PubMed

Front Nutr. 2025 Mar 3;12:1522982. doi: 10.3389/fnut.2025.1522982. eCollection 2025.ABSTRACTINTRODUCTION: Hyperlipidemia is regarded as one of the crucial factors leading to atherosclerosis and other cardiovascular diseases. Gut microbiota plays an important role in regulating host lipid metabolism. Nevertheless, the exact mechanisms behind this remain unclear.METHODS: In the present study, a hyperlipidemic zebrafish model was established using a high-cholesterol diet (HCD) to evaluate the anti-hyperlipidemic effects of Lactobacillus fermentum E15 (L. fermentum E15).RESULTS: Results showed that L. fermentum E15 effectively reduced lipid accumulation in the blood vessels and liver of HCD-fed zebrafish larvae. Meanwhile, L. fermentum E15 improved abnormal lipid levels, and normalized liver enzyme activity. Real-time quantitative polymerase chain reaction (RT-qPCR) analysis revealed that L. fermentum E15 downregulated the expression of sterol regulatory element-binding factor (SREBP-1), peroxisome proliferator-activated receptor-gamma (PPAR-γ), and fatty acid synthase (Fasn), while upregulated peroxisome proliferator-activated receptor-alpha (PPAR-α). Additionally, metabolomic analysis revealed that L. fermentum E15 produced a series of short-chain fatty acids (SCFAs), including acetic acid, propionic acid, butyric acid, and isovaleric acid. Notably, isovaleric acid contributed to the reduction of lipid droplet accumulation in the liver and blood vessels of HCD-fed zebrafish larvae. In contrast, blocking G-protein coupled receptor 43 (GPR43) with pertussis toxin (PTX) abolished the effects of L. fermentum E15 and isovaleric acid on reducing lipid accumulation in HCD-fed zebrafish larvae. RT-qPCR results further suggested that both L. fermentum E15 and isovaleric acid promoted the expression of GPR43 and leptin A, which was inhibited by PTX.CONCLUSION: These findings suggested that L. fermentum E15 alleviates HCD-induced hyperlipidemia by activating GPR43 through SCFAs.PMID:40098735 | PMC:PMC11911183 | DOI:10.3389/fnut.2025.1522982

FASEB J. 2025 Mar 31;39(6):e70408. doi: 10.1096/fj.202401669R.ABSTRACTIntestinal microbes can beneficially impact host physiology, prompting investigations into the therapeutic usage of such microbes in a range of diseases. For example, human intestinal microbe Limosilactobacillus reuteri strains ATCC PTA 6475 and DSM 17938 are being considered for use for intestinal ailments, including colic, infection, and inflammation, as well as for non-intestinal ailments, including osteoporosis, wound healing, and autism spectrum disorder. While many of their beneficial properties are attributed to suppressing inflammatory responses, we postulated that L. reuteri may also regulate intestinal hormones to affect physiology within and outside of the gut. To determine if L. reuteri secreted factors impact the secretion of enteric hormones, we treated an engineered jejunal organoid line, NGN3-HIO, which can be induced to be enriched in enteroendocrine cells, with L. reuteri 6475 or 17938 conditioned medium and performed transcriptomics. Our data suggest that these L. reuteri strains affect the transcription of many gut hormones, including vasopressin and luteinizing hormone subunit beta, which have not been previously recognized as produced in the gut epithelium. Moreover, we find that these hormones appear to be produced in enterocytes, in contrast to canonical gut hormones produced in enteroendocrine cells. Finally, we show that L. reuteri conditioned media promote the secretion of enteric hormones, including serotonin, GIP, PYY, vasopressin, and luteinizing hormone subunit beta, and identify by metabolomics metabolites potentially mediating these effects on hormones. These results support L. reuteri affecting host physiology through intestinal hormone secretion, thereby expanding our understanding of the mechanistic actions of this microbe.PMID:40098558 | DOI:10.1096/fj.202401669R

Biopreserv Biobank. 2025 Mar 18. doi: 10.1089/bio.2024.0141. Online ahead of print.ABSTRACTObjectives: Personalized medicine emphasizes prevention and early diagnosis by developing genetic screening and biomarker assessment tools. Biobanks, including University of Piemonte Orientale (UPO) Biobank, support this effort by providing high-quality biological samples collected, processed, and stored using optimized standardized protocols. To determine the optimal long-term storage conditions for biospecimens used in biomedical research, we evaluated plasma and serum samples cryopreserved using two storage methods, cryovials and straws, across various analytical methodologies with differing sensitivity and robustness. Design and Methods: Plasma and serum samples cryopreserved in liquid nitrogen in vials and straw at the UPO Biobank were subjected to multiple analyses including standard biochemical laboratory analysis, targeted lipidomics, untargeted proteomics, and targeted metabolites quantification through mass spectrometry-based analytical techniques. Results: Our data demonstrate the robustness and applicability of both storage methods for standard laboratory analyses in evaluating clinically relevant markers in plasma and serum. Lipidomic analysis revealed slight disparities in lipid abundance, though these differences were mostly confined to specific lipid species, particularly fatty acids. Conversely, proteomic and metabolomic analyses uncovered variations in abundance in a significant, albeit limited, fraction of analytes between vials and straw-derived samples. Conclusions: By highlighting similarities and differences in samples stored in these conditions, this study provides significant insights into optimizing biobanking practices and understanding the factors that influence the integrity of cryopreserved biospecimens and the reliability of the data derived from them. Both straws and vials are convenient and efficient cryopreservation methods, essentially equivalent for samples dedicated to robust and relatively low-sensitive standardized analyses. However, our findings emphasize the need for caution when interpreting omics data from samples subjected to different cryopreservation methods, as subtle variations can arise even with different types of containers.PMID:40098524 | DOI:10.1089/bio.2024.0141

J Agric Food Chem. 2025 Mar 18. doi: 10.1021/acs.jafc.4c12689. Online ahead of print.ABSTRACTConstipation severely affects the quality of life, and current treatment strategies may cause adverse effects. Garlic fructans have shown great potential for improving gastrointestinal health. This study aimed to elucidate the protective effects and mechanisms of three garlic fructans with different degrees of polymerization (DPs) against loperamide (Lop)-induced constipated mice. The results showed that garlic fructans accelerated intestinal motility (0.57-1.05-fold) and alleviated colonic crypt shortening (1.42-1.97-fold). Furthermore, dysregulation of excitatory/inhibitory gastrointestinal hormones was significantly alleviated, accompanied by restored propionic and butyric acid production and ameliorated bile acid metabolism disorders. Garlic fructans also improved gut microbiota homeostasis in constipated mice by inhibiting harmful bacteria and promoting the growth of beneficial bacteria. Compared with high-DP garlic fructans, low-DP fructans exhibited stronger relieving effects on Lop-induced constipation symptoms in mice. In conclusion, this study demonstrated the anticonstipation activity of garlic fructans and revealed that low-DP garlic fructans were more effective in alleviating Lop-induced constipation.PMID:40098447 | DOI:10.1021/acs.jafc.4c12689

Int J Cancer. 2025 Mar 18. doi: 10.1002/ijc.35401. Online ahead of print.ABSTRACTWe assessed hepatocellular carcinoma (HCC) risk associated with smoking and alcohol consumption and their interactions, using both questionnaire data and objective serum biomarkers. Information on smoking and alcohol consumption was collected at baseline from 450,112 participants of the EPIC cohort, among whom 255 developed HCC after a median follow-up of 14 years. In a nested case-control subset of 108 HCC cases and 108 matched controls, known biomarkers of smoking (cotinine, nicotine) and habitual alcohol consumption (2-hydroxy-3-methylbutyric acid) were annotated from untargeted metabolomics features. Multivariable-adjusted hazard ratios (HRs) or odds ratios (ORs) with 95% confidence intervals (CIs) were computed, and multiplicative and additive interaction parameters were calculated. Compared to never smokers, current smokers had a higher HCC risk (HR = 2.46, 95% CI = 1.77-3.43) dose-dependently with the number of cigarettes smoked per day (Ptrend <.001). Compared to light drinkers, HCC risk was higher in former (HR = 3.20, 95% CI = 1.70-6.03), periodically heavy (HR = 1.98, 95% CI = 1.11-3.54), and always heavy (HR = 5.51, 95% CI = 2.39-12.7) drinkers. Higher HCC risk was also observed in the highest versus the lowest tertiles of cotinine (OR = 4.88, 95% CI = 1.52-15.70), nicotine (OR = 5.80, 95% CI = 1.33-25.30) and 2-hydroxy-3-methylbutyric acid (OR = 5.89, 95% CI = 1.33-26.12). Questionnaire-assessed smoking and alcohol exposures did not demonstrate an HCC risk interaction at the multiplicative (MI = 0.88, 95% CI = 0.40-1.96) or additive (RERI = 0.71, 95% CI = -10.1 to 23.6; attributable proportion = 0.17, 95% CI = -0.52 to 1.16; synergy index = 1.27, 95% CI = 0.98-1.66) scales. Similar analyses with cotinine, nicotine, and 2-hydroxy-3-methylbutyric acid also did not show interactions between smoking and alcohol consumption on HCC risk. Smoking and alcohol consumption are strong independent risk factors for HCC and do not appear to synergistically impact its risk, but larger studies are needed.PMID:40098437 | DOI:10.1002/ijc.35401

BMC Ophthalmol. 2025 Mar 17;25(1):137. doi: 10.1186/s12886-025-03961-9.ABSTRACTBACKGROUND: For patients with bilateral age-related cataracts, sequential phacoemulsification and intraocular lens implantation is a common treatment. However, it remains unclear whether surgery on the first eye has an impact on the second eye, as current research results are inconsistent. This study will explore whether surgery on one eye affects the non-operated eye through aqueous humor cytokines and metabolomic analyses in the second eye.METHODS: A rabbit model of unilateral phacoemulsification and intraocular lens implantation was established. The experimental group consisted of 15 rabbits undergoing this procedure. Postoperatively, rabbits were divided into five subgroups (three rabbits per subgroup), and aqueous humor was collected from both the operated and non-operated eyes at 1 day, 3 days, 1 week, 2 weeks, and 3 weeks after surgery. Additionally, 5 rabbits were selected as a control group, from which aqueous humor was extracted. Levels of IL-1a, IL-1β, IL-2, IL-4, IL-6, IL-8, IFN-γ, TNF-α, MCP-1, and VEGF in the aqueous humor were compared. In the clinical study, preoperative aqueous humor samples were collected from 22 patients undergoing bilateral phacoemulsification and intraocular lens implantation. Among them, 11 patients were tested for the aforementioned 10 cytokines, while the other 11 patients underwent untargeted metabolomics research.RESULTS: In the animal experiment, levels of all 10 cytokines in the operated eyes were significantly higher compared to both the control and non-operated eyes groups (P < 0.05). In the non-operated eyes, IL-1β and IL-2 levels were also elevated compared to the control (P < 0.05); however, no statistically significant differences were observed between the non-operated eyes and the control group at postoperative time points of 1 day, 3 days, 1 week, 2 weeks, and 3 weeks. In the clinical study, no significant differences were found in cytokine levels between the two eyes. In the untargeted metabolomics analysis, 354 metabolites showed differential expression, 280 were upregulated and 74 were downregulated. Notably, Adenine and 2-Aminopurine were significantly downregulated, highlighting Purine metabolism as the most impacted pathway.CONCLUSIONS: Animal experiments showed a significant increase in IL-1β and IL-2 levels in the non-operated eyes postoperatively, reflecting systemic and local inflammatory responses. In clinical experiments, although no significant changes in cytokines were observed in the aqueous humor of both eyes, differential expression of metabolites indicated metabolic adjustments in the non-operated eye following surgery on the first eye. These findings reveal that unilateral cataract surgery may affect the stability of the intraocular environment in the contralateral eye, suggesting that in staged bilateral surgeries, potential metabolic changes in the non-operated eye and their clinical significance should be considered. This result provides important reference value for optimizing postoperative management strategies, reducing complications, and determining the timing for bilateral surgeries, warranting further investigation.PMID:40098128 | DOI:10.1186/s12886-025-03961-9

J Ovarian Res. 2025 Mar 17;18(1):56. doi: 10.1186/s13048-025-01625-2.ABSTRACTBACKGROUND: Ovarian aging is the main cause of reduced reproductive life span, yet its metabolic profiles remain poorly understood. This study aimed to reveal the metabolic homogeneity and heterogeneity between physiological and pathological ovarian aging.METHODS: Seventy serum samples from physiological ovarian aging participants, pathological ovarian aging participants (including diminished ovarian reserve (DOR), subclinical premature ovarian insufficiency (scPOI) and premature ovarian insufficiency (POI)), as well as healthy participants were collected and analyzed by untargeted metabolomics.RESULTS: Five homogeneous differential metabolites (neopterin, menaquinone, sphingomyelin (SM) (d14:1/24:2), SM (d14:0/21:1) and SM (d17:0/25:1)) were found in both physiological and pathological ovarian aging. While five distinct metabolites, including phosphoglyceride (PC) (17:0/18:2), PC (18:2e/17:2), SM (d22:1/14:1), SM (d14:1/20:1) and 4-hydroxyretinoic acid were specific to pathological ovarian aging. Functional annotation of differential metabolites suggested that folate biosynthesis, ubiquinone and other terpenoid-quinone biosynthesis pathways, were mainly involved in the ovarian aging process. Meanwhile, dopaminergic synapses pathway was strongly associated with scPOI, vitamin digestion and absorption and retinol metabolism were associated with POI. Furthermore, testosterone sulfate, SM (d14:0/28:1), PC (18:0e/4:0) and 4-hydroxyretinoic acid, were identified as potential biomarkers for diagnosing physiological ovarian aging, DOR, scPOI, and POI, respectively. Additionally, SM (d14:1/24:2) strongly correlated with both physiological and pathological ovarian aging. 4-hydroxyretinoic acid was strongly correlated with pathological ovarian aging.CONCLUSIONS: Metabolic homogeneity of physiological and pathological ovarian aging was related to disorders of lipid, folate, ubiquinone metabolism, while metabolic heterogeneity between them was related to disorders of lipid, vitamin and retinol metabolism.CLINICAL TRIAL NUMBER: Not applicable.PMID:40098062 | DOI:10.1186/s13048-025-01625-2

Infect Dis Poverty. 2025 Mar 18;14(1):21. doi: 10.1186/s40249-025-01291-y.ABSTRACTBACKGROUND: The association between gut microbial metabolites and immunologic non-response among people living with HIV (PLHIV) receiving antiretroviral therapy (ART) has not been well established. We aimed to characterize gut microbial metabolites among HIV-infected men who have sex with men (MSM) with different immunologic responses.METHODS: We recruited HIV-infected MSM from Guangzhou Eighth People's Hospital and HIV-uninfected MSM (healthy controls, HC) from a local MSM community-based organization in Guangzhou between June and October 2021. HIV-infected MSM were grouped into good immunological responders (GIR) (CD4 + T cell count ≥ 350 cells/μl) and poor immunological responders (PIR) (CD4 + T cell count < 350 cells/μl) after 24 months of ART treatment. Online questionnaires and stool samples were collected. Microbial metabolites in stool were obtained through ultra-performance liquid chromatography coupled to a tandem mass spectrometry (UPLC-MS/MS) system. Differential metabolites were identified and analyzed using the Kruskal-Wallis test, followed by pairwise comparisons with the Wilcoxon rank-sum test. The least absolute selection and shrinkage operator was used to select potential metabolites biomarkers.RESULTS: A total of 51 HC, 56 GIR, and 42 PIR were included. No statistically significant differences were observed in the median time since HIV diagnosis and ART duration between GIR and PIR. Among the 174 quantified metabolites, 81 significantly differed among HC, GIR, and PIR (P < 0.05). Among differential metabolites, indole-3-propionic acid significantly decreased from HC (11.39 nmol/g) and GIR (8.16 nmol/g) to PIR (6.50 nmol/g). The pathway analysis showed that tryptophan metabolism differed significantly between GIR and PIR (P < 0.05). Four potential metabolites biomarkers (dimethylglycine, cinnamic acid, 3-hydroxyisovaleric acid, and propionic acid) that distinguish GIR and PIR were identified, and the corresponding area under the curve based on potential biomarkers was 0.773 (95% CI: 0.675-0.871).CONCLUSIONS: This study identified significant differences in gut microbial metabolites among HIV-infected MSM with different immunologic responses. These results indicate the potential of gut microbial metabolites as novel disease progression markers and therapeutic targets.PMID:40098016 | DOI:10.1186/s40249-025-01291-y

Genome Biol. 2025 Mar 17;26(1):59. doi: 10.1186/s13059-025-03531-8.ABSTRACTBACKGROUND: Dyslipidemia or hypercholesterolemia are among the main risk factors for cardiovascular diseases. Unraveling the molecular basis of lipid or cholesterol homeostasis would help to identify novel drug targets and develop effective therapeutics.RESULTS: Here, we adopt a systematic approach to catalog the genes underlying lipid and cholesterol homeostasis by combinatorial use of high-throughput CRISPR screening, RNA sequencing, human genetic variant association analysis, and proteomic and metabolomic profiling. Such integrative multi-omics efforts identify gamma-glutamyltransferase GGT7 as an intriguing potential cholesterol and lipid regulator. As a SREBP2-dependent target, GGT7 positively regulates cellular cholesterol levels and affects the expression of several cholesterol metabolism genes. Furthermore, GGT7 interacts with actin-dependent motor protein MYH10 to control low-density lipoprotein cholesterol (LDL-C) uptake into the cells. Genetic ablation of Ggt7 in mice leads to reduced serum cholesterol levels, supporting an in vivo role of Ggt7 during cholesterol homeostasis.CONCLUSIONS: Our study not only provides a repertoire of lipid or cholesterol regulatory genes from multiple angles but also reveals a causal link between a gamma-glutamyltransferase and cholesterol metabolism.PMID:40098013 | DOI:10.1186/s13059-025-03531-8

EMBO Rep. 2025 Mar 17. doi: 10.1038/s44319-025-00416-6. Online ahead of print.ABSTRACTAberrant mitochondrial function has been associated with an increasingly large number of human disease states. Observations from in vivo models where mitochondrial function is altered suggest that maladaptations to mitochondrial dysfunction may underpin disease pathology. We hypothesized that the severity of this maladaptation could be shaped by the plasticity of the system when mitochondrial dysfunction manifests. To investigate this, we have used inducible fly models of mitochondrial complex I (CI) dysfunction to reduce mitochondrial function at two stages of the fly lifecycle, from early development and adult eclosion. Here, we show that in early life (developmental) mitochondrial dysfunction results in severe reductions in survival and stress resistance in adulthood, while flies where mitochondrial function is perturbed from adulthood, are long-lived and stress resistant despite having up to a 75% reduction in CI activity. After excluding developmental defects as a cause, we went on to molecularly characterize these two populations of mitochondrially compromised flies, short- and long-lived. We find that our short-lived flies have unique transcriptomic, proteomic and metabolomic responses, which overlap significantly in discrete models of CI dysfunction. Our data demonstrate that early mitochondrial dysfunction via CI depletion elicits a maladaptive response, which severely reduces survival, while CI depletion from adulthood is insufficient to reduce survival and stress resistance.PMID:40097814 | DOI:10.1038/s44319-025-00416-6

Nat Genet. 2025 Mar 17. doi: 10.1038/s41588-025-02135-z. Online ahead of print.ABSTRACTThe tea plant stands as a globally cherished nonalcoholic beverage crop, but the genetic underpinnings of important agronomic and metabolomic traits remain largely unexplored. Here we de novo deep resequenced 802 tea plants and their relative accessions globally. By integrating public Camellia accessions, we constructed a comprehensive genome-wide genetic variation map and annotated deleterious mutations for 1,325 accessions. Population genetic analyses provided insights into genetic divergence from its relatives, different evolutionary bottlenecks, interspecific introgression and conservation of wild relatives. Our findings suggest the pivotal role of southwest China as the origin of tea plants, revealing the genetic diversity and domestication status of ancient tea plants. Genome-wide association studies herein identified thousands of substantial associations with leaf shape and metabolite traits, pinpointing candidate genes for crucial agronomic and flavor traits. This study illuminates the tea plant's evolution and provides references for tea plant design breeding.PMID:40097782 | DOI:10.1038/s41588-025-02135-z

In Vitro Cell Dev Biol Anim. 2025 Mar 17. doi: 10.1007/s11626-025-01028-z. Online ahead of print.ABSTRACTCell Painting is a sophisticated high-content imaging technique that has been predominantly applied to mammalian cells. Recent advancements have extended its applicability to the first insect cell line, the ovarian cell line Sf9, revealing significant insights into similarities and differences in cellular responses between different taxonomic groups. This study explores the utility of Cell Painting in Helicoverpa zea gut-derived cells, specifically the RP-HzGUT-AW1 cell line, to assess the specifics of insect epithelial cells in response to chemical treatments. Upon adaptation of the analysis pipeline to accommodate their unique morphology and characteristics, our investigations revealed distinct responses of RP-HzGUT-AW1 cells compared to the ovarian insect cell line Sf9. Variations were obtained not only in the dose-response behavior to treatments but also in the overall detectability of specific modes of action. Specifically, processes that relate to osmoregulation and the formation of epithelial structures showed the most significant and distinct responses. This suggests that the specific morphological and physiological attributes of these gut-derived insect cells contribute to unique phenotypic profiles, which enables in-depth interpretation of drug efficacy and safety in these models.PMID:40097748 | DOI:10.1007/s11626-025-01028-z

Sci Rep. 2025 Mar 17;15(1):9063. doi: 10.1038/s41598-025-93710-1.ABSTRACTThis study aimed to investigate the effects of dietary supplementation with Selenium nanoparticles (SeNPs) on lactation performance, rumen microbial communities, and metabolism in dairy goats under heat stress conditions. Twenty Guanzhong dairy goats with the same parity, similar lactation period (120 ± 15 days), and similar milk yield (1.20 ± 0.16 kg/day) were randomly divided into two groups, with 10 replicates in each group. The control group was fed a standard diet, while the experimental group was supplemented with 0.5 mg SeNPs/kg DM based on the standard diet. The pretrial period lasted for seven days, followed by a 30-day trial period. The results showed that dietary supplementation with SeNPs significantly increased milk yield, milk fat and lactose content in dairy goats, under heat stress conditions. SeNPs significantly altered the composition of the rumen microbiota, increasing the relative abundance of Prevotella and Ruminococcus while decreasing the relative abundance of Succiniclasticum. This enhanced the rumen's ability to degrade starch and fiber under heat stress conditions. Non-targeted metabolomic analysis revealed a total of 119 differential metabolites between the two groups, indicating changes in rumen metabolism. Further correlation analysis indicated that Rumen bacterium R-21 was positively correlated with propionate, while Ralstonia insidiosa was negatively correlated with γ-glutamylcysteine. Additionally, several differential microbes, including Succinivibrio dextrinosolvens, Rummeliibacillus pycnus, Ralstonia insidiosa, and Prevotella sp BP1-56, were significantly correlated with milk composition. In conclusion, dietary supplementation with SeNPs can positively impact milk yield, milk components, and metabolism in dairy goats by improving the composition of the rumen microbiota under heat stress conditions.PMID:40097638 | DOI:10.1038/s41598-025-93710-1

Sci Rep. 2025 Mar 17;15(1):9084. doi: 10.1038/s41598-025-93426-2.ABSTRACTThe aging of seeds seriously affects their yield. Vacuum and low temperatures have been shown to prolong seed life and delay seed senescence. However, the underlying mechanisms that control these processes in safflower have yet to be explored. Therefore, this study aimed to study the structural, physiological and biochemical changes that occur in safflower seeds stored for one year at different temperatures and sealing conditions. X-ray imaging, germination percentage determination, determination of water content, and TTC staining were utilized to analyze seed structure and viability. The structure of its outer surface was observed by scanning electron microscope, and the changes of catalase activity and malondialdehyde content were determined to understand its physiological and biochemical status. In addition, lipidomic and proteomic analyses were performed. The results showed that the germination percentage was improved under vacuum and low temperature conditions. Compared with high-temperature storage, low-temperature storage not only reduces the level of reactive oxygen species, but also facilitates the preservation of intact seed structure. Lipidomic analysis indicated the levels of PA reduced at low temperatures, while the content of PC, PE, PS, and PG exhibited an inverse correlation, increasing as temperatures decreased. Proteomic analysis identified two proteins (HH-013791-RA, HH-017308-RA) that may be involved in fatty acid metabolism and carbon metabolism respectively. Expression levels of these proteins were found to be lower at -18 °C, but increased with increasing storage temperatures. Storing safflower seeds under low-temperature and vacuum conditions significantly enhances germination rates and preserves seed structure by reducing reactive oxygen species levels. Two proteins (HH-013791-RA, HH-017308-RA) in the fatty acid metabolism and carbon metabolism pathways are temperature-regulated, and are involved in lipid metabolism, affecting seed structure and vitality.PMID:40097562 | DOI:10.1038/s41598-025-93426-2

Sci Rep. 2025 Mar 18;15(1):9226. doi: 10.1038/s41598-025-93305-w.ABSTRACTRapid evaporative ionization mass spectrometry (REIMS) shows promise as a preparation-free tissue analysis tool with the prospect for real-time diagnostics. Given that hepatic steatosis is characterized by shifts in lipid species and abundance, we selected it as basis for method development, as REIMS specifically measures lipidomic profiles. However, further validation and protocol refinement are necessary to establish its clinical utility. In this study, we applied REIMS to steatotic human liver tissues, focusing on its ability to differentiate varying degrees of steatosis. We established standardized protocols for tissue handling and lipid analysis, which were essential for reliable data interpretation. Notably, our findings revealed that tissue size impacts REIMS sensitivity, with smaller samples yielding lower total ion counts and altered lipid profiles. Through principal component analysis, we identified key lipid classes, namely triacylglycerides, fatty acids, and glycerophospholipids. Despite a missing link between triacylglyceride abundance and degree of steatosis, we successfully identified condition-specific lipid patterns, with ceramides emerging as markers of advanced steatosis. Our study provides a protocol for the measurements of lipid standards showing the detailed degradation of specific lipids using iKnife-coupled REIMS. It highlights the pitfalls and limitations and provides critical recommendations for REIMS use. It also emphasizes the need for standardized biobanking and tissue preparation to ensure accurate lipid profiling, laying the groundwork for future protocol adjustments required for clinical application.PMID:40097529 | DOI:10.1038/s41598-025-93305-w

Clin Chem Lab Med. 2025 Mar 19. doi: 10.1515/cclm-2024-1360. Online ahead of print.ABSTRACTOBJECTIVES: Human tear analysis holds promise for biomarker discovery, but its clinical utility is hindered by the lack of standardized reference values, limiting interindividual comparisons. This study aimed at developing a protocol for normalizing metabolomic data from human tears, enhancing its potential for biomarker identification.METHODS: Tear metabolomic profiling was conducted on 103 donors (64 females, 39 males, aged 18-82 years) without ocular pathology, using the AbsoluteIDQ™ p180 Kit for targeted metabolomics. A predictive normalization model incorporating age, sex, and fasting time was developed to correct for interindividual variability. Key metabolites from six compound families (amino acids, biogenic amines, acylcarnitines, lysophosphatidylcholines, phosphatidylcholines, and sphingomyelins) were identified as normalization references. The approach was validated using Linear Discriminant Analysis (LDA) to test its ability to classify donor sex based on metabolite concentrations.RESULTS: Metabolite concentrations exhibited significant interindividual variability. The normalization model, which predicted metabolite concentrations based on a reference "concomitant" metabolite from each compound family, successfully reduced this variability. Using the ratio of observed-to-predicted concentrations, the model enabled robust comparisons across individuals. LDA classification of donor sex using acylcarnitine C4 achieved 78 % accuracy, correctly identifying 92 % of female donors. This approach outperformed traditional statistical and machine learning methods (Lasso logistic regression and Random Forest classification) in sex discrimination based on tear metabolomics.CONCLUSIONS: This novel normalization protocol significantly improves the reliability of tear metabolomics by enabling standardized interindividual comparisons. The approach facilitates biomarker discovery by mitigating variability in metabolite concentrations and may be extended to other biological fluids, enhancing its applicability in precision medicine.PMID:40097363 | DOI:10.1515/cclm-2024-1360

G3 (Bethesda). 2025 Mar 18:jkaf025. doi: 10.1093/g3journal/jkaf025. Online ahead of print.ABSTRACTUnsaturated fatty acids (UFAs) play crucial roles in various physiological and pathological processes. In animals, these lipids are synthesized from saturated fatty acids through the action of delta 9 (Δ9) desaturases. In C. elegans, three Δ9 desaturases are encoded by the genes fat-5, fat-6, and fat-7. The presence of multiple Δ9 desaturases has posed a significant challenge in developing a rapid and efficient approach to control UFA production in C. elegans and other model organisms. Utilizing the auxin-inducible degradation system, we specifically targeted the C. elegans fat-7 gene, responsible for the major stearoyl-CoA desaturase (SCD), while deleting fat-5 and fat-6. This design resulted in a strain that can be reversibly depleted of UFAs in the cells of interest. Conditional depletion in all somatic cells exhibited a pronounced auxin-dependent defect in UFA production. Using this system, we uncovered an essential requirement for de novo UFA production during L1 and L2 stages. Moreover, our results support a direct connection between UFA levels, fat storage and increased lipid turnover. This system will enable further studies exploring the cellular and physiological consequences of impairing UFA biosynthesis at different developmental stages or in specific tissues.PMID:40097139 | DOI:10.1093/g3journal/jkaf025

Metabolism. 2025 Mar 15:156191. doi: 10.1016/j.metabol.2025.156191. Online ahead of print.ABSTRACTLipedema is a chronic disease in females characterized by pathologic subcutaneous adipose tissue expansion and hitherto remains without druggable targets. In this observational study, we investigated the molecular hallmarks of lipedema using an unbiased multi-omics approach. We found adipokine dysregulation in lipedema patients participating in a cross-sectional clinical study (ClinicalTrial.gov, NCT02838277), pointing towards the adipocyte as a key player. Analyses of newly generated transcriptomic (SRA, PRJNA940039) and proteomic (ProteomeXchange, PXD058489) datasets of early- and late-stage lipedema samples revealed a local downregulation of factors involved in inflammation. Concomitantly, factors involved in cellular respiration, oxidative phosphorylation, as well as in mitochondrial organization were upregulated. Measuring a cytokine and chemokine panel in the serum of non-menopausal women, we observed little systemic changes in inflammatory markers, but a trend towards increased VEGF. Metabolomic and lipidomic analyses highlighted altered circulating glutamic acid, glutathione, and sphingolipid levels, suggesting a broader dysregulation of metabolic and inflammatory processes. We subsequently benchmarked a set of models to accurately predict lipedema using serum factor measurements (sLPM). Our study of the molecular signature of lipedema thus provides not only potential targets for therapeutic intervention, but also candidate markers of disease development and progression.PMID:40097137 | DOI:10.1016/j.metabol.2025.156191

Toxicology. 2025 Mar 15:154120. doi: 10.1016/j.tox.2025.154120. Online ahead of print.ABSTRACTPolychlorinated biphenyls (PCBs) remain an environmental health concern due to their persistence and ongoing release from legacy and emerging sources. 2,2',5,5'-Tetrachlorobiphenyl (PCB52), a PCB congener frequently detected in the environment and human blood, is oxidized to 2,2',5,5'-tetrachlorobiphenyl-4-ol (4-52). The neurotoxicity of this hydroxylated (OH-PCB) metabolite remains poorly characterized. In this study, we exposed 4-week-old male Sprague Dawley rats to 4-52 via a polymeric implant drug delivery system grafted in the subcutaneous cavity at 4-52 concentrations of 0%, 1%, 5%, and 10% in the implant (w/w) for 28 days. Metabolomic analyses were performed in the serum. RNA sequencing, immunofluorescence, and dopamine (DA) measurement with electrochemical detection were used to characterize the effects of 4-52 on the striatum and cerebellum, brain regions implicated in PCB neurotoxicity. Serum metabolomic analysis revealed disruptions in the "arginine biosynthesis" pathway following 4-52 exposure. Exposure to 4-52 caused moderate transcriptomic changes in pathways related to "oxidative phosphorylation" and "neuroactive ligand-receptor interactions." Immunofluorescence showed no significant alterations in microglial, astrocytic, or apoptotic biomarkers. In the medium dose group, the levels of the DA metabolite DOPAL (3,4-dihydroxyphenylacetaldehyde) were significantly reduced in the striatum. Subsequent multi-omics network analysis identified interactions among OH-PCBs, endogenous metabolites, and the transcriptome. For example, levels of glutamic acid, aspartic acid, choline, and glycerophosphocholine negatively correlated with 4-52 in the striatum. Expression levels of heat shock protein (HSP) family genes, Hsp90b1, Hspa8, and Hspa5, positively correlated with serum metabolites, including proline, 1-methylguanidine, and methionine sulfoxide. These findings identify novel biomarkers and targets of 4-52-induced neurotoxicity.PMID:40097135 | DOI:10.1016/j.tox.2025.154120

Mol Metab. 2025 Mar 15:102121. doi: 10.1016/j.molmet.2025.102121. Online ahead of print.ABSTRACTThe Glucagon-like peptide-2 (GLP-2) analogue teduglutide is used clinically for the treatment of short bowel syndrome and intestinal failure occurring after extensive intestinal resection. A recently discovered effect of GLP-2 treatment is the inhibition of gallbladder motility and increased gallbladder refilling. However, the impact of these two GLP-2-characteristic effects on bile acid metabolism in health and after intestinal resection is not well characterized. To study effects of teduglutide treatment, we combined the selenium-75-homocholic acid taurine (SeHCAT) assay with novel spatial imaging in healthy mice and after ileocecal resection (ICR mice) and associated the results with clinical stage targeted bile acid metabolomics as well as gene expression analyses. ICR mice had virtual complete intestinal loss of secondary bile acids, and an increased ratio of 12α-hydroxylated vs. non-12α-hydroxylated bile acids, which was attenuated by teduglutide. Teduglutide promoted SeHCAT retention in healthy and in ICR mice. Acute concentration of the SeHCAT-signal into the hepatobiliary system was observed. Teduglutide induced significant repression of hepatic cyp8b1 expression, which was associated with induction of MAF BZIP Transcription Factor G. The data suggest that GLP-2-pharmacotherapy in mice significantly slows bile acid circulation primarily via hepatic Farnesoid X receptor-signaling.PMID:40097102 | DOI:10.1016/j.molmet.2025.102121