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- An approach to quality control of the dithiothreitol (DTT) assay for the determination of oxidative potential of atmospheric particulate matterPublication . Vicente, Carolina; Gonçalves, Sara; Gamelas, Carla; Almeida, Susana Marta; Canha, NunoThe oxidative potential (OP) of airborne particulate matter (PM) has emerged as a promising metric to assess the capacity of particles to induce oxidative stress and related health effects. Thus, ensuring the reliability and comparability of OP measurements is essential for accurate environmental and toxicological assessment. This study aimed to develop and evaluate a quality control approach for the dithiothreitol (DTT) assay used in OP determination. The DTT assay provides an estimation of how harmful PM can be to human health through oxidative stress, relating it to the consumption of DTT during the test period. Two experiments were conducted using the Standard Reference Material (SRM) 1648–Urban Particulate Matter (NIST, USA). The first assessed the effect of trichloroacetic acid (TCA) addition order and the feasibility of using SRM 1648 as a reference material. The second evaluated the stability of the SRM solution over 63 days. Statistical analysis (Mann–Whitney test) indicated that the order of TCA addition did not significantly affect OP values (p > 0.05). SRM 1648 solution determination showed high reproducibility (mean DTTₘ = 14.6 ± 2.4 pmol·min−1·µg−1), although a gradual increase in DTT metrics was observed over time, consistent with progressive dissolution. The results support the application of SRM 1648 as a reference material for DTT assay quality control, supporting methodological harmonization in OP determination, provided that a freshly prepared solution is used.
- Factors associated with the recurrence of intermittent exotropia and reoperations in the long termPublication . Lino, Pedro; Aguiar, Pedro; Cunha, João PauloPurpose: To evaluate long-term reoperation risk after bilateral lateral rectus (BLR) recession for intermittent exotropia (IXT) in a paediatric cohort, and to identify the perioperative factors associated with recurrence and reoperation. Methods: A retrospective observational cohort of 258 children with basic or divergence-excess IXT who underwent BLR recession at CUF Cascais Hospital between 2010 and 2020 was analysed. Clinical variables included age, age at surgery, pre- and immediate postoperative deviation angles, preoperative occlusion therapy, orthoptic treatment, binocular function, and initial surgical success (residual deviation <10 prism dioptres with fusion). Bivariate analyses were performed using t-tests or χ2 tests, and variables with p < 0.10 were entered into multivariable binary logistic regression to identify the independent predictors of reoperation. Model diagnostics included variance inflation factors (VIF), Hosmer-Lemeshow test, and area under the receiver operating characteristic curve (AUC). Significance was set at p < 0.05. Results: Reoperation was required in 11% of cases. Failed initial surgery (adjusted OR = 34.7; p < 0.001), larger preoperative deviation (adjusted OR per PD = 1.143; 95% CI 1.062-1.231; p < 0.001), larger immediate postoperative deviation (adjusted OR per PD = 0.822; 95% CI 0.766-0.881; p < 0.001), and older patient age (adjusted OR per year = 1.261; 95% CI 1.032-1.542; p = 0.024) were independently associated with reoperation. Orthoptic and occlusion therapies showed non-significant trends. Conclusion: In children undergoing BLR for IXT, initial surgical failure and deviation magnitude (pre- and immediate postoperative) are the strongest predictors of reoperation, with age having a modest additional effect. These findings emphasize precise surgical planning and the need for long-term follow-up.
- Old molecules, new hope: a scoping review and bibliometric analysis of drug repurposing for lung cancerPublication . Ragassi, Wellington Martins; Alves, Fernando Miguel; Lazo, Raul Edison; Tonin, Fernanda; Pontarolo, Roberto; Sari, Marcel Henrique; Ferreira, Luana MotaDrug repurposing has gained prominence in oncology by enabling the investigation of approved drugs for new therapeutic purposes. In lung cancer, this strategy may reduce the time and costs associated with drug development. This study aimed to map the landscape of in silico, in vitro, in vivo, and clinical research on drug repurposing for lung cancer, while identifying key molecular targets and research gaps. A systematic search was conducted in PubMed, Embase, and Web of Science, following Joanna Briggs Institute and PRISMA-ScR guidelines. Two reviewers independently selected and extracted the data. A total of 58 studies, published between 2010 and 2024, mainly from the United Kingdom (19%) and the United States (17%), were included. Most studies used in vitro models (53%), followed by in vivo (31%) and in silico (16%), with frequent combinations of methods. The most investigated drug classes were antibiotics (10%), antipsychotics (9%), antidiabetics (8%), anthelmintics (6%), and antihistamines (6%). Frequently studied drugs included niclosamide, metformin, atorvastatin, and doxazosin, targeting pathways such as PI3K/AKT/mTOR, apoptosis, and autophagy. Bibliometric analysis revealed increasing scientific output, with emphasis on combination therapies, cellular mechanisms, and technologies like molecular docking and nanosystems. These findings highlight the growing relevance of drug repurposing in lung cancer, especially in accelerating effective therapy discovery using approved compounds. Progress in this field depends on integrating diverse methodologies and fostering interdisciplinary collaboration. As a next step, rigorous clinical trials are essential to confirm the efficacy and safety of promising repurposed agents in oncology.
- Aspergillus spp. in woodworking settings: implications for occupational health and safetyPublication . Dias, Marta; Rodriguez, Margarida; Riesenberger, Bruna; Marques, Liliana; Carolino, Elisabete; Twarużek, Magdalena; Kosicki, Robert; Gonçalves, Lídia; Caetano, Liliana Aranha; Alastruey-Izquierdo, Ana; Viegas, Susana; Viegas, CarlaWoodworkers are exposed to several potentially harmful agents, including microorganisms that grow in the wood. The most common fungal species in woodworking environments are Aspergillus, Penicillium, and Cladosporium spp., with occupational exposure to Aspergillus spp. posing a significant respiratory risk. This study aimed to assess exposure to Aspergillus spp. in two Portuguese woodworking environments and to perform a thorough analysis of Aspergillus fumigatus complex isolates collected from 13 DIY stores and 6 Carpentries in the Lisbon Metropolitan Area. Sampling combined active and passive methods to assess microbial contamination. Aspergillus fumigatus isolates were analysed for their antifungal susceptibility, resistant mechanisms, mycotoxin production, and cytotoxic potential in lung (A459) and liver (HepG2) cell lines. The MAS-100 presented Aspergillus sections Aspergilli and Flavi with the highest prevalence in DIY stores and Carpentries, respectively. A total of 1185 Aspergillus spp. were recovered, 270 identified as Aspergillus fumigatus sensu stricto growing at 37 °C. None of those isolates was resistant to azoles, 99.07% of them produced gliotoxin, and 39.9% of them produced cytotoxic effects in at least one cell line. This study comprehended a multi-approach that considered not only sampling methods but also the laboratory assays to be applied in the Aspergillus section Fumigati isolates recovered from two different woodworking environments, allowing a complete and robust analysis of this specific environment and species. Overall, the findings indicate that woodworkers are exposed to A. fumigatus isolates with relevant pathogenic traits, despite the absence of azole resistance, underscoring the need for continued environmental and occupational monitoring.
- The indoor microbiome: sampling, analysis and emerging trendsPublication . Šunić, Iva; Šarac, Jelena; Auguštin, Dubravka Havaš; Pozdniakova, Sofya; Ferguson, Robert M.; Jergović, Matijana; Visentin, David; Borràs, Silvia; Archer, Elizabeth; Henderson, Drew K.; Vitko, Sandra; Ašić, Adna; Bošnjaković, Anja; Maglica, Željka; Viegas, Carla; Novokmet, Natalija; Karlović, Nina; Marjanović, Damir; Muszyński, Adam; Liu, Yuxi; Karisola, Piia; Alenius, Harri; Krych, Lukasz; Lovrić, MarioIndoor spaces contain diverse microbial communities that shape human health. These microorganisms are particularly relevant to respiratory diseases, including asthma and allergies. Despite growing recognition of the importance of indoor microbial exposures, research in this field is slowed by differences in methods. These inconsistencies make it difficult to compare results and draw conclusions. This systematic review analyses 106 studies published between 2000 and 2025 that investigated indoor microbiomes in dust, air, and other matrices across homes, schools, and other built environments. We assessed sampling strategies, DNA extraction protocols, sequencing technologies, and bioinformatic pipelines, identifying trends, inconsistencies, and areas requiring harmonisation. Passive sampling, particularly dust collection, was the most common approach, while Illumina-based 16S rRNA and ITS amplicon sequencing dominated molecular analyses. However, variations in targeted gene regions, extraction kits, and analytical tools limited cross-study comparability. Ecological findings revealed consistent detection of bacterial taxa such as Staphylococcus, Streptococcus, and Corynebacterium, and fungal taxa including Cladosporium, Aspergillus, and Penicillium, with diversity shaped by building characteristics, ventilation, humidity, occupancy, and presence of pets. This review highlights the need for standardised protocols in indoor microbiome research to facilitate reproducibility, enable meta-analyses, and inform health-related guidelines for indoor environments.
- Neoplasia maligna com características de adenocarcinoma extrauterino: estudo de caso clínicoPublication . Babo, A.; Silva, J.; Roque, RubenAs neoplasias malignas identificadas em citologia cervicovaginal com características de adenocarcinomas extrauterinos são neoplasias raras e agressivas, frequentemente diagnosticadas em estadios avançados, devido à sua apresentação clínica pouco específica. Este tipo de carcinoma representa um desafio significativo para o diagnóstico, exigindo uma abordagem interdisciplinar. Neste estudo, apresentamos um caso clínico de uma mulher de 75 anos com extensa carcinomatose peritoneal de predomínio infraumbilical. A citologia cervicovaginal em meio líquido revelou uma neoplasia maligna compatível com adenocarcinoma extrauterino, mas a origem exata não pode ser determinada com certeza. O exame histológico do tumor revelou tratar-se de um carcinoma misto com componente de carcinoma seroso de alto grau e um componente de carcinoma de células claras, não fazendo distinção entre ovário/trompa de Falópio/peritoneu ou origem endometrial. Este caso salienta a capacidade de deteção da citologia em neoplasias extrauterinas, enfatizando a importância crítica da integração de técnicas complementares para a caracterização e diagnóstico definitivo destes carcinomas.
- Metastização leptomeníngea de origem ginecológica: o papel decisivo da citologiaPublication . Dias, C.; Lino, I.; Babo, M.; Roque, RubenO exame citopatológico do líquido cefalorraquidiano é um método diagnóstico essencial na avaliação de patologias do sistema nervoso central, sendo fundamental na deteção de disseminação meníngea por neoplasias. O presente caso de estudo refere-se a uma mulher de 34 anos previamente diagnosticada com carcinoma adenoescamoso do colo do útero associado ao vírus do papiloma humano de alto risco, com metastização ganglionar confirmada. Cerca de um ano após histerectomia radical, perante sintomatologia neurológica, foi equacionada a hipótese de leptomeningite carcinomatosa. A análise citológica do líquido cefalorraquidiano revelou a existência de células malignas de morfologia epitelial, confirmada por imunocitoquímica, com imunomarcação positiva para a pancitoqueratina MNF116, a proteína p16 e negativa para CD20. A história oncológica prévia e a morfologia celular, complementada pela avaliação de biomarcadores por testes de imunocitoquímica, permitiram o diagnóstico de metastização leptomeníngea de carcinoma do colo do útero. Este caso evidencia a relevância do exame citológico do líquido cefalorraquidiano que, associado a técnicas complementares, permitiu o diagnóstico de metástases meníngeas de neoplasia de origem ginecológica, entidade rara, mas clinicamente relevante.
- Carcinoma medular da tiroide com neoplasia papilar síncrona: metástase ganglionar inaugural diagnosticada por citologia aspirativaPublication . Araújo, A.; Pires, Carolina; Oliveira, M.; Roque, RubenA citologia aspirativa por agulha fina (CAAF) é um método essencial na caracterização das adenopatias cervicais, permitindo a distinção entre lesões benignas e malignas. O carcinoma medular da tiroide, uma neoplasia neuroendócrina rara, derivada das células C da tiroide, pode apresentar-se inicialmente como metástase ganglionar. Este artigo descreve o caso clínico de uma mulher de 62 anos com adenomegalia supraclavicular, cuja primeira CAAF foi sugestiva de neoplasia epitelial sem origem definida. Posteriormente, realizou nova CAAF, o que suscitou a hipótese de metástase de carcinoma medular da tiroide. A confirmação diagnóstica foi obtida por técnicas de imunocitoquímica (avaliação dos biomarcadores TTF1 e calcitonina). A tiroidectomia total com esvaziamento cervical revelou um carcinoma medular no lobo direito, com invasão vascular, e um microcarcinoma papilar folicular no lobo esquerdo. Este caso consubstancia o papel da citologia na abordagem inicial das adenopatias, evidenciando a importância da CAAF na obtenção de amostras representativas e com preservação morfológica adequada, mesmo sendo um método minimamente invasivo. Aliada a um processamento eficaz e acompanhada por técnicas complementares, revela-se fundamental para um diagnóstico citopatológico exato e para a correta orientação clínica.
- The potential of DNA methylation as a biomarker for age-related macular degeneration: a systematic reviewPublication . Ghannai, Salema; Ribeiro, Edna; Pereira, Bruno; Brito, Miguel; Camacho, PedroAge-related macular degeneration (AMD) is a multifactorial disease influenced by genetic and environmental factors, yet its pathogenesis remains incompletely understood. DNA methylation, increasingly recognized as a disease indicator, has been linked to AMD and may represent a promising biomarker or therapeutic target. This systematic review, conducted according to PRISMA 2020 guidelines, analyzed 13 studies addressing DNA methylation in AMD populations (2012-2025). Results revealed that 25% reported hypermethylation, 8% hypomethylation, and 41% both patterns, while 15% found no significant differences. Notably, one study described downregulation of DNA methyltransferases in advanced stages compared with early or intermediate AMD. Despite the limited evidence, findings support the relevance of methylation in AMD prognosis and therapy. Further research with robust methodologies is essential to clarify the role of epigenetic mechanisms in disease progression and to explore their potential for guiding targeted therapeutic strategies
- The comet assay as a tool in human biomonitoring exposure to anaesthetic gases: a systematic review and meta-analysisPublication . Gajski, Goran; Haveric, Anja; Møller, Peter; Azqueta, Amaya; Giovannelli, Lisa; Gerić, Marko; Stopper, Helga; Bankoglu, Ezgi Eyluel; Collins, Andrew; Ladeira, CarinaAnaesthetic gases are agents used to induce and maintain general anaesthesia during surgical procedures. Common examples include sevoflurane, isoflurane, and desflurane, which act by depressing the central nervous system to produce unconsciousness and analgesia. These gases are administered through a vaporiser and inhaled via a mask or endotracheal tube. While effective, they can contribute to environmental pollution and increase the risk of occupational exposure. Medical personnel working in operating or post-operative facilities are unavoidably exposed to anaesthetic gases. Several adverse health effects have been associated with anaesthetic gas exposure; therefore, this review aims to summarise findings on DNA strand breaks, assessed by the comet assay in leucocytes of exposed medical workers. Standardised mean differences (SMDs) have been calculated by random effects models. The meta-analysis included 16 studies. Of these, 11 showed statistically significant increased levels of DNA strand breaks, whereas another five studies showed no significant effect. Overall, there is an increased level of DNA strand breaks in exposed subjects in unadjusted analysis (SMD = 1.17, 95 % confidence interval: 0.71, 1.62) as well as analysis adjusted for missing studies by the trim-and-fill method (SMD = 0.53, 95 % confidence interval: −0.14, 1.21). In conclusion, this systematic review and meta-analysis demonstrate that exposure to anaesthetic gases in an occupational setting induces primary DNA damage in human leucocytes, warranting further research to minimise any adverse effects on exposed medical personnel. Besides, the relevance of the use of the comet assay in assessing DNA damage in human biomonitoring studies is proven.