PubMed

Rapid Commun Mass Spectrom. 2025 Jul 15;39(13):e10045. doi: 10.1002/rcm.10045.ABSTRACTSingle-cell metabolomics is an emerging and powerful technology that uncovers intercellular heterogeneity and reveals microenvironmental dynamics in both physiological and pathological conditions. This technology enables detailed observations of cellular interactions, providing valuable insights into processes such as aging, immune responses, and disease development. Despite significant advances, the need for detailed discussions on sampling and analytical methods in single-cell metabolomics continues to grow, with increasing focus on selecting the most suitable techniques for diverse research objectives. This review addresses these challenges by exploring key sampling and analytical strategies used in single-cell metabolomics. We focus on three main approaches: the capture and isolation of specific cell types, the precise aspiration of individual cells, and in situ mass spectrometry imaging. These methods are critically assessed to highlight strategies for achieving accurate metabolite detection at the single-cell level across diverse research applications.PMID:40223194 | DOI:10.1002/rcm.10045

Gut Microbes. 2025 Dec;17(1):2486515. doi: 10.1080/19490976.2025.2486515. Epub 2025 Apr 13.ABSTRACTLiver injury is an independent risk factor for multiple organ dysfunction and high mortality in patients with sepsis. However, the pathological mechanisms and therapeutic strategies for sepsis-associated liver injury have not been fully elucidated. Time-restricted feeding (TRF) is a promising dietary regime, but its role in septic liver injury remains unknown. Using 16S rRNA gene sequencing, Q200 targeted metabolomics, transcriptomics, germ-free mice, Hmgcs2/Lpin1 gene knockout mice, and Aml12 cells experiments, we revealed that TRF can mitigate septic liver injury by modulating the gut microbiota, particularly by increasing Lactobacillus murinus (L. murinus) abundance, which was significantly reduced in septic mice. Further study revealed that live L. murinus could markedly elevate serum levels of metabolite 3-hydroxybutyrate (3-HB) and alleviate sepsis-related injury, while the knockout of the key enzyme for 3-HB synthesis (3-hydroxy-3-methylglutaryl-CoA synthase 2, Hmgcs2) in the liver negated this protective effect. Additionally, serum 3-HB levels were significantly positively correlated with L. murinus abundance and negatively correlated with liver injury indicators in septic patients, demonstrating a strong predictive value for septic liver injury (AUC = 0.8429). Mechanistically, 3-HB significantly inhibited hepatocyte ferroptosis by activating the PI3K/AKT/mTOR/LPIN1 pathway, reducing ACSL4, MDA, LPO, and Fe2+ levels. This study demonstrates that TRF reduces septic liver injury by modulating gut microbiota to increase L. murinus, which elevates 3-HB to activate PI3K/AKT/mTOR/LPIN1 and inhibit hepatocyte ferroptosis. Overall, this study elucidates the protective mechanism of TRF against septic liver injury and identifies 3-HB as a potential therapeutic target and predictive biomarker, thereby providing new insights into the clinical management and diagnosis of septic liver injury.PMID:40223164 | DOI:10.1080/19490976.2025.2486515

BMC Med. 2025 Apr 14;23(1):215. doi: 10.1186/s12916-025-04030-0.ABSTRACTBACKGROUND: Although the associations between cord blood lipidome and neonatal birth weight are established, it remains uncertain whether sexual dimorphism in fetal fat accumulation extends to the relationship between cord blood lipid profiles and neonatal abdominal fat compartments. Understanding these relationships could provide insights into early sex-specific differences in lipid metabolism.METHODS: We conducted lipidomics of umbilical cord blood plasma samples (350 (46.6%) girls and 401 (53.4%) boys) from the Growing Up in Singapore Towards healthy Outcomes (GUSTO) birth cohort. Abdominal fat compartments-superficial subcutaneous adipose tissue (sSAT), deep SAT (dSAT), and intra-abdominal adipose tissue (IAT)-were quantified by magnetic resonance imaging within 2 weeks of birth in 239 subjects. Linear regression models were used to assess sex differences in lipid species associated with abdominal fat compartments.RESULTS: Newborn girls had significantly higher superficial and deep subcutaneous adipose tissue volumes compared to boys, whereas intra-abdominal adipose tissue volumes were similar between sexes. In the pooled analysis, cord blood plasma lipids showed distinct associations with different fat depots: 38 lipid species were associated with sSAT, 4 with dSAT, and 38 with IAT. In sex-stratified analyses, 13 lipids were associated with sSAT in girls and 3 in boys, whereas dSAT showed associations with 45 lipids in boys but none in girls. These sex differences were primarily observed in ether-linked phospholipids and ceramides. Notably, no significant associations were observed between lipids and IAT in either sex, suggesting depot-specific sexual dimorphism in early life.CONCLUSIONS: Our study reveals sexual dimorphism in the associations between cord blood lipidome and abdominal adiposity, suggesting depot-specific patterns in adipose tissue development and lipid metabolism in early life.PMID:40223079 | DOI:10.1186/s12916-025-04030-0

Semin Fetal Neonatal Med. 2025 Apr 9:101639. doi: 10.1016/j.siny.2025.101639. Online ahead of print.ABSTRACTThe GI tract-lung-brain axis refers to the communication network linking the gastrointestinal tract, central nervous system, and respiratory system. This axis is particularly significant in preterm neonates because their immune and nervous systems are undergoing rapid development and thus are susceptible to various conditions influenced by GI tract and lung microbiota that are key mediators in this axis. This communication network is connected via neural, hormonal, and immunological regulatory pathways, all of which are pivotal in disease pathogenesis and health. Here we provide a brief introduction to this axis along with interactive mechanisms and perturbations that can affect this system and the roles they play in health and disease.PMID:40222850 | DOI:10.1016/j.siny.2025.101639

Fungal Biol. 2025 May;129(3):101561. doi: 10.1016/j.funbio.2025.101561. Epub 2025 Feb 26.ABSTRACTCordyceps militaris is a widely cultivated mushroom with multiple medicinal properties. However, the emergence of white mildew disease caused by Calcarisporium cordycipiticola has become a serious dilemma, leading to economic losses in its industrial production. The genome of Ca. cordycipiticola possesses more secondary metabolite biosynthetic gene clusters and a smaller number of genes encoding for carbohydrate-active enzyme secretion than other mycoparasites. To uncover those functional metabolites correlated with the infection process, metabolomic profiles between healthy C. militaris fruit bodies (CK) and healthy (HFB) and diseased (DFB) parts of infected C. militaris fruit bodies by Ca. cordicipiticola were compared based on untargeted metabolomic analyses. The function of different metabolites during the pathogen infection and host response processes were further analyzed according to their respective metabolic pathways. Results of key metabolic pathway analyses suggested that a sterigmatocystin-like metabolite functions as one of the virulence factors of white mildew disease on C. militaris, whereas S-adenosyl-L-methionine represents a hub intermediate in both processes of pathogen infection and host response, highlighting the relevance of methyl group turnovers in this battle. More importantly, the detection of toxic metabolites in diseased C. militaris fruiting bodies suggests that this macrofungus contaminated by Ca. cordycipiticola should not be consumed due to the risk that it may contain related instead toxins. This study hypothesizes on the scenario of key metabolic biosynthesis in the battle between Ca. cordycipiticola and C. militaris. Our instead findings not only shed light on the interaction between the pathogen and the host but also provide crucial insights for the development of effective prevention and control strategies in the future.PMID:40222762 | DOI:10.1016/j.funbio.2025.101561

Fungal Biol. 2025 May;129(3):101553. doi: 10.1016/j.funbio.2025.101553. Epub 2025 Feb 28.ABSTRACTThe orchids usually coexist with mycorrhizal fungi during their growth and development. Numerous studies have substantiated the pivotal regulatory role of Tulasnella sp. mycorrhizal fungi in the germination and growth of orchid seeds. However, there remains a dearth of research elucidating the effects and underlying mechanisms of Tulasnella sp. on the growth, development, and metabolite accumulation in Bletilla striata seedlings. In the current study, metabolomics and transcriptomic analysis were used to reveal the key role of the mycorrhizal fungus Tulasnella sp.BJ1 in the growth and accumulation of secondary metabolites in B. striata. The results demonstrated that the application of BJ1 significantly enhanced the growth and development of B. striata seedlings. In September, the plant weight, tuber diameter, and tuber weight in the BJ1 treatment group reached 44.27 ± 6.79 g, 6.13 ± 0.53 cm, and 23.35 ± 3.06 g, respectively, surpassing those in the control group. The polysaccharide content in the BJ1 treatment group and control group peaked in June, reaching 14.91 ± 2.26 % and 14.38 ± 0.25 %, respectively. Total phenol content in both groups decreased in May and June, and the total phenol content in BJ1 treatment group was significantly lower than that in control group. The significant decrease observed in total phenol content during May and June may be attributed to an increase in proportion of polysaccharides promoted by BJ1. The transcriptome results showed that BJ1 upregulated polysaccharide biosynthesis-related genes, such as mannose phosphatase, transferase, mannose 6-phosphate isomerase, hexokinase, fructose kinase, and glucose 6-phosphate isomerase, as well as genes involved in stilbenes biosynthesis, including hydroxycinnamyltransferase and transcinnamate 4-monooxygenase. Metabolomics data indicated that the content of mannose and seven stilbene compounds in the tubers increased significantly after BJ1 treatment. Interestingly, the accumulation of these compounds corresponds to the pathway of upregulated genes. These findings suggest that an upregulation in mannose synthesis may facilitate the accumulation of polysaccharides in B. striata. Therefore, the current study uncovered that the mycorrhizal fungus Tulasnella sp. BJ1 can not only promote the growth and development of B. striata seedlings and increase tuber yield but also promote the accumulation of polysaccharides and stilbenes.PMID:40222760 | DOI:10.1016/j.funbio.2025.101553

J Ethnopharmacol. 2025 Apr 11:119801. doi: 10.1016/j.jep.2025.119801. Online ahead of print.ABSTRACTETHNOPHARMACOLOGICAL RELEVANCE: Schisandra chinensis is used as a traditional Chinese medicine to treat a variety of diseases. Schisandra chinensis lignans (SCL) are one of the most active components extracted from Schisandrae chinensis fructus, exhibit a broad array of pharmacological properties, especially anti-inflammatory and hepatic lipid-lowering effects, suggesting SCL may have potential anti-nonalcoholic steatohepatitis (NASH) ability. However, the therapeutic efficacy of SCL against NASH and the underlying mechanism remains unclear.AIM OF THE STUDY: In the current study, we aimed to investigate the anti-NASH action of SCL and explore the underlying mechanism in vitro and in vivo. We also assess the involvement of the gut-liver axis in the anti-NASH effects of SCL.METHODS: Palmitic acid (PA)-treated HepG2 cells, mouse primary hepatocytes (MPHs) and methionine-choline deficient (MCD) diet-fed mice were selected as NASH models. ORO staining and qRT-PCR were performed to assess hepatic steatosis and inflammatory responses, respectively. Protein expression was detected by Western blotting or immunohistochemistry. The changes of gut microbiota were analyzed using 16S rDNA sequencing in mice. The levels of metabolites in liver and feces were measured by metabolomics.RESULTS: The results showed that SCL treatment alleviated steatosis and inflammation in palmitic acid (PA)-treated HepG2 cells and mouse primary hepatocytes (MPHs). SCL treatment suppressed the phosphorylation of key components involved in NF-κB signaling and enhanced the expression of fatty acid oxidation (FAO)-related enzymes (e.g. CPT1, HMGCS2, and ACOX1) in PA-treated HepG2 cells. SCL could ameliorate hepatic steatosis and inflammation in NASH mice. SCL also ameliorated intestinal barrier injury and restructured the gut microbiota in NASH mice. SCL also modulated hepatic and colonic bile acid metabolism via FXR signaling.CONCLUSION: These findings indicate that SCL treatment ameliorates hepatic inflammation and steatosis in NASH, potentially though to the suppression of NF-κB signaling and the promotion of fatty acid β-oxidation. Moreover, SCL could restore gut microbiota-mediated bile acid homeostasis via activation of FXR/FGF15 signaling. Our study presents a pharmacological rationale for using SCL in the management of NASH.PMID:40222688 | DOI:10.1016/j.jep.2025.119801

J Steroid Biochem Mol Biol. 2025 Apr 11:106760. doi: 10.1016/j.jsbmb.2025.106760. Online ahead of print.ABSTRACTP450 oxidoreductase (POR) facilitates electron flux to type 2 microsomal P450 cytochrome enzymes (CYPs), including the adrenal steroidogenic enzymes CYP17A1 and CYP21A2. Due to the combined impairment of these enzymes, POR deficiency (PORD), an autosomal recessive condition, results in congenital adrenal hyperplasia characterised by combined glucocorticoid and postnatal sex steroid deficiency. This study focuses on urinary steroid excretion in infants affected by PORD in the first week of life. We report on three neonatal PORD cases from two families. One family had two affected babies born three years apart who were stillborn and first-day deceased, respectively. DNA sequencing revealed a homozygous 3bp deletion in exon six leading to an glutamic acid deletion (p.[Glu217del]). Bladder contents were obtained from the stillborn baby, and excreted urine was obtained from the second baby. In a second family, their second affected newborn, antenatally diagnosed carrying the common homozygous p.(Ala287Pro) mutation, had urine collected daily during the first week of life. Steroid excretions were quantified by gas chromatography-mass spectrometry (GC-MS). The birth-day excretions were very similar in all babies. Most notable and unusual was a large excretion of unmetabolised corticosterone, suggesting inhibited catabolism to allow maximum active gluco- and mineralocorticoid availability at birth. Because CYP3A7 (16α-hydroxylase) requires POR, there was an almost complete absence of usually dominant 3β-hydroxy-Δ5 steroids (16α-OH-DHEA and 16α-OH-pregnenolone) and the usually characteristic precursor pregnenolone metabolite 5-pregnene-3β,20α-diol (pregnenediol, 5PD). In the baby sequentially studied over a week, we observed gradual maturation to the typical and familiar PORD neonatal metabolome. At the end of the period, the minimally catabolised corticosterone had diminished, and steroid excretion was completely dominated by 5PD, excreted as both mono- and disulphate conjugates. Whether this metabolome is distinctive of all PORD infants, not just those with severe manifestation, is not known. On the first day of life, standard diagnostic markers are compromised due to fetal-placental-maternal contribution and unique neonatal steroid metabolism. However, the Day 1 PORD steroid metabolome remains distinctive, and we propose using additional biochemical markers reflective of the near complete reduction of POR-dependent CYP3A7 (16α-hydroxylase) activity to improve diagnostic yield.PMID:40222686 | DOI:10.1016/j.jsbmb.2025.106760

J Dairy Sci. 2025 Apr 11:S0022-0302(25)00230-9. doi: 10.3168/jds.2024-25872. Online ahead of print.ABSTRACTExtracellular vesicles (EVs) released by cells contain mRNAs, miRNAs, lncRNAs, lipids, and proteins, playing crucial roles in cell-cell communication. While full-length mRNA transcripts have been documented in EVs secreted by cancer cells, there are no reports on full transcripts secreted by embryos. Our study aimed to identify EV mRNAs in the culture media of bovine embryos and investigate their roles in embryo-maternal communication. Following the isolation of EVs from in vitro fertilization media samples and RNA sequencing, we identified a full mRNA transcript of DPPA3, known to play an essential role in embryo development. To examine the role of DPPA3 in embryo-maternal communication, an in vitro transcribed mRNA of DPPA3 was transfected into bovine endometrial epithelial cells. Transfected and control cells were subsequently analyzed with RNA sequencing and proteomics to assess the effects of DPPA3 on gene expression. A total of 24 genes were found to be upregulated, and one gene was downregulated (FDR <0.01) following DPPA3 transfection, many with known functions in pregnancy recognition. Proteomic analysis revealed 28 differentially expressed proteins, with 19 upregulated and 15 downregulated. Two proteins, ISG15 and MX1, overlapped with the differentially expressed mRNAs. To mimic the natural transfer of EVs from embryos to endometrial cells, we performed coculture with day-8 blastocysts or supplemented the cells with embryo-conditioned culture media. DPPA3 presence was detected in endometrial cells exposed to embryo-conditioned media after just 30 min. Overall, our study highlights the significant role of EVs in cell-cell communication through mRNA signaling from the embryo to the mother.PMID:40222672 | DOI:10.3168/jds.2024-25872

Mucosal Immunol. 2025 Apr 11:S1933-0219(25)00040-6. doi: 10.1016/j.mucimm.2025.04.002. Online ahead of print.ABSTRACTRoux-en-Y gastric bypass (GBP) surgery is an effective treatment for reducing body weight and correcting metabolic dysfunction in individuals with severe obesity. Herein, we characterize the differences between very low energy diet (VLED) and GBP induced weight loss by multi-omic analyses of microbiome and host features in a non-randomized, controlled, single-center study. Eighty-eight participants with severe obesity were recruited into two arms - GBP versus VLED with matching weight loss for 6 weeks and 2-years of follow-up. A dramatic shift in the distribution of gut microbial taxa and their functional capacity was seen in the GBP group at Week 2 after surgery and was sustained through 2 years. Multi-omic analyses were performed after 6 weeks of matching weight loss between the GBP and VLED groups, which pointed to microbiome derived metabolites such as indoxyl sulphate as characterizing the GBP group. We also identified an inverse association between Streptococcus parasanguinis (an oral commensal) and plasma levels of tryptophan and tyrosine. These data have important implications, as they reveal a significant robust restructuring of the microbiome away from a baseline dysbiotic state in the GBP group. Furthermore, multi-omics modelling points to potentially novel mechanistic insights at the intersection of the microbiome and host.PMID:40222615 | DOI:10.1016/j.mucimm.2025.04.002

Clin Chim Acta. 2025 Apr 11:120294. doi: 10.1016/j.cca.2025.120294. Online ahead of print.ABSTRACTBACKGROUND: Trimethylammonium-containing compounds, including choline (CHOL), carnitine (CAR), trimethylglycine (TMG), ergothioneine (ERT), Nε,Nε,Nε-trimethyllysine (TML), γ-butyrobetaine (gBB), and dimethylglycine (DMG) contribute to trimethylamine N-oxide (TMAO) production, a metabolite linked to cardiovascular, renal, and metabolic diseases. An LC-MS/MS method has been established for their simultaneous measurement in human plasma, as an accurate quantification of TMAO and its precursors is crucial for understanding its clinical relevance.METHODS: Blood samples from ten healthy male volunteers were processed using acetonitrile protein precipitation. Analysis was performed using a HILIC column and an isocratic methanol-formic acid mobile phase, achieving a total run time of less than 6 min. Linearity was adequate for all analytes (R2 > 0.995), with mean intra- and inter-assay variation coefficients of 2.88 % and 4.23 %, respectively. Recoveries ranged from 95 % to 101 %, limits of detection from 0.009 to 0.068 µmol/L, and limits of quantification from 0.031 to 0.187 µmol/L. Plasma mean concentrations were 3.18 ± 0.73 µmol/L for TMAO, 3.99 ± 0.65 µmol/L for DMG, 9.84 ± 2.08 µmol/L for CHOL, 24.22 ± 6.19 µmol/L for TMG, 0.54 ± 0.22 µmol/L for gBB, 57.29 ± 8.89 µmol/L for CAR, 1.10 ± 0.42 µmol/L for ERT, and 0.40 ± 0.11 µmol/L for TML. Significant inter-individual variability (mean RSD% of 26 %) was observed.CONCLUSION: The developed LC-MS/MS method enables rapid, sensitive, and selective quantification of TMAO and its precursors in human plasma. The analytical performance supports its application in clinical and metabolomic studies, contributing to a better understanding the role of TMAO in disease states.PMID:40222542 | DOI:10.1016/j.cca.2025.120294

Bioresour Technol. 2025 Apr 11:132515. doi: 10.1016/j.biortech.2025.132515. Online ahead of print.ABSTRACTThis study investigates how high salt stress enhances lipid buildup and phenol removal in Ankistrodesmus sp. ACC. NaCl is a better trigger for treating phenol wastewater (nearly 100 % nitrogen, phosphorus and phenol removal) and producing high-quality biodiesel (0.37 g/L•d). Multi-omics data revealed significant increases in nicotinamide-adenine-dinucleotide phosphate (NADPH), adenosine-triphosphate (ATP), Ca-dependent kinases, serine/threonine-protein kinase, and the Na/Ca exchanger to mitigate Na+-induced ROS. Elevated ATP and NADPH levels support increased activity in the pyruvate-malate cycle and electron transport activity system (ETAS), which enhances phenol removal and further promotes the tricarboxylic acid cycle (TCA). The TCA cycle operates to generate a carbon skeleton to provide energy for microalgal metabolism or growth. In addition, nitrogen pollutants are converted into starch and triglycerides, providing a material energy basis for the adaptation of microalgae to salt stress. These insights deepen our understanding of contaminants degradation and lipid synthesis by microalgae under high salt stress.PMID:40222491 | DOI:10.1016/j.biortech.2025.132515

Poult Sci. 2025 Mar 31;104(6):105111. doi: 10.1016/j.psj.2025.105111. Online ahead of print.ABSTRACTThis study investigated the differences of calcium transport-related proteins and metabolites in the uterus of hens with different breaking strength eggshell during the eggshell calcification. A total of 200 Hy-Line Brown laying hens, aged 75 weeks, were selected and categorized into two groups based on the eggshell breaking strength: a high-strength group (HS, > 42 N) and a low-strength group (LS, < 32 N). Laying hens were sampled at 1 h, 7 h (the initiation stage of eggshell calcification), and 17 h (the growth stage of eggshell calcification) post-oviposition (PO). The LS group showed a decreased thickness, weight and weight ratio of eggshells, accompanied with ultrastructural deterioration and total Ca reduction. The expression levels of ATP2A3, ATP2B2, SLC8A1, and SLC8A3 were significantly increased in the HS at 17 h PO when compared to 1 h and 7 h PO, while no significant changes were observed in the LS. At 7 h PO, the LS group had lower uterine mucosa calcium levels, higher TRPV6 protein expression, and lower CALB1 protein expression. In the HS group, uterine metabolites showed a significant increase in glutathione, citrulline, and proline at 7 h PO, whereas, at 17 h PO, the tricarboxylic acid cycle pathway was significantly enriched. These findings suggest that uterine calcium transport activity is relatively subdued during the initiation stage of eggshell calcification, focusing on redox repair activities to maintain homeostasis for mammillary knobs formation. Subsequently, uterine calcium transport activity becomes highly active during the growth stage of eggshell calcification, primarily supporting rapid calcium transport through enhanced energy metabolism. In aged laying hens, the lower eggshell breaking strength may be attributed to decreased calcium levels during the initiation stage and imbalanced redox during the growth stage, which could affect calcium transport and lead to a weak ultrastructure during the calcification period.PMID:40222347 | DOI:10.1016/j.psj.2025.105111

Biochem Biophys Res Commun. 2025 Apr 5;763:151766. doi: 10.1016/j.bbrc.2025.151766. Online ahead of print.ABSTRACTCancer cachexia is a multifactorial metabolic syndrome characterized by progressive weight loss, muscle wasting, and systemic inflammation. Despite its clinical significance, the underlying mechanisms linking central and peripheral metabolic changes remain incompletely understood. In this study, we employed a murine model of cancer cachexia induced by intraperitoneal injection of Lewis lung carcinoma (LLC1) cells to investigate tissue-specific metabolic adaptations. Cachectic mice exhibited reduced food intake, body weight loss, impaired thermoregulation, and decreased energy expenditure. Metabolomic profiling of serum, skeletal muscle, and hypothalamus revealed distinct metabolic shifts, with increased fatty acid and ketone body utilization and altered amino acid metabolism. Notably, hypothalamic metabolite changes diverged from peripheral tissues, showing decreased neurotransmitter-related metabolites and enhanced lipid-based energy signatures. Gene expression analysis further confirmed upregulation of glycolysis- and lipid oxidation-related genes in both hypothalamus and muscle. These findings highlight coordinated yet compartmentalized metabolic remodeling in cancer cachexia and suggest that hypothalamic adaptations may play a central role in the systemic energy imbalance associated with cachexia progression.PMID:40222332 | DOI:10.1016/j.bbrc.2025.151766

Biomaterials. 2025 Mar 21;321:123283. doi: 10.1016/j.biomaterials.2025.123283. Online ahead of print.ABSTRACTOBJECTIVE: This study investigates the role of Gelatin-Catalase (Gel@CAT)-L hydrogel in mediating reactive oxygen species (ROS) production and maintaining mitochondrial homeostasis through SIRT3-mediated unfolded protein response (UPRmt), while exploring its involvement in the molecular mechanism of osteoarthritis (OA).METHODS: Self-assembled Gel@CAT-L hydrogels were fabricated and characterized using transmission electron microscopy, mechanical testing, external release property evaluation, and oxygen production measurement. Biocompatibility was assessed via live/dead cell staining and CCK8 assays. An OA mouse model was established using destabilization of the medial meniscus (DMM) surgery. X-ray and micro-CT imaging were employed to evaluate the structural integrity of the mouse knee joints, while histological staining was used to assess cartilage degeneration. Immunohistochemistry was performed to analyze the expression of proteins including Col2a1, Aggrecan, MMP13, ADAMTS5, SIRT3, PINK1, and Parkin. Multi-omics analyses-encompassing high-throughput sequencing, proteomics, and metabolomics-were conducted to identify key genes and metabolic pathways targeted by Gel@CAT-L hydrogel intervention in OA. Immunofluorescence techniques were utilized to measure ROS levels, mitochondrial membrane potential, and the expression of SIRT3, PINK1, Parkin, LYSO, LC3B, Col2a1, and MMP13 in primary mouse chondrocytes and mouse knee joints. Flow cytometry was applied to quantify ROS-positive cells. RT-qPCR analysis was conducted to determine mRNA levels of Aggrecan, Col2a1, ADAMTS5, MMP13, SIRT3, mtDNA, HSP60, LONP1, CLPP, and Atf5 in primary mouse chondrocytes, mouse knee joints, and human knee joints. Western blotting was performed to measure protein expression levels of SIRT3, HSP60, LONP1, CLPP, and Atf5 in both primary mouse chondrocytes and mouse knee joints. Additionally, 20 samples each from the control (CON) and OA groups were collected for analysis. Hematoxylin and eosin staining was used to evaluate cartilage degeneration in human knee joints. The Mankin histological scoring system quantified the degree of cartilage degradation, while immunofluorescence analyzed SIRT3 protein expression in human knee joints.RESULTS: In vitro experiments demonstrated that self-assembled Gel@CAT-L hydrogels exhibited excellent biodegradability and oxygen-releasing capabilities, providing a stable three-dimensional environment conducive to cell viability and proliferation while reducing ROS levels. Multi-omics analysis identified SIRT3 as a key regulatory gene in mitigating OA and revealed its central role in the UPRmt pathway. Furthermore, Gel@CAT-L was confirmed to regulate mitochondrial homeostasis. Both in vitro experiments and in vivo mouse model studies confirmed that Gel@CAT-L significantly reduced ROS levels and regulated mitochondrial autophagy by activating the SIRT3-mediated UPRmt pathway, thereby improving the pathological state of OA. Clinical trials indicated downregulation of SIRT3 and UPRmt-related proteins in OA patients.CONCLUSION: Gel@CAT-L hydrogel activates SIRT3-mediated UPRmt to regulate ROS and mitochondrial homeostasis, providing potential therapeutic benefits for OA.PMID:40222260 | DOI:10.1016/j.biomaterials.2025.123283

Food Chem. 2025 Apr 7;483:144251. doi: 10.1016/j.foodchem.2025.144251. Online ahead of print.ABSTRACTKazakh cheese is a traditional fermented dairy product. In this study, high-throughput sequencing, HS-SPME-GC-MS/MS, and untargeted metabolomics were employed to investigate the microbial succession and flavor profiles of Kazakh cheese under traditional handicraft. During processing, Lactobacillus and Acetobacter were the dominant bacterial genera, while Pichia and Kluyveromyces were the predominant yeast genera. The predominant volatile compounds identified across different stages were phenethyl alcohol, acetoin, hexanoic acid, and phenethyl acetate, with their maximum concentrations attained at the cheese during ripening (CR) stage. KEGG pathway enrichment analysis identified amino acid metabolism as the most significantly enriched pathway. Furthermore, Spearman correlation analysis revealed a significant association between Pichia, Lactobacillus, Lactococcus, Kluyveromyces, and flavor compounds, suggesting the crucial role of these microbes in flavor development. This study provides a theoretical foundation for enhancing the quality of traditional fermented Kazakh cheese and advancing Xinjiang's specialty dairy industry.PMID:40222124 | DOI:10.1016/j.foodchem.2025.144251

J Environ Manage. 2025 Apr 12;381:125298. doi: 10.1016/j.jenvman.2025.125298. Online ahead of print.ABSTRACTEnvironmental stressors, such as nanoplastics (NPs) and freeze-thaw cycles (FTC), are increasingly prevalent, posing significant risks to plant health and agricultural productivity. NPs, being persistent and ubiquitous, can disrupt plant physiological processes, while FTC, common in temperate climates, exacerbates the oxidative damage caused by NPs, leading to further impairment of plant cellular structures. This study investigates the combined effects of these stressors on rye seedlings, exposing them to 100 mg/L polystyrene NPs and simulating early winter conditions with temperature fluctuations between 5°C and -5°C. FTC exposure exacerbated oxidative stress, as indicated by increased hydrogen peroxide (H2O2) accumulation and elevated superoxide dismutase (SOD) activity, suggesting severe oxidative damage. Photosynthesis was significantly inhibited, as evidenced by reduced chlorophyll content and net photosynthetic rate (Pn), accompanied by heightened membrane lipid peroxidation, indicating aggravated cellular membrane damage under combined stress conditions. Additionally, metabolomic analysis revealed significant alterations in key metabolic pathways, including the tricarboxylic acid (TCA) cycle, aminoacyl-tRNA synthesis, and lipid metabolism, which were notably influenced by the combined stressors. The activation of the ascorbate-glutathione (AsA-GSH) cycle suggests a protective adaptive response to mitigate oxidative stress. These findings highlight that the interaction between NPs and abiotic stressors, such as FTC, profoundly alters plant physiological and metabolic responses, ultimately compromising plant growth and resilience. This study underscores the necessity of integrated environmental assessments that consider the synergistic effects of multiple stress factors. Such assessments are essential for developing strategies to enhance plant tolerance to escalating environmental pollutants and climate-induced stressors.PMID:40222074 | DOI:10.1016/j.jenvman.2025.125298

J Proteome Res. 2025 Apr 13. doi: 10.1021/acs.jproteome.5c00040. Online ahead of print.ABSTRACTBackground: Metabolic dysfunction-associated steatotic liver disease (MASLD) can be recapitulated in mice fed a high-fat diet. The development of MASLD and the diet per se can both perturb metabolism in key extrahepatic tissues such as the heart, kidney, and skeletal muscle. To date, these alterations have not been well described in this animal model of diet-induced MASLD. Methodology: Male C57BL/6J mice were fed either standard (SC, n = 12) or high-fat chow (HF, n = 11) for 18 weeks. Metabolites were extracted from the heart, kidney, and skeletal muscle and analyzed by 1H nuclear magnetic resonance (NMR) spectroscopy, along with multivariate and univariate statistical analyses. Results: Kidney metabolite profiles exhibited the largest differences between HF and SC diets, followed by those of skeletal muscle and then the heart. Some alterations were common across all tissues, namely decreased trimethylamine and elevated levels of linoleic acid and polyunsaturated fatty acids in HF compared to SC (p < 0.05 for all three metabolites). Overall, the metabolite variations were consistent with shifts in carbohydrate and lipid substrate selection for oxidation, increased tissue stress in the heart and kidneys, and altered choline metabolism. These findings may serve as additional important descriptors of MASLD onset and progression.PMID:40222045 | DOI:10.1021/acs.jproteome.5c00040

Pediatr Allergy Immunol. 2025 Apr;36(4):e70086. doi: 10.1111/pai.70086.ABSTRACTRespiratory infections are a leading cause of morbidity and mortality during the early life period, and experiencing recurrent infections may increase the risk of developing chronic respiratory diseases, such as asthma. Over the last several decades, metabolomics methods have been applied to inform upon the underlying biochemistry of pediatric respiratory infection response, to discriminate between respiratory infection types, and to identify biomarkers of severity and susceptibility. While these studies have demonstrated the power of applying metabolomics to the study of pediatric respiratory infection and contributed to an understanding of respiratory infections during the unique period of immune development, key differences in study design, infection type(s) of interest, biosamples, metabolomics measurement methods, and lack of external validation have limited the translation of these findings into the clinic. The purpose of this review is to summarize overlaps across existing studies of commonly reported metabolomics findings and emphasize areas of opportunity for future study. We highlight several metabolomics pathways-such as the citric acid cycle and sphingolipid metabolism-that have been reported consistently in respiratory infection response. We then discuss putatively identified metabolomic markers to discriminate between respiratory infection types and possible markers of infection severity and proneness. Finally, we close with a summary and perspective of future directions of the field.PMID:40221829 | DOI:10.1111/pai.70086

BMC Biol. 2025 Apr 12;23(1):96. doi: 10.1186/s12915-025-02198-8.ABSTRACTBACKGROUND: Age-related macular degeneration (AMD) is a leading cause of blindness, characterized by retinal pigment epithelium (RPE) dysfunction, extracellular deposit formation, and disrupted lipid metabolism. Understanding the molecular changes underlying AMD is essential for identifying diagnostic markers and therapeutic targets.RESULTS: This multiomic study employed a primary RPE culture model to investigate age-related changes associated with AMD. Over 25 weeks, RPE cells exhibited phenotypic deterioration, including depigmentation, cell shape deformation, and barrier integrity loss, accompanied by extracellular deposit formation. Transcriptomic analysis revealed dysregulation of genes involved in lipid metabolism, including pathways for cholesterol transport, glycerophospholipids, and ceramide biosynthesis. Metabolomic profiling further identified significant changes in glycerophospholipid and sphingolipid metabolism, highlighting a decline in phospholipid species and ceramide accumulation. Serum analysis of AMD patients revealed altered levels of 18 lipids identified in RPE cultures. Four lipids showed significant differences compared to controls: GlcCer(d16:1/18:0) (1.23-fold increase, adj. p value < 0.001), PE(19:1(9Z)/22:2(13Z,16Z)) (0.34-fold decrease, adj. p value < 0.001), PE(15:0/20:3(5Z,8Z,11Z)) (0.66-fold decrease, adj. p value < 0.05), and PC(22:2(13Z,16Z)/13:0) (0.71-fold decrease, adj. p value < 0.05). These findings underscore the systemic nature of lipid dysregulation in AMD and the translational relevance of the RPE model.CONCLUSIONS: This study highlights the significant role of lipid metabolism dysregulation in AMD pathogenesis. The consistent lipidomic alterations observed in RPE cultures and AMD patient serum reinforce their potential as biomarkers for disease progression and therapeutic targets. These findings provide a robust framework for understanding AMD-associated lipid metabolism changes and their systemic impact.PMID:40221802 | DOI:10.1186/s12915-025-02198-8