through the development of eco-friendly processes to extract phenolic compounds from Pinus pinaster Aiton subsp. atlantica in northwest Portugal. MAE achieved significantly higher extraction yields than UAE (11.13 vs. 3.47 g extract/100 g bark) and superior total phenolic content (833 vs. 514 mg GAE/g). MAE extracts also exhibited enhanced antioxidant activity in most assays tested (DPPH, ABTS, ORAC, and OxHLIA), while both extracts effectively inhibited lipid peroxidation (TBARS) and showed activity against Gram-positive bacteria. Phenolic profile analysis revealed that MAE recovered a substantially higher amount of total phenolic compounds (230.0 mg/g) compared to UAE (86.95 mg/g), with procyanidins identified as the predominant compounds. The greater recovery of this complex procyanidin mixture by MAE is strongly associated with the enhanced bioactivities observed. Overall, this study confirms MAE as a highly efficient and sustainable technology for transforming pine bark waste into valuable antioxidant and antimicrobial extracts with potential applications in the food and pharmaceutical industries.
Micronucleus assay in buccal and urothelial epithelial cells of wildland firefighters exposed to wildfire smoke Filipa Esteves, Joana Madureira, Bela Barros, Sara Alves, Josiana Vaz, Marta Oliveira, Klara Slezakova, Adília Fernandes, Maria do Carmo Pereira, Simone Morais, Stefano Bonassi, João Paulo Teixeira, Solange Costa Journal of Hazardous Materials, 2025 Occupational exposure as a firefighter is classified as carcinogenic to humans. However, information on the biological effects of wildland firefighting remains limited. This study aimed to assess genotoxicity in a group of wildland firefighters and evaluate the contribution of total concentration of urinary hydroxylated polycyclic aromatic hydrocarbons (ΣOHPAHs) to selected endpoints. A group of 59 northern Portuguese wildland firefighters (mean age: 35.5 ± 9.0 years) was evaluated before and during the wildfire season. Sociodemographic, lifestyle, occupational-related information was collected via questionnaire. The micronucleus (MN) assay in buccal and urothelial cells was applied to paired samples to assess the association with exposure, as indicated by urinary ΣOHPAHs levels. The risk of MN formation in urothelial cells was over twofold higher during the wildfire season [Frequency Ratio (FR): 2.13, 95 % CI: 1.99-2.27, p = 0.01]. A 35 % increase in MN frequency in buccal cells was observed during the wildfire season (FR: 1.35, 95 % CI: 0.76-2.40, p > 0.05). Urinary ΣOHPAHs exhibited a significant positive association with MN‰ in urothelial cells (FR:1.04, 95 % CI: 1.01-1.08, p < 0.05). This study provides the first assessment of MN frequency in urothelial cells of wildland firefighters, offering novel evidence of genotoxic risks and potential long-term health impacts associated with wildland firefighting.
Analysis of the Phenolic Profile of Chelidonium majus L. and Its Combination with Sericin: Balancing Antimicrobial Activity and Cytocompatibility Ana Borges, José Luis Ordóñez-Díaz, Yara Aquino, José Manuel Moreno-Rojas, María Luisa Martín Calvo, Josiana A. Vaz, Ricardo C. Calhelha International Journal of Molecular Sciences, 2025 The incorporation of bioactive natural compounds into biomedical applications offers a promising route to enhance therapeutic efficacy while supporting sustainability. In this study, we investigated the synergistic potential of Sericin, a silk-derived biopolymer, and Chelidonium majus L. (C. majus), a medicinal plant with a diverse phenolic profile, in relation to biological activities relevant for wound care and infection control. A combined experimental strategy was applied, integrating detailed chemical characterization of C. majus extracts with antimicrobial and cytocompatibility assays across different Sericin–plant extract ratios (1:1, 1:2, 2:2, and 2:1). Phytochemical analysis identified and quantified 57 phenolic compounds, including high levels of flavonoids (quercetin, kaempferol, isorhamnetin) and phenolic acids (caffeic and ferulic acid). Salicylic acid (123.6 µg/g), feruloyltyramine (111.8 µg/g), and pinocembrin (98.4 µg/g) were particularly abundant, compounds previously reported to disrupt microbial membranes and impair bacterial viability. These metabolites correlated with the strong antimicrobial activity of C. majus against Gram-positive strains (MIC = 5–10 mg/mL). In combination with Sericin, antimicrobial performance was ratio-dependent, with higher proportions of C. majus (2:1) retaining partial inhibitory effects. Cytocompatibility assays with HFF1 fibroblasts demonstrated low antiproliferative activity across most formulations (GI50 > 400 µg/mL), supporting their potential safety in topical applications. Collectively, the results indicate a concentration-dependent interaction between C. majus phenolics and the Sericin protein matrix, reinforcing their suitability as candidates for natural-based wound healing materials. Importantly, the valorization of Sericin, an underutilized byproduct of the silk industry, together with a widely accessible medicinal plant, underscores the ecological and economic sustainability of this approach. Overall, this work supports the exploration of the development of biomaterials with potential for advancing tissue repair and wound management.
Impact of wildfire emissions exposure on the associations between levels of lung injury, lipid peroxidation, DNA oxidation, and exposure biomarkers Bela Barros, Ana Margarida Paiva, Rui Azevedo, Sara Alves, Filipa Esteves, Adília Fernandes, Josiana Vaz, Maria José Alves, Klara Slezakova, João Paulo Teixeira, Solange Costa, Agostinho Almeida, Marta Oliveira, Simone Morais Science of the Total Environment, 2025 Firefighters face increased risks of developing cardio-respiratory diseases and cancer. This study aimed, for the first time, to simultaneously characterize several biomarkers of effect (lung injury by Clara cell 16 -CC16, lipid peroxidation by 8-isoprostane-8-iso, and DNA oxidation by 8-hydroxy-2-deoxyguanosine-8-OHdG) and exposure (polycyclic aromatic hydrocarbons metabolites - 6 OHPAHs and 17 metal(loid)s) in (pre- and post-exposure) paired urine samples of wildland firefighters, while exploring their inter-/intra-associations and accounting for tobacco consumption. Wildfire combat influenced the levels of CC16 (+39 %), 8-iso (+33 %), 8-OHdG (-13 to +19 %), individual and sum of OHPAHs (+75-211 %), and metal(loid)s (up to 43 %, p > 0.05: lithium, zinc, antimony, and lead); post-exposure increments were more evident among non-smokers. Post-exposure (individual and sum) OHPAHs and some metal(loid)s (copper, cadmium, barium, antimony, copper, lead, zinc, selenium, and rubidium) were positively associated with CC16, 8-iso and/or 8-OHdG (0.609 < r < 0.838; 0.001 < p < 0.047). Spearman's correlations and principal component analysis highlighted CC16 as the best discriminant effect biomarker of wildland firefighting, correlating positively with individual and sum of OHPAHs, cadmium, barium and copper (0.647 < r < 0.764; 0.006 < p < 0.031). Cumulative exposure to wildfires and tobacco contributed to positive correlations (0.587 < r < 0.715; 0.009 < p < 0.045) between lipid peroxidation and arsenic, antimony, lead, and copper, and between DNA oxidation and lead. Smoking firefighters presented higher OHPAHs baseline concentrations (2- to 14-fold), and lung injury and DNA oxidation induced by cadmium, copper, strontium, cesium, barium and thallium (0.661 < r < 0.709; 0.022 < p < 0.038). Given firefighter's carcinogenic risks, performing similar studies in larger groups is crucial to enhance risk assessment by establishing a well-defined panel of effect and exposure biomarkers.
In Silico Identification of Six Mushroom-Derived Sterol and Triterpenoid Compounds as Potential P-Glycoprotein Modulators in Multidrug Resistance Jéssica Fonseca, Carlos S. H. Shiraishi, Rui M. V. Abreu, Sara Ricardo, Josiana A. Vaz Applied Sciences Switzerland, 2025 The overexpression of P-glycoprotein (P-gp) is often directly related to multidrug resistance (MDR), one of the greatest challenges in cancer treatment. This transmembrane efflux pump decreases the intracellular concentrations of chemotherapy drugs, reducing their effectiveness and resulting in treatment failure. This work used in silico methods to assess the potential of bioactive chemicals produced from mushrooms as P-gp modulators. A database comprising 211 bioactive compounds from mushrooms was investigated using molecular docking and virtual screening techniques against the P-gp structure. The compounds ergosta-4,6,8(14),22-tetraen-3-one, lucidumol A, (22E,24S)-ergosta-4,22-dien-3-one, antcin K, 3,11-dioxolanosta-8,24(Z)-diene-26-oic acid, and (22E)-19-norergosta-5,7,9,22-tetraen-3β-ol were identified as the six best candidates from our database of mushroom compounds based on their binding affinities, toxicity predictions, and pharmacological properties assessed through ADME analyses (absorption, distributions, metabolism, and excretion). These six compounds exhibited strong binding affinities, with binding energies ranging from −12.31 kcal/mol to −10.93 kcal/mol, all showing higher affinities than the control, tariquidar, which had a binding energy of −10.78 kcal/mol. Toxicity predictions indicated favorable safety profiles for all six, while ADME analyses found that all six compounds had high oral bioavailability and a low probability of acting as P-gp substrates. These results position bioactive mushroom compounds, particularly these six, as promising P-gp modulators, suggesting positive outcomes in cancer treatment.
Impact of occupational exposure to wildfire events on systemic inflammatory biomarkers in Portuguese wildland firefighters Filipa Esteves, Joana Madureira, Bela Barros, Sara Alves, Joana Pires, Sandra Martins, Marta Oliveira, Josiana Vaz, Klara Slezakova, Maria do Carmo Pereira, Adília Fernandes, Simone Morais, João Tiago Guimarães, Stefano Bonassi, João Paulo Teixeira, Solange Costa Environmental Research, 2025 While occupational exposure as a firefighter is considered a dangerous occupation, research on the underlying mechanisms remains limited, particularly in wildland firefighters. Inflammation, a key effect of wildfire exposure, plays a significant role in the development of various diseases. The current study aims to investigate the impact of wildland firefighting exposure on the levels of pro-inflammatory systemic biomarkers. A pre-post study design investigated 59 wildland firefighters comparing data collected after participation in a wildfire event (Phase II) with data obtained before wildfire season (Phase I). Data on demographics, lifestyle, health and occupational-related factors were assessed. Exposure factors, such as fire combat (e.g., exposure duration), were also registered. Inflammatory biomarkers (i.e. interleukin-6 [IL-6], interleukin-8 [IL-8], tumor necrosis factor α [TNF-α] and high-sensitivity C-reactive protein [hs-CRP]) and hydroxylated polycyclic aromatic hydrocarbons metabolites (1-OHNaph+1-OHAce, 2-OHFlu, 1-OHPhen, 1-OHPyr) were analysed in blood and urine samples, respectively. Serum IL-8 and IL-6 levels were significantly increased after wildland fire combat. IL-8 levels were 2.62 times higher (95 % CI: 1.96-3.50; p < 0.01), whereas IL-6 levels were 1.25 times higher (95 % CI: 1.00-1.57; p = 0.04). Furthermore, IL-8 levels were significantly correlated with urinary 2-hydroxyfluorene levels and fire combat duration (>12 h). In addition, the mean hs-CRP level, in both phases, was above 3.0 mg/L, indicating a potential risk for cardiovascular events. Given the long-term health implications of firefighting occupational exposure, biomonitoring and early detection of occupational risks are essential for protecting firefighters' health. Protective measures must be urgently implemented to enhance occupational health and strengthen preventive strategies in this sector.
Occupational exposure to wildland firefighting and its effects on systemic DNA damage Filipa Esteves, Joana Madureira, Carla Costa, Joana Pires, Bela Barros, Sara Alves, Josiana Vaz, Marta Oliveira, Klara Slezakova, Adília Fernandes, Maria do Carmo Pereira, Simone Morais, Vanessa Valdiglesias, Stefano Bonassi, João Paulo Teixeira, Solange Costa International Journal of Hygiene and Environmental Health, 2025 BACKGROUND: Portugal is among the European Union countries more devastated by forest fires. Wildland firefighters are at the forefront of this battle, facing exposure to a wide range of harmful pollutants. Epidemiological studies have highlighted a potential link between occupational firefighting exposure and several diseases, including cancer. To date, very few studies have explored the biological mechanisms associated with such exposure. The present longitudinal study aims to assess changes in early effect biomarkers following wildland firefighters' occupational exposure to a real wildfire event. METHODS: Paired blood samples from 59 healthy Portuguese wildland firefighters were collected at two different time points: before wildfire season and after a fire event during wildfire season. Sociodemographic variables (e.g., age, sex) and work-related factors (e.g., years of service) were assessed via a self-reported questionnaire. Levels of early effect biomarkers, such as primary DNA damage and oxidative DNA damage (oxidised purines) were assessed via comet assay. DNA double-strand breaks (DSBs) were evaluated by phosphorylated H2AX (γH2AX). Moreover, hydroxylated polycyclic aromatic hydrocarbon metabolites (OHPAHs) and metal(loid)s were quantified in urine samples. The influence of urinary OHPAHs, urinary metal(loid)s, and other exposure-related factors (e.g., firefighting duration) on changes (Δ) in early effect biomarkers (post-vs. baseline levels) was investigated. RESULTS: Firefighting activities led to a significant increase in both primary DNA damage and oxidative DNA damage by 22 % (95 % CI: 1.11-1.35; p < 0.05) and 23 % (95 % CI: 1.04-1.45; p < 0.05), respectively. Results from linear regression revealed that per each unit increase of urinary 2-hydroxyfluorene (2-OHFlu) (μmol/mol creatinine), the risk of ⧍ oxidative DNA damage increased by 20 % [FR: 1.20 (1.09-1.32); p < 0.01]. Additionally, each unit increase in urinary cesium (Cs) (μg/L) resulted in a significant 4 % increase in Δ primary DNA damage [FR: 1.04 (1.01-1.06); p < 0.05] and a 3 % increase in Δ oxidative DNA damage [FR: 1.03 (1.01-1.05); p < 0.05]. Post-exposure levels of γH2AX were significantly correlated with urinary 2-OHFlu levels assessed after firefighting (r = 0.30; p < 0.05). Furthermore, exposure duration and reported breathing difficulties during firefighting were significantly associated with increased levels of primary DNA damage. CONCLUSION: Results obtained provide insights into the potential human health effects of wildland firefighting occupational exposure at the genetic and molecular levels, offering new and important mechanistic data. These findings are crucial for implementing health and safety measures, recommendations, and best practices to mitigate occupational risks and protect the health of wildland firefighters.
The Impact of Sleep on Haematological Parameters in Firefighters Sara Alves, Francisca Silva, Filipa Esteves, Solange Costa, Klara Slezakova, Maria Alves, Maria Pereira, João Teixeira, Simone Morais, Adília Fernandes, Felisbina Queiroga, Josiana Vaz Clocks and Sleep, 2024
Indoor Air Quality at Portuguese Firehouses K. Slezakova, F. Esteves, J. Vaz, S. Costa, M. J. Alves, J. Madureira, B. Barros, A. Fernandes, J. P. Teixeira, S. Morais, M. C. Pereira Studies in Systems Decision and Control, 2024
New trends from fungi secondary metabolism in the pharmaceutical industry Ricardo C. Calhelha, Carlos S. H. Shiraishi, Lúcia Ribeiro, Márcio Carocho, Rui Abreu, Paula Coutinho, Lillian Barros, Josiana Vaz, Isabel C. F. R. Ferreira Natural Secondary Metabolites from Nature Through Science to Industry, 2023
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Firefighter’s occupational exposure to PM2.5 and Polycyclic Aromatic Hydrocarbons Occupational Safety and Hygiene IV Selected Extended and Revised Contributions from the International Symposium Occupational Safety and Hygiene 2016, 2016
Comparative bioequivalence study of rifampicin and isoniazid combinations in healthy volunteers International Journal of Tuberculosis and Lung Disease, 1999
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