Caffeine induces concentration-dependent biphasic or hormetic effects in insects, yet its impact on Drosophila ananassae remains unexplored. We evaluated adult physiological traits (survival, fecundity, lipid content) under adult-only exposure and developmental traits under combined short-term parental adult and continuous offspring developmental exposure across a caffeine concentration gradient (0.6-4.8 mg/ml). Low concentrations (0.6-1.8 mg/ml) increased survival (LT50) approximately 1.6-fold over control (Kaplan-Meier; log-rank P < 0.01), moderately increased fecundity and adult lipid content. Proportions of pupae eclosion were near control, and eggs hatched were slightly but significantly higher than control (P < 0.05) at lower concentrations. Higher concentrations (≥ 3.0 mg/ml) were detrimental, decreasing survival (̴2.5-folds), fecundity, and lipid content, and progressively impairing performance. The proportion of pupae eclosion declined significantly beyond ≥3.6 mg/ml. Developmental duration increased for 2nd and 3rd larvae and pupae as concentration increased, indicating inhibition of post-embryonic development, whereas early embryonic and first-instar durations were unaffected. Mixed-model ANCOVA indicated significant treatment, isofemale line, and treatment × line interaction (P < 0.001), whereas dry body weight remained a non-significant covariate. After body size standardization, lipid content remained positively associated with fecundity across isofemale lines (R2 = 0.88-0.91, P < 0.001). Hormesis Index demonstrated stimulation for adult traits at low concentrations and inhibition at higher concentrations, with transition near 1.8 mg/ml. API showed improved integrated adult performance at lower concentrations, whereas DPI remained near control-equivalent levels or was slightly higher before both indices declined beyond 3.6 mg/ml. These findings suggest concentration-dependent caffeine hormesis, with lower concentrations increasing adult traits and maintaining near biological-maximum developmental performance under combined exposure, whereas higher concentrations impair physiological and developmental traits.

Although the adsorption of pharmaceuticals by microplastics (MPLs) may reduce environmental contaminant concentrations, their effects on bioavailability and biological toxicity following ingestion remain poorly studied. To mimic environmentally relevant exposure, environmentally relevant polypropylene microplastics (PP-MPLs) derived from post-consumer plastic products (real-life) were used as carriers for the pharmaceuticals paracetamol (PCM) and ciprofloxacin (CFX), using Drosophila larvae in vivo model. Free PCM and CFX were tested at concentrations equivalent to their respective adsorption loadings on PP-MPLs (0.03% and 0.02% w/w, respectively), enabling direct comparison under environmentally realistic conditions. PP-MPLs, with or without adsorbed pharmaceuticals, retained their irregular morphology and heterogeneous size distribution, representing an environmentally relevant mixture of micro- and nanoscale particles. Following oral exposure, PP-MPLs, either alone or carrying adsorbed pharmaceuticals, sequentially traversed the intestinal barrier penetrating the peritrophic membrane, accumulating within enterocytes, and ultimately reaching hemolymph. Particle exposure induced extensive enterocyte vacuolation, significantly increased intracellular reactive oxygen species (ROS), and triggered targeted gene-expression changes involving antioxidant, immune, detoxification, and stress-response pathways, while genes associated with digestive function were consistently deregulated. Despite elevated oxidative stress, PP-MPLs did not alter mitochondrial membrane potential. In contrast, free PCM induced significant mitochondrial depolarization without increasing ROS, whereas this effect was abolished following adsorption onto PP-MPLs, potentially due to reduced bioavailability. Moreover, pharmaceutical-loaded PP-MPLs did not elicit additive molecular toxicity compared with PP-MPLs alone.

Acetaminophen (APAP)-induced liver injury (AILI) is a common serious adverse drug event, yet its molecular mechanisms remain incompletely understood. To identify key regulators in AILI, we conduct a temporal liver transcriptomic sequencing analysis in mice subjected to APAP exposure, revealing a significant and dynamic alteration in the expression of immediate early response 2 ( Ier2). Single-cell RNA sequencing analysis further delineates Ier2 expression patterns across hepatic cell populations following APAP insult. Hepatocyte-specific Ier2 knockout (HKO) mice and hepatocyte-directed Ier2 overexpression mice are generated to elucidate the molecular function of IER2 in AILI. RNA sequencing and proteomics are performed to investigate the underlying protective mechanisms of IER2 in AILI. We demonstrate that hepatocyte-specific Ier2 deletion increases susceptibility to AILI, whereas hepatic Ier2 overexpression ameliorates APAP-induced liver injury. Integrated transcriptomic and proteomic analyses identify lipid metabolism as a key pathway modulated by IER2 overexpression, with ACOT2 as a putative target. Functional rescue experiments reveal that ACOT2 knockdown markedly attenuates the protective effect of IER2, indicating that ACOT2 is required for IER2-mediated hepatoprotection. In conclusion, IER2 protects against AILI by modulating lipid metabolism pathways, with ACOT2 as a critical downstream effector. These findings position IER2 as a promising and novel therapeutic target for mitigating AILI.

This study presents a novel approach combining stimulated Raman scattering (SRS) microscopy with mechanistic modelling to quantify the disposition and permeation of topically applied compounds in skin. The dermal delivery of 4-cyanophenol and caffeine was investigated using SRS microscopy, generating high-resolution images resolving their distribution within the stratum corneum (SC) lipid matrix and corneocytes. The imaging data revealed distinct permeation pathways, with 4-cyanophenol preferentially localising within the intercellular lipid matrix and caffeine within the corneocyte-rich regions. Depth-resolved SRS imaging provided concentration profiles across the SC, showing a monotonic decrease in 4-cyanophenol concentration, whereas caffeine exhibited depth-dependent transport behaviour. These experimental concentration profiles were modelled using a two-dimensional mechanistic model with calibration to reveal the diffusion, partition and deposition behaviour within the lipid and corneocyte domains. For 4-cyanophenol, the optimised permeability was consistent with literature-reported values and baseline model predictions, while for caffeine, optimisation substantially improved permeability predictions. Simulations further captured formulation-dependent effects predicting 1.3-fold and 1.5-fold increases in SC uptake of 4-cyanophenol and caffeine, respectively, in propylene glycol relative to aqueous vehicles. Overall, this work demonstrates the potential of combining SRS microscopy with inverse mechanistic modelling to derive transport parameters directly from skin imaging data and reveal dermal drug disposition and permeation. To our knowledge, this represents the first application of SRS-derived depth profiles to inform mechanistic modelling of skin permeation.

Improving the aqueous solubility of moderately soluble drugs using safe and sustainable solvent systems remains an important challenge in pharmaceutical formulation. In this study, a series of aqueous natural deep eutectic solvents (NADESs) were systematically designed and evaluated to enhance the solubility, stability, and biocompatibility of acetaminophen (ACP). The effects of hydrogen bond donor type, hydrogen bond acceptor type, molar ratio, and NADES concentration were investigated to identify the optimal formulation. Among the tested systems, the aqueous betaine chloride-acetic acid NADES (BeCl-Ace) at a molar ratio of 1 : 4 and a concentration of 70% exhibited the highest solubilization capacity, increasing ACP solubility by 25.28 and 22.14-fold at 25 and 37 °C, respectively, compared with water. Computational analyses based on electrostatic potential mapping, binding energy calculations, and interaction region indicator analysis revealed that the enhanced solubility originates from cooperative hydrogen bonding, electrostatic attraction, and van der Waals interactions between ACP and the BeCl-Ace components, particularly BeCl. In addition, the selected NADES formulation significantly improved ACP stability under thermal stress, retaining 89.13% of the initial drug content after 30 days at 37 °C, compared with 70.20% in water. Cytotoxicity evaluation on HEK-293A cells further confirmed good biocompatibility, with cell viability remaining above 92% across the tested concentration range. These findings indicate that aqueous BeCl-Ace NADESs represent a promising and sustainable solvent system for improving ACP formulation performance.

Pu-erh tea occupies an important position in tea consumption because of its distinctive flavor and potential health benefits; however, its flavor acceptability still has room for improvement. To enhance sensory quality and diversify product forms, this study investigated a blending system of American ginseng powder and ripe Pu-erh tea powder and compared the quality and chemical composition of samples with different blending ratios by integrating sensory evaluation, physicochemical analysis, flavoromics, and untargeted metabolomics. The results showed that SP2, consisting of 75% ripe Pu-erh tea powder and 25% American ginseng powder, achieved the highest overall sensory score among the experimental blends (SP1-SP3) and was characterized by an aged aroma accompanied by a chrysanthemum-like note, a mellow and full-bodied taste, and a pronounced sweet aftertaste. As the proportion of American ginseng increased, the contents of caffeine and esterified catechins decreased, whereas the soluble sugar content increased. Tentatively identified 2-methylbutanal and (E)-2-octenal in SP2 were considered potential aroma-active candidates. A total of 1,171 tentatively annotated non-volatile metabolic features were obtained, with representative differential features involving flavonoids, phenylpropanoid-related compounds, and ginsenoside-related components. These compositional differences mainly reflected the respective contributions of the two raw materials and changes in their blending proportions, providing a reference for formulation optimization and quality evaluation of blended products.