Search Results (16,430)

This study evaluated the effects of the dietary administration of a glycosylated form of active vitamin D (calcitriol, 1,25(OH)₂D₃) combined with ursolic acid (UA) and oleanolic acid (OA) triterpenes on sow health and productivity. Twenty-four third-parity Landrace × Large White sows were allocated at day 108 of gestation into three groups: a control group receiving 1800 IU/kg of vitamin D₃, and two treatment groups receiving the control diet supplemented with either 0.64 µg/kg (ACTD1) or 0.96 µg/kg (ACTD2) of glycosylated 1,25(OH)₂D₃ plus 140 or 210 µg/kg of UA + OA (4:1 ratio), respectively. Diets were administered from late gestation through the end of lactation. Farrowing duration, sow body weight, backfat thickness, and litter growth were recorded. Blood samples collected at key physiological stages were analyzed for pro-inflammatory cytokines, mineral homeostasis, endocrine markers, and serum proteome. Farrowing time was reduced in both treatment groups compared with the control (p < 0.05). Treated sows exhibited lower backfat thickness at the end of lactation and improved litter weights at farrowing, after cross-fostering, and at weaning (p < 0.05). Plasma pro-inflammatory cytokines (TNF-α, IL-1α, and IL-1β) were reduced at the end of lactation in ACTD1 and ACTD2 sows, with TNF-α and IL-1β already decreased after farrowing (p < 0.05). Treated sows also displayed decreased plasma parathormone concentrations at the end of lactation, along with increased circulating 1,25(OH)₂D₃ and calcium concentrations after farrowing and at lactation end (p < 0.05), while plasma phosphate levels remained unchanged. Proteomic analysis supported the systemic availability of the supplemented compounds and their involvement in metabolic and inflammatory pathways rather than calcium transport or vitamin D binding mechanisms. Overall, this nutritional strategy influenced the immune modulation while maintaining mineral homeostasis via modest endocrine adaptations. Larger-scale trials are warranted to confirm these results and to evaluate their practical applicability under commercial production conditions. Full article

Objectives: Dendrobine (DDB) is one of the active ingredients in Dendrobium and has been reported to have significant neuroprotective properties. Nevertheless, the precise mechanisms underlying its action have not been fully clarified. The microglial imbalance of polarization is regarded as one of the key determinants in the etiology of neurodegenerative conditions, in the contribution of neuroinflammation. The recovery of M1/M2 balance and the inhibition of over-production of the pro-inflammatory effects have become major topics in modern studies of preventing and treating neurodegenerative diseases. Methods: Therefore, the present study aimed to explore the effects of DDB on the Lipopolysaccharide (LPS)-induced neuroinflammatory model in BV2 microglial cells and the potential molecular mechanisms of microglial M1/M2 polarization. Result: The results showed that DDB significantly suppressed Nitric Oxide (NO) release and ROS levels in LPS-induced BV2 cells. ELISA, qPCR, Western blot, and immunofluorescence results indicated that DDB reduced pro-inflammatory mediators Tumor Necrosis Factor-alpha (TNF-α), Interleukin-6 (IL-6), and nterleukin-1 beta (IL-1β) and increased anti-inflammatory mediators Interleukin-10 (IL-10) and Arginase-1 (Arg-1). Consistently, it decreased M1-like markers Inducible Nitric Oxide Synthase (iNOS) and Cluster of Differentiation 16/32 (CD16/32) while increasing M2-like/repair-associated markers (CD206 and Arg-1), suggesting a shift toward a more anti-inflammatory microglial activation profile based on the assessed marker pane. Conclusions: These results suggested that DDB can suppress the production of inflammatory cytokines and modulate microglial polarization, which indicated that DDB can be used as an effective compound in the prevention of neuroinflammation-related disorders. Full article

Treponema pallidum is the etiological cause of syphilis, and in recent years, reemergence has been reported, especially among men who have sex with men and people living with HIV (PLWH). Certain cytokines may act as hallmark biomarkers in the progression of syphilis in PLWH, and studying how the immune system works against T. pallidum is important, especially in PLWH, whose immune system is compromised. We evaluated the serum expressions of IFN, TNF, IL-10, TGF-β1 and IL-17 in men living with HIV (MLWH) and their association with distinct stages of syphilis. We recruited MLWH from March to October 2022. A blood sample was requested, syphilis was detected using the reverse algorithm, and antibodies were titrated to determine the stage. Each of the cytokines studied was quantified using commercial ELISA kits. The following groups were formed: active syphilis (n = 217), cured syphilis (n = 134), and without syphilis (n = 159). The prevalence of elevated TGF-β1 differed between groups, being highest in individuals with active syphilis (51.6%; median 319 pg/mL), followed by those with cured syphilis (41.0%; median 137.0 pg/mL). Younger participants and persons without a history of sexually transmitted infections were more likely to present with high TGF-β1 levels. TGF-β1 may act as a biomarker in active syphilis and could suppress the inflammatory response against spirochetes. Full article

Background: Both chronic kidney disease (CKD) and the haemodialysis procedure can contribute to disturbances in mineral homeostasis, which can potentially result in cellular pathologies. Our study aims to investigate trace element levels in haemodialysis patients and evaluate their potential impact on cellular adhesion molecules. This will clarify the clinical significance of trace element imbalances in this population. Methods: The study included 84 haemodialysis patients and 42 healthy controls. Trace element levels in blood (Zn, Cu, Mn, Mo, V, Sb and Cr) were measured using inductively coupled plasma mass spectrometry (ICP-MS), and cellular adhesion markers ICAM-1 and VCAM-1 were analysed by ELISA. Data analysis was conducted using SPSS 20.00, with significance set at p < 0.005. Results: Manganese (Mn) levels were significantly higher in haemodialysis patients (p = 0.019). Copper (Cu), Molybdenum (Mo), Vanadium (V), Antimony (Sb) and Chromium (Cr) levels were higher in the control group. Zinc (Zn) and Cr levels differed significantly between the control group (p = 0.018; p = 0.007). Cu levels were lower in hypertensive patients (p = 0.011), while Zn and Mn levels were higher in diabetic patients (p = 0.048 and p = 0.004, respectively). Dialysis duration, however, correlated with Sb (r = 0.295; p = 0.01), and Kt/V correlated with Mn, Sb and Cr (r = 0.256, r = 0.272 and r = 0.259, respectively; p = 0.05). Mo levels showed a positive correlation with both pre-dialysis (r = 0.230) and post-dialysis (r = 0.281) creatinine levels, and a negative correlation with post-dialysis GFR (r = −0.294). ICAM-1 and VCAM-1 levels were significantly elevated in dialysis patients (p = 0.001 for both); however, it was not found to be related to variables in the vascular access route. Conclusions: The levels of trace elements and adhesion molecules were examined in haemodialysis patients. High Mn levels indicate a risk of accumulation, while low Cu, Mo, V, Sb and Cr levels may require monitoring for deficiency. ICAM-1 and VCAM-1 levels in haemodialysis patients are associated with some trace elements (Mn and Zn); however, this relationship requires further evidence. In conclusion, the levels of trace elements and adhesion molecules in haemodialysis patients indicate the need for regular monitoring and show that the relationships between creatinine and GFR can be applied to larger patient groups. Full article

Neuroendocrine tumors (NETs) are rare neoplasms arising from the diffuse neuroendocrine system that can range from indolent to highly aggressive diseases. They usually clinically manifest when reaching a significant size or when hepatic metastases develop, leading to overproduction and impaired hepatic metabolism of vasoactive substances. The clinical course of NETs may be complicated by cardiac involvement, known as carcinoid heart disease (CHD), predominantly affecting the right side of the heart. CHD is characterized by specific echocardiographic features, including thickening, reduced excursion and retraction of valvular leaflets, resulting in valvular stenosis or regurgitation. Despite its clinical relevance, awareness of CHD as a complex hormonal sequela of NETs remains limited among cardiologists, and its echocardiographic findings are not universally recognized. This review aims to (a) provide cardiologists with the main principles for understanding CHD pathophysiology; (b) illustrate the main echocardiographic features of CHD, using a stepwise approach; and (c) refine a diagnostic algorithm for detecting cardiac involvement in NET populations and identifying patients at high risk of developing CHD. Full article

Infectious bronchitis virus (IBV) is a highly infectious disease of chickens posing a major threat to poultry production worldwide. Due to genetic and antigenic variability, it is difficult to establish one universal protocol for vaccination against IB. The protectotype vaccination strategy, combining antigenically distinct vaccine strains, offers broader cross-protection than homologous IBV vaccination approaches. This field study aimed to establish serological baseline values in broiler chickens in a protectotype vaccination program using Avishield IB H-120 and Avishield IB GI-13, and to evaluate the program’s efficacy under field conditions on commercial farms where heterologous IBV genotypes were detected. The study was conducted on 25 broiler farms in 2023–2024. Day-old chicks were vaccinated via coarse spray. Blood samples were taken from them at six weeks of age and analyzed using IDEXX and BioChek ELISAs, and cecal tonsils were tested by real-time RT-PCR. Production performance was assessed using the European production efficiency factor (EPEF), with a cut-off value of 360. No IB-related clinical signs were reported, and the mean EPEF of the included farms was 408.7. The detected IBV genotypes were Mass + 793B, 793B, and 793B + VAR2. The presence of heterologous VAR2 did not negatively impact flock health or performance. Preliminary serological baseline values for vaccinated, non-VAR2 farms were established at 891–1332 (in chicks tested with the IDEXX ELISA) and 4193–5204 (in those tested with the BioChek assay). The H-120 + GI-13 program demonstrated high field efficacy, reliable serological profiles, and cross-protection in the presence of heterologous IBV. Continuous monitoring of IBV genotypes remains essential. Full article

This study demonstrates the feasibility of using a T7 phage display platform to deliver a library of tick antigens as a vaccine to disrupt tick feeding in cattle. Cattle were vaccinated at three-week intervals via intradermal and intramuscular routes with a cocktail of male and female Amblyomma americanum T7 phage display cDNA libraries, with and without adjuvant. ELISA and Western blot analyses confirmed that vaccinated cattle mounted immune responses directed against phage-displayed tick proteins rather than the T7 phage backbone. Vaccine-induced antibodies recognized both native tick salivary gland proteins and selected recombinant salivary proteins, indicating effective antigen presentation and biologically relevant immunity with binding to native tick saliva proteins. The adjuvanted formulation elicited significantly stronger immune responses than phage-only immunization. Immunized cattle exhibited robust immune memory, evidenced by a pronounced anamnestic response following tick infestation. This immunity translated into measurable anti-tick effects, including reduced tick feeding efficiency and blood ingestion. Tick reproductive success was severely compromised, with larval hatching declining from 54% in ticks fed on control cattle to 4% in ticks fed on immunized cattle. This study establishes a practical and scalable T7 phage-displayed whole-tick antigen platform capable of inducing durable anti-tick immunity in cattle. Full article

Objectives: This study aimed to determine whether adults with tinnitus exhibit altered phase-specific mandibular kinematics during saliva swallowing and increased prevalence of tongue thrust and Eustachian-tube insufficiency versus tinnitus-free controls. Methods: This was a cross-sectional case–control study including adults with tinnitus and controls. Standardized computerized kinesiography recorded three spontaneous saliva swallows per participant. Primary outcomes were opening/closing time (OCT) and post-closure stabilization time (STT); total swallowing time (SWT) was secondary. Tongue thrust (TT) and tubal insufficiency (TI) were assessed clinically. Distributional assumptions were checked with Shapiro–Wilk; between-group comparisons used two-sided Mann–Whitney U tests and Fisher’s exact tests (TT, TI). Effect sizes included rank-biserial correlation (r), Hodges–Lehmann median difference (Δ), and odds ratios (ORs) with 95% confidence intervals. Co-occurrence of TT and TI and their relationships with OCT, STT, and SWT were evaluated within strata (cases vs. controls) using Fisher’s exact test, φ, Mann–Whitney U tests, and Spearman’s ρ. Given the marked imbalance in age and sex between groups, unadjusted non-parametric comparisons were complemented by multivariable models with adjustment for age and sex. An omnibus non-parametric combination test summarized case–control differences across OCT, STT, and SWT. Results: Statistical analysis was performed on 77 cases with tinnitus and 78 controls. Tinnitus cases showed longer OCT (1.75 ± 0.92 vs. 1.12 ± 0.62 s; p < 0.001; r ≈ 0.40; Δ ≈ +0.60 s) and STT (1.44 ± 0.88 vs. 0.84 ± 0.62 s; p < 0.001; r ≈ 0.42; Δ ≈ +0.60 s), while SWT differed modestly and was not significant (2.75 ± 0.69 vs. 2.57 ± 0.65 s; p = 0.115; r ≈ 0.15; Δ ≈ +0.18 s). TT was more frequent in cases (18.2%) than controls (6.4%; OR = 3.05, 95% CI 1.08–8.61; p = 0.029), whereas TI occurred in 16.9% of cases and 0% of controls (corrected OR = 32.85, 95% CI 1.92–563.49; p < 0.001). Within tinnitus cases, TT and TI did not show meaningful co-occurrence (φ ≈ −0.03; p = 1.00). TT+ tinnitus patients exhibited markedly prolonged OCT compared with TT− (median 2.22 vs. 1.45 s; Δ ≈ +0.88 s; r ≈ 0.60; p < 0.001), whereas STT and SWT were minimally affected; TI was not materially associated with any swallowing-time parameter. Spearman analyses confirmed a moderate monotonic association between TT and OCT in tinnitus cases (ρ ≈ 0.40; p < 0.001), with all other correlations small and clinically negligible. Age- and sex-adjusted analyses confirmed longer OCT and STT in tinnitus cases, whereas SWT remained non-significant; TT and TI also remained more frequent in cases after adjustment. The omnibus test indicated a clear global separation between groups across OCT, STT, and SWT (permutation p < 0.001). Conclusions: Adults with tinnitus exhibit a distinct swallowing signature characterized by prolonged OCT and STT, together with higher prevalence of TT and TI. TT in tinnitus patients is specifically linked to a pronounced prolongation of OCT, while STT and SWT remain largely unchanged, and TI shows no relevant impact on kinematic indices. Full article

Background: Oral iron supplementation or food fortification is essential for managing or preventing iron deficiency but often causes gastrointestinal side effects. While solubility has traditionally been considered a requirement for iron uptake via the DMT1 transporter, recent evidence shows that insoluble iron can also be absorbed through endocytosis, raising questions about particle size and epithelial responses. Methods: Human intestinal cell lines (Hutu-80 and Caco-2) were exposed to physiologically relevant but elevated iron levels (0.5 mM Fe, 48 h) as ferric pyrophosphate, ferrous fumarate (both prone to precipitation), and soluble ferric EDTA. Cell survival and COX-2 protein were quantified by ELISA, solubility by ICP-OES, and particle size in cell culture medium by dynamic light scattering analyses. Results: Ferric pyrophosphate (0.62–3.8 μm) markedly increased COX-2 expression in Hutu-80 cells (254% ± 37%, n = 3, p = 4.11 × 10⁻⁵) and in Caco-2 cells (78% ± 8%, n = 3, p = 0.01) compared to the control. Ferrous fumarate (237–866 nm) also induced COX-2, but only in Hutu-80 cells (62% ± 11%, n = 3, p = 0.04), whereas ferric EDTA showed no effect in either cell line. COX-2 induction was associated with larger particles in the medium (≥237 nm), whereas smaller particles (<146 nm) were not. Conclusions: Particle size appears to be a critical determinant of cell survival and iron-induced epithelial COX-2 expression. Iron compounds that present as both soluble and particulate forms may optimize bioavailability, but controlling aggregate size (<146 nm) could reduce inflammatory signaling. These findings may have important implications for cell culture systems and warrant in vivo validation in iron supplemental studies. Full article

Aquaporins (AQPs) are a family of small integral membrane proteins that mediate the selective transport of water and, in some cases, small solutes such as glycerol and hydrogen peroxide. In the skin, distinct AQP isoforms are expressed throughout the epidermis, dermis, and hypodermis, where they play key roles in maintaining hydration, regulating keratinocyte and fibroblast proliferation, modulating inflammatory responses, and preserving overall tissue integrity. Increasing evidence indicates that aberrant AQP expression or function contributes to skin carcinogenesis, influencing tumor initiation, local invasion, metastasis, and responses to microenvironmental stress. Alterations in specific AQP isoforms have been associated with both major classes of cutaneous malignancies—non-melanoma skin cancers (NMSC), including basal cell carcinoma and squamous cell carcinoma, as well as malignant melanoma (MM)—yet their mechanistic contributions remain incompletely understood. This review synthesizes current knowledge on the involvement of each AQP isoform in skin cancer pathogenesis and progression, integrating findings from molecular, cellular, and in vivo studies. By clarifying the diverse roles of AQPs in cutaneous malignancies, this work aims to support the development of targeted interventions and guide future research in this rapidly evolving field. Full article

Background/Objectives: The Mitrofanoff appendicovesicostomy (MAV) is the gold standard for creating a continent catheterizable channel in children unable to perform clean intermittent catheterization (CIC) through the native urethra. Minimally invasive surgery has progressively replaced open techniques in pediatric urology, offering improved recovery and favorable cosmetic outcomes, and robotic-assisted Mitrofanoff has gained popularity in recent years. However, the high costs and limited availability of robotic systems create disparities in access to pediatric urologic reconstruction, particularly in low- and middle-income countries. In this context, the laparoscopic Mitrofanoff (MAV-L) and the laparoscopic-assisted Mitrofanoff (MAV-LA) represent practical, cost-effective alternatives, valuable in institutions without robotic platforms or in resource-limited settings. Recent evidence demonstrates that advanced laparoscopic approaches remain feasible even for complex urological procedures, supporting their continued relevance in the robotic era. Methods: We conducted a retrospective case series including seven male children (aged 9–12 years) who underwent MAV between 2018 and 2023. Peri-operative data included demographics, operative time, length of hospitalization, and complications. Functional and aesthetic outcomes were assessed during long-term follow-up. Statistical analysis accounted for the small sample size by using non-parametric tests where appropriate. Results: Three patients (43%) underwent MAV-L and four (57%) MAV-LA. Mean operative time appeared longer in MAV-L (273.3 ± 20.5 min) than in MAV-LA (203.8 ± 24.3 min; exploratory p = 0.019). Hospital stay was 9 ± 0.8 days vs. 7.5 ± 0.5 days (p = 0.026). During follow-up (43.3 ± 10.9 vs. 26.3 ± 5.4 months; p = 0.034), two complications occurred, both in the MAV-L group (stomal stenosis and channel leakage). All patients reported excellent continence, ease of catheterization, and high cosmetic satisfaction. Conclusions: Both laparoscopic and laparoscopic-assisted Mitrofanoff techniques are safe, feasible, and effective in children. Favorable cosmetic satisfaction was reported in the fully laparoscopic subgroup, based on subjective assessment. Importantly, these laparoscopic techniques are sustainable alternatives to robotic surgery, offering accessibility and high-quality reconstructive care even in centers with limited financial and technological resources. Full article

Background: Recent advances in multimodal therapies have increased the potential for resectability of borderline resectable and locally advanced pancreatic ductal adenocarcinoma (PDAC). We herein describe the conservative resection strategy adopted at our institution and the oncological outcomes of patients with PDAC and arterial involvement. Methods: This retrospective single-centre study included patients diagnosed with PDAC and radiologic evidence of arterial involvement who underwent surgical exploration between January 2014 and June 2024. All patients received induction chemotherapy (±radiotherapy). Survival outcomes were analyzed using the Kaplan–Meier and Cox proportional hazards models. Logistic regression analyses were used to identify predictors of resectability and recurrence. Results: A total of 76 patients were included: 59 underwent pancreatic resection with arterial divestment (AD) in case of persistent arterial involvement and 17 were deemed unresectable at laparotomy. Neoadjuvant folfirinox was significantly associated with increased odds of resection (HR = 3.23, 95% CI: 1.59–9.90, p = 0.040). Median overall survival from diagnosis was 33 months (29–39) in resected patients and 26 months (16–29) in non-resected patients (p = 0.0176). Surgical resection and Ca 19,9 normalization after induction therapy were associated with reduced mortality risk (HR = 0.38, 95% CI: 0.19–0.75, p = 0.005 and HR = 0.56, 95% CI: 0.35–0.88, p = 0.014, respectively). Conclusions: Despite a limited sample size and retrospective nature, these findings highlight the value of multimodal strategies in managing PDAC with arterial involvement. AD represents a valuable technique associated with acceptable outcomes in selected patients. Future interventional prospective studies are needed to optimize patient selection and validate the prognostic role of extended surgical procedures. Full article

Background: Measles remains a global public health threat despite the availability of an effective vaccine and substantial progress toward elimination in many countries. Outbreaks in highly vaccinated settings suggest that waning vaccine-induced immunity, particularly among adults, may create silent susceptibility gaps capable of sustaining viral transmission. Aim: To evaluate age- and cohort-specific patterns of measles-specific antibody levels in Mexican adults and to examine evidence suggesting lower vaccine-induced antibody levels in younger vaccinated cohorts. Methods: A cross-sectional seroepidemiological study was conducted among 302 voluntary blood donors aged 18–70 years. Demographic, clinical, and vaccination data were collected through structured interviews. Serum anti-measles-virus (MV) IgG levels were quantified using a commercial ELISA. Antibody concentrations were analyzed according to age, sex, vaccination history, self-reported measles infection, and historical vaccination strategies. Multivariate linear models were applied to identify factors associated with IgG levels. Results: Anti-MV IgG seropositivity (>200 mIU/mL) was 67.2%, with a geometric mean concentration (GMC) of 270.43 mIU/mL. A positive correlation was observed between age and antibody levels (r_s = 0.161, p = 0.005). Individuals born before the introduction of the measles vaccine (pre-1970) had significantly higher GMCs (1096.63 mIU/mL) than younger cohorts. A history of natural infection tended to be associated with higher antibody levels (GMC: 428.38 vs. 257.24 mIU/mL; p = 0.051). In multivariate analysis, historical vaccination strategy emerged as the primary factor associated with antibody levels, whereas age alone was not significant. Conclusions: Cohort-specific differences in measles IgG levels suggest generational patterns of immunity and are consistent with diminished vaccine-induced antibody levels in younger adults in the absence of natural boosting. These findings highlight the importance of ongoing serological surveillance in post-elimination settings and underscore the need for targeted public health interventions. Full article

Maize is highly vulnerable to aflatoxin contamination, constituting a serious food safety and public health concern. This study explored the relationship of extreme flood events in September 2020 (Storm Ianos) and September 2023 (Storm Daniel) in the Thessaly region, Greece, which occurred post-harvest, and aflatoxin contamination patterns in maize harvests of both the flood years and the following years (2021 and 2024). A total of 573 maize samples collected between 2019 and 2024 were analyzed using an enzyme-linked immunosorbent assay (ELISA). A 10 μg/kg cutoff was used (the ELISA upper limit of quantification); results > 10 ppb were classified as elevated concentrations. Overall, aflatoxins were detected in 47.8% of samples, with 14.0% having concentrations exceeding 10 μg/kg. The 2021 harvest year exhibited an exceedance rate of 28.75% (23/80), while the 2024 harvest showed a rate of 14.68%, 37/252. Exceedance rates during flood years were comparatively low; however, the maize harvested in the years following the flood events demonstrated a two-fold increase in the detection rate (60.2% vs. 30.7%) and a significant increase in exceedance percentages (18.07% vs. 8.30%) relative to non-flood years in the preceding period. While drought and heat remain the primary field risk factors for aflatoxin production, the correlational findings suggest that extreme floods may indirectly influence aflatoxin risk by increasing kernel damage, prolonging grain moisture, and disrupting post-harvest handling and storage, underscoring the need for continuous monitoring and robust mitigation strategies in flood-prone agricultural regions. Full article

Background/Objectives: Meloxicam (MLX) is a nonsteroidal anti-inflammatory drug (NSAID) recommended for treating acute and chronic pain in dogs, frequently administered prophylactically to mitigate postoperative pain; however, its utility is limited by characteristic NSAID-associated adverse effects, such as gastrointestinal side effects. Nanosystems offer the potential to minimize adverse effects by sustaining drug release. Therefore, this study assessed the pharmacokinetics of MLX nanoencapsulation in female dogs undergoing ovariohysterectomy using a population pharmacokinetic (PopPK) modeling approach. Methods: MLX-loaded polymeric nanocapsules (NC-MLX) were prepared using the nanoprecipitation method and characterized by zeta potential, pH, mean diameter, particle size distribution, and drug content. Dogs received 0.2 mg/kg of either NC-MLX or free MLX orally, 4 h before surgery, and plasma samples were analyzed using an HPLC-PDA method. Pharmacokinetics were characterized by non-compartmental analysis and PopPK modeling. Several compartmental structures, variability models, and residual error models were explored, and relevant covariates were investigated. Results: NC-MLX had an average diameter of 326 ± 13 nm, a zeta potential of −26.2 ± 6.4 mV, and drug loading of 99.47% ± 0.01%. NC-MLX showed a significant increase in the t_1/2 (36.99 ± 17.26 h) of MLX compared to the free drug (15.22 ± 4.4 h). The best-fitting PopPK model was a two-compartment model with double extravascular first-order absorption rate constants (Ka₁ and Ka₂), including a lag time for Ka₂ and linear elimination, describing the second peak observed in several animals. The nanoformulation was a significant covariate for Tlag₂, delaying the time for absorption (1.22 and 2.55 h for free MLX and NC-MLX, respectively) and increasing V₂ (0.134 and 0.402 L/kg for free MLX and NC-MLX, respectively). External model validation showed that the final PopPK model accurately predicted plasma concentrations, with MPE% and RMSE values below 15%. Conclusions: Our findings suggest that NC-MLX alters MLX absorption and distribution profiles, supporting its potential as an alternative for postoperative pain management in dogs. Full article