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- 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.
- Association between cognitive reserve proxies and frailty phenotype: data from UK BiobankPublication . Lorenzo-López, Laura; Cibeira, Nuria; Hemadeh, Ali; López-López, Rocío; Lema-Arranz, Carlota; Maseda, Ana; Fernández-Bertólez, Natalia; Costa, Solange; Pásaro, Eduardo; Valdiglesias, Vanessa; Millán-Calenti, José C; Laffon, BlancaA potential protective role of cognitive reserve proxies against frailty has been suggested in older adults. We explored the cross-sectional association between cognitive reserve indicators and frailty phenotype. Data were obtained from the UK Biobank. We included 31,975 dementia-free participants aged ≥ 60 years (50.7% females, 2.2% frail) who completed a web-based cognitive assessment (fluid intelligence, working memory, visuospatial attention and processing speed, and executive functioning). Frailty was defined according to the Fried's phenotype (unintentional weight loss, exhaustion, low physical activity, slowness, and weakness). Participants meeting three or more criteria were classified as frail. Cognitive performance was compared between nonfrail and frail groups, and regression models were employed to analyze the associations between cognitive reserve proxies (education, skill level of occupation, social support, and multiple deprivation index (MDI)) and the likelihood of frailty. Frail and nonfrail groups significantly differed on cognitive function, with frail individuals demonstrating poorer performance on all cognitive functions (all p < .05) except fluid intelligence. Regression analysis showed that, after adjusting for age and sex, a lower educational level (odds ratio (OR) .797, 95% confidence interval (CI) .673-.944, p = .009), having maintained occupations with low cognitive requirements (OR .790, 95% CI .668-.936, p = .006), having less social support (OR .755, 95% CI .631-.903, p = .002), and living in a region with a high rate of multiple deprivation (OR 1.025, 95% CI 1.019-1.031, p < .001), significantly increased the probability of experiencing frailty. Our findings support the relationship between declined cognitive functions and frailty emphasizing the importance of implementing public health measures to enhance cognitive reserve.
- Association of inflammatory biomarkers with physical and cognitive frailty in a Spanish population of older adultsPublication . Lema-Arranz, Carlota; Hemadeh, Ali; Fernández-Bertólez, Natalia; Cibeira, Nuria; López-López, Rocío; Costa, Solange; Millán-Calenti, José Carlos; Lorenzo-López, Laura; Valdiglesias, Vanessa; Laffon, BlancaFrailty is a multifactorial geriatric syndrome characterized by increased vulnerability to stressors and associated with a higher risk of adverse health outcomes. Chronic low-grade inflammation has been proposed as a key pathophysiological mechanism underlying physical frailty, although its role in cognitive frailty remains undefined. In this cross-sectional study, we assessed the relationship between frailty status, both physical and cognitive, and plasma concentrations of six inflammatory biomarkers-C-reactive protein (CRP), interleukin 6 (IL-6), tumour necrosis factor alpha (TNF-α), soluble TNF-α receptor type II (sTNF-RII), high-temperature requirement serine protease A1 (HTRA1), and growth differentiation factor 15 (GDF15)-in a cohort of Spanish older adults (N = 150, ≥ 65 years old), classified according to Fried's frailty phenotype and frailty index. The results showed notable differences between frailty phenotype and frailty index, and highlighted CRP, TNF-α, sTNF-RII, and GDF15 as key biomarkers significantly associated with physical frailty status, with CRP and TNF-α also discriminating pre-frail individuals. sTNF-RII stood out for its high predictive capacity, while GDF15 added value as an indicator of sustained cellular stress. Regarding cognitive frailty, CRP, TNF-α, and GDF15 displayed significant associations with this condition. sTNF-RII and HTRA1, scarcely studied in this context, showed promising and significant associations (specific for cognitive frailty in the case of HTRA1) that justify their inclusion in future research aimed at better understanding the inflammatory mechanisms involved in cognitive frailty.
- Micronucleus assay in buccal and urothelial epithelial cells of wildland firefighters exposed to wildfire smokePublication . Esteves, Filipa; Madureira, Joana; Barros, Bela; Alves, Sara; Vaz, Josiana; Oliveira, Marta; Slezakova, Klara; Fernandes, Adília; do Carmo Pereira, Maria; Morais, Simone; Bonassi, Stefano; Teixeira, João Paulo; Costa, SolangeOccupational 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.
- Association between exposure to airborne endocrine disrupting chemicals and asthma in children or adolescents: A systematic review and meta-analysisPublication . Hatem, Georges; Faria, Ana Margarida; Pinto, Mariana Bessa; Teixeira, João Paulo; Salamova, Amina; Costa, Carla; Madureira, JoanaVarious studies have explored the association between Endocrine Disrupting Chemicals (EDCs) exposure and children's and adolescents' respiratory health, showing potential long-term effects and links to asthma. This systematic review explores the association between exposure to seven EDC groups during school age and adolescence and asthma onset or exacerbation while highlighting the predominant compounds underlying these potential associations. PubMed, Web of Science, Scopus, and Cochrane Library databases were searched with no time restriction. The risk of bias and heterogeneity across the included studies were assessed using the Newcastle Ottawa scale and I2 test, respectively. Pooled Odds Ratios (OR) and their 95% Confidence Intervals (CI) were calculated using the random effect model, and the quality of evidence for each outcome was measured using the GRADE approach. The review included 64851 children and adolescents from 61 observational studies, most with a low risk of bias in the studied domains. The pooled OR for asthma onset was significant for phthalates in dust samples (OR:1.21, CI:1.02; 1.44). Due to limited studies, the overall pooled effects for the other groups were not computed. Individual compounds demonstrating significant associations with asthma onset included airborne nickel (OR:1.10, CI:1.03; 1.18) and zinc (OR:1.13, CI:1.11; 1.15), urinary Bisphenol S (OR:1.40, CI:1.13; 1.73), Bisphenol A (OR:1.57, CI:1.02; 2.40) and arsenic (OR:2.08, CI:1.33; 3.26), and DiBP (OR:1.41, CI:1.08; 1.82), DEHP (OR:1.89, CI:1.00; 3.57), and TBOEP (OR:2.61, CI:1.08; 6.30) in the dust. Individual compounds with significant associations with greater asthma exacerbation odds comprised airborne nickel (OR:1.08, CI:1.01; 1.16) and zinc (OR:1.09, CI:1.01; 1.17), and urinary MEHHP (OR:1.24, CI:1.02; 1.51), MECPP (OR:1.30, CI:1.07; 1.57), MEOHP (OR:1.30, CI:1.09; 1.55), and MCOP (OR:1.32, CI:1.11; 1.57). Exposure to EDCs was significantly associated with asthma onset and exacerbation in children and adolescents, namely for phthalates, bisphenols A and S, arsenic, nickel, and zinc. Further research is recommended to focus on the impact of synergistic and co-exposure to other indoor air pollutants.
- Exposure to per-and poly-fluoroalkyl substances and respiratory and skin effects in children and adolescents: A systematic review and meta-analysisPublication . Hatem, Georges; Faria, Ana Margarida; Pinto, Mariana Bessa; Salamova, Amina; Teixeira, João Paulo; Costa, Carla; Madureira, JoanaDespite being previously banned due to long-term health effects, Per- and polyfluoroalkyl substances (PFAS) remain widespread in the environment, accumulating in animals and humans. This systematic review and meta-analysis explores associations between exposure to PFAS and asthma onset, wheezing, atopic dermatitis, and eczema in children and adolescents while addressing exposure timing and sex-specific differences. After comprehensive search conducted in several databases, including risk of bias, study heterogeneity, and quality of evidence evaluation, the review included 28 observational studies, most with low risk of bias in all domains. PFAS exposure was not significantly associated with asthma onset (OR:1.03, CI:0.99;1.07), but revealed significantly lower association in the prenatal period (OR:0.97, CI:0.94;0.99), higher in the postnatal period (OR:1.20, CI:1.07;1.35), and no differences among sexes. PFAS exposure (mainly prenatal) was associated with 4 % significantly lower odds of wheezing (OR:0.96, CI:0.94;0.98), higher in girls (OR:0.94, CI:0.91;0.98) than in boys (OR:0.97, CI:0.94;1.00). No significant impact was noted on atopic dermatitis (OR:1.04, CI:0.94;1.16), while PFAS exposure was associated with 8 % significantly lower eczema odds (OR:0.92, CI:0.89;0.96). Evidence was insufficient to perform sensitivity analyses on atopic dermatitis and eczema. Additional research is needed on the impact of synergistic and co-exposure to other pollutants on children and adolescents' health.
- Biocompatibility testing and antioxidant properties of cerium dioxide nanoparticles in human nervous system cellsPublication . Fernández-Bertólez, Natalia; Touzani, Assia; Ramos-Pan, Lucía; Reis, Ana Teresa; Teixeira, João Paulo; Laffon, Blanca; Valdiglesias, VanessaCerium dioxide nanoparticles (CeO NP), or nanoceria, are versatile materials with interesting properties for industry and medicine fields, particularly redox properties and catalytic activity. Because of their distinctive features, they have gained high attention in biomedical and pharmacological research to be employed in drug delivery, tissue regeneration, radioprotection, or diagnostic imaging. However, previous works reported that nanoceria may also induce reactive oxygen species (ROS) under certain conditions, leading to cellular stress, cellular damage, or cell death. In this study, the effects of CeO NP on cell viability and morphology as well as their influence on oxidative stress (both oxidant and ROS scavenging capacities) were investigated in nervous system cells (SH-SY5Y neuronal and A172 glial cells) treated with a wide range of CeO NP concentrations (1-100 µg/mL) for several treatment times. Results obtained showed that, despite being stable in time and effectively internalized by both cell types, CeO NP did not produce significant decrease in viability, evaluated by MTT assay, morphological alterations, or intrinsic cell-free ROS, but they generated cellular ROS limited to longer exposure periods. Furthermore, CeO NP demonstrated a certain intrinsic ability to scavenge ROS generated by HO in both tested cell types, more pronounced in neuronal cells. These results confirm the good biocompatibility of nanoceria on human nervous system cells and support further exploring their potential use in biomedicine field, particularly for those therapeutic and diagnostic applications related to the nervous system.