Our Goal is to sensitize human colorectal cancer (CRC) to the differentiation-inducing effects of retinoic acid (RA) agents by suppressing WNT signaling. Previously, we reported that the WNT and RA pathways are functionally linked, and in CRC, APC mutation generates a WNT:RA imbalance leading to incomplete differentiation and stem cell (SC) overpopulation. Hypothesis: Therapeutically inhibiting WNT signaling enhances the ability of retinoid agents to induce growth suppression and differentiation. We tested drug combinations of: (1) anti-WNT agents (Sulindac & Piroxicam) that produce anti-tumor activity in humans and animals harboring germline APC mutations, with (2) RA agents (All-trans-RA, 9-cis RA) and RA metabolism blocking agent (CYP26A1 inhibitors Liarozole and Talarozole), which inhibit proliferation and induce differentiation of CRC cell lines. Results: The combined effect of anti-WNT agents and RA drugs shows additive or synergistic responses in CRC cell lines compared to single agents. Drug combinations decrease the proportion of aldehyde dehydrogenase-positive SCs and increase neuroendocrine cell differentiation in CRC cells that show a synergistic response to treatment. NanoString profiling shows that therapeutically decreasing WNT signaling in CRC cells significantly changes the effect of RA signaling on gene expression. Treating human CRC organoid cultures and PDX mouse models with WNT:RA-based drug combinations led to greater anti-tumor activity as compared to individual agents. Conclusion: Inhibiting WNT signaling effectively sensitizes CRC cells to the anti-tumor effects of retinoids. These findings have important clinical significance for designing new drug combinations against CRC.

The recent clinical introduction of the piroxicam (PIR) and venlafaxine hydrochloride (VEN) combination has highlighted the need for analytical methodologies capable of simultaneously quantifying both drugs while minimizing the environmental burden of routine pharmaceutical analysis. To date, only a chromatographic method has been reported for this purpose, which is associated with relatively high organic solvent consumption, greater instrumental complexity, and increased energy requirements. A green UV spectrophotometric platform integrated with chemometric modeling was therefore established for the concurrent determination of PIR and VEN in pharmaceutical formulations and as a proof-of-concept application to spiked human plasma. Spectral measurements were performed using a water-ethanol (1:1, v/v) solvent system, whereas methanol was employed solely for plasma protein precipitation. Experimental design efficiency was enhanced by combining Brereton's multilevel calibration design with a Maximin Distance Design (MMD) for validation sample selection, enabling representative calibration and validation datasets to be generated with a reduced number of experimental mixtures. Severe spectral overlap between the two analytes was effectively resolved using six well-established chemometric approaches: CLS, PCR, PLS, GA-PLS, FA-PLS, and MCR-ALS. Among the investigated models, MCR-ALS showed the best overall numerical performance, achieving correlation coefficients of up to 0.9998 for pharmaceutical formulations and 0.9996 for spiked human plasma. The corresponding limits of detection were 0.125 and 0.029 µg mL[-1] for VEN and PIR, respectively, in pharmaceutical formulations, and 0.245 and 0.058 µg mL[-1] in spiked human plasma. Across the investigated concentration ranges, the proposed method also exhibited satisfactory accuracy and precision. The sustainability performance of the analytical platform was evaluated using multiple complementary metrics, with the pharmaceutical assay compared with the previously reported chromatographic procedure according to the evaluated criteria. The proposed pharmaceutical spectrophotometric assay showed a more favorable sustainability profile according to the evaluated criteria, including environmental and practical applicability dimensions. The application to spiked human plasma provided a proof-of-concept assessment of matrix applicability. Nevertheless, additional investigations involving incurred clinical specimens and full bioanalytical validation are required before the methodology can be considered suitable for routine clinical practice or pharmacokinetic studies.

Molecular-targeted therapies require biomarker-driven evaluation to optimize precision oncology strategies; however, translational models integrating clinical outcomes with mechanistic biomarkers are limited. Canine urothelial carcinoma shares molecular features with human muscle-invasive bladder cancer, including activation of VEGFR signaling, providing a valuable comparative oncology platform. Sorafenib is a multi-kinase inhibitor targeting VEGFRs, PDGFRβ, BRAF, and related pathways, but its therapeutic mechanisms and clinical benefit in urothelial carcinoma remain incompletely understood. In this study, 43 dogs with naturally occurring urothelial carcinoma received first-line sorafenib and piroxicam therapy. The overall response rate was 62.8%, with median progression-free and overall survival of 175 (range, 31-665) and 407 (range, 103-1,723) days, respectively. The results exceeded outcomes in historical controls treated with piroxicam alone. Treatment was generally well tolerated, with only three grade 3 adverse events and no grade 4-5 toxicities. Biomarker analyses revealed that treatment-induced hypertension and high tumor CX3CL1 mRNA expression were independently associated with improved overall survival, suggesting that effective VEGFR pathway inhibition and modulation of the tumor immune microenvironment contribute to clinical benefit. These findings support sorafenib-based therapy as a promising strategy for canine urothelial carcinoma and highlight the translational potential of biomarker-driven VEGFR-targeted approaches in precision oncology.

Inflammatory bowel disease (IBD) is a chronic inflammatory disorder of the digestive tract affecting over 10 million individuals globally. While substantial research has focused on the immunologic mechanisms and consequences underlying IBD, less is understood about how mucosal inflammation contributes to metabolic dysregulation, including weight loss and reduced appetite. Here, the authors describe a mouse model of piroxicam-accelerated enterocolitis in interleukin-10-knockout (IL-10-KO) mice to study inflammation-induced metabolic dysregulation. IL-10-KO mice are known to develop spontaneous enterocolitis and have heightened susceptibility to enterocolitis triggered by infections or drugs. However, vivarium conditions and strain background have been reported as confounders in colitis development in this model. Piroxicam, a non-steroidal anti-inflammatory drug (NSAID), has been demonstrated to trigger or accelerate enterocolitis in animal models by increasing mucosal exposure to luminal bacteria. In this protocol, male and female IL-10-KO mice were fed a piroxicam-fortified diet in place of a regular chow diet. Food intake and body weight were measured daily to reflect whole-body metabolic alterations, along with clinical manifestations of enterocolitis such as diarrhea and rectal bleeding. The protocol is efficient and reproducible, inducing enterocolitis simultaneously in multiple mice, and allows assessment of metabolic dysregulation in an inflammatory bowel disease model. Further studies using this protocol may investigate the effects of enterocolitis on other components of metabolic dysregulation, such as energy expenditure and body composition, revealing broader connections between inflammatory bowel disease and host metabolism.

Rheumatoid arthritis (RA) is a chronic immune-mediated disease in which persistent inflammation and oxidative stress contribute to progressive joint damage. This study evaluated the prophylactic effects of sabinene in complete Freund's adjuvant (CFA)-induced arthritis and explored potential molecular associations using integrated in vivo and computational approaches. Rats were allocated to vehicle control, arthritic control, piroxicam, or sabinene groups (15, 30, or 60 mg/kg; n = 6/group); sabinene was administered orally beginning 30 minutes before CFA injection and continued for 28 days. Paw swelling, hematological and biochemical parameters, inflammatory and oxidative-stress markers, and gene expression were assessed, followed by network pharmacology, enrichment analysis, and molecular docking. Sabinene dose-dependently attenuated CFA-induced paw swelling and body-weight loss and improved hematological, hepatic, renal, and inflammatory parameters. Treatment also enhanced antioxidant defenses and reduced lipid peroxidation, prostaglandin E2, 5-lipoxygenase, and anti-cyclic citrullinated peptide antibody levels. Sabinene reduced messenger RNA expression of nuclear factor kappa B (NF-κB), toll-like receptor 4 (TLR4), NOD-like receptor family pyrin domain-containing 3 (NLRP3), caspase-1, gasdermin D, and other pro-inflammatory genes while increasing interleukin-4 and interleukin-10 expression. Network pharmacology identified 133 overlapping sabinene-RA targets, and molecular docking predicted interactions with tumor necrosis factor alpha (TNF-α), interleukin-6 (IL-6), signal transducer and activator of transcription 3 (STAT3), and interferon gamma (IFN-γ). These findings support a prophylactic protective effect of sabinene against CFA-induced arthritis associated with reduced inflammation, improved antioxidant status, and modulation of inflammasome-related gene expression. Further protein-level, functional, and post-induction studies are required to confirm the proposed mechanisms and therapeutic relevance.

The pathology of diabetic foot ulcer (DFU) is characterized by keratinocyte dysfunction, non-resolving inflammation, and oxidative stress. We aim to investigate the effects and mechanisms of piroxicam on DFU healing through regulating mitochondrial function and suppressing inflammation.