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- Propolis-Loaded Niosomes for Dermopharmaceutic and Cosmetic Applications: Development, Stability, Safety, and In Vitro Bioactivity StudiesPublication . Pinto, Maria Beatriz; Pires, Patrícia C.; Correia, Mafalda; Moço, Gabriela; Sousa-Oliveira, Inês; Sousa, Maria João; Calhelha, Ricardo C.; Vilas-Boas, Miguel; Falcão, Soraia; Mazzola, Priscila Gava; Veiga, Francisco; Paiva-Santos, Ana CláudiaPropolis, produced by Apis melliferabees, is composed of several biologically relevant phenolic compounds with known analgesic, anti-inflammatory, antitumor, antioxidant, immunomodulatory, wound healing, and antibacterial effects, having gained significant interest for therapeutic and cosmetic purposes. Niosomes, self-assembled vesicular nanosystems, are highly researched for topical delivery due to providing controlled and sustained release, protecting encapsulated compounds from degradation, improving stability, and having good biocompatibility and biodegradability. This work aimed to develop novel propolis-loaded niosomes with small and homogeneous particle size, high encapsulation efficiency, controlled release, adequate stability, relevant bioactivity, and high safety for topical application, for therapeutic and/or cosmetic purposes. Aided by quality by design (QbD) analysis, niosomes containing Tween 20, Kolliphor RH 40, cetyl alcohol, and/or cholesterol were produced by thin-film hydration followed by extrusion, with small particle size (between 100 and 200 nm), homogeneous distribution (PDI below 0.2), relevant zeta-potential (around -38 mV), good stability both under refrigeration and at room temperature, high encapsulation efficiency (ranging from 78.8 to 87.4%), and a controlled release profile, relevant to ensure prolonged bioactivity at the application site. Adequate concentration-dependent in vitro safety in keratinocytes and fibroblasts (up to 12.5 or 25 mu g/mL) was demonstrated as well, with some niosomal formulations also showing a low irritative potential in a HET-CAM test, and relevant in vitro anti-inflammatory potential (IC50 from 14.90 to 17.89 ng/mL). These novel nanoplatforms, containing a nature-derived hydrophobic compound with various relevant bioactivities, could serve as versatile and advantageous formulations in both pharmaceutical and cosmetic contexts of application.
- Anthropogenic Barriers Limit Fish Access to Essential Habitats in the Amazon in the Face of Climate ChangePublication . Yofukuji, Kátia Yasuko; Fabrin, Thomaz Mansini Carrenho; Stabile, Bruno Henrique Mioto; Agostinho, Angelo Antonio; Jézéquel, Céline; Batista‐Silva, Valéria Flávia; Esser, Luiz Fernando; Ferreira, José Hilário Delconte; Ré, Reginaldo; Tedesco, Pablo A.; Azevedo, João C.; Bailly, DayaniBarriers represent one of the greatest threats to river integrity and freshwater fish, as they fragment habitats and impair species dispersal, particularly in a scenario of climate change. In this context, we applied a novel framework that combined predictions of species distribution models with a river connectivity index to identify accessible and climatic‐environmental suitable habitats for frugivorous and socioeconomically important fish in the Amazon basin. We also ranked dams based on their potential for river fragmentation and blocking access to climate refuge for fish species that provide essential ecosystem functions and services in the Amazon. Our results revealed that there are still extensive areas that remain both connected and climatic‐environmentally suitable along the Amazon‐Solimões rivers, acting as core areas for fish dispersal and tracking suitable habitats. However, the planned expansion of hydropower infrastructure combined with climate change can lead to a contraction of areas that will remain simultaneously climatic‐environmental suitable and connected. By identifying and ranking the most impactful barriers, our results can provide innovative and applicable information for sustainable energy planning decisions in the Amazon. These results can inform policies and conservation actions aimed at preserving river connectivity, biodiversity, and ecosystem services under rapidly changing conditions.
- Use of Permeable Reactive Barriers in the Removal of ACT and DCF from Effluents of Wastewater Treatment PlantsPublication . Kalmakhanova, Marzhan S.; Khabashova, Aidana U.; Nurlybayeva, Aisha N.; Orynbayev, Seitzhan A.; Gomes, Helder; Snow, Daniel D.Pharmaceuticals such as paracetamol and diclofenac (DCF) are among the most extensively consumed drugs worldwide and are continuously released into municipal and hospital wastewater due to incomplete human metabolism. Their persistent presence in aquatic environments, typically ranging from ng/L to µg/L, raises concerns due to endocrine disruption, chronic toxicity, and the promotion of antimicrobial resistance. Conventional wastewater treatment plants (WWTPs) remove 70–90% of ACT but less than 30% of DCF, primarily because these systems were not designed to target low-concentration, recalcitrant micropollutants. As a result, pharmaceuticals frequently pass into treated effluents, highlighting the need for advanced, sustainable, and passive treatment solutions. Permeable reactive barriers (PRBs) have emerged as a promising technology for the interception and removal of pharmaceuticals from both wastewater treatment plant effluents and groundwater. This review provides a comprehensive assessment of ACT and DCF occurrence, environmental behavior, and ecotoxicological risks, followed by a detailed evaluation of PRB performance using advanced reactive media such as geopolymers, activated carbon, carbon nanotubes, and hybrid composites. Reported removal efficiencies exceed 90% for ACT and 70–95% for DCF, depending on media composition and operating conditions. The primary removal mechanisms include adsorption, ion exchange, π–π interactions, hydrogen bonding, and redox transformation. The novelty of this review lies in systematically synthesizing recent laboratory and pilot-scale findings on PRBs for pharmaceutical removal, identifying critical knowledge gaps including long-term field validation, media regeneration, and performance under realistic wastewater matrices and outlining future research directions for scaling PRBs toward full-scale implementation. The study demonstrates that PRBs represent a viable and sustainable tertiary treatment option for reducing pharmaceutical loads in aquatic environments.
- Evaluating soil phosphorus tests and nutrient limitations in Mediterranean pasturesPublication . Arrobas, Margarida; Raimundo, Soraia; Sawimbo, Almeida; Rodrigues, Nuno; Jongen, Marjan; Teixeira, Ricardo F.M.; Domingos, Tiago; Rodrigues, Manuel ÂngeloReliable phosphorus (P) fertilization guidelines for Mediterranean pastures remain uncertain due to inconsistent soil testing methods, the complexity of mixed-species systems, and the neglect of other limiting nutrients. This study primarily aimed to identify reliable soil P tests and secondarily to explore potential nutrient limitations by assessing seven acidic Portuguese soils under Mediterranean conditions using subterranean clover (Trifolium subterraneum) and ryegrass (Lolium multiflorum) as model species. Five P extraction methods (Egner-Riehm, Olsen, Bray II, Mehlich I, and anion exchange resin) were compared alongside a pot experiment with clover, ryegrass, and their mixture. Biomass production, plant nutritional status, and soil-plant relationships were used to assess nutrient availability and plant response. The Egner-Riehm method proved the most reliable, showing linear correlation with the resin method (R2 = 0.89), widely regarded as the best indicator of soil P bioavailability. P availability was therefore not a limiting factor for biomass production. The lowest soil P content (54 mg kg-1 P2O5, Egner-Riehm) was close to the medium threshold (50-100 mg kg-1), yet plant growth indicated adequacy. In contrast, nitrogen (N) emerged as the main growth constraint, and boron (B) deficiency further restricted clover performance, confirming additional nutritional imbalances. A linear relationship between soil and tissue B supported its association with DMY, and tissue B levels were below sufficiency thresholds. Overall, in acidic Portuguese soils under Mediterranean conditions, soil P was adequate, whereas N and B acted as key constraints to productivity. These findings support more accurate and efficient fertilization strategies for Mediterranean pastures.
- Bioactive metabolites from algae: occurrence, extraction techniques, functional properties, food applications and therapeutic prospectsPublication . Pereira, Cláudia S.G.P.; Carpena, M.; Barreira, João C.M.; Silva, Cristiana F.C.; Prieto Lage, Miguel A.; Oliveira, BeatrizThis review critically summarizes the major algal-derived bioactive compounds, including phenolics compounds (e.g., phlorotannins, bromophenols, phenolic acids), sterols (e.g., fucosterol, beta-sitosterol), and polysaccharides (e. g., carrageenan, fucoidan, ulvan), as well as key pigments such as astaxanthin, beta-carotene, lutein, violaxanthin, zeaxanthin, and chlorophylls. In addition, state-of-the-art extraction technologies including supercritical fluid extraction, pressurized liquid extraction, ultrasound assisted, microwave assisted, surfactant assisted, and enzyme assisted extractions, and carbon dioxide-expanded liquids) were systematically compared considering key parameters such as yield, versatility, compound selectivity, extract quality, cost-effectiveness, technical complexity, operational conditions, and sustainability principles. Based on this comparative evaluation, ultrasound assisted extraction emerged as the most versatile among advanced technologies, same as enzyme assisted extraction within ulterior techniques. The potential incorporation of these bioactive compounds into novel algaebased food systems, either as functional ingredients or texturizing agents, was also discussed. Moreover, their relevance in alternative therapeutic applications was also highlighted, with a proof-of-concept focused on the antiviral potential of sulfated polysaccharides (carrageenan, fucoidan, and ulvan). Overall, algae-based ingredients were found to be promising biofunctional agents for innovative food products, nutraceutical formulations, and therapeutic alternatives.
- Integrated experimental, process simulation, and techno-economic assessment of biogas upgrading via pressure/vacuum swing adsorptionPublication . Karimi, Mohsen; Siqueira, Rafael M.; Shirzad, Mohammad; Ferreira, Alexandre F.P.; Rodrigues, Alírio; Silva, José A.C.This study presents an integrated approach in biogas upgrading technology through the development and optimization of a shaped MIL-160(Al)-based pressure/vacuum swing adsorption (PSA/VPSA) system. Combining detailed experimental investigations with comprehensive process modeling and techno-economic analysis, we demonstrate a complete pathway from fundamental dynamic adsorption to industrial implementation. Breakthrough tests reveal notable CO2/CH2 separation performance with shaped MIL-160(Al), while 23 cyclic PSA experiments achieved over 90 % methane purity. Advanced process modeling, validated with less than 5 % deviation from experimental data, enables successful scale-up to industrial VPSA configurations, where 38 distinct cases were evaluated to identify an optimal system producing 99.81 % pipeline-quality biomethane with 92.6 % recovery. Our holistic techno-economic assessment reveals the system's acceptable economic viability, with the total capital expenditure (CapEx) of $14.33 million. Accordingly, this work provides clear methodological insights that strengthen the understanding of MIL-160(Al)-based PSA/VPSA process and support its potential application for biogas upgrading.
- A Spectrochemically Driven Study: Identifying Phenolic-Rich Extracts from Helichrysum stoechas, Lavandula pedunculata, and Thymus mastichina with Potential to Revert Skin Aging EffectsPublication . Marques, Mário Pedro; Landim, Euclides; Varela, Carla; Costa, Ricardo M. F. da; Marques, Joana; Carvalho, Luís A. E. Batista de; Silva, Ana; Cruz, Maria Teresa; André, Rebeca; Rijo, Patrícia; Dias, Maria Inês; Carvalho, Aida; Oliveira, Paulo J.; Cabral, CéliaPlants inhabiting mediterranean-influenced climatic zones, like Helichrysum stoechas (L.) Moench subsp. stoechas, Lavandula pedunculata (Mill.) Cav., and Thymus mastichina (L.) L. subsp. mastichina, have been scarcely investigated regarding their richness in phenolic compounds, herein explored as sources of skin anti-aging compounds. In this investigation, Fourier transform infrared spectroscopy (FTIR) in attenuated total reflectance (ATR) mode and high-performance liquid chromatography coupled with diode-array detection and electrospray ionization tandem mass spectrometry (HPLC-DAD-ESI/MSn) were employed to chemically characterize the hydroethanolic extracts (HEs), and their cell-free antioxidant potential was screened. Thereafter, non-toxic concentrations of HEs were determined in human skin cells using Alamar blue® and Sulforhodamine B assays. The cytoprotective and antioxidant effects of HEs were assessed in tert-butyl hydroperoxide-stimulated fibroblasts, their anti-inflammatory potential was studied in lipopolysaccharide-injured macrophages, and enzymatic inhibition assays were performed. Notably, the irritant effects of HEs were tested according to Test Guideline No. 439 of the Organization for Economic Co-operation and Development (OECD). The major compounds identified in the T. mastichina and L. pedunculata HEs were rosmarinic and salvianolic acid derivatives, while H. stoechas HE was mainly composed of caffeoyl and feruloyl derivatives, and O-glycosylated flavonoids. T. mastichina (≤0.4 mg/mL) exhibited significant cytoprotective, anti-inflammatory, and antioxidant effects, as well as remarkable anti-hyaluronidase activity. Shedding light on the quantitative and qualitative chemical picture of these HEs highlighted T. mastichina as a promising candidate to target skin aging effects, which correlates with its phenolic content. Further investigation is warranted regarding its anti-aging pharmacological activity, which could lead to the development of plant-based skin anti-aging products.
- Insect-based biopolymers from Orthoptera: An emerging resource for sustainable and active food packagingPublication . Zannou, Oscar; Conti, Barbara; Abenaim, Linda; Lopes, João Krauspenhar; Barciela, Paula; Roriz, Custódio Lobo; Prieto Lage, Miguel A.; Heleno, Sandrina A.; Carocho, Márcio; Goksen, Gulden; Lopes, João KrauspenharThe challenges of the current era in food packaging have led to a continuous diversification of sources of biopolymers. Edible Orthopterans have garnered significant interest worldwide as potential sources of biopolymers for the production of biodegradable and active packaging. These insect species have emerged as a potential and sustainable substitute for conventional protein and chitosan sources due to their high nutritional content and eco-friendly cultivation methods. Scope and approach: This review explores the recent development of sustainable sources of biopolymers, focusing on the edible Orthopterans. We elucidated the main edible Orthoptera species used as the sources of biopolymers for food packaging, extraction methods of biopolymers from Orthopterans, fabrication of packaging materials and their characteristics. Moreover, we analyzed the current applications of these packaging materials for food preservation. Furthermore, we proposed a critical analysis of the health issues, consumer acceptance, legislation and commercialization, along with the future directions. Key findings and conclusions: Acheta domesticus, Gryllodes sigillatus and Locusta migratoria are the Orthopterans used as sources of biopolymers for the production of packaging materials. Chitosan and proteins are the major Orthopteran-derived biopolymers used for the production of sustainable, edible and bioactive food packaging materials. Emerging technologies have shown potential to enhance extraction efficiency while preserving the functionality of the biopolymers. These biopolymers exhibit remarkable potential for the development of food packaging materials, due to their biodegradability, biocompatibility, and functional properties. More research initiatives and awareness are needed to facilitate the implementation of Orthopteran-derived biopolymers as active packaging materials.
- Interpreting Machine Learning Models with SHAP Values: Application to Crude Protein Prediction in Tamani Grass PasturesPublication . Monteiro, Gabriela Oliveira de Aquino; Difante, Gelson dos Santos; Montagner, Denise Baptaglin; Euclides, Valéria Pacheco Batista; Castro, Marina; Rodrigues, Jéssica Gomes; Pereira, Marislayne de Gusmão; Ítavo, Luís Carlos Vinhas; Campos, Jecelen Adriane; Costa, Anderson Bessa da; Matsubara, Edson TakashiMachine learning models such as XGBoost show strong potential for predicting pasture quality metrics like crude protein (CP) content in tamani grass (Panicum maximum). However, their 'black box' nature hinders practical adoption. To address this limitation, this study applied SHapley Additive exPlanations (SHAP) to interpret an XGBoost model and uncover how management practices (grazing interval, nitrogen fertilization, and pre- and post-grazing heights) and environmental factors (precipitation, temperature, and solar radiation) jointly influence CP predictions. Data were divided into 80% for training/validation and 20% for testing. Model performance was assessed with stratified 5-fold cross-validation, and hyperparameters were tuned via grid search. The XGBoost model yielded a Pearson correlation coefficient (r) of 0.78, a mean absolute error (MAE) of 1.45, and a coefficient of determination (R2) of 0.57. The results showed that precipitation in the range of 100-180 mm increased the predicted CP content. Application of 240 kg N ha-1 year-1 positively affected predicted CP, whereas a lower dose of 80 kg N ha-1 year-1 had a negative impact, reducing predicted levels of CP. These findings highlight the importance of integrated management strategies that combine grazing height, nitrogen fertilization, and grazing intervals to optimize crude protein levels in tamani grass pastures.
- Comprehensive assessment of PM10 from home heating using different appliances and biomass fuels: Chemical composition, oxidative potential, and ecotoxicityPublication . Cipoli, Yago Alonso; Vicente, Estela D.; Charres, Isabella; Evtyugina, Margarita; Alfosea-Simón, Marina; Lucarelli, Franco; Kováts, Nora; Ryšavý, Jiří; Feliciano, Manuel; Alves, CéliaThe European Union has implemented policies to promote renewable energy, with an emphasis on biomass for heat generation. However, residential biomass combustion is a major source of particulate matter (PM10), and its chemical constituents pose health concerns worldwide. This study characterised the organic and inorganic composition, oxidative potential (OP), and ecotoxicity of PM10 indoors and outdoors during the operation of two heating appliances fuelled with different types of biomasses: a modern hydronic stove and a traditional wood stove. PM10 concentrations were higher in the room equipped with the traditional system during the combustion of briquettes (95.9 f 74.9 mu g m-3) and firewood (50.1 f 25.6 mu g m- 3), compared to the modern stove using pellets (27.1 f 11.8 mu g m-3) and olive stone (23.0 f 4.5 mu g m- 3). While element oxides accounted for similar PM10 mass fractions (7 %), the wood stove produced higher levels of organic constituents, including carbonaceous fractions, polycyclic aromatic hydrocarbons (PAHs), quinones and saccharides. Significant correlations between OP assays and concentrations of PAHs, quinones and phenolic compounds were found in the room equipped with wood stove, with indoor to outdoor (I/O) ratios higher than 1. In contrast, I/O ratios below 1 were observed for OP in PM10 samples collected during hydronic stove operation. Ecotoxicity assays using Aliivibrio fischeri classified indoor PM10 from the wood stove as 'very toxic', whereas samples from the hydronic stove exhibited lower toxicity and OP levels. These findings highlight the need to reconcile renewable energy goals with air quality and public health.