PubMed

bioRxiv [Preprint]. 2024 Nov 3:2024.10.30.621159. doi: 10.1101/2024.10.30.621159.ABSTRACTThe cellular metabolism of macrophages depends on tissue niches and can control macrophage inflammatory or resolving phenotypes. Yet, the identity of signals within tissue niches that control macrophage metabolism is not well understood. Here, using single-cell RNA sequencing of macrophages in early mouse wounds, we find that, rather than gene expression of canonical inflammatory or resolving polarization markers, metabolic gene expression defines distinct populations of early wound macrophages. Single-cell secretomics and transcriptomics identify inflammatory and resolving cytokines expressed by early wound macrophages, and we show that these signals drive metabolic inputs and mitochondrial metabolism in an age-dependent manner. We show that aging alters the metabolome of early wound macrophages and rewires their metabolism from mitochondria to glycolysis. We further show that macrophage-derived Chi3l3 and IGF-1 can induce metabolic inputs and mitochondrial mass/metabolism in aged and bone marrow-derived macrophages. Together, these findings reveal that macrophage-derived signals drive the mitochondrial metabolism of macrophages within early wounds in an age-dependent manner and have implications for inflammatory diseases, chronic injuries, and age-related inflammatory diseases.IN BRIEF: This study reveals that macrophage subsets in early inflammatory stages of skin wound healing are defined by their metabolic profiles rather than polarization phenotype. Using single-cell secretomics, we establish key macrophage cytokines that comprise the in vivo wound niche and drive mitochondrial-based metabolism. Aging significantly alters macrophage heterogeneity and increases glycolytic metabolism, which can be restored to OxPHOS-based metabolism with young niche cytokines. These findings highlight the importance of the tissue niche in driving macrophage phenotypes, with implications for aging-related impairments in wound healing.HIGHLIGHTS: Single cell transcriptional analysis reveals that reveals that metabolic gene expression identifies distinct macrophage populations in early skin wounds.Single-cell secretomic data show that young macrophages contribute to the wound bed niche by secreting molecules such as IGF-1 and Chi3l3.Old wound macrophages display altered metabolomics, elevated glycolytic metabolism and glucose uptake, and reduced lipid uptake and mitochondrial mass/metabolism.Chi3l3 but not IGF-1 secretion is altered in macrophages in an age dependent manner.Chi3l3 can restore mitochondrial mass/metabolism in aged macrophages.PMID:39553941 | PMC:PMC11565841 | DOI:10.1101/2024.10.30.621159

J Thorac Dis. 2024 Oct 31;16(10):6936-6954. doi: 10.21037/jtd-24-1201. Epub 2024 Oct 14.ABSTRACTBACKGROUND: The gut microbiota and its associated metabolites play a critical role in shaping the systemic immune response and influencing the efficacy of immunotherapy. In this study, patients with extensive-stage small cell lung cancer (ES-SCLC) were included to explore the correlation between gut microbiota and metabolites and immunotherapy efficacy in patients with ES-SCLC.METHODS: Pre- and post-treatment, we collected stool samples from 49 ES-SCLC patients treated with an anti-programmed death-ligand 1 (PD-L1) antibody. We then applied 16S ribosomal RNA (rRNA) sequencing and liquid chromatography-mass spectrometry (LC-MS) non-targeted metabolomics technology. Subsequently, the gut microbiota and metabolites were identified and classified.RESULTS: The results showed no statistical difference in gut microbiota alpha and beta diversity between the responder (R) and non-responder (NR) patients at baseline. However, the alpha diversity of the R patients was significantly higher than that of the NR patients after treatment. There were also differences in the microbiome composition at the baseline and post-treatment. Notably, after treatment, Faecalibacterium, Clostridium_sensu_stricto_1, and [Ruminococcus]_torques were enriched in the R group, while Dubosiella, coriobacteriaceae_UCG-002 was enriched in the NR group. The non-targeted metabolomics results also indicated that short-chain fatty acids (SCFAs) were up-regulated in the R group after treatment. More, differential metabolites were enriched in the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways, including the PD-L1 expression and programmed death 1 (PD-1) checkpoint pathway in cancer.CONCLUSIONS: These findings are anticipated to provide novel markers for predicting the efficacy of immune checkpoint inhibitors (ICIs) in patients with ES-SCLC, and offer new directions for further research on molecular mechanisms.PMID:39552845 | PMC:PMC11565349 | DOI:10.21037/jtd-24-1201

Genes Dis. 2024 Feb 3;12(1):101239. doi: 10.1016/j.gendis.2024.101239. eCollection 2025 Jan.ABSTRACTIn precision cancer therapy, addressing intra-tumor heterogeneity poses a significant obstacle. Due to the heterogeneity of each cell subtype and between cells within the tumor, the sensitivity and resistance of different patients to targeted drugs, chemotherapy, etc., are inconsistent. Concerning a specific tumor type, many feasible treatments or combinations can be used by specifically targeting the tumor microenvironment. To solve this problem, it is necessary to further study the tumor microenvironment. Single-cell sequencing techniques can dissect distinct tumor cell populations by isolating cells and using statistical computational methods. This technology may assist in the selection of targeted combination therapy, and the obtained cell subset information is crucial for the rational application of targeted therapy. In this review, we summarized the research and application advances of single-cell sequencing technology in the tumor microenvironment, including the most commonly used single-cell genomic and transcriptomic sequencing, and their future development direction was proposed. The application of single-cell sequencing technology has been expanded to include epigenomics, proteomics, metabolomics, and microbiome analysis. The integration of these different omics approaches has significantly advanced the development of single-cell multiomics sequencing technology. This innovative approach holds immense potential for various fields, such as biological research and medical investigations. Finally, we discussed the advantages and disadvantages of using single-cell sequencing to explore the tumor microenvironment.PMID:39552788 | PMC:PMC11566696 | DOI:10.1016/j.gendis.2024.101239

Front Microbiol. 2024 Nov 1;15:1485936. doi: 10.3389/fmicb.2024.1485936. eCollection 2024.ABSTRACTBACKGROUND: Dysbiosis of gut microbiota (GM) is intricately linked with cognitive impairment and the incidence of traumatic brain injury (TBI) in both animal models and human subjects. However, there is limited understanding of the impact and mechanisms of fecal microbiota transplantation (FMT) on brain and gut barrier function in the treatment of TBI induced by gas explosion (GE).METHODS: We have employed FMT technology to establish models of gut microbiota dysbiosis in male rats, and subsequently conducted non-targeted metabolomics and microbiota diversity analysis to explore the bacteria with potential functional roles.RESULTS: Hematoxylin-eosin and transmission electron microscopy revealed that GE induced significant pathological damage and inflammation responses, as well as varying degrees of mitochondrial impairment in neuronal cells in the brains of rats, which was associated with cognitive decline. Furthermore, GE markedly elevated the levels of regulatory T cell (Tregs)-related factors interleukin-10, programmed death 1, and fork head box protein P3 in the brains of rats. Similar changes in these indicators were also observed in the colon; however, these alterations were reversed upon transfer of normal flora into the GE-exposed rats. Combined microbiome and metabolome analysis indicated up-regulation of Clostridium_T and Allobaculum, along with activation of fatty acid biosynthesis after FMT. Correlation network analysis indirectly suggested a causal relationship between FMT and alleviation of GE-induced TBI. FMT improved intestinal structure and up-regulated expression of tight junction proteins Claudin-1, Occludin, and ZO-1, potentially contributing to its protective effects on both brain and gut.CONCLUSION: Transplantation of gut microbiota from healthy rats significantly enhanced cognitive function in male rats with traumatic brain injury caused by a gas explosion, through the modulation of gut microbiome composition and the improvement of both gut and brain barrier integrity via the gut-brain axis. These findings may offer a scientific foundation for potential clinical interventions targeting gas explosion-induced TBI using FMT.PMID:39552646 | PMC:PMC11564976 | DOI:10.3389/fmicb.2024.1485936

Transfusion. 2024 Nov 17. doi: 10.1111/trf.18063. Online ahead of print.ABSTRACTBACKGROUND: Pathogen reduction technology (PRT)-treated apheresis platelets (APs) were returned without platelet swirl and with pH22°C < 6.2. The platelet donor was taking prescription levothyroxine and metformin plus over-the-counter medications and supplements. We hypothesized that either PRT or medication was causative.STUDY DESIGN AND METHODS: One AP from a double AP collection from this donor was PRT-treated, the other unit untreated. Units were assessed over 7-day storage with a standard panel of platelet assays and metabolomics using high resolution mass spectrometry. The dose effect of metformin on platelets over storage was evaluated in vitro using APs obtained from non-medicated donors.RESULTS: This donor's PRT- and non-PRT treated paired units had pH values <6.2 by the end of day 2. Lactate generation rates in the PRT- and non-PRT units were very high compared to previously reported values and approached that reported for anaerobic storage. Metabolomic analysis revealed impairments in a number of energy metabolic pathways between PRT- and non-PRT platelets; however, this did not support a major causative role of PRT in the significant upregulation of lactic acid production. Metformin caused a dose-dependent upregulation of glycolysis, resulting in pH decline.DISCUSSION: We conclude that metformin is the most likely cause for this donor's stored platelet pH failures. Metformin is commonly used to treat type 2 diabetes and is not a donor deferral medication. Further investigation is indicated into the potential impact of metformin on platelet storage characteristics, the potential implications for medication deferral, and the need for additional screening tools in the laboratory.PMID:39552144 | DOI:10.1111/trf.18063

Environ Sci Technol. 2024 Nov 17. doi: 10.1021/acs.est.4c05208. Online ahead of print.ABSTRACTThe extensive application of plastic products in daily human life has led to the accumulation of microplastics (MPs) in agricultural soil. However, little is known about the cross-generational toxicity of MPs on terrestrial invertebrates. In this study, two-generational Eisenia fetida was exposed to low-density polyethylene (LDPE, 0-5%, w/w) for 98 days to reveal the cross-generational toxicity and the underlying mechanisms. Results showed that LDPE-MPs not only perpetrated deleterious effects on the development, hatchability, and fecundity of the F0 generation but also stimulated the antioxidant defense activity, inhibited lipid peroxidation, and disordered neurotransmission in F1 generation individuals. The susceptibility of the epidermal-intestinal barrier to LDPE-MPs was dose-dependent. According to the transcriptomic analysis, the cross-generational earthworms confirmed significant perturbances in the cell cycle, neural activity-related pathways, and amino acid metabolism pathways (p < 0.05). Nevertheless, the metabolomic profile of F1 generation individuals exhibited significant hyperactive responses in glutathione metabolism and alanine, aspartate, and glutamate metabolism (p < 0.05). This study provides a comprehensive knowledge of LDPE-MPs toxicity on cross-generational earthworms and highlights the hyperactive responses in the antioxidant defense performance of the offsprings. Our findings also underscore the necessity for long-term investigations in assessing the adverse impacts of emerging pollutants.PMID:39552075 | DOI:10.1021/acs.est.4c05208

Signal Transduct Target Ther. 2024 Nov 18;9(1):320. doi: 10.1038/s41392-024-02020-x.ABSTRACTThe heterogeneity of Parkinson's disease (PD) has been recognized in clinical, with patients categorized into distinct subsets based on motor phenotype, such as tremor-dominant PD (TD), postural instability and gait difficulty-dominant PD (PIGD) and mixed PD (Mix). Despite this categorization, the underlying mechanisms of this heterogeneity remain poorly understood, and there is no personalized effective treatment for each PD subtype. To address this, a rat model for PD subtypes was established by unilateral stereotaxic injection of 6-OHDA, followed by cluster analysis of behavioral data. The serum neurofilament light chain (NfL) and uric acid (UA) levels as well as alterations in brain autonomic activity in rats were consistent with clinical patients, and metabolomics results showed that more than 70% of the metabolites in the serum of different subtypes of PD rats and clinical patients appeared to be consistently altered. Further transcriptomic analysis by RNA-seq has elucidated that the development of PD subtypes is associated with altered gene expression in neurotransmitter, neuronal damage in the central or peripheral nervous system, and lipid metabolism. In addition, based on the subtype-specific differentially expressed genes, 25 potential drug candidates were identified. Notably, the Alox15 inhibitor baicalein showed a greater efficacy on Mix rats, highlighting the possibility of selecting targeted treatments for well-defined individuals.PMID:39551798 | DOI:10.1038/s41392-024-02020-x

Environ Res. 2024 Nov 15:120373. doi: 10.1016/j.envres.2024.120373. Online ahead of print.ABSTRACTThe relationship between structure and function in microbial communities is intriguing and complex. In this study, we used single-carbon source domestication to derive consortium YL from the straw-degrading consortium Y. Y and YL exhibited similar straw degradation capabilities, yet YL harbored only half the species diversity of Y, with distinct dominant species. The most enriched microorganisms in Y were Ureibacillus, Acetanaerobacterium, and Hungateiclostridiaceae, whereas Bacillaceae, Bacillus, and Peptostreptococcales-Tissierellales were most enriched in YL. In-depth analysis revealed that Y and YL had comparable abundances of core lignocellulose-degrading genes, as validated by lignocellulolytic enzyme activity assays. However, the number of species harboring these key lignocellulose-degrading genes (K01179, K01181, K00432) in YL was reduced by over 50%, suggesting that functional redundancy enabled YL to maintain similar degradation capabilities to Y despite reduced diversity. Further analyses of key degradative species and co-occurrence networks highlighted the critical functional roles of dominant degradative species within these communities. An analysis of the overall functional pathways in the two microbial consortia revealed distinct metabolic characteristics between them. Pathways such as polycyclic aromatic hydrocarbon degradation and fluorobenzoate degradation were down-regulated in YL compared to Y, a finding corroborated by the metabolomic data. These results suggest a coupling between community structure and functional capacities within these microbial consortia. Overall, our findings deepen our understanding of the structure-function relationship in microbial communities and provide valuable insights for the design of lignocellulose-degrading consortia.PMID:39551375 | DOI:10.1016/j.envres.2024.120373

Life Sci. 2024 Nov 15:123244. doi: 10.1016/j.lfs.2024.123244. Online ahead of print.ABSTRACTAIMS: In light of the evidence supporting a significant role of the gut microbiome in Gulf War Illness (GWI) pathology, we sought to examine its contribution to GWI susceptibility in a mouse model. We also aimed to identify bacterial taxa and microbially-derived metabolites associated with disease susceptibility.MAIN METHODS: Male mice receiving pyridostigmine bromide (PB) orally, and controls were evaluated for symptoms of GWI at 8 weeks post-treatment. The Kansas criteria were adapted to assess behaviors associated with the following domains: gastrointestinal alterations, pain, mood, cognitive function, skin and respiratory disturbances. PB-treated subjects were classified into susceptible (GWI-S) or resilient (GWI-R). The status of the gut microbiome was assessed via analyses of the 16S rRNA gene and microbial-derived metabolites were evaluated with metabolomics tools.KEY FINDINGS: Our results indicate that application of the Kansas criteria to behavioral outcomes in PB-treated mice resulted in a GWI susceptibility rate of ~35 %, similar to the one reported in humans. The composition and structure of the gut microbiome was different in GWI-S subjects compared to both control and GWI-R mice at 8 weeks but differences in microbial community structure were observed prior to PB treatment between GWI-R and GWI-S mice. GWI-S subjects exhibited a pattern of differentially abundant bacterial taxa and microbial metabolites.SIGNIFICANCE: To our knowledge, this is the first preclinical report in which a stratification by susceptibility to GWI and its association with the gut microbiome is described. In light of the research conundrum that vulnerability to GWI represents, the use of tools that could provide further insight into this complex factor should be considered.PMID:39551360 | DOI:10.1016/j.lfs.2024.123244

J Nutr. 2024 Nov 15:S0022-3166(24)01180-5. doi: 10.1016/j.tjnut.2024.11.013. Online ahead of print.ABSTRACTBACKGROUND: The pro-fertility diet is a dietary pattern composed of nutrients and foods most consistently associated with in vitro fertilization (IVF) endpoints in women.OBJECTIVE: We examined the potential biological mechanisms underlying the association between adherence to a pro-fertility diet and IVF endpoints using high-resolution metabolomics.METHODS: Among 120 women undergoing an autologous oocyte IVF cycle (2007-2015) in Northeast United States, we collected a serum sample during controlled ovarian stimulation and a follicular fluid sample on the day of oocyte retrieval. Women completed a food frequency questionnaire upon enrollment into the study to examine adherence to the pro-fertility diet pattern. Liquid chromatography with high-resolution mass spectrometry was used for untargeted metabolomics analysis of biospecimens. We identified metabolic features (and enriched biological pathways) associated with the pro-fertility diet and two IVF endpoints, live birth and clinical pregnancy, via a meet-in-the-middle approach.RESULTS: In the follicular fluid metabolome, vitamin D3 metabolism was associated with adherence to the pro-fertility diet pattern and live birth. Additionally, vitamin D3 metabolism, vitamin B6 metabolism, and bile acid biosynthesis, were associated with both adherence to the pro-fertility diet pattern and clinical pregnancy. In the serum metabolome, only tryptophan metabolism was associated with adherence to the pro-fertility diet pattern and live birth. We confirmed the chemical identity of one metabolite with Level-1 evidence, 4-pyridoxate, which was higher in the serum and follicular fluid among women with stronger adherence to the pro-fertility diet pattern and among women with a live birth.CONCLUSIONS: The beneficial association between adherence to the pro-fertility diet and IVF outcomes may be mediated through vitamin D3 metabolism, vitamin B6 metabolism, and bile acid biosynthesis in the follicular fluid and tryptophan metabolism in the serum. These results provide new insight in the important biological pathways underlying a dietary pattern providing optimal fertility benefits to women.PMID:39551358 | DOI:10.1016/j.tjnut.2024.11.013

Am J Clin Nutr. 2024 Nov 15:S0002-9165(24)00882-7. doi: 10.1016/j.ajcnut.2024.11.011. Online ahead of print.ABSTRACTBACKGROUND: The infant gut microbiome undergoes rapid changes in the first year of life, supporting normal development and long-term health. While diet shapes this process, the role of fibers in complementary foods on gut microbiome maturation is poorly understood.OBJECTIVES: We explored how the transition from human milk to fibers in complementary foods shapes the taxonomic and functional maturation of the gut microbiome within the first year of life.METHODS: We assessed the longitudinal and cross-sectional development of infant gut microbiomes (N=68 infants) and metabolomes (N=33 infants) using linear mixed models to uncover their associations to dietary fibers and their food sources. Fiber intakes were assessed with 3-day food records (months 3,6,9,12) relying on CODEX-compliant fiber fraction values, and questionnaires tracked the overall complementary food introduction. Bacterial species were identified and quantified via MetaPhlAn2 from metagenomic data, and metabolomic profiles were obtained using four LC-MS methods.RESULTS: We identified 176 complementary food fiber-bacterial species associations. First plant-based fibers associated to microbiota compositions similar to breastfeeding, and further associated with aromatic amino acid metabolites, including 5-hydroxyindoleacetic acid (Total dietary fiber - Complementary foods (g) - β=3.50, CI 2.48-4.52, p=6.53x10-5). Distinct fibers from different food categories showed unique associations with specific bacterial taxa. Key species such as Faecalibacterium prausnitznii, associated with oat fibers (g/MJ, β=2.18, CI 1.36-2.84, p=6.12x10-6), reflective of maturing microbial communities. Fiber intake during weaning associated with shifts in metabolite profiles, including immunomodulatory metabolites, with fiber effects observed in a source- and timing-dependent manner, implicated in gradual microbiome diversification.CONCLUSIONS: Introducing complementary dietary fibers during the weaning period supports gut microbiome diversification and stabilization. Even minor dietary variations showed significant associations with microbial taxa and functions from the onset of weaning, highlighting the importance of infant dietary recommendations that support the gut microbiome maturation during early life.CLINICAL TRIAL REGISTRATION NUMBER: Clinicaltrials.gov Identifier: NCT01735123. Date of registration: November 24, 2012. https://www.CLINICALTRIALS: gov/ct2/show/NCT01735123.PMID:39551356 | DOI:10.1016/j.ajcnut.2024.11.011

J Ethnopharmacol. 2024 Nov 15:119112. doi: 10.1016/j.jep.2024.119112. Online ahead of print.ABSTRACTETHNOPHARMACOLOGICAL RELEVANCE: Epilepsy (EP) is one of the most prevalent chronic neurological disorders in children, characterised by a prolonged course and a propensity for recurrence. Banxia Baizhu Tianma Decoction (BBTD), a traditional Chinese medicine formula, is commonly employed in the clinical management of EP and has demonstrated satisfactory therapeutic effects.AIM OF THE STUDY: This study aimed to evaluate the anti-epileptic effects of BBTD and to explore its molecular mechanisms.MATERIALS AND METHODS: EP rat model was induced by pentylenetetrazol (PTZ) and treated with BBTD. Parameters such as seizure grade and duration were recorded to evaluate the improvement of BBTD on epileptic behavior. Nissl staining was used to observe the pathological changes in the cerebral motor cortex. The expression levels of the Bax and Bcl-2 in the motor cortex were measured by western blot analysis to assess neuronal damage and apoptosis. The therapeutic action of BBTD was evaluated by examining the levels of neurotransmitters γ-aminobutyric acid (GABA) and glutamate (Glu) in the brain tissue of EP rats, along with assessments of neuronal damage and apoptosis. Non-targeted metabolomics techniques were employed to conduct a comprehensive analysis of serum and urine metabolites, and network analysis of metabolite-related targets was performed to enhance understanding of the anti-epileptic effects and mechanisms of BBTD.RESULTS: After BBTD treatment, the EP model rats exhibited reduced seizure severity and shortened seizure duration. Moreover, BBTD mitigated PTZ-induced neuronal damage, as evidenced by a significant increase in the number of Nissl bodies in the motor cortex following treatment. At the same time, BBTD inhibited neuronal apoptosis, as demonstrated by the up-regulation of the anti-apoptotic protein Bcl-2 and down-regulation of the pro-apoptotic protein Bax in the brain tissue of treated rats. In addition, BBTD reversed the decreased levels of GABA and the increased levels of Glu in the brain tissue of the model group. Metabolomics analyses suggested that BBTD treatment for EP may be closely associated with alterations in urinary metabolites related to vitamin B6 and pyrimidine metabolism, as well as serum metabolites involved in purine metabolism, glycerophospholipid metabolism and vitamin B6 metabolism. Finally, network analysis of metabolite targets indicated that dopamine and alpha-linolenic acid metabolites may play significant roles in the therapeutic effects of BBTD on EP.CONCLUSION: BBTD demonstrated anti-epileptic effects in PTZ-induced seizure rats by regulating neurotransmitter balance, reducing neuronal damage and inhibiting apoptosis, suggesting its potential for the development of novel AEDs. This is the first time that UHPLC-MS-based urine and serum metabolomics have been used to elucidate the anti-epileptic mechanism of BBTD, providing insights into the underlying mechanisms of BBTD's action.PMID:39551285 | DOI:10.1016/j.jep.2024.119112

Microbes Infect. 2024 Nov 15:105438. doi: 10.1016/j.micinf.2024.105438. Online ahead of print.ABSTRACTThe fungal disease chytridiomycosis (causative agent Batrachochytrium dendrobatidis [Bd]) is a primary contributor to amphibian species declines. The morphological and physiological reorganisation that occurs during amphibian metamorphosis likely increases the vulnerability of metamorphs to Bd. To address this, we exposed pro-metamorphic tadpoles of Fleay's barred frog (Mixophyes fleayi) to Bd and sampled skin and liver sections from control and exposed animals throughout metamorphosis (Gosner stages 40, 42 and 45). We used an untargeted metabolomics approach to assess the metabolic impacts of Bd infection during the critical metamorphic stages, extracting metabolites from sampled tissues and analysing them via Nuclear Magnetic Resonance spectrometry. Most exposed animals became moribund at Gosner stage 45, while a subset seemingly cleared their infections. Metabolite abundance varied throughout development, with Gosner stage 45 samples distinct from previous stages. Clinically infected animals at Gosner stage 45 exhibited profound metabolic dysregulation (e.g., upregulation of amino acid biosynthesis and degradation) in comparison to uninfected groups (negative controls and 'cleared' animals). Despite showing parallels with previous metabolomic analyses of Bd-infected adult frogs, we identified variations in our results that could be attributed to the dramatic changes that characterise metamorphosis and may be driving the heightened vulnerability observed in metamorphic amphibians.PMID:39551241 | DOI:10.1016/j.micinf.2024.105438

J Dairy Sci. 2024 Nov 15:S0022-0302(24)01298-0. doi: 10.3168/jds.2024-25632. Online ahead of print.ABSTRACTLacticaseibacillus rhamnosus Probio-M9 (Probio-M9), a probiotic strain sourced from healthy breast milk, is recognized for its resilience to gastric and bile acids, along with its potential health benefits for infants. Its unique origin may influence its metabolic properties and effectiveness, garnering increasing interest within the scientific community in recent years. However, research on its efficacy as a starter culture for milk fermentation and the associated metabolic shifts remains limited. This study aimed to track the dynamic metabolomic changes of Probio-M9 during the fermentation process. We utilized Probio-M9 as the sole inoculant for milk fermentation, collecting samples at 1.5 to 3-h intervals to monitor the fermentation progression. Moreover, the metabolomics changes of the fermented milk were investigated after a 28-d storage period to evaluate post-storage stability. Metabolite profiles were generated using ultra-high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MSE). A total of 34 representative differential metabolites were identified, primarily comprising peptides and saccharides, followed by acids, amino acids, alcohols, aldehydes, ketones, and intermediate metabolites. Notably, substantial alterations in metabolite levels were observed between 10.5 to 12 h into the fermentation process. Functional metabolites like syringaldehyde, leucic acid, and gentianose accumulated toward the end of fermentation, while peptides emerged as the main differential metabolites after the 28-d storage period. The study offers novel insights into the metabolic dynamics of Probio-M9 during fermentation and storage, which may inform the optimization of Probio-M9 fermented milk production processes. However, it is important to note that the focus of this study on a single starter strain may limit the generalizability of these findings.PMID:39551178 | DOI:10.3168/jds.2024-25632

Ecotoxicol Environ Saf. 2024 Nov 16;288:117348. doi: 10.1016/j.ecoenv.2024.117348. Online ahead of print.ABSTRACTCyanobacterial blooms and aquatic macrophytes can affect the health, physiology, and behavior of freshwater fish. Changes in food intake can be a key indicator of stress in teleost fish, while changes in metabolite abundance in the gut can indicate a shift in metabolic priorities, including response to environmental stressors. Here, we exposed stone moroko (Pseudorasbora parva) to the cyanobacterium Microcystis aeruginosa and/or the macrophyte Ottelia acuminata and analyzed changes in fish health, appetite regulation, and intestinal metabolome after 96-h exposures. We found that O. acuminata treatment didn't change the tested indicators, while exposure to M. aeruginosa increased concentrations of appetite-inhibiting factors, such as CART and GLP-1, and decreased concentrations of stimulatory factors like orexin. Exploration of the metabolome following exposure revealed that the appetite-inhibiting influence of M. aeruginosa was positively correlated with key metabolites of lipid, amino acid, and cholesterol metabolism, especially those associated with bile acid synthesis and secretion. Further, the presence of O. acuminata decreased the adverse effects of M. aeruginosa among neuro-endocrine regulatory factors, which could be explained by altered regulation of intestinal amino acid metabolites. The deeper mechanism by which O. acuminata moderates the harmful effects of M. aeruginosa remains to be identified.PMID:39550875 | DOI:10.1016/j.ecoenv.2024.117348

Talanta. 2024 Nov 12;284:127211. doi: 10.1016/j.talanta.2024.127211. Online ahead of print.ABSTRACTCarotid artery stenosis is mainly produced due to the progressive accumulation of atherosclerotic plaque in the vascular wall. The atherosclerotic plaque is characterized by the accumulation of lipids, low density proteins, expression of chemokines and adhesion molecules, and migration of monocytes and lymphocytes into the plaque. Its rupture can produce stroke, but embolic propensity depends principally on the composition and vulnerability of plaque rather than the severity of stenosis. It is important, then, to ascertain which patients with carotid artery stenosis have a greater risk of developing neurological symptomatology. Here, we present a metabolomic study by using nuclear magnetic resonance (NMR) spectroscopy in atheroma plaque and serum samples from patients with recently symptomatic and asymptomatic carotid stenosis to search for metabolites that could be used as biomarkers associated with plaque vulnerability and subsequent risk of rupture. Thirty-eight atheromatous plaque samples (24 asymptomatic patients and 14 symptomatic) and 70 serum samples (43 asymptomatic and 27 symptomatic) were studied by NMR spectroscopy. The data were analysed using multivariate statistics (PLS-DA) to determine a model to discriminate between symptomatic and asymptomatic samples (atheroma plaques and sera). The calculated PLS-DA models showed a 100 % sensitivity and a 96.6 % specificity for the cross validation to discriminate between symptomatic and asymptomatic plaques, and 88.37 % sensitivity and 77.78 % specificity when serum samples were analysed. According to the results of our multivariate and univariate analysis, the most discriminative metabolites for plaque vulnerability were threonine in serum samples, and glutamate in plaque samples. Also, an analysis of the main metabolic pathways involved in plaque vulnerability revealed that d-glutamine and d-glutamate metabolism, and phenylalanine, tyrosine, and tryptophan biosynthesis were the most affected pathways in plaque and serum, respectively.PMID:39550810 | DOI:10.1016/j.talanta.2024.127211

Sci Rep. 2024 Nov 16;14(1):28270. doi: 10.1038/s41598-024-79481-1.ABSTRACTThe understanding of the role played by extracellular vesicles (EVs) in different tissues has improved significantly in the last years, but remains limited concerning the conjunctiva, a complex eye tissue whose role is pivotal for corneal protection. Here, we conducted a comparative study to isolate and characterize EVs from human conjunctival epithelial (IM-HConEpiC) and human conjunctival mesenchymal stromal cell (Conj-MSCs) secretomes using different isolation methods: differential ultracentrifugation (UC), and a combination of ultrafiltration (UF) with precipitation or size exclusion chromatography (SEC). EVs were characterized by total protein content, size, morphology, and expression of protein markers. EV functional effect was tested in an in vitro oxidative stress model. We successfully recovered EVs with the three methods, although significantly higher yields were obtained with UF-precipitation. Dynamic light scattering analysis confirmed the presence of nano-sized particles, being UC-isolated EVs larger than those isolated by UF-precipitation and UF-SEC. Atomic Force Microscopy showed EVs with a slightly ellipsoidal morphology. EVs enriched with UF-precipitation method were further analyzed, confirming the expression of Alix, CD63, TSG101, and Syntenin-1 by Western blotting and showing that Conj-MSC-derived EVs significantly reduced oxidative stress on IM-HConEpiC. Therefore, we conclude that UF-precipitation is the most efficient method for conjunctival EV enrichment.PMID:39550477 | DOI:10.1038/s41598-024-79481-1

Cancer Cell Int. 2024 Nov 16;24(1):381. doi: 10.1186/s12935-024-03573-1.ABSTRACTBACKGROUND: Despite advancements in therapeutic approaches, including taxane-based chemotherapy and androgen receptor-targeting agents, metastatic castration-resistant prostate cancer (mCRPC) remains an incurable tumor, highlighting the need for novel strategies that can target the complexities of this disease and bypass the development of drug resistance mechanisms. We previously demonstrated the synergistic antitumor interaction of valproic acid (VPA), an antiepileptic agent with histone deacetylase inhibitory activity, with the lipid-lowering drug simvastatin (SIM). This combination sensitizes mCRPC cells to docetaxel treatment both in vitro and in vivo by targeting the cancer stem cell compartment via mevalonate pathway/YAP axis modulation.METHODS: Here, using a combined proteomic and metabolomic/lipidomic approach, we characterized tumor samples derived from 22Rv1 mCRPC cell-xenografted mice treated with or without VPA/SIM and performed an in-depth bioinformatics analysis.RESULTS: We confirmed the specific impact of VPA/SIM on the Hippo-YAP signaling pathway, which is functionally related to the modulation of cancer-related extracellular matrix biology and metabolic reprogramming, providing further insights into the molecular mechanism of the antitumor effects of VPA/SIM.CONCLUSIONS: In this study, we present an in-depth exploration of the potential to repurpose two generic, safe drugs for mCRPC treatment, valproic acid (VPA) and simvastatin (SIM), which already show antitumor efficacy in combination, primarily affecting the cancer stem cell compartment via MVP/YAP axis modulation. Bioinformatics analysis of the LC‒MS/MS and 1H‒NMR metabolomics/lipidomics results confirmed the specific impact of VPA/SIM on Hippo-YAP.PMID:39550583 | DOI:10.1186/s12935-024-03573-1

Virol J. 2024 Nov 16;21(1):295. doi: 10.1186/s12985-024-02571-z.ABSTRACTMounting evidence suggests that the gut-heart axis is critical in the pathogenesis of cardiovascular diseases. The gut serves as the primary pathway through which Coxsackievirus B3 (CVB3) infects its host, leading to acute viral myocarditis (AVMC). However, little is known about the role of gut microflora and its metabolites in the development of AVMC. The AVMC model was established by intraperitoneal injection of CVB3 in mice. Then, 16S ribosomal RNA (16S rRNA) gene sequencing and ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) untargeted metabolomics profiling were performed to analyze the microflora composition and metabolic profile of colonic contents. Compared to the Control mice, the AVMC mice displayed a significant reduction in gut microflora richness and diversity, as revealed by an increased abundance of Proteobacteria and a decreased abundance of Cyanobacteria and Desulfobacterota. LEfSe analysis indicated that the main genera differing between the two groups were Escherichia-Shigella, Lactobacillus, Clostridium_sensu_stricto_1, Prevotellaceae_UCG-001, and Odoribacter. Based on the criterion of OPLS-DA VIP ≥ 1.0 and p-value < 0.05, a total of 198 differential metabolites (DMs) were identified in the gut, including 79 upregulated and 119 downregulated metabolites, of which lipids and lipid-like molecules accounted for the largest proportion. Notably, both altered gut bacterial taxa and metabolites were significantly enriched in the Lipid metabolism pathway, with Traumatic acid (TA), Alpha-Linolenic acid (ALA), Eicosapentaenoic acid (EPA), and Docosahexaenoic acid (DHA) being the key DMs in the pathway. Additionally, significant positive correlations (|r| > 0.80 and p < 0.05) were found between TA levels and Anaerotruncus and Bilophila abundance, between EPA levels and Clostridium_sensu_stricto_1 abundance, and between DHA levels and Escherichia-Shigella abundance, respectively. CVB3 infection leads to notable alterations in gut microflora composition and its metabolic profile, which may participate in AVMC development. Our findings provide important clues for future in-depth studies on AVMC etiology.PMID:39550578 | DOI:10.1186/s12985-024-02571-z

BMC Cardiovasc Disord. 2024 Nov 16;24(1):654. doi: 10.1186/s12872-024-04306-y.ABSTRACTBACKGROUND: The spontaneous echo contrast (SEC) in patients with atrial fibrillation (AF) indicates a prethrombic state that ultimately progresses into thrombus formation. A comprehensive understanding of specific plasma metabolomics characteristics may protect AF patients from thrombus, particularly in the early stage.OBJECTIVES: Through the investigation of metabolic pathways, we endeavor to uncover the metabolomic characteristics associated with SEC states, and to examine the differential metabolites by which may exert their influence on thrombotic states.METHODS: Patients with AF were enrolled, and the participants were divided into three groups based on the results of the echocardiogram: non-SEC, low-SEC and high-SEC group. Samples were collected and subjected to non-targeted metabolomics analysis. The analytical process included data quality control, metabolite difference analysis, component analysis, Kegg cluster analysis, etc. RESULTS: Our metabolic phenotype revealed a clear differential metabolic pattern between the SEC and non-SEC. Specifically, we identified 35 and 142 significantly differential metabolites in venous and atrial plasma, respectively, suggesting that SEC may be involved in pervasive metabolic dysregulation and that the degree of metabolic dysregulation in atrial plasma is more severe than that in venous blood.CONCLUSION: Patients with SEC have a significantly different metabolic pattern compared to those without SEC. Our work promoted the understanding of mechanism of the occurrence and development of SEC, facilitated the screening of the target metabolites for its therapeutic intervention, and provided evidence for the prevention and treatment of SEC or thrombosis in AF. Our work also provided new directions for subsequent research in related fields. In conclusion, our study not only provides a theoretical basis for understanding the occurrence and development of SEC in AF, but also provides recommendations for the daily diet of AF patients with SEC, such as a balanced intake of essential amino acids, avoiding excessive intake of benzoic acid, and intake of appropriate inositol.CLINICAL TRIAL NUMBER: Not applicable.PMID:39550544 | DOI:10.1186/s12872-024-04306-y