Plant-derived polysaccharides are critical bioactive compounds; however, conventional extraction methods are often inefficient, energy-intensive, and may compromise their bioactivity. Ultrasound-assisted deep eutectic solvent (UA-DES) extraction offers a greener alternative by integrating acoustic cavitation with tunable solvent properties; however, optimization remains complex due to the interaction of multiple processing variables. This study reports a novel application of ultrasound-assisted deep eutectic solvent (UA-DES) extraction for okra polysaccharides (OPs), with process optimization using response surface methodology (RSM) and artificial neural network (ANN) modeling to identify optimal conditions and clarify nonlinear extraction behavior. Among the tested DES systems, choline chloride-citric acid (CCA) exhibited the highest extraction performance. Single-factor experiments and RSM identified sonication time and liquid-solid ratio as key variables. The ANN model achieved higher predictive accuracy than RSM and captured nonlinear and synergistic parameter interactions that were not evident in traditional response surfaces, providing deeper insight into process behavior. Under optimized conditions (2 h, 80 °C, 190 W, 60 mL/g), UA-DES extraction produced 23.56 % OPs and 80.75 % DPPH• scavenging activity, representing 94 % higher yield and 28 % greater antioxidant activity than hot-water ultrasonic (HWU) extraction. UA-DES-derived OPs contained higher contents of uronic acids, total sugars, and glucans, and uniquely included arabinose absent in HWU extracts. Structural analyses revealed pyranose configurations, amorphous crystallinity, and porous microstructures, which collectively contribute to improved solubility and bioactivity. Overall, UA-DES extraction using CCA provides an eco-efficient strategy for producing high-value okra polysaccharides. The integrated RSM-ANN framework enables precise optimization and enhanced mechanistic understanding, supporting UA-DES as a scalable, green technology for the production of functional polysaccharides.

Alzheimer’s disease (AD), a leading neurodegenerative disorder, is closely associated with the accumulation of amyloid-beta (Aβ) peptides in the brain. The enzyme β-secretase (BACE1), pivotal in Aβ production, represents a promising therapeutic target for AD. While bioactive peptides derived from food protein hydrolysates have neuroprotective properties, their inhibitory effects on BACE1 remain largely unexplored. In this study, we evaluated the inhibitory potential of protein hydrolysates from gliadin, whey, and casein proteins prepared using bromelain, papain, and thermolysin. Through in vitro and cellular assays, bromelain-hydrolyzed gliadin (G-Bro) emerged as the most potent BACE1 inhibitor, with an IC₅₀ of 0.408 mg/mL. G-Bro significantly reduced BACE1 expression and amyloid precursor protein (APP) processing in N2a/PS/APP cell cultures, suggesting its potential to attenuate Aβ aggregation. The unique peptide profile of G-Bro likely contributes to its inhibitory effect, with proline residues disrupting β-sheets, lysine residues introducing positive charges that hinder aggregation, hydrophobic residues stabilizing binding interactions, and glutamine residues enhancing solubility and stability. These findings highlight gliadin hydrolysates, particularly G-Bro, as potential natural BACE1 inhibitors with applications in dietary interventions for AD prevention. However, further studies are warranted to elucidate specific peptide interactions and their bioactivity in neural pathways to better understand their therapeutic potential.

Polycyclic aromatic hydrocarbons (PAHs), carcinogenic substances primarily formed through pyrolysis and oxidation of fat at high cooking temperatures, are commonly found at high levels in grilled meats. Reducing PAHs formation by incorporating natural antioxidants, such as through marination, has been demonstrated to be effective. However, the inhibitory effect of fresh phenolic-rich fruit powders on PAHs formation in charcoal-grilled meats remains unknown. To clarify the application of the fruit powders, 15 experimental groups were conducted. All pretreatment techniques (spraying, marinating, and mixing) were applied across all four freeze-dried fruit powders (lemon, guava, papaya, and mango). Each method was systematically tested with each fruit powder to evaluate its effect on inhibiting the formation of the four PAHs (BaA, CHR, BbF, and BaP) in charcoal-grilled pork belly and loin. Firstly, guava powder exhibited the highest phenolic content and antioxidant activity compared to the lemon, papaya, and mango powders (p < 0.05), among which the main phenolic compounds were ellagic acid, quercetin, and epigallocatechin gallate (EGCG). Further, marination of pork belly with guava powder exhibited the highest inhibition rate of PAHs (94.8%), followed by lemon (91.1%), papaya (89.8%), and mango (89.0%), with a statistically significant difference at p < 0.05. The reduction in estimated daily intake (EDI) and the increase in the margin of exposure (MOE) indicate that consuming grilled meat treated with these fruit powders poses no safety concerns and may potentially reduce health risks. Finally, the sensory evaluation showed that marinating with guava powder did not perceptibly affect the sensory attributes of the meat. Overall, this study provides a potent strategy for inhibiting the formation of PAHs in meat during charcoal grilling by incorporating fruit powder while preserving sensory qualities.

Type 2 diabetes mellitus remains a critical global health challenge, driving the pursuit of novel therapeutic strategies. This study investigated the anti-diabetic efficacy of the peptide 1CBR, derived from sodium caseinate hydrolysate, administered orally at 25 mg/kg/day to db/db mice over a 4-week period. Glucose tolerance was evaluated via oral glucose tolerance tests (OGTT), while plasma dipeptidyl peptidase-IV (DPP-IV) activity, glucagon-like peptide-1 (GLP-1), and insulin concentrations were quantified using enzyme-linked immunosorbent assays (ELISA). Two bioactive peptides, GPFPLPD and APDSGNFR, were isolated and characterized, exhibiting half-maximal inhibitory concentrations (IC₅₀) of 99.12 µM and 73.07 µM for DPP-IV inhibition, respectively, and both significantly stimulated GLP-1 secretion in enteroendocrine cells in vitro. Pharmacokinetic analysis in Sprague–Dawley rats demonstrated oral bioavailability of 11.28% and 19.12% for these peptides, surpassing typical expectations for peptide-based agents. Collectively, these results provide compelling evidence that 1CBR-derived peptides exert glucose-lowering effects through the dual mechanisms of DPP-IV inhibition and GLP-1 stimulation, combined with favorable oral absorption profiles. These findings underscore the potential of 1CBR peptides as promising candidates for development into nutraceuticals or pharmaceutical agents for diabetes management.

Fluoxetine (FXT) and alprazolam (APZ), widely used for mental disorders, have poorly studied adverse effects on mitochondrial function, including oxidative phosphorylation, electron transport, and membrane permeability. This study represents the first investigation using a chick embryo model (HH-stage 10, day 1.5) to analyze the teratogenic effects of FXT and APZ and explore the protective potential of coenzyme Q10 (CoQ10) and L-carnitine (CNT). Administration of FXT (10 μM) and APZ (1 μM) resulted in high teratogenic rates of 53 % and 80 %, respectively, predominantly manifesting as lipid myopathy in hatching muscles, characterized by lipid accumulation, myofibril disruption, inflammation, and edema. Gene expression analysis revealed upregulation of acetyl-CoA carboxylase (ACC) and downregulation of carnitine palmitoyltransferase 1 (CPT1), leading to impaired lipid peroxidation and excessive reactive oxygen species (ROS) production. Markers of oxidative stress, including superoxide dismutase (SOD), hydrogen peroxide (H₂O₂), and nitric oxide (NO), were significantly elevated, correlating with glutathione (GSH) depletion and mitochondrial ultrastructural damage, resulting in reduced ATP production. Notably, co-administration of CoQ10 and CNT with FXT or APZ significantly improved teratogenic and mortality rates and reduced oxidative stress levels. Specifically, CoQ10 (2 μM) in the FXT group significantly reduced SOD, H₂O₂, and NO levels, while co-treatment with CNT and CoQ10 (2 μM) in the APZ group significantly alleviated NO levels. This pioneering study highlights the novel and crucial potential of CoQ10 and CNT as nutritional supplements to mitigate mitochondrial damage and antioxidant system imbalance caused by FXT and APZ, providing an innovative strategy for clinical application.