Search Results (785)

Background: The athletic potential of young athletes is shaped by individual and environmental factors. Objectives: This study examines the physical growth, body composition, biological maturation, motivation, perseverance, physical performance and contextual factors of young male and female track and field athletes. Methods: A total of 425 (224 girls) track and field athletes were recruited and divided into five age cohorts (10, 11, 12, 13, and 14 years respectively). Measurements were assessed across (i) individual (anthropometry, body composition, biological maturation, motivation, and perseverance), (ii) performance (motor performance), and (iii) club context domains. Data analysis used descriptive statistics for clubs’ characteristics, a two-factor ANOVA for anthropometry, body composition, biological maturation, and performance and an ANCOVA for motivation and perseverance. All analyses used STATA 18.0. Results: Sex-related differences were identified in physical growth, maturation, psychological, and performance variables during adolescence. Girls reached their peak height velocity (PHV) around 12 years of age, compared to 14 years in boys. At all ages (except at age 11), girls had higher body fat, and at age 12 were taller and outperformed boys in right-handgrip strength and in sprint (30 m and 40 m). From age 13 years onwards, boys became taller, with greater leg length, greater fat-free mass, and superior results (p < 0.05) in most performance tests. Psychologically, girls reported higher levels of interest–enjoyment, effort–importance, relatedness, and perceived choice; no sex differences were found in perseverance. The clubs involved were of small size, with developing, yet qualified, coaches, with limited support staff and infrastructure. Conclusions: Clear sex differences in physical growth, psychological, and performance variables emerged during adolescence, and were related in part to earlier maturation in girls. Further, there was variation in clubs’ infrastructure and staff that may potentially influence track and field athletes’ growth and development. Full article

Background/Objectives: Self-care is a cornerstone of healthy aging and chronic disease management; however, evidence on the most effective intervention models for improving quality of life in older adults with chronic non-communicable diseases (NCDs) remains fragmented. This review aimed to evaluate the effectiveness of self-care interventions in promoting quality of life and health outcomes in older adults with NCDs. Methods: A systematic review was conducted in accordance with PRISMA 2020 guidelines and registered in PROSPERO (CRD420251040613). Randomized and non-randomized clinical trials published between 2019 and 2024 were retrieved from Scopus, Web of Science, and EBSCOhost. Eligible studies included adults aged ≥60 years with NCDs receiving self-care interventions. Data extraction and risk of bias assessment were independently performed using Joanna Briggs Institute tools. Results: Twenty-nine studies involving 7241 older adults were included. Self-care interventions comprised nurse-led educational programs, digital health strategies, community- and peer-based approaches, and person-centered care models. Multicomponent and continuous interventions demonstrated consistent improvements in physical and psychological domains of quality of life, self-efficacy, autonomy, symptom management, and treatment adherence. Digital interventions enhanced monitoring and engagement, although their effectiveness varied according to sensory and health literacy limitations. Conclusions: Structured, person-centered, and nurse-led self-care interventions are effective in improving quality of life and autonomy among older adults with NCDs. These findings support their integration into primary and community-based care, reinforcing their relevance for clinical practice, care planning, and the development of assistive and educational strategies in aging care. Full article

The adoption of conservation systems in agriculture has been increasingly explored as a strategy to improve soil quality and potentially influence greenhouse gas (GHG) emissions. This study reports the first assessment of GHG emissions within a long-term (14 years) agroecological field experiment evaluating soil management systems for onion (Allium cepa L.) production in a Humic Dystrudept (Cambissolo Húmico Distrófico, Brazilian Soil Classification System) in Southern Brazil. Three management systems based on permanent soil cover and crop diversification were evaluated in an onion–maize rotation: conventional tillage (CT) without cover crops, no-till (NT) without cover crops, and a no-till vegetable system (NTV) with a summer cover crop mixture of pearl millet (Pennisetum americanum), velvet bean (Mucuna aterrima), and sunflower (Helianthus annuus). Short-term GHG emissions were monitored during one onion growing season (106 days), while soil chemical and physical properties reflect long-term management effects. Evaluations included (i) daily and cumulative GHG (N₂O, CH₄, and CO₂) emissions, (ii) soil carbon (C) and nitrogen (N) stocks, (iii) soil aggregation, porosity, and bulk density in different soil layers (0.00–0.05, 0.05–0.10, and 0.10–0.30 m), and (iv) onion yield and cover crop dry matter production. The NTV system improved soil physical and chemical quality and increased onion yield compared to NT and CT. However, higher cumulative N₂O emissions were observed in NTV, highlighting a short-term trade-off between increased N₂O emissions and long-term improvements in soil quality and crop productivity. All systems acted as methane sinks, with greater CH₄ uptake under NTV. Despite higher short-term emissions, the NTV system maintained a positive C balance due to long-term C accumulation in soil. Short-term greenhouse gas emissions were assessed during a single onion growing season, whereas soil carbon stocks reflect long-term management effects; CO₂ fluxes measured using static chambers represent ecosystem respiration rather than net ecosystem carbon balance. These results provide an initial baseline of GHG dynamics within a long-term agroecological system and support future multi-year assessments aimed at refining mitigation strategies in diversified vegetable production systems. Full article

Candida spp. are important opportunistic human fungal pathogens. This study aimed to identify and characterize Candida spp. obtained from patients admitted to an Intensive Care Unit (ICU), focusing on virulence attributes and susceptibility to antifungal agents. A total of 131 isolates from oral and tracheobronchial secretions of adult ICU patients were evaluated. Phenotypic identification was performed using chromogenic culture media for Candida, followed by MALDI-TOF mass spectrometry, with representative isolates confirmed by ITS sequencing. Antifungal susceptibility to fluconazole, ketoconazole, and amphotericin B was determined only by the agar disk diffusion method, and virulence was assessed through esterase, DNase, protease, and hemolytic activity assays. C. albicans was the prevalent species, followed by C. tropicalis, C. krusei, C. glabrata, C. parapsilosis, C. dubliniensis, C. lusitaniae, and C. guilliermondii. Antifungal resistance rates reached 51.1% for fluconazole, 42.7% for ketoconazole, and 19.1% for amphotericin B, as determined by disk diffusion method. Overall, 64.9% of the isolates exhibited esterase activity, 18.3% DNase, 45.8% protease, and 67.2% exhibited hemolytic activity. Oral isolates were more frequent than tracheal isolates and demonstrated a higher prevalence of antifungal resistance and virulence traits. These findings underscore the epidemiological importance of characterizing Candida species in hospitals to better understand the yeast profile and to support adequate clinical management. Full article

The aim of this study was to evaluate different doses of bacterial inoculants and the inclusion of 8% ground corn grain (GCG) on fermentative characteristics, chemical composition, and in situ ruminal degradability of low-DM elephant grass (cv. BRS Capiaçu) silage. The experiment followed a completely randomized design in a 6 × 3 factorial arrangement (six treatments × three fermentation periods). Treatments were a control without additive (CTR); 0.5 or 1 g/ton of Lentilactobacillus buchneri (LBU0.5 and LBU1); 1 or 2 g/ton of a Lactiplantibacillus plantarum + Pediococcus acidilactici inoculant (LPP1 and LPP2); and 8% GCG. After 60 d of fermentation, in situ ruminal degradability was evaluated using rumen-fistulated lactating cows with incubation times from 0 to 240 h. The GCG treatment increased DM, CP, and ether extract concentrations and reduced NDF, ADF, and lignin contents. Additionally, GCG silage exhibited lower pH, butyric acid, and ammonia nitrogen concentrations, along with higher lactic acid levels. No treatment effects were observed for water-soluble carbohydrates or total DM losses. The effective NDF degradability, degradation rate of the slowly degradable fraction, and undigested NDF after 240 h were not affected by treatments. In conclusion, the inclusion of GCG improved the fermentative profile of low-DM elephant grass silage, whereas bacterial inoculants did not significantly enhance the silage quality under the conditions evaluated. Full article

Background and Objectives: Obesity has been increasing sharply worldwide and is related to diabetes, cardiovascular diseases, liver disease, and cancer. Sleeve gastrectomy is the most used surgical approach to reduce body weight and treat metabolic implications observed in patients with moderate-to-severe obesity. On the other hand, this procedure affects the musculoskeletal system, and investigating skeletal muscle is not routinely recommended for bariatric surgery. This study aimed to evaluate the psoas muscle in patients in the preoperative period of sleeve gastrectomy and six months after the procedure using abdominal computed tomography scans. Materials and Methods: This clinical, exploratory, and observational study, with a prospective longitudinal observational study design, was conducted at a single center with 31 women who underwent sleeve gastrectomy. The evaluations were performed before and after six months of the procedures. Results: Anthropometric, muscle strength, hepatic ultrasound, and psoas computerized tomography evaluations were performed. A significant reduction in body weight, body mass index, waist, neck, and calf circumference was observed. There was also a substantial reduction in right-hand strength and the area and index of the psoas muscle (but with an increase in density). Most presented a routine abdominal ultrasound. Conclusions: Our results suggest that muscle evaluation provides valuable information for clinical monitoring before and after bariatric surgery, helping to identify potential risks and guide multidisciplinary follow-up. Psoas muscle area and psoas muscle index decreased, but psoas muscle density increased, all significantly. These results indicate that conducting a muscle evaluation is helpful for patients undergoing bariatric surgery, supporting the use of the clinical approach before and after the procedure, predicting possible complications, and providing more accurate prognoses. Full article

Type 2 diabetes mellitus (T2DM) involves chronic hyperglycemia, insulin resistance, low-grade inflammation, and oxidative stress that drive cardiometabolic and renal damage despite current therapies. Sodium–glucose cotransporter (SGLT) inhibitors have reshaped the treatment landscape, but residual risk and safety concerns highlight the need for new agents that combine glucose-lowering efficacy with redox–inflammatory modulation. LASSBio-1986 is a synthetic N-acylhydrazone (NAH) derivative designed as a gliflozin-like scaffold with the potential to interact with SGLT1/2 while also influencing oxidative and inflammatory pathways. Here, we integrated in silico and in vivo approaches to characterize LASSBio-1986 as a multifunctional antidiabetic lead in murine models of glucose dysregulation. PASS and target class prediction suggested a broad activity spectrum and highlighted transporter- and stress-related pathways. Molecular docking indicated high-affinity binding to both SGLT1 and SGLT2, with a modest energetic preference for SGLT2, and ADME/Tox predictions supported favorable oral drug-likeness. In vivo, intraperitoneal LASSBio-1986 improved oral glucose tolerance and reduced glycemic excursions in an acute glucose challenge model in C57BL/6 mice, while enhancing hepatic and skeletal muscle glycogen stores. In a dexamethasone-induced insulin-resistance model, LASSBio-1986 improved insulin sensitivity, favorably modulated serum lipids, attenuated thiobarbituric acid-reactive substances (TBARS), restored reduced glutathione (GSH) levels, and rebalanced pro- and anti-inflammatory cytokines in metabolic tissues, with efficacy broadly comparable to dapagliflozin. These convergent findings support LASSBio-1986 as a preclinical, multimodal lead that targets SGLT-dependent glucose handling while mitigating oxidative and inflammatory stress in models relevant to T2DM. Chronic disease models, formal toxicology, and pharmacokinetic studies, particularly with oral dosing, will be essential to define its translational potential. Full article

Background/Objectives: Patients in need of specialized palliative care are clinically highly complex, with pain being the most prevalent problem. Furthermore, in these patients, a self-report for characterization of pain could be difficult to obtain. This cross-sectional, exploratory study investigates the use of clinical parameters and peripheral blood biomarkers for potentially identifying and characterizing pain (assessed using Pain Assessment in Advanced Dementia (PAINAD) and Numeric Scale (NS)) in patients under palliative care, including a population with dementia where pain is often underdiagnosed. Methods: Fifty-three patients with non-oncological diseases were analyzed in a cross-sectional study using medical and nursing records. Among previous biomarkers related to monocytes and platelets assessed by flow cytometry, we selected the most significative ones for pain characterization in a logistic regression analysis (multivariate analysis), alongside patient-specific characteristics such as renal function, nutritional status, and age. Results: Our exploratory findings suggest strong relationships between chronic pain and advanced age, reduced glomerular filtration rate (GFR), and malnutrition within this cohort. Furthermore, the percentage of lymphocytes, total and classical monocytes, the relative expression in monocytes of CD206, CD163, the CD163/CD206 ratio, and the relative expression in platelets of CD59 emerged as potential predictors of pain. Statistical analyses highlighted the challenges of multicollinearity among variables such as age, GFR, and nutritional status. A classification model further suggested that all patients over 65 years in our specific sample reported pain. Conclusions: This pilot study provides preliminary support for prior evidence linking chronic pain to aging, nutritional deficits, and renal impairment, and highlights potential novel peripheral blood biomarkers for pain assessment. This work emphasizes the promise of clinical and molecular biomarkers to improve pain detection and management, contributing to personalized and effective palliative care strategies. Full article

Alternative energies are essential for meeting the global demand for environmentally friendly energy, especially as the use of fossil fuels is being reduced. In recent years, largely due to diminishing fossil fuel reserves, Portugal has been actively promoting investment in renewable energies to reduce its reliance on energy imports and fossil fuels. However, despite the country’s high daily sunshine hours and utilization of wind and hydropower, energy production remains unstable due to climate variability. Climate instability leads to fluctuations in the energy supplied to the grid and can even partially withstand blackouts such as the one that occurred on 28 April 2025 on the Iberian Peninsula. To address this problem, energy storage systems are crucial to guarantee the stability of the supply during periods of low production or in situations such as the one mentioned above. This paper analyzes the feasibility of implementing an energy storage system to increase the profitability of a wind farm located in Alto Douro, Portugal. The study begins with a demand analysis, followed by simulations of the system’s performance in terms of profitability based on efficiency and power. Based on these assumptions, a modular lithium battery storage system with high efficiency and rapid charge/discharge capabilities was selected. This battery, with less autonomy but high capacity, is more profitable, since a 5% increase in efficiency results in high profits (€84,838) and curtailment (€70,962) using batteries with lower autonomy, i.e., 2 h (power rating of 5 MW combined with 10 MWh energy storage). Therefore, two scenarios (A and B) were considered, with one more optimistic (A) in which the priority is to discharge the batteries whenever possible. In the more realistic scenario (B), it is assumed that the batteries are fully charged before discharge. On the other hand, in the event of a blackout, it enables faster commissioning of the surrounding water installations, because solar and battery energy have no inertia, which facilitates the back start protocol. Full article

This study evaluated the effects of different bentonite levels on the fermentation profile, losses, microbial populations, aerobic stability, and chemical composition of corn silage. A 5 × 3 factorial completely randomized design was used, with five bentonite inclusion levels (0, 20, 40, 60, and 80 g/kg) and three storage times (30, 90, and 180 days), with four repetitions. Significant additive × storage time interactions were detected for key parameters. After 180 days, pH increased to 4.20 at 80 g/kg bentonite, while lactic acid peaked at 54.02 g/kg DM with 60 g/kg at 90 days. Acetic acid reached 22.36 g/kg DM at 180 days, and lactic acid bacteria ranged from 3.42 to 7.43 log CFU/g. Yeast counts were lowest (≤0.57 log CFU/g) at 180 days with 20–60 g/kg bentonite. Dry matter rose to 311 g/kg with 60 g/kg bentonite, whereas soluble carbohydrates decreased from 123.9 to 68.2 g/kg DM. Aerobic stability markedly improved, reaching 142.06 h with 40 g/kg bentonite at 180 days, almost three times higher than the control. The inclusion of bentonite in corn silage at levels of 20–60 g/kg of fresh matter effectively enhances silage quality during 180 days of storage by improving fermentation characteristics, microbial stability, and aerobic stability. Full article

This paper presents the rationale and design of a study of growth and development in young track and field athletes: the EXcellence and PERformance in Track and field (EXPERT) study, and details the methodologies used. Background: Longitudinal research examining individual-environment interactions in youth athletic development is scarce for track and field. Objectives: The EXPERT study investigates how individual (anthropometry, maturation, motivation) and environmental (family, coach, club) characteristics influence developmental trajectories in youth track and field athletes. Methods: A mixed-longitudinal design will follow 400 athletes (200♂, 200♀; aged 10–14 years) from 40 Portuguese clubs across five cohorts assessed biannually over three years. Guided by Bronfenbrenner’s bioecological model, assessments encompass individual, performance, health, and environmental domains. Data quality control will consist of rigorous training of all research team members, implementation of standardized protocols, a pilot study, and an in-field reliability study. Multilevel growth models will examine trajectories and predictor effects of predictors. Conclusions: EXPERT will provide evidence to optimize training and support holistic youth athlete development. Full article

Background/Objectives: The biogenic synthesis of silver nanoparticles (AgNPs) is a promising alternative method, driven by the presence of metabolites in plant matrices capable of acting as reducing and stabilizing agents. Seasonality is a key factor that influences the phytochemical composition of plants and can directly impact the yield, physicochemical characteristics, stability, and bioactivities of the obtained AgNPs. This study aimed to synthesize AgNPs using aqueous extracts from Paullinia cupana leaves collected during dry and rainy seasons, prepared by two different methods (agitation or infusion), to evaluate the impact of these variables on the biosynthesis and properties of the nanostructures. Methods: The extracts were characterized by UHPLC-HRMS/MS, and their total phenolic compound (TPC) content and antioxidant potential against DPPH and ABTS radicals were determined. The AgNPs were characterized by UV/Vis spectrophotometry, dynamic light scattering (DLS), zeta potential (ZP), nano-particle tracking analysis (NTA), transmission electron microscopy (TEM), and energy-dispersive X-ray spectroscopy (EDX). Results: The metabolic profile results showed a predominance of alkaloids and flavonoids in all extracts, with greater phytochemical diversity in samples prepared by infusion. TPC indicated superior phenolic extraction in extracts prepared by infusion during the rainy season, correlating with greater antioxidant potential via the elimination of free radicals. The evolution of AgNP synthesis was accompanied by a gradual change in the color of the suspensions and the formation of plasmon bands between 410 and 430 nm, characteristic of spherical AgNPs. The nanostructures presented hydrodynamic diameters between 37.49 and 145.5 nm, PdI between 0.222 and 0.755, and Zeta potential between −11.3 and −39.9 mV, suggesting satisfactory colloidal stability. Morphological analyses revealed predominantly spherical particles with average diameters ranging from 33.61 to 48.86 nm and uniform distribution, while EDX spectra confirmed the presence of silver. Conclusions: Thus, our results demonstrate that both seasonality and the method of extract preparation influence the phytochemical composition and, consequently, the morphology, stability, and optical properties of AgNPs, with subtle emphasis on collections made during the rainy season and extracts prepared by infusion. Such knowledge contributes to the advancement of more reproducible and purpose-oriented syntheses in the field of green nanotechnology, enabling applications in various sectors. Full article

Background: Oncological treatment remains a major clinical challenge. Despite therapeutic advances and the diversity of available approaches, many tumors continue to exhibit limited responsiveness to chemotherapy. In this context, nutrition has emerged as a promising complementary strategy to support cancer therapy. In particular, interventions based on nutritional deprivation have gained prominence due to their ability to modulate tumor metabolism, inducing alterations that may increase the sensitivity of cancer cells to conventional treatments. Accordingly, the present study aimed to evaluate the safety and efficacy of caloric restriction combined with chemotherapy in a Sarcoma-180 model, investigating its effects on immunological and hematological parameters, antioxidant activity, oxidative stress, and tumor and liver morphology, as well as DNA damage. Methods: Mice bearing Sarcoma-180 were randomly assigned to four groups: Ad Libitum (AL), Ad Libitum + Doxorubicin (ALDOX), Caloric Restriction (CR), and Caloric Restriction + Doxorubicin (CRDOX). Assessment included tumor weight and volume, food and caloric intake, hematotoxicity, lipid metabolism, oxidative stress and antioxidant markers, genotoxicity, morphological alterations in the tumor and liver, and overall survival. Results: The data obtained demonstrate that caloric restriction combined with doxorubicin is both safe and feasible, as it preserves body weight and does not induce metabolic disturbances. Importantly, this combined strategy produced a marked reduction in tumor volume and mass while also mitigating the hematotoxicity typically associated with doxorubicin. In peripheral blood, the regimen decreased chemotherapy-induced DNA damage, supporting a systemic protective effect. Consistently, the combination reduced oxidative stress markers (NOx and MDA) and enhanced antioxidant activity within the tumor. Histological analyses further confirmed these outcomes, showing tumor cell death with features compatible with apoptosis and reduced local invasion. Together, these data indicate that caloric restriction enhances the antitumor efficacy of doxorubicin while simultaneously improving treatment tolerance. Conclusions: This study demonstrates that caloric restriction, combined with doxorubicin, is safe, well-tolerated, and enhances the antitumor response in the Sarcoma-180 model. Full article

Background: Real-world safety profiles of immune checkpoint inhibitors (ICIs) in older adults remain insufficiently characterized. Although ICIs are widely used across tumor types, older patients, particularly those with frailty, multimorbidity, or polypharmacy, are consistently under-represented in clinical trials, limiting the external validity of trial-derived toxicity estimates. Robust real-world data are therefore essential to clarify the incidence, seriousness, and age-related patterns of immune-related adverse events (irAEs) in routine practice. Methods: This is a nationwide retrospective study of spontaneous ICI-related ADRs reported in INFARMED’s Portal RAM (2011–2024). We evaluated the frequency, seriousness, fatality, and organ-specific patterns of ICI-related adverse drug reactions (ADRs) reported to the Portuguese National Pharmacovigilance System. The analytic unit was the ADR case. Endpoints included seriousness (primary), fatality, hospitalization, time-to-onset, and System Organ Class. Multivariable logistic regression adjusted for age, sex, regimen, tumor type, polypharmacy, and calendar period; sensitivity analyses using first ADR per patient were concordant. Results: We identified 2300 eligible ICI-related ADRs (corresponding to 925 patients). Median age at the time of ADR was 65 years (IQR not reported); 33.7% occurred in adults aged ≥70 years, and 62.8% of reports involved male patients. PD-1 inhibitors accounted for 77.5% of ADRs, and monotherapy for 72.9%. Overall, 85.8% of ADRs were classified as serious; 17.9% led to hospitalization and 19.1% were fatal. Serious-event reporting was similar in older and younger adults (≥70 vs. <70 years: 84.5% vs. 86.5%, p = 0.22), and the proportion explicitly labeled immune-related did not differ (9.3% vs. 8.7%, p = 0.56). In contrast, fatal outcomes were significantly more common in older adults (25.3% vs. 16.0%; p < 0.001). Age was associated with distinct organ-specific patterns. Adults ≥ 70 years had higher odds of nervous system disorders (aOR 1.75, 95% CI 1.23–2.48) and immune system disorders (aOR 1.42, 95% CI 1.02–1.98), but lower odds of hepatobiliary (aOR 0.52, 95% CI 0.36–0.76; p = 0.001) and blood/lymphatic disorders (aOR 0.50, 95% CI 0.32–0.79). In multivariable models, age ≥ 70 years did not predict seriousness (aOR 0.98, 95% CI 0.76–1.27), whereas combination therapy remained independently associated with increased seriousness (aOR 1.57, 95% CI 1.13–2.18). Conversely, age ≥ 70 years independently predicted fatal outcomes (aOR 1.66, 95% CI 1.31–2.09). Later calendar periods (2017–2024) were associated with substantially lower fatality (aOR 0.16; 95% CI 0.10–0.27). CTLA-4-containing regimens demonstrated a tendency toward higher fatality (aOR 1.50; 95% CI 0.94–2.37). Conclusions: Chronological age does not seem to increase the likelihood of reporting a serious ICI-related ADR, but, once toxicity occurs, older adults experience higher fatality rates. Age-related phenotypic differences and regimen-specific risks highlight the need for early recognition systems and tailored toxicity management in older populations. Full article

Aflatoxins (AFs) are potent hepatotropic mycotoxins—AFB₁ being the best-characterized—yet their ability to induce intrahepatic cholangiocarcinoma (iCCA) remains underexplored. Male Wistar rats received vehicle (controls; n = 5) or an AFB₁-dominant AF mixture (AFB₁ 39.46 μg/mL; AFB₂ 1.13 μg/mL; AFG₁ 17.44 μg/mL; AFG₂ 0.59 μg/mL— n = 10) by daily gavage for 90 days, at a dose equivalent to 400 μg AFB₁ per kg of diet. After 12 months, twelve iCCA tumors were resected and analyzed by histology (H&E) and tissue-microarray-based immunohistochemistry (Cytokeratin-19, Hep Par-1, p53, Cyclin D1, Rb, β-catenin, and PCNA). Lesions predominantly showed glandular/tubular architecture consistent with iCCA and were cytokeratin-19-positive and Hep Par-1-negative. Cell proliferation was high (PCNA ≈ 69%). p53 displayed nuclear accumulation in 83% of tumors. G1/S control was perturbed, with cyclin D1 overexpression (67%), and Rb was positive in 58% of iCCA. Aberrant Wnt activation was rare (nuclear β-catenin in 8%). Subchronic exposure to an AFB₁-dominant AF mixture in rats was associated with iCCA characterized by high proliferative activity, p53 accumulation, and disruption of G1/S checkpoint components. These findings broaden the oncogenic spectrum of AFs and warrant genomic confirmation of AF mutational signatures. Full article