Metabolomics Research Articles

Insights into IAV Replication and Lipid Metabolism in Suspension-Adapted MDCK-STAT1-KO Cells

Objectives: The industrial production of influenza vaccines is facing significant challenges, particularly in improving virus production efficiency. Despite advances in cell culture technologies, our understanding of the production characteristics of high-yield suspension cell lines remains limited, thereby impeding the development of efficient vaccine production platforms. This study aims to investigate the key features of STAT1 knockout suspension-adapted MDCK cells (susMDCK-STAT1-KO) in enhancing influenza A virus (IAV) production. Methods: Suspension-adapted susMDCK-STAT1-KO cells were compared to suspension-adapted wild-type MDCK cells (susMDCK) for IAV production. Virus quantification, gene expression analysis, and cholesterol deprivation assays were performed. Metabolite profiles, viral RNA quantification, and lipid and dry weight measurements were also conducted to assess the viral replication and release efficiency. Results: The susMDCK-STAT1-KO cells exhibited significantly improved virus adsorption (64%) and entry efficiency (75%) for the H1N1 virus, as well as accelerated viral transcription and replication for both the H1N1 and H9N2 viruses. Virus release was identified as a limiting factor, with a 100-fold higher intracellular-to-extracellular viral RNA ratio. However, the STAT1-KO cells showed a 2.39-fold higher release rate (750 virions/cell/h) and 3.26-fold greater RNA release for the H1N1 virus compared to wild-type cells. A gene expression analysis revealed enhanced lipid metabolism, particularly cholesterol synthesis, as a key factor in viral replication and release. Cholesterol deprivation resulted in reduced viral titers, confirming the critical role of intracellular cholesterol in IAV production. Conclusions: This study demonstrates the enhanced influenza virus production capacity of susMDCK-STAT1-KO cells, with significant improvements in viral yield, replication, and release efficiency. The findings highlight the importance of STAT1-mediated immune modulation and cholesterol metabolism in optimizing virus production. These insights provide a foundation for the development of more efficient vaccine production platforms, with implications for large-scale industrial applications.

OP19 Pre- and Post-diagnostic Metabolomic Biomarker Profiling of Over 700,000 Individuals in Three National Biobanks Enables Prediction of Inflammatory Bowel Disease Onset and Complications

Abstract Background We currently lack sensitive and specific prognostic biomarkers of inflammatory bowel disease (IBD) onset and severity. In three large, population-scale cohorts, we investigated metabolomic biomarkers for predicting disease onset and course. Methods We measured 250 metabolomic biomarkers using nuclear magnetic resonance (NMR) in ~500,000 individuals from the UK Biobank [1]. 2036 individuals had prevalent Crohn’s disease (CD) and 4029 had ulcerative colitis (UC) at recruitment. 1242 participants developed incident CD and 2283 incident UC during 10-year follow-up, allowing us to study the pre-diagnostic metabolic profile of IBD. We investigated associations between individual metabolomic biomarkers with prevalent and incident IBD using logistic and proportional hazards regressions. Among the incident IBD cohort we used LASSO with five-fold cross-validation on 50% of the cohort to build predictive models of future IBD onset. The resulting predictive model was validated in the Estonian Biobank and the Finnish THL Biobank (N ~218,000), confirming its robustness in multiple cohorts. Finally, in the UK Biobank we identified individuals with prevalent IBD who had IBD-related emergency admission, surgery or death and used these to develop a disease course score. Results GlycA, a biomarker of systemic inflammation, was the strongest association for both incident and prevalent UC and CD. We also identified associations between non-inflammatory metabolomic markers and IBD, such as albumin, branched-chain amino acids, cardiovascular lipid factors, fatty acids and creatinine. Notably, the perturbation in metabolites was markedly greater in CD versus UC, showing that CD is a more systemic illness (Fig. 1). Metabolomics scores were predictive of CD (AUC=0.69) and UC (0.62) onset up to 10 years prior to diagnosis, with performance for CD improving closer to diagnosis (2y follow-up AUC: CD 0.76, UC 0.62). Patients in the highest decile of incident metabolomic score were substantially more likely to develop the disease, with this observation being consistent in other two biobanks (Fig. 2). Metabolite profiles were predictive of IBD complications in prevalent patients (AUC 0.68 for CD; 0.62 for UC), outperforming both genetic and blood biomarker scores. Conclusion Leveraging cohort-wide profiling of population-scale biobanks, we present the largest metabolomics study and the largest pre-diagnostic study of IBD to date. Pre-diagnostic metabolomic scores are predictive of disease onset. These results warrant further investigation of multi-omic prediction of the onset, comorbidities and disease course of IBD. References * A.S, K.S. contributed equally. # T.J., J.C.B., C.L. contributed equally. [1] Nightingale Health Biobank Collaborative Group, Barrett, J. C., Esko, T., Fischer, K., Jostins-Dean, L., Jousilahti, P., ... & Estonian Biobank Research Team. (2023). Metabolomic and genomic prediction of common diseases in 477,706 participants in three national biobanks. medRxiv, 2023-06. Figure 1: Associations between clinically validated biomarkers and prevalent IBD Figure 2: UK Biobank-derived score consistency across the three biobanks

DOP121 Distinct tryptophan metabotypes associate with disease activity, treatment escalation and future disease progression in Inflammatory Bowel Disease

Abstract Background The kynurenine pathway is upregulated in inflammatory bowel disease (IBD) in a disease-activity induced manner. This route of tryptophan (Trp) degradation leads to a variety of bioactive compounds, including several with anti-inflammatory (e.g., kynurenic acid (Kyna), xanthurenic acid (Xana)) or cytotoxic (quinolinic acid (Quin)) effects. Our aim was to stratify patients with IBD into distinct subgroups based on their Trp metabolic profiles (Trp metabotypes) and to assess their prognostic capacity regarding disease progression. Methods We measured 16 Trp-related metabolites with particular attention to kynurenine pathway metabolites in the serum of a longitudinal German cohort (hypothesis generation) of 82 Ulcerative Colitis (UC) and 52 Crohn’s Disease (CD) patients. We used unsupervised k-means clustering to identify distinct clusters of patients based on their metabolite profiles and employed regression analysis to identify associations between the metabotypes and disease activity and progression. The analysis was expanded to three independent prospective validation cohorts recruited in France (n = 281 UC, 788 CD), the USA (n = 329 controls, 470 UC, 374 CD) and an inception cohort in Norway (n = 179 UC, 340 CD). Results We found four highly stable metabotypes within the discovery cohort (Fig 1A). Strong signals from the anti-inflammatory compounds Kyna and Xana characterized the first two metabotypes (High/Low Kyna metabotypes), which exhibited opposing associations with clinical, biochemical, and endoscopic disease activity. The third is characterized by high Quin levels and increased disease activity, while the fourth is metabolically heterogeneous and has lower disease activity (Fig 1B). These metabotypes were present in both UC and CD, albeit with differing proportions. Associations with disease activity were confirmed in three validation cohorts. Furthermore, we found differential probabilities between the metabotypes regarding future escalation to biologic therapies, requirement for steroid courses, IBD-related hospitalization and surgery. This was particularly evident when comparing individuals in the Low Kyna group with the lower incidence rates observed in the High Kyna group (Fig. 1C). Conclusion A distinct pattern of Trp degradation occurs in a subset of IBD patients that is strongly and consistently linked with disease activity and longitudinal outcomes of disease progression across multiple recruitment sites spanning two continents. Further work will focus on metabolic characterization of the metabotypes to understand the physiological processes driving the differentiation. This work may lead to the design of new therapeutic modalities targeting Trp metabolism in distinct patient groups.

Distinct metabolic profiles of key metabolites in sugarcane leaves and culms highlight their potential roles in growth and development

Distinct metabolic profiles of key metabolites in sugarcane leaves and culms highlight their potential roles in growth and development

Integrated multi-omics reveals the Bacillus amyloliquefaciens BA40 against Clostridium perfringens infection in weaned piglets.

Clostridium perfringens (C. perfringens) can cause necrotic enteritis and higher mortality rates in piglets, by impairing the intestinal barrier function. Bacillus amyloliquefaciens 40 (BA40) has showed potential ability to reduce C. perfringens infections, but the mechanisms responsible for its effectiveness remain unclear. This study aims to evaluate the impact of BA40 on inflammation induced by C. perfringens and to explain the mechanisms underlying its therapeutic effects. We aim to show how BA40 can bolster piglet health by strengthening the intestinal barrier and regulating immune responses. We used piglets and cellular models, alongside microbiomics, metabolomic, and transcriptomic analyses, to investigate BA40's impact on C. perfringens-induced inflammation. A model of C. perfringens infection was constructed using piglets and cells to investigate the effect of BA40 on its phenotype. Microbiomics, metabolomics, and transcriptomics analyses were subsequently used to investigate the mechanisms of protection and immune response to BA40 on the intestinal barrier of piglets. Our study revealed significant improvements in piglet health following BA40 administration. Notably, BA40 strengthened the intestinal mucosal barrier and mitigated the inflammatory response triggered by C. perfringens BA40 decreased harmful bacteria and increased beneficial bacteria. Metabolite profiles improved, showing a reduction in harmful substances. Transscriptomics analysis indicated BA40's role in TNF/NF-κB signaling pathway, hinting at its ability to regulate immune responses and reduce intestinal inflammation. Cellular assays further confirmed BA40's capacity to diminish inflammatory cytokine release and encourage the differentiation of anti-inflammatory macrophages. Datasets from the present study demonstrate that BA40 modulates gut microbes and metabolites, inhibits inflammation-related signaling pathways, and maintains gut barrier function. Our findings not only deepen our understanding of the therapeutic capacity of BA40 but also provide a theoretical foundation for the development of probiotics and alternative therapies aimed at improving piglet gut health.

Exploring chitin metabolite profiles and sensitivity differences in Collembola species exposed to teflubenzuron.

Effective environmental risk assessments of chemical plant protection products, such as benzoylurea pesticides, are crucial for safeguarding ecosystems. These pesticides, including teflubenzuron, target chitin synthesis in arthropods but also pose risks to non-target soil fauna like Collembola, which play essential roles in decomposition and nutrient cycling. This study combines traditional toxicity tests with a metabolomic approach to examine the interspecies specific sensitivity of three Collembola species - Sinella curviseta, Ceratophysella denticulata, and Folsomia candida - to teflubenzuron. The investigation focused on reproduction, bioaccumulation, and changes in chitin-related metabolites as indicators of pesticide impacts. Results revealed significant interspecies specific variability in sensitivity, with F. candida showing higher susceptibility towards teflubenzuron, possibly due to greater bioaccumulation factors. Metabolomic analysis highlighted distinct patterns in chitin metabolite alterations among the species, correlating with their differential sensitivity. Notably, metabolites like trehalose and glucose, crucial for chitin synthesis, were significantly affected by teflubenzuron within seven days of exposure. Despite high soil concentrations of the pesticide, S. curviseta demonstrated resilience in traditional life-history endpoints, such as reproduction and survival. However, metabolomics indicated a biochemical response to even low internal concentrations of teflubenzuron, underscoring the complexity of their interactions with environmental stressors. This study emphasizes the importance of incorporating metabolomics to understand the differential responses of non-target organisms to pesticides and advocates for species-specific risk assessments in pesticide regulation. The distinct metabolic responses among Collembola species to chitin synthesis inhibitors provide critical insights into their ecological resilience or vulnerability, enhancing our understanding of ecosystem dynamics and the potential ramifications of chemical exposure.

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.

Exploratory profiling of metabolites in cerebrospinal fluid using a commercially available targeted LC-MS based metabolomics kit to discriminate leptomeningeal metastasis

BackgroundLeptomeningeal metastasis (LM) is a devastating complication of cancer that is difficult to treat. Thus, early diagnosis is essential for LM patients. However, cerebrospinal fluid (CSF) cytology has low sensitivity, and imaging approaches are ineffective. We explored targeted CSF metabolic profiling to discriminate among LM and other conditions affecting the central nervous system (CNS).MethodsWe quantitatively measured amino acids, biogenic amines, hexoses, acylcarnitines (AC), cholesteryl esters (CE), glycerides, phosphatidylcholines (PC), lysophosphatidylcholines (LPC), sphingomyelins (SM), and ceramides (Cer) in 117 CSF samples from various groups of healthy controls (HC, n = 10), patients with LM (LM, n = 47), parenchymal brain tumor (PBT, n = 45), and inflammatory disease (ID, n = 13) with internal standards using the Absolute IDQ- p400® targeted mass spectrometry kit. Metabolites detected in > 90% of samples or showing a difference in proportional level between groups ≥ 75% were used in logistic regression models when there was no single metabolite with AUC = 1 for the groups of comparison.ResultsPC and SM had higher levels in LM than in PBT or HC, whereas LPC had lower level in PBT than the other groups. Glycerides and Cer levels were higher in PBT and LM than in HC. Long-chain AC level in PBT was lower than in LM or HC. A regression model including Ala, PC (42:7), PC (30:3), PC (37:0), and Tyr achieved complete discrimination (AUC = 1.0) between LM and HC. In comparison of PBT and HC, twenty-six individual metabolites allowed complete discrimination between two groups, and between ID and HC fourty-six individual lipid metabolites allowed complete discrimination. Twenty-one individual metabolites (18 ACs and 3 PCs) allowed complete discrimination between LM and PBT.ConclusionsUsing a commercial targeted liquid chromatography-mass spectrometry (LC-MS) metabolomics kit, we were able to differentiate LM from HC and PBT. Most of the discriminative metabolites among different diseases were lipid metabolites, for which their CNS distribution and quantification in different cell types are largely unknown, whereas amino acids, biogenic amines, and hexoses failed to show significant differences. Future validation studies with larger, controlled cohorts should be performed, and hopefully, the kit may expand its metabolite coverage for unique cancer cell glucose metabolism.

Comprehensive analysis of Leonotis nepetifolia flower extracts: phytochemical composition and toxicity in zebrafish embryos

Leonotis nepetifolia (L.) R. Br., a plant used in traditional medicine, has underexplored phytochemical and toxicological profiles. This study investigates the metabolite profile of L. nepetifolia flowers and assesses their toxicity using zebrafish (ZF) embryos. The main active compounds were characterised using metabolomic approaches. ZF embryos were exposed to methanol extract (CEF), n-hexane (FHF) and ethyl acetate (FAF) fractions at different concentrations for 96 h. Toxicological effects were assessed including acetylcholinesterase activity, lipid peroxidation, cardiotoxicity, as well as hatching delay, developmental defects and morphological malformations. Phytochemical analysis revealed diverse metabolites, including phytosterols, terpenoids, flavonoids and phenylpropanoids. Verbascoside, a major compound, was isolated from the flowers for the first time. Toxicological assessments showed that CEF and FAF caused various toxic effects, with FAF showing pronounced embryotoxic and teratogenic effects. This study highlights the chemical diversity and potential toxicological risks of L. nepetifolia, emphasising the need for thorough evaluations of herbal medicines.

The roles of IDH2 and glutathione metabolism in cetuximab resistance in head and neck squamous cell carcinoma investigated by metabolomics and transcriptomics.

Cetuximab resistance is a significant challenge in the treatment of head and neck squamous cell carcinoma (HNSCC). In this study, cetuximab-resistant HNSCC cell lines were established, and untargeted metabolomics was used to detect differences in metabolite profiles between sensitive and resistant cell lines. It was found that glutathione metabolism significantly differed between the sensitive and resistant lines. Combining these findings with transcriptome data, correlation analysis of metabolites revealed that IDH2 regulated glutathione metabolism and contributed to cetuximab resistance in FaDu cells. In vitro experiments showed that IDH2 was highly expressed in FaDu-CR cells, and IDH2 knockdown significantly enhanced the sensitivity of FaDu and FaDu-CR cells to cetuximab. IDH2 knockdown reduced GSH levels and GPX4 expression in FaDu and FaDu-CR cells under cetuximab treatment, while increasing lipid ROS levels. In vivo experiments demonstrated that IDH2 knockdown decreased the tumorigenic ability of FaDu-CR cells in nude mice treated with cetuximab, as well as reduced GPX4 and Ki67 levels in tumor tissues. In conclusion, IDH2 regulated glutathione metabolism and contributed to cetuximab resistance in HNSCC. This study explores strategies to ameliorate cetuximab resistance in HNSCC preclinical models, providing new insights for reversing cetuximab resistance in HNSCC.

Relationship Between the Salivary Microbiome and Oral Malodor Metabolites in Older Thai Individuals with Periodontitis and the Cytotoxic Effects of Malodor Compounds on Human Oral Squamous Carcinoma (HSC-4) Cells.

Background/Objectives: Halitosis is primarily caused by the activity of oral microorganisms. In this study, we employed metagenomic sequencing and metabolomic approaches to investigate the differences in salivary microbiota and metabolite profiles between individuals with halitosis and periodontitis and healthy controls. Additionally, we expanded the study to examine how oral malodorous compounds interact with human oral squamous carcinoma (HSC-4) cells. Methods: Saliva samples were collected and analyzed using Ultra-High Performance Liquid Chromatography-Mass Spectrometry (UHPLC-MS) to identify metabolites. We then assessed the correlations between the microbiota and metabolites. Furthermore, the impact of oral malodorous substances on HSC-4 cells was investigated by evaluating apoptosis, antioxidant activity, and inflammatory properties. Results: The microbiota and metabolite profiles showed significant differences between the halitosis with periodontitis group and the periodontally healthy group. The halitosis with periodontitis group exhibited significantly higher relative abundances of eight genera: Tannerella, Selenomonas, Bacteroides, Filifactor, Phocaeicola, Fretibacterium, Eubacterium saphenum, and Desulfobulbus. In contrast, the periodontally healthy group showed significantly higher relative abundances of Family XIII UCG-001, Haemophilus, and Streptobacillus. Two metabolites, 2,3-dihydro-1H-indole and 10,11-dihydro-12R-hydroxy-leukotriene E4, were significantly higher in individuals with halitosis and periodontitis. In the treatment of HSC-4 cells with metabolites, dimethyl sulfide (DMS) did not show significant effects while indole appeared to induce cell death in HSC-4 cells by triggering apoptotic pathways. Additionally, both indole and DMS affected the inflammatory and antioxidant properties of HSC-4 cells. Conclusions: This study provides insights into the mechanisms of halitosis by exploring the correlations between microbiota and metabolite profiles. Furthermore, oral metabolites were shown to impact the cellular response of HSC-4 cells.

Profiling of metabolites and minerals from black ginger and blue turmeric rhizomes by gas chromatography–mass spectrometry analysis and their biopotentials

Profiling of metabolites and minerals from black ginger and blue turmeric rhizomes by gas chromatography–mass spectrometry analysis and their biopotentials

Supplementation with Complex Phytonutrients Enhances Rumen Barrier Function and Growth Performance of Lambs by Regulating Rumen Microbiome and Metabolome.

Complex phytonutrients (CPS) have attracted extensive interest due to their anti-inflammatory effects. This investigation focused on the impact of CPS on rumen health in lambs on high-concentrate diets, emphasizing growth performance, ruminal fermentation, epithelial barrier integrity, ruminal metabolism, and microbial communities. A total of 54 lambs, 3 months old and with a 30.42 ± 0.54 kg body weight, were randomly assigned to three treatment groups, with six replicates per treatment and three lambs per replicate. The lambs received a basal diet (the ratio of concentrate to forage was 75:25) without CPS supplementation (CON) or with the inclusion of 2.5 g/kg (CPS2.5) or 5.0 g/kg CPS (CPS5.0) for a total of 60 days. The CPS groups exhibited increased growth performance and improved rumen fermentation parameters. Mechanistically, CPS enhanced rumen epithelial barrier function, thereby lowering inflammation and inhibiting the overactivation of the JNK/p38 MAPK signaling pathway, and the effect of CPS5.0 was better than that of CPS2.5. Notably, CPS5.0 could optimize the composition of rumen microbiota and increase the levels of Ursolic acid and other metabolites. The strong associations between rumen bacteria and health-related indicators and differential metabolites were further highlighted. Our findings suggest that adding CPS to lambs' diets has widespread positive impacts, including improved growth performance, reduced inflammation and mRNA relative expression of apoptosis-related genes, enhanced barrier function, and beneficial changes in the rumen microbiome and metabolite profiles.

Structural alterations in butyrylated and recrystallized high-amylose maize starch and their effect on gut microbiota during in vitro fermentation.

In this study, type-3 resistant starch (RS) with enhanced thermal stability and excellent short-chain fatty acid (SCFA) production was obtained through the butyrylation and subsequent recrystallization at 4 °C of high-amylose maize starch (HAMS). We comprehensively examined and contrasted the structural attributes and in vitro human fecal fermentation behavior of butyrylated RS (BRS) with varying degrees of substitution. Fourier-transform infrared analysis validated the successful integration of carbonyl groups into the starch matrix. This phenomenon was evident through the characteristic peaks at 1727 cm-1. X-ray diffraction and thermal stability analyses delineated the distinct B-type crystalline structure and high relative crystallinity of 25.31 % and 22.23 % of the 10 % and 15 % butyric anhydride BRS-10 and BRS-15, respectively, which differed from that of HAMS. Their second peak gelatinization temperature values reached 95.6 °C and 92.6 °C. The in vitro fermentation of BRS fostered SCFA production, boosting the relative abundance of beneficial bacteria (e.g., Ligilactobacillus and Roseburia). Meanwhile, the extensively modified BRS favored the propagation of Faecalibacterium and Bifidobacterium, maintaining intestinal microbiota advantage. These findings underscore the significant effects of butyrylation modification on fermentation kinetics, metabolite profiles, and gut microbiota composition, providing invaluable insights into the development of functional food products that aim to bolster colonic health.

Amino acids and CART distinguish A-β+ Ketosis-Prone Diabetes from type 1 and type 2 diabetes during hyperglycemic crises.

When clinically stable, patients with A-β+ Ketosis-Prone Diabetes (KPD) manifest unique markers of amino acid metabolism. Biomarkers differentiating KPD from type 1 (T1D) and type 2 diabetes (T2D) during hyperglycemic crises would accelerate diagnosis and management. Compare serum metabolomics of KPD, T1D and T2D patients during hyperglycemic crises, and utilize Classification and Regression Tree (CART) modeling to distinguish these forms of diabetes. Urban hospital emergency center. Adults with KPD, T1D and T2D during hyperglycemic crises (with or without diabetic ketoacidosis (DKA), and healthy controls. Comparisons of serum metabolite and hormonal profiles, and CART analysis. Group differences in concentrations of amino acids, their metabolites and relationship to glucose counterregulatory hormones; C-peptide cutoffs and analytes to distinguish KPD, T1D and T2D. Concentrations of most amino acids were similar in KPD and T1D and lower compared to T2D (P<0.05). Glucagon and cortisol concentrations were correlated with 3-methylhistidine and blood urea nitrogen in KPD but not in T1D. A C-peptide cutoff of 0.496 ng/mL differentiated T1D from KPD during DKA. CART revealed that a regression model based on the concentrations of β-hydroxybutyrate, C-peptide, glucagon, alpha-keto-β-methylvalerate, cystine and myristoyl-L-carnitine distinguished KPD from T1D and T2D. During DKA, KPD and T1D patients have similarly altered amino acid profiles that differentiate them from T2D patients. Elevated protein catabolic hormones drive altered amino acid metabolism in KPD, rather than insulin deficiency as with T1D. A combination of 6 analytes differentiates KPD from T1D and T2D during hyperglycemic crises.

In vitro anti-diabetic potential of medicinal herbs commonly used in the Ayurvedic system of Sri Lanka with comprehensive metabolite profiling of Phyllanthus emblica using GC-MS and LC-HRMS

This study assessed the in vitro anti-diabetic potential and bioactive constituents of ten Sri Lankan medicinal herbs. Initial screening of aqueous extracts for starch-digesting enzyme inhibition prioritised three plants with notable activity (p ≤ 0.05), for further assessment using methanolic extracts: Phyllanthus emblica (PE), Cassia auriculata (CA), and Hemidesmus indicus (HI). The selected plants were tested for starch-digesting enzyme inhibition, cytotoxicity, and bioactive metabolite identification, with PE subjected to GC-MS and LC-HRMS analyses. All three extracts contained alkaloids, flavonoids, tannins, and terpenoids, except saponins and steroids in PE. GC-MS analysis of PE annotated ten compounds, eight with anti-diabetic properties, while LC-HRMS annotated thirty metabolites, including fourteen anti-diabetic compounds. Cell viability assessments confirmed the non-toxic nature of PE, CA, and HI. The significant enzyme inhibition and non-toxic nature of PE highlight its potential to treat type 2 diabetes. Further in vivo and clinical studies are essential to determining effective dosage and toxicity levels.

Exploring Hornstedtia scyphifera: an extensive multimethod phytochemical investigation reveals the chemical composition and bioactive potential

Abstract Hornstedtia scyphifera (J.Koenig) Steud. represents a lesser-known member of the ginger family (Zingiberaceae) that is used in Malaysia as spice and traditional medicine. The phytochemical investigation of leaves from this species utilizing diverse analytical methods has provided comprehensive insights into its chemical profile for the first time. Headspace gas chromatography-mass spectrometry (HS-GCMS) and GCMS analyses of essential oil and nonpolar extracts verified α-pinene, camphene, p-cymene, and camphor as main volatile compounds. Metabolite profiling of the crude extract by ultra-high-performance-liquid chromatography-high resolution mass spectrometry (UHPLC-HRMS) unveiled terpenoids, flavonoids and other phenolics as major compound classes. Isolation and follow-up structure elucidation, involving 1D and 2D NMR, HRMS, UV and CD analysis, yielded two new sesquiterpenoids, (1R,5S,6S,7R,10R)-mustak-14-oic acid (1) and (1R,6S,7S,10R)-6-hydroxy-anhuienosol (2), along with 24 known compounds (seven terpenoids, seven flavonoids, ten phenolics), 21 of these never reported for H. scyphifera. Additionally, the crude extract and fractions from the purification process were screened for antibacterial and antifungal activity. This is supplemented by an extensive literature research for described bioactivities of all isolated compounds. Our results support and explain previously detected antimicrobial, antifungal and neuroprotective effects of H. scyphifera extracts and provide evidence for its potential pharmacological importance.

Comparative metabolomic analysis of Haematococcus pluvialis during hyperaccumulation of astaxanthin under the high salinity and nitrogen deficiency conditions.

Revealing the differences of metabolite profiles of H. pluvialis during hyperaccumulation of astaxanthin under the high salinity and nitrogen deficiency conditions was the key issues of the present study. To investigate the optimum NaCl and NaNO3 concentration and the corresponding metabolic characteristic related to the astaxanthin accumulation in H. pluvialis, a batch culture experiment was conducted. The results indicated that 7.5g·L- 1 and 0g·L- 1 (nitrogen deficiency) were the optimum NaCl and NaNO3 levels for the astaxanthin accumulation respectively, under which the highest astaxanthin contents reached up to 7.51mg·L- 1 and 5.60mg·L- 1. A total of 132 metabolites were identified using LC-MS/MS technique, among which 30 differential metabolites with statistical significance were highlighted. Subsequently, 18 and 10 differential metabolic pathways in the high salinity (HS) and nitrogen-deficient (ND) treatments were extracted and annotated respectively. The values of Fv/Fm, Yield and NPQ were all at the highest level in the ND group during the experiment. The levels of the metabolites in the ND group were almost lower than those both in the control (CK) and HS group, while which in the HS group were substantially at the higher or close levels compared to the CK group. Finally, 7 metabolic markers related to the astaxanthin accumulation were highlighted in the HS and ND group respectively. L-Proline, L-Aspartate, Uridine 5'-monophosphate (UMP), Succinate, L-2-Hydroxygluterate, L-Valine and Inosine 5'-monophosphate (IMP) were identified as the metabolic markers in the HS group, whose fold change were 0.85, 4.14, 0.31, 0.66, 3.10, 1.32 and 0.30. Otherwise, the metabolic markers were Glyceric acid, Thymine, sn-Glycerol 3-phosphate, Glycine, Allantoic acid, L-Valine and IMP in the ND group, with the fold change 0.23, 2.11, 0.38, 0.41, 0.50 and 2.96 respectively. The results provided the comparative metabolomic view of astaxanthin accumulation in H. pluvialis under the different cultivation conditions, moreover showed a novel insights into the astaxanthin production.

Metabolomic in severe traumatic brain injury: exploring primary, secondary injuries, diagnosis, and severity

BackgroundTraumatic brain injury (TBI) is a major public health concern worldwide, contributing to high rates of injury-related death and disability. Severe traumatic brain injury (sTBI), although it accounts for only 10% of all TBI cases, results in a mortality rate of 30–40% and a significant burden of disability in those that survive. This study explored the potential of metabolomics in the diagnosis of sTBI and explored the potential of metabolomics to examine probable primary and secondary brain injury in sTBI.MethodsSerum samples from 59 adult patients with sTBI and 35 age- and sex-matched orthopedic injury controls were subjected to quantitative metabolomics, including proton nuclear magnetic resonance (1H-NMR) and direct infusion/liquid chromatography-tandem mass spectrometry (DI/LC–MS/MS), to identify and quantify metabolites on days 1 and 4 post-injury. In addition, we used advanced analytical methods to discover metabo-patterns associated with sTBI diagnosis and those related to probable primary and secondary brain injury.ResultsOur results showed different serum metabolic profiles between sTBI and orthopedic injury (OI) controls, with significant changes in measured metabolites on day 1 and day 4 post-brain injury. The number of altered metabolites and the extent of their change were more pronounced on day 4 as compared to day 1 post-injury, suggesting an evolution of mechanisms from primary to secondary brain injury. Data showed high sensitivity and specificity in separating sTBI from OI controls for diagnosis. Energy-related metabolites such as glucose, pyruvate, lactate, mannose, and polyamine metabolism metabolites (spermine and putrescine), as well as increased acylcarnitines and sphingomyelins, occurred mainly on day 1 post-injury. Metabolites of neurotransmission, catecholamine, and excitotoxicity mechanisms such as glutamate, phenylalanine, tyrosine, and branched-chain amino acids (BCAAs) increased to a greater degree on day 4. Further, there was an association of multiple metabolites, including acylcarnitines (ACs), lysophosphatidylcholines (LysoPCs), glutamate, and phenylalanine, with injury severity at day 4, while lactate, glucose, and pyruvate correlated with injury severity on day 1.ConclusionThe results demonstrate that serum metabolomics has diagnostic potential for sTBI and may reflect molecular mechanisms of primary and secondary brain injuries when comparing metabolite profiles between day 1 and day 4 post-injury. These early changes in serum metabolites may provide insight into molecular pathways or mechanisms of primary injury and ongoing secondary injuries, revealing potential therapeutic targets for sTBI. This work also highlights the need for further research and validation of sTBI metabolite biomarkers in a larger cohort.

Changes in Microbial Community Diversity and the Formation Mechanism of Flavor Metabolites in Industrial-Scale Spontaneous Fermentation of Cabernet Sauvignon Wines.

The key flavor compound formation pathways resulting from indigenous microorganisms during the spontaneous fermentation of wine have not been thoroughly described. In this study, high-throughput metagenomic sequencing and untargeted metabolomics were utilized to investigate the evolution of microbial and metabolite profiles during spontaneous fermentation in industrial-scale wine production and to elucidate the formation mechanisms of key flavor compounds. Metabolome analysis showed that the total amount of esters, fatty acids, organic acids, aldehydes, terpenes, flavonoids, and non-flavonoids increased gradually during fermentation. Enrichment analysis indicated that metabolic pathways related to the synthesis, decomposition, transformation, and utilization of sugars, amino acids, and fatty acids were involved in the formation of key flavor compounds in wine. Metagenomic analysis revealed that Saccharomyces, Hanseniaspora, Zygosaccharomyces, Wickerhamiella, Lactobacillus, and Fructobacillus were the dominant taxa during spontaneous fermentation. They were significantly positively correlated with organic acids, fatty acids, esters, phenols, aldehydes, terpenes, and phenols. In conclusion, this research provides new insights into the metabolic pathways of key flavor compounds formed by indigenous microorganisms during wine fermentation.