Publications

Abstract Active and abandoned mining sites are significant sources of heavy metals and metalloid pollution, leading to serious environmental issues. This study assessed the environmental risks posed by potentially toxic elements (PTEs), specifically arsenic (As) and antimony (Sb), in the Technosols (mining residues) of the former Pej & atilde;o coal mine complex in Northern Portugal, a site impacted by forest wildfires in October 2017 that triggered underground combustion within the waste heaps. Our methodology involved determining the pseudo-total concentrations of As and Sb in the collected heap samples using microwave digestion with aqua regia (ISO 12914), followed by analysis using hydride generation-atomic absorption spectroscopy (HG-AAS). The concentrations of As an Sb ranging from 31.0 to 68.6 mg kg-1 and 4.8 to 8.3 mg kg-1, respectively, were found to be above the European background values reported in project FOREGS (11.6 mg kg-1 for As and 1.04 mg kg-1 for Sb) and Portuguese Environment Agency (APA) reference values for agricultural soils (11 mg kg-1 for As and 7.5 mg kg-1 for Sb), indicating significant enrichment of these PTEs. Based on average Igeo values, As contamination overall was classified as unpolluted to moderately polluted while Sb contamination was classified as moderately polluted in the waste pile samples and unpolluted to moderately polluted in the downhill soil samples. However, total PTE content alone is insufficient for a comprehensive environmental risk assessment. Therefore, further studies on As and Sb fractionation and speciation were conducted using the Shiowatana sequential extraction procedure (SEP). The results showed that As and Sb levels in the more mobile fractions were not significant. This suggests that the enrichment in the burned (BCW) and unburned (UCW) coal waste areas of the mine is likely due to the stockpiling of lithic fragments, primarily coals hosting arsenian pyrites and stibnite which largely traps these elements within its crystalline structure. The observed enrichment in downhill soils (DS) is attributed to mechanical weathering, rock fragment erosion, and transport processes. Given the strong association of these elements with solid phases, the risk of leaching into surface waters and aquifers is considered low. This work underscores the importance of a holistic approach to environmental risk assessment at former mining sites, contributing to the development of sustainable remediation strategies for long-term environmental protection.

Abstract (1) Background: The under-representation of women in STEM fields, particularly in areas such as physics and computing, is far from being resolved. This gender gap has complex causes. This research was carried out to understanding how physics teachers in Portugal are aware of the existence of a gender gap and how they use or are willing to use girl-friendly strategies in their lessons; (2) Methods: A sample of 55 Portuguese physics and chemistry teachers from the third cycle of basic education and secondary education and an 8-item survey were used. (3) Results: The results show that most teachers perceive girls' participation in physics as satisfactory, and that girls do not perceive a gender gap and are interested in the subject, but may not be aware of the concept and application of girl-friendly strategies. (4) Conclusions: No correlation was found between gender or years of service and interest in the topic of girl-friendly strategies. Further research with a more diverse sample is needed to generalize these findings.

Abstract S-1 MitoPlates™ from Biolog enable the characterization of mitochondria’s function in live cells by measuring the rates of electron flow into and through the electron transport chain from different NADH or FADH2 producing metabolic substrates. This technology uses 96-well microplates pre-coated with triplicate repeats of a set of 31 substrates. Those 31 metabolic substrates have different routes of entry into the mitochondria, use different transporters, and are also oxidated by different dehydrogenases, producing reducing equivalents in the form of NADH or FADH2. The electrons produced upon oxidation of NADH or FADH2 at complex I or II, respectively, then travel to cytochrome c, where a tetrazolium redox dye (MC) can act as terminal acceptor, turning purple and absorbing at 590 nm. This mechanism allows the evaluation of cellular substrate preference by following the kinetics of MC reduction in the presence of selected substrates. In this chapter, we describe the step-by-step protocol to prepare an experiment using MitoPlate S-1 array and the OmniLog instrument to assess the metabolism of human dermal fibroblasts. We also give detailed information on how to analyze the raw data generated by the Biolog Data Analysis software to extract meaningful information and produce useful data visualizations, using reproducible methods based on a single structured dataset. © The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature 2025.

Abstract Metabolic syndrome (MetS) has been associated with disruptions in tissue mechanical homeostasis and inflammatory and metabolic derangements. However, the direct correlation between metabolic alterations and changes in tissue stiffness, and whether they could play a role as upstream initiators of disease pathology remains to be investigated. This emerging concept has yet to be put into clinical practice as many questions concerning the interplay between extracellular matrix mechanical properties and regulation of metabolic pathways remain unsolved. This review will highlight key foundational studies examining mutual regulation of cell metabolism and mechanotransduction, and opening questions lying ahead for better understanding MetS pathophysiology.

Abstract Mitochondrial dysfunction and increased reactive oxygen species (ROS) generation play an import role in different human pathologies. In this context, mitochondrial targeting of potentially protective antioxidants by their coupling to the lipophilic triphenylphosphonium cation (TPP) is widely applied. Employing a six-carbon (C6) linker, we recently demonstrated that mitochondria-targeted phenolic antioxidants derived from gallic acid (AntiOxBEN2) and caffeic acid (AntiOxCIN4) counterbalance oxidative stress in primary human skin fibroblasts by activating ROS-protective mechanisms. Here we demonstrate that C6-TPP (but not AntiOxBEN2 and AntiOxCIN4) induce cell death in human skin fibroblasts. This indicates that C6-TPP cytoxocity is counterbalanced by the antioxidant moieties of AntiOxBEN2 and AntiOxCIN4. Remarkably, C6-TPP and AntiOxBEN2 (but not AntiOxCIN4) induced a glycolytic switch, as exemplified by a reduced cellular oxygen consumption rate (OCR), increased extracellular acidification rate (ECAR), elevated extracellular lactate levels, and higher protein levels of glucose transporter 1 (GLUT-1). This switch involved activation of AMP-activated protein kinase (AMPK) and fully compensated for the loss in mitochondrial ATP production by sustaining cellular ATP content. When glycolytic switch induction was prevented ( i.e. by using a glucose-free, galactose-containing medium), AntiOxBEN2 induced cell death whereas AntiOxCIN4 did not. We conclude that, despite their similar chemical structure and antioxidant capacity, AntiOxBEN2 and AntiOxCIN4 display both common (redox-adaptive) and specific (bioenergetic-adaptive) effects.

Abstract In the last decades, major changes in ecosystems related to industrial development and environmental modifications have had a direct impact on mammalian fertility, as well as on biodiversity. It is widely demonstrated that all these changes impair reproductive function. Several studies have connected the increase of reactive oxygen species (ROS) generated in mitochondria to the recently identified decline of fertility due to various factors, including heat stress. The study of antioxidants, and especially of mitochondria targeted antioxidants, has been focused on identifying more efficient and less toxic therapies that could circumvent fertility problems. These antioxidants can be derived from natural compounds in the diet and delivered to the mitochondria in more effective forms, providing a much more natural therapy. The use of mitochondriotropic diet-based antioxidants in assisted reproductive technologies (ART) may be an important way to overcome low fertility, allowing the conservation of animal biodiversity and productivity. This paper provides a concise review of the current state of the art on this topic, with a particular focus on the antioxidants mitoquinone, AntiOxBEN2, AntiOxCIN4, urolithin A and piperine, and their effects on bovine and other animal species.

Abstract BackgroundExtracellular matrix (ECM) stiffness is increasingly recognized as a critical regulator of cellular behaviour, governing processes such as proliferation, differentiation, and metabolism. Neurodegenerative diseases are characterized by mitochondrial dysfunction, oxidative stress, impaired autophagy, and progressive softening of the brain tissue, yet research into how mechanical cues influence cellular metabolism in this context remains scarce.Materials and MethodsIn this study, we evaluated the long-term effects of brain-compliant, soft ECM on mitochondrial bioenergetics, redox balance, and autophagic capacity in human neuroblastoma (SH-SY5Y) and mouse hippocampal (HT22) cell lines, as well as primary mouse neurons.ResultsWe observed that prolonged exposure to soft ECM does not impact cell proliferative capacity of neuronal cells but results in mitochondrial bioenergetic dysfunction, redox imbalance, and disrupted autophagic flux. These findings were consistently validated across both human and mouse neuronal cells. Our data indicate a decreased maximal autophagic capacity in cells exposed to long-term soft ECM, potentially due to an imbalance in autophagosome formation and degradation, as demonstrated by decreased LC3 II levels following chloroquine-induced autophagic flux inhibition. This impairment in autophagy was coupled with increased cellular oxidative stress, further indicating metabolic alterations.ConclusionsThese findings emphasize the critical role of ECM stiffness in regulating neuronal cell metabolism and suggest that prolonged exposure to soft ECM may mimic key aspects of neurodegenerative disease pathology, thereby enhancing the physiological relevance of in vitro models. This study underscores the necessity for further research into ECM mechanics as a contributing factor in neurodegenerative disease progression and as a potential target for therapeutic strategies.

Abstract Integrated science, technology, engineering, and mathematics (STEM) education is essential for preparing students to address real-world challenges through interdisciplinary thinking and problem-solving. However, preservice teachers (PSTs) often face challenges in integrating STEM disciplines, particularly engineering, technology, and mathematics. This study explores how Arduino-based problem-solving activities enhance interdisciplinary STEM integration in PST training, focusing on the epistemological, psychological, and didactic dimensions of STEM education. Conducted within a chemistry teacher training course, the study involved eight PSTs in theoretical and practical activities, including designing experiments on ocean acidification. The findings reveal that Arduino-based activities effectively foster interdisciplinary connections by integrating chemistry with technology and engineering while bridging theoretical knowledge and practical applications. Persistent challenges, such as limited mathematics integration and familiarity with engineering concepts, highlight the need for targeted interventions in teacher training. This study underscores the potential of problem-solving and technology-enhanced approaches to equip teachers with the skills to design and implement interdisciplinary STEM lessons, preparing them for the demands of 21st-century education.

Abstract Groundwater is a critical resource in semi-arid regions like Morocco's Guire Basin, yet pollution and overexploitation threaten its sustainability. This study evaluates the groundwater quality of the Guire aquifer (Eastern High Atlas) using an integrated approach combining hydrochemical, isotopic (delta O-18, delta H-2, delta C-13), multivariate statistical, and Geographic Information System (GIS) analyses alongside the Water Quality Index (WQI). Sixteen wells were monitored for physicochemical parameters (pH: 7-7.9; EC: 480-3004 mu S/cm; BOD5: 1.03-30.5 mg/L; COD: 10.2-45.75 mg/L) and major ions, revealing widespread exceedances of Moroccan standards for Cl-, HCO3-, Mg2+, Ca2+, and NH4+. WQI classified 81% of samples as Poor to Unsuitable for drinking (WQI: 51-537), driven by elevated Cl-, Na+, and SO42- from Triassic evaporite dissolution and NO3- (up to 45 mg/L) from agricultural runoff. Stable isotopes (delta O-18: -7.73 parts per thousand to -5.08 parts per thousand; delta H-2: -66.14 parts per thousand to -44.20 parts per thousand) indicate Atlantic-influenced recharge at 900-2200 m altitudes, with a delta O-18-delta H-2 slope of 5.93 reflecting evaporation during infiltration. Strontium (Sr2+/Ca2+: 0.0024-0.0236) and bromide (Br/Cl: 8.47 x 10(-5)-9.88 x 10(-4)) ratios further confirm evaporitic dominance over anthropogenic contamination. This work provides actionable insights for policymakers, advocating for targeted restrictions on fertilizers, enhanced monitoring near evaporite zones, and artificial recharge initiatives. By linking geogenic/anthropogenic contamination to governance strategies, this study advances sustainable groundwater management in semi-arid regions.

Abstract <jats:p>Mozambique soils are known for having an unbalanced agronomic and environmental composition that results in poor agricultural production yields. However, agriculture is the main economic activity of Mozambique, and soils must be characterised for their elemental deficiencies and/or excesses. This paper sampled nine farms from the Manica and Sussundenga districts (Manica province) in three campaigns in 2021/2022, 2022/2023, and 2023/2024 (before and after the rainy seasons). They were subjected to a physical–chemical analysis to assess their quality from the fertility and environmental contamination point of view. Attending to the physical–chemical properties analysed, and for all the soils and sampling campaigns, a low concentration below the limit of detection for B of &lt;0.2 mg/Kg for the majority of soils and a low concentration of Al &lt; 0.025 mg/Kg for all the soils were obtained. Also, higher concentrations for the majority of soils for the Ca between 270 and 1634 mg/Kg, for the Mg between 41 and 601 mg/Kg, for the K between 17 and 406 mg/Kg, for the Mn between 13.6 and 522 mg/Kg, for the Fe between 66.3 and 243 mg/Kg, and for the P between &lt;20 and 132 mg/Kg were estimated. In terms of texture and for the sand, a high percentage between 6.1 and 79% was found. In terms of metal concentrations and for all the soils of the Sussundenga district and sampling campaigns, a concentration above the reference value concentration for the Cr (76–1400 mg/Kg) and a concentration below the reference value concentration for the Pb (5–19 mg/Kg), Ba (13–120 mg/Kg) and for the Zn (10–61 mg/Kg) were evaluated. A multivariate data analysis methodology was used based on cluster and discriminant analysis. The analysis of twenty-three physical–chemical variables of the soils suggested four clusters of soils characterised by deficiencies and excess elements that must be corrected to improve the yield and quality of agricultural production. Moreover, the multivariate analysis of the metal composition of soil samples from the second and third campaigns, before and after the rainy season, suggested five clusters with a pristine composition and different metal pollutant compositions and concentrations. The information obtained in this study allows for the scientific comprehension of agricultural soil quality, which is crucial for designing agronomic and environmental corrective measures to improve food quality and quantity in the Manica and Sussundenga districts and ensure environmental, social, and economic sustainability.</jats:p>