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- Clonal and plasmid-mediated dissemination of mcr-1 in Escherichia coli strains at the human–companion animal interface : genomic characterisation of colistin resistance plasmidsPublication . Menezes, Juliana; Silva, Joana Moreira da; Fernandes, Laura; Amaral, Andreia J.; Pomba, ConstançaThe global emergence of plasmid-mediated colistin resistance (mcr-1) gene poses a critical threat to human and animal health due to its ability for horizontal dissemination. While the role of food-producing animals is well recognised, the contribution of companion animals and household environments to the persistence and circulation of mcr-1 remains poorly understood. In this study, we investigated the genetic relatedness of mcr-1-positive Escherichia coli strains and their associated plasmids from dogs and their cohabiting humans in Portugal (2018–2020). Whole-genome sequencing, was performed on 17 strains, including repeated sampling from the same hosts over time. Core genome SNP analysis revealed clonal relatedness among several strains from the same host and between household members (≤6 SNPs). A total of 14 mcr-1-harbouring plasmids were identified and classified into three major incompatibility groups: IncX4 (n = 2), IncHI2 (n = 5), and IncI2 (n = 7). IncX4 plasmids were detected in clonally related strains from the same human host and were identical, indicating maintenance within a persistent lineage. A subset of IncI2 plasmids formed a closely related cluster (1–6 SNPs) across genetically distinct hosts, supporting the possibility of horizontal dissemination. IncHI2 plasmids displayed greater structural diversity and carried multiple antimicrobial and metal resistance determinants. Notably, chromosomal integration of mcr-1 was identified in three strains, suggesting a potential pathway for stabilistion of colistin resistance. Overall, these findings highlight the combined role of clonal expansion and plasmid circulation in shaping the epidemiology of mcr-1 genes in community settings, reinforcing the importance of genomic surveillance within a One Health framework.
- Clinical efficiency of lasers in endodontic treatment of primary endodontic cases : an umbrella review of systematic reviewsPublication . Duarte, Marta Alves; Neves, João Albernaz; Caçador, Madalena; Flores-Fraile, JavierObjectives: To evaluate and synthesise the current evidence on the efficacy of laser-activated irrigation in endodontics, with a focus on microbial reduction, postoperative outcomes, and its potential as an adjunct to conventional irrigants. Method: A comprehensive search was conducted to identify relevant systematic reviews. A total of 319 systematic reviews were initially identified. After the removal of 116 duplicates, 203 records were screened. Of these, 22 systematic reviews met the eligibility criteria. However, reviews that included exclusively in vitro studies or were considered to have limited clinical relevance were excluded. Ultimately, 9 systematic reviews were included in this umbrella review, evaluating different laser systems such as Er:YAG, Er,Cr:YSGG, Nd:YAG, and diode lasers in the context of root canal irrigation. This review was registered at PROSPERO (CRD42025599352). For this umbrella review, 5 electronic databases (PubMed/MEDLINE, Cochrane Database of Systematic Reviews, SciELO, Web of Science, and LILACS) were systematically searched from inception through December 2024, with no language or publication date restrictions applied. Study selection followed predefined eligibility criteria, and the methodological quality of the included systematic reviews was assessed using the AMSTAR 2 tool. Conclusions: The available evidence on laser-activated irrigation in endodontics remains limited and heterogeneous. Although some systematic reviews suggest potential benefits in canal disinfection and postoperative outcomes, the findings are inconsistent and largely constrained by variability in laser wavelengths, study designs, and outcome measures. Consequently, further standardised, high-quality clinical research is required before definitive conclusions regarding the clinical effectiveness of laser-activated irrigation can be established.
- Climatic sensitivity of migraine : a 14-year time series analysis of primary care consultations in SpainPublication . Cuenca-Zaldívar, Juan Nicolás; Villar, Carmen Corral del; Torres, Silvia García; Zamora, Rafael Araujo; Peña, Paula Gragera; Comte, Nina Cadeau; Almeida, André Mariz de; Sillevis, Rob; Sánchez-Romero, Eleuterio A.; Cid-Verdejo, RosanaBackground: Climatic variability has been proposed as a trigger for migraine; however, evidence from long-term primary care datasets remains scarce. Understanding how atmospheric conditions influence healthcare utilization may improve migraine prediction and management. This study aimed to analyze the association between climatic variables and weekly migraine consultations over a 14-year period in Spanish primary care and to identify the most accurate predictive time-series model. Methods: Weekly migraine consultations from 2010 to 2023 were extracted from electronic medical records using the International Classification of Primary Care, Second Edition (ICPC-2) code N89.01. Meteorological variables—temperature, diurnal variability, day-to-day change, wind direction and speed, barometric pressure, and sunshine hours—were obtained from the Spanish State Meteorological Agency (AEMET). Time-series analyses used exponential smoothing state-space models with external regressors (ETSX) and AutoRegressive Integrated Moving Average models with eXogenous regressors (ARIMAX). Model performance was assessed using Root Mean Squared Error (RMSE), Symmetric Mean Absolute Percentage Error (SMAPE), and Mean Absolute Scaled Error (MASE). Results: A total of 3176 migraine consultations were identified (mean age 47.6 ± 15.3 years; 81.7% female). The ARIMAX model showed the best predictive performance (RMSE = 3.485, SMAPE = 73.840, MASE = 0.875). Stationarity was confirmed using the Augmented Dickey–Fuller test (p = 0.01), and residuals showed no autocorrelation (Ljung–Box test, p = 0.833). After multivariable adjustment, female sex was the only variable independently associated with weekly migraine consultations; temperature, barometric pressure, diurnal variability, and wind speed showed no independent effects. Forecasting indicated a stable trend over the subsequent four years. Conclusions: This long-term time-series analysis showed that female sex was the only variable independently associated with weekly migraine consultations in primary care. Although most atmospheric indicators did not retain significance, climate-informed ARIMAX modeling improved prediction accuracy and may support personalized, weather-adapted preventive strategies.
- Chronic disease and disability in an 18th-Century Portuguese nun : an integrative multisource approachPublication . Antunes-Ferreira, NathalieThis study presents an integrative approach to chronic disease and disability in a Portuguese nun who died in 1779. The aim is to interpret her condition by combining osteopathological and burial context evidence with written sources. It offers a concise example of how bringing these sources together can enhance pathological interpretation and support the reconstruction of soft tissue disease, disability, and the lived experience of disease in past populations. This older woman was exhumed from the Convent of Jesus in Setúbal, Portugal. Her atypical burial position, also noted in the convent chronicles, indicates the remains likely belong to Michaela Archangela do Ceo. Osteopathological analysis revealed several changes, including atheromas, while the burial showed a crouched position, presumably related to stroke sequelae. The chronicles describe lifelong frailty and disease, followed in later life by a stroke with right-sided hemiplegia, pressure ulcers, and myiasis. Apart from the atheromas, which may be associated with the stroke, the skeletal changes show no correspondence with the signs reported in the account. The chronicles further document the attentive care she received, reflecting communal support within the convent, and the religious meaning of suffering. In conclusion, this case demonstrates the value of integrating multiple sources to interpret pathological conditions and the lived experience of disease, while emphasizing the relevance of soft tissue disease that leaves no skeletal trace. It also illustrates the careful application of retrospective diagnosis as an interpretive tool for historical documentation. Altogether, it highlights key methodological considerations for interpreting disease in osteoarchaeological contexts.
- CarieCheck : an mHealth app for caries-risk self-assessment : user-perceived usability and quality in a pilot studyPublication . Guerreiro, Eduardo; Souza, Guilherme; Mendes, José João; Manso, Ana Cristina; Botelho, JoãoBackground/Objectives: Mobile health (mHealth) technologies are increasingly used to support preventive oral care and patient self-management. CarieCheck is a Portuguese app intended to improve oral health literacy and support caries-risk self-assessment. This prospective pilot study focused on users’ perceived app quality and usability, assessed with uMARS-PT. Methods: Thirty participants from the academic community of Egas Moniz School of Health and Science used the app for 30 days and completed the uMARS-PT questionnaire. Descriptive statistics were used to calculate mean scores for Engagement, Functionality, Aesthetics, Information Quality, Subjective Quality, and Perceived Impact. Results: The overall mean uMARS-PT score was 4.22, indicating excellent perceived quality. The highest domain scores were Functionality (4.51), Aesthetics (4.45), and Information Quality (4.22). Engagement (3.71) and Subjective Quality (3.05) were moderate. Perceived Impact (3.85) reflected self-reported perception of increased awareness and motivation regarding oral health behaviors. Conclusions: CarieCheck was rated highly in usability, aesthetics, and information quality. These findings suggest that CarieCheck may be considered as a digital tool for preventive education and user-supported caries-risk self-assessment. Larger, longer-term studies in diverse populations using objective behavioral and clinical outcomes are warranted.
- Calcium phosphate bone substitutes in the prevention of bisphosphonate-related osteonecrosis of the jaw : a reviewPublication . Paulo, Siri; Abrantes, Ana Margarida; Laranjo, Mafalda; Marto, Carlos Miguel; Paula, Anabela; Trancoso, Pedro; Botelho, Filomena; Serra, Arménio; Ferreira, Manuel MarquesBisphosphonate-related osteonecrosis of the jaw (BRONJ) is characterized by exposed necrotic bone that often progresses with increasing pain and impaired quality of life. Zoledronate, the most potent and widely used bisphosphonate, has been strongly associated with BRONJ development following invasive dental procedures. Given the rising incidence of BRONJ, understanding and implementing effective preventive strategies have become imperative. Biomaterials based on synthetic hydroxyapatite and beta-tricalcium phosphate have been investigated as potential preventive agents. Their therapeutic rationale is supported by two key principles: the well-documented chemical interaction of calcium phosphates with bisphosphonates when used as drug carriers, and the established clinical use of synthetic calcium phosphate biomaterials in dentistry for bone regeneration. This review examines the underlying mechanisms of this preventive therapeutic strategy and evaluates studies investigating synthetic calcium phosphate biomaterials for BRONJ prevention through zoledronate adsorption at jaw wound sites, thereby reducing soft tissue toxicity and promoting healing. The evidence supports the protective effect of these biomaterials as a scientifically grounded preventive approach for BRONJ.
- Bovine ocular squamous cell carcinoma : a descriptive epidemiological survey in the Azores, PortugalPublication . Bilhastre, Beatriz; Vala, Helena; Ribeiro, Ana Clara; Faria, Sara; Oliveira, Ana; Branco, Sandra; Pinto, CarlosBovine ocular squamous cell carcinoma (BOSCC) is the most common ocular tumour in cattle, with a multifactorial aetiology involving ultraviolet (UV) radiation, genetic factors, pigmentation, and management practices. A detailed epidemiological characterisation of BOSCC in the Azores, Portugal, is provided, with particular emphasis on its spatial distribution and potential risk determinants. Data were obtained through an epidemiological questionnaire completed by field veterinarians between August 2023 and March 2025. A total of 85 BOSCC cases were recorded across 62 farms—45 on Terceira Island and 17 on São Miguel Island. All affected animals were adult Holstein Friesian dairy cows, managed under extensive pasture-based systems. The nictitating membrane was the most frequently affected structure (69.5%), and multiple lesions occurred in 20% of the cases. Farms located at 200–400 m of altitude presented the highest number of cases. Continuous exposure to UV under pasture-based management represents the main environmental risk factor. Although periocular pigmentation may provide partial protection, other environmental and genetic factors can also contribute to tumour development. Artificial insemination is considered a promising preventive tool, enabling genetic selection for protective traits such as periocular pigmentation. This research provides the first regional epidemiological characterization of BOSCC in the Azores, highlighting the interplay among environmental, genetic, and management-related factors in disease occurrence.
- Botulinum toxin type A for the prevention of migraines : an umbrella review of systematic reviewsPublication . Chamani, Goli; Jasim, Hajer; Minston, Ava; Dias, Marlon Ferreira; Poluha, Rodrigo Lorenzi; Gonçalves, Daniela A. Godoi; Christidis, Maria; Al-Moraissi, Essam Ahmed; Christidis, Nikolaos; Canales, Giancarlo De la Torre; Ernberg, MalinBotulinum toxin type A (BoNT-A) is an established preventive therapy for chronic migraines; however, uncertainty remains regarding its comparative efficacy and safety. Thus, we aimed to summarize current evidence from high-quality systematic reviews of the therapeutic effects of BoNT-A in migraine management. An umbrella review was conducted following PRISMA guidelines and registered in PROSPERO. High-quality systematic reviews with meta-analysis evaluating BoNT-A efficacy were identified through five databases up to August 2024. Primary outcomes included monthly headache frequency and severity. Methodological quality and risk of bias were assessed using the umbrella review checklist. Fourteen articles were included. Overall, quantitative evidence indicated favorable effects of BoNT-A compared with placebo for chronic migraines, across headache frequency, headache severity, and acute medication use, but less efficacy than topiramate and the CGRP monoclonal antibodies (CGRPmAbs) galcanezumab and fremanezumab. Though the adverse events were frequent, BoNT-A was generally well-tolerated. Comparative data suggested superior tolerability versus topiramate and a safety profile like CGRPmAbs. Although botulinum toxin type A is widely used as a preventive treatment for chronic migraines, the available evidence supports its efficacy at a moderate level. Further head-to-head and long-term analyses are needed to clarify its comparative role alongside newer biologic treatments.
- Bone availability in the sagittal dimension of the infrazygomatic crest for miniscrew insertion : a retrospective cone-beam computed tomography studyPublication . Furão, Susana; Bugaighis, Iman; Proença, Luís; Brito, José; Costa, Hélder Nunes; Pereira, François Durand; Pereira, Pedro MarianoBackground: There is limited information available regarding the bone volume in the sagittal dimension of the infrazygomatic crest (IZC) that is safe for the insertion of a temporary skeletal anchorage device. The objective of this study was to assess the sagittal bone availability when the anterior and posterior walls of the IZC reach an inclination of 45°. Methods: A cross-sectional investigation was undertaken on 100 cone-beam computed tomography scans of 40 males and 60 females. A total of 20 landmarks were digitized for each IZC outline, 1 mm from the vestibular alveolar bone, and their x, y, and z coordinates were extracted. Afterward, the sagittal measurement of the IZC at which the curvature reached 45° was recorded. Symmetry of measurements was assessed using paired-samples t tests. In addition, these measurements were compared as a function of sex and age group (≤21 vs >21 years) using an independent-samples t test and the Spearman rank correlation coefficient test (P <0.05). Results: There were no significant differences between paired sagittal measurements of the IZC (3.5 ± 1.5 mm on the right side and 3.6 ± 1.3 mm on the left side; P = 0.144), except for younger patients (P = 0.030). In addition, no significant differences were found between corresponding male and female measurements (P ≥0.149). A significant discrepancy was identified between the younger age group (3.2 ± 1.2 mm) and the older cohort (3.8 ± 1.6 mm) for the sagittal dimension of the right IZC (P = 0.038), with an estimated mean difference of 0.03-1.16 mm (95% confidence interval). A significant positive weak correlation was found between the sagittal dimension of the right IZC and age (ρ = 0.201; P = 0.045). Conclusions: Sufficient IZC bone volume is available at an angle of 45° for inserting a temporary skeletal anchorage device, with no sex or side variation, except for the significantly greater sagittal dimension of the right IZC in older patients.
- Bioengineered baculovirus-derived extracellular vesicles loaded with of γ-carboxylated Gla-rich protein : dual modulation of inflammation and vascular calcificationPublication . Viegas, Carla; Pichard, Simon; Carreira, Joana; Ova, Adélia; Troffer-Charlier, Nathalie; Maia, Teresa M.; Edelweiss, Evelina; Macedo, Anjos L.; Matos, António; Faria, Tiago Q.; Calado, Sofia M.; Monico, Carina; Devos, Simon; Impens, Francis; Schaeffer-Reiss, Christine; Cianférani, Sarah; Peixoto, Cristina; Poterszman, Arnaud; Simes, DinaChronic inflammation and ectopic calcification are interrelated processes driving major chronic inflammatory diseases such as cardiovascular and chronic kidney diseases. Gla-rich protein (GRP), a vitamin K–dependent protein (VKDP) with dual anti-inflammatory and anti-calcific properties, has emerged as a promising therapeutic molecule. However, its biomedical development has been limited by difficulties in producing the γ-carboxylated (cGRP) form and by its poor solubility at physiological pH, constraining formulation and delivery. To address these challenges, we established a baculovirus expression vector system (BEVS) designed to couple GRP post-translational maturation with its secretion in extracellular vesicles (EVs). Co-expression of GRP with γ-glutamyl carboxylase (GGCX), vitamin K epoxide reductase (VKOR), and the convertase Furin enabled efficient γ-carboxylation, propeptide removal, and secretion of mature cGRP. GGCX and VKOR were essential for γ-carboxylation, while Furin mediated propeptide processing. EVs were isolated by differential ultracentrifugation into 30 K and 100 K fractions and characterized by NTA, TEM, Western blot, ELISA, and proteomics. All vesicles displayed physical and molecular features resembling mammalian EVs, including canonical EV markers and distinct proteomic profiles, with GRP, GGCX, VKOR, and Furin preferentially enriched in the 30 K population. Functional assays demonstrated that the resulting EVs associated with human THP-1 macrophages and vascular smooth muscle cells (VSMCs) without inducing cytotoxicity, and both cGRP-EVs and uncarboxylated GRP-EVs reduced pro-inflammatory cytokine release while exerting dual anti-inflammatory and anti-mineralizing effects. This study establishes the first bioengineered platform capable of generating functional γ-carboxylated GRP and its vesicular formulation, providing a dual innovation for VKDP research and therapeutic biomaterial development.