Search Results (160)

Background: Chronic nonbacterial prostatitis (CNP), the major subset of chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS), imposes a substantial global burden yet lacks satisfactory therapies. Maizibizi Wan (MZBZ) has long been used clinically for prostatitis, but its pharmacodynamic substance basis and mechanisms remain unclear. Methods: Ultra-high-performance liquid chromatography–Q-Orbitrap high-resolution mass spectrometry (UHPLC-Q-Orbitrap-HRMS) coupled with Global Natural Products Social Molecular Networking (GNPS) molecular networking profiled MZBZ constituents and rat plasma–exposed prototype components and metabolites was used. Based on blood-absorbable components, network pharmacology predicted core targets/pathways; representative interactions were validated by molecular docking. A λ-carrageenan–induced CNBP rat model underwent histopathology (H&E), serum cytokine assays (TNF-α, IL-1β, IL-6/IL-17), immunohistochemistry (COX-2, TNF-α, MMP-9), and Western blotting (P-p65/p65, p-AKT/AKT, COX-2, TGF-β1, BCL2). Results: A total of 188 chemical constituents were identified in MZBZ (79 flavonoids, 38 organic acids, 30 alkaloids, 15 phenylpropanoids, 7 steroids, 4 phenylethanoid glycosides, 15 others). A total of 35 blood-absorbable components (18 prototype components, 17 metabolites) were identified, mainly involving Phase I oxidation and Phase II glucuronidation/sulfation. Network analysis yielded 54 core targets enriched in NF-κB and PI3K/AKT signaling and apoptosis. Docking indicated stable binding of key flavonoids to COX-2, NFKB1, TNF, IL-6, and BCL2. In vivo, MZBZ ameliorated prostatic inflammation, reduced serum TNF-α/IL-1β/IL-6/IL-17 (p < 0.05 or p < 0.01); decreased P-p65/p65, p-AKT/AKT, COX-2, and TGF-β1; and increased BCL2 in prostate tissue. Conclusions: MZBZ exerts anti-CNBP effects via multi-component synergy (prototypes + metabolites) that suppresses inflammatory cytokines, modulates apoptosis, and inhibits NF-κB and PI3K/AKT pathways. These findings provide a mechanistic basis and quality control cues for the rational clinical use of MZBZ. Full article

Fermentation, one of the oldest food processing techniques, is known to play a pivotal role in improving the nutritional and functional characteristics of cereals, with positive implications for gut health and overall well-being. The present study aims to examine the phenolic acids, peptides, and potential bioactive properties of 2 novel sorghum-based fermented beverages, Niselo and Delishe. A total of 48 phenolic compounds were identified through targeted and untargeted Ultra-High Performance Liquid Chromatography coupled with a Quadrupole Orbitrap High-Resolution Mass Spectrometer (UHPLC–Q-Orbitrap HRMS) analyses, revealing a higher content of phenolic acids in Niselo and a prevalence of flavonoids in Delishe. Niselo exhibited enhanced in vitro cytoprotective and reactive oxygen species (ROS)-scavenging activity and displayed a clear cytoprotective effect against hydrogen peroxide-induced oxidative stress in Caco-2 cells. Proteomic profiling using tryptic digestion revealed that Niselo has a substantial abundance of fragments of peptides matching several stress-related and antioxidant proteins, indicating a superior stress-response and/or defense capability. Overall, these findings highlight the functional potential of sorghum-based fermented beverages, supporting their role as health-promoting products. Full article

Background/Objectives: The biogenic synthesis of silver nanoparticles (AgNPs) is a promising alternative method, driven by the presence of metabolites in plant matrices capable of acting as reducing and stabilizing agents. Seasonality is a key factor that influences the phytochemical composition of plants and can directly impact the yield, physicochemical characteristics, stability, and bioactivities of the obtained AgNPs. This study aimed to synthesize AgNPs using aqueous extracts from Paullinia cupana leaves collected during dry and rainy seasons, prepared by two different methods (agitation or infusion), to evaluate the impact of these variables on the biosynthesis and properties of the nanostructures. Methods: The extracts were characterized by UHPLC-HRMS/MS, and their total phenolic compound (TPC) content and antioxidant potential against DPPH and ABTS radicals were determined. The AgNPs were characterized by UV/Vis spectrophotometry, dynamic light scattering (DLS), zeta potential (ZP), nano-particle tracking analysis (NTA), transmission electron microscopy (TEM), and energy-dispersive X-ray spectroscopy (EDX). Results: The metabolic profile results showed a predominance of alkaloids and flavonoids in all extracts, with greater phytochemical diversity in samples prepared by infusion. TPC indicated superior phenolic extraction in extracts prepared by infusion during the rainy season, correlating with greater antioxidant potential via the elimination of free radicals. The evolution of AgNP synthesis was accompanied by a gradual change in the color of the suspensions and the formation of plasmon bands between 410 and 430 nm, characteristic of spherical AgNPs. The nanostructures presented hydrodynamic diameters between 37.49 and 145.5 nm, PdI between 0.222 and 0.755, and Zeta potential between −11.3 and −39.9 mV, suggesting satisfactory colloidal stability. Morphological analyses revealed predominantly spherical particles with average diameters ranging from 33.61 to 48.86 nm and uniform distribution, while EDX spectra confirmed the presence of silver. Conclusions: Thus, our results demonstrate that both seasonality and the method of extract preparation influence the phytochemical composition and, consequently, the morphology, stability, and optical properties of AgNPs, with subtle emphasis on collections made during the rainy season and extracts prepared by infusion. Such knowledge contributes to the advancement of more reproducible and purpose-oriented syntheses in the field of green nanotechnology, enabling applications in various sectors. Full article

Goji berries (Lycium barbarum L.) are valued for their nutritional and medicinal properties; however, the systematic biochemical impact of drying on their quality remains poorly understood. This study applied an untargeted metabolomics approach based on UHPLC-HRMS and AntDAS to profile metabolic changes during sun-drying. Multivariate analyses (PCA and PLS-DA) revealed distinct time-dependent clustering, indicating significant shifts in the metabolome. Key metabolites, including betaine, galactose, and trans-ferulic acid, increased significantly (p < 0.05), whereas choline, allantoin, and huperzine isomers decreased. Pathway analysis highlighted glycine, serine, threonine, galactose, and phenylpropanoid metabolism as the central pathways that were affected. These differential metabolites could potentially be used as quality biomarkers. Our findings establish untargeted metabolomics as an effective tool for elucidating the evolution of goji berry quality during drying, offering a theoretical basis for process optimization. Full article

Background/Objectives: Land management practices strongly influence soil biochemical processes, yet conventional soil measurements often overlook dynamic small-molecule variation underlying nutrient cycling and microbial activity. This study aimed to evaluate whether MS¹-based untargeted metabolomics can resolve meaningful biochemical differences among soil systems under distinct land management practices. Methods: Soils from six land-use types—conventional cultivation, organic cultivation, pasture, white pine, tulip poplar, and hardwood forest—were analyzed using ultra-high-performance liquid chromatography coupled with high-resolution mass spectrometry (UHPLC-HRMS). Multivariate analyses, including PLS-DA, were performed to evaluate metabolic variation across systems. Both identified metabolites and unknown spectral features (MSI Level 4) were assessed, and biosynthetic class assignment of unknown features was performed using NPClassifier. Results: Metabolic features revealed clear separation between land management systems, demonstrating distinct chemical fingerprints across ecosystems. While conventional elemental ratios (e.g., C/N) showed minimal differentiation, phosphorus-related stoichiometric ratios (C/P and N/P) displayed strong land-use-dependent differences. NPClassifier superclasses highlighted unique chemical patterns, with forest soils enriched in diverse secondary metabolites, cultivated soils characterized by simplified profiles, and pasture soils dominated by microbial membrane lipids and alkaloids. Conclusions: Untargeted MS¹-based metabolomics effectively distinguished soil systems under different land-use practices and revealed ecologically meaningful variation even without complete structural identification. This study demonstrates that an MS¹-only workflow leveraging unknown spectral features can robustly distinguish soil systems, underscoring their value in untargeted metabolomics analyses. Full article

A growing number of studies evaluated cornelian cherry (Cornus mas L.) fruits due to their rich phenolic profile. It is obvious that the efficient recovery of these compounds depends on the extraction method used. Thus, this study aimed at comparing conventional (maceration, decoction) and non-conventional (ultrasound-assisted [UAE], enzyme-assisted [EAE]) extractions. Two novel aspects were characterized: the first application of EAE for cornelian cherry fruit phenolic recovery and the use of untargeted UHPLC-HRMS metabolomics. Untargeted UHPLC-HRMS profiling in positive electrospray ionization mode annotated 342 compounds, the broadest chemical analysis of C. mas fruits reported to date, followed by a semi-quantitative assessment of main phenolic compound subclasses using representative standards and multivariate statistical approaches. UAE proved most effective for recovering total flavonoids (433.6 μg eq./g) and phenolic acids (420.2 μg eq./g), while EAE selectively enriched other phenolic category compounds, semi-quantified as oleuropein equivalents (1975.4 μg eq./g). Multivariate analysis further confirmed distinct phytochemical fingerprints across extraction methods: the UAE extract was characterized by isocoumarin 2,4-di-O-methylolivetonide, while two compounds, cardamonin and ellagic acid arabinoside, were specifically up-accumulated in the EAE extract. All extracts showed moderate antioxidant (UAE > decoction > maceration > EAE) and enzyme inhibitory activities, with UAE and maceration being the most promising for multifunctional bioactivity. However, while untargeted UHPLC-HRMS revealed unprecedented complexity, results remain tentative and highlight the need for future targeted validation. Full article

Antarctic algae have evolved in extreme environmental conditions, developing unique metabolic adaptations with significant biotechnological potential. In this study, we explored the bioactivity of the sea-ice microalga Microglena antarctica by preparing acetone and methanol extracts from biomass cultivated at 4, 8, and 16 °C. These extracts were screened for their in vitro antioxidant properties and inhibitory activities on enzymes related to Alzheimer’s disease (acetylcholinesterase: AChE, butyrylcholinesterase: BChE), type 2 diabetes mellitus (T2DM, α-glucosidase, α-amylase), obesity (lipase), and hyperpigmentation (tyrosinase). Our screening revealed a high capacity of acetone extracts to scavenge the ABTS•⁺ radical (EC₅₀ ranging from 3.57 to 4.18 mg mL⁻¹), along with strong copper chelating activity in both acetone and methanol extracts (EC₅₀ values of 6.31 and 6.41 mg mL⁻¹). Relevant inhibition towards α-amylase (IC₅₀ values of 3.34 and 4.53 mg mL⁻¹) and tyrosinase (with IC₅₀ ranging from 3.82 to 5.47 mg mL⁻¹) was reported for acetone and methanol extracts, respectively. UHPLC-HRMS-based profiling revealed the presence of lipidic molecules, such as glycolipids, phospholipids, and betaine lipids with polyunsaturated carbon chains, together with carotenoids, including canthaxanthin and adonixanthin, which are likely responsible for the observed bioactivities. Full article

Plants are a valuable source of natural compounds with diverse applications. Recently, increased attention has focused on waste products from the agricultural industry, including walnut husk. Given its potential as a sustainable source of bioactives, this work characterizes the alcoholic Juglans regia “Sorrento” walnut husk extract (WHE). The extract’s phenolic content, antioxidant activity, and phytochemical composition were evaluated using spectrophotometry and UHPLC-HRMS-based untargeted metabolomics analysis. WHE exhibited a high total phenolic content (TPC = 1.45 ± 0.03 mg GAE/g dry extract) and a rich profile of phenolic acids, flavonoids, and tannins. Given this composition, WHE’s biological activity was further tested in an in vitro human keratinocyte (HaCaT) model. At the concentration of 10 μg/mL, WHE showed no cytotoxicity and displayed significant antioxidant properties by modulating detoxifying proteins such as Nrf2. WHE also influenced mitochondrial metabolism, increased maximum respiration, preserved barrier integrity, and activated pathways for epithelial homeostasis. Overall, this study highlights the bioactivity of the J. regia “Sorrento” walnut husk extract. These findings support the valorization of walnut husk as a sustainable source of bioactives for dermatological and cosmetic products. Full article

Feijoa (Acca sellowiana Berg) is an emerging Mediterranean crop valued for its nutraceutical potential but still underexplored with respect to cultivar and harvest stage. This study investigated two cultivars, ‘Mammoth’ and ‘Apollo’, harvested one week apart (4 and 11 November), to assess morphological traits, phenolic composition, antioxidant activity, vitamin C, and iodine. Fruit morphology, firmness, and basic quality indices (TSS, TA, pH, TSS/TA) were determined, while phenolic compounds were profiled by UHPLC–Q-Orbitrap HRMS. Antioxidant activity was measured by ABTS, DPPH, and FRAP assays; vitamin C by DCPIP titration; and iodine by iodometric analysis. ‘Apollo’ produced larger and firmer fruits, especially at the first harvest (105.6 g), while ‘Mammoth’ showed smaller and softer fruits. TSS remained stable (11 °Brix), whereas TA decreased and pH increased over time, raising the TSS/TA ratio and suggesting improved flavor balance at later harvests. Peel consistently contained higher bioactive levels than pulp, with catechin as the dominant phenolic compounds (up to 345 µg g⁻¹ dw in ‘Apollo’ peel). Antioxidant activity was markedly higher in peel, with ‘Mammoth’ showing stronger early FRAP values and ‘Apollo’ increasing at the later harvest. Vitamin C and iodine were about threefold higher in peel than pulp and increased over time, reaching maxima in late-harvest peel samples. Overall, cultivar and harvest stage significantly influenced fruit quality and nutraceutical value. Peel, particularly that of late-harvested ‘Apollo’, represents a promising resource for functional foods and the valorization of processing by-products. Full article

The ongoing challenge of doping in sports has triggered the adoption of advanced scientific strategies for the detection and prevention of doping abuse. This review examines the potential of integrating metabolomics aided by artificial intelligence (AI) and machine learning (ML) for profiling small-molecule metabolites across biological systems to advance anti-doping efforts. While traditional targeted detection methods serve a primarily forensic role—providing legally defensible evidence by directly identifying prohibited substances—metabolomics offers complementary insights by revealing both exogenous compounds and endogenous physiological alterations that may persist beyond direct drug detection windows, rather than serving as an alternative to routine forensic testing. High-throughput platforms such as UHPLC-HRMS and NMR, coupled with targeted and untargeted metabolomic workflows, can provide comprehensive datasets that help discriminate between doped and clean athlete profiles. However, the complexity and dimensionality of these datasets necessitate sophisticated computational tools. ML algorithms, including supervised models like XGBoost and multi-layer perceptrons, and unsupervised methods such as clustering and dimensionality reduction, enable robust pattern recognition, classification, and anomaly detection. These approaches enhance both the sensitivity and specificity of diagnostic screening and optimize resource allocation. Case studies illustrate the value of integrating metabolomics and ML—for example, detecting recombinant human erythropoietin (r-HuEPO) use via indirect blood markers and uncovering testosterone and corticosteroid abuse with extended detection windows. Future progress will rely on interdisciplinary collaboration, open-access data infrastructure, and continuous methodological innovation to fully realize the complementary role of these technologies in supporting fair play and athlete well-being. Full article

Despite the availability of effective vaccines, yellow fever outbreaks persist, highlighting the need for antiviral drugs. Background/Objectives: This study investigated Hippeastrum puniceum (Amaryllidaceae) as a potential source of antiviral compounds against wild-type yellow fever virus (wt-YFV). Methods/Results: The crude bulb extract of H. puniceum exhibited 58% protection against wt-YFV. Bioassay-guided fractionation of the extract by UHPLC-HRMS led to the annotation of six alkaloids (bulbisine, cathinone, trigonelline, tetrahydroharman-3-carboxylic acid, and 2,7-dimethoxyhomolycorine or 3-O-acetylnarcissidine) in active fractions, along with the amino acids arginine, asparagine, tryptophan, and glutamic acid. In silico ADMET analyses predicted favorable pharmacokinetic and toxicological profiles, supporting their potential as drug candidates. Six of the annotated compounds were evaluated in vitro for cytotoxicity and antiviral activity against wt-YFV. However, none showed significant antiviral activity when tested individually, suggesting that the observed antiviral effect may result from synergistic interactions between two or more compounds within active fractions. Conclusions: Our results underscore the importance of further investigations in vitro, particularly assays exploring the synergy among the annotated compounds against YFV. The integration of bioassay-guided fractionation of active plant extracts with computational analyses emerges as a promising strategy for the discovery of natural products with therapeutic potential against yellow fever, a reemerging disease. Full article

Nitrosamines (NAs) pose a risk due to their carcinogenic properties, especially in processed and cured meats where nitrites and nitrates are widely used. The objective of this study was to develop an integrated Ultra-High-Performance Liquid Chromatography–High-Resolution Mass Spectrometry (UHPLC–HRMS) workflow for detecting both volatile (VNAs) and non-volatile (NVNAs) nitrosamines in meat matrices. Comparison of two ionization techniques showed that heated electrospray ionization (HESI) and atmospheric pressure chemical ionization (APCI) provided complementary coverage and sensitivity. Extraction and cleanup were optimized for meat, although recovery rates remained variable, underscoring the analytical complexity. The method was applied to raw, cooked, cured, and grilled meats, as well as to in vitro gastric digestion and co-digestion with spinach. Results revealed that some NAs were present even in untreated raw meat (≈3.0 µg/kg, N-nitrosodi-n-butylamine), while the addition of nitrites and nitrates significantly increased their levels (more than 10 µg/kg, N-nitrosodiethylamine, N-nitrosodimethylamine, N-nitrosodi-n-butylamine). Gastric digestion was the most critical condition, further promoting nitrosamine formation, particularly for N-nitrosodiethylamine, N-nitrosodi-n-butylamine, and N-nitrosopiperidine. Ascorbate exhibited a dual role, acting as an inhibitor at low nitrite concentrations but becoming pro-oxidant at high levels (300 mg/kg). Cooking alone had limited impact, whereas cooking combined with digestion yielded the highest and most consistent nitrosamine concentrations. The inclusion of spinach during digestion modestly altered nitrosamine levels, reflecting both its nitrate content and polyphenolic profile. Nonparametric ANOVA (aligned rank transform) confirmed that preservative treatment, rather than processing or interaction effects, was the main driver of variability (total nitrosamines: H = 24.15, p = 2.33 × 10⁻⁵), with the combination of preservative ascorbate plus nitrite producing significantly higher levels than other treatments (q = 0.000656). N-nitrosodimethylamine consistently emerged as the most relevant marker for dietary exposure, in agreement with EFSA guidance. Overall, this study underscores both the analytical and biochemical complexity of nitrosamine detection and formation in meat products, while highlighting the importance of preservative formulation and the potential role of dietary antioxidants in mitigating exposure. Full article

Background: Bile salt hydrolase (BSH) is a key probiotic trait, as it facilitates both host metabolism and bacterial survival into the gastrointestinal tract (GIT), through bile acid (BA) deconjugation, keeping intestinal homeostasis. Objectives: The present study aims to investigate the viability of the Lacticaseibacillus rhamnosus VB4 strain and its effects on bile acid deconjugation during the gastrointestinal tract (GIT) passage, under a fed condition, using the in vitro SHIME^® (Simulator of the Human Intestinal Microbial Ecosystem) model. Methods: Gastric, small intestinal and colonic fractions were monitored and a fecal slurry from a healthy donor was inoculated into the colonic compartment to establish the intestinal microbiota. Samples were collected at the end of stomach, duodenum, jejunum, ileum phases, and colon after 0, 16 and 24 h. Strain survival was assessed by culturing method, and bsh gene expression was revealed by quantitative PCR (qPCR). In addition, UHPLC/HR-MS was performed to reveal the hypothetical changes in BAs profile after strain administration. Results: Good survivability of the VB4 strain in the upper GIT was revealed. Furthermore, VB4-inculated sample showed sustained expression of bsh in both the stomach/small intestine and colon fractions at all sampling times. Analysis of the BAs profile shown that the VB4 strain reduced the levels of the main conjugated BAs in the small intestine under fed condition and improved the deconjugation efficiency during colonic transit compared with the control. Conclusions: These findings highlight the survivability of L. rhamnosus VB4 strain inside the gut and its potential as biotherapeutic BAs-mediator candidate, demonstrating that transcriptomic and metabolomic approaches coupled to a dynamic in vitro gut model represent a robust tool for selection of a BSH-positive probiotic candidate. Full article

The Pulsed Electric Field (PEF) technique represents a modern technology for treating and processing food and agricultural raw materials. The application of high-voltage electric pulses has been shown to modify macrostructure, improve extractability, and enhance the microbial safety of the treated matrix. In this study, we investigated metabolomic changes occurring during the individual technological steps of malting following PEF treatment. Methanolic extracts of technological intermediates of malting barley were analyzed using metabolomic fingerprinting performed with UHPLC-HRMS/MS. For data processing and interpretation, the freely available MS-DIAL—MS-CleanR—MS-Finder software platform was used. The metabolomes of the treated and untreated barley samples revealed significant changes. Tentatively identified PEF-related biomarkers included 1,2-diacylglycerol-3-phosphates, triacylglycerols, linoleic acids and their derivatives, octadecanoids, N-acylserotonins, and very long-chain fatty acids, and probably reflect abiotic stress response. Monitoring of the profiles of selected biomarkers in PEF malting batch indirectly revealed a potential enhancement of enzymatic activity after the PEF treatment. These results contribute to fundamental knowledge regarding the influence of PEF on final malt from a metabolomic perspective. 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