Biology

In silico analyses, including absorption, distribution, metabolism, excretion, and toxicity (ADMET) profiling and drug-likeness evaluations, constitute powerful tools in modern drug discovery. These computational methods enable high-throughput screening of molecular libraries to assess therapeutic potential while identifying unfavourable pharmacokinetic and toxicological profiles early in the development process. To bridge the gap between computational models and empirical data, this study evaluates the reliability and limitations of predictive ADMET profiling by comparing in silico outputs across multiple platforms and referring them to available in vitro experimental data. The empirical dataset comprises recently published in vitro assays focusing on inhibitors of the protein tyrosine phosphatase 1B (PTP1B) enzyme. ADMET predictions were generated using three widely utilized, non-commercial platforms: SwissADME, ADMETlab 3.0, and admetSAR 3.0. The comparative analysis encompassed physicochemical parameters (MW, logP, TPSA), medicinal chemistry alerts (PAINS, ALARM NMR), absorption metrics (Caco-2, HIA), and multiple toxicity endpoints (Ames mutagenicity, DILI, hERG inhibition, nuclear receptor signaling, and mitochondrial toxicity). Significant differences observed in toxicity predictions across the platforms highlight the critical necessity of employing multi-tool predictive strategies to obtain a reliable and comprehensive outlook.

The Japanese eel (Anguilla japonica) is an economically important aquaculture species, and its reproductive development and artificial propagation have received considerable attention. However, the absence of a stable in vitro female germ cell culture system has hindered investigations into oogenesis, reproductive regulation, and germplasm conservation. In this study, cells isolated from immature Japanese eel ovaries were enriched through differential adhesion, Percoll density-gradient centrifugation, and single-cell cloning, followed by optimization of culture conditions. A stable oogonial stem cell-like cell line of A. japonica, designated AjOSCL, was established and characterized via morphological observation, proliferation assays, colony-forming assays, karyotyping, immunofluorescence staining, alkaline phosphatase detection, and transcriptomic sequencing. AjOSCL maintained stable growth and self-renewal capacity during continuous culture for over 35 passages in vitro, with a cumulative culture duration exceeding 12 months, and retained a predominant chromosome number of 2n = 38 (consistent with the chromosome number of A. japonica), suggesting chromosomal stability during long-term culture. AjOSCL exhibited strong alkaline phosphatase activity and expressed germline-related markers (e.g., Vasa and Nanos2), together with pluripotency-related genes or proteins such as Nanog and Oct4. Transcriptomic analysis further revealed that AjOSCL displayed a molecular profile compatible with a germline stem cell-like phenotype, with enrichment of pathways related to germ cell maintenance and oocyte development, including ECM–receptor interaction, TGF-β signaling, Wnt signaling, and oocyte meiosis. Collectively, these results indicate that AjOSCL may serve as a renewable in vitro model for studying oogonial maintenance, offering a foundation for future investigations into female germ cell biology and germplasm conservation in A. japonica.

Current preclinical studies evaluating cGAS inhibition as a novel anti-inflammatory strategy rely predominantly on young murine models, leaving the impact of aging largely unexplored. At baseline, aged (56-week-old) female mice, in both WT and cGAS^-/- mice, demonstrated more pronounced oxidative stress (malondialdehyde) and diminished antioxidant capacity (glutathione) relative to young (8-week-old) controls. Upon the induction of liver ischemia–reperfusion injury (IRI), the most severe hepatic damage was provoked exclusively in aged cGAS^-/- mice. This exacerbated injury was confirmed by elevated liver enzymes, higher histological scores, widespread apoptosis (activated caspase-3), enhanced macrophage accumulation (F4/80⁺ cells), and increased serum cell-free DNA as a tissue damage marker. Additionally, aged cGAS^-/- mice exhibited a significant surge in serum IL-6 and gut barrier dysfunction (FITC-dextran assay) at 6 h post-injury. Mechanistically, while cGAS^-/- macrophages showed blunted inflammatory signaling upon lipopolysaccharide (LPS) stimulation, co-incubation with D-galactose, which induces intracellular peroxide accumulation, restored their pro-inflammatory responses to WT levels. Concurrently, starvation protocols revealed that cGAS^-/- hepatocytes were more vulnerable to mitochondrial oxidative stress (MitoSOX) than WT cells. Ultimately, aging critically modifies the cGAS-deficient phenotype, warranting the careful re-evaluation of cGAS therapeutic models in aged populations.