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Fibromyalgia (FM), a chronic disease with a high incidence in women, poses a significant challenge for diagnosis and treatment, especially due to the absence of specific biomarkers and the multifaceted nature of its symptoms, which range from neuromuscular pain to mood disorders and intestinal dysbiosis. While diagnosis currently relies on rheumatological clinical evaluations and treatment options mainly focus on symptom management, FM seems to have possible links with systemic metabolic dysfunctions with a common inflammatory root. In this context, a new therapeutic avenue emerges: could a therapeutic nutritional approach be the missing piece of the puzzle? Indeed, diet therapies employed particularly for metabolic syndromes proved recently to be efficacious for correcting systemic dysmetabolism and a high number of chronic inflammation conditions. In particular, the very-low-calorie ketogenic diet (VLCKD) demonstrated therapeutic benefits in many disorders. In the present study, we aimed to investigate the specific effects of two dietary interventions, namely the oloproteic VLCKD and the low-glycemic insulinemic (LOGI) diet, on two groups of female FM patients (FM1 and FM2) over a 45-day period. Utilizing clinical and laboratory tests, as well as non-invasive NMR metabolomic analysis of serum, urine, and saliva samples, we sought to uncover how these dietary regimens impact the metabolic dysfunctions associated with FM. [ABSTRACT FROM AUTHOR]

Introduction: Due to its peculiar anatomy and physiology, the pericardial fluid is a biological matrix of particular interest in the forensic field. Despite this, the available literature has mainly focused on post-mortem biochemistry and forensic toxicology, while to the best of authors' knowledge post-mortem metabolomics has never been applied. Similarly, estimation of the time since death or post-mortem interval based on pericardial fluids has still rarely been attempted. Objectives: We applied a metabolomic approach based on 1H nuclear magnetic resonance spectroscopy to ascertain the feasibility of monitoring post-mortem metabolite changes on human pericardial fluids with the aim of building a multivariate regression model for post-mortem interval estimation. Methods: Pericardial fluid samples were collected in 24 consecutive judicial autopsies, in a time frame ranging from 16 to 170 h after death. The only exclusion criterion was the quantitative and/or qualitative alteration of the sample. Two different extraction protocols were applied for low molecular weight metabolites selection, namely ultrafiltration and liquid-liquid extraction. Our metabolomic approach was based on the use of 1H nuclear magnetic resonance and multivariate statistical data analysis. Results: The pericardial fluid samples treated with the two experimental protocols did not show significant differences in the distribution of the metabolites detected. A post-mortem interval estimation model based on 18 pericardial fluid samples was validated with an independent set of 6 samples, giving a prediction error of 33–34 h depending on the experimental protocol used. By narrowing the window to post-mortem intervals below 100 h, the prediction power of the model was significantly improved with an error of 13–15 h depending on the extraction protocol. Choline, glycine, ethanolamine, and hypoxanthine were the most relevant metabolites in the prediction model. Conclusion: The present study, although preliminary, shows that PF samples collected from a real forensic scenario represent a biofluid of interest for post-mortem metabolomics, with particular regard to the estimation of the time since death. [ABSTRACT FROM AUTHOR]

Search for natural sources in disease management especially diabetes has surged recently for its potential health benefits. However, several fruit tree species remained unpopular despite their folkloric usage due to insufficient scientific data. Syzygium jambos (L.) Alston (Myrtaceae) is a fruit-bearing plant that is used as a traditional dietary supplement to treat various diseases including diabetes. This study investigated the correlation between S. jambos leaves’ metabolites and the bioactivities using proton-nuclear magnetic resonance (1H NMR)-based metabolomics approach. In-vitro total phenolic content (TPC), 2,2-diphenyl-1-picrylhydrazyl (DPPH•) radicals scavenging, nitric oxide (NO•) radicals scavenging, anti-α-amylase and anti-α-glucosidase assays were performed. The findings indicated that 70 % ethanolic extract demonstrated the highest potential in overall bioactivities. A total of 59 and 30 metabolites were tentatively identified using ultrahigh-performance liquid chromatography electrospray ionization Quadrupole-Orbitrap tandem mass spectrometry (UHPLC/ESI Q-Orbitrap MS/MS) and NMR, respectively. The partial least square (PLS) model revealed tannins, triterpenoids, and flavonoids significantly contributed to the separation and bioactivities. This study provides comprehensive insights into S. jambos metabolome and reveals the potential as a reliable source of antioxidant and anti-hyperglycemic compounds.

Fibromyalgia (FM), a chronic disease with a high incidence in women, poses a significant challenge for diagnosis and treatment, especially due to the absence of specific biomarkers and the multifaceted nature of its symptoms, which range from neuromuscular pain to mood disorders and intestinal dysbiosis. While diagnosis currently relies on rheumatological clinical evaluations and treatment options mainly focus on symptom management, FM seems to have possible links with systemic metabolic dysfunctions with a common inflammatory root. In this context, a new therapeutic avenue emerges: could a therapeutic nutritional approach be the missing piece of the puzzle? Indeed, diet therapies employed particularly for metabolic syndromes proved recently to be efficacious for correcting systemic dysmetabolism and a high number of chronic inflammation conditions. In particular, the very-low-calorie ketogenic diet (VLCKD) demonstrated therapeutic benefits in many disorders. In the present study, we aimed to investigate the specific effects of two dietary interventions, namely the oloproteic VLCKD and the low-glycemic insulinemic (LOGI) diet, on two groups of female FM patients (FM1 and FM2) over a 45-day period. Utilizing clinical and laboratory tests, as well as non-invasive NMR metabolomic analysis of serum, urine, and saliva samples, we sought to uncover how these dietary regimens impact the metabolic dysfunctions associated with FM.

Peony root is an important herbal drug used as an antispasmodic analgesic. To evaluate peony roots with different botanical origins, producing areas, and post-harvest processing, 1H NMR-based metabolomics analysis was employed. Five types of monoterpenoids, including albiflorin (4), paeoniflorin (6), and sulfonated paeoniflorin (25), and six other compounds, including 1,2,3,4,6-penta-O-galloyl-β-d-glucose (18), benzoic acid (21), gallic acid (22), and sucrose (26) were detected in the extracts of peony root samples. Among them, compounds 4, 6, 18, and total monoterpenoids including 21 were quantified by quantitative 1H NMR (qHNMR). Compound 25 was detected in 1H NMR spectra of sulfur-fumigated white peony root (WPR) extracts indicating that 1H NMR was a fast and effective method for identifying sulfur-fumigated WPR. The content of 26, the main factor affecting extract yield, increased significantly in peony root after low-temperature storage for one month, whereas that in WPR did not increase due to the boiling treatment after harvesting. We investigated the impact of preprocessing methods to such analysis for NMR data from commercial samples, resulting that the data matrix transformed from qHNMR spectra and normalized to internal standard were optimum for multivariate analysis. The multivariate analysis demonstrated that among commercial samples derived from P. lactiflora, peony root samples in Japanese market (PR) had high contents of 18 and 22, and red peony root (RPR) samples had high content of monoterpenoids represented by 6; and among RPR samples, those derived from P. veitchii showed higher contents of 18 and 22 than those from P. lactiflora. The 1H NMR-based metabolomics method coupled with qHNMR was useful for evaluation of peony root and would be applicable for other crude drugs. [ABSTRACT FROM AUTHOR]

Annona muricata is a common plant used in Africa and South America to manage various types of disease. However, there is insufficient toxicological information or published standard available regarding repeated dose animal toxicity data. As part of the safety assessment, we exposed Sprague Dawley rats to an acute oral toxicity of A. muricata. The intent of the current study was to use advanced proton nuclear magnetic resonance (1H NMR) in serum and urinary metabolomics evaluation techniques to provide the in vivo acute toxicological profile of A. muricata leaf ethanol extract in accordance with the Organization for Economic Co-operation and Development's (OECD) 423 guidelines. A single 2000 mg/kg dose of A. muricata leaf ethanol extract was administered to Sprague Dawley rats over an observational period of 14 days. The toxicity evaluation (physical and behavior observation, body weight, renal function test, liver function test and 1H NMR analysis) showed no abnormal toxicity. Histopathological analysis manifested mild changes, i.e., the treated kidney manifested mild hypercellularity of mesangial cells and mild red blood cell congestion. In addition, there was mild hemorrhage into tissue with scattered inflammatory cells and mild dilated central vein with fibrosis in the liver. However, the changes were very mild and not significant which correlate with other analyses conducted in this study (biochemical test and 1H NMR metabolomic analysis). On the other hand, urinary 1H NMR analysis collected on day 15 revealed high similarity on the metabolite variations for both untreated and treated groups. Importantly, the outcomes suggest that A. muricata leaf ethanol extract can be safely consumed at a dose of 2000 mg/kg and the LD50 must be more than 2000 mg/kg. [ABSTRACT FROM AUTHOR]

Introduction: The estimation of post-mortem interval (PMI) remains a major challenge in forensic science. Most of the proposed approaches lack the reliability required to meet the rigorous forensic standards. Objectives: We applied 1H NMR metabolomics to estimate PMI on ovine vitreous humour comparing the results with the actual scientific gold standard, namely vitreous potassium concentrations. Methods: Vitreous humour samples were collected in a time frame ranging from 6 to 86 h after death. Experiments were performed by using 1H NMR metabolomics and ion capillary analysis. Data were submitted to multivariate statistical data analysis. Results: A multivariate calibration model was built to estimate PMI based on 47 vitreous humour samples. The model was validated with an independent test set of 24 samples, obtaining a prediction error on the entire range of 6.9 h for PMI < 24 h, 7.4 h for PMI between 24 and 48 h, and 10.3 h for PMI > 48 h. Time-related modifications of the 1H NMR vitreous metabolomic profile could predict PMI better than potassium up to 48 h after death, whilst a combination of the two is better than the single approach for higher PMI estimation. Conclusion: The present study, although in a proof-of-concept animal model, shows that vitreous metabolomics can be a powerful tool to predict PMI providing a more accurate estimation compared to the widely studied approach based on vitreous potassium concentrations. [ABSTRACT FROM AUTHOR]

In the absence of appearance features, it is becoming increasingly difficult to accurately distinguish botanical herbal powders from different families or parts of the plant. In this study, we explored the biomarkers with neuroprotective effects in multi-parts ginseng based on metabolite content changes and biological functions. First, using 1H NMR and LC-MS metabolomics techniques and multivariate statistical analysis techniques to analyze the differential metabolites (DMs) in ginseng main root (TR), lateral root (LR) and Rhizome (RZ). Also, the analysis of 14 differential metabolites (DMs) using network pharmacology yielded 23 core targets associated with neuroprotective diseases, the KEGG pathway was enriched for 117 signaling pathways and 184 enrichment processes were shown in the GO analysis. Combined with the correlation analysis of primary differential metabolites (PDMs) and secondary differential metabolites (SDMs) content and network activity, we finally identified three biomarkers in different parts of ginseng, namely sucrose, ginsenoside Rb1, and ginsenoside Rb3. Finally, the three biomarkers were validated by molecular docking and SH-SY5Y survival, and the results showed that sucrose, ginsenoside Rb1 and ginsenoside Rb3 exhibited favorable activity results in neuroprotective functions. The results of this study reveal the overall differences in the DMs of the three parts of ginseng, and three biomarkers can help identify different parts of ginseng powder, to avoid illegal substitution when ginseng is used in medicine. At the same time, it also provides a reference method for screening biomarkers that can distinguish different parts of traditional Chinese medicine.

Annona muricata is a common plant used in Africa and South America to manage various types of disease. However, there is insufficient toxicological information or published standard available regarding repeated dose animal toxicity data. As part of the safety assessment, we exposed Sprague Dawley rats to an acute oral toxicity of A. muricata. The intent of the current study was to use advanced proton nuclear magnetic resonance (1H NMR) in serum and urinary metabolomics evaluation techniques to provide the in vivo acute toxicological profile of A. muricata leaf ethanol extract in accordance with the Organization for Economic Co-operation and Development’s (OECD) 423 guidelines. A single 2000 mg/kg dose of A. muricata leaf ethanol extract was administered to Sprague Dawley rats over an observational period of 14 days. The toxicity evaluation (physical and behavior observation, body weight, renal function test, liver function test and 1H NMR analysis) showed no abnormal toxicity. Histopathological analysis manifested mild changes, i.e., the treated kidney manifested mild hypercellularity of mesangial cells and mild red blood cell congestion. In addition, there was mild hemorrhage into tissue with scattered inflammatory cells and mild dilated central vein with fibrosis in the liver. However, the changes were very mild and not significant which correlate with other analyses conducted in this study (biochemical test and 1H NMR metabolomic analysis). On the other hand, urinary 1H NMR analysis collected on day 15 revealed high similarity on the metabolite variations for both untreated and treated groups. Importantly, the outcomes suggest that A. muricata leaf ethanol extract can be safely consumed at a dose of 2000 mg/kg and the LD50 must be more than 2000 mg/kg.

The integration of cell metabolism with signalling pathways, transcription factor networks and epigenetic mediators is critical in coordinating molecular and cellular events during embryogenesis. Induced pluripotent stem cells (IPSCs) are an established model for embryogenesis, germ layer specification and cell lineage differentiation, advancing the study of human embryonic development and the translation of innovations in drug discovery, disease modelling and cell-based therapies. The metabolic regulation of IPSC pluripotency is mediated by balancing glycolysis and oxidative phosphorylation, but there is a paucity of data regarding the influence of individual metabolite changes during cell lineage differentiation. We used 1H NMR metabolite fingerprinting and footprinting to monitor metabolite levels as IPSCs are directed in a three-stage protocol through primitive streak/mesendoderm, mesoderm and chondrogenic populations. Metabolite changes were associated with central metabolism, with aerobic glycolysis predominant in IPSC, elevated oxidative phosphorylation during differentiation and fatty acid oxidation and ketone body use in chondrogenic cells. Metabolites were also implicated in the epigenetic regulation of pluripotency, cell signalling and biosynthetic pathways. Our results show that 1H NMR metabolomics is an effective tool for monitoring metabolite changes during the differentiation of pluripotent cells with implications on optimising media and environmental parameters for the study of embryogenesis and translational applications. [ABSTRACT FROM AUTHOR]

The Aeolian Islands (Italy) are a volcanic archipelago in the Tyrrhenian Sea comprising seven main islands, among which are two active volcanoes. The peculiar geological features and the wide variety of environments and soils have an important impact on native plants, and in particular, the Aeolian populations of Dactylis glomerata (a perennial cool-season bunchgrass) exhibit remarkable phenotypic variability. Considering that environmental drivers also strongly affect the production of plant metabolites, this work aimed at comparing the metabolomic profiles of D. glomerata (leaves) harvested at different altitudes on four islands of the Aeolian archipelago, namely: Lipari, Vulcano, Stromboli and Panarea. Samples were analyzed by 1H NMR profiling, and data were treated by PCA. Samples collected on Stromboli were very different from each other and from the samples collected in the other islands. Through an Orthogonal Partial Least Squares (OPLS) model, using altitude as the y variable, it emerged that the concentration of proline, glycine betaine, sucrose, glucose and chlorogenic acid of D. glomerata growing on Stromboli decreased at increasing altitude. Conversely, increasing altitude was associated with an increment in valine, asparagine, fumaric acid and phenylalanine. [ABSTRACT FROM AUTHOR]

This study evaluated the cytotoxic activity of Tamoxifen (TMX), an anti-estrogen drug, with microalgal crude extracts (MCEs) in single and synergistic application (TMX-MCEs) on MCF-7 and 4T1 breast cancer cells, and non-cancerous Vero cells. The MCEs of Nannochloropsis oculata, Tetraselmis suecica and Chlorella sp. from five different solvents (methanol, MET; ethanol, ETH; water, W; chloroform, CHL; and hexane, HEX) were developed. The TMX-MCEs-ETH and W at the 1:2 and 1:3 ratios, attained IC50 of 15.84–29.51 μg/mL against MCF-7; 13.8–31.62 μg/mL against 4T1; and 24.54–85.11 μg/mL against Vero cells. Higher late apoptosis was exhibited against MCF-7 by the TMX-N. oculata-ETH (41.15 %); and by the TMX-T. suecica-ETH (65.69 %) against 4T1 cells. The TMX-T. suecica-ETH also showed higher ADP/ATP ratios, but comparable Caspase activities to control. For Vero cells, overall apoptotic effects were lowered with synergistic application, and only early apoptosis was higher with TMX-T. suecica-ETH but at lower levels (29.84 %). The MCEs-W showed the presence of alanine, oleic acid, linoleic acid, lactic acid, and fumaric acid. Based on Principal Component Analysis (PCA), the spectral signals for polar solvents such as MET and ETH, were found in the same cluster, while the non-polar solvent CHL was with HEX, suggesting similar chemical profiles clustered for the same polarity. The CHL and HEX were more effective with N. oculata and T. suecica which were of the marine origin, while the ETH and MET were more effective with Chlorella sp., which was of the freshwater origin. The synergistic application of microalgal bioactive compounds with TMX can maintain the cytotoxicity against breast cancer cells whilst reducing the toxicity against non-cancerous Vero cells. These findings will benefit the biopharmaceutical, and functional and healthy food industries.

The integration of cell metabolism with signalling pathways, transcription factor networks and epigenetic mediators is critical in coordinating molecular and cellular events during embryogenesis. Induced pluripotent stem cells (IPSCs) are an established model for embryogenesis, germ layer specification and cell lineage differentiation, advancing the study of human embryonic development and the translation of innovations in drug discovery, disease modelling and cell-based therapies. The metabolic regulation of IPSC pluripotency is mediated by balancing glycolysis and oxidative phosphorylation, but there is a paucity of data regarding the influence of individual metabolite changes during cell lineage differentiation. We used 1H NMR metabolite fingerprinting and footprinting to monitor metabolite levels as IPSCs are directed in a three-stage protocol through primitive streak/mesendoderm, mesoderm and chondrogenic populations. Metabolite changes were associated with central metabolism, with aerobic glycolysis predominant in IPSC, elevated oxidative phosphorylation during differentiation and fatty acid oxidation and ketone body use in chondrogenic cells. Metabolites were also implicated in the epigenetic regulation of pluripotency, cell signalling and biosynthetic pathways. Our results show that 1H NMR metabolomics is an effective tool for monitoring metabolite changes during the differentiation of pluripotent cells with implications on optimising media and environmental parameters for the study of embryogenesis and translational applications.

Estimation of the post-mortem interval (PMI) remains a matter of concern in the forensic scenario. Traditional and novel approaches are not yet able to fully address this issue, which relies on complex biological phenomena triggered by death. For this purpose, eye compartments may be chosen for experimental studies because they are more resistant to post-mortem modifications. Vitreous humour, in particular, has been extensively investigated, with potassium concentration ([K+]) being the marker that is better correlated with PMI estimation. Recently, a 1H nuclear magnetic resonance (NMR) metabolomic approach based on aqueous humour (AH) from an animal model was proposed for PMI estimation, resulting in a robust and validated regression model. Here we studied the variation in [K+] in the same experimental setup. [K+] was determined through capillary ion analysis (CIA) and a regression analysis was performed. Moreover, it was investigated whether the PMI information related to potassium could improve the metabolome predictive power in estimating the PMI. Interestingly, we found that a part of the metabolomic profile is able to explain most of the information carried by potassium, suggesting that the rise in both potassium and metabolite concentrations relies on a similar biological mechanism. In the first 24-h PMI window, the AH metabolomic profile shows greater predictive power than [K+] behaviour, suggesting its potential use as an additional tool for estimating the time since death. [ABSTRACT FROM AUTHOR]

• NMR and UHPLC-MS/MS identified 30 and 59 metabolites respectively in S. jambos leaf. • S. jambos leaf extract showed antioxidant and anti-hyperglycemic properties. • 70% ethanolic extract is the most active in all bioactivities tested. • Tannins, triterpenoids and flavonoids contributed to the bioactivities. • NMR metabolomics reveals the potential of S. jambos as functional ingredients. Search for natural sources in disease management especially diabetes has surged recently for its potential health benefits. However, several fruit tree species remained unpopular despite their folkloric usage due to insufficient scientific data. Syzygium jambos (L.) Alston (Myrtaceae) is a fruit-bearing plant that is used as a traditional dietary supplement to treat various diseases including diabetes. This study investigated the correlation between S. jambos leaves' metabolites and the bioactivities using proton-nuclear magnetic resonance (1H NMR)-based metabolomics approach. In-vitro total phenolic content (TPC), 2,2-diphenyl-1-picrylhydrazyl (DPPH•) radicals scavenging, nitric oxide (NO•) radicals scavenging, anti-α-amylase and anti-α-glucosidase assays were performed. The findings indicated that 70 % ethanolic extract demonstrated the highest potential in overall bioactivities. A total of 59 and 30 metabolites were tentatively identified using ultrahigh-performance liquid chromatography electrospray ionization Quadrupole-Orbitrap tandem mass spectrometry (UHPLC/ESI Q-Orbitrap MS/MS) and NMR, respectively. The partial least square (PLS) model revealed tannins, triterpenoids, and flavonoids significantly contributed to the separation and bioactivities. This study provides comprehensive insights into S. jambos metabolome and reveals the potential as a reliable source of antioxidant and anti-hyperglycemic compounds. [ABSTRACT FROM AUTHOR]

The Aeolian Islands (Italy) are a volcanic archipelago in the Tyrrhenian Sea comprising seven main islands, among which are two active volcanoes. The peculiar geological features and the wide variety of environments and soils have an important impact on native plants, and in particular, the Aeolian populations of Dactylis glomerata (a perennial cool-season bunchgrass) exhibit remarkable phenotypic variability. Considering that environmental drivers also strongly affect the production of plant metabolites, this work aimed at comparing the metabolomic profiles of D. glomerata (leaves) harvested at different altitudes on four islands of the Aeolian archipelago, namely: Lipari, Vulcano, Stromboli and Panarea. Samples were analyzed by 1H NMR profiling, and data were treated by PCA. Samples collected on Stromboli were very different from each other and from the samples collected in the other islands. Through an Orthogonal Partial Least Squares (OPLS) model, using altitude as the y variable, it emerged that the concentration of proline, glycine betaine, sucrose, glucose and chlorogenic acid of D. glomerata growing on Stromboli decreased at increasing altitude. Conversely, increasing altitude was associated with an increment in valine, asparagine, fumaric acid and phenylalanine.

Over the past decade, 1H NMR metabolomic studies have become increasingly popular in wine research since they can provide a chemical fingerprint with information regarding authenticity, geographical origin, vintage, and wine composition. Comparatively few 1H NMR-based studies have evaluated sparkling wines. In the present study, quantitative 1H NMR metabolomics was applied to assess the composition of finished traditional method (bottle-fermented) sparkling wines aged for 18-months in cellar conditions (14 °C, 70% relative humidity) following liqueur d′expedition (dosage) additions with six sugar types (glucose, fructose, sucrose derived from sugar beets and sugar cane, maltose, and MCR Sucraisin®) and a zero- dosage control. No systematic trends were observed when comparing the metabolite profile across sugar types, nor between the zero- dosage control and treatments with sugar additions. Multivariate statistical approaches using principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA) did not effectively discriminate between dosage treatments nor wine age, while orthogonal projections to latent structures discriminant analysis (OPLS-DA) successfully discriminated between wines by age and was validated through permutation tests. This study presents the first application of 1H NMR metabolomics coupled with multivariate analysis to monitor the aging of sparkling wines. [Display omitted] • Novel application of 1H NMR metabolomics to aged sparkling wines. • Wines discriminated according to aging duration but not dosage sugar type. • VIP scores generated from OPLS-DA identified potential aging markers. [ABSTRACT FROM AUTHOR]

Abstract Ethnopharmacological relevance Polygonum multiflorum Thund., a well-known and commonly-used TCM (Traditional Chinese Medicine) for treating hypertension, hyperlipidemia, premature graying of hair, and etc., has aroused wide concern for its reported potential liver toxicity. Due to its various active ingredients, the mechanisms underlying the hepatotoxicity of raw Polygonum multiflorum Thund (RPM) remain largely unknown. Aim of the study 1H NMR metabolomics was used to study the mechanism of RPM induced hepatotoxicity and disclosed the existence of hepatotoxicity and hepatoprotection conversion during RPM administration in mice. Materials and Methods Three dosages of RPM were administered by gavage to mice for consecutive 28 days. The serum and liver samples were collected and then subjected for histopathology observation, biochemical measurement and 1H NMR metabolic profiling. Results RPM caused oxidative stress and mitochondria dysfunction in mice, resulting in significant disturbance in energy metabolism, amino acid metabolism and pyrimidine metabolism and also inducing inflammatory responses. RPM induced hepatotoxicity in an apparent non-linear manner: the most severe in low dosage group, and to a less extent in medium group according to metabolomics analysis. The attenuation of liver injury in mice livers might result from the therapeutic effects, such as anti-oxidative capacity of RPM components. Conclusion RPM exerted a complicated non-linear manner in healthy recipients, switching between hepatoxicity and hepatoprotection dependent on the dosage and status of the body. Graphical abstract fx1 [ABSTRACT FROM AUTHOR]

The overall metabolic changes caused by cranberry juice or apple juice consumption using a global 1H NMR-based metabolomics approach were investigated. Eighteen female college students were given either cranberry or apple juice for three days using a cross-over design. Plasma and urine samples were collected and analyzed using 1H NMR-based metabolomics followed by multivariate analyses. No metabolic difference was observed in plasma before and after juice consumption. However, metabolome in plasma and urine after cranberry juice consumption were different from those after apple juice consumption. Cranberry juice consumption caused a greater increase in urinary excretion of hippuric acid and a higher level of citric acid in the plasma. Furthermore, cranberry juice decreased the plasma level of lactate, D-glucose, and two unidentified metabolites compared to apple juice consumption. The metabolomic changes caused by cranberry juice consumption may help to explain its reported health benefits.

[Display omitted] In the absence of appearance features, it is becoming increasingly difficult to accurately distinguish botanical herbal powders from different families or parts of the plant. In this study, we explored the biomarkers with neuroprotective effects in multi-parts ginseng based on metabolite content changes and biological functions. First, using 1H NMR and LC-MS metabolomics techniques and multivariate statistical analysis techniques to analyze the differential metabolites (DMs) in ginseng main root (TR), lateral root (LR) and Rhizome (RZ). Also, the analysis of 14 differential metabolites (DMs) using network pharmacology yielded 23 core targets associated with neuroprotective diseases, the KEGG pathway was enriched for 117 signaling pathways and 184 enrichment processes were shown in the GO analysis. Combined with the correlation analysis of primary differential metabolites (PDMs) and secondary differential metabolites (SDMs) content and network activity, we finally identified three biomarkers in different parts of ginseng, namely sucrose, ginsenoside Rb 1 , and ginsenoside Rb 3. Finally, the three biomarkers were validated by molecular docking and SH-SY5Y survival, and the results showed that sucrose, ginsenoside Rb 1 and ginsenoside Rb 3 exhibited favorable activity results in neuroprotective functions. The results of this study reveal the overall differences in the DMs of the three parts of ginseng, and three biomarkers can help identify different parts of ginseng powder, to avoid illegal substitution when ginseng is used in medicine. At the same time, it also provides a reference method for screening biomarkers that can distinguish different parts of traditional Chinese medicine. [ABSTRACT FROM AUTHOR]

Few comprehensive metabolic profiles of Lentinula edodes-derived polysaccharides on mammalians have been reported to date. In previous studies, a novel heteropolysaccharide, named L2, obtained from L. edodes had shown anti-ageing effects in mice. In this study, a 1H NMR-based metabolomics approach was used to investigate the urinary metabolome changes in aged mice gavaged with L2 for better understanding of anti-ageing mechanism. Six metabolites in urine were identified as potential markers for L2 treatment. An increase in α-oxoglutarate (α-OG) and a decrease in β-hydroxybutyrate, glutamate, glucose, choline, and lactate were observed. Enrichment pathway analysis indicated that 11 metabolic pathways were disrupted after L2 treatment. These altered metabolic pathways are mainly associated with energy metabolism, amino acid metabolism, and glycolysis. The results suggest that L2 could exert anti-ageing effects partly by regulating these pathways. Moreover, these findings may help better understand the mechanisms of the bioactivities of L. edodes-derived polysaccharides. Practical applications In this study, a 1H NMR-based metabolomics approach was used to investigate the urinary metabolome changes in aged mice gavaged with a novel heteropolysaccharide L2 from Lentinula edodes. The findings may help better understand the mechanisms of bioactivities of L. edodes-derived polysaccharides, and used for further development of L2 in food and medicine. [ABSTRACT FROM AUTHOR]

Ipomoea aquaticais an aquatic plant that is widely consumed in Southeast Asia as a vegetable. In this study, the influence of various ethanol ratios (0, 20, 50, 80, and 100%) as an extraction solvent and different drying methods including air drying, sun drying, and oven drying on phytochemical constituents ofI. aquaticawas investigated using a proton nuclear magnetic resonance-based metabolomics approach. The effect on α-glucosidase inhibitory activity and total phenolic content was also examined. Clear discrimination was observed between different ethanol ratios and different drying processes by principal component analysis. The highest α-glucosidase inhibitory activity was observed for absolute ethanol extract from the oven drying method with IC50value of 204.0 ± 59.0 µg/mL and total phenolic content value of 22.0 ± 0.7 µg gallic acid equivalent/mg extract. Correlation between the α-glucosidase inhibitory activity and the metabolite were analyzed using a partial least square analysis. The metabolites that are responsible for the activity were quercetin derivatives, chlorogenic acid derivatives, sucrose, and fructose. This study highlights the basis for future investigations ofI. aquaticaas a source of food that has the potential for nutraceutical enhancement and as ingredient in medicinal preparation. [ABSTRACT FROM PUBLISHER]

Metabolomic profiling is an increasingly important method for identifying potential biomarkers in cancer cells with a view towards improved diagnosis and treatment. Nuclear magnetic resonance (NMR) provides a potentially noninvasive means to accurately characterize differences in the metabolomic profiles of cells. In this work, we use 1H NMR to measure the metabolomic profiles of water soluble metabolites extracted from isogenic control and oncogenic HRAS-, KRAS-, and NRAS-transduced BEAS2B lung epithelial cells to determine the robustness of NMR metabolomic profiling in detecting differences between the transformed cells and their untransformed counterparts as well as differences among the RAS-transformed cells. Unique metabolomic signatures between control and RAS-transformed cell lines as well as among the three RAS isoform-transformed lines were found by applying principal component analysis to the NMR data. This study provides a proof of principle demonstration that NMR-based metabolomic profiling can robustly distinguish untransformed and RAS-transformed cells as well as cells transformed with different RAS oncogenic isoforms. Thus, our data may potentially provide new diagnostic signatures for RAS-transformed cells.

The Baltic Sea is one of the world's most pollution-threatened brackish environments and limited direct consumption of fatty fish from the Baltic Sea is recommended. The use of decontaminated Baltic Sea fish raw materials as fish feed could be a strategy to recycle Baltic Sea nutrients back into food chain, while relieving pressure on aqua-feed in the growing aquaculture industry. In this study, defatted fishmeal and semi-purified fish oil from the Baltic Sea were used in fish feeds for Arctic char ( Salvelinus alpinus L.). The effects of the Baltic Sea-sourced fish feeds on flesh lipid quality and fish metabolomics, compared with a standard commercial feed as a control, were determined. 1H NMR-based metabolomics studies indicated disturbances in energy metabolism and hepatic toxicity in fish fed both crude fishmeal and crude fish oil, associated with up-regulation (IGF-I, GHR-I, PPARα, PPARβ1A) and down-regulation (SREBP-1 and FAS) of hepatic genes expression. The content of n-3 long chain polyunsaturated fatty acids was not affected by the decontamination process. Thus, this short-term study demonstrates that decontaminating Baltic Sea-sourced fishmeal and fish oil reduces adverse effects in Arctic char. Practical applications: Decontaminated fish materials from the Baltic Sea were shown to be promising feed ingredients for Arctic char ( Salvelinus alpinus L.) compared with untreated Baltic Sea-sourced fish feed, which induced changes in fish physiology associated with energy metabolism and hepatotoxicity. Baltic Sea-sourced fish materials containing high levels of long chain polyunsaturated fatty acids are valuable feed ingredients. The Baltic Sea-sourced fishmeal (CFM and DFM) and fish oil (CFO and SPFO) were tested in fish feeds for Arctic charr ( Salvelinus alpinus), compared with a commercial diet as a control. Flesh lipid quality, metabolic profile in liver and muscle and hepatic gene expression (IGF-I, GHR-I, SREBP-1, etc.) were evaluated. [ABSTRACT FROM AUTHOR]

The effect of nonacylated and acylated anthocyanins on urinary metabolites in diabetic rats was investigated. Nonacylated anthocyanins extract from bilberries (NAAB) or acylated anthocyanins extract from purple potatoes (AAPP) was given to Zucker diabetic fatty (ZDF) rats for 8 weeks at daily doses of 25 and 50 mg/kg body weight. 1 H NMR metabolomics was applied to study alterations in urinary metabolites from three time points (weeks 1, 4, and 8). Both types of anthocyanins modulated the metabolites associated with the tricarboxylic acid cycle, gut microbiota metabolism, and renal function at weeks 1 and 4, such as 2-oxoglutarate, fumarate, alanine, trigonelline, and hippurate. In addition, only a high dose of AAPP decreased monosaccharides, formate, lactate, and glucose levels at week 4, suggesting improvement in energy production in mitochondria, glucose homeostasis, and oxidative stress. This study suggested different impacts of AAPP and NAAB on the metabolic profile of urine in diabetes.

The pharmacodynamics, 1H NMR metabolomics and endogenous network pharmacology strategy approaches were integrated to investigate the preventive mechanism of Gushudan (GSD) on kidney-yang-deficiency-syndrome (KYDS) rats in this study. Firstly, the KYDS rat model was achieved by hydrocortisone induction, and the efficacy of GSD on KYDS model rats was assessed by the pharmacodynamic indicators. Next, the comprehensive untargeted serum metabolic profile of rats was obtained in 1H NMR metabolomics study, 29 potential biomarkers closely associated with KYDS were identified, which were mainly involved in carbohydrate metabolism, amino acid metabolism and intestinal flora metabolism. In addition, the potential biomarkers-targets-pathways-disease metabolic network was further investigated for deeper understanding the preventive effects of GSD on KYDS rats and its mechanism, which was further obtained for the important targets related to biomarkers and diseases such as NOS3, PTGS2 and CXCL8, and important metabolic pathways such as glyoxylate and dicarboxylate metabolism, arginine and proline metabolism, and microbial metabolism in diverse environments. Finally, compared with our previous anti-osteoporosis study of GSD, it suggested that some similar metabolic pathways, which would provide some scientific reference of the existence of the kidney-bone axis under the traditional Chinese medicine (TCM) theory of "kidney dominates bone". [Display omitted] • A comprehensive approach of pharmacodynamics, metabolomics and endogenous network pharmacology were integrated. • Gushudan (GSD) has a preventive effect on kidney-yang-deficiency-syndrome (KYDS) rats. • Eleven new potential biomarkers were identified. • Carbohydrate metabolism, amino acid metabolism and intestinal flora metabolism were discovered. • An endogenous network pharmacology strategy to link the potential biomarkers to KYDS disease. [ABSTRACT FROM AUTHOR]

We evaluated the correlation between soil organic carbon (OC) content and metabolic responses of Eisenia fetida earthworms after exposure to phenanthrene (58 ± 3 mg/kg) spiked into seven artificial soils with OC contents ranging from 1 to 27% OC. Principal component analysis of 1H nuclear magnetic resonance (NMR) spectra of aqueous extracts identified statistically significant differences in the metabolic profiles of control and phenanthrene-exposed E. fetida in the 1% OC soil only. Partial least squares analysis identified a metabolic response in the four soils with OC values ≤11% which was well correlated to estimated phenanthrene porewater concentrations. The results suggest that the higher sorption capability of high OC soils decreased the bioavailability of phenanthrene and the subsequent metabolic response of E. fetida. [Copyright &y& Elsevier]

This study evaluated the cytotoxic activity of Tamoxifen (TMX), an anti-estrogen drug, with microalgal crude extracts (MCEs) in single and synergistic application (TMX-MCEs) on MCF-7 and 4T1 breast cancer cells, and non-cancerous Vero cells. The MCEs of Nannochloropsis oculata , Tetraselmis suecica and Chlorella sp. from five different solvents (methanol, MET; ethanol, ETH; water, W; chloroform, CHL; and hexane, HEX) were developed. The TMX-MCEs-ETH and W at the 1:2 and 1:3 ratios, attained IC 50 of 15.84-29.51 μg/mL against MCF-7; 13.8-31.62 μg/mL against 4T1; and 24.54-85.11 μg/mL against Vero cells. Higher late apoptosis was exhibited against MCF-7 by the TMX- N. oculata -ETH (41.15 %); and by the TMX- T. suecica -ETH (65.69 %) against 4T1 cells. The TMX- T. suecica -ETH also showed higher ADP/ATP ratios, but comparable Caspase activities to control. For Vero cells, overall apoptotic effects were lowered with synergistic application, and only early apoptosis was higher with TMX- T. suecica -ETH but at lower levels (29.84 %). The MCEs-W showed the presence of alanine, oleic acid, linoleic acid, lactic acid, and fumaric acid. Based on Principal Component Analysis (PCA), the spectral signals for polar solvents such as MET and ETH, were found in the same cluster, while the non-polar solvent CHL was with HEX, suggesting similar chemical profiles clustered for the same polarity. The CHL and HEX were more effective with N. oculata and T. suecica which were of the marine origin, while the ETH and MET were more effective with Chlorella sp., which was of the freshwater origin. The synergistic application of microalgal bioactive compounds with TMX can maintain the cytotoxicity against breast cancer cells whilst reducing the toxicity against non-cancerous Vero cells. These findings will benefit the biopharmaceutical, and functional and healthy food industries., Competing Interests: The authors declare no conflict of interest., (© 2022 The Author(s).)

• Metabolite-bioactivity correlation of Lupinus albus fractions were evaluated. • Chloroform fraction was identified as bioactive fraction. • 35 metabolites were identified from L. albus using 1H NMR metabolomics. • A total of 21 metabolites are tentatively identified using UHPLC-ESI-MS. Lupinus albus or white lupine has recently received increase attention for its medicinal values. Several studies have described the hypoglycemic effect of the white lupine, which is known as a food plant with potential value for treatment of diabetes. This study provides useful information for the identification and quantification of compounds in L. albus fractions by proton nuclear magnetic resonance (1H NMR) spectroscopy. In total, 35 metabolites were identified from L. albus fractions. Principal component analysis (PCA) was used as a multivariate projection method for visualizing the different composition of four different fractions. The bioactivities of fractions with different polarity obtained from the extract of L. albus seeds are reported. Among the fractions studied, the chloroform fraction (CF) exhibits a high free radical scavenging (DPPH) and α-glucosidase inhibitory activities with IC 50 values of 24.08 and 20.08 μg/mL, respectively. A partial least-squares analyses (PLS) model had been successfully performed to correlate the potential active metabolites with the corresponding biological activities. Metabolites containing proline, caprate, asparagine, lupinoisolone C, hydroxyiso lupalbigenin and some unknown compounds show high correlation with the bioactivities studied. Moreover, the structural identification in the active fraction was supported by ultrahigh-performance-liquid chromatography–electrospray ionization tandem mass spectrometry (UHPLC-ESI-MS/MS) analysis. A total of 21 metabolites were tentatively identified from MS/MS data by comparison with previously reported data. Most of these compounds are isoflavonoids without known biological activity. This information may be useful for developing functional food from L. albus with potential application in the management of diabetes. [ABSTRACT FROM AUTHOR]

1H NMR metabolomics profiles (A) of nattokinase-rich soymilk fermented by B. subtilis strain BSNK-5 at different fermentation time determined 52 metabolites. Combined with multivariate statistical analysis such as PCA(B-a), PLS-DA(B-b), OPLS-DA(C), the results showed the changes of molecular composition consistent with the prolongation of fermentation process and the 8 potential metabolic pathways (D) involved in the main active molecules. These findings indicated that 44 differential metabolites were identified which related to the changes of flavor, nutrition and function of fermented soymilk. BSNK-5 fermentation increased the synthesis of FAAs and some characteristic functional components, such as taurine, GABA and genistein, but also made the flavor of fermented soymilk worse. • NMR-based metabolomics of soymilk fermentation by B. subtilis was undertaken. • Compared with unfermented soymilk, 44 differential metabolites were identified. • Differential metabolites related to the changes of flavor, nutrition and function. • Eight key metabolic pathways were involved in the process of BSNK-5 fermentation. Microbial fermentation can endow food with unique flavors, increase its nutritional value and enhance functional characteristics. Our previous research has shown that liquid fermentation of soymilk by Bacillus subtilis BSNK-5 imparted new functional properties of to the fermented product via production of nattokinase. In this study, in order to further investigate the changes in the flavor, nutritional quality and functional characteristics of soymilk during fermentation using proton nuclear magnetic resonance (1H NMR) metabolomics to monitor the metabolite profile of BSNK-5-fermented soymilk. A total of 44 differential metabolites were identified between BSNK-5-fermented soymilk and uninoculated/unfermented soymilk, among which the levels of flavor-related substances (acetate, isovalerate and 2-methylbutyrate), nutrient-related substances (12 free amino acids), and functional substances (taurine, GABA and genistein) significantly increased after fermentation. These metabolites were closely associated with eight potential metabolic pathways. This work highlighted the significance of BSNK-5 strain in improving the nutritional quality and functional characteristics of fermented soymilk; however, the use of the strain also caused flavor deterioration. This study lays a theoretical foundation for the improvement and development of fermented soy products via liquid fermentation with B. subtilis. [ABSTRACT FROM AUTHOR]

Psoriasis is an inflammatory disease of the epidermis based on an immunological mechanism involving Langerhans cells and T lymphocytes that produce pro-inflammatory cytokines. Genetic factors, environmental factors, and improper nutrition are considered triggers of the disease. Numerous studies have reported that in a high number of patients, psoriasis is associated with obesity. Excess adipose tissue, typical of obesity, causes a systemic inflammatory status coming from the inflammatory active adipose tissue; therefore, weight reduction is a strategy to fight this pro-inflammatory state. This study aimed to evaluate how a nutritional regimen based on a ketogenic diet influenced the clinical parameters, metabolic profile, and inflammatory state of psoriasis patients. To this end, 30 psoriasis patients were subjected to a ketogenic nutritional regimen and monitored for 4 weeks by evaluating the clinical data, biochemical and clinical parameters, NMR metabolomic profile, and IL-2, IL-1β, TNF-α, IFN-γ, and IL-4 concentrations before and after the nutritional regimen. Our data show that a low-calorie ketogenic diet can be considered a successful strategy and therapeutic option to gain an improvement in psoriasis-related dysmetabolism, with significant correction of the full metabolic and inflammatory status.

Metabolomic profiling is an increasingly important method for identifying potential biomarkers in cancer cells with a view towards improved diagnosis and treatment. Nuclear magnetic resonance (NMR) provides a potentially noninvasive means to accurately characterize differences in the metabolomic profiles of cells. In this work, we use (1)H NMR to measure the metabolomic profiles of water soluble metabolites extracted from isogenic control and oncogenic HRAS-, KRAS-, and NRAS-transduced BEAS2B lung epithelial cells to determine the robustness of NMR metabolomic profiling in detecting differences between the transformed cells and their untransformed counterparts as well as differences among the RAS-transformed cells. Unique metabolomic signatures between control and RAS-transformed cell lines as well as among the three RAS isoform-transformed lines were found by applying principal component analysis to the NMR data. This study provides a proof of principle demonstration that NMR-based metabolomic profiling can robustly distinguish untransformed and RAS-transformed cells as well as cells transformed with different RAS oncogenic isoforms. Thus, our data may potentially provide new diagnostic signatures for RAS-transformed cells.

Due to the potential risk of accidental exposure to gamma radiation, it's critical to identify the biomarkers of radiation exposed creatures. In the present study, NMR based metabolomics combined with multivariate data analysis to evaluate the metabolites changed in the C57BL/6 mouse spleen after 4 days whole body exposure to 3.0 Gy and 7.8 Gy gamma radiations. Principal component analysis (PCA) and orthogonal projection to latent structures analysis (OPLS) are employed for classification and identification potential biomarkers associated with gamma irradiation. Two different strategies for NMR spectral data reduction (i.e., spectral binning and spectral deconvolution) are combined with normalize to constant sum and unit weight before multivariate data analysis, respectively. The combination of spectral deconvolution and normalization to unit weight is the best way for identifying discriminatory metabolites between the irradiation and control groups. Normalized to the constant sum may achieve some pseudo biomarkers. PCA and OPLS results shown that the exposed groups can be well separated from the control group. Leucine, 2-aminobutyrate, valine, lactate, arginine, glutathione, 2-oxoglutarate, creatine, tyrosine, phenylalanine, π-methylhistidine, taurine, myoinositol, glycerol and uracil are significantly elevated while ADP is decreased significantly. These significantly changed metabolites are associated with multiple metabolic pathways and may be potential biomarkers in the spleen exposed to gamma irradiation.

Background: The 5-year survival rate of the oral cancer patients has remained at approximately the 50% level during the past 30 years, possibly due to the poor tumor-selectivity of conventional anticancer drugs. This prompted us to search new plant extracts that have higher cytotoxicity against cancer cells than normal cells., Materials and Methods: Two human oral squamous cell carcinoma cell lines (HSC-2 and HSC-4) and two normal oral cells (gingival and periodontal ligament fibroblasts; HGF and HPLF) were incubated for 48 h with various concentrations of crude plant extract and the viable cell number was determined by the (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method. The 50% cytotoxic concentration (CC50) was determined from the dose-response curve. Tumor-specificity (TS) was determined by the following equation: TS=mean CC50 (normal cells)/mean CC50 (cancer cell lines). Metabolic profiling techniques based on (1)H nuclear magnetic resonance (NMR) were applied to gain the chemical structural insight for cytotoxicity induction., Results: Among 24 plant extracts, Camptotheca acuminate leaf, a well-known source for camptothecin, showed the highest TS value (88.3), followed by Vitis s.p.p. (>3.5), Sasa veitchii (>2.3) and Phellodendron amurense (>2.1), whereas other plant extracts showed much lower TS value (<2). These cytotoxic extracts made cluster on principal component analysis (PCA) score plot., Conclusion: The TS value determined by the present method seems to reflect the anti-tumor potential of each plant extract, while a part of the cytotoxic compounds present in these extracts may have common chemical structures., (Copyright© 2014 International Institute of Anticancer Research (Dr. John G. Delinassios), All rights reserved.)

Melanoma is a malignant tumor of melanocytes with high capability of invasion and rapid metastasis to other organs. Malignant melanoma is the most common metastatic malignancy found in Gastrointestinal Tract (GI). In this work, the 1H NMR-based metabolomics approach is used to investigate the metabolite profile differences of stomach tissue extracts of metastatic B16-F10 melanoma and control groups in C57BL/6J mouse and to search for specific metabolite biomarker candidates. Principal Component Analysis (PCA), an unsupervised multivariate data analysis method, is used to detect possible outliers, while Orthogonal Projection to Latent Structure (OPLS), a supervised multivariate data analysis method, is employed to evaluate important metabolites responsible for discriminating the control and the melanoma groups. Both PCA and OPLS results reveal that the melanoma group can be well separated from its control group. Among the 50 identified metabolites, it is found that the concentrations of 19 metabolites are significantly changed with the levels of O-phosphocholine and hypoxanthine down-regulated while the levels of isoleucine, leucine, valine, isobutyrate, threonine, cadaverine, alanine, glutamate, glutamine, methionine, citrate, asparagine, tryptophan, glycine, serine, uracil, and formate up-regulated in the melanoma group. These significantly changed metabolites are associated with multiple biological pathways and may be potential biomarkers for metastatic melanoma in stomach.