Search Results (152)

Background/Objectives: Senecio scandens Buch.-Ham. is a flavonoid-rich traditional medicinal plant with established immunomodulatory properties. However, the mechanisms underlying the immunoregulatory and intestinal protective effects of its flavonoid extract (Senecio scandens flavonoids—SSF) remain unclear. This study characterized SSF’s bioactive components and evaluated its efficacy against cyclophosphamide (CTX)-induced immunosuppression and intestinal injury. Methods: The constituents of SSF were identified using UHPLC/Q-Orbitrap/HRMS. Mice with CTX-induced immunosuppression were treated with SSF (80, 160, 320 mg/kg) for seven days. Immune parameters (organ indices, lymphocyte proliferation, cytokine, and immunoglobulin levels) and gut barrier integrity markers (ZO-1, Occludin, Claudin-1 protein expression; sIgA secretion; microbiota composition) were assessed. Network pharmacology combined with functional assays elucidated the underlying regulatory mechanisms. Results: Twenty flavonoids were identified in SSF, with six prototype compounds detectable in the blood. The SSF treatment significantly ameliorated CTX-induced weight loss and atrophy of the thymus and spleen. It enhanced splenic T- and B-lymphocyte proliferation by 43.6% and 29.7%, respectively; normalized the CD4⁺/CD8⁺ ratio (1.57-fold increase); and elevated levels of IL-2, IL-6, IL-10, TNF-α, IFN-γ, IgM, and IgG. Moreover, SSF reinforced the intestinal barrier by upregulating tight junction protein expression and sIgA levels while modulating the gut microbiota, enriching beneficial taxa (e.g., the Lachnospiraceae_NK4A136_group, Akkermansia) and suppressing pathogenic Alistipes. Mechanistically, SSF activated the TLR/MyD88/NF-κB pathway, with isoquercitrin identified as a pivotal bioactive constituent. Conclusions: SSF effectively mitigates CTX-induced immunosuppression and intestinal damage. These findings highlight SSF’s potential as a dual-functional natural agent for immunomodulation and intestinal protection. Subsequent research should validate isoquercitrin’s molecular targets and assess SSF’s clinical efficacy. Full article

Arugula leaves (Diplotaxis tenuifolia L. and Eruca sativa L.) are a must-have ingredient in ready-to-eat salads, as they are prized for their appearance, taste, and flavor. The nutraceutical properties of this leafy vegetable are attributed to the presence of valuable secondary metabolites, such as phenolic acids and glucosinolates. Using UHPLC-Q-Orbitrap HRMS analysis and ion chromatography, we characterized the content of phenolic acids, glucosinolates, nitrates, and organic acids in organic arugula [Diplotaxis tenuifolia (L.) DC] and evaluated how the foliar application of three different non-microbial biostimulants (a seaweed extract, a vegetable protein hydrolysate, and a tropical plant extract) modulated the expression of these. Although the application of vegetable protein hydrolysate increased, compared to control plants, the nitrate content, the application of the same biostimulant increased the total content of glucosinolates and phenolic acid derivatives by 5.2 and 17.2%. Specifically, the foliar application of the plant-based biostimulant hydrolyzed protein significantly increased the content of glucoerucin (+22.9%), glucocheirolin (+76.8%), and ferulic acid (+94.1%). The highest values of flavonoid derivatives (173.03 μg g⁻¹ dw) were recorded from plants subjected to the exogenous application of seaweed extract. The results obtained underscore how biostimulants, depending on their origin and composition, can be exploited not only to improve agronomic performance but also to enhance the nutraceutical content of vegetables, guaranteeing end consumers a product with premium quality characteristics. Full article

Asparagus (Asparagus officinalis L.) is widely recognized for its nutritional and functional properties, attributed to its rich content of polyphenols and antioxidant compounds. However, the content of compounds that remains bioaccessible following typical domestic preparation and digestion remains unclear. This study assessed the polyphenolic profile and antioxidant capacity of the edible portion of two A. officinalis cultivars (Placoseps and Darlise), harvested in different seasons, in edible form, cooked (using boiling on an induction cooktop), and cooked-digested extracts. Rutin emerged as the most abundant in all analyzed samples; its concentration in the edible part reached 1770.72 in Placoseps and 995.20 mg/kg in Darlise. Cooking increased rutin content in April-harvested asparagus to 1966.00 in Placoseps and 2042.44 mg/kg in Darlise, reflecting an increase of more than 2.5-fold compared to the respective values observed at the earlier harvest. Despite the substantial reduction in bioactive compounds observed during in vitro gastrointestinal digestion, a total of 146.95 to 454.58 mg/kg of bioaccessible compounds remaining available for potential intestinal absorption after digestion across both cultivars and harvest periods. These results provide a greater understanding of the behavior of polyphenol-rich vegetables and underscore the importance of simulating gastrointestinal processes when assessing the health-promoting potential of bioactive compounds. Full article

Pomegranate is recognized for its health benefits, primarily due to its polyphenols and metabolites, such as urolithins (Uro-A), produced via colonic fermentation of ellagic acid (EA). These compounds make pomegranate a functional food with the potential to modulate chronic disease risk factors and enhance gut health by modulating microbiota. The aims of this study were (1) to evaluate the effect of in vitro digestion and fermentation mimicking human digestive processes on the total phenolic content (TPC) and the antioxidant capacity of a standardized pomegranate extract (PE), (2) to assess the effect of the digested PE on the functionality of gut microbiota isolated from healthy and diseased subject fecal materials via short-chain fatty acid (SCFA) determination, and (3) to measure Uro-A production using UHPLC Q-Orbitrap HRMS. The in vitro digestion and fermentation processes resulted in a significant increase in the TPC, while the antioxidant capacity was considerably reduced. Following the in vitro digestion process, the TPC increased from 232 ± 16 to 1656 ± 34 g GAE/g of PE. Moreover, the TPC in the fermented samples was calculated as 6139 ± 458 g GAE/g for the microbiota of healthy adults and 8375 ± 1388 g GAE/g for the microbiota of healthy children, compared to 1657 ± 34 g GAE/g for the non-fermented samples. The PE exerted a modulatory effect on gut microbiota functionality, as reflected by an increasing concentration of SCFAs, especially lactic acid. Overall, these data suggest that pomegranate might contribute to gut health and could be a candidate for further studies in view of its possible use as a prebiotic ingredient. Further research, including clinical studies, is needed to confirm these findings and explore the potential application of pomegranate extract as a functional ingredient in nutraceuticals and functional foods aimed at improving gut health. Full article

Potato common scab, caused by Streptomyces species, is a widespread soil-borne disease that poses a significant threat to potato cultivation globally. In this study, a Bacillus velezensis D7-8 strain was isolated from a potato. This endophytic bacterium exhibited broad-spectrum antifungal activity, and pot trials demonstrated that the D7-8 strain effectively controlled potato common scab with an efficacy of 42.07%. The complete genome sequence of the D7-8 strain was sequenced and subsequently identified as B. velezensis through multiple bioinformatic methods, primarily through structural variation analysis of whole-genome sequences. The machine learning method predicted that the expression profiles of colinear genes among closely related Bacillus species were highly consistent. Metabolite analysis of crude extracts using ultra-high-performance liquid chromatography coupled with quadrupole-Orbitrap high-resolution mass spectrometry (UPLC-Q-Exactive HRMS) revealed that D7-8 produces bioactive compounds, including surfactin and fengycin, both known for their antimicrobial properties. This study elucidates the antagonistic effect of B. velezensis D7-8 against Streptomyces acidiscabies and provides a valuable reference for future research on accurate microbial identification. Full article

Background: Kukeya tablets (KYs), a traditional ethnic medicine prescription, are widely used to treat migraines and eye ailments in China. Despite their extensive clinical use, current knowledge on their therapeutic material basis is limited to a few major compounds, whereas certain minor ones have rarely been investigated. This study was conducted to screen and characterize the chemical components of KYs. Methods: A rapid and effective UHPLC-Q-Orbitrap-HRMS method was established. A mass spectrometry qualitative analysis strategy for KYs was developed, including in-house library matching, accurate molecular mass and elemental composition matching, and MS/MS fragmentation rule elucidation. Results: In total, 144 compounds were identified in KYs, including 36 anthrones and anthraquinones, 36 chromones, 25 triterpenes, 12 resin glycosides, 12 phenylpyrones, 10 phenolic acids, 4 flavonoids, 2 lignans, and 7 others. Meanwhile, the identified compounds were effectively classified into nine chemical classes. Among them, 11 compounds were identified for the first time and the identities of 22 compounds were accurately confirmed using reference substances. Conclusions: The results obtained benefit the understanding of the therapeutic basis of KYs, significantly promote the quality control of KYs, and elucidate potential effective components of other traditional medicines. Full article

Ultra-high-performance liquid chromatography coupled to Orbitrap high-resolution mass spectrometry (UHPLC-HRMS) was recently employed in many fields to obtain a rapid characterization of plant extracts. Kigelia africana (family Bignoniaceae) is a quintessential African herbal medicinal plant with immense indigenous medicinal and non-medicinal applications. The aim of the present research was to obtain an in-depth metabolite profiling of the K. africana stem bark extract using UHPLC-HRMS and to conduct a preliminary screening of its anticancer activity against a panel of malignant human cell lines of different origin. The UHPLC-HRMS analysis revealed 63 secondary metabolites including phenolic acids, gallo- and ellagitannins, iridoids, naphthoquinones, and anthraquinones. A total of 34 of all annotated compounds are reported for the first time in K. africana stem bark. The studied profile was dominated by trimethylellagic acid, dimethylellagic acid isomers, and ellagic acid. In all tumor models, we established a pronounced inhibition of cell growth in a mostly dose-dependent manner, with IC₅₀ values ranging near and well below (4–30 µg/mL) the lowest treatment concentration of 25 µg/mL. The established cytotoxicity profile of the K. africana extract, highly biased toward malignantly transformed but not normal cells, suggests specific modulation of defined molecular tumor targets. This study revealed K. africana stem bark as a new source of gallo- and ellagitannins, and highlighted the studied herbal drug as an antiproliferative agent with potential pharmaceutical application. Full article

Four aromatic and therapeutic plants, Thymus vulgaris, Rosmarinus officinalis, Pimpinella anisum, and Foeniculum vulgare, were examined in this comparative study. The objectives were to assess its phytochemical composition; polyphenol, flavonoid, and tannin content; antioxidant and antibacterial activity; bioactive molecule identification; and critical trace element quantification. Its aqueous and organic extracts were examined, focusing on ethanolic extracts. The ethanolic extract’s ability to neutralize free radicals was validated by phytochemical studies and antioxidant tests, underscoring their role in preventing oxidative stress. An Ultra-Performance Liquid Chromatography—High-Resolution Mass Spectrometry Orbitrap Exploris 120 (UPLC–HRMS Orbitrap) was used to identify the bioactive chemicals, and the results showed a variety of compounds having antibacterial and antioxidant properties. The important trace elements found in these plants were also measured using a Graphite Furnace-Atomic Absorption Spectrometer (GF-AAS). These components are essential to the biological characteristics of the plants, especially their antioxidant and antibacterial capacities. Among the aqueous extracts, it was observed that Rosmarinus officinalis and Foeniculum vulgare exhibited a MIC of 3.91 µg/mL against Staphylococcus. Additionally, R. officinalis also demonstrated a MIC of 3.91 µg/mL against Escherichia coli. All of the data were interpreted and shown using principal component analysis. The results were grouped and explained using this statistical method, which revealed a strong association between the abundance of antibacterial and antioxidant chemicals in the four plants under investigation. Full article

This study investigates the toxic effects of ketamine on juvenile zebrafish, driven by increasing concerns over ketamine’s prevalence and its potential neurotoxic effects that may disrupt behavior and metabolism. Employing a high-throughput behavior tracking system, the research analyzed the locomotor activity of 6-day post-fertilization (6 dpf) zebrafish exposed to various concentrations of ketamine. The integration of behavioral analysis with metabolic profiling was a notable innovation, as it establishes a comprehensive understanding of ketamine’s effects on both acute behavioral inhibition and metabolic responses. The findings reveal that ketamine exposure significantly inhibits locomotor activity in juvenile zebrafish, with these effects becoming more pronounced at higher concentrations. Additionally, the detection of normethketamine, the primary metabolite of ketamine, using UPLC-LTQ/Orbitrap HRMS, confirms the zebrafish’s ability to metabolize the drug. This underscores the utility of zebrafish as a model organism for studying the impact of ketamine on behavior and metabolism, providing valuable insights that may extend to other vertebrates. Full article

Food is essential for human survival; however, food can be an important route of exposure to contaminants. This study investigated the presence and distribution of anthropogenic contaminants in food consumed by families with small children in South Florida, United States, evaluating seasonal and socio-economic variabilities in chemical composition. QuEChERS protocols, followed by non-targeted analysis (NTA) using an LC-Orbitrap HRMS system, were used for the comprehensive screening of organic contaminants. The compounds were annotated and identified with the Compound Discoverer (CD) software, and contaminant distributions were analyzed using boxplots and Principal Component Analysis (PCA). The results showed significant seasonal and socio-economic differences in contaminant distributions (p < 0.05). In the wet season, a predominance of polymers and surfactants, such as dodecanedioic acid and N-dodecylacrylamide, were found in food, which might be due to increased transport of industrial pollutants during increased precipitation, while plasticizers (e.g., bis(2-ethylhexyl) phthalate) and drugs (e.g., warfarin) were more prevalent during the dry season, which could be related to less dilution effects in this period. A higher abundance of 1-nitrosopiperidine, present in cured meats, was noted in food from upper socio-economic classes, while the lower class showed higher abundance of benzocaine, a common topical anesthetic. Full article

Background/Objectives: Hemerocallis citrina Baroni (HCB) is a traditional herb for the treatment of depression in China. However, the active constituents and the underlying mechanisms of its antidepressant effects remain unclear. The aim of this study was to identify the bioactive constituents of HCB and elucidate its underlying mechanism for the treatment of depression. Methods: The constituents of HCB were systematically analyzed using UHPLC-Q-Orbitrap HRMS. Its antidepressant effect was evaluated by chronic unpredictable mild stress (CUMS)-induced depression. The mechanism of HCB in treating depression was investigated through network pharmacology and molecular docking. Subsequently, its potential mechanism for the treatment of depression was carried out by RNA sequencing. Finally, the mechanism was further verified by Western blot. Results: A total of 62 chemical constituents were identified from HCB using UHPLC-Q-Orbitrap HRMS, including 17 flavonoids, 11 anthraquinones, 11 alkaloids, 10 caffeoylquinic acid derivatives, five phenolic acids, five triterpenoids, and three phenylethanosides, 13 of which were identified as potential active constituents targeting 49 depression-associated proteins. Furthermore, HCB was found to significantly reduce cognitive impairment, anxiety-like behavior, and anhedonia-like behavior. The expression levels of 5-hydroxytryptamine (5-HT), dopamine (DA), and brain-derived neurotrophic factor (BDNF) were elevated in the hippocampal CA3 region. Results from network pharmacology and transcriptomics indicated that the PI3K/Akt/CREB signaling pathway is essential for the therapeutic effects of HCB on depression. Research in the field of molecular biology has conclusively demonstrated that HCB is associated with an increase in the expression levels of several important proteins. Specifically, there was a notable upregulation of phosphorylated PI3K (p-PI3K) relative to its unphosphorylated form PI3K, as well as an elevation in the ratio of phosphorylated Akt (p-Akt) to total Akt. Additionally, the study observed increased levels of phosphorylated CREB (p-CREB) compared to its unphosphorylated CREB. Conclusions: This study provides compelling evidence that HCB possesses the ability to mitigate the symptoms of depression through its influence on the PI3K/Akt/CREB signaling pathway. HCB could be developed as a promising therapeutic intervention for individuals struggling with depression, offering new avenues for treatment strategies that target this particular signaling mechanism. Full article

Although edible flowers have been historically principally used due to their visual appeal and smell, the world is discovering their value as innovative and natural sources of bioactive compounds. Cucurbita pepo L. (CpL), a plant from the Cucurbitaceae family, is widely cultivated for its edible fruits and flowers, which are rich in polyphenols and carotenoids—compounds known for their potent antioxidant and anti-inflammatory properties. Despite their potential, the use of CpL flowers for skin-related applications remains underexplored. This study aimed to comprehensively analyze CpL flower extract (CpLfe), focusing on its polyphenolic and carotenoid content using, for the first time, advanced UHPLC-Q-Orbitrap HRMS and HPLC-DAD analysis. CpLfe highlighted remarkable antioxidant activity according to the DPPH, ABTS, and FRAP tests. CpLfe showed significantly reduced intracellular ROS in HaCaT (23%, p < 0.05) and protected against UVB-induced damage by lowering MMP-1 expression. CpLfe also upregulated genes crucial for skin hydration (AQP3) and barrier function (CerS2, CerS4, and CerS6). A placebo-controlled, randomized clinical trial further validated CpLfe efficacy, demonstrating marked improvements in moisture retention, wrinkle reduction, and collagen production in women aged 35–55. These findings suggested that CpL flowers could be a source of bioactive compounds recovered from edible flowers able to improve the major skin aging and photoaging features. Full article

A new method for simultaneous determination by liquid chromatography coupled with high resolution mass spectrometry (UHPLC-HRMS/MS) of 14 paralytic shellfish poisoning toxins (PSP), that is, Saxitoxin, Neosaxitoxin, Gonyautoxins and their respective variants, in bivalve molluscs, is herein described. The samples were extracted by acetic acid solution, then analysed by UHPLC coupled with a Q-Exactive Orbitrap Plus high resolution mass spectrometer, by electrospray ionization mode (ESI) with no further clean up step. The analysis was carried out by monitoring both the exact mass of the molecular precursor ion of each compound (in mass scan mode, resolution at 70,000 FWHM) and its respective fragmentation patterns (two product ions) with mass accuracy greater than 5 ppm. The analytical performance of the method was evaluated calculating trueness, as mean recoveries of each biotoxin, between 77.8% and 111.9%, a within-laboratory reproducibility (RSD_R) between 3.6% and 12.2%, the specificity, the linearity of detector response, and the ruggedness for slight changes The results of the validation study demonstrate this method fits for the purposes of the official control of PSP toxins in molluscs. The results of two years of monitoring in local mussel farms are also reported, showing that no significant concerns for food safety in the monitored productions. Full article

Background/Objectives: Corema album berries are edible fruits from the Iberian Atlantic coast, characterized by a rich polyphenolic composition, which endows their juice with potential protective effects against neurodegeneration. This study aimed to evaluate the potential of the relatively lesser-known C. album berries as a novel neuroprotective agent against neurodegenerative diseases. Methods: The phenolic compounds of the juice were characterized using UHPLC-HRMS (Orbitrap). The SH-SY5Y neuroblastoma line was used to determine the preventive effect of the juice against H₂O₂-induced oxidative stress. Furthermore, neuronal cells were differentiated into dopaminergic and cholinergic lines and exposed to 6-hydroxydopamine and okadaic acid, respectively, to simulate in vitro models of Parkinson’s disease and Alzheimer’s disease. The ability of the juice to enhance neuronal viability under toxic conditions was examined. Additionally, its inhibitory effects on neuroprotective-related enzymes, including MAO-A and MAO-B, were assessed in vitro. Results: Phytochemical characterization reveals that 5-O-caffeoylquinic acid constitutes 80% of the total phenolic compounds. Higher concentrations of the juice effectively protected both differentiated and undifferentiated SH-SY5Y cells from H₂O₂-induced oxidative damage, reducing oxidative stress by approximately 20% and suggesting a dose-dependent mechanism. Moreover, the presence of the juice significantly enhanced the viability of dopaminergic and cholinergic cells exposed to neurotoxic agents. In vitro, the juice inhibited the activity of MAO-A (IC₅₀ = 87.21 µg/mL) and MAO-B (IC₅₀ = 56.50 µg/mL). Conclusions: While these findings highlight C. album berries as a promising neuroprotective agent, further research is required to elucidate its neuroprotective mechanisms in cell and animal models and, ultimately, in human trials. Full article

Over recent decades, the global occurrence of pesticide residues in aquatic environments has been a pivotal issue; however, their trace-level concentrations necessitate the establishment of ultra-sensitive and reliable analytical approaches. To this end, the present study describes the optimization and validation of an LC-HRMS-based method for the accurate determination of 18 pesticides in river and sea water, accompanied by a measurement uncertainty estimation. This method was applied to analyze 17 real samples from agriculture and aquaculture-impacted areas in Greece and Albania. Different solid-phase extraction (SPE) protocols were tested. For the analysis, cutting-edge Orbitrap MS technology and MS/MS fragmentation, along with the use of matrix-matched calibration curves, provided unprecedented accuracy (<5 ppm) and sensitivity for the confirmation of positive detections. Regarding method performance, exceptional linearity was obtained; the limits of quantification ranged from 1.7 ng L⁻¹ to 90 ng L⁻¹, recoveries varied from 61% to 96% in river water, while slightly higher recoveries (60–111%) were observed in seawater. In all cases, repeatability and intra-laboratory reproducibility were below 15%. The measurement expanded uncertainty (U′, k = 2) was estimated considering precision and bias. MU% values were lower than 50% in all cases, as recommended in SANTE guidelines and applied to the quantified results. The matrix effect study exhibited negative values (<20%) for all compounds. Application to real samples showed a low pesticide contamination load that should not be underestimated. Full article