Search Results (244)

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

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

Apples are one of the most popular fruits in Germany, valued for their regional availability and health benefits. When deciding which apple to buy, several characteristics are important to consumers, including the taxonomic variety, organic cultivation and regional production. To verify that these characteristics are correctly declared, powerful analytical methods are required. In this study, ultra-high performance liquid chromatography quadrupole time-of-flight mass spectrometry (UHPLC-Q-ToF-MS) is applied in combination with random forest to 193 apple samples for the analysis of various authentication issues. Accuracies of 93.3, 85.5, 85.6 and 90% were achieved for distinguishing between German and non-German, North and South German, organic and conventional apples and for six different taxonomic varieties. Since the classification models largely use different parts of the data, which is shown by variable selection, this method is very well suited to answer different authentication issues with one analytical approach. Full article

The widespread discharge of industrial and urban waste has led to significant increases in the environmental concentrations of numerous chemical substances. This work presents the development of a simple and environmentally friendly dispersive liquid–liquid microextraction (DLLME) method based on a hydrophobic natural deep eutectic solvent (NADES) for the determination of selected compounds from benzotriazole, benzothiazole, paraben, and UV filter families in wastewater samples. Of the twelve NADES formulations evaluated, those composed of a 4:1 molar ratio of thymol and menthol presented the highest extraction efficiencies. The influence of key experimental variables such as the pH of the aqueous sample, the ratio of NADES phase to sample volume, and the extraction time on the extraction efficiency was investigated using a multivariate optimization. Under optimal conditions, relative standard deviations below 15% and recoveries for spiked wastewater samples ranged between 82 and 108%, demonstrating the suitability of the method for routine water-quality monitoring. The sustainability and practicality of the developed method was evaluated using the assessment tools ChlorTox, AGREEprep, AGRRE, and BAGI, obtaining scores of 0.005 g in the NADES-DLLME method, 0.70, 0.52, and 72.5, respectively, demonstrating that the method is green and reliable. Full article

Background/Objectives: Vitamin D (VD) is metabolized in the body and plays a crucial role in regulating the antioxidant system. While exposure to heavy metals (HMs) inhibits VD activity, HMs can also be absorbed following VD stimulation. Despite differing views on the interaction between HM and VD activity, the effects of HM exposure on VD-related pathways have not been examined using metabolomics. This study aimed to investigate the impact of HM exposure on VD-related antioxidant activity under VD deficiency conditions using untargeted metabolic profiling. Methods: In this retrospective cohort study, 46 plasma samples were analyzed using ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UHPLC-QTOF/MS). Metabolic profiling was performed on two groups: individuals with severe VD deficiency and low HM exposure (SVDD–LHM) and those with VD deficiency and high HM exposure (VDD–HHM). Results: As a compensatory response to oxidative stress induced by HMs, VD-related antioxidant pathways may be associated with elevated levels of antioxidants, including bilirubin, eicosapentaenoic acid (EPA), and docosahexaenoic acid (DHA). In-creases in EPA and DHA were also linked to alterations in lipid metabolism, including diacylglycerol and phosphatidylcholine levels. DHA showed an area under the curve (AUC) of 0.850 (95% CI: 0.651–0.990), suggesting that DHA could serve as a potential biomarker for VD activity in response to HM exposure. Conclusions: The identified metabolites and metabolic pathways suggest that HM exposure may stimulate VD-related antioxidant activity, even under VD-deficient conditions. Full article

Throughout human history, wild plant resources have played an invaluable role, serving as critical sources of food, medicine, and industrial materials. This study examined the callus cultures of Cistanche deserticola Y.C. Ma, a medicinal desert plant, by subjecting them to abiotic stress under controlled in vitro conditions. The secondary metabolite profiles were then analyzed using GC-MS and qTOF-UHPLC-MS. The GC-MS analysis revealed several bioactive compounds of pharmaceutical interest, such as γ-sitosterol and homovanillyl alcohol. PhGs, including echinacoside and salidroside, were quantified for the first time across 16 callus samples exposed to various stress treatments. The application of 0.1% Na₂CO₃ for 50 days resulted in the highest accumulation of echinacoside (13,378.9 µg/mL), and heavy metal stress notably increased salidroside levels to 27.0 µg/mL. There was a clear correlation between callus pigmentation and metabolic activity: orange and white calli produced significantly more PhGs than dark calli. These results suggest that C. deserticola callus cultures could be a sustainable, controllable platform for producing high-value secondary metabolites. This reinforces the importance of wild plant resources in modern science and industry. Full article

A novel biodegradable food packaging material based on cassava thermoplastic starch (TPS) and polybutylene adipate terephthalate (PBAT) blends containing food preservatives was successfully developed using blown-film extrusion. This active packaging is designed to enhance the appearance, taste, and color of food products, while delaying quality deterioration. However, the incorporation of food preservatives directly influences consumer perception, as well as health and safety concerns. Therefore, this research aims to assess the risks associated with both intentionally added substances (IAS) and non-intentionally added substances (NIAS) present in the developed active packaging. The migration of both intentionally and non-intentionally added substances (IAS and NIAS) was evaluated using gas chromatography–mass spectrometry (GC-MS) and ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF-MS). Fifteen different volatile compounds were detected, with the primary compound identified as 1,6-dioxacyclododecane-7,12-dione, originating from the PBAT component. This compound, along with others, resulted from the polymerization of adipic acid, terephthalic acid, and butanediol, forming linear and cyclic PBAT oligomers. Migration experiments were conducted using three food simulants—95% ethanol, 10% ethanol, and 3% acetic acid—over a period of 10 days at 60 °C. No migration above the detection limits of the analytical methods was observed for 3% acetic acid and 10% ethanol. However, migration studies with 95% ethanol revealed the presence of new compounds formed through interactions between the simulant and PBAT monomers or oligomers, indicating the packaging’s sensitivity to high-polarity food simulants. Nevertheless, the levels of these migrated compounds remained below the regulatory migration limits. Full article

Background: The relationship between medicinal efficacy and the geographical environment in Thesium chinense Turcz. (T. chinense Turcz.), a traditional Chinese herb, remains systematically unexplored. This study integrates metabolomics, machine learning, and ecological factor analysis to elucidate the geographical variation patterns and regulatory mechanisms of secondary metabolites in T. chinense Turcz. from Anhui, Henan, and Shanxi Provinces. Methods: Metabolomic profiling was conducted on T. chinense Turcz. samples collected from three geographical origins across Anhui, Henan, and Shanxi Provinces. Machine learning algorithms (Random Forest, LASSO regression) identified region-specific biomarkers through intersection analysis. Metabolic pathway enrichment employed MetaboAnalyst 5.0 with target prediction. Antioxidant activity (DPPH/hydroxyl radical scavenging) was quantified spectrophotometrically. Environmental correlation analysis incorporated 19 WorldClim variables using redundancy analysis, Mantel tests, and Pearson correlations. Results: We identified 43 geographical marker compounds (primarily flavonoids and alkaloids). Random forest and LASSO regression algorithms determined core markers for each production area: Anhui (4 markers), Henan (6 markers), and Shanxi (3 markers). Metabolic pathway enrichment analysis revealed these markers exert pharmacological effects through neuroactive ligand–receptor interaction and PI3K-Akt signaling pathways. Redundancy analysis demonstrated Anhui samples exhibited significantly higher antioxidant activity (DPPH and hydroxyl radical scavenging rates) than other regions, strongly correlating with stable low-temperature environments (annual mean temperature) and precipitation patterns. Conclusions: This study established the first geo-specific molecular marker system for T. chinense Turcz., demonstrating that the geographical environment critically influences metabolic profiles and bioactivity. Findings provide a scientific basis for quality control standards of geo-authentic herbs and offer insights into plant–environment interactions for sustainable cultivation practices. Full article

Bee-collected pollen (BCP) can serve as an excellent enhancer of functional food bioactivity, particularly when it is fermented. The aim of this study was to prepare a novel kombucha-based beverage (KPE) enriched with fermented monofloral rapeseed (Brassica napus L.) BCP. To characterize the obtained samples, a proximate phytochemical composition analysis (including total phenolic and flavonoid content) was performed, as well as a detailed untargeted UHPLC-Q-ToF-MS profiling of phenolics and phenylamides. To biologically characterize KPE, antioxidant and antimicrobial activities were monitored. The total phenolic and flavonoid content, enhanced by the addition of BCP to the kombucha green tea beverage, was dose-dependent. The control sample showed a strong predominance of flavan-3-ols, distinguishing it from the KPE samples, where flavonol predominance and an increased content of phenolic acids were observed. Notably, the most significant markers of BCP were phenylamides, which were completely absent in the control. Although antioxidant activity was proximately highest in the control sample, KPE samples exhibited significantly improved antimicrobial activity. Full article

This study explores the microbiota and metabolite profile of raw cow’s milk, analyzing the correlation between them and their potential impact on consumer health, focusing on factors like cow’s feeding regimen (conventional vs. grazing), the herd’s geographical location (coastal vs. inland), and the thermal stability of raw milk samples. To achieve this, raw milk quality was assessed through mesophilic bacterial count and thermal stability using the 70° and 80° ethanol test. Metataxonomic analysis was performed using Illumina 16S rRNA hypervariable region sequencing, and untargeted metabolomics was conducted using UHPLC-Q/TOF MS/MS, followed by multivariate correlation analysis. All samples met quality standards. A total of 1258 prokaryotic ASVs were identified, spanning 48 bacterial phyla and one archaeal phylum, with 909 genera and 349 unidentified ASVs. Additionally, 70 metabolites were identified, including essential amino acids, vitamins, and bioactive compounds with antimicrobial, anti-inflammatory, and prebiotic properties. Significant correlations between bacterial diversity and metabolite profiles were observed. Feeding regimen and geographical location influence microbial composition and metabolite abundance, while thermal stability was linked to specific metabolites like oleamide and pyridoxal. These findings suggest that microbiota-derived metabolites can enhance the nutritional and functional value of milk. Further research is needed to understand how environmental and processing factors influence these dynamics. Full article

In this study, the reactivity of procyanidins and anthocyanins in young and aged Prokupac wines toward salivary proteins is investigated via SDS-PAGE and UHPLC-QTOF-MS to determine the differences between the phenolic compounds of red wine in relation to the aging process of wine. SDS-PAGE analysis revealed that procyanidins, flavanol-anthocyanin polymers, and ellagitannins in aged wine have strong affinities for salivary proteins, leading to the formation of insoluble complexes. By contrast, young wine contained predominantly procyanidins with high salivary protein affinity, as well as monomeric flavan-3-ols and anthocyanins, which mainly form soluble aggregates, while polymeric phenolics were less represented. Electrophoretic patterns further showed that seed-derived procyanidins mainly formed insoluble complexes with salivary proteins, whereas skin-derived anthocyanins tended to form soluble ones. The total content of all phenolic compounds quantified by UHPLC-QTOF-MS was 2.5 times higher in young wine than in aged wine, primarily due to the significantly greater abundance of malvidine-3-O-glucoside in young wine (eightfold higher level in young wine). Targeted UHPLC-QTOF-MS analysis of selected phenolics confirmed the electrophoretic results and showed a higher binding affinity of procyanidins in aged wine compared to young wine, as well as a higher percentage of procyanidin binding compared to anthocyanins, independent of the age of the wine. Sensory evaluation showed that aged wine had higher tannin quality scores, whereas young wine exhibited greater acidity and astringency, with bitterness being comparable between them. These results highlight the influence of wine aging on the interaction between phenolic compounds and salivary proteins and emphasize the dominant role of procyanidins in protein binding and the potential synergistic contribution of anthocyanins to mouthfeel perception. Full article

Background/Objectives: Alzheimer’s disease (AD) is the most common type of dementia, characterized by complex processes such as neuro-inflammation, oxidative damage, synaptic loss, and neuronal death. Carotenoids are among the potential therapeutic molecules that have attracted attention due to their neuroprotective properties, but their efficacy is limited mainly by their capacity to cross the blood–brain barrier (BBB). Results: The results showed that T. chuii extracts could protect neuronal cells from neurotoxic damage, especially against L-glutamate and H₂O₂. Moreover, the BBB permeability and the intestinal transport analyses revealed that fucoxanthinol, crocoxanthin, diatoxanthin, neoxanthin, violaxanthin, and prasinoxanthin have diverse permeabilities depending on the incubation time and the cell model used. Fucoxanthinol was the carotenoid with the highest and similar permeability in HBMEC cells (4.41%, 5.13%, and 18.94% at 2, 4, and 24 h, respectively) and Caco-2 cells (7.01%, 8.63%, and 18.36% at the same times), while crocoxanthin, diatoxanthin, and neoxanthin showed different kinetics. Methods: The neuroprotective potential of two extracts obtained from Tetraselmis chuii microalga were evaluated against Aβ1-42-, L-glutamate-, and H₂O₂-induced toxicities in SH-SY5Y cells. In addition, the BBB permeability and the intestinal transepithelial transport of the main carotenoids present in the extracts were evaluated and compared using two cell culture models, HBMEC and Caco-2 cells. For that aim, the transport of the bioactive molecules across the barriers was evaluated using UHPLC-q-TOF-MS after 2, 4, and 24 h of incubation. Conclusions: These findings indicate that T. chuii is a promising natural source of bioactive compounds to develop functional foods against neurodegenerative diseases. Full article

American ginseng (Panax quinquefolius L.) is a rare and valuable plant utilized for medicinal and culinary purposes, with its geographic origin and cultivation significantly affecting its quality and efficacy. However, the metabolic differences between cultivated and wild American ginseng are not well understood. An accurate and reliable method for tracing the origin and evaluating the quality of American ginseng is therefore urgently required. This study introduces a UHPLC-Q/TOF-MS-based comparative metabolomics and machine learning strategy for the rapid identification of wild and cultivated American ginseng. Both principal component analysis and hierarchical cluster analysis revealed distinct metabolic phenotypes between wild and cultivated American ginseng. Furthermore, the integration of univariate and multivariate statistical analyses identified eight differential metabolites in the ESI⁺ mode and three in the ESI^- mode, including seven ginsenosides. A potential ginsenosides marker panel was used to construct five machine learning models to assist in diagnosing the metabolic phenotypes of American ginseng. The Random Forest model, based on the eight differential metabolites in the ESI⁺ mode, achieved a 100% classification rate in both test and validation sets for distinguishing between wild and cultivated American ginseng. This study highlights the feasibility and application of our artificial intelligence-driven comparative metabolomics strategy for cultivar identification and geographic tracing of American ginseng, offering new insights into the molecular basis of metabolic variation in cultivated American ginseng. Full article

Atherosclerosis is a persistent inflammatory disorder influenced by oxidative stress and lipid imbalances, and it continues to be a major contributor to cardiovascular diseases. Rich in catechins and flavonoids, green tea pressurized hot water extract (GPHWE) demonstrated potent antioxidant activity through DPPH, ABTS, hydroxyl, and nitric oxide scavenging assays. In vitro, GPHWE protected RAW264.7 macrophages from oxidized LDL (Ox-LDL)-induced cytotoxicity and apoptosis by mitigating oxidative stress and enhancing cell survival. Animal studies using mice fed a high-fat diet (HFD) revealed notable improvements in lipid profiles, including decreases in total cholesterol, LDL, the atherosclerosis index (AI), the coronary risk index (CRI), and triglycerides, as well as lower levels of malondialdehyde (MDA), an indicator of oxidative stress. These results were comparable to those achieved with Simvastatin. Molecular docking studies indicated strong binding affinities of catechins to essential targets such as LOX-1, HMG-CoA reductase, caspase-3, and Nrf2, implying that the mechanisms of GPHWE involve antioxidant properties, regulation of lipids, and stabilization of plaques. The catechins of GPHWE, including epigallocatechin gallate (EGCG), epicatechin gallate (ECG), and epigallocatechin (EGC), were tentatively identified through qualitative analysis performed by UHPLC-QTOF-MS. This comprehensive approach positions GPHWE as a promising natural remedy for preventing atherosclerosis and reducing cardiovascular risk. Full article

Colostrum is a nutrient-dense food rich in proteins, immune modulators, and growth factors essential for neonatal development. Its lipids serve as a key energy source and facilitate cellular functions. While yak colostrum (YC) and buffalo colostrum (BC) exhibit high nutritional value, their lipid compositions remain unclear. This study employed lipidomics to analyze and compare the lipid profiles of YC and BC with cow colostrum (CC), identifying 872 lipids across 33 subclasses. Differential analysis revealed 137, 100, and 131 lipids, with significant expression differences between YC vs. CC, BC vs. CC, and YC vs. BC, respectively. YC exhibited higher levels of α-linolenic acid and docosahexaenoic acid, suggesting a potential role in enhancing neurodevelopment and cognitive function. In contrast, the upregulation of specific lipid components in BC, such as phosphatidylethanolamine (PE) and phosphatidylinositol (PI), coupled with the downregulation of specific lysophosphatidylcholine (LPC) and lysophosphatidylethanolamine (LPE), indicated a potential benefit for lipid metabolism and inflammatory regulation. These findings suggest that YC may be particularly suitable for neonates requiring enhanced energy support, while BC may offer advantages in lipid metabolism modulation. The study provides critical insights into the distinct lipid compositions of YC and BC, laying a scientific foundation for the development of tailored nutritional supplements. These results also hold significant implications for the dairy industry, driving innovation and optimization of colostrum-based products to meet diverse nutritional demands. Full article