Medicinal plants have traditionally been used to treat various human diseases worldwide. In this study, we evaluated the leaf extracts of plants from the Acanthaceae family, specifically Clinacanthus nutans (Burm.f.) Lindau, Thunbergia laurifolia Lindl., and Acanthus ebracteatus Vahl., for their compounds and antioxidant activity. The ethanolic extracts of A. ebracteatus showed the highest total phenolic content at 22.55 mg GAE/g extract and the strongest antioxidant activities, with IC₅₀ values of 0.24 mg/mL and 3.05 mg/mL, as determined by DPPH and ABTS assays. The antibacterial efficacy of these extracts was also tested against Streptococcus pyogenes, Streptococcus mutans, Staphylococcus aureus, and Klebsiella pneumoniae. The diameters of the inhibition zones ranged from 14.7 to 17.3 mm using the agar well diffusion method, with MIC and MBC values ranging from 7.81 to 250 mg/mL. Anti-biofilm formation, antibacterial adhesion, and antibacterial invasion assays further demonstrated that these medicinal plant extracts can inhibit bacterial biofilm formation and prevent the adhesion and invasion of oral pathogenic bacteria on the human tongue squamous cell carcinoma-derived cell line (HSC-4 cells). The ethanolic extracts of C. nutans and A. ebracteatus were able to inhibit the gtfD and gbp genes, which facilitate biofilm formation and bacterial adherence to surfaces. These findings provide new insights into the antibacterial and antioxidant properties of plant extracts from the Acanthaceae family. These activities could enhance the clinical and pharmaceutical applications of plant extracts as an alternative therapy for bacterial infections and a dietary supplement.

Arthrospira platensis (spirulina) is a cyanobacterium containing various phytochemical compounds associated with various antioxidant, antimicrobial, antiviral, anticancer, anti-inflammatory, and immune-promoting properties. The efficacy of ethanolic and methanolic crude extracts of A. platensis regarding antibacterial, antioxidant, and anticancer effects was determined in this study. The ethanolic extract showed the highest antioxidant activity by 8.96 ± 0.84 mg gallic acid equivalent per gram of extract (mg GAE/g extract), 53.03 ± 4.21 mg trolox equivalent antioxidant capacity per gram of extract (mg TEAC/g extract), and 48.06 ± 0.78 mg TEAC/g extract as determined by DPPH, ABTS, FRAP assays, respectively. Moreover, the ethanolic extract showed the highest total phenolic and flavonoid compound contents by 38.79 ± 1.61 mg GAE/g extract and 27.50 ± 0.53 mg of quercetin equivalent per gram of extract (mg QE/g extract). Gallic acid and quercetin in the extracts were also determined by HPLC. The antibacterial activity was evaluated by agar well diffusion and broth dilution methods on skin pathogenic bacteria, including Staphylococcus aureus, Staphylococcus epidermidis, methicillin-resistant S. aureus (MRSA), Micrococcus luteus, Pseudomonas aeruginosa, and Cutibacterium acnes. The inhibition zone of A. platensis extracts ranges from 9.67 ± 0.58 to 12.50 ± 0.50 mm. In addition, MIC and MBC values ranged from 31.25 to 125 mg/mL. The inhibition of biofilm formation and biofilm eradication by A. platensis ethanolic extract was 87.18% and 99.77%, as determined by the crystal violet staining assay. Furthermore, the anticancer activity of A. platensis on A375 human melanoma cells was examined. The ethanolic and methanolic extracts induced DNA apoptosis through both intrinsic and extrinsic pathways by upregulating the expression of caspase-3, caspase-8, and caspase-9. These findings suggested that A. platensis demonstrated promising antioxidant, antibacterial, and anticancer activities, emphasizing its potential as a natural therapeutic agent for the management of pathological conditions.

Background: Iron deficiency remains a major nutritional challenge, partly due to the limited stability and bioavailability of conventional iron formulations in foods and during digestion. In this study, iron–protein microparticles (IP-MPs) based on bovine serum albumin (IA-MPs) and hemp protein (IH-MPs) were developed via coprecipitation and evaluated as food-compatible iron delivery systems. Methods: Iron–protein microparticles (IP-MPs) were fabricated by a coprecipitation technique. The stability of IP-MPs was investigated in a three-phase digestion model. The uptake of IP-MPs by Caco-2 cells as well as the Ferritin concentration in Caco-2 cells were investigated. Results: Particle morphology and size distribution were strongly dependent on the protein matrix, with hemp protein microparticles exhibiting greater size uniformity and higher stability under simulated gastric conditions. In a standardized in vitro gastrointestinal digestion model, both IP-MP formulations preserved iron predominantly in the bioactive Fe(II) state and remained sufficiently intact to reach the intestinal phase. Biocompatibility and iron uptake were assessed using Caco-2 cell monolayers. Neither formulation induced cytotoxic effects, while iron delivered via IP-MPs showed enhanced cellular uptake compared to a commercial iron supplement and ferrous sulfate. The amount of Fe(II) detected in the basolateral compartment of IH-MP and IA-MP samples (1.4 µg and 1.3 µg, respectively) was higher than that observed for Floradix^® samples (approximately 0.7 µg) and corresponded to about 25% of the total iron applied. Functional iron bioavailability, assessed by ferritin formation, was significantly higher for IP-MPs, with hemp protein microparticles yielding the strongest ferritin response. Conclusions: These results demonstrate that iron–protein microparticles, particularly those based on hemp protein, effectively improve iron stability during digestion and enhance cellular iron bioavailability, highlighting their potential for application in iron fortification and functional food systems.

Bovine mastitis, an inflammation of the mammary gland, is recognized as the most prevalent disease in dairy cattle, leading to significant economic losses due to the reduction in quality and yield of milk. While antibiotic treatment remains to be the primary control method but their use of antibiotics contributes to the emergence of antibiotic-resistant bacteria and possesses potential health risks to consumers. Enhancement of host immune responses represents a promising alternative strategy for combating pathogenic bacteria. This study aimed to evaluate the immunomodulatory potential of metabolites derived from lactic acid bacteria (LAB) in murine macrophage RAW264.7 cells infected with Staphylococcus haemolyticus, a causative agent of bovine mastitis. Notably, LAB-derived metabolites demonstrated significant, strain-specific immunostimulatory activity. A comparative metabolomic analysis confirmed that each strain possessed a unique metabolic profile, providing a chemical basis for these various responses. The most pronounced effects were observed with metabolites from the isolated strain Enterococcus faecalis, which markedly increased NO production. Furthermore, these metabolites upregulated the expression of key inflammatory genes, e.g., iNOS, COX-2, TNF-α, and IL-6 and enhanced the protein levels of iNOS and COX-2. These findings suggest that LAB metabolites, particularly those from E. faecalis, may offer a novel therapeutic approach for enhancing immune defenses against mastitis-causing pathogens.

Thai medicinal flowers, namely Mesua ferrea L. (Bunnak), Mammea siamensis T. Anderson (Saraphi), and Clitoria ternatea (Anchan) have long been valued for their traditional medicinal. This study investigated their phytochemical composition and bioactivities, with a particular focus on antioxidant and antibacterial properties. Methods: Ethanolic flower extracts were analyzed by high-performance liquid chromatography (HPLC) and liquid chromatography–mass spectrometry (LC–MS). Antioxidant activities were determined by DPPH, ABTS, and FRAP assays. Antibacterial activity against Escherichia coli, E. coli O157:H7, Salmonella Typhi, Shigella dysenteriae, and Vibrio cholerae were assessed by agar well diffusion, broth dilution methods, and time–kill assays. Biofilm formation, biofilm disruption, and bacterial adhesion to Caco-2 cells were evaluated. Morphological changes in E. coli O157:H7 were examined using scanning electron microscopy (SEM), and leakage of intracellular contents (DNA, RNA, proteins) were quantified. Results: HPLC analysis revealed the highest level of gallic acid in M. ferrea and quercetin in M. siamensis. LC–MS analysis identified fifteen putative metabolites across the flower extracts, including quercetin, kaempferol, catechin, and luteolin derivatives, with species-specific profiles. C. ternatea extract exhibited the greatest total flavonoid content and antioxidant activity. Among the extracts, M. ferrea exhibited the strongest inhibitory effect, with inhibition zone of 13.00–15.00 mm and MIC/MBC values of 31.25–62.5 mg/mL. All extracts exhibited time-dependent bactericidal activity, significantly inhibited biofilm formation, disrupted established biofilms, and reduced bacterial adhesion to intestinal epithelial cells. SEM revealed membrane disruption in E. coli O157:H7 and leakage of intracellular components. Conclusions: Thai medicinal flower extracts, particularly M. ferrea, possess strong antioxidant and antibacterial activities. Their ability to inhibit biofilm formation, interfere with bacterial adhesion, and disrupt bacterial membranes highlights their potential as natural alternatives for preventing or controlling enteric bacterial infections.

Propolis is a resinous natural product collected by bees from plant sources to protect the hive. Its chemical composition varies according to bee species and environmental conditions, resulting in differences in its biological activities. This study aimed to evaluate the biological properties of stingless bee propolis collected from different provinces in Thailand, with particular attention to two native species, Tetragonula pegdeni and Tetragonula laeviceps. These two species of Tetragonula are commonly found in tropical regions of the country. The investigation focused on their antioxidant, antimicrobial, and anti-inflammatory activities. The propolis extract from T. laeviceps stingless bee (sample D) exhibited the highest total phenolic and flavonoid contents, at 29.59 mg GAE/g extract and 73.95 mg QUE/g extract, respectively. Among the propolis extracts, T. pegdeni stingless bee propolis extract (sample A) showed the greatest antioxidant activity of 7.08 ± 0.18 GAE/g extract and 75.64 ± 3.07 mg TEAC/g extract by the DPPH and ABTS assays, respectively, together with the highest FRAP value of 48.54 ± 0.48 mg FeSO₄/g extract. The antibacterial efficacy of these extracts was further evaluated against skin pathogenic bacteria, including Staphylococcus aureus, Staphylococcus epidermidis, methicillin-resistant Staphylococcus aureus (MRSA), Pseudomonas aeruginosa, Micrococcus luteus, and Cutibacterium acnes. Inhibition zones ranged from 7.00 to 14.00 mm when determined by the agar disc diffusion method. Moreover, antibacterial activity was investigated by broth dilution assay, and MIC and MBC values ranged from 0.97 to 125 mg/mL. The cytotoxicity test of propolis extracts using RAW264.7 macrophage cells showed cell viability above 70%, and the selected concentrations were used for further anti-inflammatory evaluation. T. pegdeni stingless bee propolis extracts (samples A and B) demonstrated anti-inflammatory activity by reducing NO production and decreasing the expression of iNOS, COX-2, and IL-1β genes in LPS-stimulated RAW264.7 macrophage cells. LC–MS analysis revealed differences in chemical profiles among the propolis extracts and various bioactive compounds, including phenolic acids, flavonoids, xanthones, terpenoids, triterpenoids, and curcuminoids were identified. Therefore, Thai stingless bee propolis demonstrated potential as a natural source of bioactive compounds with antioxidant, antibacterial, and anti-inflammatory properties.