Aromatic amino acids (AAAs), tryptophan, phenylalanine, and tyrosine along with their pathway metabolites have been implicated in the pathogenesis of diseases ranging from cardiovascular, neurological, inflammatory, and cancer diseases, among others. As such, the measurement of the primary AAAs, their host pathway metabolites, and microbiome derived co-metabolites in blood can provide a sensitive reflection of systemic health. The aim of the study was to develop a method for the quantification of 17 metabolites, the three AAAs and various of their metabolites in plasma using a high-throughput ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method. The method demonstrated a dynamic range (1 to 16,700 ng/mL), with detection limits (LOD) as low as 0.05 ng/mL. Quantification limits ranged from 3 to 5019 ng/mL (LLOQ) and up to 16,700 ng/mL (ULOQ). Recovery at LQC, MQC, and HQC was satisfactory and consistent across most metabolites, with significant matrix effects observed only for 4-ethylphenol sulfate. Furthermore, intra and inter-day accuracy and precision met all acceptance criteria at all quality control concentrations for most of the metabolites. Measurement of NIST SRM 1950 showcased the method's accuracy for most of the metabolites. Finally, the method was applied on the analysis of plasma samples from 55 individuals (13 males and 42 females) providing information on AAAs and their pathway metabolites relevant concentrations in human plasma.
Infertility is a complex and sensitive phenomenon experienced by many couples worldwide. Our study investigated the causal effects of 486 serum metabolites on infertility using a Mendelian randomization (MR) approach, providing insights into the etiology of infertility and available treatments. Genetic data for these serum metabolites were obtained from the Metabolomics genome-wide association study server. Infertility data were obtained from the Finnish Genome Project Consortium, version R9. Three different methods were used to explore causal effects in the MR analysis. Heterogeneity was tested using Cochran Q test. Gene fold accumulation between single nucleotide polymorphisms and outcomes was explored using MR-Egger regression and MR-Pleiotropy RESidual Sum and Outlier. We obtained a total of 19 serum metabolites causally associated with male infertility, of which 9 were protective factors and 10 were risk factors. Three serum metabolites consistently showed a strong causal association with male infertility in all MR methods. We also obtained 29 serum metabolites causally associated with female infertility, of which 15 were protective factors and 14 were risk factors. Among all complementary MR methods, 5 serum metabolites showed a strong causal association with female infertility. Cochran Q test showed no heterogeneity. MR-Egger intercept test and MR-Pleiotropy RESidual Sum and Outlier global test showed no cross-sectional pleiotropy. Of the 486 serum metabolites, a total of 19 were found to be causally associated with male infertility. Of these, 1 was identified as a risk factor and 2 as protective factors. 29 metabolites were causally associated with female infertility. Of these, 1 was identified as a risk factor and 4 as protective factors.
Evidence for causal associations between functional dyspepsia (FD) and human metabolites remains limited. Two-sample and multivariable Mendelian randomizations to detect the causal relationships between FD and human metabolites. Single nucleotide polymorphisms significantly associated with human metabolites were selected as instrumental variables, the inverse variance weighting method was used as the primary analysis method, and the results were tested for heterogeneity and horizontal pleiotropy. Genetically predicted levels of 26 circulating metabolites and 5 cerebrospinal fluid metabolites were associated with FD. Among them, Total cholesterol in large low-density lipoprotein (odds ratio [OR] = 0.985, 95% confidence interval [CI] 0.975-0.995, P = .003) and cholesteryl esters to total lipids ratio in intermediate-density lipoprotein (OR = 0.983, 95% CI 0.971-0.996, P = .008) were negatively associated with FD. It was also positively associated with the level of phenylalanine (OR = 1.037, 95% CI 1.009-1.065, P = .009) and phospholipids to total lipids ratio in small high-density lipoprotein (OR = 1.015, 95% CI 1.003-1.027, P = .017). After multivariable adjustment, FD was causally associated with phospholipids to total lipids ratio in small high-density lipoprotein (OR = 1.332, 95% CI 1.060-1.674, P = .014). Sensitivity analyses showed no evidence of heterogeneity and horizontal pleiotropy in the above results. Human metabolites may be associated with FD.
This study aimed to investigate whether interleukin-6 (IL-6) receptor inhibitors affect cerebrospinal fluid (CSF) metabolites in type 2 diabetes (T2D) patients. The initial phase employed drug-targeted Mendelian randomization to investigate the association between IL-6 receptor inhibitors and T2D across diverse populations. Subsequently, variables related to T2D were utilized as mediators in the relationship between IL-6 receptor inhibitors and CSF metabolites. Finally, the mediating effect of T2D was evaluated using mediation analysis. In the European cohort, the use of IL-6 receptor inhibitors was associated with a 25.3% reduction in the risk of T2D, as indicated by an odds ratio of 0.747 with a 95% confidence interval ranging from 0.556 to 0.938 (P = .003). In the East Asian cohort, IL-6 receptor inhibitors were associated with a 1.188-fold increase in the risk of elevated fasting insulin levels, with an odds ratio of 1.188 and a 95% confidence interval of 1.034 to 1.343 (P = .029). Mediation analysis revealed that IL-6 receptor inhibitors significantly elevated the levels of galacto-glycero-lipid (GG) in CSF, with 3.06% of the effect attributable to the reduced risk of T2D, and 96.94% directly resulting from the action of the IL-6 receptor inhibitors. The findings preliminarily indicated that race constituted a variable potentially influencing the differential effects of IL-6 receptor inhibitors among patients with type 2 diabetes (T2D) across various regions. While the administration of IL-6 receptor inhibitors might impact glucose concentrations in CSF, the role of T2D as a mediator in CSF metabolites was weak. The predominant determinant appeared to be the administration of IL-6 receptor inhibitors.
The Apocynaceae family, commonly known as the dogbane family, is widely distributed in tropical and subtropical regions and is renowned for its rich reservoir of bioactive phytochemicals. Although its plants have been extensively investigated, the fungal endophytes associated with this family remain comparatively less explored. This review provides a comprehensive synthesis of the diversity, bioactive metabolites, therapeutic potential, plant growth-promoting attributes, and nanoparticle synthesis associated with endophytic fungi of plants of Apocynaceae family. The review summarizes endophytic fungi reported from major medicinal hosts, including Catharanthus roseus, Calotropis procera, Nerium oleander, and Rauvolfia serpentina. Species belonging to the genera Alternaria, Aspergillus, Penicillium, Fusarium, Curvularia, and Cladosporium were the most frequently reported fungal endophytes. These fungi produce a wide range of secondary metabolites, including vinblastine, vincristine, taxol, oosporein, polyketides, coumarins, terpenoids, alkaloids, phenolics, and steroids, exhibiting diverse biological activities such as cytotoxic, antimicrobial, antioxidant, antidiabetic, anti-inflammatory, and insecticidal. The review further discusses emerging advances in omics-based approaches, biosynthetic gene clusters, plant growth-promoting attributes, and nanoparticle synthesis using fungal extracts, while highlighting current challenges in validating host-associated metabolites and identifying future research priorities for the sustainable exploitation of plants of Apocynaceae-associated fungal endophytes.
Neuromyelitis optica spectrum disorder (NMOSD) is a rare multifocal inflammatory disease that primarily affects the optic nerve and spinal cord, leading to visual loss and paralysis. Although current research has identified numerous biomarker changes in the cerebrospinal fluid (CSF), the causal relationships between these changes and NMOSD are not yet fully understood, and they are susceptible to confounding factors. In this study, Mendelian randomization (MR) analyses were conducted to estimate the role of CSF metabolites in aquaporin-4 (AQP4)-IgG positive and AQP4-IgG negative NMOSD. We strictly adhered to the 3 principles of MR when selecting instrumental variables, effectively reducing the impact of confounding variables. Additionally, we conducted a comprehensive and meticulous sensitivity analysis to ensure the robustness of positive outcomes. We have elucidated the roles of several CSF metabolites in NMOSD, with specific glycerophosphocholine species showing risk associations in AQP4-IgG positive and AQP4-IgG negative subtypes, while ascorbate and N-acetylhexosamines demonstrate protective effects across both subtypes, thereby providing valuable insights. No evidence of heterogeneity or horizontal pleiotropy was observed in MR analyses. These metabolites hold the potential to inform pathogenesis and guide the development of therapeutic strategies for these conditions.
Cholesterol and its metabolites are fundamental to membrane organization and cellular signaling, but their poor ionization efficiency and structural similarity complicate mass-spectrometric (MS) analysis. In this work, we developed pyridyl ethyl diazoacetate (PED) as a stable, efficient derivatization reagent that introduces a permanent positive charge on hydroxyl-containing sterols, thereby enhancing ionization and detection sensitivity. Systematic MS2 characterization established diagnostic fragmentation motifs of PED-tagged sterols, which we encoded into a query-based spectral-mining strategy for reproducible, rule-based annotation of sterol features in complex matrices. Application of the PED workflow to human liver tissues enabled sterol profiling across cirrhotic, hepatocellular carcinoma, and intrahepatic cholangiocarcinoma specimens, suggesting disease-context-dependent differences in sterol profiles. The workflow effectively integrates chemical derivatization and programmable data interrogation, providing a robust analytical framework for sterol profiling in biological samples and a potential basis for broader lipidomic applications.
Uvaria micrantha remains insufficiently characterized in terms of its phytochemical diversity and pharmacological potential. In this study, branches and leaves were extracted and subjected to chromatographic isolation, yielding a new metabolite, uvarisinyicone, along with twenty-two known compounds, including chalcones, dihydrochalcones, flavonoids, and benzenoids. LC-MS/MS profiling combined with GNPS-FBMN enabled dereplication and visualization of metabolite clusters, revealing chemical networks consistent with isolated scaffolds. Bioassays demonstrated two major functional classes of metabolites, exhibiting anti-inflammatory and anti-neuroinflammatory activity, and regulators of osteogenesis. Differential bioactivities were observed, with certain compounds selectively suppressing IL-6, others exerting dual cytokine inhibition, and several promoting osteoblast differentiations. The discovery of uvarisinyicone, together with bioactive flavonoid and chalcone derivatives, expands the chemical diversity of the genus and identifies osteogenesis activity as a novel property. These findings suggest therapeutic potential for osteoporosis and inflammatory disorders while bridging natural product chemistry with modern pharmacological applications.
Organophosphate flame retardants (OPFRs) are emerging contaminants increasingly detected in marine environments due to their extensive industrial use and replacement of brominated flame retardants. Their occurrence in different environmental compartments, combined with the formation of transformation products, raises concerns regarding persistence, transport, and ecological risks. However, the simultaneous determination of OPFRs and their metabolites in complex marine matrices remains analytically challenging due to their diverse physicochemical properties and low environmental concentrations. A sensitive and selective liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated for the simultaneous determination of eight OPFRs and three transformation products in seawater, sediments, and marine algae. The method combines solid-phase extraction for aqueous samples (200 mL) with ultrasound-assisted extraction for solid matrices, followed by chromatographic separation on a C18 column and multiple reaction monitoring (MRM) detection using electrospray ionization. Optimization of chromatographic and ionization parameters enabled the reliable separation and detection of compounds with diverse physicochemical characteristics within a single analytical run. The method demonstrated excellent linearity (R2 ≥ 0.99), satisfactory recoveries (74-105%), and good precision (relative standard deviation ≤ 10%) for all target analytes. Matrix effects, assessed by post-extraction addition, ranged from 77% to 108%, indicating only moderate signal suppression or enhancement depending on matrix type. Method detection limits ranged from 0.19 to 0.40 ng/L in seawater and remained below 0.5 ng/g in sediments and algae, confirming the suitability of the method for ultra-trace analysis. The proposed analytical approach enables the simultaneous assessment of OPFRs and transformation products in both abiotic and biotic marine compartments using a single harmonized workflow. The inclusion of algae alongside seawater and sediments provides a more comprehensive understanding of contaminant distribution and potential bioaccumulation pathways in marine ecosystems. The method represents a robust, sensitive, and reliable analytical tool suitable for environmental assessment, contamination surveillance, and large-scale marine monitoring programs.
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Metabolites produced through host-microbe interactions may help associate metabolomic signatures with therapy response. We aimed to assess serum metabolites as predictors of vedolizumab remission in children with IBD. VedoKids was a multicenter, prospective cohort study of children initiating vedolizumab at any stage of disease. Serum metabolomic profiling was performed at baseline (W0), week 14 (W14), and week 30 (W30) using quantitative metabolomics based on DI/LC-MS/MS method. Random Forest (RF) models with maximum relevance-minimum redundancy (mRMR) feature selection, integrated clinical and metabolomic features to predict clinical remission at the next time point (ie, week 0 to week 14; week 14 to week 30; week 30 to week 54). One hundred thirty-eight patients were included (mean age 13.6 years), 64 with Crohn disease (CD) and 74 with ulcerative colitis (UC). In CD, models achieved an area under the receiver operating characteristic curve (AUROC) of 0.80 (95% CI, 0.68-0.92) for predicting response to vedolizumab at W14, 0.92 (95% CI, 0.85-0.99) at W30 , and 0.83 (95% CI, 0.72-0.94) at W54. Metabolites appeared among the most important features, notably PC ae C36:4, citric acid, and malic acid. In UC, corresponding AUROC were 0.72 (95% CI, 0.59-0.85) at W14, 0.77 (95% CI, 0.64-0.90) at W30, and 0.90 (95% CI, 0.80-1.00) at W54. LysoPC a C26:1, picolinic acid, and serotonin were among the most important metabolites for prediction. We identified serum metabolites associated with vedolizumab remission in children with IBD, several of which have been previously linked to IBD pathophysiology.
Climate change has increased the incidence of compound stresses, including the co-occurrence of nitrogen deficiency (-N) and high temperature (HT), which severely reduce plant productivity. Studies have primarily focused on single plant tissues to decipher tolerance mechanisms; however, tissue-specific metabolic reprogramming remains poorly examined. This work aimed to examine the distinct metabolic reprogramming in roots and leaves under whole-plant nitrogen deficiency (-N) and high temperature (HT), applied individually or combined. We hypothesized that roots and leaves exhibit complementary metabolic profiles, while combined stress triggers a unique metabolic signature associated with plant growth regulation. Soybean plants were subjected to control, -N, HT, and HT-N conditions, and later whole-plant physiological assessment and untargeted metabolites profiling of roots and leaves were performed and analyzed by machine learning analyses (e.g., t-SNE, UMAP, WGCNA, and random forest regression), qPCR and absolute quantification of identified key metabolites. Combined HT-N stress caused severe growth inhibition, reduced shoot length (67%), root fresh weight (52%), and photosynthetic efficiency (Fv/Fm; by 51%) compared to control. Metabolomic analysis revealed stress specific responses in different tissues, with roots prioritizing N assimilation (accumulating glutamate, proline and aspartate) under -N, while leaves enhanced osmo-protection (accumulating flavonoids) under HT. Under combined HT-N, tissue-specific responses were additive, with roots focusing on amino acid and proline metabolism and leaves on phenylpropanoid and glutathione metabolism. Our machine learning analyses (t-SNE, UMAP), WGCNA and RFR showed distinct tissue-specific metabolic signatures for each stress, and identified glucose, flavonoids, proline, and specific amino acids among key candidate metabolites associated with physiological resilience. Later, exogenous application of proline, quercetin, and L-arginine recovered soybean growth under stress, but in a stress-specific manner. Soybean employs distinct metabolic strategies in roots and leaves to manage multiple stresses. The identified key metabolites represent candidate hubs in the stress response network, offering candidate targets that warrant further investigation for breeding climate-resilient crops.
Gestational diabetes mellitus (GDM) is associated with adverse pregnancy outcomes and long-term metabolic and cardiovascular risk. However, oral glucose tolerance testing at 24-28 gestational weeks limits early risk stratification. Gut microbiota-associated metabolites may reflect early metabolic abnormalities, including those relevant to cardiometabolic health, but robust early-pregnancy biomarkers remain limited. We conducted a multicenter nested case-control and prospective study involving 2,693 pregnant women. Untargeted metabolomics and metagenomics were integrated to identify GDM-associated metabolites and gut microbial alterations. Three consistently dysregulated metabolites, 3-hydroxydecanoic acid, γ-Glu-Leu, and propionic acid, were quantified by targeted LC-MS/MS. Candidate algorithms were compared using repeated 10-fold cross-validation, and a final generalized linear model was externally and prospectively validated. Women who later developed GDM showed an adverse early-pregnancy metabolic profile, including higher BMI, triglycerides, and platelet count. Untargeted metabolomics identified 14 persistently altered metabolites enriched in energy, oxidative stress, and amino acid metabolism pathways. Metagenomics revealed taxonomic restructuring and coordinated microbiota-metabolite associations. The three-metabolite model achieved AUCs of 0.838 (95% CI, 0.791-0.885) in training, 0.840 (95% CI, 0.769-0.911) in internal validation, 0.955 (95% CI, 0.925-0.985) and 0.917 (95% CI, 0.875-0.958) in two external cohorts, and 0.969 (95% CI, 0.937-1.000) in the prospective cohort. Early microbiota-associated metabolic dysregulation is detectable before routine GDM diagnosis. This compact three-metabolite panel may support early GDM risk stratification and provides metabolic evidence relevant to broader cardiometabolic risk assessment in pregnancy.
The mandarin fish, as an obligate piscivore, is highly dependent on live bait, which restricts its intensive aquaculture. Although domestication has enabled it to partially accept formulated diets, the tissue-specific molecular adaptation mechanisms of its digestive tract to artificial feed remain unclear. In this study, we conducted an integrated analysis of mandarin fish fed with live bait or artificial diet for three weeks, combining growth performance evaluation, gastric histology, and paired transcriptomic and metabolomic analyses of the stomach and pyloric ceca. AD feeding significantly improved growth performance, while histological examination revealed marked hyperplasia of the gastric mucosa and disorganized fold structures. Transcriptomic analysis identified 5065 and 3381 differentially expressed genes in the stomach and pyloric ceca, respectively. In the stomach, the artificial diet induced a glutathione-dependent antioxidant response, accompanied by glycolytic reprogramming and coordinated upregulation of genes in the extracellular matrix (ECM)-receptor interaction signaling pathway, including those encoding collagen, laminin, and integrin. In the pyloric ceca, the tricarboxylic acid (TCA) cycle and oxidative phosphorylation were broadly suppressed, whereas glycosaminoglycan degradation and lysosomal pathways were activated. Metabolomic analysis showed that gastric metabolites were enriched in vascular and inflammatory mediator pathways, while metabolites in the pyloric ceca were enriched in peroxisome proliferator-activated receptor (PPAR) signaling, sphingolipid signaling, and steroid hormone biosynthesis pathways. Following artificial diet feeding, integrated multi-omics analysis of the stomach revealed significant enrichment of pathways such as phospholipase D signaling, sphingolipid signaling, and arachidonic acid metabolism, accompanied by the accumulation of key metabolites including sphingosine-1-phosphate, 20-hydroxyeicosatetraenoic acid, and cellobiose. Integrated analysis of the pyloric ceca identified significantly altered pathways, including sphingolipid metabolism, alpha-linolenic acid metabolism, and glutathione metabolism, along with elevated levels of sphingosine-1-phosphate, sphingosine galactoside, and 9-hydroxy-12-oxo-10,15-octadecadienoic acid, as well as decreased glutathionylspermidine. These findings systematically unveil the tissue-specific molecular adaptation characteristics of the mandarin fish digestive tract in response to artificial feed, providing an important basis for understanding the molecular mechanisms of dietary adaptation in carnivorous fish and for optimizing artificial feed formulations.
The pathophysiology of acute pancreatitis (AP), a common clinical emergency, is poorly understood. Previous studies have implicated N6-methyladenosine (m6A) modification in the pathogenesis of AP; however, the precise molecular mechanisms remain unclear. Whether metabolites participate in this process is also an important, unresolved question. To investigate the possible causal pathways, we integrated multi-omics data. We identified m6A related genes from literature and combined these genes with genetic summary statistics of AP, expression quantitative trait loci (eQTL) data, and information on 1400 plasma metabolites from public databases. We constructed a causal inference framework based on these data. The causal relationship between m6A-related genes, metabolites, and AP risk was systematically evaluated using Mendelian randomization and mediating Mendelian randomization methods. To validate the reliability of our results, we performed leave-one-out sensitivity, heterogeneity tests, and horizontal pleiotropy. We found that the fat mass and obesity (FTO) eQTL was significantly negatively correlated with AP risk (odds ratio [OR] = 0.78, 95% confidence interval [CI]: 0.66-0.93, P < .01). In addition, we found that the FTO eQTL reduced the level of 1,2-dilinoleoyl-glycerophosphocholine (1,2-dilinoleoyl-GPC, 18:2/18:2; OR = 0.89, 95% CI: 0.80-0.99, P < .05) and that a high level of this metabolite increased the risk of AP (OR = 1.22, 95% CI: 1.08-1.39, P < .01). Therefore, we constructed and verified a regulatory axis named "FTO-m6A-1,2-dilinoleoyl-GPC (18:2/18:2) ": Through its role in diminishing m6A modification levels, FTO-mediated demethylation lowers circulating 1,2-dilinoleoyl-GPC (18:2/18:2), which contributes to the suppression of AP. This is the first time that we have discovered the "FTO-m6A-1,2-dilinoleoyl-GPC (18:2/18:2)" regulatory axis in AP. We found that FTO may promotes phospholipid metabolic reprogramming by stimulating an m6A-dependent posttranscriptional mechanism, which further inhibited disease development. These findings provide new insights into the process of AP and new directions for targeted therapy.
Prenatal exposure to metal mixtures may influence newborn birth size, but the underlying metabolic mechanisms remain unclear. This study aimed to evaluate the associations of 19 individual maternal metals and metal mixtures during pregnancy with birth size indicators, specifically birth weight z score (BWZ) and birth length z score (BLZ), and to identify potential metabolic pathways and mediating metabolites based on cord blood metabolomics. A total of 1059 mother-infant pairs were included in the full cohort. We applied ExWAS, ENET/DSA, WQS, qgcomp, and BKMR to assess both single-metal and joint exposure effects. Exploratory MWAS, pathway enrichment, and mediation analyses were conducted using cord blood metabolomics data. In single-metal analyses, Zn, Se, and Mn showed positive associations with BWZ and BLZ, whereas Cu was positively associated with BWZ only; As, Pb, and Cd generally showed inverse associations. In mixture analyses, the negative-direction WQS index was associated with lower BLZ (β = -0.039, P = 0.042). Exploratory metabolomics analyses highlighted lipid-related metabolites, organic acid derivatives, and nucleotide-related metabolites, with enrichment in caffeine metabolism and steroid hormone biosynthesis pathways. Mediation analyses suggested potential indirect effect signals, including As-BWZ via 10-hydroxystearic acid (19.6%), Cu-BWZ via 2-ethyl-2-hydroxybutyric acid (30.8%), and Se-BLZ via 2-ethyl-2-hydroxybutyric acid (21.1%). In this industrial city cohort, our findings provide evidence linking prenatal metal exposure to newborn birth size and suggest that cord blood metabolic profiles may offer exploratory mechanistic clues for understanding metal-related fetal growth patterns.
Pharmaceutical contamination by antibiotics poses a major environmental challenge, particularly as regulators set limits for treated wastewater effluents. This study presents a hitherto unreported application of pulsed high-frequency sonication for degrading trimethoprim (TMP) and associated antimicrobial resistance genes (ARGs) in synthetic and reject wastewater. Optimization with synthetic binary samples revealed that sonication achieved >80% TMP removal at calorimetric-based energy per order (EEO) of 116.54 kW/m3, but mineralization remained low (∼10%). Prolonged treatment (5-6 h) fully eliminated TMP, yielding heavily hydroxylated and probably substituted products with short-chain alcohol groups, which exhibited lower biodegradability than demethylated/demethoxylated metabolites (e.g. m/z 245) dominant at 1-4 h. Notably, 30 min sonication produced hydroxylated TMP (m/z 307) with slight cytotoxicity to HeLa cells. The diaminopyrimidine (DAP) moiety of TMP, which drives its antimicrobial activity, remained largely intact in most metabolites identified in all the treatment times. This indicates that extending the treatment time would not provide additional benefits. Rather, sonication-based degradation should be tuned to promote site-specific attacks on antibiotic molecules, thereby inhibiting antimicrobial activity and minimizing the risk of ARGs development in treated water. Combining high-frequency sonication with low dose ozonation (via Venturi injection) effectively removed TMP (>98%) from low-solids reject wastewater, alongside substantial reductions in solids and COD. However, at higher solids levels, the process may induce cell lysis and release of TMP and ARGs from particulates. These findings position high-frequency ultrasound assisted ozonation as an effective treatment for wastewater streams with low solids, though careful control is needed to mitigate ARG mobilization in turbid matrices.
Tuberculous meningitis (TBM), caused by Mycobacterium tuberculosis, is the most severe form of extrapulmonary tuberculosis and isassociated with high morbidity and mortality, particularly when diagnosis is delayed. Improved understanding of the metabolic alterations associated withTBM may support the development of novel diagnostic biomarkers and provide insights into disease pathophysiology. In this study, we applied anuntargeted two-dimensional gas chromatography-time-of-flight mass spectrometry (GC×GC-TOFMS) metabolomics approach to formalin-fixed, paraffin-embedded (FFPE) postmortem human brain tissue from 41 TBM cases and 36 tissue sections from 6 non-TBM control cases. Metabolomics data wereprocessed, normalized, and analyzed using multivariate and univariate statistical approaches, including principal component analysis (PCA) and partialleast squares-discriminant analysis (PLS-DA), with variable importance in projection (VIP) scores. These results were further correlated with patient clinical data. Distinct metabolic profiles were observed between TBM and control tissues. Several metabolites were significantly reduced in TBM samples, particularly within the alkane and alkene classes, with additional decreases observed in metabolites associated with alcohols, fatty acids, lipids, carbohydrates, and amino acids. These metabolic alterations suggest substantial perturbations, primarily in the host lysine degradation pathway (linked to the kynurenine pathway), in TBM-affected brain tissue. Collectively, these findings provide insight into the metabolic landscape of terminalTBM and suggest potential metabolic pathways that may contribute to disease pathophysiology. Further investigation of these metabolic signatures in accessible patient tissue and biofluids may support the development of biomarkers and inform future therapeutic strategies for TBM.
Liupao tea (LPT) undergoes significant bioactive changes during aging, while the link between these chemical shifts and the regulation of oxidative stress pathologies remains unclear. Therefore, this study integrated metabolomics, network pharmacology, and molecular docking to analyze Maosheng (MS) and Tianyu (TY) LPT samples aged 0-15 and 0-10 years, respectively, and elucidated the potential mechanisms of LPT in five oxidative stress - related diseases, including depression, obesity, Alzheimer's disease, diabetes, and hypertension. Metabolomics identified 42 and 13 core antioxidant metabolites in MS and TY samples, respectively. Among them, core metabolites in MS peaked at 5 years, while TY sustained high abundance levels from 3 to 8 years. Network pharmacology revealed synergistic regulation of all five pathologies by both LPTs, and eight key components were screened from MS, and nine from TY based on degree values. Interestingly, the highest node density was shown in depression, and Sankey connectivity confirmed its strongest modulatory effect. Besides, the core depression‑related components and targets were subjected to molecular docking simulations, and binding energies ranged from - 5.37 to -8.88 kcal/mol for MS components, as well as from - 4.11 to -8.39 kcal/mol for TY components. Importantly, oleaside A and CCRIS 7793 showed the strongest affinities for GAPDH and AKT1, respectively. In general, these results laid a practical foundation for developing an LPT product aimed at relieving oxidative stress-related health issues.
Magnetic resonance spectroscopy (MRS) is a complementary imaging technique enabling non-invasive metabolic characterization of tissues in clinical practice. Proton MRS is more common for analysis of metabolites such as lactate and choline. This is valuable for preoperative assessment of glioma grade. However, it does not directly reflect phosphate energy metabolism. Phosphorus MRS enables assessment of phosphorus-containing compounds involved in tumor energy metabolism and membrane turnover. To assess the role of phosphorus MRS in metabolic characterization of low-grade and high-grade gliomas. The study included 46 patients with gliomas: 31 patients with high-grade gliomas and 15 patients with low-grade gliomas. The control group comprised 15 healthy volunteers. All participants underwent phosphorus MRS with assessment of spectral parameters of phosphorus-containing metabolites, metabolic ratios and intracellular pH. Phosphorus MRS revealed differences in metabolic profile of tumor tissue between high-grade and low-grade gliomas. High-grade gliomas were characterized by signs of impaired energy and membrane metabolism, including changes in phosphorus-containing metabolite ratios and shift in intracellular pH. Low-grade gliomas demonstrated less severe metabolic alterations. Phosphorus MRS is a promising complementary imaging modality for non-invasive assessment of metabolic features of gliomas. This method may be used to refine their biological characteristics at the preoperative stage. Магнитно-резонансная спектроскопия (МРС) является дополнительной и уточняющей методикой, позволяющей неинвазивно изучать метаболические свойства тканей. В клинической практике чаще используется протонная МРС, позволяющая анализировать такие метаболиты, как лактат и холин, что помогает в дооперационной характеристике глиом. Однако она не отражает напрямую обмена энергетических фосфатов. Фосфорная МРС позволяет оценивать фосфорсодержащие соединения, связанные с энергетическим и мембранным метаболизмом опухоли. Оценить возможности фосфорной МРС в характеристике метаболического профиля глиом различной степени злокачественности. В исследование включены 46 пациентов с глиомами: 31 пациент с глиомами высокой степени злокачественности и 15 — с глиомами низкой степени злокачественности. Контрольную группу составили 15 здоровых добровольцев. Всем участникам была выполнена фосфорная МРС с оценкой спектральных показателей фосфорсодержащих метаболитов, а также расчетных метаболических соотношений и внутриклеточного pH. Фосфорная МРС позволила выявить различия метаболического профиля опухолевой ткани при глиомах высокой и низкой степени злокачественности. Для высокозлокачественных глиом были характерны признаки нарушения энергетического обмена и мембранного метаболизма, включая изменение соотношений фосфорсодержащих метаболитов и сдвиг внутриклеточного pH. Низкозлокачественные глиомы демонстрировали менее выраженные метаболические изменения. Фосфорная МРС является перспективной дополнительной методикой неинвазивной оценки метаболических особенностей глиом и может использоваться для уточнения их биологических характеристик на дооперационном этапе.