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- Variation in reporting of heatstroke mortality: evidence from a multi-country studyPublication . Tobias, Aurelio; Honda, Yasushi; Madaniyazi, Lina; Alhamad, Barrak; Lavigne, Erik; Roye, Dominic; Tong, Shilu; de Sousa Zanotti Stagliorio Coelho, Micheline; Huber, Veronica; Urban, Ales; das Neves Pereira da Silva, Susana; Achilleos, Souzana; Parks, Robbie M.; Iñiguez, Carmen; Masselot, Pierre; Vicedo-Cabrera, Ana M.; Armstrong, Ben; Gasparrini, Antonio; Hashizume, Masahiro; Multi-City Multi-Country Collaborative Research NetworkBackground: Heatstroke represents the most severe manifestation of heat exposure. Heatstroke is rare and under-reported, resulting in limited empirical data on its global incidence and burden. This study aimed to examine geographical variations and temporal trends in reported heatstroke mortality across multiple countries. Methods: We collected annual heatstroke mortality data from 34 countries participating in the Multi-Country Multi-City Collaborative Research Network between 2000 and 2022, using the ICD-10 code X30. Country-specific mortality rates were estimated using Poisson regression, alongside analyses of annual trends and associations with mean warm-season temperature. We also assessed the proportion of heatstroke deaths relative to both overall heat and extreme heat-attributable all-cause mortality. Findings: Heatstroke mortality rates varied widely across countries, with Japan reporting the highest rate (5·81 per 1 million population; 95% CI 4·43-7·62), followed by Cyprus (2·51; 1·36-4·61), and China (2·42; 1·21-4·85). By contrast, most countries in Europe, South America, and southeast Asia reported rates of less than one death per 1 million population. Heatstroke mortality increased over time in several countries and was associated with warm-season temperatures in most regions. The proportion of heatstroke deaths relative to overall heat-attributable mortality ranged from less than 1% in many countries to as close to 24% in Japan. When analyses were restricted to deaths attributable to extreme heat, the proportion of heatstroke deaths increased substantially. Interpretation: Our broad international assessment of heatstroke mortality highlights its distinct patterns compared with overall heat-attributable mortality. The observed variability likely reflects differences in recognition, reporting, and diagnostic practices, while climate exposure and health system capacity influence whether heat-related deaths are identified and recorded as heatstroke.
- Evidence of Endocrine Disruption and Oxidative Stress in Mytilus galloprovincialis Exposed to 17α-EthinylestradiolPublication . Copeto, Sandra; Ferreira, Inês João; Mansilha, Catarina; Melo, Armindo; Motta, Carla; Silva, Marco; Diniz, MárioThe presence of endocrine-disrupting compounds (EDCs) in aquatic environments has raised significant concerns, particularly regarding their impact on marine biota. Among these compounds, 17α-ethinylestradiol (EE2), a synthetic estrogen widely used in oral contraceptives, is highly persistent and biologically active at very low concentrations. This study evaluated the effects of EE2 exposure on oxidative stress responses and endocrine disruption in Mytilus galloprovincialis, exposed for 28 days to three EE2 concentrations (10, 30, and 300 ng·L−1). Biomarkers of oxidative stress, including the enzymatic activities of superoxide dismutase (SOD), catalase (CAT), and glutathione-S-transferase (GST), as well as Lipid Peroxidation (MDA levels), total ubiquitin (UBI) and the endocrine disruption marker, vitellogenin-like protein (VTG) were assessed. Results showed significant increase in GST and a decrease in CAT activities followed by an elevation at 300 ng·L−1, slightly higher than control values. Overall, results suggest an enhanced oxidative challenge. No significant changes were detected in MDA and UBI levels. VTG-like protein levels increased according to the EE2 concentrations tested, suggesting an effect on the mussel’s endocrine system. These results show the activation of detoxification and antioxidant defense mechanisms in exposed mussels as a response to mitigate oxidative stress damage. Furthermore, it highlights the importance of using biomarkers in pollution monitoring studies and environmental risk assessment.
- Association of mortality and combined oxidative capacity of ozone and nitrogen dioxidePublication . Niu, Yue; Chen, Renjie; Sera, Francesco; Urban, Aleš; Vicedo-Cabrera, Ana Maria; Honda, Yasushi; Huber, Veronika; Guo, Yuming; Tong, Shilu; Coelho, Micheline de Sousa Zanotti Staglior; Saldiva, Paulo Hilario Nascimento; Lavigne, Eric; Correa, Patricia Matus; Ortega, Nicolás Valdés; Osorio, Samuel; Achilleos, Souzana; Roye, Dominic; Jaakkola, Jouni J. K.; Ryti, Niilo; Pascal, Mathilde; Schneider, Alexandra; Breitner-Busch, Susanne; Entezari, Alireza; Mayvaneh, Fatemeh; Raz, Raanan; Sheng Ng, Chris Fook; Hashizume, Masahiro; Carrasco, Gabriel; das Neves Pereira da Silva, Susana; Madureira, Joana; Holobaca, Iulian-Horia; Kim, Ho; Lee, Whanhee; Tobias, Aurelio; Íñiguez, Carmen; Guo, Yue Leon; Pan, Shih-Chun; Li, Shanshan; Masselot, Pierre; Bell, Michelle L.; Zanobetti, Antonella; Schwartz, Joel; Gasparrini, Antonio; Kan, HaidongOzone (O3) and nitrogen dioxide (NO2) are two common gaseous pollutants that both possess oxidizing properties with consequences for human health and have an inextricable chemical relationship that could have distinct public health impacts when considered in combination. We examined the short-term associations of the combined oxidative capacity of O3 and NO2 (represented by Oxwt, the average of O3 and NO2 concentrations weighted by their standard electrode potential) with total, cardiovascular and respiratory mortality in 380 cities across 23 countries or regions between 1985 and 2020. Over 2 days (LAG01), a 10-ppb increase in Oxwt concentration was associated with an increase of 0.82% (95% confidence interval (CI): 0.55%, 1.10%) in total mortality, 1.09% (95% CI: 0.83%, 1.35%) in cardiovascular mortality and 0.88% (95% CI: 0.31%, 1.45%) in respiratory mortality. We also observed variations in this association by geographic region and study period. More deaths were attributable to Oxwt than to either O3 or NO2 but fewer than the sum of the two. Thus, Oxwt might be a valuable indicator for use in public health efforts to capture the combined effects of O3 and NO2.
- Assessing global factors associated with tropical cyclone-related mortality: A population-based longitudinal studyPublication . Huang, Wenzhong; Yang, Zhengyu; Otto, Christian; Mengel, Matthias; Hales, Simon; Zhang, Yiwen; Xu, Rongbin; Bell, Michelle L.; Gasparrini, Antonio; Kan, Haidong; Sera, Francesco; Schwartz, Joel; Lavigne, Eric; Hundessa, Samuel; Yu, Wenhua; Carlos Chua, Paul Lester; Seposo, Xerxes; Goodman, Patrick; Zeka, Ariana; Hashizume, Masahiro; Z S Coelho, Micheline S.; Xu, Zhihu; Ye, Tingting; Yu, Pei; Wu, Yao; Wen, Bo; Liu, Yanming; Li, Shanshan; Guo, Yuming; MCC CollaboratorsBackground: The underlying factors associated with the substantial global tropical cyclone (TC)-related mortality burden, characterized by its highly variable spatiotemporal patterns, remain unclear. We aimed to identify and assess the key factors associated with TC-related mortality on a global scale. Methods: We collected mortality records from 2034 locations in 68 countries/territories across five continents (2000-2019) to identify and assess the key factors associated with TC-related mortality on a global scale. Bayesian ensemble models were applied to estimate the associated mortality for each TC event in each location. A random forest regression (RFR) model was employed to assess the relative statistical importance of TC and location characteristics, as well as their interactions, regarding the TC-related mortality. Findings: For TC physical characteristics, the TC-associated cumulative rainfall consistently exhibited stronger influence on mortality than TC-induced surge-driven flood depth or maximum sustained windspeed. However, sociodemographic factors, including population traits (e.g., population density, proportion of the population aged ≤ 9 years, percent of the population aged ≥ 65 years) and socioeconomic and infrastructure development (e.g., built-up ratio), showed greater relative importance than the TC physical attributes and accounted for the majority of the explained variations in TC-related mortality. Furthermore, cumulative rainfall interacted strongly with local sociodemographic conditions and was potentially the most important associated factor for disparities in mortality arising from factor interactions. Interpretation: The findings highlight the critical role of sociodemographic factors in explaining the global spatiotemporal variability of TC-related mortality, surpassing the relative importance of TC intensity. TC-related rainfall could be a key associated physical factor of the global mortality burden.
- Sand Quality on Portuguese Blue Flagged Beaches: Fungal and Faecal Contamination Across Two Bathing SeasonsPublication . Silva, Ana Margarida; Sarioglou, Konstantina; das Neves Pereira da Silva, Susana; Viegas, Carla; Ribeiro, Edna; Rebelo, Maria Teresa; Brandão, JoãoThere is growing concern about the quality of sand on beaches, as users tend to spend most of their time on the sand rather than in the water. Numerous pathogenic agents have reportedly been isolated from sand, including bacteria, nematodes and opportunistic fungi. The ability of sand to retain pollutants and facilitate the transmission of pathogens raises public health concerns. We analysed sand-monitoring data from the 2024 and 2025 bathing seasons on Blue Flag beaches to find trends and patterns in total fungal counts, enterococci, and E. coli. The values recorded for microorganisms showed considerable variability, which may reflect the possible combined influence of multiple climatic, environmental, and anthropogenic factors contributing to their presence in beach sand. Our findings suggest that the total fungal count on coastal beaches may be influenced by periods of rainfall, which increases the fungal load in the sand. Values recorded from inland beaches vary considerably between beaches which may reflect the influence of local environmental characteristics, particularly vegetation and beach morphology, although the smaller number of inland samples also makes it difficult to define clear patterns and consistent reference values for this parameter. Bacterial indicators may be particularly influenced by occasional anthropogenic disturbances and contamination events. This study adds significantly to the understanding of the microbiological quality of beach sand, encouraging the integration of sand monitoring into environmental surveillance and management programmes.
- Predicted no effect concentrations of antifungals for wastewater management and agricultural usePublication . Gil, D.; José, S.; Ascenso, A.; Babič, M. Novak; Segal, E.; Meletiadis, J.; Gangneux, J.P.; Weiskerger, C.J.; Solo-Gabriele, H.M.; Valério, E.; Brandão, J.Antifungal resistance is an on-growing public health concern due to the difficulty in managing or treating medical conditions that often favour fatal fungal infections. The changing climate and globalisation, which increase fungal persistence and propagation, adds to that concern. Wastewater disposal is one potential source to the environment as antifungals are released into it. Considering that most fungal infections originate from the environment and considering the One Health principle, introducing antifungals through wastewater effluents has the potential to promote the emergence and dissemination of antifungal resistance. The objective of this study was to generate knowledge that can assist regulating the release of antifungals in the environment by quantifying predicted no-effect concentrations (PNECs) that would not promote antifungal resistance. For this purpose, a systematic review was performed to consolidate information on antifungals released to the environment and respective concentrations. The systematic literature review followed Preferred Reporting Items for Systematic literature reviews and Meta Analyses extension for Scoping Reviews (PRISMA-SLR). The analysis of 122 reviewed articles using this approach showed high concentrations and dispersion of antifungals in water, wastewater or soil. This highlights their potential dispersion in the environment, thus increasing the potential of fungal antimicrobial resistance. Due to the lack of PNEC values using fungi as model organisms in this review, PNECs for 17 antifungals were calculated using as model, as it is done for clinical purposes. We consider that the antifungal PNECs calculated and consolidated from the literature can be used to prioritise them for regulation and to determine acceptable levels in wastewater effluents.
- In vitro and in vivo assessment of nanoceria biocompatibility for their safe use in nervous system applicationsPublication . Fernández-Bertólez, Natalia; Martínez, Luisa; Ramos-Pan, Lucía; Touzani, Assia; Costa, Carla; Laffon, Blanca; Valdiglesias, VanessaNanoceria, or cerium dioxide nanoparticles (CeO NP), are increasingly employed in a number of industrial and commercial applications. Hence, the environmental presence of these nanoparticles is growing progressively, enhancing the global concern on their potential health effects. Recent studies suggest that nanoceria may also have promising biomedical applications particularly in neurodegenerative and brain-related pathologies, but studies addressing their toxicity, and specifically on the nervous system, are still scarce, and their potential adverse effects and action mechanism are not totally understood yet. The objective of this work was to assess the biological behaviour of CeO NP in vitro in human nervous systems cells, and in vivo in Drosophila melanogaster to characterize their safety for exposed individuals and verify their suitability to be further employed in diagnosis and treatment of nervous system disorders. Cell cycle alterations, late apoptosis rate and DNA damage (comet and γH2AX assays), were determined in neuronal SH-SY5Y and glial A172 cells treated with nanoceria. Moreover, the survival rate, morphological changes and behavioural alterations were analysed in D. melanogaster individuals chronically exposed to CeO NP. The results obtained from the in vitro assessment showed that the nanoceria generally presented a good biocompatibility with scarce cyto- or genotoxic effects, essentially depending the exposure time and cell type, and being restricted to the longer exposure periods. Nevertheless, decrease in adult size and alterations observed in the larval crawling in the in vivo assays highlight the need of further investigations before establishing clinical uses of nanoceria.
- Two cinnamic acid derivatives as inhibitors of Pseudomonas aeruginosa las and pqs quorum-sensing systems: Impact on biofilm formation and virulence factorsPublication . Leitão, Miguel M.; Gonçalves, Ariana S.C.; Sousa, Sérgio F; Borges, Fernanda; Simões, Manuel; Borges, AnabelaIntroduction: Quorum sensing (QS) is a bacterial communication mechanism that regulates gene expression, playing a crucial role in various physiological processes. Interfering with this signalling pathway is a promising strategy to control bacterial pathogenicity and virulence. Objectives: This study evaluated the potential of two cinnamic acid derivatives, ferulic and sinapic acids, to inhibit the las and pqs systems in Pseudomonas aeruginosa. Their effects on biofilm architecture, virulence factor production and bacterial motility were also investigated. Methods: Bioreporter strains and bioluminescence-based assays were used to evaluate the modulation of QS-activity by cinnamic acid-type phenolic acids. In addition, in silico docking analysis was performed to validate the binding interactions of the cinnamic acid derivatives with QS-receptors. The biofilm architecture was analysed by optical coherence tomography, and virulence factors production (pyoverdine, pyocyanin, total proteases, lipases, gelatinases and siderophores) and motility were measured by absorbance measurement and plate agar method. Results: Ferulic and sinapic acids at 1000 µg mL-1 inhibited the las and pqs systems by 90 % and 80 %, respectively. The N-3-oxododecanoyl-homoserine lactone production was reduced by 70 % (6.25 µg mL-¹). In silico analysis demonstrated that cinnamic acid derivatives exhibited comparable interactions and higher docking scores than reference ligands and inhibitors. Biofilm thickness decreased from 96 µm to 11 µm, and virulence factors and swarming motility were significantly impaired. The comparable anti-QS activity of cinnamic acid derivatives suggests that the additional methoxy group in sinapic acid does not directly contribute to its anti-QS effect. Conclusion: Ferulic and sinapic acids compromised the biofilm architecture and virulence of P. aeruginosa through QS inhibition.
- An overview of work-related stress assessmentPublication . Lavreysen, Olivia; Bakusic, Jelena; Abatzi, Thalia-Anthi; Geerts, Annelien; Mateusen, Mies; Bashkin, Osnat; Koscec Bjelajac, Adrijana; Dopelt, Keren; du Prel, Jean-Baptist; Franic, Zrinka; Guseva Canu, Irina; Kiran, Sibel; Merisalu, Eda; Pereira, Cristiana Costa; Roquelaure, Yves; Godderis, LodeObjective: Work-related stress (WRS) is associated with the development of various health issues and long-term absence from the workplace. Adequate measurement of WRS is essential to assess its prevalence, risks, and effectiveness of preventive interventions. The aim of this review was to provide an overview of different categories of WRS assessment: 1) self-assessment, 2) external assessment, and 3) biomarkers. Methods: The databases MEDLINE, PsycINFO, EMBASE, CINAHL, and Web of Science have been searched until July 2024 for studies comprising self-assessment or external assessment of WRS, and WRS biomarkers. The self-assessment studies were further evaluated following the COSMIN guidelines. Results: In this review, a total of 15,749 articles were screened. The final analysis included 53 studies on self-assessment of WRS, 33 articles on external assessment of WRS and 167 articles on stress biomarkers. Within self-assessment studies, four instruments were included in the analysis: Job Content Questionnaire, Effort Reward Imbalance Questionnaire, Copenhagen Psychosocial Questionnaire II and the Demand-Control-Support Questionnaire. The studies applying external assessment used job-exposure matrices, work register data, ethnography, digital tools, and external observation. The identified WRS biomarkers were associated with the sympathetic adrenal medullary axis, the hypothalamic pituitary adrenal axis, immune response and inflammation, and haemostatic, metabolic and (epi)genetic biomarkers. Conclusion: The available evidence does not support the claim that there is a singular golden standard for assessing WRS. Inclusion of objective parameters and the interaction with subjective parameters and biological markers has to be studied to receive a broader view of WRS.
- New insights on antibacterial mode of action of blue-light photoactivated berberine and curcumin-antibiotic combinations against Staphylococcus aureusPublication . Gonçalves, Ariana S.C.; Fernandes, José R.; Saavedra, Maria José; Guimarães, Nuno M.; Pereira, Cristiana; Simões, Manuel; Borges, AnabelaAntimicrobial photodynamic inactivation (aPDI), using photosensitisers in combination with antibiotics, is a promising multi-target strategy to address antibiotic resistance, particularly in wound infections. This study aimed to elucidate the antibacterial mode of action of combinations of berberine (Ber) or curcumin (Cur) with selected antibiotics (Ber-Ab or Cur-Ab) under blue light irradiation (420 nm) against Staphylococcus aureus, including methicillin-resistant (MRSA) and methicillin-susceptible (MSSA) strains. Multiple physiological parameters were assessed using complementary assays (fluorometry, epifluorescence microscopy, flame emission and atomic absorption spectroscopy, zeta potential, flow cytometry, and the plate agar method) to examine the effect on ROS production, membrane integrity, DNA damage, motility and virulence factors of S. aureus. Results indicated that blue light photoactivated Ber-Ab and Cur-Ab combinations led to substantial ROS generation, even at low concentrations, causing oxidative stress that severely impacted bacterial membrane integrity (approximately 90 % in MRSA and 40 % in MSSA). Membrane destabilization was further confirmed by elevated intercellular potassium release (≈ 2.00 and 2.40 µg/mL in MRSA and MSSA, respectively). Furthermore, significant DNA damage was observed in both strains (≈ 50 %). aPDI treatment with blue light also reduced S. aureus pathogenicity by impairing motility and inhibiting key virulence factors such as proteases, lipases, and gelatinases, all of which play key roles in the infectious process. Overall, Ber-Ab combinations demonstrated the highest efficacy across all parameters tested, highlighting for the first time the multi-target therapeutic potential of this phytochemical-based aPDI strategy to combat antibiotic-resistant S. aureus infections and improve wound infection treatment outcomes.