Search Results (383)

Background/Objectives: Possessing red and white ecotypes, and utilized in traditional Guyanese medicine, Doliocarpus dentatus’ red ecotype is preferred locally for its purported superior therapeutic efficacy. Although therapeutic metabolites were detected in D. dentatus previously, phytohormones remain largely unexplored, until now. Cytokinins, phytohormones responsible for plant cell division, growth and differentiation, are gaining traction for their therapeutic potential in human health. This study screened and quantified endogenous cytokinins and correlated detected cytokinins with selected secondary metabolites. Methods: Liquid chromatography–mass spectrometry was used to acquire phytohormone and metabolite data. Bioinformatics tools were used to assess untargeted metabolomics datasets via statistical and pathway analyses, and chemical groupings of putative metabolites. Results: In total, 20 of the 35 phytohormones were detected and quantified in both ecotypes, with the red ecotype displaying higher free base and glucoside cytokinin concentrations and exhibited 6.2 times the total CK content when compared to the white ecotype. Pathway analysis revealed flavonoid and monoterpenoid biosynthesis in red and white ecotypes, respectively. Positive correlations between specific cytokinins and alkaloids, and between trans-Zeatin and isopentenyladenosine riboside with phenolic compounds were observed. Conclusions: These results suggest that the red ecotype’s elevated cytokinin levels coupled with flavonoid biosynthesis enrichment support its preference in Guyanese traditional medicine. Full article

Recombinant adeno-associated virus (rAAV) is the leading vector for gene replacement therapy; however, the roles and regulation of host proteins in rAAV production remain incompletely understood. In this comparative proteomic analysis, we focused on proteins in the nucleus, the epicenter of DNA uptake, transcription, capsid assembly, and packaging. HEK-293 cells were analyzed under the following three conditions: (i) untransfected, (ii) mock-transfected with the ITR and an unrelated plasmid, and (iii) triple-transfected with rAAV2 production plasmids. Cells were harvested at 24 and 72 h post-transfection, and nuclear fractions were processed using filter-aided sample preparation (FASP) followed by nano-scale liquid chromatography–tandem mass spectrometry (nLC-Orbitrap MS/MS). Across all samples, we identified 3384 proteins, revealing significant regulatory changes associated with transfection and rAAV production. Transfection alone accounted for some of the most substantial proteomic shifts, while rAAV production induced diverse regulatory changes linked to cell cycle control, structure, and metabolism. STRING analysis of significantly regulated proteins also identified an enrichment of those associated with the Gene Ontology (GO) term ‘response to virus’. Additionally, we examined proteins with reported relation to adenoviral components. Our findings help to unravel the complexity of rAAV production, identify interesting targets for further investigation, and may contribute to improving rAAV yield. Full article

This study explores the identification, characterization, and biological evaluation of angiotensin I-converting enzyme (ACE)-inhibitory peptides derived from enzymatic hydrolysates of crucian carp swim bladders. Following sequential purification by size-exclusion and reversed-phase chromatography, two bioactive peptides—Hyp-Gly-Ala-Arg (Hyp-GAR) and Gly-Ala-Hyp-Gly-Ala-Arg (GA-Hyp-GAR)—were identified using ultra-high-performance liquid chromatography coupled with linear ion trap–Orbitrap tandem mass spectrometry. The synthetic peptides demonstrated potent ACE-inhibitory activity in vitro, with IC₅₀ values of 12.2 μM (Hyp-GAR) and 4.00 μM (GA-Hyp-GAR). Molecular docking and enzyme kinetics confirmed competitive inhibition through key interactions with ACE active site residues and zinc coordination. In vivo antihypertensive activity was evaluated in spontaneously hypertensive rats, revealing that GA-Hyp-GAR significantly reduced systolic blood pressure in a dose-dependent manner. At a dose of 36 mg/kg, GA-Hyp-GAR reduced systolic blood pressure by 60 mmHg—an effect comparable in magnitude and timing to that of captopril. Mechanistically, GA-Hyp-GAR modulated levels of angiotensin II, bradykinin, endothelial nitric oxide synthase, and nitric oxide. A 90-day subchronic oral toxicity study in mice indicated no significant hematological, biochemical, or histopathological alterations, supporting the peptide’s safety profile. These findings suggest that GA-Hyp-GAR is a promising natural ACE inhibitor with potential application in functional foods or as a nutraceutical for hypertension management. Full article

This study investigates the compositional characteristics and quantitative differences of ginsenosides across various ginseng tissues, with a particular focus on the specific accumulation patterns of malonyl ginsenosides. Five tissue samples—ginseng fruit (F), leaf (L), taproot (TR), lateral root (LR), and fibrous root (FR)—were analyzed using Orbitrap Fusion high-resolution liquid chromatography–mass spectrometry. A total of 413 ginsenosides, including 33 standards, were identified, encompassing 172 protopanaxadiol (PPD)-type, 188 protopanaxatriol (PPT)-type, 14 oleanolic acid (OA)-type, and 12 ocotillol (OT)-type ginsenosides, of which 160 were malonyl ginsenosides. Statistical analysis revealed significant variations in the relative content per unit mass of malonyl ginsenosides across tissues, with the highest levels in fibrous roots, followed by fruits, lateral roots, leaves, and taproots. Distinct tissue-specific differences in malonyl ginsenoside types and quantities were observed: fruits exhibited 51 malonyl ginsenosides with significantly higher levels, compared to 8, 14, and 17 in lateral roots, fibrous roots, and leaves, whereas TR showed no significant enrichment. This study elucidates the diversity and unique distribution of malonyl ginsenosides in ginseng roots, leaves, and fruits, providing a valuable basis for the targeted selection of tissues with high malonyl ginsenoside content and the development of functional food and medicinal products. Full article

Organophosphorus (OP) pesticides are a class of chemicals that are extensively used worldwide. The exposure to and use of organophosphates can be assessed by analyzing their metabolites and degradation products, such as dialkyl phosphate (DAP), dialkyl thiophosphate (DATP), and dialkyl dithiophosphate (DADTP). However, since these metabolites/hydrolysis products can result from the metabolism or breakdown of several organophosphorus pesticide families, they serve as nonspecific biomarkers and do not indicate the specific pesticide involved in exposure. In an earlier study, chemical derivatization using N-(2-(bromomethyl)benzyl)-N,N-diethylethanaminium bromide (CAX-B) was described to improve the signal intensity of numerous organophosphorus (OP) acids in liquid chromatography tandem mass spectrometry (LC-ESI-MS/MS) analysis. In the present study, CAX-B was employed to derivatize a set of seven phenolic compounds corresponding to the complementary portion of OP pesticides. The derivatization process using CAX-B was performed in acetonitrile with potassium carbonate at 50 °C for 30 min. LC-Orbitrap-ESI-MS/MS was used to analyze the resulting phenol derivatives and their fragmentation patterns were studied. Notably, the derivatized phenols were markedly more sensitive than the underivatized phenols when LC-ESI-MS/MS was used in MRM technique, without being affected by the sample matrix (soil or plant extracts). This derivatization technique aids in identifying OP pesticides, offers insights into their subfamily, and pinpoints a specific compound through the analysis of corresponding phenol derivative. Full article

The degradation of pharmaceuticals in wastewater treatment plants (WWTPs), particularly the antidepressant Paroxetine (PXT), is a growing concern because their insufficient removal leads to their release in the aquatic environment, causing toxic effects on aquatic organisms. This study investigates g-C₃N₄ materials synthesized from urea, melamine, and thiourea, including thermally exfoliated variants, as potential photocatalysts for removing PXT from water and secondary-treated hospital wastewater (HWW). Comparative photocatalytic experiments under simulated solar radiation indicated that g-C₃N₄ prepared by urea (CN-U) and its thermally exfoliated form [CN-U(exf.)] were highly effective (100% removal in 45 min) depending on the degradation rate constants (0.036 and 0.085 min⁻¹ in U.P. water, respectively), with the latter achieving the fastest PXT degradation at 200 mg/L (k = 0.112 min⁻¹). The study also analyzed mineralization and transformation products (TPs) using liquid chromatography–high-resolution mass spectrometry (LC–HR-MS-Orbitrap) and assessed their ecotoxicity with ECOSAR (Version 2.2) software. Additionally, toxicity decreased following the photocatalytic processes, as revealed by the Microtox bioassay. Overall, CN-U and especially CN-U(exf.) show promise as eco-friendly photocatalysts for pharmaceutical removal from wastewater (WW). Full article

Background: Chronic atrophic gastritis precancerous lesions (PL-CAG) are characterized by the atrophy of gastric mucosal glands, often accompanied by intestinal metaplasia or dysplasia. Timely intervention and treatment can effectively reverse its malignant progression and prevent the onset of gastric cancer. Bombyx Batryticatus (BB) exhibits a range of pharmacological effects, including anticoagulation, antiepileptic properties, anticancer activity, and antibacterial effects. However, the pharmacological basis and mechanisms underlying BB’s efficacy in treating PL-CAG remain unclear. Methods: A three-factor modeling approach was implemented to develop a rat PL-CAG model, while the MNNG-induced PLGC (precancerous lesions of gastric cancer) cell model was served as a cell PL-CAG model. UPLC-QE-Orbitrap-MS/MS (Ultra performance liquid chromatography-quadrupole-electrostatic field orbital trap high-resolution mass spectrometry) was utilized to perform an in-depth analysis of the components in the plasma extract of BB. Leveraging network pharmacology, molecular docking analyses, and experimental validation, we initially elucidated the potential mechanisms through which BB mediates its therapeutic effects on PL-CAG at both in vivo and in vitro levels. Results: Prototype compounds of 42 blood-entering components were identified by UPLC-QE-Orbitrap-MS/MS analysis. Network pharmacology analysis and molecular docking studies indicate that the core targets are primarily enriched in the PI3K-Akt signaling pathway, and the key components, including Nepitrin, Quercetin 3-O-neohesperidoside, Rutin, and others, exhibited stable docking conformations with the first eleven pivotal targets. Both in vivo and in vitro experiments validated that BB may effectively treat PL-CAG via modulation of the PI3K-Akt signaling pathway. Conclusions: The therapeutic efficacy of BB in the management of PL-CAG may be achieved through the synergistic interaction of multiple components and targets, which may be more closely related to the inhibition of the PI3K/AKT signaling pathway. This approach will establish a solid experimental foundation and provide essential data for the clinical application of BB in treating PL-CAG, while also facilitating further research initiatives. Full article

In Romania, groundwater is an important source of drinking water, especially in rural areas. This study investigated the concentrations of organophosphorus, carbamate, and triazine pesticides (OPs) along with organochlorine pesticides (OCPs) in groundwater samples collected from the Titu-Sarata Plain. Sensitive analytical techniques were employed, including Ultrahigh-Performance Liquid Chromatography coupled with Q Exactive™ HF Hybrid Quadrupole-Orbitrap™ Mass Spectrometry (UHPLC-Orbitrap-MS/MS) and Gas Chromatography coupled with an electron capture detector (GC-ECD). Environmental and human health risks were assessed in the case of pesticides that exceeded the maximum allowed concentration. The environmental risk assessment (ERA) revealed significant risks associated with Phosdrin, Phorate, and pp’DDE. Additionally, particular concerns arose from the presence of Aldrin and Dieldrin, which pose a high carcinogenic risk, especially through groundwater consumption in agricultural areas. The results of this research highlight the need for the implementation of a continuous quality monitoring program for groundwater in the agricultural regions that were studied. Full article

Throughout the process of dry-cured ham, moulds such as P. nordicum, a producer of ochratoxin A (OTA), grow on its surface. The use of combined biocontrol agents (BCAs) is a promising strategy for controlling this hazard. The goal of this study is to assess the effect of D. hansenii, S. xylosus, and P. chrysogenum as BCAs on the metabolome of two strains of P. nordicum and to understand the differences between both strains. Each ochratoxigenic strain was inoculated both individually and in combination with the BCAs onto ham for 30 days under the environmental conditions experienced during traditional ripening. Untargeted metabolomics was performed through mass spectrometry using a Q-Exactive Plus Orbitrap. The BCAs caused alterations in the metabolomes of both ochratoxigenic moulds, mainly in phenylalanine catabolism and the valine, leucine, and isoleucine biosynthesis pathways, although with some differences. In the absence of the BCAs, the metabolomes of both types of P. nordicum were globally changed, despite these being moulds of the same species. In conclusion, these data help us to understand the differences between OTA-producing strains in dry-cured ham and confirm the need to demonstrate the efficacy of BCAs against a wide range of toxigenic moulds before they can be used to minimise OTA contamination in the meat industry. Full article

Yinhua Pinggan Granules (YPG) is a patented traditional Chinese medicine (TCM) compound prescription, with wide clinical application against cold, cough, and relevant diseases. However, the chemical profiles of YPG in vivo are still unknown, hindering further pharmacological and quality control (QC) researches. This study presents an ultra-high-performance liquid chromatography coupled with high-resolution orbitrap mass spectrometry (UHPLC-MS)-based method. Using the Compound Discoverer platform and a self-built ‘in-house’ compound database, the metabolic profiles and pharmacokinetics characters of YPG were investigated. Consequently, a total of 230 compounds (including 39 prototype components and 191 metabolites) were tentatively identified, in which the parent compounds were mainly flavonoids, alkaloids, and terpenoids, and the main metabolic pathways of metabolites include hydration, dehydration, and oxidation. The serum concentration of seven major representative compounds, including quinic acid, chlorogenic acid, amygdalin, 3′-methoxypuerarin, puerarin, glycyrrhizic acid, and polydatin, were also measured, to elucidate their pharmacokinetics behaviors in vivo. The pharmacokinetic study showed that the seven representative compounds were quantified in rat plasma within 5 min post-administration, with T_max of less than 2 h, followed by a gradual decline in concentration over a 10 h period. The method demonstrated excellent linearity (R² > 0.998), precision, and recovery (RSD < 15%). As the first systematic characterization of YPG’ s in vivo components and metabolites using UHPLC-MS, this study may contribute to comprehensively elucidate the metabolic profiles of the major components in YPG, and provide a critical foundation for further investigation on the QC and bioactivity research of YPG. Full article

We investigated the dynamic changes in volatile aroma compound profiles (types and concentrations) and associated gene expression patterns in both the peel and pulp tissues of apples during fruit maturation. This study aimed to elucidate the metabolic regulatory mechanisms underlying volatile aroma biosynthesis in Malus domestica “Ningqiu” apples, thereby providing theoretical support for the comprehensive utilization of aroma resources. Our methodological framework integrated headspace solid-phase microextraction gas chromatography–mass spectrometry (HS-SPME-GC-MS), ultra-high-performance liquid chromatography–orbitrap mass spectrometry (UHPLC-OE-MS), and Illumina high-throughput sequencing to generate comprehensive metabolomic and transcriptomic profiles of peel and pulp tissues. Critical differential aroma compound classes were identified, including esters, aldehydes, alcohols, terpenoids, and ketones, with their metabolic pathways systematically mapped through KEGG functional annotation. Our findings revealed substantial transcriptomic and metabolomic divergence across carotenoid, terpenoid, and fatty acid metabolic pathways. Integrative analysis of multi-omics data revealed 26 and 31 putative biologically significant hub genes in peel and pulp tissues, respectively, putatively associated with the observed metabolic signatures. Among these, five core genes—farnesyl diphosphate synthase (FDPS1.X1), alcohol acyltransferases (AAT1 and AAT3), alcohol dehydrogenase (ADH3), and carotenoid cleavage dioxygenase (CCD3)—were recognized as shared regulatory determinants between both tissue types. Furthermore, terpene synthase (TPS7) emerged as a peel-specific regulatory factor, while hydroperoxide lyase (HPL2), alcohol dehydrogenases (ADH2 and ADH4), and alcohol acyltransferase (AAT2) were identified as pulp-exclusive modulators of metabolic differentiation. The experimental findings provide foundational insights into the molecular basis of aroma profile variation in Malus domestica “Ningqiu” and establish a functional genomics framework for precision breeding initiatives targeting fruit quality optimization through transcriptional regulatory network manipulation. Full article

Babesiosis is a significant vector-borne zoonotic disease with major global economic and health implications, affecting various vertebrate hosts. Babesia parasites are auxotrophic for essential nutrients, relying on their hosts for metabolic support. This study investigated salivary metabolomic changes in dogs naturally infected with Babesia canis (N = 14) compared to healthy controls (N = 14) using untargeted and targeted mass spectrometry-based approaches. Saliva, a biofluid rich in metabolites, undergoes alterations in response to systemic diseases, making it a promising medium for studying host–pathogen interactions. Metabolomic profiling was performed using a Dionex UltiMate 3000 UHPLC system coupled with a Thermo Orbitrap Q Exactive mass spectrometer. An untargeted analysis detected 2257 salivary features, of which, 868 were significantly altered, with seven metabolites identified by reference to standards. A targeted analysis revealed significant changes in seven metabolites. Functional bioinformatics indicated disruptions in amino acid, nucleotide, and lipid metabolism, alongside alterations in energy production pathways, and purine metabolism. These findings provide critical insights into the metabolic shifts underlying canine babesiosis, supporting the development of advanced diagnostic and therapeutic strategies in the future. This study highlights the intricate interplay between host and pathogen, particularly in nutrient acquisition and metabolic regulation. Full article

Coenzyme Q10 (CoQ10), a high-value-added nutraceutical antioxidant, exhibits an excellent ability to prevent cardiovascular disease. Here, a novel Cereibacter azotoformans strain, designated YS02, was isolated for its ability to produce CoQ10 and genetically characterized by whole genome sequencing (WGS). The CoQ10 biosynthesis and metabolism differences of YS02 under various culture conditions were also systematically investigated. Phylogenetic analysis based on 16 S rRNA genes, along with taxonomic verification using average nucleotide identity (ANI) analysis, confirmed its classification as C. azotoformans. Enzymatic genes dxs, dxr, idi, ubiA, and ubiG were annotated in YS02, which are critical genetic hallmarks for CoQ10 biosynthesis. Under aerobic–dark cultivation, YS02 grows well, and CoQ10 production can reach 201 mg/kg. A total of 542 small-molecule metabolites were identified from YS02 in aerobic–dark and anaerobic–light cultivation via ultra-high performance liquid chromatography–coupled quadrupole orbitrap high-resolution mass spectrometry (UPLC-Q-Exactive Orbitrap MS). Additionally, 40 differential metabolites were screened through multivariate statistical analysis. Metabolic pathway analysis revealed that the biosynthesis of phenylalanine, tyrosine, and tryptophan might be latent factors influencing CoQ10 production discrepancies within YS02 under both cultural modes. These findings represent new insights into the metabolic mechanism of YS02 and underscore its potential as an alternative strain source for industrial CoQ10 production, enriching the existing resources. Full article

Gelatin from deer has garnered attention as a high-value health-promoting resource given its history of usage as a traditional Chinese medicine and recent studies demonstrating its biological activities. Mass spectrometry-based methods have increasingly been employed for species identification in collagen-based materials, effectively addressing challenges in quality control and authenticity verification. This study aims to identify characteristic marker peptides in gelatins from sika deer (Cervus nippon) to support their effective use as a health-promoting resource. Gelatin samples were enzymatically digested, and the resulting peptide mixtures were analyzed using ultra-high-performance liquid chromatography coupled with quadrupole Q-Exactive-Orbitrap mass spectrometry (UHPLC-Q-Exactive-Orbitrap MS). Marker peptide candidates were selected based on their high detection intensity and a literature review. Among the 28 selected marker peptide candidates, four peptides (P11, R2, R3, and R4) were defined as characteristic of sika deer gelatin. Comparative analyses with gelatins derived from donkey hide, bovine, porcine, and fish samples further confirmed the specificity of these peptides. These findings establish a robust analytical method for verifying the authenticity of sika deer gelatin, contributing to its safe and effective use as a health-promoting resource. 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