Search Results (4,302)

Cyanobacterial harmful algal blooms (cyanoHABs) typically involve the release of a wide spectrum of toxic metabolites produced by different cyanobacteria, which can cause adverse health effects in animals and humans. At the same time, many cyanobacteria are capable of producing more than one toxin simultaneously. As a result, the multiclass toxins released into aquatic environments can exert complex and combined effects on various ecosystem components. This review aims to provide an overview of the occurrence and effects of different cyanotoxins in aquatic systems and current analytical methods of their simultaneous detection. The main classes of cyanotoxins have been considered in this review. The occurrences, properties, and environmental and health impacts of these toxins are described. The main methods for cyanobacterial toxin analysis are covered. A comprehensive review and analysis of liquid chromatography-tandem mass spectrometry (LC-MS/MS) detection and sample preparation approaches used to identify various classes of cyanotoxins is provided, together with a practical decision tree for analytical workflow selection according to toxin class and sample matrix, highlighting key trends and outlining directions for further research in this area. Full article

Background/Objectives: Tropane alkaloids (TAs) and phenylpropanoid polyamine alkaloids (PPAs) are major alkaloid constituents of solanaceous plants; however, the tissue-specific distribution of these compounds in Anisodus luridus, a medicinal plant native to southeastern Xizang, remains incompletely characterized. Methods: A biosynthesis-informed putative annotation workflow was developed using ultra-performance liquid chromatography–high-resolution mass spectrometry (UPLC-HRMS) to characterize alkaloid profiles across seven tissues (root, stem, leaf, calyx, corolla, ovary, and seed). Putative annotations were guided by substrate–product structural relationships, predicted enzyme-catalyzed modifications, and diagnostic tandem mass spectrometry (MS/MS) fragmentation patterns. Results: Eighteen alkaloid features were characterized, comprising 6 TAs and 12 PPAs. Hyoscyamine and scopolamine were identified using authentic standards, and the remaining 16 features were putatively annotated, including dihydroanisodamine, hexosylhyoscyamine, and several PPAs not previously reported in A. luridus. TAs were predominantly enriched in roots and ovaries, whereas PPAs accumulated in floral tissues and leaves. Orthogonal partial least squares discriminant analysis (OPLS-DA) identified scopolamine (variable importance in projection (VIP) = 1.30) as a principal variable associated with separation among tissue metabolic profiles. Conclusions: This workflow supports the putative annotation of alkaloids in non-model medicinal plants. The observed tissue-specific distribution patterns provide a basis for future comparative studies with A. belladonna and for evaluating cultivation, tissue-selective harvesting, and extraction strategies. Full article

Background: The administration of direct oral anticoagulants to older adults requires careful risk stratification against a backdrop of polypharmacy and age-related renal function decline. The role of single nucleotide polymorphisms in cytochrome P450 genes and efflux transporters in this context remains a clinically understudied issue. This study aimed to investigate the association of ABCB1, CYP3A4, and CYP3A5 allelic variants with the safety of rivaroxaban therapy in geriatric patients with non-valvular atrial fibrillation (AF). Materials and Methods: This prospective cohort study consecutively enrolled 94 patients (mean age 83.2 ± 9.2 years). Rivaroxaban trough plasma concentrations ($C_{min,ss}$ ) were quantified using a validated high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) assay. Real-time polymerase chain reaction was utilized for the molecular genetic profiling of the ABCB1 (rs1045642, rs2032582), CYP3A4*22 (rs35599367), and CYP3A5*3 (rs776746) loci. Comorbidity severity and the spectrum of drug–drug interactions were evaluated as additional predictors. Results: Clinically relevant hemorrhagic complications were documented in 36.2% (n = 34) of the monitored patients. A significant association was established between the homozygous CC genotype at the ABCB1 rs1045642 locus and increased bleeding propensity (odds ratio [OR] 2.42; 95% CI: 1.02–5.74; p = 0.042), whereas the alternative TT variant was associated with a pronounced protective effect. No statistically significant correlations were identified for biotransformation isoenzyme markers (CYP3A4*22, CYP3A5*3). Anticoagulant metrics were comparably distributed across groups and showed no dependence on genetic status. The leading non-genetic factors associated with hemorrhage were advanced age (p < 0.0001), progressive glomerular filtration rate decline (CKD stages 3b–4; p = 0.0009), and pharmacokinetic/pharmacodynamic interference from the co-administration of amiodarone (OR 3.61), non-steroidal anti-inflammatory drugs, and antiplatelet agents. The validated HAS-BLED score demonstrated no predictive power within the comorbid conditions of this cohort (p = 0.052). Conclusions: The ABCB1 rs1045642 (C3435T) polymorphism is associated with the occurrence of hemorrhagic events. Implementing pharmacogenetic testing of the P-glycoprotein transporter combined with a rigorous audit of renal excretory function and concomitant therapy management may optimize the safety profile of rivaroxaban in geriatric practice. These findings warrant confirmation in a larger cohort. Full article

Ligusticum sinense Oliv. cv. Chaxiong (Chaxiong) has traditionally been used as a medicinal tea for headache relief in southern China, but its bioactive basis and mechanisms remain unclear. This study integrated ultra-performance liquid chromatography–quadrupole time-of-flight tandem mass spectrometry (UPLC-Q-TOF-MS/MS), untargeted metabolomics, network pharmacology, molecular docking, reverse-transcription quantitative PCR (RT-qPCR), and Western blotting in a nitroglycerin-induced migraine-like rat model. Chaxiong extract alleviated migraine-like behavior, reduced calcitonin gene-related peptide (CGRP), nitric oxide (NO), and tumor necrosis factor-α (TNF-α), increased 5-hydroxytryptamine (5-HT), and ameliorated brain histopathological alterations. A total of 48 constituents and 13 differential serum metabolites were annotated or putatively annotated, respectively. Glycerophospholipid metabolism was the principal metabolic pathway affected by treatment, whereas network pharmacology prioritized phosphatidylinositol 3-kinase/protein kinase B (PI3K/AKT) signaling. Pathway integration identified PIK3CA, PIK3CB, AKT1, AKT2, and INS as candidate bridging targets. Exploratory docking suggested potential interactions between five representative constituents and PIK3CA/AKT1. RT-qPCR showed reduced Pik3ca and Akt1 mRNA expression, while Western blotting showed decreased p-PI3K p85α (Tyr607)/PI3K p85α and p-AKT (Ser473)/AKT1 ratios following treatment. These findings suggest that Chaxiong-associated alleviation of migraine-like responses was accompanied by altered glycerophospholipid metabolism and reduced PI3K/AKT pathway activation. Full article

Bile acid (BA) dysmetabolism is implicated in canine chronic inflammatory enteropathy (CIE), but fasting serum profiles across treatment-response groups remain poorly characterized. This exploratory observational study analyzed pretreatment serum from dogs subsequently classified as food-responsive enteropathy (FRE; n = 8) or immunosuppressant-responsive enteropathy (IRE; n = 5), with four unmatched purpose-bred Beagle controls. Targeted LC–MS/MS, performed using a published canine serum method, provided results for 15 BA species. All samples were analyzed in one batch by an analyst blinded to subsequent group assignment. Eight species quantifiable in every dog and ten derived variables were compared using Kruskal–Wallis tests with Benjamini–Hochberg correction across 18 omnibus tests. Cholic acid and unconjugated primary BA concentrations were higher in IRE than in controls and FRE. The total primary-to-secondary BA ratio showed a monotonic group ordering (control < FRE < IRE; H = 13.76, p = 0.0010, q = 0.0092; epsilon-squared = 0.840), with all Bonferroni-corrected pairwise comparisons significant. A post hoc four-scenario sensitivity analysis comprising the submitted zero-substitution analysis and three LOD/LOQ-based alternatives reproduced the same 18-test FDR-significant set and the same ratio inference. This analysis tests robustness to censored-value handling but is not independent validation. The findings suggest group-associated serum BA patterns, but the small sample, clinical heterogeneity, and unmatched controls preclude diagnostic or treatment-predictive interpretation. Full article

The gut microbiota plays a critical role in regulating liver metabolism and immunity, significantly influencing hepatic inflammation and fibrogenesis. However, the mechanisms by which it modulates the progression of liver fibrosis remain unclear. This study investigated the interplay between the gut microbiota and bile acids in murine liver fibrosis. A mouse model of liver fibrosis was established via intraperitoneal injection of carbon tetrachloride. The gut microbiota was depleted using an antibiotic cocktail, as confirmed through 16S rRNA sequencing. Bile acid profiles were measured using ultra-performance liquid chromatography–tandem mass spectrometry. The effects of secondary bile acids on macrophage polarization were assessed in vivo and in vitro. Gut commensal microbiota depletion exacerbated liver inflammation and fibrosis. Concomitantly, this depletion significantly reduced microbiota-derived secondary bile acids. Notably, supplementation with deoxycholic acid markedly attenuated liver fibrosis. This protective effect was associated with the inhibition of pro-inflammatory M1 macrophage polarization. Takeda G protein-coupled receptor 5 (TGR5) activation was identified as the mechanism underlying the effect of deoxycholic acid on macrophages, as a TGR5 antagonist reversed this inhibition. Mechanistically, TGR5 activation suppressed the NF-κB pathway in macrophages via the cAMP–PKA signaling cascade, thereby inhibiting hepatic stellate cell activation. Our results demonstrate that the gut microbiota–secondary bile acid–TGR5 signaling axis plays a critical role in liver fibrosis and represents a promising therapeutic target. Full article

Background/Objective: Accurate measurement of serum 25-hydroxyvitamin D (25OHD) is essential for the diagnosis and management of vitamin D deficiency. Although automated immunoassays are widely used in routine clinical practice, analytical variability between platforms may lead to inconsistent patient classification around clinically relevant decision thresholds. This study comprehensively compared three automated immunoassays (DiaSorin Liaison XL, Beckman Coulter DxI 9000, and Roche cobas e602) with a Vitamin D Standardization Certification Program (VDSCP)-traceable liquid chromatography–tandem mass spectrometry (LC-MS/MS) reference method. Methods: A total of 400 anonymized adult outpatient serum samples were prospectively collected and equally distributed across four predefined vitamin D concentration categories (<25, 25–49, 50–74, and ≥75 nmol/L). Samples were stored at −70 °C before simultaneous analysis on the three automated immunoassays and the reference LC-MS/MS platform. Method comparison included Deming regression, Bland–Altman analysis, and evaluation of diagnostic performance at clinically relevant thresholds (<25, <50, and <75 nmol/L). The effects of sample storage, vitamin D metabolites (C3-epi-25OHD₃ and 24,25-dihydroxyvitamin D₃), and external quality assessment (DEQAS) performance were also investigated. Results: Significant analytical differences were observed between all immunoassays and LC-MS/MS. Roche cobas e602 consistently overestimated 25OHD concentrations, resulting in reduced sensitivity but excellent specificity and positive predictive value across all clinical thresholds. Beckman Coulter demonstrated the highest sensitivity for detecting vitamin D deficiency but generated more false-positive classifications, particularly at the <75 nmol/L threshold. DiaSorin Liaison XL achieved the best overall balance between sensitivity, specificity, accuracy, and agreement with LC-MS/MS, with Cohen’s κ values ranging from 0.818 to 0.895 across decision thresholds. Higher concentrations of C3-epi-25OHD₃ and 24,25-dihydroxyvitamin D₃ were associated with greater positive bias in Roche measurements, while Beckman Coulter showed no significant metabolite-related interference, although the present correlation analyses could not establish independent analytical interference. DEQAS performance did not consistently reflect agreement with LC-MS/MS in patient samples. Conclusions: Despite ongoing international standardization efforts, clinically important differences remain among automated 25OHD immunoassays. Assay-specific analytical characteristics significantly influence patient classification around established decision thresholds. DiaSorin Liaison XL demonstrated the closest overall agreement with the reference LC-MS/MS method, whereas Beckman Coulter prioritized sensitivity and Roche specificity. These findings highlight the importance of method-aware interpretation of vitamin D results and support continued assay harmonization to improve comparability in routine clinical practice. Full article

Triclosan (TCS) and triclocarban (TCC) are antimicrobial compounds widely used in numerous industries, making them significant environmental pollutants that infiltrate living organisms and exhibit endocrine-disrupting activity. However, knowledge regarding the exposure of wild terrestrial mammals to these substances remains virtually non-existent. Therefore, this study analyzed, for the first time, TCS and TCC levels in the hair of wild boars using liquid chromatography-tandem mass spectrometry (LC-MS). Despite widespread environmental contamination with these compounds, concentrations above the limit of detection (LOD: 2.3 pg/mg for TCS and 0.8 pg/mg for TCC) were detected in only a small percentage of the analyzed samples: 14.81% for TCC and 3.70% for TCS. Higher concentrations and a greater frequency of TCC occurrence were observed in animals from highly urbanized and industrialized regions. In contrast, TCC and TCS concentrations in animals from other areas only sporadically exceeded the LOD. These results suggest that wild boars are primarily exposed to TCC in highly urbanized areas. In less-urbanized regions, animal exposure to both compounds is usually below the LOD and likely depends on poorly defined local factors. Nonetheless, fully clarifying the exposure of wild terrestrial mammals to TCC and TCS requires further comprehensive research. Full article

Bacillus velezensis is a promising candidate for the biocontrol of plant pathogens due to its production of broad-spectrum antimicrobial metabolites. The present study evaluates the antifungal potential and metabolic profile of the non-pathogenic rhizobacterial strain Bacillus velezensis 5RB. Modifying the mineral composition across three different growth media (A, B, and C) resulted in distinct secondary-metabolite profiles of B. velezensis 5RB. Notably, the free-cell supernatant of strain 5RB cultivated in medium A and C demonstrated the most potent antifungal activity against the gray mold pathogen Botrytis cinerea. For the first time, the antimicrobial metabolites produced by B. velezensis 5RB across these three media were comprehensively characterized using ultra-performance liquid chromatography-quantitative time-of-flight mass spectrometry (UHPLC-QTOF-MS). Mass spectrometric analyses (LC-ESI-MS) identified predominantly lipopeptides from the surfactin and fengycin families, alongside three classes of polyketides: macrolactins, difficidins, and bacillaenes. Multiple lipopeptide homologues, including a novel surfactin isoform (B*), were confirmed by tandem mass spectrometry (LC-ESI-MS/MS). Additionally, this study provides the first experimental evidence for subtilosin A production by B. velezensis 5RB. The multicomponent fermentation matrix of 5RB conferred robust biocontrol protection to tomato leaves against B. cinerea and Phytophthora infestans in an in vitro disease model, underscoring its strong potential for sustainable agricultural applications. Full article

Background/Objectives: Origanum bilgeri P.H. Davis is a Turkish endemic Lamiaceae species whose non-volatile phenolic composition and organ-specific bioactivities remain unexplored. This study investigated the phenolic composition of flower, leaf, stem, and root extracts and evaluated their antioxidant and enzyme-inhibitory activities. Methods: Ultrasound-assisted ethanolic extracts of the four organs were characterized by liquid chromatography–tandem mass spectrometry (LC–MS/MS) targeting 28 phenolic compounds. Total phenolic and flavonoid contents, outcomes of six antioxidant assays, and inhibitory activities against acetylcholinesterase, butyrylcholinesterase, tyrosinase, α-amylase, and α-glucosidase were determined. Measurements were performed in triplicate as technical replicates, and organ-level differences were interpreted descriptively. The relative antioxidant capacity index (RACI) was calculated from the standardized outcomes of the six direct antioxidant assays, with total phenolic content (TPC) and total flavonoid content (TFC) excluded from the index. Pearson correlation analysis and principal component analysis (PCA) were used for exploratory evaluation of phytochemical and bioactivity patterns. Results: Twenty phenolic compounds were detected in at least one organ. Rosmarinic acid predominated in all organs and was most abundant in the leaf (3236 µg/g extract) and stem (3093 µg/g extract), whereas the root was markedly enriched in verbascoside (2775 µg/g extract) and chlorogenic acid (1461 µg/g extract) and exhibited the highest total phenolic content, reaching 160.87 mg gallic acid equivalents (GAE)/g extract. The assay-based RACI ranked the root highest (0.54), followed by the stem (0.02), leaf (−0.27), and flower (−0.30). The root also showed the strongest 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity (half-maximal inhibitory concentration, IC₅₀ = 0.28 mg/mL) and cupric reducing antioxidant capacity (CUPRAC) (half-maximal effective concentration, EC₅₀ = 0.22 mg/mL) among the investigated organ extracts. Under the present assay conditions, all organ extracts yielded lower α-glucosidase IC₅₀ values (0.97–1.10 mg/mL) than acarbose (1.16 mg/mL). Exploratory Pearson correlation analysis indicated strong organ-level covariation among several antioxidant parameters, phenolic constituents, and α-glucosidase inhibitory activity. PCA based on the Pearson correlation matrix showed that principal components 1 and 2 (PC1 and PC2) accounted for 50.73% and 38.49% of the total variance, respectively (cumulative variance: 89.22%), providing an exploratory visualization of the multivariate organization of phytochemical and bioactivity variables. Conclusions: O. bilgeri exhibited distinct organ-specific phytochemical and bioactivity patterns under the conditions investigated. The root was particularly characterized by high verbascoside and chlorogenic acid contents, elevated total phenolic content, strong antioxidant performance across several assays, and α-glucosidase inhibitory activity. These findings support further investigation of O. bilgeri, particularly its root, using independent biological replicates and complementary phytochemical and biological approaches. Full article

ENZ-124, the first approved cetuximab biosimilar by Indian health authorities, was developed using the Chinese hamster ovary cell line, whereas the SP2/0 cell line was used for developing cetuximab originator (Erbitux^®). Hence, this assessment aimed to evaluate the physiochemical, structural, and biological characterization of four batches each of Cetuxa^®, Lupitux^® and cetuximab originator. Analytical methods like peptide mapping (liquid chromatography–tandem mass spectrometry [LC-MS/MS]), glycan mapping (liquid chromatography–fluorescence detection), isoform analysis (hydrophobic interaction chromatography [HIC]-high-performance liquid chromatography), charge variant analysis (capillary electrophoresis coupled with mass spectrometry), FcγRIIIa binding assays, cell-based antibody-dependent cell-mediated cytotoxicity (ADCC) measurement, and host cell protein analysis (LC-MS/MS) were utilized for analysis. Variations in physicochemical and biological attributes were observed between cetuximab originator and its biosimilars. HIC analysis revealed C-terminal heterogeneity and increased hydrophobicity in biosimilars, that with the different N-glycan profile affected charge distribution. Charge variant analysis indicated a more acidic profile for the biosimilars. The biosimilars showed higher oxidation levels and misincorporation. The biosimilars also showed an enhanced FcγRIIIa binding affinity and in vitro ADCC activation compared with cetuximab originator. The study outcomes highlight the ability to differentiate cetuximab originator from its biosimilars Cetuxa^® and Lupitux^® based on the observed physicochemical and biological differences. Full article

Background/Objectives: Circulating amino acids are not only markers of nutritional status; in experimental and interventional models they have been linked to hepatic lipogenesis, lipoprotein assembly, mitochondrial fatty acid oxidation, and bile acid conjugation, which makes them plausible candidate correlates of obesity-related lipid dysregulation. Despite this, most metabolomic studies of excess adiposity have focused either on a single lipid parameter—typically triglycerides—or only on branched-chain amino acids (BCAAs). This pilot study was designed to generate hypotheses about these associations across the full standard lipid panel using a targeted 19-amino acid liquid chromatography–tandem mass spectrometry (LC-MS/MS) panel in adults spanning the body mass index (BMI) spectrum, with an analytical framework oriented around lipid phenotype variance rather than body weight classification. The design is cross-sectional and the analysis exploratory; no causal or predictive claim is made. Methods: Targeted LC-MS/MS quantification of 19 plasma amino acids was performed in 50 adults grouped as normal weight (n = 20; BMI 22 ± 1.5 kg/m²), overweight (n = 20; BMI 28 ± 1.5 kg/m²), or obese (n = 10; BMI 34 ± 2.0 kg/m²). The analytical framework included: (i) one-way analysis of variance (ANOVA) with Bonferroni correction; (ii) Pearson correlation analysis; (iii) principal component analysis (PCA) performed on the lipid profile itself with amino acid projection vectors; (iv) K-means clustering based on lipid phenotype (K = 3); (v) Ward-linkage hierarchical clustering of the amino acid–lipid correlation matrix; (vi) Random Forest permutation importance for all four lipid outcomes; and (vii) composite lipid risk indices including atherogenic index (TG/HDL-C) and non-HDL cholesterol. Results: A distinct amino acid correlation pattern was observed for each of the four lipid fractions. Triglycerides (TG) correlated most strongly with glutamic acid (r = 0.58) and inversely with glutamine (r = −0.58). High-density lipoprotein cholesterol (HDL-C) correlated most strongly with glutamic acid (r = −0.61) and serine (r = 0.49). The strongest correlates of low-density lipoprotein cholesterol (LDL-C) were phenylalanine (r = 0.56) and leucine (r = 0.56), and those of total cholesterol (TC) were leucine (r = 0.63) and, inversely, glycine (r = −0.54). The atherogenic index (TG/HDL-C) increased 2.9-fold from normal to obese and was most strongly correlated with glutamic acid, isoleucine, and glycine. In the multivariable models the amino acid panel accounted for a modest share of the variance in TG (adjusted R² = 0.43; F(19,30) = 2.97, p = 0.004) and HDL-C (adjusted R² = 0.35; F(19,30) = 2.40, p = 0.016). For LDL-C and TC the adjusted R² values were close to zero (0.06 for both) and the overall models were not statistically significant (both p > 0.33); no interpretable amino acid signal was therefore present for these two fractions, and no predictors are reported for them. Lipid-based K-means clustering identified three lipid-phenotype clusters (Favorable, Intermediate, Adverse lipid profiles) with differences in amino acid z-scores. Conclusions: In this exploratory pilot cohort, lipid dysregulation in abnormal BMI was associated with two partially separable amino acid axes: a glutamic acid–glutamine axis (TG and partially HDL-C), and a glycine–serine putatively protective pattern opposing all atherogenic lipid parameters. These findings extend the established BCAA–insulin-resistance paradigm and suggest that targeted amino acid profiling—particularly for glutamic acid, leucine, glycine, and serine—may serve as a way of discovering candidate biomarkers for further study for dyslipidemia in individuals with abnormal BMI. Full article

Background/Objectives: Rheumatoid arthritis (RA) is a chronic autoimmune inflammatory disease, and ginger contains phenolic constituents with anti-inflammatory potential. This study evaluated the antiarthritic efficacy of ginger ethanolic extract (GE) and explored its potential mechanisms through integrated pharmacological and computational approaches. Methods: GE was prepared by 70% ethanol reflux extraction and administered to adjuvant-induced arthritis (AIA) rats, with methotrexate as a positive control. Body weight; paw swelling; arthritis index; spleen index; and serum interleukin-6 (IL-6), interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), and prostaglandin E2 (PGE2) were measured. Chemical profiling was performed by ultra-performance liquid chromatography–quadrupole time-of-flight tandem mass spectrometry (UPLC-Q-TOF-MS/MS), followed by network pharmacology, molecular docking, and molecular dynamics (MD) simulation. Results: GE attenuated paw edema, arthritis index elevation, splenic enlargement, and elevated serum inflammatory mediators in AIA rats. UPLC-Q-TOF-MS/MS annotated 38 constituents, mainly gingerols, shogaols, gingerdiones, gingerdiols, and related phenolic derivatives. Network analysis identified 217 overlapping ginger- and RA-associated targets, with TNF, IL6, and matrix metalloproteinase 9 (MMP9) prioritized among inflammatory and matrix-remodeling nodes. Docking and MD simulations supported stable predicted interactions for 8-gingerol-TNF and 6-gingerol-MMP9. Conclusions: GE showed antiarthritic activity in AIA rats. The integrated chemical, in vivo, and computational data suggest that ginger phenolics may modulate inflammatory mediators and candidate RA-related pathways; however, these mechanisms remain exploratory and require histological, tissue-level, and molecular validation. Full article

The widespread presence of microplastics (MPs) and associated plastic-derived compounds, bisphenols (BPs), in the environment—originating from plastic production or adsorption of environmental pollutants—results in unavoidable human exposure. Therefore, assessing their occurrence in human biological samples is essential for understanding potential health risks. In this study, pyrolysis gas chromatography–mass spectrometry was used to analyse human intraocular fluids—aqueous humour (AH) and vitreous humour (VH)—to identify 11 microplastic polymer clusters. A method based on dansyl chloride derivatisation combined with ultra-performance liquid chromatography–tandem mass spectrometry was employed to screen for five bisphenols (BPs) in blood serum collected from the same patients. Potential MPs isolated from ocular samples were qualitatively assessed by stereomicroscopy. The number of particles ranged from 14 to 69 per gram in AH and from 21 to 34 per gram in VH. Among AH samples, the most frequently detected polymer clusters were C-PVC, C-PMMA, C-PET, and C-PP, with average levels ranging from 0.41 to 0.84 µg/g. The three VH samples contained only C-PMMA, with an average concentration of 0.55 ± 0.12 µg/g. Polymers such as C-PA66 (0.30 ± 0.21 µg/g) and C-PA6 (0.15 ± 0.44 µg/g) were occasionally observed in AH. Analysis of 13 human serum samples demonstrated exposure to bisphenol A (BPA; 4.05–5.18 ng/mL) and bisphenol AF (BPAF; 2.13–4.54 ng/mL), but no clear correlation was observed between serum BP levels and intraocular MPs within the limitations of the present dataset. These findings suggest the presence of MPs and BPs in human biological samples; however, the results should be interpreted with caution owing to potential background contamination and the pilot nature of the study. Full article

Ehretia macrophylla fruit is a traditionally used but underutilized functional resource that is noted for its diverse phytochemical composition, which includes flavonoids and polyphenols. However, the systematic metabolic transformations that occur in the fruit during post-harvest storage—particularly those associated with its traditional processing into dark fruit tea—remain largely unknown, limiting efforts to optimize quality and processing strategies. In the study, to address this gap, we employed a widely targeted metabolomics approach based on ultra-performance liquid chromatography–tandem mass spectrometry (UPLC-MS/MS) to dynamically profile the fruit metabolome across four storage stages (T0, T4, T8, T12). A total of 1101 metabolites were putatively annotated, and day 8 (T8) emerged as a potential pronounced metabolic inflection point. From T8 onward, 30–40% of metabolites showed differential accumulation, characterized by a gradual rise in nutrition-relevant lipids, amino acids, and vitamins. In contrast, key antioxidant-related metabolites, including phenolics and flavonoids, increased transiently before T8 but declined sharply thereafter. These coordinated shifts correlated with an increased representation of lipid- and alkaloid-related metabolic pathways, whereas flavonoid-associated features showed a relative decline. Collectively, these findings provide insights into the biochemical processes potentially associated with fruit blackening and the marked decline in antioxidant capacity during traditional processing. Our study provides the first metabolic blueprint connecting traditional processing practices with phased metabolic remodeling, offering a scientific foundation for quality assessment and the development of informed post-harvest strategies for processing E. macrophylla fruit. Full article