Plasma Metabolomics Research Articles

Development and Validation of Targeted Metabolomics Methods Using Liquid Chromatography–Tandem Mass Spectrometry (LC-MS/MS) for the Quantification of 235 Plasma Metabolites

Plasma contains metabolites with diverse physicochemical properties, ranging from highly polar to highly apolar, and concentrations spanning at least nine orders of magnitude. Plasma metabolome analysis is valuable for monitoring health and evaluating medical interventions but is challenging due to the metabolome’s diversity and complexity. This study aims to develop and validate targeted LC-MS/MS methods for quantifying 235 mammalian metabolites from 17 compound classes in porcine plasma without prior derivatization. Utilizing reversed-phase and hydrophilic interaction liquid chromatography coupled with tandem mass spectrometry, each analyte is identified and quantified using two selected reaction monitoring (SRM) transitions. Fast polarity switching and scheduled SRM enhance the metabolome coverage and throughput, enabling the analysis of one sample in about 40 min. A simple “dilute and shoot” sample preparation protocol was employed, with samples injected at two dilution levels to align metabolite concentrations within calibration curve ranges. Validation in porcine plasma included assessments of carryover, linearity, detection and quantification limits, repeatability and recovery. The method was further applied to plasma samples from various animal species, demonstrating its applicability to human and animal studies. This study establishes two robust LC-MS/MS methods for comprehensive porcine plasma metabolome quantification, advancing large-scale targeted metabolomics in biomedical research.

The Metabolic Profile of Plasma During Epileptogenesis in a Rat Model of Lithium-Pilocarpine-Induced Temporal Lobe Epilepsy.

Temporal lobe epilepsy (TLE) arises mostly because of an initial injury. Certain stimuli can make a normal brain prone to repeated, spontaneous seizures via a process called epileptogenesis. This study examined the plasma metabolomics profile in rats with the induced TLE to identify feasible biomarkers that can distinguish progression of epileptogenesis in three different time points and reveal the underlying mechanisms of epileptogenesis. Status epilepticus (SE) was induced by repetitive intraperitoneal injections of low-dose lithium chloride-pilocarpine hydrocholoride. Blood samples were collected 48h, 1week, and 6weeks after SE, respectively. Plasma metabolites were analyzed by nuclear magnetic resonance (NMR) spectrometry. Statistical analysis was performed using MetaboAnalyst 6.0. An orthogonal partial least squares discriminant analysis (OPLS-DA) model was employed to represent variations between the TLE model groups and respective controls. Volcano plot analysis was used to identify key features, applying a fold-change criterion of 1.5 and a t-test threshold of 0.05. 48h after SE, dimethyl sulfone (DMSO2) and creatinine levels were decreased, whereas glycine and creatine levels were increased. The only metabolite that changed 1week after SE was pyruvic acid, which was increased compared to its control level. Lactic acid, pyruvic acid, and succinic acid levels were increased 6weeks after SE. The identified metabolites were especially related to the tricarboxylic acid cycle and glycine, serine, and threonine metabolism. The results illustrate that distinct plasma metabolites can function as phase-specific biomarkers in TLE and reveal new insights into the mechanisms underlying SE.

Genetically Predicted Plasma Metabolome Mediates the Causal Link Between Immune Cells and Risk of Gout.

Gout is a prevalent metabolic disorder characterized by a multifaceted process of development. Recent research has emphasized a robust correlation between the immune response and gout. Nevertheless, it is still uncertain if this connection is causative. Hence, the objective of this study was to investigate the causal relationship between immune cells and gout, while also analyzing the role of the plasma metabolome as metabolic mediators in this biological process. This study explored the causal link between different subtypes of immune cells and gout using two-sample Mendelian randomization (MR). To confirm the reliability of the findings, reverse MR analysis, steiger test and sensitivity tests were conducted. A two-step mediation analysis was used to gain insight into the role of plasma metabolites as intermediate mediators. This two-sample, bidirectional, two-step MR analysis found a nominal causal link between 33 immune cells as well as 47 known plasma metabolites and gout. Reverse MR analysis and sensitivity tests demonstrated the reliability of the MR results. In addition, we found that Tetradecadienedioate (C14:2-DC) played a partially mediating role in the CD4 on activated CD4 regulatory T cell and gout pathways, with a mediating proportion of 13.16%, (95% CI = 0.65%-25.67%, p = 0.034). The objective of our research was to investigate the possible causative connection between immune cells and gout. Our findings indicate that certain plasma metabolites may play a role in mediating this association. This study offers novel insights and sources of information that may contribute to the early detection and proactive measures to avoid gout in the future.

Analysis of ischemic stroke biomarkers based on non-targeted metabolomics

Biomarkers for ischemic stroke (IS) are yet to fulfill clinical requirements. This study used non-targeted metabolomics to investigate differential metabolites and metabolic pathways in plasma and brain tissue following IS, with the aim of identifying new potential biomarkers and therapeutic targets. Twelve Tibetan miniature pigs were randomly assigned to a model- or sham-operation group. An electrocoagulation-based anterior temporal approach was employed to occlude the middle cerebral artery, thereby creating a model for IS. Plasma and brain tissue samples were collected 36 h post-surgery and analyzed using liquid chromatography-mass spectrometry. Principal component and partial least squares discriminant analyses were used to screen for differential metabolites and exclude exogenous metabolites at p<0.05. Compounds were classified according to the HMDB (Human Metabolome Database), and subjected to KEGG (Kyoto Encyclopedia of Genes and Genomes) pathway and VIP (variable importance in projection) analyses. Plasma metabolomics revealed that 86 metabolites were upregulated while 149 were downregulated, with (Z)-3-oxo-2-(2-pentenyl)-1-cyclopentylacetic acid, trans-cinnamic acid and cinnamoylglycine determined to be significant metabolites. Fifty-eight differential metabolites were upregulated in brain tissue and 53 were downregulated, with 2,3-dihydroflavon-3-ol, guanidinoacetic acid (GAA), N-acetyl-D-tryptophan, oxidized glutathione, 2-hydroxyquinoline, and N-acetyl-L-aspartate (NAA) identified as significant metabolites. Organic acids and derivatives, lipids and lipid-like molecules, organoheterocyclic compounds, and organic oxygen compounds were found to be common compound categories among the top five types of compound in both plasma and brain tissue. Common metabolic pathways in plasma and brain tissue include amino acid metabolism, digestive system, cancer overview, and lipid metabolism pathways, with the (Z)-3-oxo-2-(2-pentenyl)-1-cyclopentylacetic acid, GAA, oxidized glutathione, and NAA metabolites serving as potential biomarkers. This study provides a theoretical foundation for the early screening and development of clinical treatment strategies for IS.

Pulsed red light photobiomodulation ameliorates oxytocin-induced primary dysmenorrhea in mice by inhibiting oxidative stress and lipid accumulation.

Photobiomodulation (PBM) has gained attention as a kind of anti-pain or anti-inflammation therapy, yet its efficacy in mitigating the symptoms and underlying metabolic disturbances of primary dysmenorrhea remains underexplored. Here, 630nm light reduced menstrual pain and prostaglandin F2a/prostaglandin E2 dysregulation, regulated oxidation and lipid peroxidation levels, and improved uterus damage in oxytocin-induced mice. Notably, pulsed wave (PW) treatment exhibited superior efficacy compared to continuous wave application. Hence, this research focused on the effects of 630nm PW on oxytocin-induced mice by examining changes in the uterine transcriptome and plasma metabolome. Results from integrated analyses revealed significant modifications primarily in antioxidant and lipid metabolism pathways, alongside shifts in biomarkers related to arachidonic acid metabolism. Quantitative real-time PCR confirmed the downregulation of critical genes associated with oxidative stress and inflammation, as well as the suppression of uterine smooth muscle contractions and lipid overaccumulation. These findings support the potential of 630nm PW PBM as a viable option for clinical interventions in dysmenorrhea management.

Seminal plasma metabolomics and sperm lipidomics profiles of bull semen with different total progressive motile sperm count.

Total progressive motile sperm count (TPMSC) is a reliable index of fecundity evaluation of bull semen. It is an important determinant frozen semen yield and conception rate of females artificially inseminated. Seminal plasma metabolites and sperm lipids are closely related to sperm survival and motility, but their relationship with TPMSC is not well known. In the present study, Simmental bulls with higher (H, n = 6) or lower (L, n = 6) TPMSC (P < 0.01) were selected from a cohort of 100 animals aged 2 to 5 years based on semen quality. Analysis of semen quality and biochemical markers of seminal plasma revealed that H bulls had greater ejaculate volume (P < 0.05), sperm motility, plasma membrane integrity rate (P < 0.01), seminal plasma neutral α-glucosidase (P < 0.05), alkaline phosphatase, acid phosphatase, cortisol and phosphatidylcholine (P < 0.01), and lower sperm malformation rate (P < 0.05) and reactive oxygen species (ROS) (P < 0.01). Semen metabolites and sperm liposome profiles of H and L groups were compared using LC-MS/MS analysis. A total of 120 differentially abundant metabolites (VIP > 1; P < 0.05) and 59 differentially abundant lipids (VIP > 1; P < 0.05) were identified between H and L groups. Oxidative stress, sperm motility and sperm plasma membrane integrity were among the enriched biological pathways. Cyclic ADP-ribose (cADPR), up-regulated in H bulls, is associated with energy for sperm motility and maintenance of membrane stability. Thymidineglycol (Tg), levanbiose, thymidine (Thd), and CE (3M5) were down-regulated in H bulls, and may have negatively affected sperm motility. Correlation analyses revealed that TPMSC and sperm motility were significantly positively correlated with cADPR, while Tg, Levanbiose, Thd and CE (3M5) were significantly negatively correlated with TPMSC and sperm motility. Thus, we speculate that these molecules may be exploited as potential biomarkers for non-invasive evaluation of TPMSC in bull semen.

Multi-omics dissection of metabolic dysregulation associated with immune recovery in people living with HIV-1

BackgroundDespite the success of antiretroviral therapy (ART) in suppressing HIV-1 replication, some people living with HIV-1 (PLWH) fail to achieve an optimal recovery of CD4 T cells, and precise metabolic regulation underlying immune recovery remained poorly understood.MethodsIn this cross-sectional study, mass spectrometry was used for quantitative analysis of plasma metabolome and lipidome in 24 treatment-naïve PLWH (TNs), 33 immunological responders (IRs), 35 immunological non-responders (INRs), and 16 healthy controls (HCs). The data were analyzed using the Mann–Whitney U-test, Kruskal–Wallis test, Spearman correlation, and LASSO regression analysis.ResultsSignificant metabolic dysregulation was observed in TNs, IRs and INRs compared to HCs. In TNs, metabolomic analysis revealed increased levels of 3-hydroxyoctanoic acid, 3-oxododecanoic acid, 5-hydroxy-L-tryptophan, 5-hydroxyindoleacetic acid, L-kynurenine, oleoylcarnitine, and pseudouridine that were positively correlated with CD8 T cell activation and inflammation-related markers, and decreased levels of phosphorylcholine, ribothymidine, and thymine that were negatively correlated. Notably, 3-hydroxyoctanoic acid and thymine were consistently associated with CD4 T cell counts and inflammation-related markers in PLWH, regardless of ART. Pathway analysis uncovered the biosynthesis of unsaturated fatty acids as the major dysregulated pathway in TNs, IRs, and INRs, while primary bile acid biosynthesis was the dysregulated pathway specifically in INRs. Lipidomic analysis indicated higher plasma triacylglycerols, free fatty acids, ceramide, and monosialodihexosyl gangliosides (GM3) in TNs, IRs, and INRs compared to HCs. Pathway enrichment and differential correlation analyses underscore perturbed systemic lipid metabolism in treatment response to ART, possibly mediated by host-commensal metabolic interactions. Ultimately, we identified two panels, one consisting of 9 metabolites and another of 8 lipids, that can effectively distinguish INRs from IRs.ConclusionsMetabolic aberrations induced by chronic HIV-1 infection persist and do not recover with ART. Abnormal primary bile acid biosynthesis pathway and levels of DHA-containing lipids are closely associated with CD4 T cell recovery. These finding provide new intervention targets to achieve better immune recovery in PLWH.

Effect of SY009, a novel SGLT1 inhibitor, on the plasma metabolome and bile acids in patients with type 2 diabetes mellitus

ContextAs a novel SGLT1 inhibitor, SY-009 has been preliminarily confirmed in a phase Ib clinical study for its ability to reduce postprandial blood glucose in patients with type 2 diabetes mellitus (T2DM). However, the effects of SY-009 on human plasma metabolomics are still unknown.ObjectiveThis study aimed to explore the effects of SY-009 on plasma metabolomics in patients with T2DM and the potential metabolic regulatory mechanism involved.Study designIn the phase Ib study, a total of 50 participants with T2DM were enrolled and randomly assigned to the 0.5 mg BID, 1 mg BID, 2 mg BID, 1 mg QD, and 2 mg QD dose groups, with a 4:1 random allocation within each group to receive either the SY-009 capsule or placebo. We conducted untargeted and targeted metabolomics analyses on plasma samples from the phase Ib clinical study.ResultsUntargeted metabolomics revealed that, after SY009 treatment, there were differences in metabolic pathways, including primary bile acid biosynthesis; biosynthesis of unsaturated fatty acid; steroid hormone biosynthesis; purine metabolism; phenylalanine, tyrosine and tryptophan biosynthesis. In particular, the increase in bile acid-related metabolites in the 2 mg BID group was significantly greater than that in the placebo group, and unsaturated fatty acid-related metabolites decreased in both the 2 mg BID group and the placebo group, but there was no significant difference between the two groups. After comprehensive consideration, bile acids were taken as our target for accurate quantification via targeted metabolomics. Compared with those in the placebo group, the levels of several bile acids were significantly greater in the SY-009-treated groups. Moreover, the proportion of free bile acids decreased significantly, the proportion of glycine-conjugated bile acids increased significantly, the proportion of taurine-conjugated bile acids tended to be stable, and PBA/SBA significantly increased after SY-009 administration.ConclusionsSY-009 caused a series of postprandial plasma metabolite changes in patients with T2DM, especially significant changes in the bile acid profile, which provides a new perspective on the mechanism by which SY-009 lowers blood glucose.Clinical trial registrationhttps://www.clinicaltrials.gov, identifier NCT04345107.

Impact of variations in airborne microbiota on pneumonia infection: An exploratory study.

Previous studies showed airborne bacteria affect pneumonia incidence, but specific impacts of bacterial communities on Klebsiella pneumoniae infection were unknown. Five different ratios of bacterial community structures were randomly generated. Mice were divided into control, artificial bacterial community exposure, and corresponding Klebsiella pneumoniae challenge groups. Changes in body weight, blood parameters, pulmonary pathology, inflammatory factors, metabolomics, and fecal microbiota were analyzed. Different bacterial community exposures had varying degrees of influence on body weight, complete blood count, inflammatory factors, alveolar lavage fluid and plasma metabolome, as well as intestinal microbiota at baseline and after infection. Metabolomic analysis showed that microbial exposure affected both bronchoalveolar lavage fluid and plasma metabolomes, suggesting systemic effects of microbial exposure on the organism. Differences in the structure of artificial microbiota had inconsistent effects on both the baseline state and the post-infection state, hinting at crosstalk between microbial exposure and Klebsiella pneumoniae infection. KEGG pathway analysis unveiled possible molecular mechanisms underlying the overall impact of microbial exposure on the lungs and the body as a whole. In the intestinal microbiota, differences were found in composition at the phylum and genus levels. Spearman correlation analysis established potential correlations between intestinal microbiota and differential metabolites, suggesting a potential link within the lung-gut axis. This study demonstrated the significant and systemic impact of air microbiota structure differences on health. Future research should explore the underlying mechanisms to enhance our understanding of the air-environment-health relationship and identify interventions for improving public health strategies.

Metabolome Alterations Associated with Three-Month Sitting-Time Reduction Among Sedentary Postmenopausal Latinas with Cardiometabolic Disease Risk

Background: Incidence of cardiometabolic disease among U.S. Hispanics/Latinos is higher than in non-Hispanic Whites. Prolonged sitting duration is prevalent in older adults, and compounded with menopause, greatly increases cardiometabolic risk in postmenopausal women. Metabolomic analyses of interventions to reduce sitting are lacking and mechanistic understanding of health-promoting behavior change in postmenopausal Latinas is needed. Methods: To address this knowledge gap, an exploratory analysis investigated the plasma metabolome impact of a 12-week increased standing intervention among sedentary postmenopausal Latinas with overweight or obesity. From a parent-randomized controlled trial, a subset of Best Responders (n = 43) was selected using parameters of highest mean change in sitting bout duration and total sitting time; baseline variable-Matched Controls (n = 43) were selected using random forest modeling. Targeted LC-MS/MS analysis of archived baseline and 12-week plasma samples was conducted. Metabolite change was determined using a covariate-controlled general linear model and multivariate testing was performed. A false discovery rate correction was applied to all analyses. Results: Best Responders significantly changed time sitting (−110.0 ± 11.0 min; −21%), standing (104.6 ± 10.1 min; 40%), and sitting in bouts &gt;30 min (−102.3 ± 13.9 min; −35%) compared to Matched Controls (7.1 ± 9.8 min, −7.8 ± 9.0 min, and −4.6 ± 12.7 min, respectively; all p &lt; 0.001). Twelve-week metabolite change was significantly different between the two groups for 24 metabolites (FDR &lt; 0.05). These were primarily related to amino acid metabolism, improved blood flow, and ATP production. Enzyme enrichment analysis predicted significant changes regulating glutamate, histidine, phenylalanine, and mitochondrial short-chain fatty acid catabolism. Pathway analysis showed significant intervention effects on glutamate metabolism and phenylalanine, tyrosine, and tryptophan biosynthesis, potentially indicating reduced cardiometabolic disease risk. Conclusions: Replacing nearly two hours of daily sitting time with standing and reduced prolonged sitting bouts significantly improved metabolomic profiles associated with cardiometabolic risk among postmenopausal Latinas.

Effects of rumen-degradable starch on lactation performance, gastrointestinal fermentation, and plasma metabolomic in dairy cows.

This study investigated the effects of rumen-degradable starch (RDS) on lactation performance, gastrointestinal fermentation, and plasma metabolomics in dairy cows. Six mid-lactation cows, fitted with rumen, duodenum, and ileum cannulas, were used in a duplicated 3 × 3 Latin square design with 28-day periods. The cows were fed a low RDS (LRDS; 62.18 %), medium-RDS (MRDS; 71.25 %), or high-RDS (HRDS; 80.32 %) diet. The results showed that cows fed HRDS had diet a lower milk fat content by 14.77 % (LRDS) and 11.73 % (MRDS), while increased somatic cell count compared to the LRDS and MRDS groups (34.42 and 29.38 %, respectively). Additionally, rumen fluid pH was decreased in the HRDS group than in the MRDS and LRDS groups (7.81 and 7.08 %, respectively), while microbial protein (MCP) concentration was higher in the MRDS group. The HRDS group had lower concentrations of total volatile fatty acids (TVFA) and acetate in the ileal digesta than the LRDS group. The HRDS diet decreased neutral detergent fibre (NDF) digestibility compared with LRDS and MRDS groups (7.68 and 8.50 %, respectively), and reduced plasma concentrations of superoxide dismutase (SOD) and glutathione peroxidase (GSH-PX), while increasing plasma lipopolysaccharide (LPS) and serum amyloid-A (SAA) levels. Pathway analysis revealed that starch and sucrose metabolism, galactose metabolism, and carbohydrate digestion and absorption were upregulated in the MRDS and HRDS groups. The HRDS diet had a tendency to negatively affect linoleic acid metabolism, glycerophospholipid metabolism, and alpha-linolenic acid metabolism. These findings provide insights into optimising feed efficiency and milk quality by regulating RDS levels in dairy cow diets.

Role of plasma metabolome in mediating the effect of plasma lipidome on NAFLD: a Mendelian randomization study

Role of plasma metabolome in mediating the effect of plasma lipidome on NAFLD: a Mendelian randomization study

Perturbations in plasma amino acid and lipoprotein subfraction profiles in anorexia nervosa before and after refeeding: A metabolomic cross-sectional and longitudinal analysis.

Anorexia nervosa (AN) is a life-threatening eating disorder, which is increasingly being considered a metabo-psychiatric condition. We aimed to assess how the lipoprotein subfraction and plasma metabolome are altered in acutely underweight patients with AN (AcAN), if they change with short-term weight-restoration, and whether these changes point towards altered cardiometabolic risk. Using nuclear magnetic resonance spectroscopy, we measured and compared the plasma concentrations of 132 metabolites, aminoacids and lipoprotein subfractions in young female patients with AcAN before (n=72) versus after (n=46) a short-term inpatient refeeding program resulting in weight-restoration (longitudinal analysis), as well as versus female healthy control (HC) participants of similar age (n=74) (cross-sectional analysis). Patients with AcAN showed elevated plasma cholesterol levels due to higher concentrations of small and dense Low Density Lipoprotein (LDL-6) and of large and less dense High Density Lipoprotein (HDL-1) subfractions compared to HC. Additionally, they had lower plasma concentrations of branched chain amino acids and glucose and higher concentrations of the gluconeogenic amino acids glutamine, alanine and methionine. Refeeding elevated the plasma cholesterol levels further, but with a different pattern compared to AcAN, by increasing the concentrations of the larger and less dense LDL (LDL-1, LDL-2, LDL-3) particles and of smaller and more dense HDL (HDL-2, HDL-3) subfractions. However, refeeding only partially restored the amino acid concentrations. Lipoprotein profiles in AcAN point towards a potentially elevated risk for atherosclerosis; an altered lipoprotein profile was also detected after refeeding. Metabolite profiles in AcAN indicate an advanced catabolic state with only partial restoration after refeeding.

A new model of experimental diabetic cardiomyopathy using combination of multiple doses of anomer-equilibrated streptozotocin and high fat diet: sex matters.

Diabetes mellitus (DM) leads to a more rapid development of DM cardiomyopathy (dbCM) and progression to heart failure in women than men. Combination of high-fat diet (HFD) and freshly-injected streptozotocin (STZ) has been widely used for DM induction, however emerging data shows that anomer-equilibrated STZ produces an early onset and robust DM model. We designed a novel protocol utilising a combination of multiple doses of anomer-equilibrated STZ injections and HFD to develop a stable murine DM model featuring dbCM analogous to humans. Furthermore, we examined the impact of biological sex on the evolution of cardiometabolic dysfunction in DM. Our study included six experimental protocols (8 weeks) in male and female C57BL/6J mice (n=109): Fresh STZ+HFD, Anomer-equilibrated STZ+HFD, HFD, Fresh STZ, Anomer-equilibrated STZ, Control diet+vehicle. Animals were characterised by extensive phenotyping in vivo and ex vivo. Anomer-equilibrated STZ+HFD led to induction of stable experimental murine DM characterised by impaired glucose homeostasis, cardiometabolic dysfunction and altered metabolome of liver, skeletal muscle, kidney and plasma. dbCM was more severe in female mice including systolic dysfunction and reduced cardiac energy reserve. This study established a novel, robust model of inducible murine DM and emphasised the impact of biological sex on DM progression and severity.

Effects of patent foramen ovale in migraine: a metabolomics-based study.

Patent foramen ovale (PFO), a cardiac anatomical anomaly inducing abnormal haemodynamics, leads to a paradoxical bypass of the pulmonary circulation. PFO closure might alleviate migraines; however, clinical evidence and basic experiments for the relationship are lacking. To explore the effect of PFO on migraine, 371 migraineurs finishing blood tests and contrast transthoracic echocardiography for the detection of PFO were prospectively included. Multivariate regression analysis revealed that PFO was independently associated with aura, and lower cystatin-C (cys-C) and calcium levels. Among them, patients with PFO who underwent percutaneous PFO closure were continuously followed up 1year after the operation. The intensity of migraine was significantly relieved and the levels of cys-C and calcium increased after PFO closure. Untargeted and targeted metabolomics of plasma from migraineurs before and after PFO closure revealed that 5-HT and glutathione (GSH) metabolites were differentially expressed after PFO closure. The differential metabolites were then validated in the plasma and brain tissues of PFO mouse models by LC-MS/MS analysis. Desorption electrospray ionization mass imaging demonstrated that these metabolic alterations occurred mainly in the posterior cerebral cortex. Collectively, aura, cys-C and calcium could be biomarkers of migraineurs with PFO. PFO might have an impact on the posterior head associated with the regulation of 5-HT and GSH. PFO closure might relieve migraine by improving 5-HT clearance metabolism and ameliorating redox reactions. Our results may provide evidence for an indication of PFO closure in migraine and support the related potential mechanism. KEY POINTS: Aura, and levels of cystatin-C and calcium are biomarkers of migraineurs with a patent foramen ovale (PFO). The clearance of pulmonary metabolism of 5-HT and deoxygenated blood might be the reason for the improvement of migraine symptoms in patients with PFO. The posterior region of the brain is the main area responsible for PFO-induced migraine.

Effects of Dietary Supplementation with Lactobacillus reuteri Postbiotics on Growth Performance, Intestinal Flora Structure and Plasma Metabolome of Weaned Piglets.

Probiotics and their postbiotics have the potential to improve the health and growth performance of piglets, which has brought them widespread attention in the post-antibiotic era. In the present study, the effects of dietary supplementation of Lactobacillus reuteri postbiotics on the growth performance, intestinal flora structure and plasma metabolome of weaned piglets were investigated. A total of 816 healthy male piglets with uniform weight were divided into two treatment groups: piglets in the control (CTR) group were fed with a basic diet, and the ones in the LAC group were fed with the basic diet supplemented with 500 mg/kg Lactobacillus reuteri postbiotics. There were six replicates in each group and 68 piglets in each replicate. The animal trial lasted for 30 days. The feces and blood of piglets were collected for investigation, and the growth performance during the trial was counted. Our outcomes show that dietary supplementation with Lactobacillus reuteri postbiotics had no effect on the growth performance of piglets; however, it reduced the mortality rate of piglets by 6.37%. The levels of total superoxide dismutase in the serum, propionic acid and butyric acid in the feces were elevated, and the content of malondialdehyde in the serum was decreased with Lactobacillus reuteri postbiotics-treated piglets (p < 0.05). The fecal flora sequencing results show that the relative abundance of Firmicutes and monoglobus was upregulated, and the relative abundance of Bacteroides was downregulated with Lactobacillus reuteri postbiotics-treated piglets (p < 0.05). In addition, the levels of propionic acid and butyric acid in the feces were positively correlated with the relative abundance of Firmicutes and negatively correlated with the relative abundance of Bacteroides (p < 0.05). The plasma metabolome results show that dietary supplementation with Lactobacillus reuteri postbiotics raised the level of coenzyme Q10 in the serum, and the abundance of coenzyme Q10 was positively correlated with the relative abundance of Firmicutes and the level of total superoxide dismutase in the serum. In conclusion, dietary supplementation with Lactobacillus reuteri postbiotics contributed to improving the antioxidant function and reducing the mortality of piglets by regulating the structure of intestinal flora and upregulating the content of coenzyme Q10 in serum.

Proteomic, metabolomic and lipidomic profiles in community acquired pneumonia for differentiating viral and bacterial infections

Community-acquired pneumonia (CAP) has a significant impact on public health, especially in light of the recent SARS-CoV-2 pandemic. To enhance disease characterization and improve understanding of the underlying mechanisms, a comprehensive analysis of the plasma lipidome, metabolome and proteome was conducted in patients with viral and bacterial CAP infections, including those induced by SARS-CoV-2. Lipidomic, metabolomic and proteomic profiling were conducted on plasma samples of 69 patients suffering either from viral or bacterial CAP. Lipid and metabolite analyses were LC-MS-based, while proteomic analyses were performed using multiple panels of the Olink platform. Statistical methods, machine learning and pathway analyses were conducted investigating differences between the infection types. Through comparison of the bacterial and viral pathogen groups, distinct signatures were observed in the plasma profiles. Notably, linoleic acid-derived inflammation signaling metabolites (EpOME and DiHOME) were increased in viral CAP compared to bacterial CAP. Similarly, proteins involved in cellular immune response and apoptosis (LAG-3 and TRAIL) showed elevated levels in viral CAP, while bacterial CAP exhibited notable elevation in pattern-recognizing receptors (CLEC4D and EN-RAGE). Additionally, within the lipidomic profile at baseline, several lipids displayed notable differences between viral and bacterial pneumonia, including bile acids (GCA, TCA, TCDCA), various tri- and diglycerides (TGs and DGs), and several phosphatidylcholines (PCs). These findings hold promise for facilitating the differential diagnosis of viral and bacterial pulmonary infections based on the systemic lipidome, metabolome and proteome, enabling timely treatment decisions. Additionally, they highlight potential targets for drug research, advancing therapeutic interventions in CAP. By providing valuable insights into the molecular characterization of CAP, this study contributes to the improvement of understanding the disease and, ultimately, the development of effective treatment strategies.

Plasma metabolome reveals altered oxidative stress, inflammation, and amino acid metabolism in dogs with idiopathic epilepsy.

Idiopathic epilepsy (IE) is the most common chronic neurological disease in dogs and an established natural animal model for human epilepsy types with genetic and unknown etiology. However, the metabolic pathways underlying IE remain largely unknown. Plasma samples of healthy dogs (n = 39) and dogs with IE (n = 49) were metabolically profiled (n = 121 known target metabolites) and fingerprinted (n = 1825 untargeted features) using liquid chromatography coupled to mass spectrometry. Dogs with IE were classified as mild phenotype (MP; n = 22) or drug-resistant (DR; n = 27). All dogs received the same standard adult maintenance diet for a minimum of 20 days (35 ± 11 days) before sampling. Data were analyzed using a combination of univariate (one-way analysis of variance or Kruskal-Wallis rank sum test), multivariate (limma, orthogonal partial least squares-discriminant analysis), and pathway enrichment statistical analysis. In dogs with both DR and MP IE, a distinct plasma metabolic profile and fingerprint compared to healthy dogs was observed. Metabolic pathways involved in these alterations included oxidative stress, inflammation, and amino acid metabolism. Moreover, significantly lower plasma concentrations of vitamin B6 were found in MP (p = .001) and DR (p = .005) compared to healthy dogs. Our data provide new insights into the metabolic pathways underlying IE in dogs, further substantiating its potential as a natural animal model for humans with epilepsy, reflected by related metabolic changes in oxidative stress metabolites and vitamin B6. Even more, several metabolites within the uncovered pathways offer promising therapeutic targets for the management of IE, primarily for dogs, and ultimately for humans.

Effects of acteoside from Cistanche tubulosa on the plasma metabolome of cancer-related fatigue mice inoculated with colon cancer cells.

To elucidate the metabolic mechanisms by which acteoside (ACT) isolated from Cistanche tubulosa alleviates cancer-related fatigue (CRF) in a murine model of colon cancer with cachexia. BALB/c mice inoculated with C26 colon cancer cells were treated with paclitaxel (PTX, 10mg/kg) and ACT (100mg/kg) alone or in combination for 21days. Fatigue-associated behaviors, tumor inhibition rate, and skeletal muscle morphology assessed by hematoxylin-eosin (H&E) staining and electron microscopy were evaluated. Finally, liquid chromatography-mass spectrometry (LC/MS) was employed to investigate alterations in the plasma metabolic profile of tumor-bearing mice with CRF in response to ACT treatment, and the affinity between metabolite-associated proteins and ACT was verified by Surface plasmon resonance (SPR) assay. Our study demonstrated the presence of CRF in the colon cancer mouse model, with the severity of fatigue increasing alongside tumor growth. Administration of ACT ameliorated both tumor burden and PTX-induced muscle fatigue-like behavior. LC/MS analysis identified a panel of differentially regulated metabolites, including trans-aconitine, citric acid, 3-coumaric acid, ephedrine, thymine, cytosine, indole-3-acetic acid, and pantothenol-9. These metabolites were primarily enriched in pathways associated with valine biosynthesis, tyrosine metabolism, tryptophan metabolism, and biosynthesis of pyridine alkaloids. Furthermore, several key enzymes, including CYP3A4, CYP19A1, CYP2E1, TNF, BCL-2, RYR2, and ATP2A1, were identified as potential targets underlying the anti-CRF effects of ACT. This study suggests that ACT derived from C. tubulosa harbors protective properties against cancer-related fatigue mediated by tumor cells.

Multi-omics integration and immune profiling identify possible causal networks leading to uterine microbiome dysbiosis in dairy cows that develop metritis

BackgroundCows that develop metritis experience dysbiosis of their uterine microbiome, where opportunistic pathogens overtake uterine commensals. An effective immune response is critical for maintaining uterine health. Nonetheless, periparturient cows experience immune dysregulation, which seems to be intensified by prepartum over-condition. Herein, Bayesian networks were applied to investigate the directional correlations between prepartum body weight (BW), BW loss, pre- and postpartum systemic immune profiling and plasma metabolome, and postpartum uterine metabolome and microbiome.ResultsThe Bayesian network analysis showed a positive directional correlation between prepartum BW, prepartum BW loss, and plasma fatty acids at parturition, suggesting that heavier cows were in lower energy balance than lighter cows. There was a positive directional correlation between prepartum BW, prepartum systemic leukocyte death, immune activation, systemic inflammation, and metabolomic changes associated with oxidative stress prepartum and at parturition. Immune activation and systemic inflammation were characterized by increased proportion of circulating polymorphonuclear cells (PMN) prepartum, B-cell activation at parturition, interleukin-8 prepartum and at parturition, and interleukin-1β at parturition. These immune changes together with plasma fatty acids at parturition had a positive directional correlation with PMN extravasation postpartum, which had a positive directional correlation with uterine metabolites associated with tissue damage. These results suggest that excessive PMN migration to the uterus leads to excessive endometrial damage. The aforementioned changes had a positive directional correlation with Fusobacterium, Porphyromonas, and Bacteroides in cows that developed metritis, suggesting that excessive tissue damage may disrupt physical barriers or increase substrate availability for bacterial growth.ConclusionsThis work provides robust mechanistic hypotheses for how prepartum BW may impact peripartum immune and metabolic profiles, which may lead to uterine opportunistic pathogens overgrowth and metritis development.