Natural products are rich therapeutic sources, yet their translation into effective medicines remains challenging. Bee venom (BV) contains a diverse repertoire of bioactive molecules, but gentle, scalable methods to enhance its functionality are limited. This study, therefore, investigates a novel, solvent-free, post-extraction infrared (IR) conditioning step to fine-tune BV's composition and bioactivity. BV was irradiated (230 V, 50 Hz, 150 W) and compared to native extract using GC–MS and bioactivity assays. GC–MS revealed selective compositional tuning, with significant enrichment ( p ≤ 0.05) of 4H-1-benzopyran-4-one, 2-(3,4-dimethoxyphenyl)-3,5-dihydroxy-7-methoxy. IR-treated BV exhibited enhanced antimicrobial activity, with increased zones of inhibition for Staphylococcus aureus (15 ± 0.1 vs. 11 ± 0.4 mm), Bacillus subtilis (24 ± 0.2 vs. 22 ± 0.6 mm), Candida albicans (26 ± 0.1 vs. 22 ± 0.5 mm), Klebsiella pneumoniae (24 ± 0.4 vs. 15 ± 0.3 mm), and Salmonella typhi (27 ± 0.8 vs. 20 ± 0.7 mm). MICs decreased for S. aureus , B. subtilis , and K. pneumoniae . The treated BV also showed stronger antibiofilm activity at 25% MBC concentrations for K. pneumoniae and S. typhi ( p ≤ 0.05) and significantly reduced hemolysis at 25% MIC for S. aureus and B. subtilis ( p ≤ 0.05). Antioxidant capacity increased (DPPH IC ₅₀ : 16.47 ± 1.1 vs. 27.65 ± 0.8 µg/mL), as did anti-inflammatory activity (COX-2 IC ₅₀ : 48.84 ± 0.2 vs. 50.99 ± 0.9 µg/mL; COX-1 IC ₅₀ : 24.7 ± 0.2 vs. 41.74 ± 0.2 µg/mL). Cytotoxicity against PC-3 and SKOV-3 cells was maintained (IC ₅₀ : 19.73 ± 0.9 and 19.76 ± 0.11 µg/mL, respectively, vs. native BV's 11.48 ± 0.3 and 17.46 ± 0.27 µg/mL). These findings establish brief IR irradiation as a practical, scalable post-processing strategy to selectively enhance the therapeutic potential of BV for biomedical applications. Graphical abstract

Ozonation is an emergent green technology that modifies the chemical composition and bioactivity of natural oils, creating new opportunities for functional and biomedical use. In this study, the chemical changes and in vitro activities of lettuce (Lactuca sativa) oil before and after ozonation were evaluated. Gas chromatography–mass spectrometry (GC–MS) revealed an increase in both the number and diversity of constituents in ozonated oil, with (Z)-13-docosenamide and trans-13-octadecenoic acid as predominant components. Fourier-transform infrared (FTIR) spectra showed overall similarity between native and ozonated oils, but with three additional characteristic bands in the ozonated sample. Bioassays demonstrated that ozonation enhanced anti-Helicobacter pylori activity (inhibition zone 21.3 ± 0.3 mm), supported bactericidal effects, and improved antibiofilm and antihemolytic properties. The antioxidant capacity of ozonated oil was modestly increased (IC₅₀ = 3.95 ± 0.4 µg/mL), while butyrylcholinesterase inhibition was more markedly enhanced (IC₅₀ = 2.58 ± 0.6 µg/mL), compared to that of the non-ozonated oil (IC₅₀ = 6.14 ± 0.3 µg/mL and IC₅₀ = 4.38 ± 0.4 µg/mL, respectively). Molecular docking suggested strong interactions of major ozonation-derived compounds with human BuChE and H. pylori urease, providing mechanistic support for the observed activities. Overall, these results indicate that ozonation modestly but consistently enhances the biological potential of lettuce oil through compositional shifts, highlighting its promise for development as a safe functional food ingredient with possible biomedical applications.

Background/Objectives: Arsenic (ARS) exposure is a major cause of kidney injury, driven by oxidative stress, inflammation, fibrosis, and apoptosis. This study evaluated the renoprotective effects of morin (MOR) and morin-loaded PLGA nanoparticles (MOR–PGNPs) against ARS-induced nephrotoxicity in rats. Methods: Sixty male Sprague Dawley rats were randomly allocated into six groups (n = 10 per group). The control group received corn oil. The MOR group received MOR (100 mg/kg), and the MOR–PGNPs group received the same dose of MOR encapsulated in PLGA nanoparticles. ARS was administered at 10 mg/kg for 14 days. Co-treated groups received ARS together with either MOR or MOR–PGNPs, with a 28 min interval between administrations. Renal function markers (serum urea, creatinine, uric acid, renal KIM-1), oxidative stress and antioxidant parameters (Nrf2/HO-1, CAT, SOD, GPx, ROS, MDA), inflammatory mediators (TLR4/NF-κB, TNF-α, IL-6, IL-1β), fibrotic markers (TGF-β1, fibronectin), and apoptotic proteins (caspase-3, caspase-8, Bax, Bcl-2) were assessed, alongside histopathological and ultrastructural evaluations. Results: ARS exposure significantly impaired renal function, increased KIM-1, suppressed Nrf2/HO-1 signaling, reduced antioxidant enzyme activities, and elevated ROS and MDA levels. It also activated TLR4/NF-κB signaling, upregulated pro-inflammatory cytokines and fibrotic markers, and increased pro-apoptotic proteins while downregulating Bcl-2. MOR co-treatment partially ameliorated these alterations. MOR–PGNPs produced potentially enhanced protection, restoring kidney function markers, enhancing antioxidant defenses, and markedly attenuating inflammation, fibrosis, and apoptosis. Histopathological and ultrastructural analyses confirmed preservation of glomerular and tubular architecture, mitochondrial integrity, and minimal cytoplasmic vacuolization in the MOR–PGNPs group. Conclusions: MOR–PGNPs at 100 mg/kg effectively mitigated ARS-induced renal damage through antioxidant, anti-inflammatory, antifibrotic, and anti-apoptotic mechanisms, supporting PLGA-based morin nanoparticles as a promising and safe renoprotective strategy.

Background/Objectives: Medicinal plants are an abundant source of bioactive molecules, particularly in arid environments, such as Saudi Arabia, where Ocimum basilicum L. (Saudi basil) has long been used for its therapeutic properties. This study aimed to examine the phytochemical profile and bioactivities of non-ozonated (untreated) and ozonated methanolic extracts of O. basilicum and to determine whether ozonation enhances their biological effects, with a focus on antidiabetic, anti-Alzheimer, anti-inflammatory, antimicrobial, and cytotoxic properties. Methods: Fresh leaves of O. basilicum were extracted with methanol, subjected to ozonation, and analyzed by HPLC. In vitro assays were conducted to evaluate α-amylase, α-glucosidase, and BChE inhibition, RBC membrane stabilization, antibacterial activity against Helicobacter pylori and cytotoxicity using normal lung fibroblasts (WI-38) and human colorectal adenocarcinoma cell line (Caco-2). Results: Ozonation modified the phytochemical profile, enriching chlorogenic and rosmarinic acids. Ozonated extracts exhibited stronger inhibition of α-amylase with an IC₅₀ of 5.09 µg/mL compared to 13.6 µg/mL of untreated Saudi basil and α-glucosidase (IC₅₀ 6.15 µg/mL vs. 9.42 µg/mL). They also showed enhanced BChE inhibition with an IC₅₀ of 13.4 µg/mL compared to 31.8 µg/mL of non-ozonated extract. In addition, ozonated extracts produced significant anti-inflammatory effects by stabilizing RBCs, with an IC₅₀ of 8.04 µg/mL compared to 8.44 µg/mL for untreated extracts and 4.41 µg/mL for indomethacin. Ozonated extracts produced larger H. pylori inhibition zones (26.7 mm) and an MBC/MIC ratio of 1. Cytotoxicity testing revealed that ozonated extracts were less toxic to WI-38 cells, with IC₅₀ values of 437.89 µg/mL versus 191.06 µg/mL, and 149.14 µg/mL compared to 103.7 µg/mL of untreated Saudi basil in Caco-2 cells. Conclusions: Ozonation enriches the phytochemical composition of O. basilicum, enhancing antidiabetic, neuroprotective, anti-inflammatory, and antibacterial activities while reducing cytotoxicity on normal cells. These findings support the potential of ozonated O. basilicum as a safe and promising natural therapeutic candidate for metabolic, neurodegenerative, and infectious diseases.

This study explored the impact of ethanol as a co-solvent in supercritical fluid extraction on the recovery of bioactive compounds from carrot seeds and assessed the resulting extracts for antimicrobial, α-amylase and α-glucosidase, and butyrylcholinesterase inhibitory potential. Ethanol supplementation significantly improved extraction performance, with the yield increasing from 110 mg in the absence of ethanol to 134 mg at 5% ethanol, followed by a slight decrease to 132 mg at 10%. High-performance liquid chromatography (HPLC) revealed pronounced phytochemical enrichment at 5% ethanol, particularly for chlorogenic acid (1541.24 µg/g), gallic acid (1279.27 µg/g), and hesperetin (1513.68 µg/g), indicating enhanced recovery of phenolic and flavonoid constituents. The 5% ethanol extract demonstrated superior antimicrobial activity, producing inhibition zones of 19 mm against Enterococcus faecalis, 26 mm against Klebsiella pneumoniae, 25 mm against Staphylococcus aureus, and 29 mm against Candida albicans. Values of both minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were markedly reduced, while antibiofilm activity reached 93.11% for E. faecalis and 91.00% for K. pneumoniae. The extract also exhibited potent inhibitory effects with IC₅₀ values of 7.74 and 13.37 µg/mL, against α-amylase and α-glucosidase, correspondingly, as well as strong butyrylcholinesterase inhibition (IC₅₀ = 2.51 µg/mL), highlighting promising α-amylase/α-glucosidase and butyrylcholinesterase inhibitory potential. Molecular docking further supported these findings, showing that chlorogenic acid bound more strongly than vanillin to OmpK36, lysosomal acid-α-glucosidase, and butyrylcholinesterase, with docking scores ranging from −6.1 to −6.9 kcal/mol. These findings identify ethanol-modified supercritical fluid extraction as a sustainable and effective green strategy for improving the recovery of carrot seed bioactives and enhancing their multifunctional in vitro biological properties. Notably, this study provides the first comprehensive evidence that 5% ethanol modification selectively enriches key phenolic constituents, including chlorogenic acid, gallic acid, and hesperetin, in carrot seed extracts, with corresponding enhancement of α-amylase, α-glucosidase, and butyrylcholinesterase inhibitory activities.

Natural products may address infection, oxidative stress and metabolic disorders. This study used gas chromatography-mass spectrometry (GC-MS), bioassays and molecular docking to characterize Delonix regia seed oil and evaluate its antimicrobial, antibiofilm, radical-scavenging and pancreatic lipase-inhibitory activities. The most abundant tentatively identified peaks in the GC-MS total ion chromatogram were β-sitosterol (26.35%), stigmasterol (14.57%), linoleic acid (10.39%), and oleic acid (5.82%). Minimum inhibitory concentrations (MICs) were 15.62 µg/mL for Listeria monocytogenes, Salmonella typhi, and Candida albicans, and 62.5 µg/mL for methicillin-resistant Staphylococcus aureus. The corresponding inhibition zones were 28 ± 0.7, 21 ± 0.3, 31 ± 0.1, and 15 ± 0.5 mm, respectively. At 75% of the minimum bactericidal/fungicidal concentration, the oil inhibited biofilm formation by 94.95% for both L. monocytogenes and C. albicans. DPPH and ABTS radical-scavenging IC₅₀ values were 17.04 ± 0.25 and 13.17 ± 0.40 µg/mL, respectively, while the pancreatic lipase IC₅₀ was 26.29 ± 0.08 µg/mL. Time-kill assays demonstrated ≥3 log₁₀ CFU/mL reductions, with no recoverable colonies for any of the five tested pathogens within 3 h. Docking predicted stronger stigmasterol binding to L. monocytogenes Sortase A (−6.60 kcal/mol) than to C. albicans lanosterol 14α-demethylase (CYP51; −3.41 kcal/mol), involving hydrogen bonds with Met63 and Arg381, respectively, and hydrophobic contacts. These findings identify a phytosterol-enriched GC-MS-detectable fraction of Delonix regia seed oil with in vitro activities warranting further mechanistic and in vivo investigation.