Metabolites

25 pages, 3659 KB

Open AccessEditor’s ChoiceReview

Mangiferin as a Multilevel Modulator of Metabolic Syndrome: Current Evidence and Future Perspectives

by Joan A. Ramos-Olvera, J. Martín Torres-Valencia, Mario I. Ortiz, Eduardo Fernández-Martínez, Carlos A. Gómez-Aldapa, Víctor Manuel Muñoz-Pérez and Raquel Cariño-Cortés

Metabolites 2026, 16(7), 453; https://doi.org/10.3390/metabo16070453 - 27 Jun 2026

Viewed by 630

Abstract

Background/Objectives: The obesogenic environment is a key factor in the rising prevalence of metabolic syndrome (MetS), a major global public health challenge. Mangiferin (Mgf) is a C-glycosylated xanthone, and its preventive and therapeutic potential stems from its multisystemic pharmacological profile. This review [...] Read more.

Background/Objectives: The obesogenic environment is a key factor in the rising prevalence of metabolic syndrome (MetS), a major global public health challenge. Mangiferin (Mgf) is a C-glycosylated xanthone, and its preventive and therapeutic potential stems from its multisystemic pharmacological profile. This review integrates findings on Mgf in the management of metabolic syndrome, aiming to identify gaps and propose prospective studies to enable the scaling up of Mgf for clinical applications. Methods: To this end, a literature search was conducted, the level of evidence was identified, and the available scientific information on Mgf in metabolic syndrome was synthesized from PubMed, Scopus, and Web of Science from 2016 to 2025. Results: Mgf improves overall metabolic dysfunction by activating the AMPK pathway, reduces inflammation by activating Nrf2, suppressing NF-κB, and decreasing pro-inflammatory mediators (COX-2, IL-6, NLRP3), modulates CB1 and PPAR receptors and other markers associated with obesity (adiponectin, leptin, resistin), as well as autophagy processes. However, most of the evidence comes from in silico, in vitro, and preclinical studies, with few clinical observations. Conclusions: This underscores the need to integrate studies on the correlation between the bioavailability of Mgf and norathyriol with the regulation of the microbiota, as well as their effects on metabolomic and epigenetic mechanisms in MetS. Full article

(This article belongs to the Section Plant Metabolism)

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26 pages, 7002 KB

Open AccessEditor’s ChoiceArticle

Proteomics and Metabolomics Reveal Novel Impacts of Choline Supply on Calf Hepatocytes Experiencing Accumulation During a Fatty Acid Challenge

by Yaqi Chang, Bin Jia, Yaran Si, Zexin Zhang, Jiachen Liu, Yue Gao, Junhao Wang, Yanhui Wang, Juan J. Loor, Bingbing Zhang and Wei Yang

Metabolites 2026, 16(7), 451; https://doi.org/10.3390/metabo16070451 - 26 Jun 2026

Viewed by 561

Abstract

Background/Objectives: Exposure to high and sustained levels of non-esterified fatty acids (NEFA) in the peripartal period is the main cause of fatty liver disease in dairy cows. Rumen-protected choline is often fed as part of the nutritional management of peripartal cows, with in [...] Read more.

Background/Objectives: Exposure to high and sustained levels of non-esterified fatty acids (NEFA) in the peripartal period is the main cause of fatty liver disease in dairy cows. Rumen-protected choline is often fed as part of the nutritional management of peripartal cows, with in vivo and in vitro data indicating positive effects of this nutrient on alleviating liver lipid accumulation. Although hepatic molecular mechanisms associated with choline supply have been studied using a target gene, protein, or metabolite approach, application of high-throughput technologies could vastly enhance fundamental knowledge on the functional role of choline. The main objective was to challenge isolated hepatocytes with a mixture of NEFA and determine proteome- and metabolome-wide effects in response to choline supply. Methods: Three healthy female calves (1 d old, 30–45 kg) were sacrificed to harvest hepatocytes. During a 12 h incubation, isolated hepatocytes were challenged without NEFA (control), 1.2 mM NEFA (c9-18:1, 18:2, 16:0, 18:0, and c9-16:1 at 43.5%, 4.9%, 31.9%, 14.4%, and 5.3% of total NEFA, respectively), or NEFA for 6 h followed by 10 μM choline chloride for another 6 h (NEFA + Chol). iTRAQ labeling-based protein profiling and GC/MS-based metabolomics profiling were used to determine changes in proteins and metabolites. Differentially abundant proteins for each group comparison were determined at a threshold of 1.4-fold change. Differences in metabolite profiles were assessed via pairwise comparisons. A subset of differentially abundant proteins was validated via qRT-PCR and Western blotting. Results: Compared with the control, there were 90 proteins and 22 metabolites in the NEFA group, and 83 proteins and 29 metabolites in the NEFA + Chol. Compared with NEFA, there were 49 proteins and 17 metabolites in the NEFA + Chol group. Greater abundance of hexokinase-1 (HK1), fructose-bisphosphate aldolase (ALDOA), mitochondrial pyruvate carrier 1 (MPC1), and increased concentrations of lactate with high NEFA treatment alone suggested greater glycolytic and TCA cycle activity. Accumulation of triacylglycerol in the NEFA group was associated with lipotoxicity and markers of inflammation, such as greater abundance of prostaglandin reductase 1 (PTGR1), serious cell autophagy processes, such as greater abundance of cell division cycle 42 (CDC42), and NFκB-related proteins. Choline supplementation reduced TAG partly due to greater VLDL secretion driven by greater abundance of diacylglycerol acyltransferase (DGAT1), perilipin 3 (PLIN3), and apolipoprotein C-III (APOC3). In addition, a greater abundance of carnitine O-palmitoyltransferase 1b (CPT1B) with choline suggested enhanced mitochondrial β-oxidation. Activation of the CDC42/JNK pathway and ROS/NFκB axis-related proteins, along with depressed PI3K/AKT/RAC-related proteins, indicated enhanced mitochondrial autophagy in response to NEFA. Conclusions: Overall, data confirmed published effects of choline on TAG accumulation, VLDL secretion, and fatty acid oxidation, while highlighting negative effects of NEFA on the respiratory electron transport chain, autophagy, and inflammatory processes. Full article

(This article belongs to the Special Issue Metabolic Research in Dairy Cattle Health)

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13 pages, 6810 KB

Open AccessEditor’s ChoiceArticle

Reduced Histidine Metabolism Is Associated with Early Allograft Dysfunction Following Liver Transplantation

by Alissa M. Cutrone, Thomas Agius, Sofia Baptista, Eleonore Baughan, Korkut Uygun, Alban Longchamp and Heidi Yeh

Metabolites 2026, 16(7), 449; https://doi.org/10.3390/metabo16070449 - 26 Jun 2026

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Abstract

Background/Objectives: Early allograft dysfunction (EAD) is a common complication after liver transplantation and is associated with inferior graft survival. While normothermic machine perfusion (NMP) has reduced EAD incidence, the prediction of early graft performance prior to implantation remains elusive. We aimed to [...] Read more.

Background/Objectives: Early allograft dysfunction (EAD) is a common complication after liver transplantation and is associated with inferior graft survival. While normothermic machine perfusion (NMP) has reduced EAD incidence, the prediction of early graft performance prior to implantation remains elusive. We aimed to correlate the peri-transplant energetic and metabolic profile of liver grafts with post-transplant outcome in a cohort that included grafts preserved with NMP. Methods: Sequential biopsies were taken from 20 transplanted livers (10 immediate graft function [IGF] and 10 EAD), preserved by either static cold storage or NMP. Samples were collected immediately prior to implantation and 30 min after hepatic arterial reperfusion. Untargeted liquid chromatography-mass spectrometry was performed, and energy charge was calculated as (ATP + 1/2 ADP)/(ATP + ADP + AMP). Univariate and receiver operating characteristic analysis identified metabolites correlated with EAD and assessed predictive accuracy. Results: Hepatic concentrations of adenine nucleotides and calculated energy charge did not differ between outcome groups either before implantation or after reperfusion. In contrast, trans-urocanate was significantly enriched in IGF livers across both time points, and additional histidine catabolism pathway metabolites were preferentially increased in IGF grafts. Trans-urocanate demonstrated discriminatory performance for EAD with 80% sensitivity and 80% specificity, confirmed as the single strongest predictive feature among >1600 detected metabolites. Conclusions: These data identify histidine catabolism as a novel metabolic pathway associated with early graft function and a potential indicator of allograft resilience to ischemia-reperfusion injury. Integration of histidine pathway metabolites into perfusion-era viability assessment may serve as a discriminative biomarker of EAD and support future metabolite-guided graft optimization strategies. Full article

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

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18 pages, 860 KB

Open AccessEditor’s ChoiceArticle

Differential Effects of Turmeric Bioactive Compounds on Neuroinflammation and Mitochondrial Homeostasis in Brain Regions in a Rodent Model of Neuropathic Pain

by Xiaobo Liu, Julianna M. Santos, Takaki Kiritoshi, Guangchen Ji, Volker Neugebauer and Chwan-Li Shen

Metabolites 2026, 16(7), 442; https://doi.org/10.3390/metabo16070442 - 25 Jun 2026

Viewed by 447

Abstract

Background: Managing neuropathic pain (NP) is particularly challenging in the context of opioid use, and the mechanisms behind chronic pain remain unclear. Objective: This study evaluated the impact of turmeric bioactive compounds on brain regions including frontal cortex (FC), hippocampus (HPC), and hypothalamus [...] Read more.

Background: Managing neuropathic pain (NP) is particularly challenging in the context of opioid use, and the mechanisms behind chronic pain remain unclear. Objective: This study evaluated the impact of turmeric bioactive compounds on brain regions including frontal cortex (FC), hippocampus (HPC), and hypothalamus (HPT) in the spinal nerve ligation (SNL) in a rat model of NP. Methods: Twenty-four SD rats were assigned to four groups (N = 6 per group), namely sham+vehicle (Sham-V), SNL+vehicle (SNL-V), SNL + 100 mg/kg curcumin (SNL+100CUR), and SNL + 50 mg/kg bisdemethoxycurcumin (SNL+50BDMC), treated daily for four weeks via oral gavage. Gene expression levels related to neuroinflammation, oxidative stress, and mitochondrial homeostasis were measured using qRT-PCR. Protein-level or functional mitochondrial assays were not performed due to limited sample availability. Results: In the FC, SNL decreased the expression level of NRF1 and OPA1, but only OPA1 was increased by BDMC. In the HPC, SNL increased CD11b, NRF2, and MFN1; BDMC decreased CD11b and increased IBA1, NRF1, TFAM, PGC1α and Complex I; and CUR increased NRF1, TFAM, DRP1 and Complex I levels. In the HPT, SNL decreased GFAP and MFN1, with CUR and BDMC further decreasing GFAP but not affecting MFN1. Additionally, CUR and BDMC decreased the expression of several key markers of neuroimmune signaling and mitochondrial homeostasis, including IBA1, CD11b, NFkB, NRF1/2, DRP1, OPA1, PGC1α, TFAM, and PINK1. Conclusions: CUR and BDMC induced region-specific transcriptional remodeling of mitochondrial homeostasis across FC, HPC, and HPT in SNL rats, with somewhat limited effects in the FC, mixed effects in the HPC, and broader downregulation in the HPT. Full article

(This article belongs to the Special Issue Effects of Secondary Plant Metabolites on Human Health)

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20 pages, 5553 KB

Open AccessEditor’s ChoiceReview

Insulin Clearance Along the Liver–Kidney Axis: Implications for Insulin Action

by Germán Perdomo, Irene Cózar-Castellano and Sonia M. Najjar

Metabolites 2026, 16(7), 439; https://doi.org/10.3390/metabo16070439 - 24 Jun 2026

Viewed by 495

Abstract

The pleiotropic actions of insulin are mediated by cascades of signaling pathways and are regulated by circulating insulin levels. Under physiologic conditions, insulin levels reflect the balance between pancreatic beta-cell secretion and insulin clearance, which occurs primarily in liver hepatocytes and, to a [...] Read more.

The pleiotropic actions of insulin are mediated by cascades of signaling pathways and are regulated by circulating insulin levels. Under physiologic conditions, insulin levels reflect the balance between pancreatic beta-cell secretion and insulin clearance, which occurs primarily in liver hepatocytes and, to a lesser extent, in kidney proximal tubule cells. Therefore, coordination between insulin secretion and clearance is essential for systemic insulin sensitivity. Whereas insulin secretion is widely investigated, exploring the role of insulin clearance in regulating insulin sensitivity remains limited. This review summarizes the main mechanisms underlying insulin clearance along the liver–kidney axis and discusses how they contribute to metabolic regulation in health and disease. Full article

(This article belongs to the Special Issue Insulin Clearance and Metabolic Dysregulation in Health and Disease)

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22 pages, 2940 KB

Open AccessEditor’s ChoiceArticle

Monitoring Atypical Metabolite Biomarkers in Patients with Bile Acid Synthesis Disorders by a Novel Targeted Tandem Mass Spectrometry Assay

by Kenneth D. R. Setchell, Xueheng Zhao, Stacey Reed and Wujuan Zhang

Metabolites 2026, 16(7), 436; https://doi.org/10.3390/metabo16070436 - 23 Jun 2026

Viewed by 517

Abstract

Background/Objectives: Bile acid synthesis disorders (BASDs) represent a distinct category of progressive familiar cholestatic liver disease. A novel targeted mass spectrometry assay was developed for the accurate measurement of the major urinary atypical bile acids and bile alcohols that are biomarkers for [...] Read more.

Background/Objectives: Bile acid synthesis disorders (BASDs) represent a distinct category of progressive familiar cholestatic liver disease. A novel targeted mass spectrometry assay was developed for the accurate measurement of the major urinary atypical bile acids and bile alcohols that are biomarkers for HSD3B7, AKR1D1, CYP7B1 and CYP27A1 deficiencies, the four most common BASDs. Methods: Stable-isotope dilution UPLC tandem mass spectrometry was used for the simultaneous quantification of 12 key atypical bile acid biomarkers in urine from patients with BASD. Typical concentration ranges for these metabolites were established from urine samples from patients with biochemically and/or genetically confirmed BASD and compared with non-cholestatic and cholestatic controls. Results: The separation of major 3β-hydroxy-Δ⁵-bile acid sulfates, taurine- and glycine-conjugated 3-oxo-Δ⁴-bile acids, and bile alcohol glucuronides was achieved in a 20 min chromatographic run with intra- and inter-batch imprecisions of <15% for all metabolites. The mean ± SEM urinary concentration of total 3β-sulfated-Δ⁵-cholenoic acids in patients with HSD3B7 deficiency was 704 ± 204 µmol/L (n = 22), approximately 2000-fold higher than in cholestastic patients (n = 168) or non-cholestatic controls (n = 127). Similarly, the concentration of 5β-cholestane-3α,7α,12α,24,25-pentol-glucuronide, the major bile alcohol, in patients with CYP27A1 deficiency was 95 ± 17 µmol/L (n = 12). For CYP7B1 deficiency, two confirmed cases showed elevated levels (average, 7.5 µmol/L) of the glycine conjugate of 3β-sulfooxy-Δ⁵-bile acid. In AKR1D1 deficiency, total 3-oxo-Δ⁴-bile acids in urine were elevated (81 ± 16 µmol/L, n = 48), but concentrations showed overlap with cholestatic and non-cholestatic controls. Conclusions: A novel quantitative tandem mass spectrometry assay is described for the measurement of the major atypical metabolites and biomarkers in urine applicable to the accurate monitoring of treatment responses, and for the first time typical concentration ranges are established for each of these BASDs. Full article

(This article belongs to the Special Issue The Role of Lipid Metabolism in Health and Disease)

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19 pages, 1936 KB

Open AccessEditor’s ChoiceReview

The Gut Microbiome in Heart Failure: Pathways to Inflammation and Therapeutic Targets

by Uday Sankar Akash Vankayala, Ali Sohail, Bivin George, Madhu Singh, Omar Khayat, Malek Kreidieh, Alia Hasham and Luis Quiel

Metabolites 2026, 16(6), 431; https://doi.org/10.3390/metabo16060431 - 19 Jun 2026

Cited by 1 | Viewed by 889

Abstract

Heart failure (HF) continues to be a major global health burden, with persistent morbidity and mortality despite guideline-directed and device-based therapies. Evidence suggests the gut–heart axis is a critical and underrecognized contributor to HF progression. Alterations in cardiac output and systemic venous congestion [...] Read more.

Heart failure (HF) continues to be a major global health burden, with persistent morbidity and mortality despite guideline-directed and device-based therapies. Evidence suggests the gut–heart axis is a critical and underrecognized contributor to HF progression. Alterations in cardiac output and systemic venous congestion in HF lead to intestinal hypoperfusion, mucosal edema, and loss of barrier integrity, increasing intestinal permeability, gut dysbiosis, and translocation of microbial products. This systemic translocation is associated with chronic low-grade inflammation that activates innate immune pathways that correlate with endothelial dysfunction, oxidative stress, fibroblast activation, and adverse cardiac remodeling. Gut-derived metabolites derived by microbial metabolism modulate cardiovascular health by altering the metabolic profiles. Dysbiosis results in loss of protective short-chain fatty acid (SCFA)-producing bacteria and enriches pro-inflammatory taxa such as trimethylamine N-oxide (TMAO)-producing bacteria. Elevated TMAO is associated with increased mortality and hospitalization in HF, whereas SCFAs enhance barrier integrity and immune tolerance. Secondary bile acids and uremic toxins such as indoxyl sulfate and p-cresyl sulfate further link dysbiosis to fibrosis and vascular stiffness. Circulating markers such as TMAO, lipopolysaccharide-binding protein (LBP), and soluble CD14 carry prognostic value beyond traditional cardiac biomarkers. This review highlights current experimental, translational, and clinical evidence describing gut dysbiosis and its molecular links to HF progression. Targeting the gut–heart axis represents a novel therapeutic approach in HF. Dietary modulation, probiotics/prebiotics, fecal microbiota transplantation, and inhibitors of microbial metabolic pathways show promise. Future research should emphasize microbiota-based interventions in HF management. Full article

(This article belongs to the Special Issue Metabolite Profiles in Inflammatory Diseases)

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18 pages, 3744 KB

Open AccessEditor’s ChoiceArticle

MSTune: A Data-Driven Approach to Parameter Tuning Using Grid Search and Differential Evolution for Gas Chromatography–Mass Spectrometry-Based Compound Identification

by Hunter Dlugas, Jing Li, Xiang Zhang and Seongho Kim

Metabolites 2026, 16(6), 428; https://doi.org/10.3390/metabo16060428 - 18 Jun 2026

Viewed by 402

Abstract

Background/Objectives: In gas chromatography–mass spectrometry (GC-MS) library-based compound identification, spectrum preprocessing and associated tuning parameters critically influence identification performance. These parameters are conventionally optimized using grid search, which requires predefined parameter spaces and becomes computationally inefficient as dimensionality increases, often failing to [...] Read more.

Background/Objectives: In gas chromatography–mass spectrometry (GC-MS) library-based compound identification, spectrum preprocessing and associated tuning parameters critically influence identification performance. These parameters are conventionally optimized using grid search, which requires predefined parameter spaces and becomes computationally inefficient as dimensionality increases, often failing to identify optimal values because of discretization. Differential evolution (DE), a population-based metaheuristic optimization algorithm, provides a flexible alternative through efficient global exploration of the parameter space. This study compared the performance of DE and grid search for optimizing compound identification. Methods: Cosine similarity was applied to the NIST GC-MS library. DE was used to maximize either cross-validated accuracy or mean reciprocal rank (MRR). Results were compared with those from a grid search over five equally spaced parameter values. Identification performance was evaluated using accuracy, MRR, and area under the receiver operating characteristic curve (AUC). Results: When all four parameters were optimized simultaneously, DE achieved slightly higher cross-validated accuracy and MRR than grid search, although the absolute differences were modest. More pronounced differences were observed in specific unidimensional tuning scenarios, particularly for the intensity weight factor. Simultaneous multidimensional parameter optimization yielded better performance than isolated parameter tuning. Conclusions: Grid search may be computationally advantageous when the parameter space is known and limited, whereas DE provides a more flexible approach for unknown or high-dimensional search spaces. Overall, DE achieved comparable identification performance to grid search, with modest improvements observed in some optimization settings. A command line Julia-based tool, MSTune, was developed for spectrum preprocessing parameter optimization and is publicly available on GitHub. Full article

(This article belongs to the Special Issue Open-Source Software in Metabolomics, 2nd Edition)

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16 pages, 590 KB

Open AccessEditor’s ChoiceReview

Ceramide-Driven Mechanisms in Pulmonary Fibrosis

by Zifan Li, Yaqian Li, Na Mao, Xuemin Gao, Hong Xu, Wenchen Cai and Tian Li

Metabolites 2026, 16(6), 421; https://doi.org/10.3390/metabo16060421 - 16 Jun 2026

Cited by 1 | Viewed by 801

Abstract

Pulmonary fibrosis, particularly idiopathic pulmonary fibrosis (IPF), is a chronic and progressive interstitial lung disease characterized by alveolar epithelial injury, fibroblast activation, and excessive extracellular matrix deposition, which collectively lead to respiratory failure. Despite the availability of antifibrotic agents, disease-modifying therapies remain limited. [...] Read more.

Pulmonary fibrosis, particularly idiopathic pulmonary fibrosis (IPF), is a chronic and progressive interstitial lung disease characterized by alveolar epithelial injury, fibroblast activation, and excessive extracellular matrix deposition, which collectively lead to respiratory failure. Despite the availability of antifibrotic agents, disease-modifying therapies remain limited. Emerging evidence has identified dysregulated sphingolipid metabolism, especially ceramide accumulation, as a key driver of fibrotic pathogenesis. Ceramide is a central bioactive lipid in the sphingolipid pathway that regulates multiple cellular processes, including apoptosis, inflammation, endothelial barrier dysfunction, and fibroblast activation, all of which contribute to pulmonary fibrosis. This review is a narrative review that systematically summarizes the biosynthetic and metabolic pathways of ceramide, with an emphasis on chain length-specific functions and the ceramide to S1P rheostat. We further discuss the mechanistic roles of ceramide in alveolar epithelial cell apoptosis, inflammatory responses, and vascular barrier disruption in fibrotic lung disease. Finally, we highlight emerging therapeutic strategies that target ceramide metabolism, including inhibitors of acid sphingomyelinase (ASMase) and serine palmitoyltransferase (SPT), and propose future directions for clinical translation. Full article

(This article belongs to the Special Issue Advances in Immune Metabolism: Lipid Regulation and Disease Outcomes)

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17 pages, 6525 KB

Open AccessEditor’s ChoiceArticle

Melatonin Modulates Macrophage Polarization and Immunometabolic Responses in the Colostrum of Obese Mothers

by Silvia Hannah Bilotti Ratto Gomes da Silva, Danielle Cristina Honorio França, Kênia Maria Resende Silva, Emanuelle Carolina Honorio França, Viviane Francelina Luz, Arce dos Santos Sfredo, Tassiane Cristina Morais, Eduardo Luzía França and Adenilda Cristina Honorio-França

Metabolites 2026, 16(6), 420; https://doi.org/10.3390/metabo16060420 - 15 Jun 2026

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Abstract

Background/Objectives: Obesity is a major public health problem associated with chronic inflammation and functional alterations in multiple organs and systems. Few studies have examined colostrum from obese mothers, particularly with respect to macrophage function, enzyme and cytokine concentrations, and the role of [...] Read more.

Background/Objectives: Obesity is a major public health problem associated with chronic inflammation and functional alterations in multiple organs and systems. Few studies have examined colostrum from obese mothers, particularly with respect to macrophage function, enzyme and cytokine concentrations, and the role of melatonin in immune modulation. This study aimed to evaluate melatonin levels and their effects on macrophage polarization, cytokine concentrations, nitric oxide synthase [iNOS], and arginase in colostrum from obese mothers. Colostrum samples were collected from eutrophic mothers [BMI: 18.5–24.9 kg/m²] and obese mothers [BMI: ≥30 kg/m²]. Methods: Macrophages were isolated by density gradient and treated with melatonin. The expression of M1 and M2 macrophages and cytokine concentrations were assessed by flow cytometry, while melatonin levels in colostrum supernatants, iNOS, and arginase in cell lysates were determined by ELISA. Results: An endogenous increase in melatonin was also observed in the colostrum of obese mothers. Maternal obesity has been shown to reduce M1 and M2 macrophage expression, increase nitric oxide synthase [NOS] activity, and elevate interleukin-6 [IL-6] and interleukin-17 [IL-17] levels. However, melatonin treatment restored M1 and M2 macrophage levels and reduced inducible nitric oxide synthase [iNOS] and arginase production to levels similar to those observed in mothers of healthy weight. Conclusions: these findings suggest that maternal obesity creates a pro-inflammatory environment in colostrum, characterized by altered macrophage polarization, altered cytokine secretion, and an imbalance in the enzymatic activities of iNOS and arginase within the L-arginine metabolic pathway. Both natural and supplemental melatonin exhibited immunomodulatory, antioxidant, and anti-inflammatory effects, helping to restore immune balance in colostrum. These results emphasize the potential benefits of melatonin as an immunometabolic modulator and its contribution to understanding immunometabolic regulation in obese mothers. Full article

(This article belongs to the Special Issue Effects of Nutrition Intake and Lifestyle Intervention on Human Metabolic Health)

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19 pages, 347 KB

Open AccessEditor’s ChoiceReview

Roles of Metabolites Unveiled by Metabolomics in Brassica rapa, B. napus and B. juncea

by Yunong Xia, Silin Su, Xianyu Tang, Lei Qin, Junxing Lu and Shitou Xia

Metabolites 2026, 16(6), 417; https://doi.org/10.3390/metabo16060417 - 15 Jun 2026

Cited by 1 | Viewed by 888

Abstract

Rapeseed is a major source of vegetable oil and contains a wide variety of metabolites. Recent advances, particularly the integration of metabolomics with other omics approaches, have enabled not only comprehensive but also detailed analyses of key metabolites that respond to specific conditions. [...] Read more.

Rapeseed is a major source of vegetable oil and contains a wide variety of metabolites. Recent advances, particularly the integration of metabolomics with other omics approaches, have enabled not only comprehensive but also detailed analyses of key metabolites that respond to specific conditions. To date, these recent advances in the metabolomics of Brassica crops have not yet been fully clarified. In this review, we seek to summarize the recent progresses in metabolomics studies of Brassica rapa, B. napus and B. juncea, introduce the key metabolites spanning nucleic acids, amino acids, fatty acids, lipids, organic acids, alkaloids, phenylpropanoids, terpenoids, flavonoids and glucosinolates uncovered by this approach, focusing on those associated with growth and development, and abiotic/biotic stresses, including macronutrient availability, temperature, water stress, salt stress, aluminum and cadmium toxicity, and infection of Sclerotinia sclerotiorum, Leptosphaeria maculans, and Plasmodiophora brassicae. Future perspectives and current challenges in metabolomics integrating with other omics are also discussed, along with its potential for breeding applications, especially in new marker discovery, trait prediction, and even metabolic selection, aimed at developing new rapeseed varieties with stable, high-yielding, and quality traits. Full article

(This article belongs to the Special Issue Metabolomics and Plant Defence, 2nd Edition)

27 pages, 3122 KB

Open AccessEditor’s ChoiceReview

Exploring the Health Effects of Phytoestrogens

by Vladimír Kraus, Anna Birková, Miroslava Majerníková and Beáta Čižmárová

Metabolites 2026, 16(6), 410; https://doi.org/10.3390/metabo16060410 - 12 Jun 2026

Viewed by 669

Abstract

Background/Objectives: Phytoestrogens are secondary plant metabolites produced via the phenylpropanoid pathway. They include a broad spectrum of chemical compounds, such as phenolics, flavonoids, isoflavones, coumestans, lignans, and others. Their chemical structures resemble those of estradiol, and they exhibit biological effects similar to those [...] Read more.

Background/Objectives: Phytoestrogens are secondary plant metabolites produced via the phenylpropanoid pathway. They include a broad spectrum of chemical compounds, such as phenolics, flavonoids, isoflavones, coumestans, lignans, and others. Their chemical structures resemble those of estradiol, and they exhibit biological effects similar to those of human estrogens, influencing many physiological processes throughout life in both men and women—including the timing and progression of puberty. Methods: The literature search included databases such as PubMed, Scopus, Web of Science, and Google Scholar with the use of specific keywords. Studies were considered eligible if they reported original findings from observational studies (cohort, case–control, and cross-sectional) or from experimental studies. Results: Phytoestrogens can modulate estrogenic activity and interact with a variety of biological pathways. These compounds may play a role in human development and pubertal processes, contribute to overall health, and potentially help alleviate menopausal symptoms and reduce the risk of certain cancers. Conclusions: Phytoestrogens have numerous positive effects on the human body across various stages of life. Their overall impact and potency, however, seem to be influenced by factors such as intake level, individual genetic variability, and the specific phytoestrogen class consumed. Full article

(This article belongs to the Special Issue Effects of Secondary Plant Metabolites on Human Health)

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24 pages, 2738 KB

Open AccessEditor’s ChoiceReview

Phytohormonal Regulation of Plant Responses to Major Abiotic Stresses: From Signaling Pathways to Hormonal Crosstalk

by Shadi Sadat Mehrabi, Manijeh Sabokdast and Beata Dedicova

Metabolites 2026, 16(6), 401; https://doi.org/10.3390/metabo16060401 - 9 Jun 2026

Cited by 1 | Viewed by 798

Abstract

Plants are constantly exposed to diverse abiotic stresses, including drought, salinity, and extreme temperatures, which severely limit growth, development, and crop productivity. These stresses disrupt physiological, biochemical, and molecular processes, leading to reduced photosynthesis, altered water and ion homeostasis, and accumulation of reactive [...] Read more.

Plants are constantly exposed to diverse abiotic stresses, including drought, salinity, and extreme temperatures, which severely limit growth, development, and crop productivity. These stresses disrupt physiological, biochemical, and molecular processes, leading to reduced photosynthesis, altered water and ion homeostasis, and accumulation of reactive oxygen species (ROS). Plants have evolved sophisticated sensing and signaling mechanisms to perceive these stresses, with phytohormones playing central roles in mediating adaptive responses. Key hormones, including abscisic acid (ABA), salicylic acid (SA), jasmonates (JAs), gibberellins (GAs), auxin (IAA), ethylene (ET), melatonin, and strigolactones (SLs), regulate stress tolerance by controlling stomatal behavior, root architecture, antioxidant systems, osmolyte accumulation, and stress-responsive gene expression. Importantly, these hormones operate within an intricate network of crosstalk, integrating multiple signaling pathways to balance growth and stress adaptation. Interactions among ABA, GA, JA, SA, auxin, ET, SLs, and melatonin enable plants to coordinate transcriptional regulation, protein phosphorylation, and ROS signaling, optimizing survival under fluctuating environmental conditions. Understanding the molecular mechanisms underlying hormonal crosstalk and their roles in abiotic stress tolerance provides valuable insights for developing resilient crops in the face of climate change. Full article

(This article belongs to the Special Issue Climate Change-Related Stresses and Plant Metabolism)

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26 pages, 8195 KB

Open AccessEditor’s ChoiceReview

A Chrono-Metabolic Approach to Mental Health: Current Perspectives on Circadian Rhythms, Gut Microbiota, and Microbial Metabolites in Mood Disorders

by Giuseppe Marano, Mariateresa Acanfora, Luca Conci, Gianandrea Traversi, Osvaldo Mazza, Esmeralda Capristo, Eleonora Gaetani, Gianluca Franceschini and Marianna Mazza

Metabolites 2026, 16(6), 400; https://doi.org/10.3390/metabo16060400 - 9 Jun 2026

Viewed by 969

Abstract

Growing evidence indicates that the gut microbiota is not a static ecosystem but a rhythmic metabolic organ whose oscillatory activity is tightly coordinated with host circadian biology. Disruption of this temporal alignment, through irregular diet, sleep disturbance, shift work, or social jet lag, [...] Read more.

Growing evidence indicates that the gut microbiota is not a static ecosystem but a rhythmic metabolic organ whose oscillatory activity is tightly coordinated with host circadian biology. Disruption of this temporal alignment, through irregular diet, sleep disturbance, shift work, or social jet lag, may profoundly alter microbial composition and the production of neuroactive metabolites. These alterations have emerged as potential contributors to the pathophysiology of mood disorders. This review introduces the concept of chrono-metabolic psychiatry, a framework integrating circadian rhythms, gut microbiota dynamics, and host metabolic signaling in the development and course of depressive and bipolar disorders. In this framework, the term “chrono-metabolic” refers to the integration of biological timing, host metabolic regulation, and microbiota-derived metabolic signaling. Chrono-metabolic psychiatry therefore shifts the focus from static dysbiosis or neurotransmitter imbalance alone to the time-dependent interactions among circadian misalignment, microbial rhythmicity, immune regulation, metabolite production, and affective instability. Diurnal fluctuations in short-chain fatty acids, tryptophan–kynurenine metabolites, bile acids, and microbial-derived neurotransmitters interact with clock gene regulation, hypothalamic–pituitary–adrenal axis activity, neuroinflammation, and synaptic plasticity. Chrono-disruption may represent a transdiagnostic vulnerability factor and may confirm the bidirectional relationship between mood instability and microbiota rhythmicity. Emerging therapeutic implications, including chrono-nutrition, time-restricted feeding, targeted probiotic administration (“chronobiotics”), and the microbiota-modulating effects of psychotropic medications are discussed. By shifting from a compositional to a temporal–metabolic perspective, this model highlights the importance of microbial oscillations rather than static dysbiosis alone. Integrating circadian biology into microbiota research may enable metabolomic stratification and pave the way for precision psychiatry approaches grounded in host–microbe metabolic crosstalk. Future longitudinal and time-resolved multi-omics studies are needed to validate this framework and to translate it into clinically actionable interventions. Full article

(This article belongs to the Special Issue Modulating Gut Microbiota and Host Interactions: The Role of Diet, Probiotics, and Circadian Rhythms)

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16 pages, 2623 KB

Open AccessEditor’s ChoiceArticle

Rapid LC-MS/MS Method for Targeted Assay of Creatine Deficiency Syndromes in Morocco

by Faïza Meiouet and François Boemer

Metabolites 2026, 16(6), 388; https://doi.org/10.3390/metabo16060388 - 3 Jun 2026

Viewed by 426

Abstract

Background: Creatine deficiency syndromes (CDS) are rare neurometabolic disorders caused by defects in creatine biosynthesis (AGAT and GAMT deficiencies) or creatine transport (SLC6A8 deficiency). Early biochemical recognition is crucial for timely treatment of AGAT and GAMT deficiencies and for improving neurodevelopmental outcomes. In [...] Read more.

Background: Creatine deficiency syndromes (CDS) are rare neurometabolic disorders caused by defects in creatine biosynthesis (AGAT and GAMT deficiencies) or creatine transport (SLC6A8 deficiency). Early biochemical recognition is crucial for timely treatment of AGAT and GAMT deficiencies and for improving neurodevelopmental outcomes. In Morocco, expanding the liquid chromatography-tandem mass spectrometry (LC-MS/MS) biomarker panel for inherited metabolic disorders is a priority to strengthen diagnostic capacity and reduce diagnostic delay. Methods: We developed and validated a rapid LC-MS/MS method for the simultaneous quantification of creatine (Cr), guanidinoacetate (GAA), and creatinine (Crn) in plasma and urine using isotopically labelled internal standards and a standardized sample preparation procedure. Analytical performance, including linearity, precision, accuracy, sensitivity, matrix effects, carryover, inter-sample contamination, stability, and measurement uncertainty, was assessed in accordance with ISO 15189:2022 requirements. Results: The assay showed excellent linearity across the analytical range (r² > 0.99), with robust intra- and inter-day precision (CV < 10%). Limits of detection (LOD) were 0.05 µmol/L for Cr and 0.03 µmol/L for GAA in urine, and 0.05 µmol/L for Cr and GAA in plasma. The total run time was 1.1 min per sample, supporting high-throughput implementation. Method performance was further supported by satisfactory results in ERNDIM external quality assessment schemes. Preliminary internal reference ranges and expanded measurement uncertainty were calculated from the available anonymized dataset. Conclusions: This rapid LC-MS/MS method enables the measurement of key CDS biomarkers and contributes to expanding the LC-MS/MS biomarker panel for inherited metabolic disorders in Morocco. Full article

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

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19 pages, 3861 KB

Open AccessEditor’s ChoiceReview

Circadian Regulation of Gut Microbial Metabolites in Intestinal Epithelial Homeostasis

by Miri Park, Sooah Kim and Jeehwan Choe

Metabolites 2026, 16(6), 383; https://doi.org/10.3390/metabo16060383 - 1 Jun 2026

Viewed by 578

Abstract

The gut microbiota produces chemically diverse metabolites whose levels fluctuate depending on the time of day, driven by bidirectional coupling between host intestinal circadian clocks and intrinsic microbial oscillators. Although short-chain fatty acids have received the most attention as microbial circadian effectors, a [...] Read more.

The gut microbiota produces chemically diverse metabolites whose levels fluctuate depending on the time of day, driven by bidirectional coupling between host intestinal circadian clocks and intrinsic microbial oscillators. Although short-chain fatty acids have received the most attention as microbial circadian effectors, a broad class of metabolites, including secondary bile acids, indole derivatives, and branched-chain fatty acids, engage distinct epithelial receptors and transcriptional programs through mechanisms that are, to varying degrees, subject to circadian regulation. However, the mechanisms by which these metabolite classes collectively regulate barrier integrity, mucosal immune tone, and stem cell-driven renewal, as well as the consequences of their rhythmicity loss under circadian misalignment, have not been systematically reviewed. This review constructs a mechanistic framework linking microbial metabolite rhythmicity to the circadian regulation of intestinal epithelial homeostasis and evaluates dietary and probiotic interventions that modulate this axis as chronobiotic strategies. Convergent mechanisms, unresolved questions, and translational opportunities are identified across in vitro, preclinical, and clinical evidence. Full article

(This article belongs to the Special Issue Modulating Gut Microbiota and Host Interactions: The Role of Diet, Probiotics, and Circadian Rhythms)

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11 pages, 1034 KB

Open AccessEditor’s ChoiceArticle

Humanin Restores Metabolic Hormone Homeostasis of Leptin, Ghrelin, Irisin and Asprosin in Streptozotocin-Induced Diabetic Mice

by Ferah Bulut, Muhammed Adam, Aslısah Ozgen and Mete Ozcan

Metabolites 2026, 16(6), 373; https://doi.org/10.3390/metabo16060373 - 29 May 2026

Cited by 1 | Viewed by 634

Abstract

Objective: Diabetes mellitus is closely associated with mitochondrial dysfunction, which disrupts cellular energy metabolism and perturbs hormonal homeostasis. Humanin (HN), a 24-amino acid peptide encoded within the mitochondrial genome, has attracted considerable attention due to its cytoprotective and metabolic regulatory properties. Despite [...] Read more.

Objective: Diabetes mellitus is closely associated with mitochondrial dysfunction, which disrupts cellular energy metabolism and perturbs hormonal homeostasis. Humanin (HN), a 24-amino acid peptide encoded within the mitochondrial genome, has attracted considerable attention due to its cytoprotective and metabolic regulatory properties. Despite its recognized biological potential, the role of HN in coordinating key metabolic hormone networks under diabetic conditions remains poorly understood. This study aimed to investigate the integrative effects of repeated humanin administration on key metabolic hormones leptin, ghrelin, irisin, and asprosin and its potential role in restoring hormonal homeostasis in a streptozotocin (STZ)-induced diabetic mouse model. Materials and Methods: Forty male mice were randomly assigned to four groups (n = 10 for each group): control, HN (4 mg/kg/day), STZ (150 mg/kg), and STZ + HN. Humanin was administered intraperitoneally for 15 consecutive days. Serum levels of leptin, asprosin, irisin, and ghrelin were measured using enzyme-linked immunosorbent assay (ELISA), and data were analyzed using one-way ANOVA followed by Tukey’s post hoc test. Results: STZ-induced diabetes markedly disrupted metabolic hormone balance, as indicated by decreased leptin and irisin levels and increased asprosin concentrations. Repeated HN treatment effectively restored leptin levels and suppressed asprosin concentrations, while irisin levels showed a relative increase compared to the STZ animals. In addition, ghrelin levels were significantly elevated in HN-treated diabetic mice compared to untreated STZ animals. Conclusions: These findings indicate that humanin exerts an integrative, multi-hormonal regulatory effect, supporting the restoration of metabolic and endocrine homeostasis under diabetic conditions. Full article

(This article belongs to the Section Pharmacology and Drug Metabolism)

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18 pages, 27428 KB

Open AccessEditor’s ChoiceArticle

Integrating Sensory Evaluation and Metabolomics to Reveal the Metabolic Basis of Taste and Flesh Color in Melon (Cucumis melo L.)

by Yu Zhou, Binbin Li, Weizhong He, Fengjuan Liu, Yingying Fan, Jiangtao Du, Xing Cui, Weijia Lian, Qi Shen, Yan Wang, Zhongkai Zhao and Cheng Wang

Metabolites 2026, 16(6), 368; https://doi.org/10.3390/metabo16060368 - 28 May 2026

Viewed by 522

Abstract

Background: The sensory quality of melon (Cucumis melo L.) is determined by the complex interplay of metabolites within the fruit. However, the underlying metabolic mechanisms based on consumer sensory experience remain underexplored. Methods: Sensory evaluation was conducted on twelve melon [...] Read more.

Background: The sensory quality of melon (Cucumis melo L.) is determined by the complex interplay of metabolites within the fruit. However, the underlying metabolic mechanisms based on consumer sensory experience remain underexplored. Methods: Sensory evaluation was conducted on twelve melon cultivars, recording flesh color and quantitatively scoring acidity, sweetness, firmness, and aroma intensity. Based on the sensory results, eight cultivars were selected to establish two contrasting groups: sweet-type vs. acidic-type and orange-fleshed vs. green-fleshed. Untargeted metabolomics (UPLC-QTOF-MS) was then performed to analyze the samples, and differential metabolites were screened using OPLS-DA combined with univariate analysis. Results: Pathway enrichment analysis revealed that the key distinction between sweet and acidic taste profiles was associated with the specific accumulation of citric acid within the tricarboxylic acid (TCA) cycle in the acidic-type group. Regarding flesh color, the orange-fleshed group was enriched with carotenoid derivatives like β-citraurinene and the oxidized tocopherol product α-tocopherolquinone, whereas the green-fleshed group mainly accumulated phytol, a chlorophyll degradation product, along with more abundant terpenoids. Conclusions: By integrating sensory phenotyping with metabolomic analysis, this study identified key differential metabolites and candidate pathways associated with taste and color in melon, providing metabolic insights and data resources for quality evaluation and regulation. Full article

(This article belongs to the Section Food Metabolomics)

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19 pages, 4815 KB

Open AccessEditor’s ChoiceArticle

Metabolic Responses of Grapevine Leaves to Grapevine Leafroll-Associated Virus 3 Infection

by Ivana Tomaz, Darko Preiner, Nina Buljević and Darko Vončina

Metabolites 2026, 16(6), 359; https://doi.org/10.3390/metabo16060359 - 27 May 2026

Viewed by 609

Abstract

Background: Grapevine leafroll-associated virus 3 (GLRaV-3) is a severe phloem-limited grapevine virus, meaning it is restricted to sugar-transporting vascular tissue, which helps explain its strong effects on leaf physiology and carbon transport. However, its impact on leaf oxidative status, phenolic composition, and [...] Read more.

Background: Grapevine leafroll-associated virus 3 (GLRaV-3) is a severe phloem-limited grapevine virus, meaning it is restricted to sugar-transporting vascular tissue, which helps explain its strong effects on leaf physiology and carbon transport. However, its impact on leaf oxidative status, phenolic composition, and volatile organic compounds (VOCs) remains insufficiently characterized. Methods: Virus-free and GLRaV-3-infected grapevine leaves were analyzed for photosynthetic pigments, oxidative stress markers, phenolic compounds, and VOCs using spectrophotometric assays, HPLC-DAD/FLD, and SPME-Arrow-GC/MS. Data were evaluated using one-way ANOVA, multiple testing correction, principal component analysis (PCA), and exploratory partial least squares-discriminant analysis (PLS-DA). Results: GLRaV-3-infected leaves showed lower chlorophyll a (576.75 vs. 657.85 mg 100 g⁻¹ DW), chlorophyll b (282.96 vs. 314.05 mg 100 g⁻¹ DW), and total carotenoids (125.89 vs. 154.65 mg 100 g⁻¹ DW), but higher malondialdehyde (11.91 vs. 8.73 nmol g⁻¹ DW), H₂O₂ (0.36 vs. 0.25 μmol g⁻¹ DW), and proline (8.83 vs. 7.98 μmol g⁻¹ DW). Phenolic profiling showed increased levels of several flavonols and hydroxycinnamic acids, including kaempferol-3-O-glucuronide (2.81-fold), myricetin-3-O-glucoside (1.75-fold), quercetin-3-O-glucuronide (1.48-fold), and caffeic acid (1.30-fold). VOC profiling revealed higher relative abundances of several green leaf volatile-related compounds and methyl salicylate, including 1-methoxy-2-propanol (1.85-fold), 1-penten-3-ol (1.58-fold), hexanal (1.42-fold), and methyl salicylate (1.37-fold). PCA summarized treatment-related differences, with the first two components explaining 63.73% of phenolic and 74.09% of VOC variability, while exploratory PLS-DA/VIP analysis further supported the identification of treatment-associated discriminant metabolic features. Conclusions: GLRaV-3 infection is associated with reduced pigment content, increased oxidative stress markers, and coordinated changes in phenolic and VOC profiles. These metabolite changes provide insight into grapevine responses to viral infection and highlight GLRaV-3-associated metabolic features for future targeted studies of grapevine leafroll disease. Full article

(This article belongs to the Special Issue Plant Biotic and Abiotic Stress Responses and Tolerance: Phytohormonal and Metabolic Insights)

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19 pages, 447 KB

Open AccessEditor’s ChoiceArticle

Chemical Structure Representation Standardization Is Needed to Generalize Metabolite-Pathway Involvement Prediction Across KEGG, Reactome, and MetaCyc Knowledgebases

by Erik D. Huckvale and Hunter N. B. Moseley

Metabolites 2026, 16(6), 357; https://doi.org/10.3390/metabo16060357 - 26 May 2026

Cited by 1 | Viewed by 662

Abstract

Background/Objectives: Due to the utility of knowing the pathway involvement of metabolites detected in biological experiments, knowledgebases such as the Kyoto Encyclopedia of Genes and Genomes (KEGG), Reactome, and MetaCyc have annotated compound entries to specific pathways defined by the knowledgebase. However, [...] Read more.

Background/Objectives: Due to the utility of knowing the pathway involvement of metabolites detected in biological experiments, knowledgebases such as the Kyoto Encyclopedia of Genes and Genomes (KEGG), Reactome, and MetaCyc have annotated compound entries to specific pathways defined by the knowledgebase. However, these compound-pathway annotations are largely incomplete and are costly to obtain experimentally or curate from published scientific literature. This metabolite-pathway annotation incompleteness problem is amenable to machine learning (ML)-based solutions. But to date, no machine learning model has been trained on all three knowledgebases to maximize its performance and robustness. This may be due to inconsistencies in chemical structure representation that can confuse a model and greatly reduce generalizability. Methods: We constructed a new training dataset with roughly 50,000,000 entries using compound-pathway annotations derived from KEGG, Reactome, and MetaCyc. We trained and tested a multitask classification, graph convolutional neural network-like model that classifies compound involvement with 8056 pathways that have unique pathway representations, based on annotated compound chemical structures represented with chemical substructure features. While the initial dataset contained inconsistencies in chemical structure representations across knowledgebases, we alleviated this issue by standardizing chemical structure representation using InChI (IUPAC International Chemical Identifier) canonicalization. We compared the performance of the non-standardized versus the standardized dataset and quantified their generalizability by comparing training set compounds to their knowledgebase cross-references. Results: While the non-standardized dataset scored a mean Matthews correlation coefficient (MCC) of 0.8725 ± 0.0064, the standardized dataset scored an MCC of 0.9036 ± 0.0033. When comparing model generalizability, the non-standardized chemical structure representations had a huge 0.2687 drop in mean MCC, while the standardized chemical structure representations only had a 0.0384 drop in mean MCC. Conclusions: We constructed the largest ML-ready dataset for predicting compound-pathway involvement to date. Next, we constructed, trained, and evaluated the highest performing ML model capable of predicting the highest number of pathway annotations to date. We discovered that standardizing chemical structure representation is an essential step when predicting novel chemical structures. Full article

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

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17 pages, 961 KB

Open AccessEditor’s ChoiceArticle

Detection of Nitazenes (2-Benzylbenzimidazoles)—Novel Synthetic Opioids in Coronial Casework in Victoria, Australia

by Lachlan Scully, Jared W. Castle, Matthew Di Rago, Hans H. de Boer, Jennifer Schumann, Kerryn Crump, Linda Glowacki and Dimitri Gerostamoulos

Metabolites 2026, 16(6), 358; https://doi.org/10.3390/metabo16060358 - 26 May 2026

Viewed by 942

Abstract

Background: The 2-benzylbenzimidazoles or nitazenes are an evolving class of highly potent mu-opioid receptor agonists. Nitazenes were originally developed in the late 1950s for pharmaceutical use as analgesics; however, due to their extreme potency and the risk of adverse health outcomes, pharmaceutical [...] Read more.

Background: The 2-benzylbenzimidazoles or nitazenes are an evolving class of highly potent mu-opioid receptor agonists. Nitazenes were originally developed in the late 1950s for pharmaceutical use as analgesics; however, due to their extreme potency and the risk of adverse health outcomes, pharmaceutical research was discontinued. Since 2019, nitazenes have emerged as illicit drugs of abuse, causing significant concern. From 2021, they have been detected in both coronial and clinical casework in Victoria, Australia. This study examined nitazene-related coronial casework in Victoria from 2021 to 2025 to explore the trends and characteristics of nitazene-related deaths. Methods: Relevant cases were identified from the Victorian Institute of Forensic Medicine’s (VIFM’s) case management system. Data were collated and analysed from all coronial cases where a nitazene was detected by a toxicological analysis between 1 January 2021 and 31 December 2025. Trend comparisons were made with nitazene detections reported in other countries. Results: Nitazenes were detected in 23 deaths from a total of approximately 33,108 coronial cases admitted to the VIFM for investigation over the time period. The age range was 17–45 years, with a median of 32 and with 87% of the deaths being male. The nitazenes detected were protonitazene (n = 14), metonitazene (n = 5), isotonitazene (n = 2), N-pyrrolidino etonitazene (n = 2), N-desethyl isotonitazene (n = 1), methylenedioxynitazene (n = 1) and etodesnitazene (n = 1). Two cases contained more than one nitazene; both involved protonitazene, one involved metonitazene, and the other involved N-desethyl isotonitazene and methylenedioxynitazene. The timeline of detection of these nitazenes displays similarities with emergence trends in other countries. The nitazene concentrations ranged from 0.1 to 33 ng/mL. Broad polydrug usage was evident in all cases, with other drugs co-detected in the blood including stimulants (particularly, methylamphetamine (48%) and cocaine (44%)) as well as pharmaceutical benzodiazepines (43%) and pharmaceutical opioids (22%), and 13% had 6-monoacetylmorphine detected in either blood or urine. Novel benzodiazepines (39%) were also common, including bromazolam, which was co-detected in 35% of cases. Nineteen deaths were attributed solely to nitazene-related mixed-drug toxicity, while the remaining four cases were attributed to cardiac- and pulmonary-related disease, with polydrug use deemed a contributing factor. Conclusions: This novel case series adds comprehensive toxicological information to the body of evidence reinforcing the high risk of harm associated with the use of nitazenes. It is imperative that toxicology services continue to monitor for nitazenes to promote community awareness against nitazene-related harm. Full article

(This article belongs to the Special Issue Toxicodynamics and Toxicokinetics of New Psychoactive Substances: From In Vitro Insights to In Vivo Impact)

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15 pages, 3182 KB

Open AccessEditor’s ChoiceArticle

Effects of Long-Term Administration of Ginsenosides on the Structure of Intestinal Microflora and the Absorption and Utilization of Saponins in Rats

by Guoliang Dou, Xinyue Bi, Nuoyan Wang, Shuhang Li, Yu Huang, Shirui Lu, Zixin Wang, Wenxiu Ji, Ye Hong and Weiwei Dong

Metabolites 2026, 16(6), 352; https://doi.org/10.3390/metabo16060352 - 25 May 2026

Viewed by 461

Abstract

Background: Ginsenosides are active natural compounds with diverse effects, and their interaction with the gut microbiota can influence microbial composition and abundance, though the long-term effects remain unclear. Methods: This study examines the impact of long-term oral ginsenoside administration on gut microbiota composition [...] Read more.

Background: Ginsenosides are active natural compounds with diverse effects, and their interaction with the gut microbiota can influence microbial composition and abundance, though the long-term effects remain unclear. Methods: This study examines the impact of long-term oral ginsenoside administration on gut microbiota composition and structure in rats, as well as its pharmacokinetics. Twenty healthy male Wistar rats were divided into a control group (CK, receiving distilled water) and a ginsenoside treatment group (PGE, 100 mg/kg) for 30 days. Fecal samples were analyzed using 16S rRNA high-throughput sequencing on the Illumina HiSeq platform to assess microbial diversity. Concurrently, liquid chromatography–tandem mass spectrometry (LC-MS/MS) was utilized to determine the concentrations of ginsenosides in the serum and to investigate their pharmacokinetic properties (p < 0.05). Results: The results indicated that the α-diversity indices of the gut microbiota in the PGE group were significantly higher than in the CK group, suggesting that ginsenosides enhance microbial richness and diversity (p < 0.05). At the phylum level, the relative abundance of Firmicutes in the PGE group increased by 10.6% ± 2.72%, while that of Bacteroidetes decreased by 11.5% ± 3.18%; at the genus level, the proportion of Lactobacillus genus rose by 17.78% ± 4.37% (p < 0.05). Pharmacokinetic analysis revealed that the area under the concentration–time curve (AUC) and maximum concentration (C_max) of ginsenosides were significantly higher in the PGE group than in the CK group. Conclusions: Chronic oral administration of ginsenosides improves their absorption and utilization through gut microbiota modulation, offering experimental evidence for deeper insight into ginsenoside–microbe interactions. Full article

(This article belongs to the Special Issue Drug Metabolism, Interactions and Toxicity: Advances in Metabolomics Approaches)

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16 pages, 11209 KB

Open AccessEditor’s ChoiceArticle

Geographic Variation and Quality Consistency of Toddaliae Asiaticae Radix: A Hybrid Framework Integrating Environmental Feature and Bioactivity-Weighted Modeling

by Linjiang Wei, Hong Chen, Mengmeng Sun, Yuanle Song, Chen Zhang and Zhi Zhou

Metabolites 2026, 16(6), 353; https://doi.org/10.3390/metabo16060353 - 25 May 2026

Viewed by 379

Abstract

Background: Toddaliae Asiaticae Radix (TA) boasts a long history of medicinal application. However, origin traceability and quality assessment of the widely distributed original plant Toddalia asiatica have not been fully elucidated. Methods: A hybrid framework integrating targeted metabolomics, network pharmacology (NP), [...] Read more.

Background: Toddaliae Asiaticae Radix (TA) boasts a long history of medicinal application. However, origin traceability and quality assessment of the widely distributed original plant Toddalia asiatica have not been fully elucidated. Methods: A hybrid framework integrating targeted metabolomics, network pharmacology (NP), and machine learning (ML) was established. By optimizing key parameters, a high-coverage and rapid method for multiple categories compounds was developed using ultra-high performance liquid chromatography-multiple reaction monitoring tandem mass spectrometry (UPLC-MRM MS/MS). Using samples collected across 16 geographical regions, redundancy analysis (RDA) and pattern recognition techniques were applied to explore environment-sensitive metabolites. Taking into account five types of diseases, NP analysis was employed to obtain the bioactive components and their contribution weight in disease treatment. Subsequently, core Quality Markers (Q-Markers) with dual functions of responsive to geographic variations and biologically relevant to therapeutic efficacy were figured out, and were used to establish origin scoring model and discrimination model. Results: The geographical metabolic characteristics of the TA from broad regions in China were thoroughly analyzed, and 60 geographically sensitive compounds were identified. Through NP analysis, 27 core Q-Markers were locked. The bioactivity-weighted scoring model based on Q-Markers revealed the consistency of regional rankings as well as minor fluctuations across five diseases. ML demonstrated that the Q-Markers preserved regional discrimination performance, and the introducing of practical-oriented weights enhanced overall discriminative confidence. Conclusions: This research decodes the Geographical metabolic characteristics of TA, and highlights the necessity of function-oriented prioritization of drug resources. Full article

(This article belongs to the Section Plant Metabolism)

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19 pages, 9086 KB

Open AccessEditor’s ChoiceArticle

Mapping Spatiotemporal Metabolic Perturbations in Alloxan-Induced Diabetic Rat Kidneys Using Spatial Metabolomics and Proteomic Integration

by Tianfang Lan, Caiying Liu, Xingyu Zhang, Xiaoyu Zhang, Yuchen Liu, Wenxuan Shao and Zhonghua Wang

Metabolites 2026, 16(6), 355; https://doi.org/10.3390/metabo16060355 - 25 May 2026

Viewed by 460

Abstract

Background: Diabetic nephropathy (DN) is characterized by complex and region-specific metabolic dysregulation that is not captured by conventional biomarkers. However, the spatiotemporal organization of metabolic alterations across renal compartments in type 1 diabetes remains poorly understood. Methods: In this study, spatial metabolomics based [...] Read more.

Background: Diabetic nephropathy (DN) is characterized by complex and region-specific metabolic dysregulation that is not captured by conventional biomarkers. However, the spatiotemporal organization of metabolic alterations across renal compartments in type 1 diabetes remains poorly understood. Methods: In this study, spatial metabolomics based on air flow-assisted desorption electrospray ionization mass spectrometry imaging (AFADESI-MSI) was applied to investigate metabolic alterations in kidney tissues from alloxan-induced diabetic rats at 4 and 8 weeks post-induction. Complementary LC–MS/MS metabolite profiling and label-free proteomic analysis were performed to support pathway interpretation. Results: Spatial metabolomics revealed pronounced region- and time-dependent metabolic reprogramming in diabetic kidneys. Early-stage (DN-4w) changes were characterized by elevated glucose and activation of glucose-associated pathways, including the polyol pathway, accompanied by accumulation of acylcarnitines and lipid intermediates, indicating metabolic substrate overload. At later stages (DN-8w), glucose and related metabolites declined, reflecting impaired metabolic capacity and mitochondrial dysfunction. Broad remodeling of lipid metabolism, including glycerophospholipids, fatty acids, and hexosylceramide, was observed, along with dysregulation of amino acid metabolism and redox-related pathways. These alterations exhibited clear regional heterogeneity across renal cortex and medulla, highlighting compartment-specific metabolic vulnerability. Conclusions: This study provides a comprehensive spatial characterization of metabolic perturbations during DN progression, revealing coordinated alterations in glucose utilization, lipid metabolism, and mitochondrial function. The findings demonstrate the value of spatial metabolomics in uncovering region-specific metabolic mechanisms and provide new insights into the pathogenesis of diabetic nephropathy. Full article

(This article belongs to the Special Issue Mass Spectrometry Imaging and Spatial Metabolomics—2nd Edition)

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23 pages, 911 KB

Open AccessEditor’s ChoiceReview

Obesity, Low-Grade Chronic Inflammation, and Clinical Outcomes in Spondyloarthritis: A Translational Synthesis

by Andrej Belančić, Mislav Radić, Marija Rogoznica Pavlović, Marijana Vučković, Petra Šimac Prižmić, Elvira Meni Maria Gkrinia, Josipa Radić and Almir Fajkić

Metabolites 2026, 16(5), 347; https://doi.org/10.3390/metabo16050347 - 21 May 2026

Cited by 1 | Viewed by 969

Abstract

This translational synthesis highlights the potential role of obesity-induced low-grade chronic inflammation in modulating clinical outcomes among patients with spondyloarthritis (SpA). Obesity transforms adipose tissue into a pro-inflammatory endocrine organ, where hypertrophic adipocytes release adipokines such as leptin alongside cytokines including TNF-α and [...] Read more.

This translational synthesis highlights the potential role of obesity-induced low-grade chronic inflammation in modulating clinical outcomes among patients with spondyloarthritis (SpA). Obesity transforms adipose tissue into a pro-inflammatory endocrine organ, where hypertrophic adipocytes release adipokines such as leptin alongside cytokines including TNF-α and IL-6, potentially contributing to macrophage polarization toward an M1 phenotype and activating NF-κB signaling pathways. This systemic immunometabolic priming may lower activation thresholds at the enthesis—the primary pathological site in SpA—potentially amplifying IL-23/IL-17 axis activity via Th17 bias, innate-like lymphocyte responses, and stromal–immune crosstalk under mechanical stress. Clinically, patients with SpA and obesity have been reported to demonstrate heightened disease activity (BASDAI, ASDAS), impaired function (BASFI), accelerated radiographic progression (syndesmophytes, enthesophytes), and diminished biologic response rates, potentially attributable to pharmacokinetic alterations (e.g., subtherapeutic TNF inhibitor levels) and pharmacodynamic resistance. Multisystem comorbidities, including non-alcoholic fatty liver disease, cardiovascular events, metabolic syndrome, sleep disturbances, and depression, further exacerbate morbidity and diminish quality of life. Therapeutic implications emphasize obesity as a modifiable disease modifier. Weight loss interventions, including hypocaloric diets, anti-inflammatory regimens (e.g., Mediterranean diet), multicomponent exercise, GLP-1 receptor agonists, and bariatric surgery, have been associated with reductions in inflammatory biomarkers, improved remission rates (MDA, DAPSA), and prolonged drug survival by restoring adipokine balance and disrupting mechano-inflammatory loops. Future randomized controlled trials should prioritize long-term evaluations of integrated multidisciplinary strategies that combine metabolic optimization with immunomodulatory therapies, addressing adherence challenges through psychological support and patient-tailored protocols, while elucidating dose–response relationships for GLP-1RAs and exercise in diverse SpA subtypes to establish precision management paradigms that mitigate cardiometabolic burden and improve holistic outcomes. Full article

(This article belongs to the Section Cell Metabolism)

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25 pages, 1293 KB

Open AccessEditor’s ChoiceReview

Targeting NAD Homeostasis: Compartmentalization, Quantification, and Modulation

by Marta Nobile, Veronica Fontanini, Simone Serrao, Johannes Burtscher, Francesca Re and Giuseppe Paglia

Metabolites 2026, 16(5), 338; https://doi.org/10.3390/metabo16050338 - 18 May 2026

Viewed by 1438

Abstract

Nicotinamide adenine dinucleotide (NAD⁺) and its reduced form, NADH, are essential coenzymes that play central roles in cellular redox homeostasis, energy metabolism, DNA repair, and signaling. Cellular NAD⁺ levels are maintained by a dynamic balance between the de novo Preiss–Handler, [...] Read more.

Nicotinamide adenine dinucleotide (NAD⁺) and its reduced form, NADH, are essential coenzymes that play central roles in cellular redox homeostasis, energy metabolism, DNA repair, and signaling. Cellular NAD⁺ levels are maintained by a dynamic balance between the de novo Preiss–Handler, and salvage synthesis pathways, and consumption by enzymes like sirtuins, PARPs, and CD38. Among these, the nicotinamide Phosphoribosyltransferase (NAMPT)-driven salvage pathway represents the predominant route of NAD+ synthesis. The specific regulation of NAD (NAD⁺ and NADH) levels across distinct subcellular compartments has emerged as a critical determinant of cellular function but it remains poorly understood. Dysregulation of NAD metabolism is a hallmark of aging and various pathologies, including cancer, neurodegenerative disorders, and metabolic diseases, making strategies to modulate NAD levels a promising therapeutic frontier. This review provides the first integrated overview of NAD concentrations across cellular compartments (cytosol, mitochondria, nucleus, endoplasmic reticulum, Golgi, peroxisomes, and the extracellular space) together with measurement and modulation strategies. We summarize current knowledge on NAD distribution within organelles, address key challenges in accurate quantification, and highlight established and emerging approaches for both global and compartment-specific analysis. Finally, we discuss therapeutic strategies, from NAD⁺ precursor supplementation to enzyme modulators and gene therapy, highlighting both their translational potential and current limitations in treating diverse diseases and prolonging life and health span. Full article

(This article belongs to the Section Cell Metabolism)

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10 pages, 235 KB

Open AccessEditor’s ChoiceArticle

Exploratory Associations of Inflammatory Cytokines, Brain-Derived Neurotrophic Factor, and Vascular Endothelial Growth Factor with Clinical Outcomes in Patients with Bipolar Disorder

by Fumito Hamada, Leo Gotoh, Yuko Tomiyama, Hiroko Sugawara, Muneaki Ogata, Hiroki Kumagai, Ryo Asada, Ryusei Hatae, Kiyohiro Yasumatsu and Hikaru Hori

Metabolites 2026, 16(5), 328; https://doi.org/10.3390/metabo16050328 - 14 May 2026

Viewed by 672

Abstract

Background/Objectives: Bipolar disorder is characterized by psychosocial dysfunction, cognitive impairment, and incomplete recovery. Although inflammatory and neurotrophic mechanisms have been implicated, their relationships with multidimensional recovery outcomes remain unclear. We examined the relationships of inflammatory cytokines, brain-derived neurotrophic factor (BDNF), and vascular endothelial [...] Read more.

Background/Objectives: Bipolar disorder is characterized by psychosocial dysfunction, cognitive impairment, and incomplete recovery. Although inflammatory and neurotrophic mechanisms have been implicated, their relationships with multidimensional recovery outcomes remain unclear. We examined the relationships of inflammatory cytokines, brain-derived neurotrophic factor (BDNF), and vascular endothelial growth factor (VEGF) with depressive symptoms, psychosocial functioning, cognitive performance, personal recovery, and quality of life (QOL) in patients with bipolar disorder. Methods: This cross-sectional study of 24 patients with bipolar disorder assessed depressive symptoms, psychosocial functioning, cognitive functions, personal recovery, and QOL. Plasma tumor necrosis factor alpha, interleukin (IL)-6, IL-1β, IL-2, BDNF, and VEGF-A were measured by assay. Results: Subjective cognitive dysfunction was significantly associated with depressive symptom severity (rho = 0.53, p = 0.0083) and reduced QOL (rho = −0.56, p = 0.0042). Depressive symptoms were also associated with lower WHO-QOL-26 scores (rho = −0.43, p = 0.038). Significant interrelationships were observed among objective cognitive measures, and after false discovery rate (FDR) correction, the associations between FAST and PDQ-5-D, Symbol Check and Codebreaker, and Codebreaker and Trail remained statistically significant. High plasma IL-6 levels were associated with worse executive function (rho = 0.43, p = 0.0068). Higher VEGF levels were associated with better attentional performance (rho = −0.42, p = 0.042). Plasma IL-1β levels were positively associated with QOL (rho = 0.54, p = 0.02). After FDR correction, only the association between IL-1β and QOL remained statistically significant. Conclusions: This pilot study suggests that there may be associations between cognitive impairment and cytokines, as well as between quality of life and VEGF, in bipolar disorder. Further studies with larger sample sizes are needed. Full article

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

23 pages, 9939 KB

Open AccessEditor’s ChoiceArticle

Extraction Temperatures Shape Water-Soluble Metabolite Profiles of Nepeta nuda L. and thus Modulate the Bioactive Properties

by Desislava Mantovska, Alexandra Kapogianni, Ginka Cholakova, Ivanka Tsacheva, Anton Hinkov, Detelina Petrova, Zlatina Gospodinova, Georgi Antov, Danijela Mišić, Krasimir Rusanov, Mila Rusanova, Kalina Shishkova, Momchil Paunov, Zhenya Yordanova and Miroslava Zhiponova

Metabolites 2026, 16(5), 323; https://doi.org/10.3390/metabo16050323 - 13 May 2026

Viewed by 809

Abstract

Background: Plants of the genus Nepeta are widely used in ethnomedicine for treating inflammatory disorders due to their rich content of bioactive compounds. This study investigated how extraction temperature specifically affects the bioactive potential of aqueous extracts from wild-grown Nepeta nuda L. Methods: [...] Read more.

Background: Plants of the genus Nepeta are widely used in ethnomedicine for treating inflammatory disorders due to their rich content of bioactive compounds. This study investigated how extraction temperature specifically affects the bioactive potential of aqueous extracts from wild-grown Nepeta nuda L. Methods: The previously used maceration approach for this plant was applied at 30–60 °C to flowers, leaves, and stems. Phytochemical profiling included spectrophotometric assays, metabolite identification, and quantification. Biological activities reported for this plant were assessed, including antioxidant, anti-inflammatory, antiviral, antiproliferative, and antibacterial capacities. Results: Extraction yield was highest in flowers and leaves, where it increased significantly with rising temperature, while stems were less productive. All plant organs exhibited notable bioactivity falling into two groups: lower temperatures (30 and 40 °C) were optimal for antiviral and anti-inflammatory effects, whereas and higher temperatures (50 and 60 °C) enhanced antioxidant potential. The phytochemical composition, evaluated at representative extraction temperatures, revealed differential accumulation of p-coumaric acid and luteolin in all organs at 40 °C, while extraction at 60 °C corresponded to elevated levels of phenolic compounds. Flower extracts were confirmed to have the richest metabolic composition and were therefore subjected to further investigation. Extracts obtained at 40 °C influenced C1q binding, supporting their anti-inflammatory activity, whereas extraction at 60 °C resulted in stronger antiproliferative activity in colon cancer cell line. Antibacterial effects were similar at both temperatures. Conclusions: These findings highlight the importance of optimizing extraction conditions for future pharmacological applications of N. nuda. Full article

(This article belongs to the Special Issue Bioactive Metabolites from Natural Sources (2nd Edition))

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27 pages, 6945 KB

Open AccessEditor’s ChoiceArticle

Targeting the Gut–Brain Axis: Protective Effects of NMN in Alleviating D-Galactose-Induced Cognitive Deficits

by Zhenyang Zang, Feng Chen, Qiulian Tang, Wang Luo, Yuxian Lin, Jianxin Li and Yingcong Yu

Metabolites 2026, 16(5), 314; https://doi.org/10.3390/metabo16050314 - 6 May 2026

Viewed by 1043

Abstract

Background: To evaluate the neuroprotective effect of nicotinamide mononucleotide (NMN), an NAD⁺ precursor, in a D-galactose-induced aging mouse model. Chronic D-galactose administration is widely used to establish age-related cognitive impairment driven by oxidative stress. Methods: Mice received subcutaneous D-galactose for [...] Read more.

Background: To evaluate the neuroprotective effect of nicotinamide mononucleotide (NMN), an NAD⁺ precursor, in a D-galactose-induced aging mouse model. Chronic D-galactose administration is widely used to establish age-related cognitive impairment driven by oxidative stress. Methods: Mice received subcutaneous D-galactose for six weeks, concomitantly with oral NMN (300 or 500 mg/kg). Cognitive function was assessed using the Y-maze test and the Elevated Plus Maze test. Oxidative stress indicators, inflammatory cytokines, and Nrf2/HO-1 pathway components were measured by ELISA, Western blotting, and Immunohistochemistry. Gut microbiota composition was analyzed via 16S rRNA sequencing. Results: NMN supplementation improved spatial memory without affecting anxiety-related behavior. NMN enhanced the activities of antioxidant enzymes (SOD, GSH, CAT), reduced malondialdehyde and pro-inflammatory cytokine levels and decreased microglial activation in the hippocampus. Furthermore, NMN remodeled the gut microbiota by increasing butyrate-producing taxa (such as Butyrivibrio_A and Clostridium_T) and activated the Nrf2/HO-1 signaling pathway. Conclusions: NMN alleviates age-related cognitive decline in mice by reducing oxidative stress, suppressing neuroinflammation, and modulating the gut microbiota. Targeting the gut–brain axis and the Nrf2/HO-1 pathway may therefore represent a promising therapeutic strategy for age-related neurodegeneration. Full article

(This article belongs to the Special Issue Microbial Metabolites and Host Health)

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22 pages, 18258 KB

Open AccessEditor’s ChoiceArticle

Physiology, Metabolomics, and Transcriptomics Reveal Effects of AMF and Chaetomium globosum Co-Inoculation on Growth and Medicinal Compounds in Astragalus membranaceus

by Yuxin Gong, Shengnan Liu, Xiaoju Zhao, Weisan Zhang, Jiaotong Li, Mengqi Liu, Xueqi Zhang, Hanqi Jia and Zhonghua Tang

Metabolites 2026, 16(5), 313; https://doi.org/10.3390/metabo16050313 - 3 May 2026

Viewed by 626

Abstract

Background/Objectives: Microbial inoculants effectively alleviate inhibitory factors during plant cultivation; however, the effects and underlying mechanisms of arbuscular mycorrhizal fungi (AMF) and Chaetomium globosum on the growth, metabolism, and bioactive compound production of A. membranaceus are still poorly understood. Methods: In this [...] Read more.

Background/Objectives: Microbial inoculants effectively alleviate inhibitory factors during plant cultivation; however, the effects and underlying mechanisms of arbuscular mycorrhizal fungi (AMF) and Chaetomium globosum on the growth, metabolism, and bioactive compound production of A. membranaceus are still poorly understood. Methods: In this experiment, different concentrations of C. globosum (10⁴, 10⁶, 10⁸ spores/mL) (Q1, Q2, Q3), AMF, and their combined treatments (AQ1, AQ2, AQ3) were applied to A. membranaceus seedlings via root irrigation, with an equal amount of sterile water as a control (CK). Results: The results showed that: (1) Under single inoculation with C. globosum, root colonization rate increased with higher inoculation concentrations, reaching its peak at Q3. Additionally, AQ3 significantly enhanced AMF colonization in A. membranaceus, and the presence of C. globosum promoted AMF root colonization and expansion. (2) AQ3 significantly enhanced the growth and photosynthesis of A. membranaceus, while also demonstrating excellent efficacy in alleviating lipid peroxidation-induced damage. (3) AQ3 treatment led to increased accumulation of major bioactive compounds in A. membranaceus, including calycosin-7-glucoside, cycloastragenol, and astragalosides I–III. (4) AQ3 treatment significantly upregulated multiple key structural genes involved in phenylalanine metabolism and flavonoid biosynthesis pathways, including PAL, 4CL, FLS, and CYP75B1. Conclusions: This upregulation enhanced the metabolic flux allocation from L-phenylalanine toward downstream flavonoid metabolites, thereby promoting the accumulation of major flavonoid constituents of A. membranaceus, such as galangin and luteolin, in both roots and leaves. Full article

(This article belongs to the Special Issue Bioactive Metabolites from Fungal Endophytes Associated with Plants)

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17 pages, 5036 KB

Open AccessEditor’s ChoiceArticle

HR-LCMS/MS-Based Dereplication of Plant-Derived Autophagy Inducers Revealed Astragalus dasyanthus as a New Glabrol Producer

by Anastasiia K. Bolikhova, Vera A. Alferova, Anton R. Izzi, Arina A. Nikandrova, Gulnara K. Kudryakova, Mikhail Y. Zhitlov, Ekaterina A. Guseva, Dmitry S. Karpov, Dmitrii A. Lukianov, Olga A. Dontsova and Petr V. Sergiev

Metabolites 2026, 16(5), 311; https://doi.org/10.3390/metabo16050311 - 1 May 2026

Viewed by 863

Abstract

Background/Objectives: Autophagy is an important cellular self-cleansing process whose normal functioning is essential for preventing many age-related diseases. The search for and study of new autophagy activators allows the proposal of novel therapeutic approaches for the treatment of age-related diseases. Medical plants are [...] Read more.

Background/Objectives: Autophagy is an important cellular self-cleansing process whose normal functioning is essential for preventing many age-related diseases. The search for and study of new autophagy activators allows the proposal of novel therapeutic approaches for the treatment of age-related diseases. Medical plants are a rich source of bioactive compounds with variable functions. In this study, we propose an HR-LCMS/MS-based technique for identifying the principal autophagy activators in plant extracts. Methods: We performed a Western blot analysis of the autophagy-inducing activity of plant extract HPLC fractions on a model of SH-SY5Y cells. The composition of the fractions showing autophagy-activating potential was determined via HR-LCMS/MS. Results: We analyzed five plants known to produce autophagy activators and proved the ability of the method to detect the main bioactive compounds. Additional screening demonstrated for the first time that Astragalus dasyanthus is a producer of the autophagy-inducer glabrol. Conclusions: The described HR-LCMS/MS-based method for identifying autophagy activators in multicomponent plant extracts is effective and could be used for further high-throughput screening. Full article

(This article belongs to the Special Issue LC-MS/MS Analysis for Plant Secondary Metabolites, 2nd Edition)

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12 pages, 938 KB

Open AccessEditor’s ChoiceArticle

Circulating Wnt Signaling Inhibitors and Osteoprotegerin in Women with Newly Diagnosed Overt Thyroid Dysfunction

by Mariya Zhivkova Miteva, Maria Mitkova Orbetzova, Boyan Ivanov Nonchev, Delyana Miteva Davcheva and Kostadin Gigov

Metabolites 2026, 16(5), 308; https://doi.org/10.3390/metabo16050308 - 30 Apr 2026

Viewed by 498

Abstract

Background: Thyroid hormones influence bone metabolism, and autoimmune thyroid diseases may further impact skeletal homeostasis. Wnt signaling inhibitors, including Dickkopf-1 (DKK-1) and sclerostin (SOST), as well as osteoprotegerin (OPG), play key roles in regulating bone formation and resorption. This study aimed to [...] Read more.

Background: Thyroid hormones influence bone metabolism, and autoimmune thyroid diseases may further impact skeletal homeostasis. Wnt signaling inhibitors, including Dickkopf-1 (DKK-1) and sclerostin (SOST), as well as osteoprotegerin (OPG), play key roles in regulating bone formation and resorption. This study aimed to evaluate circulating DKK-1, SOST, and OPG in women with newly diagnosed overt thyroid dysfunction. Methods: This cross-sectional study included 62 women with newly diagnosed, untreated overt thyroid dysfunction (35 hypothyroid and 27 hyperthyroid) and 33 age- and BMI-matched healthy controls. Serum levels of DKK-1, sclerostin, and OPG were measured using ELISA. Thyroid function and autoantibodies were assessed using automated immunoassays. Correlation analysis was performed to evaluate associations between variables. Results: Serum DKK-1 levels were significantly elevated in both hypothyroid and hyperthyroid women compared with controls (p < 0.001). Sclerostin levels showed a non-significant trend toward higher values. OPG levels were significantly increased in hyperthyroid patients and moderately elevated in hypothyroid patients. Significant positive correlations were observed between OPG and FT3 (r = 0.42, p = 0.001) and FT4 (r = 0.43, p = 0.001). In hypothyroid patients, OPG correlated positively with TgAb (r = 0.46, p = 0.007). A strong positive correlation was found between DKK-1 and SOST (p < 0.001), while DKK-1 was negatively associated with age (p < 0.05). Conclusions: Overt thyroid dysfunction is associated with significant alterations in circulating Wnt signaling inhibitors and OPG. These findings suggest a potential role of Wnt signaling and immune–bone interactions in thyroid-related changes in bone metabolism. Full article

(This article belongs to the Special Issue Exploring Metabolomic Signatures and the Metabolic Impact of Thyroid Dysfunction)

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18 pages, 1911 KB

Open AccessEditor’s ChoiceArticle

High-Resolution Magic Angle Spinning Metabolomic Profiling of IDH-Wild-Type Glioblastoma Reveals a Composite Surgical Sampling Signature Shaped by Clinical and Anatomical Tumor Features

by Julien Todeschi, Caroline Bund, Hassiba Outilaft, Hélène Cebula and Izzie-Jacques Namer

Metabolites 2026, 16(5), 296; https://doi.org/10.3390/metabo16050296 - 27 Apr 2026

Viewed by 524

Abstract

Background/Objectives: Tissue-based metabolomic readouts in IDH-wild-type glioblastoma may be strongly shaped by how tumor tissue is surgically accessed and sampled. We aimed to determine whether, and to what extent, surgical sampling context structures the HRMAS metabolic landscape, and to disentangle sampling-related contributions from [...] Read more.

Background/Objectives: Tissue-based metabolomic readouts in IDH-wild-type glioblastoma may be strongly shaped by how tumor tissue is surgically accessed and sampled. We aimed to determine whether, and to what extent, surgical sampling context structures the HRMAS metabolic landscape, and to disentangle sampling-related contributions from clinico-anatomical confounders. Methods: We retrospectively analyzed 99 patients with de novo IDH-wild-type glioblastoma (35 biopsy-only, 64 resection: 40 gross-total, 21 near-total, 3 subtotal), yielding 166 HRMAS spectra and 47 quantified metabolites (nmol/mg). Patient-level profiles were compared using PCA, metabolite-wise testing, pathway-level aggregation (10 pathways), and variance partitioning by PERMANOVA, both unadjusted and adjusted for age, WHO PS, deep-seated location, midline involvement, multifocality, MGMT methylation, and eloquent area. Sensitivity analyses included clinico-anatomically restricted subgroups, 15 canonical metabolite ratios, and Probabilistic Quotient Normalization. Intratumoral heterogeneity was assessed in 44 multi-sampled patients. Results: Biopsy-only and resection-derived cases separated along PC1 in unsupervised PCA (62.6% variance; p < 0.001), with 42/47 metabolites differing after FDR correction. However, the surgical group explained only 2.6% of the total variance (PERMANOVA p = 0.026), and this share was no longer significant after confounder adjustment (p = 0.39). Clinico-anatomical restriction progressively attenuated the effect (42/47 → 1/47 significant metabolites). Ratio-based and PQN analyses showed a residual compositional difference beyond scaling (13/15 ratios; 16/47 metabolites). Intratumoral heterogeneity was greater in resections and preserved in an n-matched analysis (p = 0.020). Conclusions: The apparent biopsy-versus-resection metabolic difference is largely a composite signal reflecting clinico-anatomical patient selection with a smaller tissue-composition contribution. Biopsy-only and resection-derived specimens should not be pooled uncritically in tissue-based metabolomic studies of glioblastoma. Full article

(This article belongs to the Special Issue Recent Advances in Metabolomics (IECM2026))

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11 pages, 446 KB

Open AccessEditor’s ChoiceArticle

The Hormonal–Metabolic Puzzle of PCOS: Linking AMH Levels, Body Fat Distribution, and Insulin Resistance in Overweight and Obese Women

by Amalia Gorzko, Jolanta Nawrocka-Rutkowska, Agnieszka Brodowska, Edyta Śliwak, Andrzej Starczewski and Iwona Szydłowska

Metabolites 2026, 16(5), 295; https://doi.org/10.3390/metabo16050295 - 27 Apr 2026

Viewed by 630

Abstract

Background: The relationship between AMH (anti-Müllerian hormone) levels, fat distribution, and insulin resistance in women with PCOS has been widely studied, yet findings remain inconsistent. Recent guidelines emphasize the growing role of AMH in PCOS diagnosis and suggest its potential inclusion among [...] Read more.

Background: The relationship between AMH (anti-Müllerian hormone) levels, fat distribution, and insulin resistance in women with PCOS has been widely studied, yet findings remain inconsistent. Recent guidelines emphasize the growing role of AMH in PCOS diagnosis and suggest its potential inclusion among diagnostic criteria, highlighting its relevance for guiding therapeutic management. Objectives: This retrospective study aimed to evaluate the association between AMH levels and metabolic parameters in overweight and obese reproductive-age women with PCOS. Ethical approval was obtained from the bioethics committee. Methods: Two groups of 52 women each were selected from PCOS patients treated at our clinic between 2024 and 2025: one with a waist-to-hip ratio (WHR) ≤ 0.85 and the other with a WHR > 0.85. Venous blood samples were collected to measure AMH, fasting glucose, and fasting insulin. Body height and weight were recorded to calculate body mass index (BMI), and insulin resistance was assessed using HOMA-IR. Waist and hip circumferences were measured to determine WHR. Results: The association between central adiposity and AMH in overweight and obese women with PCOS depended on insulin resistance. In insulin-resistant women, higher WHR was linked to lower AMH, whereas in women without insulin resistance, higher WHR corresponded to higher AMH levels. Conclusions: Insulin resistance appears to influence AMH in opposite directions depending on a woman’s WHR, suggesting its potential role in tailoring individualized treatment strategies. Full article

(This article belongs to the Special Issue The Influencing Factors of Metabolic Changes in Polycystic Ovary Syndrome)

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16 pages, 2130 KB

Open AccessEditor’s ChoiceArticle

Toxicokinetic Studies of the Two Stimulants M-ALPHA and N-Methyl-cyclazodone Using In Vitro and In Vivo Tools

by Tanja M. Gampfer, Samira Klaes, Niels Eckstein and Markus R. Meyer

Metabolites 2026, 16(5), 291; https://doi.org/10.3390/metabo16050291 - 23 Apr 2026

Viewed by 2490

Abstract

Background/Objectives: Synthetic stimulants represent the most prevalent subclass on the new psychoactive substances (NPSs) market. However, the toxicokinetic properties of M-ALPHA, a regioisomer of MDMA and N-methyl-cyclazodone a pemoline derivative, are not yet characterized. Methods: Therefore, this study investigated the metabolism of [...] Read more.

Background/Objectives: Synthetic stimulants represent the most prevalent subclass on the new psychoactive substances (NPSs) market. However, the toxicokinetic properties of M-ALPHA, a regioisomer of MDMA and N-methyl-cyclazodone a pemoline derivative, are not yet characterized. Methods: Therefore, this study investigated the metabolism of both NPSs in pooled liver S9 fraction and rat urine, characterized cytochrome P450 (CYP) kinetics and plasma protein binding (PPB), and assessed the CYP inhibition potential of M-ALPHA, using high-performance liquid chromatography coupled to high resolution tandem mass spectrometry (HPLC-HRMS/MS). Results: Four metabolites of M-ALPHA were detected including one phase I and three phase II metabolites, resulting from demethylenation followed by subsequent methylation or glucuronidation. For N-methyl-cyclazodone, one phase I metabolite formed via N-demethylation was identified. The primary enzymes involved in M-ALPHA metabolism were CYP2B6 and CYP2D6. Notably, M-ALPHA inhibited these enzymes to a strong or moderate extent, respectively. In contrast, the metabolism of N-methyl-cyclazodone was primarily mediated by CYP2A6. PPB studies indicated low-to-moderate binding for both compounds, suggesting that significant protein-binding interactions are unlikely. Conclusions: As M-ALPHA only formed metabolites that overlapped with those of MDMA, differing only by minor retention time shifts, reliable HPLC-HRMS/MS-based identification may be challenging in clinical and forensic toxicology settings as well as doping analysis. Furthermore, drug–drug interactions following polydrug use cannot be excluded for either NPS, particularly when co-ingested with other CYP substrates metabolized by the same isoforms. Full article

(This article belongs to the Special Issue Metabolite Profiling of Novel Psychoactive Substances)

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12 pages, 443 KB

Open AccessEditor’s ChoiceArticle

Atherogenic Index of Plasma Relationship with Cardiovascular Risk Factors and Frailty and Value as Determinant of Mortality in Elderly Patients with Severe Aortic Stenosis

by Annamaria Mazzone, Melania Gaggini and Cristina Vassalle

Metabolites 2026, 16(5), 289; https://doi.org/10.3390/metabo16050289 - 22 Apr 2026

Viewed by 564

Abstract

Background: Frailty is a common finding in elderly subjects with severe aortic stenosis (AoS) and a strong predictor of mortality and disability after aortic valve surgery. The atherogenic index of plasma (AIP) is related to different cardiovascular (CV) risk factors, which in [...] Read more.

Background: Frailty is a common finding in elderly subjects with severe aortic stenosis (AoS) and a strong predictor of mortality and disability after aortic valve surgery. The atherogenic index of plasma (AIP) is related to different cardiovascular (CV) risk factors, which in turn are correlated to the progression of frailty as well as of AoS. Aim: to analyze the association of AIP with different CV risk factors and frailty scores and its value as a determinant of mortality in older adults with severe AoS. Methods: The association of AIP with a multidimensional assessment of frailty by using Fried criteria and the following indices; timed up-and-go test (TUG) for gait function; Charlson Index (CI), basic activities of daily living (BADL) and instrumental activities of daily living (IADL) for disability; mini–mental state examination for cognitive function evaluation (MMSE); Geriatric Depression Score for mood disorder (GDS); Mini Nutritional Assessment (MNA) for nutritional status was assessed in 102 elderly AoS patients (33 males; mean age 83 ± 6 yrs). Moreover, the relationship between AIP and demographic, lifestyle, traditional CV risk factors and CV mortality was also evaluated. Results: Significant relationships between AIP and glycemia and inflammatory parameters (CRP, ESR and fibrinogen) as well as with troponin I were found. Moreover, AIP significantly correlates with CI, BADL, IADL and MNA. However, the Kaplan–Meier analysis did not show any significant difference for survival rates according to AIP intervals of risk, whereas ejection fraction remained the only significant determinant after multivariate adjustment for mortality at the Cox proportional hazard models analysis in this patient population. Conclusions: Higher AIP is significantly associated with cardiometabolic risk and increased physical dysfunction risk and frailty in AoS pts, evidencing its potential use as a simple biomarker in this clinical setting, although it did not represent a significant determinant for mortality in this population. Full article

(This article belongs to the Special Issue Lipid Metabolism in Age-Related Diseases: 2nd Edition)

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18 pages, 1672 KB

Open AccessEditor’s ChoiceReview

A Structured Computational Roadmap for Lipidomics in R: Reproducible Workflows from Raw Data to Functional Insight

by Maria-Christina P. Papatheodorou, Panagiotis Vlamos and Marios G. Krokidis

Metabolites 2026, 16(5), 288; https://doi.org/10.3390/metabo16050288 - 22 Apr 2026

Viewed by 1286

Abstract

Lipidomics has emerged as a transformative discipline in biomedical research, providing high-resolution insights into metabolic signaling and disease pathophysiology. The R programming language provides a widely adopted framework for extensible analysis of complex lipidomic datasets due to its robust biostatistical infrastructure. Herein, we [...] Read more.

Lipidomics has emerged as a transformative discipline in biomedical research, providing high-resolution insights into metabolic signaling and disease pathophysiology. The R programming language provides a widely adopted framework for extensible analysis of complex lipidomic datasets due to its robust biostatistical infrastructure. Herein, we present a comprehensive roadmap for lipidomics in R, structured around a standardized analytical lifecycle: from raw data acquisition and preprocessing to structural annotation, statistical modeling and functional interpretation. We critically contextualize and integrate a curated suite of widely adopted R packages (version 4.3.0), including xcms and MSnbase for feature extraction, LipidMS 3.0 for fragmentation-based identification, and lipidr for quality control and normalization. Furthermore, we demonstrate how advanced tools such as mixOmics and clusterProfiler can be integrated to bridge the gap between differential lipid abundance and systems-level biological insights. Particular emphasis is placed on reproducibility, nomenclature standardization and the emerging role of machine learning in biomarker discovery. By synthesizing these resources into a coherent pipeline, this guide provides a structured reference for researchers. Further discussion addresses methodological pitfalls, statistical assumptions and reproducibility constraints that frequently compromise lipidomics studies. Ultimately, this structured approach facilitates systematic tool selection, accelerating the translation of complex lipidomic signatures into reproducible and clinically meaningful discoveries. Full article

(This article belongs to the Special Issue Lipidomic and Metabolomic Analysis of Neurodegenerative Diseases)

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17 pages, 2662 KB

Open AccessEditor’s ChoiceArticle

Effects of a Reprometabolic Syndrome-Inducing Eucaloric High-Fat Diet on Insulin Sensitivity, Body Composition, the Lipidome, and the Microbiome

by Irene E. Schauer, Katherine Kuhn, Andrew P. Bradford, Angela J. Fought, Daniel N. Frank, Cassandra V. Kotter, Charles E. Robertson, Katie Duffy and Nanette Santoro

Metabolites 2026, 16(5), 286; https://doi.org/10.3390/metabo16050286 - 22 Apr 2026

Viewed by 907

Abstract

Background: We previously demonstrated recapitulation of the relative hypogonadotropic hypogonadism of obesity, the Reprometabolic Syndrome (RMS), in women of normal BMI with a one-month high-fat, eucaloric diet (HFD). Objective: Assess effects of HFD on sleep, body composition and lifestyle and metabolic [...] Read more.

Background: We previously demonstrated recapitulation of the relative hypogonadotropic hypogonadism of obesity, the Reprometabolic Syndrome (RMS), in women of normal BMI with a one-month high-fat, eucaloric diet (HFD). Objective: Assess effects of HFD on sleep, body composition and lifestyle and metabolic secondary outcomes and correlate insulin sensitivity changes with the RMS. Methods: A total of 18 normally cycling women aged 18–38 with BMI 18–24 kg/m² were enrolled for a four-month study including a eucaloric HFD (48% calories from fat) for one menstrual cycle. Activity, sleep, body composition, and the lipidome were measured in all participants. Fecal microbiome was analyzed in the last nine participants, and insulin sensitivity by two-stage hyperinsulinemic euglycemic clamp was measured before and after HFD in 15 participants. Results: Relative to the pre-diet period, BMI, activity and sleep measures did not change, except for waking after sleep onset (WASO), which appeared to decrease during and post HFD. DXA revealed statistically significant decreases in total percent fat, total fat mass, visceral fat volume, and trunk fat volume. Whole-body insulin sensitivity decreased with the HFD while adipocyte insulin sensitivity was unaffected. Insulin sensitivity changes did not correlate with change in gonadotropins or response to gonadotropin releasing hormone (GnRH). Multiple significant changes in plasma lipids were observed, including increased ceramides and glucosylceramides. Microbiome analysis revealed increased microbial diversity. Conclusions: A one-month eucaloric HFD that induced RMS in normal-weight, reproductive-aged women also induced whole-body insulin resistance (IR) and multiple lipidomics changes potentially associated with IR. These changes in IR occurred despite overall stable activity, BMI and sleep, but did not correlate with the HPO axis defects. The unexpected decrease in body fat and increase in microbial diversity may be related to specific dietary elements of the HFD. Full article

(This article belongs to the Special Issue Effects of Nutrition Intake and Lifestyle Intervention on Human Metabolic Health)

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15 pages, 659 KB

Open AccessEditor’s ChoiceReview

Altered Lipid Metabolism in Psoriatic Arthritis: A Comprehensive Review

by Stanislava Popova-Belova, Mariela Geneva-Popova, Stefka Stoilova, Velichka Popova, Georgi Nikolov and Dimitar Nikolov

Metabolites 2026, 16(5), 287; https://doi.org/10.3390/metabo16050287 - 22 Apr 2026

Viewed by 715

Abstract

Psoriatic arthritis (PsA) is a chronic inflammatory disorder affecting both the joints and skin. Beyond musculoskeletal manifestations, patients with PsA frequently exhibit alterations in lipid metabolism, contributing to an increased risk of cardiovascular disease. Dyslipidemia in PsA arises from multiple mechanisms, including systemic [...] Read more.

Psoriatic arthritis (PsA) is a chronic inflammatory disorder affecting both the joints and skin. Beyond musculoskeletal manifestations, patients with PsA frequently exhibit alterations in lipid metabolism, contributing to an increased risk of cardiovascular disease. Dyslipidemia in PsA arises from multiple mechanisms, including systemic inflammation, insulin resistance, and imbalances in adipokines such as leptin, adiponectin, and resistin. A structured literature search was conducted in PubMed, Scopus, and Web of Science to identify relevant studies on lipid metabolism in psoriatic arthritis, and the evidence was synthesized narratively. PsA is also commonly associated with obesity and metabolic syndrome, further exacerbating dyslipidemia and cardiovascular risk. Interventions including weight loss, lifestyle modification, and anti-inflammatory treatments have been shown to improve lipid profiles and clinical outcomes. This review provides a comprehensive overview of current knowledge on altered lipid metabolism in PsA, highlighting underlying mechanisms, clinical implications, and therapeutic strategies to reduce cardiovascular risk. Full article

(This article belongs to the Special Issue Psoriasis and Metabolic Syndrome)

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13 pages, 703 KB

Open AccessEditor’s ChoiceArticle

Comparative Analysis of Two Dietary Saturated Fat Types on Metabolite Profiles Crossing the Blood–Brain Barrier of Poultry Chicks

by Oluteru E. Orimaye, Paul C. Omaliko, Nathanael I. Lichti, Bruce R. Cooper and Yewande O. Fasina

Metabolites 2026, 16(4), 283; https://doi.org/10.3390/metabo16040283 - 20 Apr 2026

Viewed by 737

Abstract

Background: The dorsal raphe nucleus (DRN) produces and distributes serotonin, while the hypothalamus (HYP) uses serotonergic signals to regulate physiological processes in chickens. Coconut oil (COCO), rich in medium-chain fatty acids, is rapidly absorbed without re-esterification. Methods: Day-old broilers (Ross 708 male, n [...] Read more.

Background: The dorsal raphe nucleus (DRN) produces and distributes serotonin, while the hypothalamus (HYP) uses serotonergic signals to regulate physiological processes in chickens. Coconut oil (COCO), rich in medium-chain fatty acids, is rapidly absorbed without re-esterification. Methods: Day-old broilers (Ross 708 male, n = 160) were distributed into two dietary treatments with five replicates of 16 birds each. The birds were fed a corn–soybean meal (SBM) basal diet supplemented with 3% of poultry fat (CON) or coconut oil (COCO). The body-weight gain (BWG), feed intake (FI), and feed conversion ratio (FCR) were recorded over a 3-week period, and the data were subjected to a t-test. Untargeted metabolomic analysis by high-performance liquid chromatography (HPLC-MS) was used to evaluate the influence of the type of dietary fat on metabolite profiles in the DRN, HYP, and plasma of broiler chickens. Principal component analysis (PCA) was used to identify unique metabolites, and ANOVA was used to identify the metabolites that were significantly abundant (p < 0.05). The metabolites were then annotated using the KEGG and HMDB databases. Results: Birds in the COCO treatment gained more weight on average (0.8446 kg/bird) than birds in the CON group (0.8132 kg/bird; p = 0.0496). Five metabolites associated with multiple significant cellular processes, such as brain function, energy metabolism, and neurotransmission, showed similar differential expression patterns, while two metabolic pathways (butanoate metabolism and alanine, aspartate and glutamate metabolism) were identified. Conclusions: The dietary inclusion of COCO improves BWG in poultry and enhances their overall well-being by modulating metabolite profiles, supporting neurotransmission, and enriching the metabolic pathways essential for growth and brain function. Full article

(This article belongs to the Section Metabolomic Profiling Technology)

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19 pages, 714 KB

Open AccessEditor’s ChoiceArticle

Red Blood Cell Distribution Width and Neutrophil-to-Lymphocyte Ratio as Markers of Cardiovascular Disease and Vascular Calcification in Chronic Kidney Disease: A Large Cohort Study

by Anastasios Zagaliotis, Athanasios Roumeliotis, Stefanos Roumeliotis, Ioannis E. Neofytou, Garyfallia Varouktsi, Eirini Leptokaridou-Mourtzila, Aikaterini Stamou, Vasiliki Sgouropoulou, Gordana Kocic, Andrej Veljkovic, Rudolf Bittner, Willi Jahnen-Dechent, Leon J. Schurgers and Vassilios Liakopoulos

Metabolites 2026, 16(4), 280; https://doi.org/10.3390/metabo16040280 - 20 Apr 2026

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Abstract

Background/Objectives: Cardiovascular disease (CVD) in chronic kidney disease (CKD) arises from a multifaceted interplay of pathophysiological processes, including chronic inflammation, oxidative stress (OS), and accelerated vascular calcification (VC). Red blood cell distribution width (RDW) and the neutrophil-to-lymphocyte ratio (NLR) have emerged as simple, [...] Read more.

Background/Objectives: Cardiovascular disease (CVD) in chronic kidney disease (CKD) arises from a multifaceted interplay of pathophysiological processes, including chronic inflammation, oxidative stress (OS), and accelerated vascular calcification (VC). Red blood cell distribution width (RDW) and the neutrophil-to-lymphocyte ratio (NLR) have emerged as simple, inexpensive, and readily available hematological indices that may capture these underlying disturbances. As such, they hold promise as accessible biomarkers for stratifying cardiovascular risk in patients with CKD. Methods: This cross-sectional study enrolled 497 patients, comprising 477 with CKD across all stages and 20 controls. We evaluated the associations of RDW and NLR with both traditional and non-traditional cardiovascular risk factors, as well as with serum calcification propensity (T50). Spearman’s correlation and multivariable regression analysis were used to assess these relationships. Results: Both RDW and NLR were significantly elevated in patients with established CVD (p < 0.001 for both) and demonstrated a progressive increase across advancing CKD stages (p < 0.001). RDW and NLR showed positive correlations with age, CVD duration, urea, phosphorus, parathormone, CRP, FG23, and mean carotid intima–media thickness (cIMT), while exhibiting inverse correlations with eGFR, serum albumin, hemoglobin, lipids, antioxidants such as superoxide dismutase, fetuin-A, and T50. Additionally, NLR correlated positively with the duration of hypertension and diabetes, as well as with albuminuria. Quartile analysis revealed a stepwise decline in T50 across increasing categories of RDW and NLR, supporting the link with impaired calcification defense. In multivariable analysis, T50 independently predicted NLR (β = −0.013; p = 0.018), whereas total cholesterol (β = −0.011; p = 0.019) and cIMT (β = 0.38; p = 0.018) emerged as independent determinants of RDW. Conclusions: RDW and NLR strongly reflect the burden of inflammation, metabolic disturbance, and vascular dysfunction in patients across the CKD spectrum. The consistent associations with impaired calcification defense and with established cardiovascular risk markers underscore the potential value as accessible indicators of cardiovascular vulnerability in CKD. These findings support incorporating RDW and NLR into routine risk assessment and highlight T50 as a mechanistically relevant determinant of hematologic inflammation profiles. Full article

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

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13 pages, 2754 KB

Open AccessEditor’s ChoiceArticle

Selected Brain Metabolites and Mitochondrial DNA Copy Number as Potential Markers of Ongoing Neurodegeneration in Patients with Wolfram Syndrome

by Ewa Zmysłowska-Polakowska, Tomasz Płoszaj, Sebastian Skoczylas, Julia Grzybowska-Adamowicz, Dobromiła Barańska, Katarzyna Matera, Aleksandra Palatyńska-Ulatowska, Wojciech Młynarski, Agnieszka Zmysłowska and Michal Ciborowski

Metabolites 2026, 16(4), 281; https://doi.org/10.3390/metabo16040281 - 20 Apr 2026

Viewed by 921

Abstract

Background: Wolfram syndrome (WFS) is a rare neurodegenerative disease that is genetically determined and inherited in an autosomal recessive manner. Although the first clinical symptom appearing in early childhood is diabetes mellitus, subsequent symptoms are associated with optic nerve atrophy, followed by [...] Read more.

Background: Wolfram syndrome (WFS) is a rare neurodegenerative disease that is genetically determined and inherited in an autosomal recessive manner. Although the first clinical symptom appearing in early childhood is diabetes mellitus, subsequent symptoms are associated with optic nerve atrophy, followed by central nervous system atrophy. Methods: The aim of the study was to analyse magnetic resonance images (MRI) of the brain in combination with single-voxel magnetic resonance spectroscopy (MRS) and to assess the copy number of mitochondrial DNA (mtDNA-CN) in 10 patients with WFS compared with a control group of 17 healthy individuals. Results: A significant decrease in the amount of selected metabolites was observed in WFS patients compared to controls in all assessed brain regions (pons, cerebellum, white matter, thalamus, and hippocampus). For three metabolites, Glutamate (Glu), Glutamate + Glutamine (Glx) and total N-acetylaspartate (TNAA), significant differences in concentrations were found between the study groups in almost all matrices evaluating specific areas of the brain (p < 0.011), with the exception of a trend toward reduced TNAA in the hippocampus (p = 0.065). In addition, patients with WFS had a significant decrease in the mitochondrial-to-nuclear DNA ratio compared to controls (p < 0.0003). Some metabolites, such as N-acetylaspartate and total N-acetylaspartate, showed strong correlations with specific regions of the visual pathway on MRI scans in patients with WFS. Conclusions: Selected brain metabolites and mtDNA-CN may become potential markers of WFS, and the results of this study may be used to define indicators for future therapeutic strategies. Full article

(This article belongs to the Special Issue Brain Metabolic Alterations in Neurodegenerative Diseases)

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17 pages, 5410 KB

Open AccessEditor’s ChoiceArticle

Bile and Serum Metabolomics in Living Donor Liver Transplantation: Exploratory Insights into Acute Rejection Biomarkers

by Yuta Hirata, Yasunaru Sakuma, Hideo Ogiso, Taiichi Wakiya, Takahiko Omameuda, Toshio Horiuchi, Noriki Okada, Yukihiro Sanada, Yasuharu Onishi, Hironori Yamaguchi, Ryozo Nagai and Kenichi Aizawa

Metabolites 2026, 16(4), 273; https://doi.org/10.3390/metabo16040273 - 17 Apr 2026

Viewed by 1062

Abstract

Background: Acute rejection remains a major complication following liver transplantation, yet reliable noninvasive biomarkers for its early prediction and diagnosis remain unidentified. This exploratory study characterized bile and serum metabolites associated with acute rejection in living donor liver transplantation using comprehensive metabolomic profiling [...] Read more.

Background: Acute rejection remains a major complication following liver transplantation, yet reliable noninvasive biomarkers for its early prediction and diagnosis remain unidentified. This exploratory study characterized bile and serum metabolites associated with acute rejection in living donor liver transplantation using comprehensive metabolomic profiling combined with machine learning. Methods: Non-targeted metabolomics were performed on bile samples collected on post-operative day (POD) 1 (n = 38) and serum on POD 14 (n = 45) from liver transplant recipients. Partial least squares discriminant analysis-based variable selection was followed by logistic regression and least absolute shrinkage and selection operator models, which were evaluated via cross-validation in the discovery cohort to explore potential biomarkers for acute rejection. Results: A three-variable, bile-based model for predicting acute rejection achieved a mean cross-validated AUC of 0.872 (95% confidence interval: 0.814–0.930). Glycohyocholic acid and sulfolithocholylglycine were the main contributors. A nine-variable serum model for the Rejection Activity Index, including the change in γ-glutamyl transferase, showed a mean cross-validated R² of 0.728 (95% confidence interval: 0.609–0.846), with methionine, creatine, and oxidized fatty acids contributing prominently. Conclusions: These findings suggest that metabolomic profiling combined with machine learning may provide candidate biomarkers for acute rejection after liver transplantation. However, given the exploratory nature of the study and the lack of external validation, the clinical utility of these metabolite signatures remains to be determined. Therefore, external validation in larger, independent cohorts will be required. Full article

(This article belongs to the Special Issue Proteomics and Metabolomics in Human Health and Disease)

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12 pages, 231 KB

Open AccessEditor’s ChoiceArticle

Association Between Vitamin D Deficiency and Glycemic, Lipid, and Adiposity Markers in Older Adults: A Nationally Representative Study

by Yong-Joon Kim and Kyeongmin Jang

Metabolites 2026, 16(4), 270; https://doi.org/10.3390/metabo16040270 - 16 Apr 2026

Viewed by 794

Abstract

Background/Objectives: Vitamin D plays an important role in glucose metabolism, lipid regulation, and inflammatory processes, and has been implicated in cardiometabolic health. However, its associations with specific metabolic biomarkers remain inconsistent, particularly in older adults. This study aimed to examine whether vitamin D [...] Read more.

Background/Objectives: Vitamin D plays an important role in glucose metabolism, lipid regulation, and inflammatory processes, and has been implicated in cardiometabolic health. However, its associations with specific metabolic biomarkers remain inconsistent, particularly in older adults. This study aimed to examine whether vitamin D deficiency is differentially associated with multiple metabolic biomarkers in a nationally representative sample of older adults. Methods: This cross-sectional study used data from the 2024 Korea National Health and Nutrition Examination Survey, including 1806 adults aged ≥65 years. Vitamin D deficiency was defined as serum 25-hydroxyvitamin D levels < 20 ng/mL. Metabolic biomarkers included fasting glucose, glycated hemoglobin (HbA1c), triglycerides, C-reactive protein (CRP), high-density lipoprotein cholesterol (HDL-C), waist circumference, and body mass index (BMI). Complex sample linear regression analyses were performed with sequential adjustment for sociodemographic factors, health behaviors, and comorbidities. Results: In unadjusted analyses, vitamin D deficiency was associated with adverse metabolic profiles, including higher fasting glucose, HbA1c, triglycerides, waist circumference, and CRP levels, and lower HDL-C levels. After adjustment for sociodemographic factors, health behaviors, and comorbidities, significant associations remained for HbA1c (β = 0.10, p = 0.034), triglycerides (β = 0.10, p = 0.003), and waist circumference (β = 1.21, p = 0.040). No significant associations were observed for fasting glucose, HDL-C, CRP, or BMI. Conclusions: Vitamin D deficiency was independently associated with poorer long-term glycemic status, hypertriglyceridemia, and central adiposity in older adults, but not with other metabolic markers after adjustment. These findings suggest that the metabolic correlates of vitamin D deficiency may be domain-specific rather than generalized. Longitudinal and interventional studies are needed to clarify causality and underlying mechanisms. Full article

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

14 pages, 680 KB

Open AccessEditor’s ChoiceReview

The Thyroid–Metabolism Axis: Pathways of Dysregulation and the Effects of Treatment

by Martina Curcio and Royce P. Vincent

Metabolites 2026, 16(4), 267; https://doi.org/10.3390/metabo16040267 - 16 Apr 2026

Viewed by 1334

Abstract

Thyroid hormones regulate a complex and interconnected network of metabolic signaling. Thyroid dysfunction is, at present, defined and monitored through circulating thyroid-stimulating hormone (TSH) and free thyroid hormones. However, biochemical normalization does not entirely indicate restoration of metabolic homeostasis. This discrepancy highlights a [...] Read more.

Thyroid hormones regulate a complex and interconnected network of metabolic signaling. Thyroid dysfunction is, at present, defined and monitored through circulating thyroid-stimulating hormone (TSH) and free thyroid hormones. However, biochemical normalization does not entirely indicate restoration of metabolic homeostasis. This discrepancy highlights a critical limitation of the current TSH-centric paradigm, which also fails to explain the heterogeneity in cardiometabolic outcomes observed among patients with similar biochemical profiles. Metabolomics, through the analysis of tissue-specific biofluids, could aid in capturing the complex metabolic perturbations that characterize this disease. In this review, we summarize metabolomic signatures typical of thyroid dysfunction, perform a critical evaluation of limitations and variability across studies, and explore the clinical and translational implications of metabolomics in thyroid pathology. In addition, five metabolic hubs influenced by thyroid hormone activity are summarized: (i) lipid and lipoprotein remodeling; (ii) mitochondrial energetics and redox balance; (iii) amino acid metabolism and protein turnover; (iv) gut–liver–thyroid axis and (v) biological impact of subclinical thyroid diseases. Taken together, these findings challenge the sufficiency of a diagnostic model based on TSH measurement and pose metabolomics as a promising tool to refine risk stratification, uncover subclinical vulnerability and guide patient-centered management of thyroid disease. Despite its promise, clinical adoption of metabolomics is hindered by a lack of standardization and complex data interpretation. To overcome these limitations, coupling metabolomics with genomics and transcriptomics may allow its translation into practical application. Full article

(This article belongs to the Special Issue Exploring Metabolomic Signatures and the Metabolic Impact of Thyroid Dysfunction)

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18 pages, 1044 KB

Open AccessEditor’s ChoiceArticle

Effects of Probiotic Supplementation on Gut Microbiota and Fecal Metabolome in Autism Spectrum Disorders: A Secondary Analysis of a Randomized Clinical Trial in Preschoolers

by Letizia Guiducci, Luca Laghi, Nicolò Dellarosa, Paola Mastromarino, Margherita Prosperi, Filippo Muratori and Sara Calderoni

Metabolites 2026, 16(4), 262; https://doi.org/10.3390/metabo16040262 - 13 Apr 2026

Viewed by 850

Abstract

Background/Objectives: Recently, a randomized clinical trial evaluated whether a six-month probiotic administration could reduce symptom severity in preschool children with Autism Spectrum Disorders (ASD), with (GI) or without (NGI) gastrointestinal symptoms. Significant positive changes were observed only in NGI children. A second explorative [...] Read more.

Background/Objectives: Recently, a randomized clinical trial evaluated whether a six-month probiotic administration could reduce symptom severity in preschool children with Autism Spectrum Disorders (ASD), with (GI) or without (NGI) gastrointestinal symptoms. Significant positive changes were observed only in NGI children. A second explorative study on children prior to intervention identified a fecal metabolome fingerprint associated with ASD severity. Building on these findings, the present study aimed to assess whether metabolomics could monitor changes in ASD severity following probiotic administration using a subset of samples from the same trial. Second, this study aimed to identify fecal metabolites to be monitored in children to predict whether their autism severity may decrease after probiotic or placebo treatment. Methods: Evaluations of the fecal metabolome and microbiota could be completed on 57 children before and after a double-blind administration of a probiotic mixture or a placebo. Results: In NGI children the probiotic was found to influence the concentration of the amino acids aspartate, leucine, tryptophan, and valine, together with nicotinate and the short chain fatty acids acetate, butyrate, isobutyrate, and propionate. Lactobacilli and Sutterella showed significant changes in response to probiotic administration (p < 0.05). Acetate, 4-hydroxyphenyl, galactose, proline, and tyramine were identified as key fecal metabolites for prediction purposes. Conclusions: The present exploratory analysis, despite the small sample size, suggests that fecal metabolomics may provide a useful approach for monitoring and potentially for predicting changes in ASD severity following probiotics administration. Full article

(This article belongs to the Special Issue Advances in Metabolomics for Precision Medicine: From Biomarker Discovery to Clinical Applications: 2nd Edition)

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19 pages, 915 KB

Open AccessEditor’s ChoiceReview

A Dual-Target Microbial Therapeutic Strategy for Treating Metabolic Diseases: Complementary Mechanisms and Clinical Prospects of Lactiplantibacillus plantarum and Akkermansia muciniphila

by Si Liu, Mao Wang, Xiaobo Sun, Zhihao Jia and Kuilong Huang

Metabolites 2026, 16(4), 259; https://doi.org/10.3390/metabo16040259 - 13 Apr 2026

Cited by 1 | Viewed by 1203

Abstract

Metabolic diseases, including obesity, type 2 diabetes, and their related complications, have emerged as major global public health challenges. Increasing evidence indicates that gut microbiota dysbiosis contributes to disrupted metabolic homeostasis, chronic low-grade inflammation, and progression of metabolic disorders. Among candidate microbiome-based interventions, [...] Read more.

Metabolic diseases, including obesity, type 2 diabetes, and their related complications, have emerged as major global public health challenges. Increasing evidence indicates that gut microbiota dysbiosis contributes to disrupted metabolic homeostasis, chronic low-grade inflammation, and progression of metabolic disorders. Among candidate microbiome-based interventions, Lactiplantibacillus plantarum (L. plantarum) and Akkermansia muciniphila (A. muciniphila) have attracted particular attention because they regulate host metabolism through partially distinct yet potentially complementary mechanisms. L. plantarum has been associated with modulation of appetite-related hormones, adipose tissue remodeling, reinforcement of intestinal barrier function, and attenuation of inflammatory signaling. A. muciniphila has been linked to strengthening of the mucus barrier, production of beneficial metabolites, and improvement in immune and metabolic homeostasis. However, current evidence remains fragmented across strain-specific studies, heterogeneous formulations, and predominantly single-strain experimental designs, and direct comparative evidence for combined administration is still limited. This review synthesizes current epidemiological, mechanistic, preclinical, and clinical evidence on L. plantarum and A. muciniphila, with emphasis on their physiological traits, gut ecological adaptability, pathway-based metabolic effects, and translational challenges in obesity, type 2 diabetes, and related complications. We further highlight the ecological rationale for their functional complementarity and discuss priorities for future combination studies and precision implementation. Overall, the available literature supports functional complementarity and possible additive metabolic benefits, but synergistic effects in humans remain unconfirmed. A clearer understanding of strain identity, active therapeutic entities, delivery strategies, and host context will be essential for advancing this dual-target microbial strategy toward clinically meaningful applications. Full article

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

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18 pages, 3377 KB

Open AccessEditor’s ChoiceArticle

Age-Specific Ex Vivo Modulation of Gut–Brain Axis-Associated Metabolites by Galacto-Oligosaccharides and Nutrient Blends in Early Childhood

by Laurent Ferrier, Shaillay Kumar Dogra, Lam Dai Vu, Alexandros K. Kanellopoulos, Jonas Poppe, Laurence Biehl, Aurélien Baudot and Pieter Van den Abbeele

Metabolites 2026, 16(4), 255; https://doi.org/10.3390/metabo16040255 - 10 Apr 2026

Viewed by 1424

Abstract

Background: Gut microbiome-derived metabolites, particularly short-chain fatty acids (SCFA) and tryptophan derivatives, are central mediators of the gut–brain axis. This ex vivo study assessed how nutritional interventions impact such metabolites during early life, a critical period for neurodevelopment. Methods: The effects [...] Read more.

Background: Gut microbiome-derived metabolites, particularly short-chain fatty acids (SCFA) and tryptophan derivatives, are central mediators of the gut–brain axis. This ex vivo study assessed how nutritional interventions impact such metabolites during early life, a critical period for neurodevelopment. Methods: The effects of galacto-oligosaccharides (GOS), nutrient blends (vitamins, minerals and amino acids) and their combinations were evaluated in the gut microbiomes of infants (2–4 months, n = 6) and young children (2–3 years old, n = 6) using the ex vivo SIFR^® technology. Results: Baseline microbiome composition was age-dependent, with infants displaying lower α-diversity and greater interpersonal variability. After ex vivo incubation, nutrient blends increased the propionate/butyrate ratio and branched-chain fatty acids in young children and elevated several B-vitamins and amino acid-derived metabolites, including indole-3-carboxaldehyde, imidazoleacetic acid and pipecolinic acid. Combining nutrient blends with GOS exhibited potential synergistic effects on propionate (infants) and 2-hydroxyisocaproic acid (HICA, both age groups). GOS strongly stimulated Bifidobacteriaceae and increased metabolites linked to bifidobacterial metabolism like acetate, HICA, N-acetylated amino acids, aromatic lactic acids and acetylagmatine; in young children, butyrate and γ-aminobutyric acid (GABA) also increased. Conclusions: Combinations of GOS with nutrient blends impacted microbiome-derived metabolites associated with the gut–brain axis, with potential synergistic increases of metabolites with emerging roles in neurodevelopment, including GABA, acetylagmatine and HICA. Despite shared bifidogenic effects, differences between age groups indicate that microbiome maturity may influence responses to nutritional intervention. Future clinical studies are needed to determine whether these metabolite changes translate into neurodevelopmental benefits in vivo. Full article

(This article belongs to the Special Issue Neuronutrition: Metabolomic Insights and Perspectives)

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15 pages, 281 KB

Open AccessEditor’s ChoiceArticle

Evaluation of Nutritional, Antioxidant, Antidiabetic, and Antidyslipidemic Properties of Red Corn Tortillas Enriched with Moringa oleifera Leaves

by Eunice Tranquilino-Rodríguez, Noé Calderón-Téllez, José Juan Virgen-Ortiz, Juan de Dios Figueroa-Cárdenas, Rafael Zamora-Vega, José Octavio Rodiles-López and Héctor Eduardo Martínez-Flores

Metabolites 2026, 16(4), 252; https://doi.org/10.3390/metabo16040252 - 8 Apr 2026

Viewed by 882

Abstract

Background/Objectives: Metabolic diseases are increasingly associated with diets low in bioactive compounds. Native maize varieties possess functional potential; however, they remain underutilized. Moringa oleifera leaf flour (MF), rich in protein and polyphenols, represents a promising functional ingredient. This study evaluated the incorporation of [...] Read more.

Background/Objectives: Metabolic diseases are increasingly associated with diets low in bioactive compounds. Native maize varieties possess functional potential; however, they remain underutilized. Moringa oleifera leaf flour (MF), rich in protein and polyphenols, represents a promising functional ingredient. This study evaluated the incorporation of MF into red native corn tortillas and its effects on nutritional composition and antioxidant capacity, as well as assessed its hypoglycemic and hypolipidemic effects in Wistar rats. Methods: Tortillas were formulated with 5% MF. Nutritional composition was determined using standard AOAC methods, while bioactive compounds (total phenolics and flavonoids) and antioxidant activity were evaluated using Folin–Ciocalteu, aluminum chloride (AlCl₃) colorimetric, DPPH^•, and ABTS^•+ assays, respectively. Male Wistar rats (12 weeks old, with an approximate weight ofs 360 g; n = 5/group) were fed the experimental diets for 21 days with either a standard diet, a high-fat diet, or high-fat diets supplemented with MF or MF-enriched tortillas. Serum glucose, triglycerides, total cholesterol, and HDL were measured using enzymatic colorimetric methods. Data were analyzed by ANOVA followed by Tukey’s test (p < 0.05). Results: MF incorporation increased protein (+19.85%), dietary fiber (+18.51%), and mineral content (+41.03%) compared to control tortillas. Total phenolics and flavonoids increased by 114.0% and 184.7%, respectively. Antioxidant activity improved significantly, as evidenced by reductions in IC₅₀ values of 41.1% (DPPH^•) and 43.1% (ABTS^•). In vivo, MF-enriched tortillas reduced triglycerides by 68.4%, total cholesterol by 16.2%, and hepatic lipid accumulation by 31.8% compared to the high-fat diet group. Glucose levels showed a reduction of 8.5%, although not statistically significant (p > 0.05). Conclusions: The incorporation of MF into red corn tortillas significantly enhances their nutritional and functional properties. In vivo results also showed improvements in lipid profile and a non-significant reduction in glucose levels. These findings support the development of functional foods based on traditional staples with potential health benefits. Full article

(This article belongs to the Section Nutrition and Metabolism)

12 pages, 219 KB

Open AccessEditor’s ChoiceArticle

Personalizing Obesity Treatment: Real-World Comparison of a Very-Low-Calorie Ketogenic Diet Versus a Whole-Food Mediterranean Ketogenic Diet

by Davide Masi, Maria Letizia Spizzichini, Elena Colonnello, Daniel Vasquez Barahona, Lucio Gnessi, Daniele Gianfrilli and Mikiko Watanabe

Metabolites 2026, 16(4), 248; https://doi.org/10.3390/metabo16040248 - 5 Apr 2026

Cited by 1 | Viewed by 1724

Abstract

Background/Objectives: Obesity is a chronic, relapsing disease in which lifestyle modification represents the cornerstone of treatment. Among dietary strategies, ketogenic diets can induce rapid weight loss, whereas the Mediterranean diet is associated with established cardiometabolic benefits but typically produces slower weight reduction. Very-low-calorie [...] Read more.

Background/Objectives: Obesity is a chronic, relapsing disease in which lifestyle modification represents the cornerstone of treatment. Among dietary strategies, ketogenic diets can induce rapid weight loss, whereas the Mediterranean diet is associated with established cardiometabolic benefits but typically produces slower weight reduction. Very-low-calorie ketogenic diets (VLCKDs) are effective for weight loss but are often limited by cost, reliance on meal replacements, and reduced long-term feasibility. This study aimed to evaluate whether a whole-food Mediterranean ketogenic diet with moderate caloric restriction (MedKD) could represent a feasible and effective alternative to VLCKD for weight loss and metabolic improvement in adults with obesity. Methods: This 3-month prospective, real-world study compared VLCKD and MedKD in adults with obesity attending a clinical nutrition program. The primary outcome was percentage weight loss. Secondary outcomes included changes in waist circumference, waist-to-height ratio, insulin resistance (HOMA-IR), lipid profile, kidney function, and treatment tolerability. Clinical and biochemical parameters were assessed at baseline and after the intervention. Group differences and time-by-group interactions were analyzed to evaluate changes over the study period. Results: Sixty-two participants were enrolled, and 55 completed the study (27 VLCKD, 28 MedKD). Baseline characteristics were generally comparable, although the MedKD group had a higher prevalence of diabetes and higher baseline insulin resistance and triglyceride levels. Both dietary interventions resulted in substantial and comparable weight loss (approximately 15% of initial body weight), accompanied by significant reductions in waist circumference and waist-to-height ratio. Insulin resistance improved in both groups, with a greater reduction in HOMA-IR observed in the MedKD group (time × group p = 0.031). Serum creatinine decreased in the VLCKD group and slightly increased in the MedKD group (p = 0.025). Changes in lipid profile were not significantly different between groups. No severe adverse events were reported. Conclusions: A whole-food Mediterranean ketogenic diet with moderate caloric restriction achieved weight loss and metabolic improvements comparable to those observed with VLCKD over three months. These findings suggest that MedKD may represent a feasible alternative to formula-based ketogenic programs, supporting more flexible and personalized dietary strategies in the clinical management of obesity. Full article

(This article belongs to the Special Issue Diabetes and Metabolic Diseases: From Prevention to Clinical Management, 2nd Edition)

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