Search Results (1,820)

Aralia chinensis L. has shown potential in breast cancer treatment, yet its pharmacodynamically active components and mechanisms remain undefined. To systematically identify the bioactive constituents absorbed into the bloodstream and elucidate their multi-target mechanisms against breast cancer, we employed ultra-high-performance liquid chromatography in conjunction with Q Exactive Orbitrap mass spectrometry (UHPLC-Q Exactive Orbitrap-MS) alongside serum pharmacochemistry to analyze the chemical constituents of total saponins derived from A. chinensis (TSAC) and to identify the blood-absorbed prototypes in a rat model. Network pharmacology predicted targets and pathways of serum prototypes, validated by molecular docking and in vitro experiments. We identified 38 triterpenoid saponins, 3 steroidal saponins, and 8 triterpenoids in TSAC, with 22 prototype compounds detected in serum. An integrative analysis encompassing 486 compound targets and 1747 genes associated with breast cancer elucidated critical pathways, notably the PI3K-Akt signaling pathway and resistance mechanisms to EGFR tyrosine kinase inhibitors. Molecular docking confirmed strong binding of araloside A and elatoside L to SRC, PIK3R1, PIK3CA, STAT3, and EGFR. In MCF-7 cells, TSAC suppressed proliferation and migration while downregulating Src, PI3K, and EGFR expression at the gene and protein levels. This study successfully identified TSAC’s serum-absorbed bioactive components and demonstrated their anti-breast cancer effects via multi-target mechanisms involving the Src/PI3K/EGFR axis, providing a crucial pharmacological foundation for developing A. chinensis-derived breast cancer therapies. Full article

Mycotoxin contamination in wheat, a staple food critical to human nutrition, poses significant public health concerns. This study investigated the natural occurrence of 17 mycotoxins in Tunisian durum wheat, assessed the influence of soil tillage practices on mycotoxin contamination, and performed an associated exposure risk assessment. A total of 167 wheat samples were randomly collected over two years (2021 and 2022) from fields managed under conventional tillage (CT) and no-tillage (NT) systems during both pre- and post-harvest periods. Mycotoxins were extracted using the QuEChERS method and quantified via UHPLC-MS/MS. The results demonstrated contamination by ZEN, DON, OTA, ENA1, ENB, and ENB1. Among regulated mycotoxins, OTA was the most prevalent, detected in 68 out of 167 samples with a mean concentration of 1.85 µg/kg. ZEN was the most abundant, detected in 65 samples with a mean concentration of 26.85 µg/kg, while DON was less frequently detected in 62 samples with a mean concentration of 0.68 µg/kg. Regarding emerging mycotoxins, ENB was the most prevalent and abundant, found in 51 samples with a mean concentration of 10.13 µg/kg; ENB1 and ENA1 were detected in 20 and 10 samples, with mean concentrations of 3.38 µg/kg and 1.69 µg/kg, respectively. Furthermore, mycotoxin concentrations varied according to agricultural practices. DON, ZEN, ENA1, ENB, and ENB1 showed higher frequencies and concentrations (ranging from 0.08 to 210.11 µg/kg) in samples collected during the 2021 pre-harvest period from NT fields. In contrast, OTA exhibited greater prevalence and higher concentrations (ranging from 2.33 to 9.78 µ/kg) in samples collected during the 2022 post-harvest period from CT fields. The Estimated Daily Intake (EDI) of mycotoxins by Tunisian adults was calculated based on contamination levels in raw durum wheat from fields under NT and CT practices, resulting in the following values (ng/kg bw/day), with the first value corresponding to NT samples and the second to CT samples: OTA (17.3; 20.8), ZEN (466.3; 194.0), DON (8.0; 7.56), ENA1 (4.30; 18.85), ENB (105.17; 121.08), and ENB1 (49.91; 40.91). Both the Margin of Exposure (MOE) values for OTA and the Hazard Quotients (HQ) for ZEN and DON exceeded established safety thresholds, indicating potential health risks for Tunisian adults. These findings highlight the urgent need to implement stricter mycotoxin regulations in Tunisia and enhance surveillance systems. Further research is warranted to elucidate the mechanisms by which soil tillage practices influence mycotoxin contamination and to develop targeted mitigation strategies to ensure food safety. Full article

Isothiocyanates (ITCs), which are derivatives of glucosinolates (GSLs) from Brassica plants, have been investigated as anticancer agents. An extensively studied anticancer ITC is sulforaphane, which is found in low amounts in Chinese cabbage. We aim to investigate the types and content of GSLs (precursors of ITCs with anticancer activity) in Chinese cabbage seeds. GSLs from Chinese cabbage seeds were isolated and purified using acidic Al₂O₃ column chromatography and preparative HPLC. GSL and ITC profiles were further identified using UHPLC-Q-TOF-MS. The antitumor activities of ITC (produced by exogenous enzymatic hydrolysis of GSLs) were evaluated in vitro. Seventeen GSLs and seven ITCs were identified, and the dominant GSLs were gluconapin, glucobrassicanapin, and progoitrin in Chinese cabbage seeds. High-purity gluconapin (>99%) was purified. The ITCs showed synergistic-, dose-, and time-dependent effects on the inhibition of HepG2 cells, and the key ITCs were 3-butenyl ITC, sulforaphane, and 2-phenylethyl ITC. The corresponding parent GSLs were gluconapin, glucoraphanin, and gluconasturtiin, respectively. 3-Butenyl ITC could significantly induce HepG2 cell proliferation (IC50 = 89.44 μg/mL) and apoptosis (p < 0.05). Our results suggested that Chinese cabbage seed could be a valuable source of natural antitumor ingredients. Full article

Objectives: A thorough understanding of pharmacokinetics and metabolism is critical during early drug development. This study investigates the absorption, distribution, metabolism, and excretion (ADME) profile of R14, a novel compound, using a combination of in vitro and in vivo approaches. Methods: In vitro studies included Caco-2 permeability assays, metabolic stability evaluations in liver microsomes and hepatocytes, and identification of CYP isoforms responsible for R14 metabolism. In vivo pharmacokinetic and metabolic profiling was conducted in rats following oral administration. R14 was quantified using UHPLC-MS/MS. Metabolites were identified using high-resolution UHPLC- QTOF MS/MS, and relative exposure was estimated using peak area-derived AUCs. Results: R14 exhibited low oral bioavailability (13.4%) and high systemic clearance (2.63 L/h/kg), indicating high hepatic extraction. A total of 21 plasma and 38 urine metabolites were identified. Major metabolic pathways included initial hydroxylation and hydrogenation, followed by sequential methylation and Phase II conjugations (glucuronidation and sulfation). Key metabolites (M3, M4, M22, M38) accounted for the majority of systemic exposure. Less than 1% of the unchanged drug was excreted in urine, confirming extensive metabolism. Notably, discrepancies between in vitro and in vivo metabolite profiles suggested rapid further transformation of initial metabolites in vivo, which were not fully captured in vitro. Conclusions: This study demonstrates an efficient and integrated strategy for early-phase ADME characterization. The combined use of in vitro assays and in vivo studies, guided by advanced analytical techniques, provides a robust framework for understanding drug metabolism. These findings can inform drug optimization and help minimize risks in later stages of development. Full article

Wild blackberry (Rubus ulmifolius Schott) is a culinary and medicinal plant traditionally used to treat various ailments, including hypertension. This study evaluated the vasorelaxant effects of five crude leaf extracts of R. ulmifolius (hexane, dichloromethane, ethyl acetate, methanol, and aqueous), as well as the hypotensive and antioxidant activities of its methanolic extract (MERu), and analyzed its phytochemical profile. Crude extracts, obtained using a Soxhlet apparatus, were tested in vitro on isolated rat aortic rings precontracted with phenylephrine. The hypotensive effect of MERu was examined in vivo in normotensive rats, and its antioxidant activity was assessed using the DPPH assay. Total phenolic and tannin contents were quantified by the Folin–Ciocalteu and hide powder methods, respectively, while UHPLC-MS was used to identify its phytochemicals. All crude extracts induced concentration-dependent vasorelaxation, with MERu showing the strongest effect (59.31% relaxation at 10⁻¹ g/L). Intravenous MERu induced significant blood pressure reductions in rats, starting at 1 mg/kg. At 20 mg/kg, systolic, diastolic, and mean arterial pressures dropped by 38.61%, 51.58%, and 45.19%, respectively. MERu also demonstrated potent antioxidant activity and was rich in polyphenols, particularly tannins. Sixteen compounds were identified, notably rubanthrone A, a galloyl-bis-HHDP glucose derivative, ellagic acid, and quercetin-3-O-β-D-glucuronide. These results suggest that R. ulmifolius may have therapeutic potential for hypertension and exhibits promising characteristics as a functional food. Full article

Background: Molybdenum (Mo) is an essential trace element for animals, but too much intake can cause adverse effects. Due to the metabolic characteristics of goats and other ruminants, they are more susceptible to the cumulative effects of Mo toxicity. A high Mo intake can cause multi-organ toxicity in ruminants, but the mechanism of damage to the pancreas is still unclear. The aim of this study was to systematically analyze the key regulatory pathways of pancreatic injury induced by Mo in goats using a metabolomics approach. Methods: Twenty male Yudong Black goats (22.34 ± 1.87 kg, six months) were randomly divided into a control group (fed a basal diet) and the Mo group (fed a basal diet supplemented with 50 mg·kg⁻¹ Na₂MoO₄·2H₂O). After 60 days of continuous feeding, their pancreatic tissues were collected and the mineral elements, antioxidant capacity, and inflammatory factors were examined. Untargeted metabolomics based on HILIC UHPLC-Q-EXACTIVE MS was used to analyze changes in metabolites. The core regulatory mechanisms were revealed by KEGG enrichment analysis. Results: The results demonstrated that goats in the Mo group showed obvious clinical signs, such as lethargy, loss of appetite, and unsteady gait. The pancreatic tissue of goats in the Mo group exhibited significantly elevated levels of Mo and copper, accompanied by a marked reduction in antioxidant capacity and concurrent increases in inflammatory cytokine levels. Between the Mo group and control group, 167 differentially expressed metabolites were identified. KEGG enrichment analysis showed that it disrupted multiple metabolic pathways, including glycine, serine, and threonine metabolism, cysteine and methionine metabolism, and butanoate metabolism. Conclusions: This study mainly revealed, at the metabolomics level, that Mo exposure would disrupt the metabolic pathways related to antioxidant capacity in goat pancreata. It provides new insights into the molecular mechanisms of Mo-induced pancreatic injury in goats. Full article

Origanum vulgare L. is an important aromatic plant traditionally used in folk medicine since ancient times. Its growing interest for the scientific community is mainly attributed to its distinctive chemical profile, which includes bioactive compounds, such as polyphenols (phenolic acids and flavonoids) and volatile compounds (essential oil). These components collectively contribute to oregano’s wide spectrum of biological activities. In this study, the volatile components of the essential oil (WEO_OR) and the polyphenolic fraction of the methanolic extract (ME_OR) obtained from leaves and inflorescences of wild Origanum vulgare collected in central Italy were characterized using GC-MS and UHPLC-DAD, respectively. Carvacrol was identified as the major compound in the essential oil, while rosmarinic acid was predominant in the methanolic extract. A comparative analysis was also carried out with a commercially available essential oil (CEO_OR), aiming to evaluate potential differences in chemical composition and antioxidant activity (DPPH, ABTS, and FRAP assays). ME_OR showed the strongest antioxidant activity (DPPH IC₅₀ = 0.052 mg mL⁻¹; ABTS = 3.94 mg TE mL⁻¹; FRAP = 30.58 mg TE g⁻¹), followed by CEO_OR (DPPH IC₅₀ = 0.45 mg mL⁻¹; ABTS = 9.57 mg TE mL⁻¹; FRAP = 7.33 mg TE g⁻¹), while WEO_OR displayed the lowest values (DPPH IC₅₀ = 1.54 mg mL⁻¹; ABTS = 0.10 mg TE mL⁻¹). Furthermore, ME_OR and WEO_OR were tested in vitro using the human hepatoblastoma cell line HepG2 to assess their potential biological activities related to cell survival and oxidative stress. The results indicated that at the tested doses, neither the ME nor the EO showed significant toxicity, as evidenced by the unchanged proliferation rate of HepG2 cells. However, the ME at low doses (50 and 100 μg mL⁻¹) and the EO (0.005%), administered as a pre-treatment, exhibited a protective effect against oxidative stress, as inferred from the reduction in 8-OHdG levels, a marker of oxidative damage to nucleic acids. Full article

This study aimed to evaluate the protective effects of Inula japonica leaf extract against PM₁₀-induced oxidative stress in normal human keratinocytes. Keratinocytes were pretreated with various concentrations of Inula japonica leaf extract and subsequently exposed to PM10. Cell viability, ROS production, gene and protein expression (qRT-PCR and Western blot), and UHPLC-MS profiling were assessed. Network pharmacology analysis was conducted using database-predicted compounds of Inulae Flos. The extract significantly reduced PM₁₀-induced ROS generation and restored the expression of epidermal barrier-related genes such as loricrin. It also inhibited phosphorylation of MAPKs (ERK, p38) and modulated apoptotic and inflammatory markers including Bax, p53, MMP-9, and COX-2. UHPLC-MS analysis identified eight compounds not previously reported in our earlier study, which may contribute to the extract’s protective effects. Inula japonica leaf extract exerts protective effects against PM₁₀-induced skin damage by reducing oxidative stress and inflammation in keratinocytes. These findings support its potential as a therapeutic candidate for pollution-related skin disorders. Full article

Veterinary drug residues in aquatic products are often overlooked, yet they pose significant environmental risks and potential threats to human health. In this study, a rapid and sensitive analytical method was developed for the simultaneous determination of nine commonly used macrolide antibiotics in largemouth bass (Micropterus salmoides) muscle using ultrahigh-performance liquid chromatography–tandem mass spectrometry (UHPLC-MS/MS). Sample extraction was performed using 80% acetonitrile in water, followed by purification with Cleanert MAS-Q cartridges. Chromatographic separation was achieved on a Waters ACQUITY UPLC BEH C₁₈ column (50 mm × 2.1 mm; 1.7 μm), equipped with a Waters VanGuard^TM BEH C₁₈ guard column (1.7 μm), using a mobile phase consisting of 0.1% formic acid in water and 0.1% formic acid in acetonitrile. Mass spectrometric detection was conducted in positive electrospray ionization mode (ESI⁺) using multiple reaction monitoring (MRM). The method demonstrated excellent linearity in the concentration range of 0.2–30 ng/mL, with determination coefficients (R²) exceeding 0.9980 for all analytes. Average recoveries ranged from 89.3% to 108.4%, with intraday and interday relative standard deviations (RSDs) of 2.9–11.6% and 4.1–12.5%, respectively. The limits of detection (LOD) and quantification (LOQ) for largemouth bass muscle were determined to be 0.4 μg/kg and 2.0 μg/kg, respectively. The decision limits (CC_α) and detection capabilities (CC_β) ranged from 2.13 to 215.71 μg/kg and 2.22 to 231.42 μg/kg, respectively. The developed method was successfully applied to the quantitative analysis of macrolide residues in 20 largemouth bass samples collected from local markets. Full article

Somatic hybridization represents a powerful tool for generating novel chemotypes with enhanced biosynthetic capabilities. This study provides the first comprehensive phytochemical characterization of interspecific somatic hybrids between Gentiana cruciata L. and Gentiana tibetica King ex Hook.f., two medicinally important yet regionally rare gentians. A total of 107 compounds were detected using UHPLC-DAD-ESI-MS³, of which 31 were identified as metabolites across eight phytochemical classes. Comparative profiling revealed that all hybrids retained a conserved core of iridoids and secoiridoids while integrating lineage-specific compounds and producing hybrid-specific metabolites not detected in either parent. Despite inheriting plastids from G. tibetica, hierarchical clustering showed that the phytochemical profiles of hybrid lines were more similar to G. cruciata, the donor of the nuclear genome. Quantitative analysis of the major secoiridoids, such as gentiopicroside, swertiamarin, and sweroside, demonstrated that several hybrid lines, particularly F30A-5 and F30A-6, matched or surpassed the biosynthetic output of G. tibetica, the more productive parent. These lines also exhibited elevated antioxidant capacity, underscoring their phytochemical and functional potential. Altogether, our findings show that somatic hybridization not only preserves but may amplify the secondary metabolite capacity of the parental genotypes, offering a viable platform for sustainable in vitro production of pharmacologically relevant secoiridoids. Full article

Pistachio nuts are among the 50 best foods with the highest antioxidant potential. They have a balanced content of mono- (~70%) and polyunsaturated (~20%) fatty acids, minerals, and bioactive compounds such as tocopherols, phytosterols, and phenolic compounds, which have shown rapid accessibility in the stomach. Pistachio consumption provides several health benefits, primarily due to its antioxidant properties and high content of essential nutrients. In this study, we analyzed the mineral composition, total phenolic content (TP), antioxidant activity (AA), and UHPLC/MS-MS polyphenolic profile of three Argentinian pistachio crops. Additionally, the physicochemical parameters and the elemental profiles of the growing soils were determined, as they influence mineral uptake and the synthesis of bioactive compounds in pistachio kernels. The TP was not significantly modified by the growing soils, with Crop3 presenting the highest TP content (276 ± 14 mg GA/100 g DW). Crop3 exhibited 18% higher TP content compared to Crop2. Similarly, FRAP values ranged from 28.0 to 36.5 mmol TE/100 g DW, with Crop1 showing a 30% increase compared to Crop2. DPPH values varied from 19.0 to 24.3 mmol TE/100 g DW, with Crop1 displaying 28% higher activity than Crop2. However, the polyphenolic profile was similar for all crops analyzed. Thirty compounds were identified; only Crop 1 contained the flavanone eriodyctiol and the isoflavone genistein, while the flavanone naringenin and the flavone luteolin were identified in Crop1 and Crop3. Regarding mineral content, the pistachio kernels mainly contained K, Ca, and Mg. Multivariate analyses revealed distinct elemental and antioxidant profiles among crops. LDA achieved classification accuracies of 77.7% for soils and 74.4% for kernels, with Pb, Zn, Cu, Rb, Sr, and Mn as key discriminants. CCA confirmed strong soil–kernel mineral correlations (r = 1), while GPA showed higher congruence between antioxidant traits and kernel composition than with soil geochemistry. These findings underscore the importance of soil composition in determining the nutritional quality of pistachio kernels, thereby supporting the beneficial health effects associated with pistachio consumption. Full article

The artichoke (Cynara cardunculus L. subsp. scolymus) is an endemic perennial plant of the Mediterranean area commonly consumed as food. It is known since ancient times for its beneficial properties for human health, among which its antioxidant activity due to polyphenolics stands out. In the frame of our ongoing studies aiming to highlight the biodiversity and the chemodiversity of natural resources, we investigated the phenolic and saponin content of the cultivar “Carciofo di Procida” collected at Procida, an island of the Gulf of Naples (Italy). Along with the edible part of the immature flower, we included in our analyses the stem and the external bracts, generally discarded for food consuming or industrial preparations. The LCMS quali-quantitative profiling of polyphenols (including anthocyanins) and cynarasaponins of this cultivar is reported for the first time. In addition to antioxidant properties, we observed a significant cytotoxic activity due to extracts from external bracts against human neuroblastoma SH-SY5Y cell lines with 43% of cell viability, after 24 h from the treatment (50 μg/mL), and less potent but appreciable effects also against human colorectal adenocarcinoma CaCo-2 cells. This suggests that the different metabolite composition may be responsible for the bioactivity of extracts obtained from specific parts of artichoke and foresees a possible exploitation of the discarded material as a source of beneficial compounds. Full article

One of the most important features of Metschnikowia pulcherrima is its strong antimicrobial activity against phytopathogens, which makes it a suitable candidate for use in biocontrol during crop cultivation. However, the mechanisms of its antimicrobial activity are not currently well understood. In this study, we used metabolomic methods to investigate the possible mechanisms of antimicrobial activity by M. pulcherrima against phytopathogenic fungi. First, we tested the antimicrobial activity of five selected isolates against eleven phytopathogenic molds. Based on the results, selected yeast–pathogen co-cultures were cultivated on liquid and solid media. The supernatants from the liquid co-cultures were analyzed using the UHPLC-QToF-UHRMS and MS/MS methods. Co-culture growth on solid agar media was examined using the LARAPPI/CI MSI method. The yeast exhibited strong antagonism toward the mold phytopathogens. The LARAPPI/CI MSI method revealed the presence of various compounds with potential antifungal activity. The complex UHPLC-QToF-UHRMS analysis confirmed that the metabolic response of M. pulcherrima depends on specific yeast–pathogen interactions. Full article

High semen quality is vital for reproductive success in the swine industry; however, seasonal fluctuations often compromise this quality. The molecular mechanism underlying these seasonal effects on semen quality remains largely unclear. This study employed untargeted metabolomic profiling of boar seminal plasma (SP) to identify metabolites and metabolic pathways associated with semen quality during the summer and winter months. Semen samples were collected from mature Duroc boars at a commercial boar stud and classified as Passed or Failed based on motility and morphology. SP from five samples per group was analyzed using ultra-high-performance liquid chromatography–mass spectrometry (UHPLC-MS). In total, 373 metabolites were detected in positive ion mode and 478 in negative ion mode. Several differentially expressed metabolites (DEMs) were identified, including ergothioneine, indole-3-methyl acetate, and avocadyne in the summer, as well as LysoPC, dopamine, and betaine in the winter. These metabolites are associated with key sperm functions, including energy metabolism, antioxidant defense, and capacitation. KEGG pathway analysis indicated enrichment in starch and sucrose metabolism, pyrimidine metabolism, and amino acid metabolism across the seasons. Overall, the results reveal that SP metabolomic profiles vary with the season, thereby influencing semen quality. The identified metabolites may serve as potential biomarkers for assessing semen quality and enhancing reproductive efficiency in swine production. Full article

Background/Objectives: Blunt cardiac injury (BCI) is a severe medical condition that may arise as a result of various traumas, including motor vehicle accidents and falls. The main objective of this study was to explore the role and underlying mechanisms of the TRPV4 gene in BCI. Elucidating the function of TRPV4 in BCI may reveal potential novel therapeutic targets for the treatment of this condition. Methods: Rats in each group, including the SD control group (SDCON), the SD blunt-trauma group (SDBT), the TRPV4 gene-knockout control group (KOCON), and the TRPV4 gene-knockout blunt-trauma group (KOBT), were all freely dropped from a fixed height with a weight of 200 g and struck in the left chest with a certain energy, causing BCI. After the experiment, the levels of serum IL-6 and IL-1β were detected to evaluate the inflammatory response. The myocardial tissue structure was observed by HE staining. In addition, cardiac transcriptome analysis was conducted to identify differentially expressed genes, and metabolomics studies were carried out using UHPLC-Q-TOF/MS technology to analyze metabolites. The results of transcriptomics and metabolomics were verified by qRT-PCR and Western blot analysis. Results: Compared with the SDCON group, the levels of serum IL-6 and IL-1β in the SDBT group were significantly increased (p < 0.001), while the levels of serum IL-6 and IL-1β in the KOBT group were significantly decreased (p < 0.001), indicating that the deletion of the TRPV4 gene alleviated the inflammation induced by BCI. HE staining showed that myocardial tissue injury was severe in the SDBT group, while myocardial tissue structure abnormalities were mild in the KOBT group. Transcriptome analysis revealed that there were 1045 upregulated genes and 643 downregulated genes in the KOBT group. These genes were enriched in pathways related to inflammation, apoptosis, and tissue repair, such as p53, apoptosis, AMPK, PPAR, and other signaling pathways. Metabolomics studies have found that TRPV4 regulates nucleotide metabolism, amino-acid metabolism, biotin metabolism, arginine and proline metabolism, pentose phosphate pathway, fructose and mannose metabolism, etc., in myocardial tissue. The combined analysis of metabolic and transcriptional data reveals that tryptophan metabolism and the protein digestion and absorption pathway may be the key mechanisms. The qRT-PCR results corroborated the expression of key genes identified in the transcriptome sequencing, while Western blot analysis validated the protein expression levels of pivotal regulators within the p53 and AMPK signaling pathways. Conclusions: Overall, the deletion of the TRPV4 gene effectively alleviates cardiac injury by reducing inflammation and tissue damage. These findings suggest that TRPV4 may become a new therapeutic target for BCI, providing new insights for future therapeutic strategies. Full article