Hospitais da Universidade de Coimbra

: Flow cytometry is a powerful tool for immunophenotyping, but its application in feline samples is challenging due to species-specific blood characteristics, a paucity of standardised protocols and high reagent costs. These limitations may compromise sample quality, antibody performance, and the reproducibility of the results. : An optimal protocol for extracellular immunophenotyping of feline leukocytes from peripheral whole-blood was developed, adapting established human flow cytometry methods. Key adaptations included improved blood collection process, assessing sample preservation, and titrating antibodies. Detailed MIFlowCyt-compliant information on cytometer configuration, compensation procedures, gating strategy, and controls is provided in Supplementary File S2. : The enhanced blood collection significantly improved erythrocyte lysis rendering the samples more suitable for cytometric analysis. Baseline leukocytes viability, assessed by trypan blue exclusion was 98-100%. The use of a cellular antigen stabilisation reagent preserved feline peripheral whole-blood samples without detectable loss of surface antigen expression for up to 14 days. Antibody titration showed that most monoclonal antibodies were effective at 1.5 µL, reducing usage by up to 85%, while CD5 required 3 µL. : This improved, cost-effective and reproducible protocol address major technical limitations in feline flow cytometry and provides a practical framework for the reliable leukocyte immunophenotyping in clinical diagnostics, research, and comparative immunology.

ST-segment elevation myocardial infarction (STEMI) is a life-threatening condition needing urgent reperfusion, with better outcomes linked to optimal myocardial perfusion. Statins may reduce ischemia-reperfusion injury through pleiotropic effects. While the 2019 European Society of Cardiology (ESC) Dislipidemia Guidelines recommended high-intensity statins before percutaneous coronary intervention (PCI), this was not included in the 2023 ESC or 2025 ACC/AHA/ACEP/NAEMSP/SCAI ACS Guidelines. To evaluate the effect of high-dose statin loading before primary PCI in improving coronary reperfusion and clinical outcomes in STEMI patients through a systematic review and meta-analysis of randomized controlled trials (RCTs). On June 2025, CENTRAL, EMBASE, and PubMed were searched for RCTs comparing high-dose statin preloading with placebo or no statin in STEMI patients undergoing primary PCI. The primary outcome was post-PCI thrombolysis in myocardial infarction (TIMI) flow grade 3. Secondary outcomes included all-cause mortality and greater than or equal to 50% ST-segment resolution. Data were pooled using random-effects models, reporting odds ratios (OR) with 95% confidence intervals (CIs). Nine RCTs with 1862 patients were included. High-dose statin preloading significantly increased the likelihood of achieving post-PCI TIMI flow grade 3 (OR: 1.52, 95% CI: 1.06-2.20). It was also associated with a reduction in all-cause mortality (OR: 0.59, 95% CI: 0.36-0.98). No significant difference was observed in greater than or equal to 50% ST-segment resolution (OR: 1.04, 95% CI: 0.71-1.50). High-dose statin loading before primary PCI in STEMI patients seems to be associated with improved coronary reperfusion and reduced all-cause mortality. These findings offer timely and clinically relevant insight, reminding the cardiovascular community of the potential value of this therapeutic.

Background: Attention deficit and hyperactivity disorder (ADHD) is linked to a higher risk of injuries, especially in childhood and adolescence. This study compares emergency department (ED) visits between pediatric patients with ADHD and healthy controls, identifying potential risk factors and differences in injury severity. Methods: Retrospective cohort study including patients diagnosed with ADHD at a Neurodevelopment Clinic, and a control group of patients born at the same hospital. Medical records were reviewed for clinical data (number of ED accidents since birth and severity outcomes such as splints, hospitalization, surgeries, inter-hospital transfers and sequelae). Parametric measurements were used due to the sample size and absence of skewness. Negative binomial regression accounted for overdispersion in incidence rates. Results: We analyzed 316 ADHD cases and 316 controls. Mean age was 11.9 years (SD 2.4), similar to controls (p=0.998), with 66.5% males in both groups. ADHD patients had 537 unintentional injury ED visits (controls: 387), an incidence rate ratio (IRR) of 1.36 (95% CI: 1.11-1.67). The IRR was higher for episodes ≥6 years (IRR 1.48; 95% CI: 1.16-1.88). Severity outcomes were similar (p=0.257). Lower parental education was associated with more episodes (IRR 1.39; 95% CI: 0.989-1.96). Comorbidities did not affect injury incidence (p=0.425), but patients without comorbidities required more procedures and transfers (p=0.018 and p=0.013, respectively). 79.7% of cases had most/all ED visits before starting psychostimulants. Conclusions: Our study supports the increased risk of injuries among pediatric ADHD patients. While injury severity was comparable to controls, ADHD patients without comorbidities had greater severity. Psychostimulants suggested a protective effect.

Introduction: Intracranial stenting during endovascular thrombectomy (EVT) is a common practice in the setting of failed reperfusion or severe stenosis. Immediate stent patency requires periprocedural antiplatelet therapy (APT). How APT intensity interacts with prior intravenous thrombolysis (IVT) to influence hemorrhagic risk remains uncertain. We aimed to assess whether the APT regimen modifies the association of IVT with early intracranial hemorrhage after intracranial stenting during EVT. Methods: This was a subanalysis of the RESISTANT registry, a multicenter, international, retrospective cohort (2016 to 2023) of adults with acute ischemic stroke who underwent intracranial stenting during EVT. APT regimens were categorized as conservative (intravenous or oral aspirin alone, or aspirin plus an oral P2Y12 inhibitor) and aggressive (any regimen including intravenous GPIIb/IIIa inhibitor or intravenous cangrelor). Four main groups were compared according to the APT regimen (conservative/aggressive) and the use of IVT (+/-). The primary outcome was a composite of sICH and parenchymal hematoma types 1 and 2 (sICH-PH2-PH1). Multivariable logistic regression models were used to evaluate the interaction between IVT and APT, adjusting for clinically relevant covariates. Results: Among the 823 included patients, 44 (5.3%) received conservative APT with IVT, 130 (15.8%) received conservative APT without IVT, 145 (17.6%) received aggressive APT with IVT, and 504 (61.2%) received aggressive APT without IVT. Among patients who received IVT, sICH-PH2-PH1 rates were 9.3% with conservative APT and 10.7% with aggressive APT; among those without IVT, rates were 3.2% and 9.9%, respectively. Administration of IVT (adjusted odds ratio [aOR] 5.84, 95%CI 1.07 to 43.92; p=0.05) and aggressive APT (aOR 4.81, 95% CI 1.41 to 30.22; p=0.03) were each associated with higher odds of hemorrhagic complications, with a significant IVT by APT interaction (P interaction =0.05; Figures 1 and 2 ). Within the aggressive APT plus IVT subgroup, sICH-PH2-PH1 occurred in 20% of patients treated with cangrelor and 6.1% treated with a glycoprotein IIb/IIIa inhibitor ( Figure 3 ). Conclusion: Among patients requiring intracranial stenting, aggressive periprocedural APT and prior IVT are each associated with higher hemorrhagic risk, with the combination showing the worst observed crude outcome. Prospective evaluation of protocolized APT pathways in the IVT setting is warranted.

Metastatic urothelial carcinoma (mUC) is a lethal cancer with limited therapeutic options. Advances in genomic and transcriptomic research have deepened the understanding of mUC biology, leading to the identification of clinically relevant molecular alterations that represent potential actionable targets. This has broadened the treatment landscape of the disease to include novel agents, such as antibody-drug conjugates (e.g., enfortumab vedotin) and targeted therapies, including the pan-fibroblast growth factor receptor (FGFR) inhibitor erdafitinib. Genomic alterations in FGFR3 are well-established oncogenic drivers in bladder cancer and represent predictive biomarkers of response to FGFR-targeted therapies. The phase III THOR trial demonstrated the clinical benefit of erdafitinib in previously treated mUC patients harboring FGFR3 alterations and supported its subsequent approval by the European Medicines Agency. In this context, accurate molecular profiling is essential to guide patient selection for FGFR inhibitor therapy. Equally important is the standardization and timely implementation of FGFR3 testing in clinical practice to optimize treatment planning. This review addresses key considerations in FGFR3 testing in mUC and discusses how it can be routinely incorporated into clinical practice.

Self-regulation is the basic capacity to regulate one's thoughts, emotions, and behaviors. The aim of the present study is to examine the psychometric properties of the Self-Regulation Scale (SRS) among male and female Portuguese youth participants (N = 559 youth, M = 16.51 years, SD = 1.07, range = 14-20 years). The three-factor model composed of the Emotional, Cognitive, and Behavioral regulation subscales obtained adequate fit, although the fit of the second-order model was also acceptable. Internal consistency as measured by the alpha and omega reliability estimators was good. Significant associations were found with psychometric measures of relevant constructs (e.g., self-control, psychopathic traits, criminogenic cognitions), and external criterion-related variables (e.g., engaging in activities against the law, alcohol and drug abuse). Cross-gender measurement invariance was supported, with females scoring significantly higher on the Cognitive regulation subscale, and males scoring significantly higher on the Emotional regulation subscale. The findings support the use of the SRS to validly and reliably measure self-regulation in Portuguese youth.

Epicardial adipose tissue (EAT) function may influence the heart, given its metabolic actions and proximity to the heart. We hypothesized that diabetes mellitus (DM) alters miRNA expression across adipose tissue types, and that modifications in EAT may have critical implications for cardiac physiology. To test this, we compared EAT and subcutaneous adipose tissue (SAT) miRNA profiles between patients with and without DM and across tissues within each disease group. Paired biopsies from patients with (n = 18) and without DM (n = 46) undergoing cardiac surgery were analyzed using miRNA profiling and bioinformatics. Among 680 miRNAs screened, 34 were uniquely expressed in EAT, confirming a distinct molecular signature in this fat depot. Notably, miR-155-5p was significantly elevated in EAT from patients with DM, indicating a localized metabolic effect. In SAT, miR-93-3p and miR-223-3p were upregulated in patients with DM and consistently higher than in EAT, regardless of DM status, indicating tissue-specific regulation. miR-324-5p was more expressed in SAT of patients in the NDM group, reflecting combined effects of tissue type and DM. These patterns remained consistent across cardiac disease stratifications. Pathway analysis revealed that miRNAs enriched in EAT target genes involved in cardiomyocyte growth and differentiation. Overall, the findings highlight the unique miRNA profile of epicardial fat and its altered response to DM, supporting its relevance in cardiac physiology.

Despite widespread use of tumor necrosis factor inhibitors (TNFi) as first-line therapy in rheumatoid arthritis (RA), up to 40% of patients fail initial treatment. Subsequent therapeutic choices remain poorly structured, with limited evidence-based guidance to inform individualized post-TNFi decision-making. To develop evidence-informed, profile-based recommendations to guide treatment selection after inadequate response to a first TNFi in RA, combining evidence and expert consensus. Delphi-based consensus study informed by a PRISMA-guided scoping review (ScR) and nominal group methodology. A PRISMA-guided ScR of biologic and targeted synthetic biologic disease-modifying antirheumatic drug (tsDMARDs) after TNFi failure was conducted. Patient profiles were identified by a steering committee, and draft recommendations were evaluated through an anonymized Delphi process. A profile-based decision tree integrated direct and indirect evidence, with evidence strength graded using the Oxford Centre for Evidence-Based Medicine approach. The ScR included 43 studies, mostly exploratory analyses of randomized trials. Scenarios included age ⩾65 years; failure of ⩾2 TNFi; monotherapy; rheumatoid factor/anti-citrullinated peptide antibody status; prominent systemic inflammation; interstitial lung disease (ILD); rheumatoid vasculitis; high cardiovascular (CV) risk or prior CV event; venous thromboembolism (VTE) risk; obesity; high infection risk; osteoporosis; nociplastic pain, depression and fatigue; prior solid cancer; hematologic cancer/lymphoproliferative disease; non-melanoma skin cancer; and pregnancy.Seventeen recommendations were formulated; 15 achieved consensus. Agreed positions included caution with JAK inhibitors (JAKi) in older patients and in those with CV/VTE risk; preference for IL-6 receptor inhibitors or JAKi for monotherapy or prominent systemic inflammation; in RA-ILD, use a b/tsDMARD with a non-TNFi mechanism; rituximab as first choice in rheumatoid vasculitis; abatacept in infection-prone patients; discouraging JAKi in prior malignancy; and TNFi as acceptable during pregnancy. Two statements did not reach consensus: preferential use of non-TNFi in obesity and heightened caution with tofacitinib in osteoporosis or fracture risk. This Delphi-validated, profile-based framework provides a practical tool to support evidence-informed clinical decision-making.