Search Results (774)

Dental caries is a major global health concern, with Streptococcus mutans playing a key role in biofilm formation and acid production, which lead to enamel demineralization. Natural products, particularly plant-derived extracts, offer promising alternatives to conventional antibacterial agents. This study aimed to analyze the chemical composition of Ferula varia 70% ethanol extract (FVE) and evaluate its potential to inhibit biofilm formation by S. mutans. The aerial parts of F. varia were extracted with 70% ethanol and analyzed using LC-UV-ESI-MS/MS to determine the chemical profile. The anti-biofilm activity of FVE was evaluated using a crystal violet assay against S. mutans. Phytochemical analysis identified 14 compounds, including major phenolic acids (e.g., chlorogenic acid, gallic acid) and flavonoids (e.g., isoquercitrin, isorhamnetin-3-O-glucoside). FVE exhibited significant, dose-dependent inhibition of S. mutans biofilm formation. Importantly, the FVE concentration of 5 mg/mL inhibited S. mutans biofilm development by 100%. The potent antibiofilm activity of FVE against S. mutans is likely due to the synergistic action of its rich content of phenolic acids and flavonoids, which possess known anti-virulence properties. These findings support the potential use of FVE as a natural ingredient in oral hygiene products to prevent dental plaque and caries. Full article

Gnetol (trans-2,3′,5′,6-tetrahydroxystilbene), a naturally occurring stilbene structurally related to resveratrol (trans-3,5,4′-trihydroxystilbene; RES), has been reported to possess multiple health-promoting activities. In order to support its potential nutraceutical application, a reliable chromatography–tandem mass spectrometry (LC–MS/MS) assay was developed and validated for the quantitative determination of gnetol in mouse plasma and tissue samples, using isotopically labeled RES-¹³C₆ serving as the internal standard (IS). Electrospray ionization (ESI) was performed in negative mode, with multiple reaction monitoring (MRM) transitions m/z 243.2 → 175.0 for gnetol and m/z 233.1 → 191.0 for the IS. Chromatographic separation was achieved on a reversed-phase HPLC column using a 5-min gradient delivery of acetonitrile and 2 mM ammonium acetate at 0.5 mL/min and 40 °C. The linear calibration curve covered the concentration range of 5.0–1500 ng/mL, and the method validation confirmed its selectivity, accuracy, precision, stability, and dilution integrity. The developed method was subsequently applied to a biodistribution study in mice after oral administration of gnetol at 400 µmol/kg (equivalent to 97.7 mg/kg). Gnetol was rapidly absorbed and extensively distributed in key pharmacologically relevant organs. Despite its poor aqueous solubility, oral uptake was not significantly hindered. Collectively, these findings demonstrate that gnetol exhibits favorable absorption and tissue distribution profiles, supporting its promise as a candidate for nutraceutical development. Full article

Extracellular vesicles (EVs) are bilayer-membrane cellular components that deliver protected cargo to the extracellular environment and can mediate long-distance signaling. We have previously reported that EVs isolated from the virulent fungal pathogen Paracoccidioides brasiliensis Vpb18 can revert the expression, in the attenuated variant Apb18, of stress-related virulence traits. We presently show that the Vev and Aev, respectively, produced by these variants display distinct proteomes, with prevalent functional enrichment in Vev related to oxidative stress response, signal transduction, transport, and localization, in addition to richer protein–protein interaction. Proteome sequences were obtained by nanoflow liquid chromatography coupled with tandem mass spectrometry (nano LC-ESI-MS/MS). The Vev and corresponding Vpb18 proteomes also differed, suggesting a selective bias in vesicle protein cargo. Moreover, sublethal oxidative (VevOxi) and nitrosative (VevNO) stress modulated the Vev proteome and a positive correlation between VevOxi/VevNO-enriched and Vev-enriched (relative to Aev) proteins was observed. Out of 145 fungal virulence factors detected in Vev, 64% were enriched, strongly suggesting that molecules with virulence roles in Paracoccidioides are selectively concentrated in Vev. Our study significantly advanced the field by exploring protein N-terminal acetylation to a dimension rarely investigated in fungal EV proteomics. The proportion of N-terminally acetylated proteins in Vev was higher than in Vpb18 and the presence of Nt-acetylation in Vev-enriched virulence factors varied across the samples, suggesting that it might interfere with protein sorting into EVs and/or protein functionality. Our findings highlight the relevance of our fungal model to unraveling the significance of fungal EVs in pathogenesis and phenotypic transfer. Full article

Background/Objectives: Enhancing endothelial nitric oxide (NO) bioavailability through natural products may provide a promising strategy for the prevention and management of hypertension. This study investigated the phytochemical composition of ethanolic lotus (Nelumbo nucifera) seed extract (LSE), its vasorelaxant mechanisms, effects on endothelial NO production, and antihypertensive activity. Methods: LSE was characterized via LC-ESI-QTOF-MS using accurate mass data and fragmentation patterns. Vasorelaxant effects were evaluated in isolated rat aortas, and the underlying mechanisms were explored using pharmacological inhibitors. NO production was assessed in human endothelial EA.hy926 cells. Hypotensive activity was examined in normotensive rats following intravenous administration of LSE (10, 30, and 100 mg/kg). Molecular docking was performed to analyze interactions between LSE bioactive compounds and endothelial nitric oxide synthase (eNOS). Results: LC-ESI-QTOF-MS analysis identified 114 compounds, including primary and secondary metabolites. LSE induced vasorelaxation in endothelium-intact aortas, which was reduced by endothelium removal (p < 0.001) and by L-NAME (p < 0.001). LSE also inhibited receptor-operated, Ca²⁺ channel-mediated vasoconstriction (p < 0.05). In vivo, LSE decreased blood pressure in a dose-dependent manner. In EA.hy926 cells, LSE (750 and 1000 µg/mL) increased NO production, an effect attenuated by L-NAME. Molecular docking showed that LSE alkaloids, including nelumborine, nelumboferine, neferine, and isoliensinine had strong affinities for binding with eNOS at the tetrahydrobiopterin (BH4) binding site. Nelumborine exhibited the highest affinity, suggesting its potential as an eNOS modulator. Conclusions: LSE promotes vasorelaxation through the stimulation of endothelium-derived NO release and Ca²⁺ influx inhibition, contributing to blood pressure reduction. These findings support LSE as a potential natural antihypertensive supplement. Full article

Background/Objectives: Hypericum cerastoides (Spach) N. Robson is a lesser-known species with potential pharmacological importance. This study aimed to profile phenolic compounds in its aerial parts and assess biological activities of isolated constituents, focusing on radical-scavenging, anti-α-glucosidase, and pro-lipase effects. Methods: Phenolic compounds from H. cerastoides aerial parts were dereplicated via UHPLC-HRMS/MS. The structures of isolated compounds were determined using spectroscopic methods (1D and 2D NMR, UV, and HRMS-ESI). Radical-scavenging was evaluated by DPPH and ABTS assays; anti-α-glucosidase and pro-lipase activities were measured by LC-MS. Results: UHPLC-HRMS profiling of a hydroalcoholic extract tentatively identified and quantified 39 phenolic compounds, mainly flavonoids and hydroxycinnamic acid derivatives. Furthermore, two new phenolic compounds, namely hypercerastoside A (HC4) and hypercerastoside B (HC6), together with three known compounds, coumaroylquinic acid (HC1), myricetin-3-O-glycoside (HC2), and myricetin-3-O-galactoside (HC3), as well as two artifacts, namely methyl ester of chlorogenic acid (HC5) and hypercerastoside C (HC7), were isolated from the ethylacetate extract of the aerial parts of title plant. Compounds HC2, HC3, and HC5 displayed the highest radical-scavenging activity. The anti-α-glucosidase test showed that compounds HC1 (IC₅₀ = 44 µM) and HC3 (IC₅₀ = 206 µM) possessed similar activity to acarbose (IC₅₀ = 206 µM). Myricetin glycosides HC2 and HC3 enhanced lipase activity fivefold at 200 µM. Conclusions: H. cerastoides is a promising source of bioactive phenolic compounds with significant radical-scavenging and enzyme-modulating activities. These preliminary findings support further exploration of its therapeutic potential, especially for oxidative stress-related disorders, type 2 diabetes, and cachexia. Full article

Perovskia atriplicifolia Benth., a perennial aromatic plant widespread in Iran’s Sistan and Baluchestan region, is known for its essential oil composition, rich in aromatic and non-aromatic sesquiterpenes. To the best of our knowledge, limited information exists on the composition of its non-volatile extracts. Herein, the phytochemical investigation of the EtOH extract of P. atriplicifolia aerial parts was performed, guided by an analytical approach based on LC-(-)ESI/QExactive/MS/MS. This led to the identification of phenolics, flavonoids, diterpenes (mainly carnosic acid derivatives), and triterpenes. Structural elucidation was performed via NMR and HRMSMS analysis. Furthermore, considering the occurrence of diterpenes closely related to carnosic acid and carnosol, known for their antioxidant properties, the antioxidant activity of the extract (0.5–5.0 μg/mL) and selected pure compounds (0.5–25 μM; compounds 5, 7, 9, 10, 12, 16) was evaluated in Caco-2 intestinal cells, showing significant reduction in free radical levels. The quantitative results highlighted that the above cited compounds occurred in concentrations ranging from 1.73 to 520.21 mg/100 g aerial parts, with carnosol (12) exhibiting the highest concentration (520.21 mg/100 g aerial parts), followed by 1α-hydroxydemethylsalvicanol (9) (91.73 mg/100 g aerial parts) and carnosic acid (16) (88.16 mg/100 g aerial parts). Full article

Merlot red wines rank among the most distinguished varietals globally. This study aimed to characterize the phenolic compound profiles of Merlot wines and assess the influence of geographical origin and vintage on samples from two Albanian wine regions. Using liquid chromatography coupled with tandem mass spectrometry, a total of 31 phenolic compounds were identified and quantified. These were classified into hydroxybenzoic acids and flavan-3-ols (13 compounds), phenolic acids (9), flavonols (5), and stilbenoids (4). The total phenolic content ranged from 294 mg L⁻¹ in wines from the Mati–Mirdita region to 480 mg L⁻¹ in those from the Durrës–Kavaja region, demonstrating significant regional variation. Notably, the hydroxybenzoic acids and flavan-3-ols exhibited the most pronounced differences, with gallic acid concentrations varying from 123 mg L⁻¹ (Mati–Mirdita) to 170 mg L⁻¹ (Durrës–Kavaja). Both regions’ wines were rich in catechin, epicatechin, procyanidin derivatives, trans-caftaric acid, and ethyl gallate. However, procyanidins were found in higher concentrations in the Mati–Mirdita wines, while other phenolics were more abundant in Durrës–Kavaja samples. These findings underscore the influence of geographical and climatic factors on phenolic composition, offering a robust chemical fingerprinting approach for assessing wine authenticity and quality. Full article

Olea europaea L. ‘Lavagnina’ is cultivated in the Eastern Ligurian coast (Italy), and during the pruning process a huge amount of pruning residues is produced. This by-product is generally disposed of by burning, despite still containing bioactive compounds. In particular, olive leaves are indeed rich in secondary metabolites, which can vary both in quality and quantity in relation to the cultivar considered and the area of cultivation. For this reason, we aimed to carry out a pharmacognostic study of the pruned leaves of the unexplored local cultivar ‘Lavagnina’, evaluating the possibility of reusing this by-product for new health applications. The micromorphological characterization was conducted by light and scanning electron microscopy. ‘Lavagnina’ leaf was micromorphologically similar to that of other olive cultivars; however, it differed in terms of midrib structure. Leaf extracts were obtained using solvents of increasing polarity (petroleum ether, chloroform, methanol) and the food-grade solvent, 70% ethanol. A high antioxidant activity was found only for the methanolic (ME) and hydroalcoholic (HAE) extracts, and, therefore, they were then characterized from a phytochemical point of view by LC-ESI-HR-MS. Such analysis allowed the identification of secondary metabolites belonging mainly to secoiridoids, flavonoids, and iridoids. Overall, the HAE had the highest antioxidant activity (17.3 ± 0.6 μg/mL), and it is, therefore, the best candidate for health applications related to a protective effect on a variety of inflammation-related diseases, also considering that inflammation may play a role in cancer progression. Full article

Abeliophyllum distichum is rich in polyphenols and flavonoids with various bioactivities; however, studies on enzymatic modifications to enhance its functional properties remain limited. This study investigated the effect of Viscozyme^® L treatment on the secondary metabolite profile of A. distichum leaves. Phytochemical profiling using liquid chromatography–electrospray ionization tandem mass spectrometry revealed a decrease in the total number of detectable compounds, from 26 in the untreated extract to 16 in the enzyme-treated extract. Following Viscozyme^® L treatment, a notable shift in metabolite composition was observed, with significant enrichment of flavonoid glycosides, pyranone derivatives, and amino acid-related metabolites. Quantitative high-performance liquid chromatography analysis showed significant reductions in glycosylated compounds such as rutin (1), acteoside (2), and isoacteoside (3), while the aglycone quercetin (4) content increased more than four-fold compared to the control. These results indicate that Viscozyme^® L facilitates the deglycosylation of flavonoid glycosides into their aglycone forms. This enzymatic transformation suggests a potential strategy to enhance the bioavailability and functional value of A. distichum leaf extracts for nutraceutical and pharmaceutical applications. Full article

Glycolipids are structurally diverse amphiphilic molecules with potential as non-petrochemical-derived bioproducts, including surfactants, emulsifiers, and antioxidants. The different bioactivities associated with this range of glycolipid structures also present opportunities for dietary supplements, cosmetics, and pharmaceuticals. Marine glycolipids are underexplored due to challenges with purification and structural characterisation. Analytical approaches enabling efficient sample purification, isolation, and identification of target glycolipids are crucial to determining the bioactivity and functions of organisms such as shellfish and seaweed. This review summarises advances in analytical methods applicable to marine glycolipids, including extraction and enrichment methods tailored to specific subclasses. Thin-layer chromatography (TLC)-based rapid detection techniques developed for specific subclasses in complex biological samples are discussed, alongside structure identification methods based on liquid chromatography (LC)–electrospray ionisation (ESI)–tandem mass spectrometry (MS/MS). Hydrophilic interaction liquid chromatography (HILIC), reverse-phase liquid chromatography (RPLC), and supercritical fluid chromatography (SFC) coupled with MS detection are reviewed for their application to glycolipids. The application of two-dimensional liquid chromatography (2D-LC) and advanced MS-based approaches that facilitate both the rapid resolution and comprehensive characterisation of molecular species are also reviewed. Full article

Background: Nephrolepis exaltata (sword fern) possesses a considerable amount of phytochemicals and different biological activities. The current study investigates the anti-biofilm potential of greenly synthesized bimetallic nanoparticles of Nephrolepis exaltata leaf methanol extract (NEME-MnO₂-MgO BNPs). Methods: The NEME was subjected to UPLC/MS analysis, followed by characterization of its NPs by size, zeta potential, FTIR, entrapment efficiency, and release. Then, antioxidant, antimicrobial and antibiofilm assays were employed, followed by in silico studies. Results: The UPLC/MS analysis of NEME led to the tentative identification of 27 metabolites, mostly phenolics. The MnO₂-MgO BNPs presented a uniform size and distribution and exhibited IC₅₀ values of 350 and 215.6 μg/mL, in the DPPH and ABTS assays, respectively. Moreover, the NPs exhibited antimicrobial and anti-biofilm efficacies against Pseudomonas aeruginosa, Klebsiella pneumonia (ATCC-9633), Staphylococcus aureus (ATCC-6538), Escherichia coli, Bacillus cereus, and C. albicans, with MIC values of 250–500 μg/mL. The MnO₂-MgO BNPs inhibited Candida albicans biofilms with a % inhibition of 66.83 ± 2.45% at 1/2 MIC. The network pharmacology highlighted epigallocatechin and hyperoside to be the major compounds responsible for the anti-biofilm potential. The ASKCOS facilitated the prediction of the redox transformations that occurred in the green synthesis, while the docking analysis revealed enhanced binding affinities of the oxidized forms of both compounds towards the outer membrane porin OprD of P. aeruginosa, with binding scores of −4.6547 and −5.7701 kcal/mol., respectively. Conclusions: The greenly synthesized Nephrolepis exaltata bimetallic nanoparticles may provide a promising, eco-friendly, and sustainable source for antimicrobial agents of natural origin with potential biofilm inhibition. Full article

Rhododendron tomentosum Harmaja is a marsh plant known for its high content of bioactive components, including essential oil, flavonoids, and phenolic acids. In the current work, the effects of cultivation mode (agar, liquid stationary, shake flask, and temporary immersion) and experiment duration (30, 60, and 90 days) on the growth and contents of non-volatile phenolics in Rhododendron tomentosum microshoots were investigated. Agar and liquid stationary cultures provided the highest dry biomass yield per liter, but their dry weight productivities per day were the lowest among the tested systems. Agitated and temporary immersion cultures, on the other hand, were the most productive in terms of fresh and dry biomass yield per day. LC-DAD-ESI-MS analysis of extracts from microshoots and wild-grown plants revealed similarities in phenolic composition: in both cases, the presence of catechin, chlorogenic acid, and flavonoids of flavonol type (mainly glycosides of quercetin and myricetin) was confirmed. The qualitative composition of the phenolic fraction was not affected by experiment duration and cultivation mode. As determined by HPLC analysis, shake flask and temporary immersion cultures were characterized by the highest phenolic contents: up to 37.5 and 26 mg/g dry weight, respectively. The maximum productivities of the above systems were equal to 18 and 13.5 mg/L/d, respectively. Full article

Background/Objectives: Vinegar–egg is a traditional health-promoting beverage prepared by soaking eggs in vinegar. While both eggs and vinegar are common dietary components with well-documented nutritional and pharmacological activities, eggs treated with vinegar have been rarely studied. This study aims to identify and characterize bioactive compounds in vinegar–egg and investigate their potential wound-healing activities. Methods: The vinegar–egg extract was analyzed using liquid chromatography–mass spectrometry (LC–MS) and column chromatography, including HPLC purification, which led to the isolation of four phenolic compounds. Results: These compounds were identified as 4-hydroxybenzoic acid (1), vanillic acid (2), methyl syringate (3), and leptosperin (4) using ESI-MS, UV, and NMR spectroscopic data. Among the isolates, 4-hydroxybenzoic acid (1) and vanillic acid (2) demonstrated wound-healing properties in mouse embryonic fibroblast (MEF) cells. None of the compounds, 4-hydroxybenzoic acid (1), vanillic acid (2), methyl syringate (3), or leptosperin (4), exhibited cytotoxicity in PC12, AGS, MEF, or MDA-MB-231 cells. Notably, 4-hydroxybenzoic acid (1) enhanced cell motility by 2.59-fold and cell invasion by 1.20-fold, while vanillic acid (2) increased cell motility by 2.69-fold and cell invasion by 1.23-fold. Western blot analysis revealed that treatment with 4-hydroxybenzoic acid (1) and vanillic acid (2) increased the phosphorylation of focal adhesion kinase (p-FAK) and matrix metalloproteinase 2 (MMP-2). Furthermore, both compounds elevated the phosphorylation of p38, a key regulator in wound-healing pathways. Conclusions: These findings demonstrate that 4-hydroxybenzoic acid (1) and vanillic acid (2) accelerate wound healing through the activation of focal adhesion and mitogen-activated protein kinase (MAPK) signaling pathways. These results highlight vinegar–egg as a promising therapeutic candidate for wound healing. Full article

Chemical products, including cleaning agents, disinfectants, stain removers, and cosmetics, release harmful chemicals that pose a risk to human health and the environment, necessitating alternative sources. The objective of this research was to identify the most effective phytoextract from food production waste for use in sustainable aerosol hygiene technology as an electrostatic bio-disinfectant. The investigation was performed through wipe tests and airborne microbial collection techniques. The upgraded coffee silverskin phytoextract demonstrated superior disinfection potential for various surfaces and airborne microbes compared to the hot trub phytoextract, with an industrial disinfectant serving as the control. Log reduction analyses revealed a more significant killing efficacy (p ≤ 0.05, using the ANOVA test) against Gram-positive organisms (Bacillus subtilis and Listeria monocytogenes) than against Gram-negative organisms (Escherichia coli and Vibrio parahaemolyticus), with the log reductions ranging from 3.08 to 5.56 and 3.72 to 5.81, respectively. Chemical characterization by LC-ESI-QTOF-MS, ¹H NMR, and FTIR showed that CGAs and chalcones are the most bioactive compounds in CSS and HT, respectively. The innovation in this work involves an integrated approach that combines waste-derived phytoextracts, advanced chemical profiling, and scalable aerosol disinfection. Furthermore, this research offers a greener, cost-effective, and industrially relevant alternative to synthetic chemical disinfectants. The interdisciplinary approach contributes to the development of bio-based disinfectants for use in the food industry, hospitals, and public health settings. This investigation supports a paradigm shift toward sustainable disinfection practices, thereby improving food and environmental safety. Full article

Rouxiella badensis DSM 100043^T had been previously proven to produce a novel glucoselipid biosurfactant which has a very low critical micelle concentration (CMC) as well as very good stability against a wide range of pH, temperature, and salinity. In this study, we performed a function-based library screening from a R. badensis DSM 100043^T genome library to identify responsible genes for biosynthesis of this glucoselipid. The identified open reading frames (ORFs) were cloned into several constructs in Escherichia coli for gene permutation analysis and the individual products were analyzed using high-performance thin-layer chromatography (HPTLC). Products of interest from positive expression strains were purified and analyzed by liquid chromatography/electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) and nuclear magnetic resonance (NMR) for further structure elucidation. Function-based screening of 5400 clones led to the identification of an operon containing three ORFs encoding acetyltransferase GlcA (ORF1), acyltransferase GlcB (ORF2), and phosphatase/HAD GlcC (ORF3). E. coli pCAT2, with all three ORFs, resulted in the production of identical R. badensis DSM 100043^T glucosedilipid with Glu-C_10:0-C_12:1 as the main congener. ORF2-deletion strain E. coli pAFP1 primarily produced glucosemonolipids, with Glu-C_10:0,3OH and Glu-C_12:0 as the major congeners, predominantly esterified at the C-2 position of the glucose moiety. Furthermore, fed-batch bioreactor cultivation of E. coli pCAT2 using glucose as the carbon source yielded a maximum glucosedilipid titer of 2.34 g/L after 25 h of fermentation, which is 55-fold higher than that produced by batch cultivation of R. badensis DSM 100043^T in the previous study. Full article