Metabolites

17 pages, 1366 KiB

Open AccessArticle

¹H NMR-Based Analysis to Determine the Metabolomics Profile of Solanum nigrum L. (Black Nightshade) Grown in Greenhouse Versus Open-Field Conditions

by Lufuno Ethel Nemadodzi, Gudani Millicent Managa and Ndivho Nemukondeni

Metabolites 2025, 15(5), 344; https://doi.org/10.3390/metabo15050344 - 21 May 2025

Viewed by 478

Abstract

Background: Equally with other indigenous green leafy vegetables, Solunum nigrum L. has been widely consumed by the VhaVenda tribe found in the Limpopo Province of South Africa since ancient times as a source of food diversification due to its higher-quality nutritional value, sustainability, [...] Read more.

Background: Equally with other indigenous green leafy vegetables, Solunum nigrum L. has been widely consumed by the VhaVenda tribe found in the Limpopo Province of South Africa since ancient times as a source of food diversification due to its higher-quality nutritional value, sustainability, food security, and medicinal benefits. It is mostly cultivated from seeds in seedling trays and transplanted in the open field, and at the maturity stage, marketing and distribution are mainly conducting through informal markets (i.e., street vendors). However, recently, it can be found in selected supermarkets and commercial grocery stores in South Africa. The leaves and young shoots of S. nigrum are cooked solely and/or as a supplementary vegetable with Brassica rapa L. subsp. chinensis (Chinese cabbage), Spinacia oleracea L. (spinach), Amaranthus graecizans L. (green amaranth), Solanum lycopersicum L. (tomato), and/or cooking oil for flavor. Objective: Contrary to other green leafy vegetables, few studies have been conducted on the metabolites released by S. nigrum and the influence of growing conditions on the metabolites thereof. Method: A ¹H-nuclear magnetic resonance tool was used to identify the untargeted metabolites released by S. nigrum, and spectra were phase-corrected and binned with MestReNova and statistically analyzed with SIMCA 18.0.2. Results: The findings showed that a total of 12 metabolites were detected between the growing conditions. Eleven similar metabolites, such as glycocholate, chlorogenate (human health benefits), caffeine for its bitter taste, choline, 3-Chlorotyrosine (antidiabetic, blood pressure), etc., and a few vital soluble sugars, were detected in S. nigrum samples grown in the open field and greenhouse-cultivated. Glucose was exclusively detected in the S. nigrum grown under greenhouse conditions. Full article

(This article belongs to the Special Issue Unlocking Nature's Toolbox: Advances in Plant Metabolites Identification)

► Show Figures

Figure 1

18 pages, 1054 KiB

Open AccessArticle

Association Between Bioimpedance-Determined Metabolic Age and MASLD Risk Scores in Spanish Workers

by Ignacio Ramírez-Gallegos, Carla Busquets-Cortes, Hernán Paublini, Ángel Arturo López-González, Emilio Martínez-Almoyna-Rifá, Pedro Juan Tárraga López and José Ignacio Ramírez-Manent

Metabolites 2025, 15(5), 343; https://doi.org/10.3390/metabo15050343 - 21 May 2025

Viewed by 532

Abstract

Background: Metabolic dysfunction-associated steatotic liver disease (MASLD) is a prevalent liver disorder with significant metabolic implications. Metabolic age, determined through bioimpedance analysis, has emerged as a potential indicator of overall metabolic health. The objective of this study is to evaluate the association [...] Read more.

Background: Metabolic dysfunction-associated steatotic liver disease (MASLD) is a prevalent liver disorder with significant metabolic implications. Metabolic age, determined through bioimpedance analysis, has emerged as a potential indicator of overall metabolic health. The objective of this study is to evaluate the association between metabolic age and MASLD risk scores in a cohort of Spanish workers. Methods: A cross-sectional study was conducted on 8590 Spanish workers who underwent annual occupational health examinations between 2019 and 2020. Metabolic age was determined using bioelectrical impedance analysis, and the Avoidable Lost Life Years (ALLY) index was calculated as the difference between their metabolic and chronological age. MASLD risk was assessed using various validated scales, including the Fatty Liver Index (FLI), Hepatic Steatosis Index (HSI), Zhejiang University Index (ZJU), Fatty Liver Disease Index (FLD), and Lipid Accumulation Product (LAP). A multinomial logistic regression analysis was performed to examine the association between metabolic age and MASLD risk scores, adjusting for sociodemographic and lifestyle variables. Results: Higher metabolic age values were observed in individuals with greater MASLD risk across all evaluated scales. The mean metabolic age was consistently lower in women compared to men, and these differences were statistically significant (p < 0.01). Multinomial logistic regression analysis revealed that the strongest associations with increased metabolic age were found for MASLD risk scores, physical inactivity, and poor adherence to the Mediterranean diet. ROC curve analysis demonstrated a high predictive capacity for the FLD (AUC: 0.935 in women and 0.917 in men) and FLI (AUC: 0.900 in women and 0.833 in men), with high Youden index values. Conclusions: Metabolic age is significantly associated with MASLD risk, suggesting its potential as a non-invasive biomarker for identifying individuals with a higher risk for metabolic liver disease. Lifestyle factors, including physical activity and dietary patterns, play a crucial role in modulating metabolic age, highlighting the importance of targeted interventions for MASLD prevention. Further research is warranted to validate metabolic age as a prognostic tool in MASLD risk assessment. Full article

(This article belongs to the Section Endocrinology and Clinical Metabolic Research)

► Show Figures

Graphical abstract

13 pages, 1982 KiB

Open AccessArticle

Lipid Subclasses Differentiate Insulin Resistance by Triglyceride–Glucose Index

by Khaled Naja, Najeha Anwardeen, Omar Albagha and Mohamed A. Elrayess

Metabolites 2025, 15(5), 342; https://doi.org/10.3390/metabo15050342 - 20 May 2025

Viewed by 443

Abstract

Background: Insulin resistance is a key driver of metabolic syndrome and related disorders, yet its underlying metabolic alterations remain incompletely understood. The Triglyceride–Glucose (TyG) index is an emerging, accessible marker for insulin resistance, with growing evidence supporting its clinical utility. This study aimed [...] Read more.

Background: Insulin resistance is a key driver of metabolic syndrome and related disorders, yet its underlying metabolic alterations remain incompletely understood. The Triglyceride–Glucose (TyG) index is an emerging, accessible marker for insulin resistance, with growing evidence supporting its clinical utility. This study aimed to characterize the metabolic profiles associated with insulin resistance using the TyG index in a large, population-based cohort, and to identify metabolic pathways potentially implicated in insulin resistance. Methods: Here, we conducted a cross-sectional study using data from the Qatar Biobank, including 1255 participants without diabetes classified as insulin-sensitive or insulin-resistant based on TyG index tertiles. Untargeted serum metabolomics profiling was performed using high-resolution mass spectrometry. Our statistical analyses included orthogonal partial least squares discriminate analysis and linear models. Results: Distinct metabolic signatures differentiated insulin-resistant from insulin-sensitive participants. Phosphatidylethanolamines, phosphatidylinositols, and phosphatidylcholines, were strongly associated with insulin resistance, while plasmalogens and sphingomyelins were consistently linked to insulin sensitivity. Conclusions: Lipid-centric pathways emerge as potential biomarkers and therapeutic targets for the early detection and personalized management of insulin resistance and related metabolic disorders. Longitudinal studies are warranted to validate causal relationships. Full article

(This article belongs to the Special Issue Risk Prediction and Novel Biomarker Discovery of Chronic Diseases by Advanced Methodologies in Lipidomics and Metabolomics)

► Show Figures

Figure 1

18 pages, 2633 KiB

Open AccessArticle

Effects of Exercise on Gut Microbiome and Serum Metabolomics in Post-Traumatic Osteoarthritis Rats

by Xiaoxia Hao, Xingru Shang, Yiwen Zhang, Wenjie Hou, Ruimin Chi, Chunran Pan, Jiawei Liu, Xiaofeng Deng, Jiaming Zhang and Tao Xu

Metabolites 2025, 15(5), 341; https://doi.org/10.3390/metabo15050341 - 20 May 2025

Viewed by 432

Abstract

Objective: The aim of this work is to investigate the impact of exercise on gut microbiome composition, serum metabolites, and their correlation with osteoarthritis (OA) severity. Methods: Thirty-six Sprague-Dawley (SD) rats were randomly divided into four groups: Sham rats without treadmill walking (Sham/Sed [...] Read more.

Objective: The aim of this work is to investigate the impact of exercise on gut microbiome composition, serum metabolites, and their correlation with osteoarthritis (OA) severity. Methods: Thirty-six Sprague-Dawley (SD) rats were randomly divided into four groups: Sham rats without treadmill walking (Sham/Sed group, n = 9), Sham rats with treadmill walking 2 months (Sham/TW2M group, n = 9), PTOA rats without treadmill walking (PTOA/Sed group, n = 9), and PTOA rats with treadmill walking 2 months (PTOA/TW2M group, n = 9). The PTOA model was induced by transection of the anterior cruciate ligament (ACLT) and destabilization of the medial meniscus (DMM). Histological evaluation and micro-CT analysis were performed to observe the pathological changes in cartilage and subchondral bone, respectively. Additionally, we conducted 16S rDNA sequencing of fecal samples and untargeted metabolomic analysis using liquid chromatography–mass spectrometry (LC–MS) of serum samples to detect the alteration of gut microbiota composition and metabolites. Results: Exercise effectively mitigated OA-related pathological changes, including articular cartilage degeneration and subchondral bone loss. Moreover, 16S rDNA sequencing analysis of gut microbiome revealed a decreased abundance of Bacteroidetes (p < 0.01), Bacteroidia (p < 0.01), Rikenellaceae (p < 0.01), [Paraprevotellaceae] (p < 0.01), and Paraprevotella (p < 0.01) but an increase in Firmicutes (p < 0.01) in PTOA/TW2M group rats compared with PTOA/Sed group as a response to exercise. In addition, the results of metabolomics analysis showed that exercise treatment contributed to the upregulation of Daidzein and Anthranilic acid and downregulation of 1-Palmitoyllysophosphatidylcholine. Moreover, the correlation analysis showed that Rikenellaceae significantly positively correlated with both OARSI (r = 0.81, p < 0.01) and Mankin score (r = 0.83, p < 0.01) and negatively correlated with the serum level of Anthranilic acid (r = −0.56, p < 0.01) and Daidzein (r = −0.46, p < 0.01). Conclusions: Exercise can effectively mitigate OA through slowing down articular cartilage degeneration and subchondral bone loss, modulating gut microbiota composition, and increasing beneficial metabolites. Full article

(This article belongs to the Special Issue Dysbiosis and Metabolic Disorders of the Microbiota)

► Show Figures

Figure 1

15 pages, 1429 KiB

Open AccessArticle

Characterizing Plasma-Based Metabolomic Signatures for Metastasis in Non-Small Cell Lung Cancer

by Manlu Liu, Yanlong Zhu, Sean J. McIlwain, Haotian Deng, Allan R. Brasier, Ying Ge, Michelle E. Kimple and Andrew M. Baschnagel

Metabolites 2025, 15(5), 340; https://doi.org/10.3390/metabo15050340 - 20 May 2025

Viewed by 527

Abstract

Background/Objectives: The current staging of non-small cell lung cancer (NSCLC) relies on conventional imaging, which lacks the sensitivity to detect micrometastatic disease. The functional assessment of NSCLC progression may provide independent information to enhance the prediction of metastatic risk. The objective of [...] Read more.

Background/Objectives: The current staging of non-small cell lung cancer (NSCLC) relies on conventional imaging, which lacks the sensitivity to detect micrometastatic disease. The functional assessment of NSCLC progression may provide independent information to enhance the prediction of metastatic risk. The objective of this study was to determine if we could identify a metabolomic signature predictive of metastasis in patients with NSCLC treated with definitive radiation. Methods: Plasma samples were collected prospectively from patients enrolled in a clinical trial with non-metastatic NSCLC treated with definitive radiation. Metabolites were extracted, and mass spectrometry-based analysis was performed using a flow injection electrospray (FIE)–Fourier transform ion cyclotron resonance (FTICR) mass spectrometry (MS) method. Early metastasis was defined as metastasis within 1 year of radiation treatment. Results: The study cohort included 28 patients. FIE-FITCR produced highly reproducible profiles in technical replicates. A total of 51 metabolic features were identified to be different in patients with early metastasis compared to patients without early metastasis (all adjusted p-values < 0.05, Welch’s t-test), including glycerophospholipids, sphingolipids, and fatty acyls. In the follow-up samples collected after the initiation of chemotherapy and radiation treatment, a total of 174 metabolic features were significantly altered in patients who developed early metastasis compared to those who did not. Conclusions: We identified several distinct changes in the metabolic profiles of patients with NSCLC who developed metastatic disease within 1 year of definitive radiation. These findings highlight the potential of metabolomic profiling as a predictive tool for assessing metastatic risk in NSCLC. Full article

(This article belongs to the Special Issue Cancer Metabolomics 2024)

► Show Figures

Graphical abstract

18 pages, 12162 KiB

Open AccessArticle

Cadmium Exposure Disrupts Uterine Energy Metabolism and Coagulation Homeostasis During Labor in Institute of Cancer Research Mice: Insights from Transcriptomic Analysis

by Yueyang Wang, Yichen Bai, Yi Wang and Yan Cai

Metabolites 2025, 15(5), 339; https://doi.org/10.3390/metabo15050339 - 20 May 2025

Viewed by 556

Abstract

Background: Cadmium (Cd) is a highly toxic heavy metal. There are very few studies about the effects of Cd on reproductive health and metabolism, and even fewer on metabolic disorders in the uterus of mice in labor. This study is the first to [...] Read more.

Background: Cadmium (Cd) is a highly toxic heavy metal. There are very few studies about the effects of Cd on reproductive health and metabolism, and even fewer on metabolic disorders in the uterus of mice in labor. This study is the first to establish a model of Cd exposure in the uterus of laboring mice and investigate the underlying metabolic mechanisms through transcriptomic analysis. Methods: Pregnant mice received intraperitoneal injections of CdCl₂ (1.5 mg/kg) on gestational days 12.5, 14.5, and 16.5 were set up as the experimental group (Cd group), and pregnant mice injected with saline were set up as the control group (CT group). A total of 738 differentially expressed genes (DEGs) were screened using DESeq2 software, including 326 upregulated genes and 412 downregulated genes. Results: Through enrichment databases including the KEGG, GO, Reactome, and PANTHER, we identified 76 metabolism-related DEGs and performed protein–protein interaction (PPI) network analysis. The PPI results were visualized using Cytoscape software and further analyzed, with 18 hub genes (maximum clique centrality score > 10) identified through the MCC algorithm of the Cytohubba plugin. The results showed that the highest-scoring hub genes included mt-Co2, mt-Co3, mt-Atp6, mt-Atp8, mt-Nd3, and mt-Nd4l, which are involved in mitochondrial energy metabolism. The remaining lower-scoring hub genes were primarily associated with coagulation processes. Pathway analysis revealed hub genes predominantly involved in oxidative phosphorylation, complement and coagulation cascades, the cGMP-PKG signaling pathway, and thermogenesis. Conclusion: This study successfully established a Cd exposure-induced uterine injury model, providing valuable references for human reproductive health research. Full article

(This article belongs to the Special Issue Metabolic Responses to Environmental Challenges: Insights into Cellular Mechanism and Protective Strategy)

► Show Figures

Graphical abstract

16 pages, 2430 KiB

Open AccessArticle

A Comparative Analysis of Raw and Bran-Fried Acori tatarinowii Rhizoma Based on the Intelligent Sensory Evaluation System

by Yingna Le, Zhongjian Yang, Ruiping Wang, Shaolong Ma, Yang Cui, Kun Shi, Li Xin, Jinlian Zhang and Lingyun Zhong

Metabolites 2025, 15(5), 338; https://doi.org/10.3390/metabo15050338 - 20 May 2025

Viewed by 442

Abstract

Objectives: The study aimed to investigate the differences in odor, color, and taste characteristics between raw and bran-fried Acori tatarinowii Rhizoma (RATR and BATR) using advanced sensory evaluation technologies. The objective was to establish a reliable differential analysis method for distinguishing RATR and [...] Read more.

Objectives: The study aimed to investigate the differences in odor, color, and taste characteristics between raw and bran-fried Acori tatarinowii Rhizoma (RATR and BATR) using advanced sensory evaluation technologies. The objective was to establish a reliable differential analysis method for distinguishing RATR and BATR slices to support quality control in herbal processing. Methods: The Heracles NEO ultra-fast gas-phase electronic nose was employed to analyze odor profiles, while electronic eye and electronic tongue technologies were used to assess color and taste differences, respectively. Odor fingerprint analysis identified key volatile components, and colorimetric and taste measurements were conducted to compare RATR and BATR samples. Results: Fifteen characteristic odor components were identified, with methanol, 2-propanol, and 2-cyclopentenone potentially serving as discriminant markers differentiating RATR and BATR. PCA demonstrated exceptional separation efficacy, with a cumulative contribution rate of 99.937% for the primary components. Conclusions: The integration of Heracles NEO electronic nose, electronic eye, and electronic tongue technologies effectively distinguished RATR from BATR. This approach provides a novel strategy for online quality monitoring in herbal slice production and offers a robust analytical framework for the identification and quality assessment of processed herbal medicines. Full article

(This article belongs to the Section Food Metabolomics)

► Show Figures

Figure 1

17 pages, 2378 KiB

Open AccessArticle

Eliciting Clavulanic Acid Biosynthesis: The Impact of Bacillus velezensis FZB42 on the Metabolism of Streptoyces clavuligerus ATCC 27064

by Luisa F. Patiño, Carlos Caicedo-Montoya, Laura Pinilla-Mendoza, Jaison H. Cuartas and Rigoberto Ríos-Estepa

Metabolites 2025, 15(5), 337; https://doi.org/10.3390/metabo15050337 - 19 May 2025

Viewed by 468

Abstract

Background/Objectives: Clavulanic acid (CA) is produced by cell suspension cultures of Streptomyces clavuligerus ATCC 27064, and is widely used as a beta-lactamase inhibitor to combat antibiotic resistance. CA titers are moderate due to bioprocess complexity, prompting ongoing efforts to overcome these limitations. Methods: [...] Read more.

Background/Objectives: Clavulanic acid (CA) is produced by cell suspension cultures of Streptomyces clavuligerus ATCC 27064, and is widely used as a beta-lactamase inhibitor to combat antibiotic resistance. CA titers are moderate due to bioprocess complexity, prompting ongoing efforts to overcome these limitations. Methods: In this study, we aimed to evaluate the effect of live and inactivated Bacillus velezensis FZB42 cells on CA production in S. clavuligerus, and to explore the transcriptional response underlying this interaction using RNA-seq technology. Results: The addition of dead and live cells of B. velezensis improved CA production by 1.4 and 2.0-fold, respectively. Furthermore, the transcriptome of S. clavuligerus, obtained with live cells of B. velezensis FZB42 at the peak of maximum CA production, revealed that 410 genes were up-regulated and 594 were down-regulated under these conditions, with a padj < 0.05. Most of the genes from the cephamycin C and CA clusters were up-regulated, which correlates well with the increase in CA production. Likewise, S. clavuligerus ATCC 27064 enhanced the expression of genes encoding enzymes that scavenge endogenous H₂O₂, as well as other genes related to oxidative stress defense. Regarding downregulated genes, we found that S. clavuligerus decreased the expression of genes involved in the biosynthesis of terpenoids, polyketides, and lantibiotics, as well as the expression of the operon involved in the synthesis of the pyrroloquinoline quinone (PQQ) cofactor. Conclusions: These findings contribute to the understanding of S. clavuligerus metabolism and pave the way for future metabolic engineering efforts aimed at obtaining CA-overproducing strains. Full article

(This article belongs to the Section Microbiology and Ecological Metabolomics)

► Show Figures

Figure 1

16 pages, 3225 KiB

Open AccessArticle

Mechanism Study on the Preventive Effect of ELITEA Compound Tea on Hyperuricemia in Rats Based on Serum Untargeted Metabolomics

by Shulian Liu, Yongliang Zhu, Wenjiong Wang, Xianghan Zhang, Linrui Gao and Xiangjun Qiu

Metabolites 2025, 15(5), 336; https://doi.org/10.3390/metabo15050336 - 19 May 2025

Viewed by 348

Abstract

Background/Objectives: Hyperuricemia (HUA) has become the second largest metabolic disease after diabetes, and has become a major public health problem. The ELITEA compound tea extract can effectively reduce the serum uric acid level in HUA rat models. In this study, the mechanism [...] Read more.

Background/Objectives: Hyperuricemia (HUA) has become the second largest metabolic disease after diabetes, and has become a major public health problem. The ELITEA compound tea extract can effectively reduce the serum uric acid level in HUA rat models. In this study, the mechanism of ELITEA compound tea on HUA was analyzed through serum untargeted metabolomics analysis. Methods: The rat model of HUA was established by feeding rats with a high uric acid diet. A total of 24 male SD rats were divided into a blank control group, a hyperuricemia model group, and an ELITEA compound tea prevention experimental group. UPLC-MS/MS was used to detect changes in metabolites in the blood of the three groups of rats, in order to identify potential biomarkers and study the mechanism of ELITEA compound tea in preventing HUA. Results: The ELITEA compound tea exhibited significant preventive effects on HUA rats. The analysis results showed that after ELITEA combined tea intervention, the 257 metabolites downregulated in the HUA model group showed an upward trend. Meanwhile, the 115 metabolites upregulated in the HUA model group showed a decreasing trend. Six main enrichment pathways were obtained, including arginine biosynthesis, histidine metabolism, pyrimidine metabolism, tryptophan metabolism, vitamin B6 metabolism, arginine and proline metabolism. Conclusions: ELITEA compound tea can effectively reduce the serum uric acid levels in HUA model rats. Based on the in-depth analysis of untargeted metabolomics, ELITEA compound tea mainly regulates the arginine biosynthesis pathway by modulating three important metabolites, arginine, glutamate, and ornithine, to reduce serum uric acid. Full article

(This article belongs to the Section Animal Metabolism)

► Show Figures

Figure 1

42 pages, 830 KiB

Open AccessReview

Gut–Brain Inflammatory Pathways in Attention-Deficit/Hyperactivity Disorder: The Role and Therapeutic Potential of Diet

by Naomi Lewis, Jim Lagopoulos and Anthony Villani

Metabolites 2025, 15(5), 335; https://doi.org/10.3390/metabo15050335 - 19 May 2025

Viewed by 1518

Abstract

Attention-deficit/hyperactivity disorder (ADHD) is a common childhood-onset neurodevelopmental disorder that often persists into adulthood, leading to various adverse outcomes. Its underlying pathology is multifactorial, involving neurotransmitter imbalances, gut microbiota alterations, and oxidative and inflammatory dysregulation. Diet, a key environmental modifier of gut ecology, [...] Read more.

Attention-deficit/hyperactivity disorder (ADHD) is a common childhood-onset neurodevelopmental disorder that often persists into adulthood, leading to various adverse outcomes. Its underlying pathology is multifactorial, involving neurotransmitter imbalances, gut microbiota alterations, and oxidative and inflammatory dysregulation. Diet, a key environmental modifier of gut ecology, is consistently poorer in individuals with ADHD, with multiple nutrients implicated in its pathophysiology. This review examines the role of specific nutrients such as omega-3 fatty acids, key micronutrients, and potentially harmful dietary components, as well as broader dietary patterns, particularly the Western diet and Mediterranean diet (MedDiet), in relation to ADHD symptoms. It also evaluates both whole-diet and supplement-based clinical interventions, supporting the growing recognition of nutrition as a safe and relatively affordable modifiable factor in ADHD management. Additionally, the biological mechanisms linking diet to ADHD are reviewed, highlighting strong evidence for the involvement of gut dysbiosis and inflammatory processes. Despite the well-documented antioxidant, anti-inflammatory, and microbiome benefits of the MedDiet, direct research investigating its role in ADHD remains limited. Most whole-diet approaches to date have focused on elimination diets, leaving a significant gap in understanding the potential role of the MedDiet in ADHD management. Therefore, this review outlines preliminary evidence supporting the MedDiet and its key components as modulators of ADHD-related biological pathways, indicating its potential as a therapeutic approach. However, further research is required to rigorously evaluate its clinical efficacy. Finally, the limitations of observational and interventional nutritional research in ADHD are discussed, along with recommendations for future research directions. Full article

(This article belongs to the Special Issue The Roles of Diet, Gut Microbiome and Microbial Metabolome in Precision Nutrition)

► Show Figures

Figure 1

19 pages, 9097 KiB

Open AccessArticle

Metabolic Markers Demonstrate the Heterogeneity of Walking Ability in Non-Disabled Community-Dwelling Older Adults

by Shanshan Yao, Ziling Mao, Megan M. Marron, Eleanor M. Simonsick, Venkatesh L. Murthy, Ravi V. Shah and Anne B. Newman

Metabolites 2025, 15(5), 334; https://doi.org/10.3390/metabo15050334 - 19 May 2025

Viewed by 436

Abstract

Background: Walking ability is important for the quality of life of older adults. A self-reported walking ability index (WAI) covering the difficulty and ease of walking captures a broader spectrum of walking ability in healthy older persons. Methods: Using metabolomics in [...] Read more.

Background: Walking ability is important for the quality of life of older adults. A self-reported walking ability index (WAI) covering the difficulty and ease of walking captures a broader spectrum of walking ability in healthy older persons. Methods: Using metabolomics in the Health, Aging and Body Composition study, we identified Year 2 metabolites cross-sectionally and longitudinally related to WAI (0–9, higher scores indicate better walking ability) using probabilistic index models and multinomial logistic models, respectively. Results: Among 2334 participants (mean age 74.6 years, 51% women, 37% Black), 27% scored 0–5, 36% scored 6–8, and 37% scored 9 at Year 2. Over 4 years, 52% maintained a stable WAI, 6% improved, while 42% declined (22% 1–2 points and 20% >2 points decline). We identified 81 metabolites significantly associated with both poorer concurrent WAI and faster decline, including higher acylcarnitine species, shorter-chain saturated diglycerides and triglycerides, and TCA cycle intermediates (cis-aconitic, fumaric, and malic acids), and lower phospholipids levels. Eighteen additional metabolites were only associated with faster WAI decline: higher short-chain saturated triglycerides and energy metabolism markers (ATP/ADP/AMP) and lower margaric acid and glycine levels. Notably, those with improved WAI, despite poorer baseline WAI and lifestyles, showed more favorable metabolic profiles than others. Conclusions: Metabolites linked to the TCA cycle and energy metabolism, as well as inflammation and protein catabolism, were related to mobility function. Some metabolites might be particularly important for the early detection of older adults at risk of mobility decline. Metabolic profiles may also help identify older individuals (i.e., with improving WAI) with greater metabolic resilience to lifestyle risk factors and health conditions. Full article

(This article belongs to the Special Issue New Analytical Techniques and Applications of Metabolomics and Lipidomics)

► Show Figures

Figure 1

17 pages, 1684 KiB

Open AccessArticle

Dietary Nicotinamide Mononucleotide, a Key NAD⁺ Intermediate, Alleviates Body Fat Mass and Hypertriglyceridemia by Enhancing Energy Expenditure with Promotion of Fat Oxidation and Hepatic Lipolysis and Suppressing Hepatic Lipogenesis in db/db Mice

by Bungo Shirouchi, Sarasa Mitsuta, Mina Higuchi, Mai Okumura and Kazunari Tanaka

Metabolites 2025, 15(5), 333; https://doi.org/10.3390/metabo15050333 - 18 May 2025

Viewed by 682

Abstract

Background/Objectives: Supplementation with nicotinamide mononucleotide (NMN), a key nicotinamide adenine dinucleotide (NAD⁺) intermediate, exerts anti-aging, anti-obesity, and anti-diabetic effects in animal experiments. However, previous studies have evaluated NMN supplementation using oral administration in drinking water or by intraperitoneal administration. No [...] Read more.

Background/Objectives: Supplementation with nicotinamide mononucleotide (NMN), a key nicotinamide adenine dinucleotide (NAD⁺) intermediate, exerts anti-aging, anti-obesity, and anti-diabetic effects in animal experiments. However, previous studies have evaluated NMN supplementation using oral administration in drinking water or by intraperitoneal administration. No studies have reported whether NMN exerts beneficial effects when incorporated into the diet. The diet is a multicomponent mixture of many nutrients that may interact with each other, thus weakening the effects of NMN. In the present study, we evaluated whether dietary NMN intake protects obese diabetic db/db mice from obesity-related metabolic disorders, such as dyslipidemia, hepatic steatosis, hyperglycemia, and hyperinsulinemia. Methods: Five-week-old male db/db mice were randomly assigned to two groups and fed for four weeks either a control diet containing 7% corn oil and 0.1% cholesterol (CON group, n = 6) or a diet supplemented with 0.5% NMN (NMN group, n = 5). Results: After 4 weeks of feeding, dietary NMN intake alleviated obesity, hypertriglyceridemia, and hepatic triglyceride accumulation in db/db mice. Respiratory gas analysis indicated that dietary NMN intake significantly enhanced energy expenditure by suppressing carbohydrate oxidation and increasing fat oxidation after 3 weeks of feeding. Additionally, the suppression of the increase in plasma triglyceride (TG) levels by dietary NMN intake was attributable to a reduction in hepatic TG levels through the suppression of fatty acid synthesis and the enhancement of fatty acid β-oxidation in the liver. Furthermore, the improvement in hepatic fatty acid metabolism induced by dietary NMN intake was partially responsible for the significant increase in plasma adiponectin and soluble T-cadherin levels. Conclusions: This is the first report to show that dietary NMN intake but not oral administration in drinking water or intraperitoneal administration alleviates body fat mass and hypertriglyceridemia by enhancing energy expenditure, with preferential promotion of fat oxidation, the enhancement of hepatic lipolysis, and the suppression of hepatic lipogenesis in db/db mice. Full article

(This article belongs to the Special Issue Lipid Metabolism Dysregulation in Metabolic Disorders: Unraveling the Molecular Complexity)

► Show Figures

Graphical abstract

15 pages, 1477 KiB

Open AccessArticle

Urine Metabolomic Profiling and Machine Learning in Autism Spectrum Disorder Diagnosis: Toward Precision Treatment

by Shula Shazman, Julie Carmel, Maxim Itkin, Sergey Malitsky, Monia Shalan, Eyal Soreq, Evan Elliott, Maya Lebow and Yael Kuperman

Metabolites 2025, 15(5), 332; https://doi.org/10.3390/metabo15050332 - 16 May 2025

Viewed by 957

Abstract

Background: Autism spectrum disorder (ASD) diagnosis traditionally relies on behavioral assessments, which can be subjective and often lead to delayed identification. Recent advances in metabolomics and machine learning offer promising alternatives for more objective and precise diagnostic approaches. Methods: First-morning urine samples were [...] Read more.

Background: Autism spectrum disorder (ASD) diagnosis traditionally relies on behavioral assessments, which can be subjective and often lead to delayed identification. Recent advances in metabolomics and machine learning offer promising alternatives for more objective and precise diagnostic approaches. Methods: First-morning urine samples were collected from 52 children (32 with ASD and 20 neurotypical controls), aged 5.04 ± 1.87 and 5.50 ± 1.74 years, respectively. Using liquid chromatography-mass spectrometry (LC-MS), 293 metabolites were identified and categorized into 189 endogenous and 104 exogenous metabolites. Various machine learning classifiers (random forest, logistic regression, random tree, and naïve Bayes) were applied to differentiate ASD and control groups through 10-fold cross-validation. Results: The random forest classifier achieved 85% accuracy and an area under the curve (AUC) of 0.9 using all 293 metabolites. Classification based solely on endogenous metabolites yielded 85% accuracy and an AUC of 0.86, whereas using exogenous metabolites alone resulted in lower performance (71% accuracy and an AUC of 0.72). Conclusion: This study demonstrates the potential of urine metabolomic profiling, particularly endogenous metabolites, as a complementary diagnostic tool for ASD. The high classification accuracy highlights the feasibility of developing assistive diagnostic methods based on metabolite profiles, although further research is needed to link these profiles to specific behavioral characteristics and ASD subtypes. Full article

(This article belongs to the Section Bioinformatics and Data Analysis)

► Show Figures

Figure 1

12 pages, 2945 KiB

Open AccessArticle

Synergistic Impact of Aerobic Exercise and Resveratrol on White Adipose Tissue Browning in Obese Rats: Mechanistic Exploration and Biological Insights

by Yulong Hu, Yihan Wu, Chunlong Wang, Qiguan Jin and Xianghe Chen

Metabolites 2025, 15(5), 331; https://doi.org/10.3390/metabo15050331 - 16 May 2025

Viewed by 470

Abstract

Obesity, marked by excessive white adipose tissue (WAT) accumulation, worsens metabolic disorders, and inducing WAT browning is a promising therapy. This study examined the synergistic effects of moderate-intensity aerobic training and resveratrol (RES) on WAT browning and its underlying mechanisms in obese male [...] Read more.

Obesity, marked by excessive white adipose tissue (WAT) accumulation, worsens metabolic disorders, and inducing WAT browning is a promising therapy. This study examined the synergistic effects of moderate-intensity aerobic training and resveratrol (RES) on WAT browning and its underlying mechanisms in obese male rats. Methods: Male Sprague Dawley rats were divided into a normal diet control group (n = 8) and a high-fat-diet modeling group (n = 32), with the rats in the latter group being further divided randomly in groups of eight into a high-fat group; a high-fat, exercise group; a high-fat, RES group; and a high-fat, exercise-combined-with-RES group. The rats in the exercise intervention groups underwent moderate-intensity aerobic treadmill exercise for one hour daily, six days a week, while those in the RES groups received a 50 mg/kg/d RES solution via gavage before exercise, once daily, six days a week. Both interventions lasted eight weeks. Results: The combined intervention synergistically suppressed weight gain and visceral fat accumulation. WAT browning was enhanced, evidenced by upregulated UCP1 and CIDEA expression. Mitochondrial biogenesis was activated via the SIRT1-PGC-1α-NRF-1-TFAM pathway, accompanied by elevated mitochondrial enzyme activity and improved lipid mobilization (reduced serum free fatty acids and triglycerides). Conclusions: The combination of aerobic exercise and RES promotes WAT browning and lipolysis by enhancing mitochondrial biogenesis and stimulating mitochondrial thermogenesis through the modulation of the SIRT1-PGC-1α-NRF-1-TFAM pathway. Full article

(This article belongs to the Section Lipid Metabolism)

► Show Figures

Figure 1

23 pages, 3896 KiB

Open AccessArticle

Storage Profiling: Evaluating the Effect of Modified Atmosphere Packaging on Metabolomic Changes of Strawberries (Fragaria × ananassa)

by Johannes Brockelt, Robin Dammann, Jennifer Griese, Agnes Weiss, Markus Fischer and Marina Creydt

Metabolites 2025, 15(5), 330; https://doi.org/10.3390/metabo15050330 - 15 May 2025

Viewed by 535

Abstract

Background/Objectives: Strawberries (Fragaria × ananassa) are among the most commonly consumed fruits due to their taste and nutritional benefits. However, their high rate of spoilage poses a major problem during the period from harvest and transport to further processing or marketing. [...] Read more.

Background/Objectives: Strawberries (Fragaria × ananassa) are among the most commonly consumed fruits due to their taste and nutritional benefits. However, their high rate of spoilage poses a major problem during the period from harvest and transport to further processing or marketing. The aim of this study was, therefore, to investigate the effects of passive modified atmosphere packaging on the metabolome and shelf life of strawberries as a more sustainable alternative compared to standard market storage conditions. Methods: A total of 99 strawberry samples were analyzed for microbial viable counts, water content, and metabolomic changes using non-targeted low-resolution near-infrared spectroscopy, high-resolution mass spectrometry, and microbial culture-based methods. Results: Using near-infrared spectroscopy as a rapid screening method, the linear regression model indicated that strawberries stored under modified atmosphere packaging conditions had a longer shelf life. Furthermore, lipidomic analysis using mass spectrometry showed that the levels of spoilage biomarkers, such as oxidized phosphatidylcholines and lysophosphatidylcholines, were increased under common market storage conditions without a controlled atmosphere. In contrast, the levels of these metabolites were reduced when strawberries were stored in modified atmosphere packaging. Moreover, the strawberries stored under modified atmosphere packaging had a lower number of bacteria, yeasts, and molds as well as a lower water loss throughout the entire storage period. Conclusions: Overall, the study highlights the potential of passive modified atmosphere packaging films to extend the shelf life and thus maintain the edibility of strawberries over a longer period. Full article

(This article belongs to the Special Issue Metabolomic Technology in Quality and Safety of Agricultural Products and Foods)

► Show Figures

Figure 1

16 pages, 5619 KiB

Open AccessArticle

Atractylenolide I Inhibits Nicotine-Induced Macrophage Pyroptosis and Alleviates Atherogenesis by Suppressing the TLR4/ROS/TXNIP/NLRP3 Pathway

by Huan-Huan Li, Xian Liu, Yu-Ping Wang, Xi Xu, Lin Zhu, Wei Zhang and Kun Ren

Metabolites 2025, 15(5), 329; https://doi.org/10.3390/metabo15050329 - 15 May 2025

Cited by 1 | Viewed by 515

Abstract

Background/Objectives: Studies have shown that Atractylenolide I (AT-I) can exert anti-inflammatory and anti-oxidative effects, protecting against the development of various kinds of cardiovascular diseases. However, whether AT-I prevents nicotine-induced atherogenesis is unknown. This study was designed to explore the effects of AT-I on [...] Read more.

Background/Objectives: Studies have shown that Atractylenolide I (AT-I) can exert anti-inflammatory and anti-oxidative effects, protecting against the development of various kinds of cardiovascular diseases. However, whether AT-I prevents nicotine-induced atherogenesis is unknown. This study was designed to explore the effects of AT-I on nicotine-induced macrophage pyroptosis and the progression of atherosclerosis. Methods: RT-qPCR and Western blot were used to detect the mRNA and protein levels of TXNIP and pyroptosis-related factors in THP-1-derived macrophages. ELISA was used to detect the secretion of pro-inflammatory cytokines. Hoechst/PI double-staining assay was used to assess plasma membrane integrity. The ROS assay kit, LDH release assay kit, and caspase-1 activity assay kit were used to detect ROS production, LDH release, and caspase-1 activity. Oil Red O, HE, and Masson staining were used to evaluate lipid accumulation, lesion size, and plaque stability in HFD-fed apoE^−/− mice. Results: AT-I treatment significantly decreased pyroptosis-related factors expression, disrupted plasma membrane integrity, and down-regulated pro-inflammatory cytokines secretion, thereby inhibiting nicotine-induced pyroptosis of THP-1-derived macrophages. In addition, AT-I decreased ROS production and the expression of TLR4 and TXNIP. Lentivirus overexpression of TLR4 or TXNIP, or pre-treatment with ROS agonist, mainly reversed the anti-pyroptotic effects of AT-I in nicotine-treated THP-1-derived macrophages. Additionally, administering AT-I to HFD-fed apoE^−/− mice markedly decreased nicotine-induced up-regulation of pyroptosis-related proteins in the aortas. Enzymatic methods and ELISA assay suggested that AT-I improved dyslipidemia and inflammation in vivo. Oil Red O, HE, and Masson staining showed that AT-I alleviated lipid accumulation, decreased plaque size, and increased plaque stability. Conclusions: Taken together, AT-I can be regarded as a potential phytomedicine that protects against macrophage pyroptosis and atherosclerosis triggered by nicotine. Full article

(This article belongs to the Special Issue Metabolic Responses to Environmental Challenges: Insights into Cellular Mechanism and Protective Strategy)

► Show Figures

Figure 1

27 pages, 1935 KiB

Open AccessReview

Staphylococcus aureus Endocarditis Immunothrombosis

by Francesco Nappi

Metabolites 2025, 15(5), 328; https://doi.org/10.3390/metabo15050328 - 15 May 2025

Viewed by 593

Abstract

Background: Infective endocarditis continues to represent a challenge for healthcare systems, requiring careful management and resources. Recent studies have indicated a shift in the predominant pathogens of concern, with Streptococcus sp. a being superseded by Staphylococcus sp. and Enterococcus sp. as the leading [...] Read more.

Background: Infective endocarditis continues to represent a challenge for healthcare systems, requiring careful management and resources. Recent studies have indicated a shift in the predominant pathogens of concern, with Streptococcus sp. a being superseded by Staphylococcus sp. and Enterococcus sp. as the leading causes of concern. This shift is of concern as it is associated with Staphylococcus Aureus which has a high virulence rate and a tendency to form a biofilm, meaning that non-surgical therapy may not be effective. It is imperative to deliberate on the likelihood of platelet blood clot formation, which may be accompanied by bacterial infestation and the development of a biofilm. Methods: MEDLINE, Embase, and Pubmed were searched using terms relating to ‘endocarditis’ and ‘Staphilococcus aureus’, along with ‘epidemiology’, ‘pathogenesis’, ‘coagulation’, ‘platelet’, ‘aggregation’, and ‘immunity’. The search focused on publications from the past 15 years, but excluded older, highly regarded articles. We also searched the reference lists of relevant articles. Recommended review articles are cited for more details. Results: An endocarditis lesion is believed to be a blood clot infected with bacteria that adheres to the heart valves. Infective endocarditis is a good example of immunothrombosis, where the coagulation system, innate immunity and the function of coagulation in isolating and eliminating pathogens interact. However, in the context of infective endocarditis, immunothrombosis unintentionally establishes an environment conducive to bacterial proliferation. The process of immunothrombosis impedes the immune system, enabling bacterial proliferation. The coagulation system plays a pivotal role in the progression of this condition. Conclusion: The coagulation system is key to how bacteria attach to the heart valves, how vegetations develop, and how complications like embolisation and valve dysfunction occur. Staphylococcus aureus, the main cause of infective endocarditis, can change blood clotting, growing well in the fibrin-rich environment of vegetation. The coagulation system is a good target for treating infective endocarditis because of its central role in the disease. But we must be careful, as using blood-thinning medicines in patients with endocarditis can often lead to an increased risk of bleeding. Full article

(This article belongs to the Section Lipid Metabolism)

► Show Figures

Graphical abstract

17 pages, 4465 KiB

Open AccessArticle

Metabolomic Analysis Revealed the Differences in Metabolites Between Three Different Sugarcane Stems and Leaves

by Hongbo Lou, Linyan Xie, Xianhong Wang, Xianli Li, Lilian He and Fusheng Li

Metabolites 2025, 15(5), 327; https://doi.org/10.3390/metabo15050327 - 15 May 2025

Viewed by 459

Abstract

Background: Sugarcane is an important sugar crop. Sugarcane stems are mainly used for sugar extraction, while leaves can only be burned as waste. However, sugarcane leaves can also produce a large number of secondary metabolites, and these metabolites have significant nutritional and pharmacological [...] Read more.

Background: Sugarcane is an important sugar crop. Sugarcane stems are mainly used for sugar extraction, while leaves can only be burned as waste. However, sugarcane leaves can also produce a large number of secondary metabolites, and these metabolites have significant nutritional and pharmacological value. At present, there are few studies on sugarcane compounds. Methods: Therefore, the stems and leaves of three sugarcane varieties (Yacheng 89-159, Dianzhe 01-58, ROC22) were selected as experimental materials, and the compounds of stems and leaves of different sugarcane were studied using high-performance liquid chromatography. Results: Metabolomics analysis detected 1197 metabolites that could be broadly divided into 11 categories. Orthogonal partial least squares discriminant analysis identified metabolites that were differentially abundant across groups (stems and leaves within and across the three varieties). Flavonoids, phenolic acids, and lipids were the main differential metabolites. Notably, tricin-4′-O-(guaiacylglycerol)ether-7-O-glucoside, quercetin-3,4-O-di-glucoside, cyanidin-3-O-(6′′-O-malony)glucoside were significantly higher in the stems than in the leaves across all three varieties. The content of methylenesuccinic acid was higher in the leaves of Dianzhe 01-58 and ROC22. In the comparative analysis of the top 20 differential metabolites among different varieties, it was found that the metabolite content of stems and leaves of Yacheng 89-9 and ROC22 was significantly higher than that of Dianzhe 01-58. Next, KEGG analysis showed that these differential metabolites were mainly enriched in pathways related to flavonoid, phenylpropanoid, and isoflavonoid biosynthesis, as well as starch and sucrose metabolism. Leaves also had significantly fewer metabolites involved in starch and sucrose metabolism than stems did. Conclusion: In conclusion, this study provides a scientific basis for utilization of sugarcane compounds, laying a theoretical foundation for further processing of sugarcane by-products into higher-value materials. Full article

(This article belongs to the Section Plant Metabolism)

► Show Figures

Figure 1

23 pages, 7907 KiB

Open AccessArticle

Exploring the Mechanism of Luteolin in Protecting Chickens from Ammonia Poisoning Based on Proteomic Technology

by Yu Jin, Azi Shama, Haojinming Tang, Ting Zhao, Xinyu Zhang, Falong Yang and Dechun Chen

Metabolites 2025, 15(5), 326; https://doi.org/10.3390/metabo15050326 - 14 May 2025

Viewed by 411

Abstract

Background: Ammonia (NH₃), a harmful gas, reduces livestock productivity, threatens their health, and causes economic losses. Luteolin (Lut), an anti-inflammatory flavonoid, may counteract these effects. Methods: Our study explored luteolin’s protective mechanisms on chicken splenic lymphocytes under ammonia stress using a [...] Read more.

Background: Ammonia (NH₃), a harmful gas, reduces livestock productivity, threatens their health, and causes economic losses. Luteolin (Lut), an anti-inflammatory flavonoid, may counteract these effects. Methods: Our study explored luteolin’s protective mechanisms on chicken splenic lymphocytes under ammonia stress using a simulation model and four-dimensional fast data-independent acquisition (4D-FastDIA) proteomics. We identified 316 proteins, with 69 related to ammonia’s negative effects and 247 to Lut’s protection. Thirty differentially expressed proteins (DEPs) were common to both groups, with 27 showing counter-regulation with Lut. Results: Gene Ontology (GO) analysis showed DEPs enriched in molecular responses to interferons and the negative regulation of immune responses, mainly located extracellularly. Molecular function analysis revealed DEPs in antigen binding and synthase activity. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis linked DEPs to pathways like estrogen signaling, NOD-like receptor signaling, cytokine–cytokine receptor interaction, and JAK-STAT signaling. The quantitative real-time polymerase chain reaction (qRT-PCR) results indicated that the mRNA levels of Interferon Alpha and Beta Receptor subunit 2 (IFNAR2) and Signal Transducer and Activator of Transcription 1 (STAT1) were trending downward. This observation was in strong agreement with the downregulation noted in the proteomics analysis. Conclusions: Lut’s protective role against ammonia’s adverse effects on chicken splenic lymphocytes is linked to the modulation of key signaling pathways, offering insights for further research on treating ammonia exposure with Lut. Full article

(This article belongs to the Special Issue Metabolomics Insights into Environmental Pollution: Unraveling Metabolic Responses to Contaminants)

► Show Figures

Graphical abstract

15 pages, 9026 KiB

Open AccessArticle

Integrated Analysis of Volatile Metabolites in Rose Varieties: Effects of Cultivar Differences and Drying Temperatures on Flavor Profiles

by Jun Zhang, Meile Sun, Xiangrong Ren, Jing Yang, Yijie Zhang, Jingtao Hui, Pengbing Li, Jianfei Tao, Tianzhi Liu and Guocang Lin

Metabolites 2025, 15(5), 325; https://doi.org/10.3390/metabo15050325 - 14 May 2025

Viewed by 413

Abstract

Background: Rose processing faces critical challenges in preserving bioactive compounds and aroma profiles during thermal treatments, particularly given the growing demand for natural ingredients in the food and cosmetic industries. Methods: Using widely targeted metabolomics, we first characterized volatile profiles of four major [...] Read more.

Background: Rose processing faces critical challenges in preserving bioactive compounds and aroma profiles during thermal treatments, particularly given the growing demand for natural ingredients in the food and cosmetic industries. Methods: Using widely targeted metabolomics, we first characterized volatile profiles of four major commercial cultivars (Hetian, Damask, Bulgarian, and Fenghua; n = 6 replicates per cultivar), identifying terpenoids as dominant components (p < 0.05). Subsequent thermal optimization focused on Hetian rose, where WGCNA and K-means analyses revealed temperature-dependent dynamics (40–55 °C, triplicate drying trials per temperature). Results: Hetian rose exhibited significantly higher accumulation (p < 0.05) of a unique sesquiterpene marker, 4-(1,5-dimethyl-1,4-hexadienyl)-1-methyl-cyclohexene. Systematic drying optimization identified 50 °C as the thermal threshold for optimal color, bioactive retention, and sensory quality. Mechanistic analysis identified 193 temperature-responsive metabolites (VIP > 1, FC < 0.25 or >4, p < 0.01), with terpenoid biosynthesis (MVA/MEP pathways) and esterification dynamics emerging as critical control points. Conclusions: This study establishes the first cultivar-specific processing framework for roses, demonstrating that metabolic signature-guided drying improves product quality. The findings advance our understanding of thermal impacts on aroma biochemistry while providing actionable protocols for natural product industries. Full article

(This article belongs to the Section Plant Metabolism)

► Show Figures

Figure 1

12 pages, 1831 KiB

Open AccessArticle

Intestinal Epithelial-Derived Exosomes Under Cold Stimulation Promote Adipose Thermogenesis

by Xue Han, Tiange Feng, Yaxu Yang, Ziming Zhu, Fangyu Shao, Lijun Sun, Yue Yin and Weizhen Zhang

Metabolites 2025, 15(5), 324; https://doi.org/10.3390/metabo15050324 - 14 May 2025

Viewed by 409

Abstract

Background: Whether intestinal epithelial cells can regulate distant adipose tissue remains a mystery. Methods: Cold-stimulated intestinal epithelial cell-derived exosomes (Cold IEC-Exo) play a pivotal role in enhancing adipose thermogenesis and metabolic homeostasis, as demonstrated in this study. Results: IEC-Exo can [...] Read more.

Background: Whether intestinal epithelial cells can regulate distant adipose tissue remains a mystery. Methods: Cold-stimulated intestinal epithelial cell-derived exosomes (Cold IEC-Exo) play a pivotal role in enhancing adipose thermogenesis and metabolic homeostasis, as demonstrated in this study. Results: IEC-Exo can accumulate in adipose tissue. Compared with IEC-Exo derived from room temperature mice (RT IEC-Exo), Cold IEC-Exo significantly enhanced the thermogenesis of adipose. In vitro, Cold IEC-Exo directly stimulated thermogenesis in primary adipocytes by elevating oxygen consumption rate, proton leak, and fatty acid uptake, with no effect on glucose uptake. Small RNA sequencing identified miR-674-3p as a key mediator enriched in Cold IEC-Exo. miR-674-3p mimicry replicated Cold IEC-Exo effects, augmenting Ucp1 expression, mitochondrial uncoupling, and fatty acid utilization in adipocytes. Local overexpression of miR-674-3p in BAT and sWAT via AAV in vivo enhanced thermogenesis and attenuated diet-induced glucose intolerance. Conclusions: These findings establish that Cold IEC-Exo, via miR-674-3p transfer, drive adipose thermogenic activation and mitigate metabolic dysfunction, highlighting their therapeutic potential in obesity-related disorders. Full article

(This article belongs to the Special Issue Energy Metabolism in Brown Adipose Tissue)

► Show Figures

Figure 1

15 pages, 1440 KiB

Open AccessArticle

Plasma Metabolic Outliers Identified in Estonian Human Knockouts

by Ketian Yu, Estonian Biobank Research Team, Karol Estrada, Tõnu Esko, Mart Kals, Tiit Nikopensius, Jaanika Kronberg, Urmo Võsa, Arthur Wuster and Lorenzo Bomba

Metabolites 2025, 15(5), 323; https://doi.org/10.3390/metabo15050323 - 13 May 2025

Viewed by 438

Abstract

Background/Objectives: Metabolomics, in combination with genetic data, is a powerful approach to study the biochemical consequences of genetic variation. We assessed the impact of human gene knockouts (KOs) on the metabolite levels of Estonia Biobank (EstBB) participants and integrated the results with electronic [...] Read more.

Background/Objectives: Metabolomics, in combination with genetic data, is a powerful approach to study the biochemical consequences of genetic variation. We assessed the impact of human gene knockouts (KOs) on the metabolite levels of Estonia Biobank (EstBB) participants and integrated the results with electronic health record data. Methods: In 150,000 EstBB genotyped participants, we identified 723 KOs with 152 different predicted loss of function (pLoF) variants in 115 genes. For those KOs and 258 controls, 1387 metabolites were profiled using ultra-high-performance liquid chromatography–tandem mass spectrometry. Results: We identified 48 associations linking rare pLoF variants in 22 genes to 43 metabolites. Out of 48 associations, 27 (56%) were found in genes that cause inborn errors of metabolism. The top associations identified in our analysis included genes and metabolites involved in the degradation pathway of the pyrimidine bases uracil and thymine (DPYD and UPB1). We found DPYD gene KOs to be associated with elevated levels of Uracil, confirming that DPD-deficiency is a leading cause of severe 5-Fluorouracil toxicity. Overall, 54% of reported associations are gene targets of approved drugs or bioactive drug-like compounds. Conclusions: Our findings contribute to assessing the impact of human KOs on metabolite levels and offer insights into gene functions, disease mechanism, and drug target validation. Full article

(This article belongs to the Section Bioinformatics and Data Analysis)

► Show Figures

Figure 1

15 pages, 2494 KiB

Open AccessArticle

The Effect of Dexamethasone-Mediated Atrophy on Mitochondrial Function and BCAA Metabolism During Insulin Resistance in C2C12 Myotubes

by Kayla J. Ragland, Kipton B. Travis, Emmalie R. Spry, Toheed Zaman, Pamela M. Lundin and Roger A. Vaughan

Metabolites 2025, 15(5), 322; https://doi.org/10.3390/metabo15050322 - 13 May 2025

Viewed by 578

Abstract

Background: Muscle loss during sarcopenia and atrophy is also commonly associated with age-related insulin resistance. Interestingly, branched-chain amino acids (BCAA) which are known for stimulating muscle protein synthesis are commonly elevated during insulin resistance and sarcopenic obesity. Objectives: This study investigated [...] Read more.

Background: Muscle loss during sarcopenia and atrophy is also commonly associated with age-related insulin resistance. Interestingly, branched-chain amino acids (BCAA) which are known for stimulating muscle protein synthesis are commonly elevated during insulin resistance and sarcopenic obesity. Objectives: This study investigated the effects of the interplay between atrophy and insulin resistance on insulin sensitivity, mitochondrial metabolism, and BCAA catabolic capacity in a myotube model of skeletal muscle insulin resistance. Methods: C2C12 myotubes were treated with dexamethasone to induce atrophy. Insulin resistance was induced via hyperinsulinemia. Gene and expression were measured using qRT-PCR and Western blot, while mitochondrial and lipid content were assessed using fluorescent staining. Cell metabolism was analyzed via Seahorse metabolic assays. Results: Both dexamethasone-induced atrophy and insulin resistance independently reduced insulin-stimulated pAkt levels, as well as mitochondrial function and content. However, neither treatment affected gene or protein expression associated with mitochondrial biogenesis or content. Although dexamethasone independently reduced insulin sensitivity in otherwise previously insulin-sensitive cells, dexamethasone had no significant effect on extracellular BCAA content. Conclusions: Our findings indicate the metabolic interplay between atrophy and insulin resistance and demonstrate that both can reduce mitochondrial function, though only limited effects were observed on indicators of BCAA catabolism and utilization. This emphasizes the need for future studies to investigate the mechanisms that underlie atrophy and other metabolic disorders to develop new interventions. Full article

(This article belongs to the Section Endocrinology and Clinical Metabolic Research)

► Show Figures

Figure 1

27 pages, 2350 KiB

Open AccessArticle

Exploring Postharvest Metabolic Shifts and NOX2 Inhibitory Potential in Strawberry Fruits and Leaves via Untargeted LC-MS/MS and Chemometric Analysis

by Georgia Ladika, Paris Christodoulou, Eftichia Kritsi, Thalia Tsiaka, Georgios Sotiroudis, Dionisis Cavouras and Vassilia J. Sinanoglou

Metabolites 2025, 15(5), 321; https://doi.org/10.3390/metabo15050321 - 13 May 2025

Viewed by 391

Abstract

Background/Objectives: Strawberries are highly appreciated for their rich phytochemical composition, but rapid postharvest deterioration limits their shelf life and nutritional quality. This study aimed to investigate the metabolic changes occurring in both strawberry fruits and leaves during storage and to evaluate the NADPH [...] Read more.

Background/Objectives: Strawberries are highly appreciated for their rich phytochemical composition, but rapid postharvest deterioration limits their shelf life and nutritional quality. This study aimed to investigate the metabolic changes occurring in both strawberry fruits and leaves during storage and to evaluate the NADPH oxidase 2 (NOX2) inhibitory potential of strawberry-derived metabolites. Methods: Untargeted LC-MS/MS analysis was conducted on fruit and leaf tissues stored at 8 ± 0.5 °C. A total of 37 metabolites were identified, including organic acids, phenolic acids, flavonoids, and hydroxycinnamic acid derivatives. Multivariate statistical analyses (ANOVA, PLS-DA, and volcano plots) were used to assess temporal and tissue-specific metabolic shifts. Additionally, a machine learning-based predictive model was applied to evaluate the NOX2 inhibitory potential of 24 structurally characterized metabolites. Results: Storage induced significant and tissue-specific metabolic changes. In fruits, malic acid, caffeic acid, and quercetin-3-glucuronide showed notable variations, while ellagic acid aglycone and galloylquinic acid emerged as prominent markers in leaves. The predictive model identified 21 out of 24 metabolites as likely NOX2 inhibitors, suggesting potential antioxidant and anti-inflammatory bioactivity. Conclusions: These findings provide new insights into postharvest biochemical dynamics in both strawberry fruits and leaves. The results highlight the value of leaves as a source of bioactive compounds and support their potential valorization in functional food and nutraceutical applications. Full article

(This article belongs to the Special Issue Metabolic Changes during Pre- and Post-harvest Fruit and Vegetable Decay, Ripening and Senescence)

► Show Figures

Figure 1

12 pages, 636 KiB

Open AccessArticle

Longitudinal Trajectory of Free Fatty Acids in Pregnancy According to First-Trimester Maternal Metabolic Status and the Presence of Gestational Diabetes

by Otilia Perichart-Perera, Isabel González-Ludlow, Omar Piña-Ramírez, Maricruz Tolentino-Dolores, Guadalupe Estrada-Gutierrez, Sandra B. Parra-Hernández, Maribel Sánchez-Martínez, Omar Granados-Portillo and Ameyalli M. Rodríguez-Cano

Metabolites 2025, 15(5), 320; https://doi.org/10.3390/metabo15050320 - 11 May 2025

Viewed by 541

Abstract

Background/Objectives: Maternal free fatty acids (FFAs) play a critical role in maternal metabolism, fetal growth, and pregnancy outcomes. However, their relationship with maternal metabolic status in early pregnancy and the subsequent development of gestational diabetes mellitus (GDM) remains unclear. Aim: Assess the trajectory [...] Read more.

Background/Objectives: Maternal free fatty acids (FFAs) play a critical role in maternal metabolism, fetal growth, and pregnancy outcomes. However, their relationship with maternal metabolic status in early pregnancy and the subsequent development of gestational diabetes mellitus (GDM) remains unclear. Aim: Assess the trajectory of FFA concentrations during pregnancy, considering first-trimester metabolic status (obesity, insulin resistance—IR) and the development of GDM, and evaluate whether first-trimester FFA is a relevant risk factor for GDM. Methods: A case–control study nested within the OBESO cohort (Mexico City, pregnant women and their children), classified women according to first-trimester metabolic status (pregestational body mass index—pBMI, insulin resistance homeostasis model assessment—HOMA-IR > 1.6), as well as the presence of GDM: Group 1 (normal weight without IR, n = 60), Group 2 (obesity without IR, no GDM, n = 20), Group 3 (obesity with IR, no GDM, n = 20), and Group 4 (obesity with IR, with GDM, n = 9). FFA concentrations were measured each trimester. Statistical analyses included repeated measures ANOVA and logistic regression models. Results: FFA concentrations were the highest in Group 4 across all trimesters (p < 0.05). FFAs decreased throughout pregnancy in all groups (p = 0.023), with the most significant decline from the first to the third trimester (p < 0.001). The greatest reduction occurred in Group 4 (p < 0.001), followed by Group 3. Multivariate logistic regression showed no association between first-trimester FFAs and the development of GDM. Higher gestational weight gain was associated with a higher GDM risk (OR: 1.22, 95%CI: 1.01–1.48), when the FFAs difference was accounted for. Conclusions: FFA levels are higher in women with GDM compared with women with obesity or a normal weight. However, FFAs progressively decline from the first to the third trimester, with the most pronounced decrease in women with obesity, IR, and GDM. Full article

(This article belongs to the Special Issue Adipose Tissue, Reproduction and Metabolic Health in Women)

► Show Figures

Figure 1

14 pages, 3108 KiB

Open AccessArticle

Comprehensive Analysis of Small RNA Modifications in Arabidopsis thaliana and Their Dynamics During Seed Germination

by Liu-Cheng Jiang, Meng Men, Xuan-Jun Cui, Ren-Jie Zeng, Shu-Yi Gu, Tian Feng, Chen Zeng, Tiantian Ye, Jun Xiong, Bi-Feng Yuan and Yu-Qi Feng

Metabolites 2025, 15(5), 319; https://doi.org/10.3390/metabo15050319 - 10 May 2025

Viewed by 391

Abstract

Background: Small RNA, defined as RNA molecules of less than 200 nucleotides in length, play pivotal regulatory roles in plant growth, development, and environmental stress responses. However, research on modifications in plant small RNA remains limited. Methods: In this study, we [...] Read more.

Background: Small RNA, defined as RNA molecules of less than 200 nucleotides in length, play pivotal regulatory roles in plant growth, development, and environmental stress responses. However, research on modifications in plant small RNA remains limited. Methods: In this study, we developed a liquid chromatography–tandem mass spectrometry (LC-MS/MS) method for the simultaneous detection of 41 RNA modifications, facilitating the systematic qualification and quantification of modifications in plant small RNA. Results: We identified a total of nine modifications, among which N⁶,N⁶-dimethyladenosine (m^6,6A) is a newly identified modification in plant small RNA. Furthermore, we conducted a quantitative analysis of these modifications in Arabidopsis thaliana during the germination process and observed significant dynamic changes in their abundance from 1 to 5 days post-germination. Notably, the trends in the contents of these modifications exhibited a strong correlation with the reported gene expression levels of the relevant modifying enzymes and demodifying enzymes, suggesting that these modifications may play essential roles during seed germination and are tightly regulated by the genes of the corresponding enzymes. Conclusions: The discovery of these modifications in plant small RNA, coupled with the dynamic changes in their levels during germination, holds great promise for a further understanding of the physiological functions of small RNA modifications and their associated regulatory mechanisms in plant seed germination. Full article

(This article belongs to the Special Issue LC-MS/MS Analysis for Plant Secondary Metabolites)

► Show Figures

Graphical abstract

16 pages, 262 KiB

Open AccessArticle

Fatty Liver and Hyperuricemia in Workers: Combined Effects on Metabolic Dysfunction and the Role of Lifestyle Factors

by Jui-Hua Huang, Ren-Hau Li, Hon-Ke Sia and Feng-Cheng Tang

Metabolites 2025, 15(5), 318; https://doi.org/10.3390/metabo15050318 - 9 May 2025

Viewed by 449

Abstract

Background/Objectives: Fatty liver and hyperuricemia are growing public health concerns linked to unhealthy lifestyles, yet their combined effects in working populations remain underexplored. This study investigates their associations with metabolic risk factors, inflammation, and liver dysfunction to inform workplace health strategies. Methods [...] Read more.

Background/Objectives: Fatty liver and hyperuricemia are growing public health concerns linked to unhealthy lifestyles, yet their combined effects in working populations remain underexplored. This study investigates their associations with metabolic risk factors, inflammation, and liver dysfunction to inform workplace health strategies. Methods: The participants were employees aged 20 or older from four industrial enterprises located in central Taiwan. A total of 3089 participants (2571 males, 518 females) were analyzed. Lifestyle factors were assessed via a self-administered questionnaire, fatty liver was diagnosed using ultrasound, and serum uric acid levels, metabolic parameters, inflammatory markers, and liver function were measured. Results: The prevalence of fatty liver (43.2%) exceeded that of hyperuricemia (25.5%), with a higher burden among males. Fatty liver was associated with lower physical activity, while alcohol consumption was significantly higher in individuals with both conditions. Both conditions correlated with increased metabolic risk factors, liver dysfunction, and inflammation. Health-related risk factors were compared across four groups, using Group A (no hyperuricemia/no fatty liver, OR = 1.00) as the reference. The risk of metabolic syndrome increased progressively: 2.90 (Group B: hyperuricemia/no fatty liver), 6.15 (Group C: no hyperuricemia/fatty liver), and 11.52 (Group D: hyperuricemia/fatty liver), following the trend A < B < C < D. Notably, Group D had the highest risk, with exacerbated inflammation and liver dysfunction. Conclusions: Fatty liver and hyperuricemia synergistically worsen metabolic disorders, inflammation, and liver dysfunction. Early detection and lifestyle interventions are crucial to mitigating long-term health risks. Full article

(This article belongs to the Special Issue Liver Injury and Regeneration—Metabolic Research)

20 pages, 3404 KiB

Open AccessArticle

A Data-Driven Approach to Link GC-MS and LC-MS with Sensory Attributes of Chicken Bouillon with Added Yeast-Derived Flavor Products in a Combined Prediction Model

by Simon Leygeber, Carmen Diez-Simon, Justus L. Großmann, Anne-Charlotte Dubbelman, Amy C. Harms, Johan A. Westerhuis, Doris M. Jacobs, Peter W. Lindenburg, Margriet M. W. B. Hendriks, Brenda C. H. Ammerlaan, Marco A. van den Berg, Rudi van Doorn, Roland Mumm, Age K. Smilde, Robert D. Hall and Thomas Hankemeier

Metabolites 2025, 15(5), 317; https://doi.org/10.3390/metabo15050317 - 8 May 2025

Viewed by 653

Abstract

Background: There is a continuous demand to create new, superior sensory food experiences. In the food industry, yeast-derived flavor products (YPs) are often used as ingredients in foods to create new aromas and taste qualities that are appreciated by consumers. Methods: Chicken bouillon [...] Read more.

Background: There is a continuous demand to create new, superior sensory food experiences. In the food industry, yeast-derived flavor products (YPs) are often used as ingredients in foods to create new aromas and taste qualities that are appreciated by consumers. Methods: Chicken bouillon samples containing diverse YPs were chemically and sensorially characterized using statistical multivariate analyses. The sensory evaluation was performed using quantitative descriptive analysis (QDA) by trained panelists. Thirty-four sensory attributes were scored, including odor, flavor, mouthfeel, aftertaste and afterfeel. Untargeted metabolomic profiles were obtained using stir bar sorptive extraction (SBSE) coupled to GC-MS, RPLC-MS and targeted HILIC-MS. Results: In total, 261 volatiles were detected using GC-MS, from chemical groups of predominantly aldehydes, esters, pyrazines and ketones. Random Forest (RF) modeling revealed volatiles associated with roast odor (2-ethyl-5-methyl pyrazine, 2,3,5-trimethyl-6-isopentyl pyrazine) and chicken odor (2,4-nonadienal, 2,4-decadienal, 2-acetyl furan), which could be predicted by our combined model with R² > 0.5. In total, 2305 non-volatiles were detected for RPLC-MS and 34 for targeted HILIC-MS, where fructose-isoleucine and cyclo-leucine-proline were found to correlate with roast flavor and odor. Furthermore, a list of metabolites (glutamate, monophosphates, methionyl-leucine) was linked to umami-related flavor. This study describes a straightforward data-driven approach for studying foods with added YPs to identify flavor-impacting correlations between molecular composition and sensory perception. It also highlights limitations and preconditions for good prediction models. Overall, this study emphasizes a matrix-based approach for the prediction of food taste, which can be used to analyze foods for targeted flavor design or quality control. Full article

(This article belongs to the Section Food Metabolomics)

► Show Figures

Figure 1

16 pages, 4798 KiB

Open AccessArticle

Effects of Maillard Reaction Products on Skeletal Muscle Cells: An In Vitro Study Using C2C12 Myotubes

by Marina Miyaki, Yusuke Komiya, Itsuki Sumiya, Rina Yamaguchi, Moeka Kuno, Chika Kojima, Ryosuke Makino, Takahiro Suzuki, Yoshihiro Suzuki, Issei Yokoyama and Keizo Arihara

Metabolites 2025, 15(5), 316; https://doi.org/10.3390/metabo15050316 - 8 May 2025

Viewed by 671

Abstract

Background: Maillard reaction products (MRPs) are known for their antioxidant properties; however, their effects on muscle cells remain unclear. This study aims to elucidate the effects of MRPs on muscle hypertrophy and atrophy in C2C12 myotubes. Methods: MRPs were prepared by [...] Read more.

Background: Maillard reaction products (MRPs) are known for their antioxidant properties; however, their effects on muscle cells remain unclear. This study aims to elucidate the effects of MRPs on muscle hypertrophy and atrophy in C2C12 myotubes. Methods: MRPs were prepared by heating L-lysine and D-glucose, and their antioxidant activity was assessed using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay. Subsequently, mouse C2C12 myoblasts were cultured with MRPs until myotubes formed, and their diameters were measured to assess hypertrophic and atrophic changes. Akt phosphorylation was evaluated by Western blotting, and gene expression levels were analyzed via quantitative PCR. Results: The prepared MRPs exhibited high antioxidant activity in the DPPH radical scavenging assay. MRP treatment significantly increased the average myotube diameter by approximately 40% and enlarged the largest myotube diameter by up to 80%, potentially mediated by enhanced Akt phosphorylation. Under dexamethasone-induced atrophy, MRPs modestly attenuated the reduction in myotube diameter by approximately 20%, although the effect was not statistically significant, and did not significantly alter the fusion index either. Quantitative PCR analysis revealed that MRP treatment significantly reduced the mRNA expression of Nfe2l2, a key regulator of antioxidant response, whereas it had no notable effects on the expression of genes related to myoblast proliferation (Myod1), differentiation (Myog), hypertrophy (Igf1), atrophy (Foxo1 and Trim63), and oxidative stress (Cat, Gclc, and Nqo1). Conclusions: Our findings suggested that MRPs possess antioxidant activity and promote myotube hypertrophy via Akt signaling. This study highlighted the potential of MRPs as functional ingredients for promoting muscle health, though further in vivo studies are required to validate their physiological relevance. Full article

(This article belongs to the Special Issue Muscle Metabolic Response and Adaptation to Exercise, Diet, and Environment)

► Show Figures