Methods: Rats were fed a high-fat diet for 4 weeks to induce obesity, followed by a 4-week treatment period. Thirty rats were allocated into five groups (six rats per group): control (dist. water, orally), HFD control (dist. water, orally), HFD + chrysin (500 mg/kg, orally), HFD + chitosan-NP (60 mg/kg, orally), and HFD + Chrysin-CSNPS (60 mg/kg, orally).

Results: In silico studies revealed that chrysin has a binding energy value of −8.8 kcal/mol to fat mass and obesity-associated (FTO) protein. Also, Chrysin is identified as an inhibitor of several cytochrome P450 enzymes, specifically CYP1A2, CYP2D6, and CYP3A4. Albumin, high-density lipoprotein cholesterol, glutathione, and nitric oxide levels rose, whereas glucose, alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, creatinine, urea, total cholesterol, triglycerides, malondialdehyde, and nitric oxide levels fell upon Chrysin-CSNPS treatment. The histological examination revealed a significant enhancement in the structures of the liver and kidneys.

Discussion: These findings suggest that chrysin could potentially inhibit FTO activity, thereby contributing to a reduction in obesity-related phenotypes. The compound that satisfied Lipinski’s criteria was selected for toxicity prediction.

Conclusion: Chrysin-CSNPS have hypolipidemic properties and an antioxidant role, reducing HFD consequences in the liver and kidney.

Methods: 1,212 secondary metabolites from brown algae were screened using ADMET profiling. Compounds with favorable properties were selected for molecular docking against A179L, an antiapoptotic protein of ASFV. The top-scoring metabolites were further analyzed through molecular dynamics simulations, principal component analysis (PCA), and binding energy calculations.

Results: ADMET profiling identified compounds with drug-like pharmacokinetic properties. Consensus molecular docking using two docking programs predicted the binding poses of the metabolites, and the top ten candidate ligands with the strongest docking scores were selected. We then performed 300 ns molecular dynamics simulations to assess the stability of the protein-ligand complexes, which were further validated through Principal Component Analysis (PCA). Finally, binding energies were estimated using both MMPBSA and MMGBSA approaches, and the key residues involved during binding were identified. All ten final candidate ligands demonstrated stability and favorable binding affinity (-27.78 kcal/mol to -38.58 kcal/mol).

Conclusion: Five compounds (CID: 24796376, sargachromanol G, strictaepoxide, CID: 163107957, and saringosterone) with the strongest binding energies are recommended for in vitro and in vivo testing to validate their potential as therapeutic agents against ASFV.]]> https://www.benthamscience.com/article/1546532026-06-04 The increasing resistance to anti-TB drugs has become a global issue, highlighting an urgent need to investigate new drug candidates for combating Mycobacterium tuberculosis (M.tb.). Asiatic Acid (AA), a major phytoconstituent of Centella asiatica, exhibits anti-microbial and anti-biofilm activities against several microbes. In the present investigation, AA was explored for prospective anti-biofilm activity against mycobacteria.

Methods: We first used a computational approach to probe the in silico inhibitory potential of AA against selected target proteins of the FAS-II pathway, involved in mycolic acid biosynthesis, which contributes to mycobacterial cell wall and biofilm formation. Further, in vitro anti-mycobacterial assays were performed to determine the inhibitory concentration and biofilm inhibition against M. smegmatis, which was quantified by crystal violet staining and validated by SEM and CLSM.

Results: The structure-based inhibitory potential of AA was evaluated against β-Ketoacyl ACP Synthase (KasA) and Enoyl acyl carrier protein (InhA) by molecular docking analysis, showing good binding affinities, with binding energies of -9.53 kcal/mol and -10.53 kcal/mol, and inhibition constants of 3.04 μM and 80.20 nM, respectively. Further, the in vitro anti-mycobacterial assays confirmed the MIC as 20 μg/ml against M. smegmatis. A conspicuous reduction in mycobacterial biofilm formation was observed upon exposure to AA at the MIC, as quantified by crystal violet staining and validated by SEM and CLSM. The EC50 value, the concentration showing 50% inhibition of biofilm formation, was observed as 40 μg/ml against M. smegmatis.

Discusssion: This study shows advancement in understanding the natural compound AA as a potent anti-tubercular compound by elucidating its dual mechanism of action, including antimycobacterial and anti- biofilm activity, and by targeting enzymes of the FASII pathway, i.e., InhA and KasA. The present in vitro investigation suggests that asiatic acid may serve as a good anti- TB compound, with the possibility of potentiation of anti- mycobacterial effects via additional biofilm-inhibition activity.

Conclusion: These findings are expected to pave the way for the design and development of novel anti-TB medications and strategies that enhance treatment efficacy and minimize resistance development in this persistent pathogen. Further investigations are required to determine the mode of action and validate its candidacy as a promising anti-biofilm agent in the current clinical setting.]]> https://www.benthamscience.com/article/1555272026-06-04 Introduction: Alpha-Lipoic Acid (ALA) is known for its antioxidant and antiinflammatory properties; however, its direct role in neurogenesis remains undefined. This study evaluated the neurogenic potential of the R-enantiomer (R-ALA) using transcriptomic profiling and in vitro validation.

Methods: Transcriptome analysis of R-ALA-treated cells was performed to identify differentially expressed genes and enriched pathways. Validation was conducted in different cell lines (C6, PC12, TGW) to assess neurite morphology and neuronal marker expression.

Results: R-ALA induced strong upregulation of several neurogenesis-associated biomarkers and enriched pathways related to neurogenesis, axonogenesis, dendritogenesis, gliogenesis, and neurotransmission (serotonergic, GABAergic). In line with transcriptomic predictions, treated cells exhibited neurite outgrowth and increased expression of neuronal markers, including MAP2, β- tubulin III, NF-200, and GAP43.

Discussion: The upregulation of neuron-specific markers and downregulation of progenitorassociated CD44 suggest a comprehensive activation of neuroplastic and regenerative processes, reinforcing ALA’s potential as a neurogenic agent.

Conclusion: These findings provide the first integrated transcriptome-to-phenotype evidence that R-ALA promotes neurogenesis and neuronal differentiation in vitro, offering a basis for further in vivo exploration in neurodegenerative disorders.]]> https://www.benthamscience.com/article/1481402026-06-04 Background: Skin cancer is one of the most prevalent cancers globally and is considered a serious public health problem associated with high death rates. The current therapeutic regimes for skin cancer are limited by their low bioavailability, generation of resistance, or adverse side effects. Many fruit extract-based nutraceuticals hold potential as topical treatment methods. Pyrus communis (Pear) fruit extract is a rich source of cholinergic acid, presently used as therapy for various skin diseases. Thus, it qualifies as a promising candidate for skin cancer treatment.

Objective: The objective of the study is to evaluate the cytotoxicity of Pyrus communis extract entrapped in ethosomes.

Methods: In this study, Pyrus communis fruit extract was formulated in ethosomes using the hot method and optimized using central composite design. The optimized ethosomes were characterized in vitro for particle size distribution, zeta potential, entrapment efficiency, morphology, and particle stability.

Results: Preliminary phytochemical screening results suggest that PCHE contains a significant amount of phenolic compounds compared to other extracts (PCEA and PCAE). The presence of these phenolic compounds contributes to the strong antioxidant and cytotoxic effects of PCHE, which are observed in a dose-dependent manner. Analysis through GC-MS has identified chlorogenic acid, arbutin, ursolic acid, quercetin, and epicatechin are present in PCHE. Based on the initial testing of the extracts, PCHE was chosen for the preparation of ethosomes. The optimized ethosomes were found to have a particle size of 699 nm and a zeta potential of -16.07. Transmission Electron Microscopy illustrated a closed, spherically symmetrical structure of the ethosomes. Additionally, the Franz diffusion cell analysis for percutaneous absorption using egg membrane indicated a steady-state flux of the drug from the ethosomes. The formulation's cytotoxicity potential was assessed using the epidermoid carcinoma cell line (A431) through the MTT assay. The results show that the ethosome formulations exhibit cytotoxic activity better than PCHE extract. 1

Conclusion: In sum, the result of this study clearly points out that Pyrus communis extract entrapped in ethosomes, prepared by hot method, displayed a cytotoxic potential against skin cancer cell lines. This ethosomal formulation can be harnessed for skin cancer therapy through further mechanistic analysis and animal studies.]]> https://www.benthamscience.com/article/1526942026-06-04 Introduction: Ovarian cancer (OC) is a heterogeneous cancer with a high death rate and poor prognosis. Identifying precise and reliable prognostic biomarkers is crucial for the treatment of OC.

Methods: Anoikis-related DEGs were screened from the differentially expressed genes (DEGs) between the OC group and the control group and then subjected to functional enrichment analysis. A protein-protein interaction (PPI) network was developed to obtain candidate genes. Then, LASSO regression and support vector machine-recursive feature elimination (SVM-RFE) analysis were employed to select biomarkers, followed by conducting gene set enrichment analysis (GSEA). Correlations between the biomarkers and immune infiltration, drug sensitivity, and immunotherapy response were assessed by the Spearman method. The expression of the biomarkers in cells was measured by scRNA-seq analysis.

Results: We obtained 142 anoikis-related DEGs, which were mainly enriched in apoptosis-relevant pathways. A total of 16 candidate genes were acquired from the PPI network. Then, STAT3 and BCL2L1 were selected via LASSO regression and SVM-RFE analysis as two biomarkers for OC. BCL2L1 was closely associated with the infiltration of 5 immune cell types and 32 drugs, while STAT3 exhibited notable correlation with the infiltration of 6 immune cell types and 13 drugs. The Tumor Immune Dysfunction and Exclusion (TIDE) score was positively correlated with the two biomarkers. Moreover, STAT3 and BCL2L1 were expressed in most cells, with a high expression of STAT3 in endothelial cells.

Discussion: This study integrated bulk and single-cell transcriptomics to identify STAT3 and BCL2L1 as two anoikis-related biomarkers linked to the immune infiltration and drug sensitivity of OC, showing potential value for patient stratification and therapy. These findings suggested that targeting the STAT3/BCL-xL axis and combinational immunotherapy might be an effective strategy for OC treatment, which, however, should be further verified by functional and clinical experiments.

Conclusion: This study identified two anoikis-related biomarkers for OC, contributing to the clinical diagnosis of OC and its treatment.