Exogenous glucagon-like peptide 2 (GLP-2) administration increases small intestinal growth and mesenteric blood flow, although the vascular mechanisms involved remain unclear. The objective of this study was to characterize the effects of exogenous GLP-2 administration on glucagon-like peptide 2 (GLP-2)-, vasoactive intestinal peptide (VIP)-, serotonin (5-HT)-, and acetylcholine (ACh)-mediated vascular relaxation in isolated bovine mesenteric arteries (MA) and mesenteric veins (MV). Ten steers received repeated subcutaneous injections of bovine serum albumin or GLP-2 for 48 h before vessel collection. Vascular dimensions, vasoactive responses using myography, and mRNA expression of receptors associated with vasorelaxation were evaluated. Exogenous GLP-2 did not directly induce vasorelaxation in isolated MA or MV despite detectable GLP2R mRNA expression in both vessels, suggesting that GLP-2-mediated increases in mesenteric blood flow occur through indirect mechanisms. Vasoactive intestinal peptide induced dose-dependent vasorelaxation in both vessel types, and only VIPR2 mRNA was detected. Serotonin-mediated responses were not altered by exogenous GLP-2 treatment. In contrast, exogenous GLP-2 enhanced ACh-mediated vasorelaxation in the MA at greater agonist concentrations and increased NOS3 mRNA expression in the MV. These findings suggest that cholinergic signaling pathways may contribute to GLP-2-associated vasoregulation in the bovine MA.

Advanced systemic mastocytosis (AdvSM) is driven by KIT D816V in approximately 95% of cases. Accurate assessment and prediction of clinical course and disease outcome before and during treatment with the KIT inhibitors midostaurin and avapritinib may facilitate optimization of treatment strategies. We retrospectively evaluated clinical, morphologic, laboratory, and genetic predictors of outcome in 79 midostaurin- and 176 avapritinib-treated patients. The number of mutations at baseline in SRSF2/ASXL1/EZH2 (midostaurin) and in SRSF2/RUNX1/SETBP1 (avapritinib) delineated highly predictive 3-tier risk classification systems. Treatment with avapritinib was associated with an early response of the KIT D816V variant allele frequency; a ≥10% reduction in peripheral blood at day 15 indicated improved overall survival (OS). Within 3 months of starting treatment, a ≥25% increase in alkaline phosphatase (AP) levels (AP flare) was observed in 46/47% of midostaurin/avapritinib-treated patients and was associated with longer OS. Multivariable analyses identified a ≥25% reduction in the KIT D816V allele burden (midostaurin), normalization of serum albumin levels (avapritinib), and normalization of hemoglobin and/or platelet levels (midostaurin and avapritinib) within 6 months as significant predictors for prolonged OS. In conclusion, genetic profiling, dynamic laboratory parameters, and KIT D816V monitoring improve outcome prediction in patients with AdvSM on midostaurin or avapritinib.

As metabolomics data are typically acquired across multiple batches over months or even years, the integrated metabolite data sets often face complex missing-value challenges arising from fluctuations in instrument performance, variation in detection windows, and heterogeneity among analytical platforms. Accurate imputation of missing values is therefore a crucial strategy to addressing these challenges and enhancing the performance of downstream metabolomics analyses. In this study, we utilized large-scale authentic plasma and serum metabolite data sets (n = ∼3,000 metabolites) in Holstein cattle (n = ∼3,000 individuals), with each data set consisting of 2 batches, to evaluate the performance of 5 imputation methods (MIRTH, PIXANT, EM, KNN, and RF) to impute missing metabolites within batches and non-overlapping metabolites between batches. We demonstrated that RF yielded the best imputation performance in most cases, with accuracies reaching approximately 0.6 ∼0.7 for within-batch imputation and approximately 0.3 ∼0.4 for cross-batch imputation. Subsequently, we assessed the effect of imputation accuracy on the performance of metabolite-trait association analysis for 5 milk production traits using RF-imputed metabolite data. Compared with significantly associated metabolites identified using the original metabolite data, analysis using the imputed metabolite data resulted in substantial reductions in statistical power as well as increases in false-positive associations, particularly when imputed data from different batches were merged. Further efforts are needed to improve the accuracy for cross-batch imputation by developing new imputation methods and increasing the number of overlapping metabolites across batches.

The study aimed to examine the physical properties of milk replacer (MR) supplemented with linseed oil or ethyl esters of polyunsaturated fatty acids of linseed oil, and the subsequent impacts on nutrient and fatty acid (FA) digestibility, growth performance, health status, and some immune response and oxidative stress parameters in pre-weaned dairy calves. Eighteen Holstein calves were assigned to one of 3 treatments: a control MR with no supplemental fat added (CON), MR supplemented with 30 mL/d of linseed oil (LO), or MR supplemented with 30 mL/d of ethyl esters of polyunsaturated fatty acids of linseed oil (EELO). Physical properties of MR, including creaming index (CI), droplet size distribution, and microstructure, were evaluated after mixing/homogenizing with water. Apparent total-tract digestibility of nutrients was determined by fecal collection during 2 periods (22-25 and 50-53 d of age). Digestibility was calculated for dry matter, organic matter, crude protein, crude fat, and individual fatty acids (C16:0, C18:0, C18:1, C18:2, and C18:3). Titanium dioxide (TiO2) was included in the diet as an indigestible marker. Health parameters were recorded daily from 12 to 53 d of age, and some immune and oxidative stress indicators were measured in blood samples collected on 12, 25, 43, and 53 d of age. Performance parameters such as MR and starter mixture intake, total DMI, ADG, feed efficiency as well as nutrient intakes, nutrient digestibility coefficients, immune status, or oxidative stress parameters were analyzed as repeated measures using a linear model and the GLIMMIX procedure in SAS. Supplementation of MR with LO or EELO affected CI and droplet size characteristics, with EELO showing improved MR stability compared with LO, including a lower CI, more favorable droplet size distribution, and reduced fat globule coalescence. Fat supplementation did not affect DMI. However, final BW, ADG, and feed efficiency tended to be greater in calves receiving EEPUFALO. Digestibility of OM and CP was greater in the EELO group in both periods, whereas LO supplementation reduced C18:2 digestibility during the first experimental period. Digestibility of selected FA, including C16:0 and C18:3, was higher in calves supplemented with EELO. Irrespective of treatment, older calves exhibited greater digestibility of crude fat and PUFA, indicating an age-related improvement in lipid digestion. Fat supplementation reduced the number of days with diarrhea and the number of treatment days, with the most pronounced effects observed in the EELO group. Supplementation with EELO was associated with reduced concentrations of measured inflammatory markers, particularly TNF-α, and a tendency toward lower serum haptoglobin. In addition, EELO supplementation improved oxidative status, reflected by reduced total oxidant status, increased total antioxidant status, and a reduced oxidative stress index. In conclusion, EELO exhibited superior physical, nutritional, and biological effects compared with LO, demonstrating that the chemical form of FA is a critical factor influencing their digestibility, bioavailability, and health-promoting effects in pre-weaned dairy calves.

Apical periodontitis (AP) is characterized by persistent inflammation and progressive periapical bone resorption, processes increasingly associated with NLRP3 inflammasome activation and cGAS-STING-related inflammatory signaling. However, pharmacological approaches targeting these inflammatory pathways and their upstream regulatory mechanisms remain limited. Here, we investigated whether liquiritigenin, a natural flavonoid derived from licorice, attenuates inflammatory injury and periapical bone loss through TRIM31-associated regulation of NLRP3 and modulation of cGAS-STING-related signaling. Structure-based virtual screening identified several natural compounds with predicted TRIM31-binding potential, among which liquiritigenin showed the strongest protective activity against LPS/ATP-induced loss of cell viability. Pronase-protection and cellular thermal shift assays showed that liquiritigenin increased TRIM31 stability, supporting potential intracellular target engagement without establishing direct physical binding. LPS/ATP stimulation reduced TRIM31 abundance and increased NLRP3 expression, IL-1β release, cleaved caspase-1, mature IL-1β, and cleaved GSDMD. Liquiritigenin reversed these changes, whereas TRIM31 knockdown partially weakened its effects. Moreover, liquiritigenin preferentially enhanced K48-linked, but not K63-linked, NLRP3 ubiquitination; graded TRIM31 knockdown progressively reduced this response. CHX-chase and MG132 experiments further supported accelerated, proteasome-associated turnover of NLRP3. Liquiritigenin also reduced cGAMP production, STING phosphorylation, downstream TBK1-IRF3 phosphorylation, and the expression of IFN-β, ISG15, and CXCL10 under LPS/ATP stimulation. In C57BL/6 mice with LPS/ATP-induced periapical inflammatory bone loss, liquiritigenin alleviated periapical bone loss, improved micro-CT parameters, reduced inflammatory infiltration, restored TRIM31 expression, and attenuated NLRP3 accumulation and TBK1-IRF3 signaling without significantly elevating serum ALT or AST levels. Collectively, these findings identify liquiritigenin as a candidate natural compound for limiting inflammatory periapical bone loss. Its protective effects are partly associated with TRIM31-mediated K48-linked ubiquitination and proteasome-associated turnover of NLRP3 and are accompanied by attenuation of cGAS-STING-related signaling.

Polylactic acid (PLA) is currently one of the most widely used bio-based plastics, posing exposure risks to humans through various environmental pathways. Therefore, this study evaluated the effects of PLA nanoparticles (NPs) on the testes and the potential mechanisms involved. C57BL/6 J mice were administered PLA NPs (0.1, 1, and 10 mg/kg/day) for 4 weeks. Additionally, supplementary validation was conducted using transient receptor potential cation channel subfamily m member 2 (TRPM2) knockout models and MitoTEMPO-treated models. PLA NPs induced marked histological damage to the testes, resulting in impaired spermatogenesis and suppressed hormone synthesis. PLA NPs elevated reactive oxygen species levels, exacerbated oxidative stress, and mediated mitochondrial damage. Furthermore, PLA NPs activated TRPM2 and induced calcium ion dysregulation, leading to a significant increase in intracellular calcium levels. The study further revealed that PLA NPs markedly elevated serum IL-1β levels and activated the inflammasome pathway, thereby inducing pyroptosis in mouse testicular cells and resulting in reproductive toxicity. Collectively, these findings indicated that PLA NPs activate the TRPM2 pathway through oxidative stress-induced mitochondrial damage, subsequently inducing calcium dysregulation and causing pyroptosis in mouse testes. Our research provides new insights and evidence for the reproductive toxicity assessment of PLA NPs.

Chronic Kidney Disease is a global health challenge and its early diagnosis is still a challenge, particularly when relying on conventional biomarkers like serum creatinine. In this systematic review, PRISMA guidelines were followed with a comprehensive search on various databases focusing on studies assessing estimated cystatin C levels, and creatinine levels among patients with chronic kidney disease, type 2 diabetes mellitus and hypertension. The Joanna Briggs Institute's critical assessment tool was used to estimate quality and risk bias of included studies. This systematic review included 21 studies with maximum studies demonstrating cystatin C based eGFR may provide more reliable and accurate estimate of kidney function than creatinine based eGFR, specifically for early identification of kidney dysfunction. The findings suggest that serum cystatin C is an early diagnostic and prognostic marker and its incorporation into routine testing will increase the clinical utility by improving risk stratification. In general, this systematic review highlights the diagnostic and prognostic utility of serum cystatin C for early detection of kidney dysfunction.

The adrenocorticotropic hormone stimulation test (ACTHST) is widely used to evaluate adrenal reserve in dogs. This study assessed the effects of refrigerated storage of tetracosactide (Cosacthen) after opening for up to three weeks and investigated hematological, biochemical, and electrolyte changes following ACTH stimulation. Ten healthy beagle dogs (five males and five females) were enrolled. Each dog received tetracosactide (5 μg/kg, IV) at vial opening (T0) and at 7-day intervals over 3 weeks (T1 to T3), using aliquots from the same vial stored at 2-8°C. Blood samples were collected before and 1 hour after ACTH administration to measure cortisol, hematology, biochemistry, and electrolytes. Clinical monitoring was performed after each test. Cortisol concentrations increased significantly after ACTH administration at all time points (T0-T3). Delta-cortisol was higher at T1 than at T0 (P = 0.0104) and lower at T2 than at T1 (P = 0.0059), with no differences among T0, T2, and T3. After ACTH administration, red blood cell count, hematocrit, hemoglobin, albumin, alkaline phosphatase, triglycerides, cholesterol, potassium, and total calcium decreased significantly, whereas glucose increased. All values remained within reference intervals, and no adverse effects were observed. Refrigerated storage of Cosacthen after opening did not alter cortisol responses. Post-ACTH changes in laboratory variables should be considered when interpreting results.

Nanotechnology-driven feed additives provide an effective approach to enhance aquaculture sustainability through the improved delivery and bioavailability of bioactive compounds. This study aimed to formulate eugenol-loaded sodium alginate beads (E-SABs) as a biopolymer-mediated sustained-delivery system and to evaluate their effects on growth performance, antioxidant and immune status, and resistance to Aeromonas hydrophila infection in Nile tilapia (Oreochromis niloticus). The formulated E-SABs had a mean particle (Z-average) diameter of 79.66 nm, a polydispersity index of 0.413 and a zeta potential of -9.18 mV, and released eugenol in a sustained manner in vitro (20-25% burst release within 2 h, reaching 96% cumulative release at 24 h; Korsmeyer-Peppas model, R2 = 0.999, release exponent n = 0.69, indicating non-Fickian diffusion). Juvenile Nile tilapia (initial weight approximately 20-100 g; n = 100) were randomly allotted to four dietary groups (a basal-diet control and E-SABs at 0.25, 0.50 and 1.0 mg kg-1 feed) and fed for 30 days. Final body weight, specific growth rate and survival were significantly higher, and feed conversion ratio significantly lower, in all E-SAB groups than in the control (P < 0.05), with the greatest improvement at 1.0 mg kg-1 (e.g., final weight 69.90 ± 0.45 g vs. 59.20 ± 0.52 g; survival 98.8 ± 0.85% vs. 90.0 ± 1.5% in the control). E-SAB-fed fish also showed higher serum total protein, albumin and globulin, and lower AST, ALT, ALP, cholesterol and glucose, than the control (P < 0.05). Innate immunity was enhanced, with dose-dependent increases in phagocytic activity and phagocytic index, lysozyme activity and serum IgM (P < 0.05), together with higher activities of the antioxidant enzymes SOD, CAT and GPx and lower malondialdehyde levels (P < 0.05). These physiological changes were paralleled by up-regulation of genes governing somatic growth (GH, IGF-1 and IGF1-R), immune signalling (IL-1β and IL-8) and antioxidant defence (SOD, CAT and GPx). Following experimental A. hydrophila challenge, cumulative survival at day 15 was significantly higher in E-SAB-fed fish (60-85%, dose-dependent) than in the control (30%) (P < 0.05). Collectively, these findings establish alginate-eugenol microbeads as an effective biomacromolecular delivery platform that couples sustained release with immunomodulation, and support the dietary inclusion of E-SABs, particularly at 1.0 mg kg-1 feed, as a practical, antibiotic-free strategy for improving growth, immune competence and disease resilience in Nile tilapia aquaculture; further evaluation under commercial farm conditions is recommended.