Search Results (50)

Safflower (Carthamus tinctorius L.) is a cash crop grown worldwide. Its seeds and flowers are primarily processed for edible oil and medicinal raw materials, while stems and leaves are discarded as agricultural waste. Recently, plant-derived exosome-like nanoparticles (PELNs) have gained growing research interest in food nutrition owing to their exceptional biocompatibility and relatively low cost for large-scale production. This study aimed to explore the potential functional value of safflower leaf agricultural waste by isolating safflower-derived PELNs. Firstly, a protocol was established for the isolation and purification of safflower (Carthamus tinctorius L.)-derived exosome-like nanoparticles (Ct-ELNs) from safflower leaves using sucrose density gradient ultracentrifugation. NTA analysis revealed that the 30–45% sucrose fraction enriched with Ct-ELNs exhibited the most uniform particle size, highest particle concentration, and optimal purity. Transmission electron microscopy confirmed typical PELNs ultrastructure: disc- or cup-shaped vesicles surrounded by bilayer lipid membranes. FM4-64 fluorescence suggests time-dependent association and likely cellular uptake of labeled Ct-ELNs by IPEC-J2 cells. Cellular assays demonstrated that Ct-ELNs elevated IPEC-J2 cell metabolic activity under matched protein-normalized dosing conditions. The 30–45% sucrose fraction showed the most favorable physicochemical profile and preliminary in vitro protective effects, including improved IPEC-J2 cell metabolic activity and modulation of Enterotoxigenic Escherichia coli (ETEC)-induced inflammation-related gene expression. Overall, this study established an optimized isolation protocol for Ct-ELNs derived from safflower leaves. These data indicate that safflower-derived Ct-ELNs confer preliminary cytoprotective transcriptional regulatory effects on intestinal epithelial cells under in vitro culture conditions. Full article

Fruit tree leaves are an abundant agro-waste material with promising yet underexplored biological potential. This study compared the biological activity of aqueous extracts obtained from apple (Malus domestica), apricot (Prunus armeniaca), and cherry (Prunus cerasus) leaves and their kombucha-fermented counterparts in the context of cosmetic and dermatological applications. Phytochemical composition before and after fermentation was analyzed chromatographically. Antioxidant activity was evaluated using DPPH, ABTS, and FRAP assays, while intracellular reactive oxygen species (ROS) levels in keratinocytes and fibroblasts were assessed using the H₂DCFDA probe. Cytotoxicity was determined by Alamar Blue and Neutral Red assays. Antimicrobial activity against seven bacterial strains was investigated using minimum inhibitory concentration and disc diffusion methods. Anti-inflammatory activity was evaluated in LPS-stimulated THP-1 cells by measuring TNF-α, IL-1β, and IL-6 levels using ELISA. The influence of the samples on collagenase, elastase, and hyaluronidase activity was also analyzed. Fermentation increased the content of selected phenolic compounds and enhanced antioxidant, antimicrobial, anti-inflammatory, and anti-ageing properties. Ferments more effectively reduced oxidative stress in skin cells and showed no cytotoxicity within the tested concentration range. These findings indicate that kombucha fermentation may support the valorization of fruit tree leaf agro-waste as multifunctional ingredients for skincare formulations. Full article

Rapid alkalinization factor (RALF) peptides are recognized as multifunctional regulators of plant stress responses, yet their roles in woody species remain poorly defined. Here, we identified a RALF22-like peptide from poplar ‘Shanxin’ (Populus davidiana × P. bolleana; PdbRALF22-like) and investigated its roles in salt tolerance and disease resistance. Synthetic PdbRALF22-like peptide elicited a rapid ROS burst in poplar leaf discs. In Nicotiana benthamiana, which was otherwise unresponsive to the peptide, transient expression of either of two poplar FERONIA-like receptor kinases (PdbFER-like-1 and PdbFER-like-2) enabled peptide-triggered ROS production, consistent with receptor-matched responsiveness in a heterologous context. Using CRISPR/Cas9, we generated a PdbRALF22-like knockout line and assessed salt tolerance in vitro and soil-grown assays. Under salinity, the mutant showed sustained rooting at high NaCl concentrations and improved growth relative to wild type. After 0.2 M NaCl treatment, soil-grown mutant plants exhibited reduced wilting and leaf injury. Evans Blue, DAB, and NBT staining indicated reduced membrane damage and lower accumulation of hydrogen peroxide and superoxide in the mutant. Significantly, the same knockout line displayed increased susceptibility to infection by the poplar leaf spot fungus, with larger lesions and higher pathogen biomass, accompanied by reduced ROS output and lower induction of the defense marker gene PdbPR1. Collectively, PdbRALF22-like negatively regulates salt tolerance while contributing positively to disease resistance, and represents a regulatory node linking salinity tolerance and disease susceptibility in poplar ‘Shanxin’, with poplar FER-like receptors providing a plausible route for peptide-triggered ROS signaling. This work expands our understanding of RALF peptide signaling in woody plants. Full article

Vernonia amygdalina Delile (Asteraceae), commonly known as bitter leaf, is a tropical shrub that may potentially serve as a biopesticide against the Colorado potato beetle Leptinotarsa decemlineata (Say) (Coleoptera: Chrysomelidae), a key pest of potatoes. The beetle’s behavioral response to the methanolic leaf extract of V. amygdalina was evaluated in this study. Using no-choice, dual-choice, and wind-tunnel assays under laboratory conditions, we evaluated responses of larvae and adults to potato leaf discs treated with V. amygdalina extract in a randomized complete block design, measuring feeding behavior, leaf damage levels, and remaining leaf area. The results showed that V. amygdalina had no biocidal effects against the beetle, as no mortality was incurred. However, dose-linked antifeedant effects were evident in both no-choice and dual-choice arenas. Vernonia amygdalina minimized leaf-area loss most effectively at the highest concentration, especially against the larvae. The extract showed no olfactory repellency but acted as an antifeedant, possibly through contact or taste (gustatory) receptors. The consistent behavioral avoidance at higher concentrations suggests that V. amygdalina acts as a form of deterrent against the Colorado potato beetle. Full article

The growing emphasis on sustainability, regional distinctiveness, and spontaneous fermentation in winemaking necessitates a more comprehensive understanding of local yeast populations and their functional mechanisms. In total, 115 indigenous yeast strains were examined for their enzymatic activities of potential vitivinicultural significance. The yeasts were screened for chitinase activity (biocontrol potential), glycosidase activity (terpene release), β-lyases (thiol release), and sulfite reductases (off-flavor formation), followed by quantitative analysis of the selected subsets. Yeasts were further evaluated for inhibition of fungal mycelial growth, VOC-mediated inhibition, and tolerance to commonly applied fungicides. Pre-field selection was refined using the niche overlap index and grapevine leaf disc assay. The results confirmed chitinolytic activity in four species; all strains exhibited hydrolase activities, with H. uvarum 116 displaying the highest cell-associated activity (6.32 U/g), while T. delbrueckii Sut94 showed the highest extracellular activity (1.36 U/g). β-glucosidase and β-lyase activities were widespread, whereas hydrogen sulfide production was infrequent. P. guilliermondii ZIM 624 showed the most comprehensive overall enzymatic profile, together with strong inhibition patterns. A field trial on Pinot cultivars (V. vinifera L.) further evaluated P. guilliermondii ZIM 624 within an integrated disease management approach, with responses being more pronounced in ‘Pinot noir’ than in ‘Pinot gris’. Full article

Background: Tribulus terrestris is a medicinal plant used in traditional medicine to treat certain illnesses. Though past efforts mostly focused on the fruits and roots, current research examined the phytochemical composition and bioactivity of leaf extract (LE) and seed extract (SE). Methods: GC-MS compared phytochemical profiles, and total phenolic and flavonoid content were determined. The extracts were tested for antibacterial activity (disc diffusion, MIC/MBC), antioxidant potential (DPPH, ABTS⁺), cytotoxicity (MTT assay in MCF-7 and HepG2 cells), and anti-diabetic activity (α-amylase and α-glucosidase inhibition). Expression of apoptotic genes was also investigated. Results: The LE had a superior phytochemical composition, with greater phenolic and flavonoid levels. Compared to SE, it exhibited considerably higher antibacterial activity (MIC = 6.25–25 μg/mL), antioxidant potential (IC₅₀ = 90.71–113.41 μg/mL), cytotoxicity (IC₅₀ = 105.12–126.14 μg/mL), and enzyme inhibition (IC₅₀ = 84–96.62 μg/mL). The LE also drastically reduced the expression of anti-apoptotic genes Bcl-2 and Bcl-xL in cancer cells. T. terrestris LE has significantly higher bioactive potential than SE in a range of pharmacological arenas due to its superior phytochemically complete profile. Conclusions: The findings indicate the LE as a promising candidate for the development of standardized phytotherapeutically active compounds. Full article

In 2019, bacterial canker caused by Pseudomonas syringae pv. actinidiae was first identified in Actinidia arguta. This disease has led to significant yield reduction, plant mortality, and substantial economic losses in A. arguta cultivation. Its emergence poses a novel challenge to the sustainable global production of kiwifruit. Currently available treatments for bacterial canker caused by P. syringae pv. actinidiae are scarce. Moreover, the environmental toxicity of copper-based compounds and emerging antibiotic resistance issues necessitate the development of eco-friendly control strategies. Disease management strategies based on biocontrol bacteria have shown broad application prospects. In this study, the isolate CHHM-1 with significant antagonistic activity against P. syringae pv. actinidiae was isolated from the rhizosphere soil of healthy A. arguta. It was identified as Pseudomonas koreensis through 16S rRNA gene and whole-genome sequencing. Genomic analysis revealed that the isolate CHHM-1 harbors various genes related to biocontrol, plant growth promotion, and antibiotic resistance, suggesting strong environmental adaptability and functional potential. Furthermore, the strain exhibited multiple plant growth-promoting traits, such as nitrogen fixation, phosphate solubilization, siderophore production, and synthesis of indole-3-acetic acid (IAA). In vitro antagonism assays confirmed the strong antagonistic activity of the isolate CHHM-1 against P. syringae pv. actinidiae. A dual-culture plate assay showed an average inhibition zone of 4.36 cm, while preventive application on plants significantly reduced lesion length to 1.3 mm (vs. 6.2 mm control) in shoots and lesion area to 10% (vs. 80% control) in leaf discs. Further antibacterial tests revealed that its inhibitory mechanism is attributed to secreted antimicrobial substances. These findings provide a promising candidate for developing novel biopesticides to combat P. syringae pv. actinidiae variants, reduce chemical dependency, and foster sustainable A. arguta production. Full article

There is insufficient evidence regarding the cytotoxicity of restorative 3D-printing resins, used as part of the digital workflow in dentistry. This study presents a novel comparative evaluation of cell viability and adhesion using human Wharton’s jelly-derived mesenchymal stem cells (WJ-MSCs), a less commonly used but clinically relevant cell line in dental biomaterials research. The aim of this study was to evaluate the cell viability of WJ-MSCs seeded on 3D-printed resins intended for temporary restorations. Resin discs of three commercial 3D-printing resins (NextDent C&B, Leaf Dental C&B, and UNIZ Temp) and a conventional self-curing acrylic resin (NicTone) were used. WJ-MSCs were cultured on the specimens for 1, 4, and 10 days. Cell viability was assessed using the PrestoBlue assay, Live/Dead immunofluorescence staining, and 7AAD/Annexin V staining. Cell adhesion was evaluated using scanning electron microscopy. Direct exposure to the 3D-printed resins and the self-curing acrylic caused slight reductions in cell viability compared to the control group in both microscopic analyses. 7AAD/Annexin V showed the highest percentage of viable WBCs for the conventional acrylic (34%), followed by UNIZ (35%), NextDent (42%), and Leaf Dental (36%) (ANOVA p < 0.05 Tukey’s post-hoc test p < 0.05). These findings suggest that 3D-printed resins could be considered safe for use in temporary restorations. Full article

Background/Objectives: Bacterial contamination leads to foodborne illnesses, and new antibiotics are required to combat these pathogens. Interest has increased in medicinal plants as targets for new antibiotics. Methods: This study evaluated the antibacterial activity of leaf extracts from Terminalia citrina (Gaertn.) Roxb. ex Fleming against four bacterial pathogens (including a methicillin-resistant Staphylococcus aureus (MRSA) strain) using disc diffusion and liquid microdilution assays. The phytochemical composition of the extracts were determined using ultra-high-performance liquid chromatography–mass spectrometry (UPLC-MS). Results: Both the aqueous and methanol extracts demonstrated noteworthy antibacterial activity against Bacillus cereus (MICs of 468.8 µg/mL and 562.5 µg/mL, respectively). Additionally, the extracts were effective against MRSA (MICs = 625 µg/mL). Strong antibacterial effects were also observed against S. aureus, with MICs of 625 µg/mL (aqueous extract) and 833.3 µg/mL (methanol extract). Twelve combinations of extracts and conventional antibiotics were synergistic against B. cereus and S. flexneri. UPLC-MS analysis revealed two flavonoids, orientin 2″-O-gallate and astragalin, exclusive to the aqueous extract, whilst pinocembrin and gallic acid were only detected in the methanol extract. Both extracts contained vitexin 2″-O-p-coumarate, ellagic acid, orientin, rutin, chebulic acid, terminalin, and quercetin-3β-D-glucoside. Both extracts were determined to be nontoxic. Conclusions: The abundance and diversity of polyphenols in the extracts may contribute to their strong antibacterial properties. Further research is required to investigate the antibacterial effects of the individual extract compounds, including their effects when combined with conventional antibiotics, and the potential mechanisms of action against foodborne pathogens. Full article

The increasing resistance of microorganisms to currently used antimicrobials requires the urgent development of new effective treatments. Plant-based natural products can be an alternative solution. The aerial plant parts of the cranberry (Vaccinium macrocarpon) present a potential new source of antimicrobial secondary metabolites. Volatile essential oils were extracted from Stevens, Early Black, and Mullica Queen variety plants by steam distillation (SD) and the Clevenger method (CM), and their profiles were characterized by GC-MS. The extracts and two identified constituents, cinnamaldehyde and terpineol, were screened by the disc diffusion assay against Gram-positive B. cereus ATCC 11778 and S. aureus ATCC 25923 and Gram-negative bacteria E. coli ATCC 25922, P. aeruginosa ATCC 27853, and C. albicans ATCC 14053. Radical scavenging antioxidant activity was also determined using the DPPH assay. The CM extracts were rich in fatty acids, sesquiterpenes, and diterpenes, whereas the SD extracts contained more aldehydes, monoterpenes, and phenylpropanoids. All volatile extracts showed promising antioxidant activity; leaf extract activity was significantly higher than the vine (p < 0.05). The CM leaf and vine extracts exhibited antimicrobial activity against B. cereus, S. aureus, E. coli, and C. albicans compared to the SD, and the leaf extracts were more effective than the vine extracts. Individual constituents of leaf and vine extracts, cinnamaldehyde and α-terpineol, also showed antimicrobial activity against these organisms. The active constituents of the CM extracts are yet to be identified. A multivariate analysis revealed a particular pattern of inhibition of the tested organisms. Based on our results, cranberry volatile extracts have potential for future valorization as antibacterials, antifungals, and antioxidants. Full article

Urtica dioica L. has been used as a natural remedy due to its healing properties for over 2000 years. The aim of this study is to investigate the chemical composition, antimicrobial, antioxidant, and antitumor properties in vitro of the aqueous extract of Urtica dioica leaves (AEUD). The chemical composition was assessed by an ultra-high-performance liquid chromatography system coupled to a benchtop QExactive high-resolution accurate mass spectrometry operating in a data-dependent acquisition mode as a non-target approach. Minimal inhibitory concentration (MIC) and disc diffusion were used to assess the antibacterial efficacy against nine bacterial strains. The antioxidant impact was assessed using DPPH, ABTS, FRAP, and ferrous ion-chelating ability assays. By using the MTT method, the cytotoxicity effect of AEUD on colon cancer cell HCT-116 was evaluated. Flow cytometry was used to analyze the cell cycle. Finally, the anti-migration and anti-invasion properties of AEUD on HCT-116 cells were estimated using the wound healing test and Transwell assays. AEUD is a rich source of phenolic compounds. The results of disc diffusion and MIC showed that the AEUD is more active against Gram-positive bacteria than against Gram-negative bacteria. MTT assay confirmed that the AEUD inhibited HCT-116 colon cancer cell proliferation. Findings of flow cytometry confirmed that cell cycle arrest occurred at the G2 phase. Additionally, AEUD had anti-migration and anti-invasion effects. This study shows that Urtica dioica aqueous leaf extract exhibits potential antibacterial, antioxidant, and antitumoral activities on HCT-116 colon cancer cells. Full article

Elevated salinity negatively impacts plant growth and yield, presenting substantial challenges to agricultural and forestry productivity. The bHLH transcription factor family is vital for plants to cope with various abiotic stresses. However, it remains uncertain whether bHLH transcription factors can regulate salt stress in Populus ussuriensis. In the following study, a salt-induced bHLH transcription factor PubHLH66 was identified from P. ussuriensis. PubHLH66 has a typical and conserved bHLH domain. Subcellular localization and yeast two-hybrid (Y2H) assays confirmed that it is a nucleus-localized transactivator and the activation region is located at the N-terminus. PubHLH66-OE and PubHLH66-SRDX transgenic P. ussuriensis were obtained through Agrobacterium-mediated leaf disc transformation. Morphological and physiological results demonstrated that PubHLH66-OE enhanced salinity tolerance, as indicated by reduced electrolyte leakage (EL), malondialdehyde (MDA), and H₂O₂ levels, along with increased proline contents and activities of peroxidase (POD) and superoxide dismutase (SOD). In contrast, PuHLH66-SRDX poplar showed decreased salt tolerance. Quantitative real-time PCR (RT-qPCR) confirmed that PubHLH66 enhanced salt tolerance by regulating the expression of genes such as PuSOD, PuPOD, and PuP5CS, resulting in reduced reactive oxygen species (ROS) accumulation and an improved osmotic potential. Thus, PubHLH66 could be a candidate gene for molecular breeding to enhance salt tolerance in plants. These results laid a foundation for exploring the mechanisms of salt tolerance in P. ussuriensis, facilitating the development of more salt-tolerant trees to combat the increasing issue of soil salinization globally. Full article

Biofilms are bacterial communities on surfaces within an extracellular matrix. Targeting biofilm-specific bacteria is crucial, and natural compounds with reported antibiofilm activity have garnered significant interest. The study evaluated the antibacterial and antibiofilm activity of Erythrina senegalensis leaf extract against multidrug-resistant (MDR) Gram-negative bacteria, including Salmonella Typhimurium, S. Typhi, S. Enteritidis, Klebsiella pneumoniae, and Pseudomonas aeruginosa. The leaf extract was prepared using aqueous and ethanol solvents, and qualitative phytochemical screening revealed the presence of various bioactive compounds such as tannins, saponins, cardiac glycosides, flavonoids, terpenoids, alkaloids, anthraquinone, reducing sugar, and ketones. A Kirby–Bauer disc diffusion assay was performed to test the susceptibility of antibiotics, and the antibacterial efficacy of the aqueous and ethanol extracts of E. senegalensis was determined using the cup-plate method, while the antibiofilm activities were determined using the crystal violet titer-plate method. The aqueous and ethanol extracts of E. senegalensis revealed the presence of tannins, saponins, cardiac glycosides, flavonoids, terpenoids, alkaloids, anthraquinone, reducing sugar, and ketones. The study found that the Gram-negative bacteria isolates that were MDR were S. Typhimurium, S. Enteritidis, and P. aeruginosa, while K. pneumoniae was resistant to beta-lactam and fluoroquinolones, and S. Typhi was susceptible to all antibiotics tested. Statistically, susceptibility to antibiotics had an inverse, weak, and significant relationship with biofilm production (r = −0.453, −0.106, −0.124, −0.106, −0.018, n = 10, p < 0.05). The aqueous extract showed good biofilm inhibition against K. pneumoniae and P. aeruginosa, and poor biofilm inhibition against S. Enteritidis, while S. Typhimurium and S. Typhi exhibited no biofilm inhibition. The ethanol extract did not demonstrate any antibiofilm activity against the tested Gram-negative pathogens. The study suggests that the Gram-negative bacteria’s capacity to form biofilms is negatively associated with their antibiotic resistance phenotypes, and the aqueous extract of E. senegalensis exhibited moderate antibiofilm activity against K. pneumoniae, P. aeruginosa, and S. Enteritidis. Full article

Assessing salt tolerance in plants under field conditions is a challenging task. The objective of this research was to assess the effectiveness of different methods (leaf disc assay and pot experiment) for evaluating salt tolerance in rice. Using two varieties with different salt tolerance, Changmaogu (CM) and 9311, under three NaCl levels (0, 0.3%, and 1.0%), we evaluated the photosynthetic performance in terms of chlorophyll content in leaf disc assays, as well as the photosynthetic rate (Pn), chlorophyll content, linear electron flow (LEF), and non-photochemical quenching (NPQ), in a semi-controlled pot experiment. In the leaf disc assay, CM showed a smaller decrease in chlorophyll content compared to 9311, especially under 1.0% salinity. Simultaneously, in the pot experiment, the CM variety employed flexible photosynthetic strategies, actively decreasing LEF and Pn after 5 days of salt stress (day 5) and then increasing photosynthetic capacity (chlorophyll content, LEF, and Pn) on day 10. Notably, the total chlorophyll content for the CM variety under 1.0% salinity was significantly higher than in the control, showing a 25.0% increase. Additionally, CM demonstrated NPQt sensitivity under 0.3% salinity, requiring an LEF of 150 to achieve an NPQt value of 3.0, compared to an LEF of 180 in the control. These results suggest that a simple leaf disc assay may not fully capture the adaptive mechanisms of rice plants under salinity stress. Therefore, we advocate for the use of more comprehensive methods, such as outdoor pot or field experiments, to gain a deeper understanding and more accurate evaluation of salt tolerance in rice. Full article

Background: Plant-derived products or extracts are widely used in folk/traditional medicine to treat several infections, ailments, or disorders. A well-known medicinal herb, Myrtus communis is an evergreen fragrant plant native to the Mediterranean region that has been used for ages in traditional medicine around the world. Materials and methods: The microplate alamarBlue assay and the well diffusion method were used to evaluate the zone of inhibition and MIC, respectively. The double-disc diffusion method was used to investigate the synergy between antibiotics and the extract. The crystal violet method was used to investigate biofilm development. The SulphoRhodamine-B assay and DNA flow cytometry were used to investigate the proliferation and subsequent distribution of cells among different phases of the cell cycle. The apoptotic and necrotic phases of the cancer cells were examined using flow cytometry in conjunction with Annexin V-FITC/PI labeling. Using the IBM SPSS statistical program, a one-way ANOVA with Tukey’s post hoc test was employed for statistical analysis. Results: The ethanolic leaf extract of M. communis showed a strong growth inhibition effect (zone of inhibition: 20.3 ± 1.1–26.3 ± 2.5 mm, MIC: 4.88–312.5 µg/mL, and MBC: 39.07–1250 μg/mL) against several rapidly growing and slow-growing mycobacterial strains in a dose-dependent manner. Damage to the cell wall of bacterial cells was determined to be the cause of the antimycobacterial action. The extract inhibited biofilm formation (MBIC of 9.7 µg/mL) and eradicated already-formed mature and ultra-mature biofilms of M. smegmatis, with MBEC values of 78 µg/mL and 156 µg/mL, respectively. Additionally, the extract exhibited potent anticancer effects against diverse cancer cell lines of the breast (MCF-7), liver (HepG2), cervix (HeLa), and colon (HCT116) (IC₅₀ for HCT116: 83 ± 2.5, HepG2: 53.3 ± 0.6, MCF-7: 41.5 ± 0.6, and HeLa: 33.3 ± 3.6) by apoptosis after arresting the cells in the G₁ phase of the cell cycle. Conclusions: These results suggest that M. communis leaf extract is a potential source of secondary metabolites that could be further developed as potential anticancer and antimycobacterial agents to treat diverse types of cancers and mycobacterial infections. Full article