Untargeted metabolomics aims to gather information on as many metabolites as possible in biologic... more Untargeted metabolomics aims to gather information on as many metabolites as possible in biological systems by taking into account all information present in the data sets. Here we describe a detailed protocol for large-scale untargeted metabolomics of plant tissues, based on reversed phase liquid chromatography coupled to high-resolution mass spectrometry (LC-QTOF MS) of aqueous methanol extracts. Dedicated software, MetAlign, is used for automated baseline correction and alignment of all extracted mass peaks across all samples, producing detailed information on the relative abundance of thousands of mass signals representing hundreds of metabolites. Subsequent statistics and bioinformatics tools can be used to provide a detailed view on the differences and similarities between (groups of) samples or to link metabolomics data to other systems biology information, genetic markers and/or specific quality parameters. The complete procedure from metabolite extraction to assembly of a data matrix with aligned mass signal intensities takes about 6 days for 50 samples.
Methods in molecular biology (Clifton, N.J.), 2012
The identification of metabolites in biochemical studies is a major bottleneck in the proliferati... more The identification of metabolites in biochemical studies is a major bottleneck in the proliferating field of metabolomics. In particular in plant metabolomics, given the diversity and abundance of endogenous secondary metabolites in plants, the identification of these is not only challenging but also essential to understanding their biological role in the plant, and their value to quality and nutritional attributes as food crops. With the new generation of analytical technologies, in which liquid chromatography (LC)-mass spectrometry (MS) and nuclear magnetic resonance (NMR) play a pioneering role, profiling metabolites in complex extracts is feasible at high throughput. However, the identification of key metabolites remains a limitation given the analytical effort necessary for traditional structural elucidation strategies. The hyphenation of LC-solid phase extraction (SPE)-NMR is a powerful analytical platform for isolating and concentrating metabolites for unequivocal identificat...
Se pu = Chinese journal of chromatography / Zhongguo hua xue hui, 2005
As one of the main phytoalexins and phytoestrogens, glyceollin is an important prenylflavonoid in... more As one of the main phytoalexins and phytoestrogens, glyceollin is an important prenylflavonoid in Glycine max [L.] Merri. (soybean). Many kinds of elicitors can be used to induce its accumulation. Its biosynthesis pathway is commonly used to study the characteristics of prenyltransferase, which catalyzes the prenylated reaction happening in a very few plant families in nature. Glycinol, the direct precursor of glyceollin, is necessary to study the prenylated reaction in soybean. In comparing with the other elicitors to elicit the glycinol accumulation in soybean cotyledons, AgNO3 is the most effective elicitor. Exposure of 6-8 days old cotyledons to 0.01 mol/L AgNO3 and incubation for 24 h result in the accumulation of 256 microg (glycinol)/g (fresh weight). The glycinol was extracted by methanol. Then the isolation and purification were conducted by preparative high performance liquid chromatography. Instead of 100% acetonitrile-0.1% formic acid as the elution system, the extract w...
Metabolomics studies rely on the analysis of the multitude of small molecules (metabolites) prese... more Metabolomics studies rely on the analysis of the multitude of small molecules (metabolites) present in a biological system. Most commonly, metabolomics is heavily supported by mass spectrometry (MS) and nuclear magnetic resonance (NMR) as parallel technologies that provide an overview of the metabolome and high-power compound elucidation. Over and above large-scale analysis, a major effort is needed for unequivocal identification of metabolites. The combination of liquid chromatography (LC)-MS and NMR is a powerful methodology for identifying metabolites. Better chemical characterization of the metabolome will undoubtedly enlarge knowledge of any biological system.
Untargeted metabolomics aims to gather information on as many metabolites as possible in biologic... more Untargeted metabolomics aims to gather information on as many metabolites as possible in biological systems by taking into account all information present in the data sets. Here we describe a detailed protocol for large-scale untargeted metabolomics of plant tissues, based on reversed phase liquid chromatography coupled to high-resolution mass spectrometry (LC-QTOF MS) of aqueous methanol extracts. Dedicated software, MetAlign, is used for automated baseline correction and alignment of all extracted mass peaks across all samples, producing detailed information on the relative abundance of thousands of mass signals representing hundreds of metabolites. Subsequent statistics and bioinformatics tools can be used to provide a detailed view on the differences and similarities between (groups of) samples or to link metabolomics data to other systems biology information, genetic markers and/or specific quality parameters. The complete procedure from metabolite extraction to assembly of a data matrix with aligned mass signal intensities takes about 6 days for 50 samples.
Nuclear magnetic resonance (NMR) and liquid chromatography-mass spectrometry (LCMS) are frequentl... more Nuclear magnetic resonance (NMR) and liquid chromatography-mass spectrometry (LCMS) are frequently used as technological platforms for metabolomics applications. In this study, the metabolic profiles of ripe fruits from 50 different tomato cultivars, including beef, cherry and round types, were recorded by both 1 H NMR and accurate mass LC-quadrupole time-of-flight (QTOF) MS. Different analytical selectivities were found for these both profiling techniques. In fact, NMR and LCMS provided complementary data, as the metabolites detected belong to essentially different metabolic pathways. Yet, upon unsupervised multivariate analysis, both NMR and LCMS datasets revealed a clear segregation of, on the one hand, the cherry tomatoes and, on the other hand, the beef and round tomatoes. Intra-method (NMR-NMR, LCMS-LCMS) and inter-method (NMR-LCMS) correlation analyses were performed enabling the annotation of metabolites from highly correlating metabolite signals. Signals belonging to the same metabolite or to chemically related metabolites are among the highest correlations found. Inter-method correlation analysis produced highly informative and complementary information for the identification of metabolites, even in de case of low abundant NMR signals. The applied approach appears to be a promising strategy in extending the analytical capacities of these metabolomics techniques with regard to the discovery and identification of biomarkers and yet unknown metabolites.
Fruit maturation and tissue differentiation are important topics in plant physiology. These biolo... more Fruit maturation and tissue differentiation are important topics in plant physiology. These biological phenomena are accompanied by specific alterations in the biological system, such as differences in the type and concentration of metabolites. The secondary metabolism of tomato (Solanum lycopersicum) fruit was monitored by using liquid chromatography (LC) coupled to photo-diode array (PDA) detection, fluorescence detection (FD), and mass spectrometry (MS). Through this integrated approach different classes of compounds were analysed: carotenoids, xanthophylls, chlorophylls, tocopherols, ascorbic acid, flavonoids, phenolic acids, glycoalkaloids, saponins, and other glycosylated derivatives. Related metabolite profiles of peel and flesh were found between several commercial tomato cultivars indicating similar metabolite trends despite the genetic background. For a single tomato cultivar, metabolite profiles of different fruit tissues (vascular attachment region, columella and placenta, epidermis, pericarp, and jelly parenchyma) were examined at the green, breaker, turning, pink, and red stages of fruit development. Unrelated to the chemical nature of the metabolites, behavioural patterns could be assigned to specific ripening stages or tissues. These findings suggest spatio-temporal specificity in the accumulation of endogenous metabolites from tomato fruit.
Although the prevalence of main idiopathic forms of inflammatory bowel disease (IBD) has risen co... more Although the prevalence of main idiopathic forms of inflammatory bowel disease (IBD) has risen considerably over the last decades, their clinical features do not allow accurate prediction of prognosis, likelihood of disease progression, or response to specific therapy. Through a better understanding of the molecular pathways involved in IBD and the promise of more targeted therapies, the personalized approach to the management of IBD shows potential. To achieve this, there remains a significant need to better understand the disease process at cellular and molecular levels for any given individual with IBD. The complexity of biological functional networks behind the etiology of IBD highlights the need for their comprehensive analysis. In this, omics technologies can generate a systemic view of IBD pathogenesis on which to base novel, multiple pathway-integrated therapies. Omics sciences have just started to contribute here by generating gene, protein expression, metabolite data at gl...
Epidemiological studies have repeatedly found that whole-grain (WG) cereal foods reduce the risk ... more Epidemiological studies have repeatedly found that whole-grain (WG) cereal foods reduce the risk of several lifestyle-related diseases, though consistent clinical outcomes and mechanisms are elusive. To compare the effects of a WG-rich diet with a matched refinedgrain (RG) diet on plasma biomarkers and bowel health parameters, seventeen healthy subjects (eleven females and six males) completed an exploratory cross-over study with a 2-week intervention diet based on either WG-or RG-based foods, separated by a washout of at least 5 weeks. Both diets were the same except for the use of WG (150 g/d) or RG foods. Subjects undertook a 4 h postprandial challenge on day 8 of each intervention diet. After 2 weeks, the WG diet tended to decrease plasma total and LDL-cholesterol (both P¼0·09), but did not change plasma HDL-cholesterol, fasting glucose, C-reactive protein or homocysteine compared with the RG diet. Plasma betaine and alkylresorcinol concentrations were elevated after 1 week of the WG diet (P¼0·01 and P, 0·0001, respectively). Clostridium leptum populations in faeces were increased after the WG diet, along with a trend for decreased faecal water pH (P¼0·096) and increased stool frequency (P,0·0001) compared with the RG diet. A short controlled intervention trial with a variety of commercially available WG-based products tended to improve biomarkers of CVD compared with a RG diet. Changes in faecal microbiota related to increased fibre fermentation and increased plasma betaine concentrations point to both fibre and phytochemical components of WG being important in mediating any potential health effects.
Overall metabolic modifications between fruit of light-hyperresponsive highpigment ( hp ) tomato ... more Overall metabolic modifications between fruit of light-hyperresponsive highpigment ( hp ) tomato ( Lycopersicon esculentum ) mutant plants and isogenic nonmutant (wt) control plants were compared.
In any metabolomics experiment, robustness and reproducibility of data collection is of vital imp... more In any metabolomics experiment, robustness and reproducibility of data collection is of vital importance. These become more important in collaborative studies where data is to be collected on multiple instruments. With minimisation of variance in sample preparation and instrument performance it is possible to elucidate even subtle differences in metabolite fingerprints due to genotype or biological treatment. In this paper we report on an inter laboratory comparison of plant derived samples by [ 1 H]-NMR spectroscopy across five different sites and within those sites utilising instruments with different probes and magnetic field strengths of 9.4 T (400 MHz), 11.7 T (500 MHz) and 14.1 T (600 MHz). Whilst the focus of the study is on consistent data collection across laboratories, aspects of sample stability and the requirement for sample rotation within the NMR magnet are also discussed. Comparability of the datasets from participating laboratories was exceptionally good and the data were amenable to comparative analysis by multivariate statistics. Field strength differences can be adjusted for in the data pre-processing and multivariate analysis demonstrating that [ 1 H]-NMR fingerprinting is the ideal technique for large scale plant metabolomics data collection requiring the participation of multiple laboratories.
Identification and quantification of metabolites occurring within specific cell types or single c... more Identification and quantification of metabolites occurring within specific cell types or single cells of plants and other organisms is of particular interest for natural product chemistry, chemical ecology, and biochemistry in general. The integration of studies at the gene, transcript, protein and metabolite levels in localized regions will provide useful information for the understanding of biology as a system. In this review, we summarize the latest developments in the analysis of metabolites present in small samples, micrometabolomics, dealing with sample preparation methods, with focus on laser-assisted microdissection, and the analytical technologies used. Mass spectrometry (MS) and nuclear magnetic resonance (NMR) are among the most emergent technologies in metabolomics, enabling the shortest route toward metabolite identification.
neo-Clerodane diterpenes of plant origen are molecules difficult to monitor due to their nonspeci... more neo-Clerodane diterpenes of plant origen are molecules difficult to monitor due to their nonspecific UV/vis absorption. The present work describes for the first time the application of the LC-MS-SPE-NMR technique for the isolation and characterization of three new neo-clerodane diterpenes, 3beta-hydroxyteucroxylepin and teuluteumin A and teuluteumin B, from Teucrium luteum subsp. flavovirens, harvested from two different locations.
Visceral adiposity is increasingly recognized as a key condition for the development of obesity r... more Visceral adiposity is increasingly recognized as a key condition for the development of obesity related disorders, with the ratio between visceral adipose tissue (VAT) and subcutaneous adipose tissue (SAT) reported as the best correlate of cardiometabolic risk. In this study, using a cohort of 40 obese females (age: 25-45 y, BMI: 28-40 kg/m 2 ) under healthy clinical conditions and monitored over a 2 weeks period we examined the relationships between different body composition parameters, estimates of visceral adiposity and blood/urine metabolic profiles. Metabonomics and lipidomics analysis of blood plasma and urine were employed in combination with in vivo quantitation of body composition and abdominal fat distribution using iDXA and computerized tomography. Of the various visceral fat estimates, VAT/SAT and VAT/total abdominal fat ratios exhibited significant associations with regio-specific body lean and fat composition. The integration of these visceral fat estimates with metabolic profiles of blood and urine described a distinct amino acid, diacyl and ether phospholipid phenotype in women with higher visceral fat. Metabolites important in predicting visceral fat adiposity as assessed by Random forest analysis highlighted 7 most robust markers, including tyrosine, glutamine, :3 lipid species. Unexpectedly, the visceral fat associated inflammatory profiles were shown to be highly influenced by inter-days and between-subject variations. Nevertheless, the visceral fat associated amino acid and lipid signature is proposed to be further validated for future patient stratification and cardiometabolic health diagnostics.
Increasing evidence points toward the critical and long-term involvement of prenatal and early nu... more Increasing evidence points toward the critical and long-term involvement of prenatal and early nutrition and lifestyle on later health and disease risk predisposition. Metabolomics is now a well-established top-down systems biology approach that explores the genetic-environment-health paradigm. The generalization of such approaches has opened new research areas to deepen our current understanding of many physiological processes, as well as foods and nutrient functionalities in target populations. It is envisioned that this will provide new avenues toward preventive medicine and prognostic strategies for tailored therapeutic and personalized nutrition management. The development of systems biology approaches and the new generation of biomarker patterns will provide the opportunity to associate complex metabolic regulations with the etiology of multifactorial pediatric diseases. This may subsequently lead to the development of system mechanistic hypotheses that could be targeted with new nutritional personalized concepts. Therefore, this review aims to describe recent applications of metabolomics in preclinical and clinical fields with insights into disease diagnostics/monitoring and improvement of homeostasis metabolic regulation that may be translatable to novel therapeutic and nutrition advances in pediatric research.
Untargeted metabolomics aims to gather information on as many metabolites as possible in biologic... more Untargeted metabolomics aims to gather information on as many metabolites as possible in biological systems by taking into account all information present in the data sets. Here we describe a detailed protocol for large-scale untargeted metabolomics of plant tissues, based on reversed phase liquid chromatography coupled to high-resolution mass spectrometry (LC-QTOF MS) of aqueous methanol extracts. Dedicated software, MetAlign, is used for automated baseline correction and alignment of all extracted mass peaks across all samples, producing detailed information on the relative abundance of thousands of mass signals representing hundreds of metabolites. Subsequent statistics and bioinformatics tools can be used to provide a detailed view on the differences and similarities between (groups of) samples or to link metabolomics data to other systems biology information, genetic markers and/or specific quality parameters. The complete procedure from metabolite extraction to assembly of a data matrix with aligned mass signal intensities takes about 6 days for 50 samples.
Methods in molecular biology (Clifton, N.J.), 2012
The identification of metabolites in biochemical studies is a major bottleneck in the proliferati... more The identification of metabolites in biochemical studies is a major bottleneck in the proliferating field of metabolomics. In particular in plant metabolomics, given the diversity and abundance of endogenous secondary metabolites in plants, the identification of these is not only challenging but also essential to understanding their biological role in the plant, and their value to quality and nutritional attributes as food crops. With the new generation of analytical technologies, in which liquid chromatography (LC)-mass spectrometry (MS) and nuclear magnetic resonance (NMR) play a pioneering role, profiling metabolites in complex extracts is feasible at high throughput. However, the identification of key metabolites remains a limitation given the analytical effort necessary for traditional structural elucidation strategies. The hyphenation of LC-solid phase extraction (SPE)-NMR is a powerful analytical platform for isolating and concentrating metabolites for unequivocal identificat...
Se pu = Chinese journal of chromatography / Zhongguo hua xue hui, 2005
As one of the main phytoalexins and phytoestrogens, glyceollin is an important prenylflavonoid in... more As one of the main phytoalexins and phytoestrogens, glyceollin is an important prenylflavonoid in Glycine max [L.] Merri. (soybean). Many kinds of elicitors can be used to induce its accumulation. Its biosynthesis pathway is commonly used to study the characteristics of prenyltransferase, which catalyzes the prenylated reaction happening in a very few plant families in nature. Glycinol, the direct precursor of glyceollin, is necessary to study the prenylated reaction in soybean. In comparing with the other elicitors to elicit the glycinol accumulation in soybean cotyledons, AgNO3 is the most effective elicitor. Exposure of 6-8 days old cotyledons to 0.01 mol/L AgNO3 and incubation for 24 h result in the accumulation of 256 microg (glycinol)/g (fresh weight). The glycinol was extracted by methanol. Then the isolation and purification were conducted by preparative high performance liquid chromatography. Instead of 100% acetonitrile-0.1% formic acid as the elution system, the extract w...
Metabolomics studies rely on the analysis of the multitude of small molecules (metabolites) prese... more Metabolomics studies rely on the analysis of the multitude of small molecules (metabolites) present in a biological system. Most commonly, metabolomics is heavily supported by mass spectrometry (MS) and nuclear magnetic resonance (NMR) as parallel technologies that provide an overview of the metabolome and high-power compound elucidation. Over and above large-scale analysis, a major effort is needed for unequivocal identification of metabolites. The combination of liquid chromatography (LC)-MS and NMR is a powerful methodology for identifying metabolites. Better chemical characterization of the metabolome will undoubtedly enlarge knowledge of any biological system.
Untargeted metabolomics aims to gather information on as many metabolites as possible in biologic... more Untargeted metabolomics aims to gather information on as many metabolites as possible in biological systems by taking into account all information present in the data sets. Here we describe a detailed protocol for large-scale untargeted metabolomics of plant tissues, based on reversed phase liquid chromatography coupled to high-resolution mass spectrometry (LC-QTOF MS) of aqueous methanol extracts. Dedicated software, MetAlign, is used for automated baseline correction and alignment of all extracted mass peaks across all samples, producing detailed information on the relative abundance of thousands of mass signals representing hundreds of metabolites. Subsequent statistics and bioinformatics tools can be used to provide a detailed view on the differences and similarities between (groups of) samples or to link metabolomics data to other systems biology information, genetic markers and/or specific quality parameters. The complete procedure from metabolite extraction to assembly of a data matrix with aligned mass signal intensities takes about 6 days for 50 samples.
Nuclear magnetic resonance (NMR) and liquid chromatography-mass spectrometry (LCMS) are frequentl... more Nuclear magnetic resonance (NMR) and liquid chromatography-mass spectrometry (LCMS) are frequently used as technological platforms for metabolomics applications. In this study, the metabolic profiles of ripe fruits from 50 different tomato cultivars, including beef, cherry and round types, were recorded by both 1 H NMR and accurate mass LC-quadrupole time-of-flight (QTOF) MS. Different analytical selectivities were found for these both profiling techniques. In fact, NMR and LCMS provided complementary data, as the metabolites detected belong to essentially different metabolic pathways. Yet, upon unsupervised multivariate analysis, both NMR and LCMS datasets revealed a clear segregation of, on the one hand, the cherry tomatoes and, on the other hand, the beef and round tomatoes. Intra-method (NMR-NMR, LCMS-LCMS) and inter-method (NMR-LCMS) correlation analyses were performed enabling the annotation of metabolites from highly correlating metabolite signals. Signals belonging to the same metabolite or to chemically related metabolites are among the highest correlations found. Inter-method correlation analysis produced highly informative and complementary information for the identification of metabolites, even in de case of low abundant NMR signals. The applied approach appears to be a promising strategy in extending the analytical capacities of these metabolomics techniques with regard to the discovery and identification of biomarkers and yet unknown metabolites.
Fruit maturation and tissue differentiation are important topics in plant physiology. These biolo... more Fruit maturation and tissue differentiation are important topics in plant physiology. These biological phenomena are accompanied by specific alterations in the biological system, such as differences in the type and concentration of metabolites. The secondary metabolism of tomato (Solanum lycopersicum) fruit was monitored by using liquid chromatography (LC) coupled to photo-diode array (PDA) detection, fluorescence detection (FD), and mass spectrometry (MS). Through this integrated approach different classes of compounds were analysed: carotenoids, xanthophylls, chlorophylls, tocopherols, ascorbic acid, flavonoids, phenolic acids, glycoalkaloids, saponins, and other glycosylated derivatives. Related metabolite profiles of peel and flesh were found between several commercial tomato cultivars indicating similar metabolite trends despite the genetic background. For a single tomato cultivar, metabolite profiles of different fruit tissues (vascular attachment region, columella and placenta, epidermis, pericarp, and jelly parenchyma) were examined at the green, breaker, turning, pink, and red stages of fruit development. Unrelated to the chemical nature of the metabolites, behavioural patterns could be assigned to specific ripening stages or tissues. These findings suggest spatio-temporal specificity in the accumulation of endogenous metabolites from tomato fruit.
Although the prevalence of main idiopathic forms of inflammatory bowel disease (IBD) has risen co... more Although the prevalence of main idiopathic forms of inflammatory bowel disease (IBD) has risen considerably over the last decades, their clinical features do not allow accurate prediction of prognosis, likelihood of disease progression, or response to specific therapy. Through a better understanding of the molecular pathways involved in IBD and the promise of more targeted therapies, the personalized approach to the management of IBD shows potential. To achieve this, there remains a significant need to better understand the disease process at cellular and molecular levels for any given individual with IBD. The complexity of biological functional networks behind the etiology of IBD highlights the need for their comprehensive analysis. In this, omics technologies can generate a systemic view of IBD pathogenesis on which to base novel, multiple pathway-integrated therapies. Omics sciences have just started to contribute here by generating gene, protein expression, metabolite data at gl...
Epidemiological studies have repeatedly found that whole-grain (WG) cereal foods reduce the risk ... more Epidemiological studies have repeatedly found that whole-grain (WG) cereal foods reduce the risk of several lifestyle-related diseases, though consistent clinical outcomes and mechanisms are elusive. To compare the effects of a WG-rich diet with a matched refinedgrain (RG) diet on plasma biomarkers and bowel health parameters, seventeen healthy subjects (eleven females and six males) completed an exploratory cross-over study with a 2-week intervention diet based on either WG-or RG-based foods, separated by a washout of at least 5 weeks. Both diets were the same except for the use of WG (150 g/d) or RG foods. Subjects undertook a 4 h postprandial challenge on day 8 of each intervention diet. After 2 weeks, the WG diet tended to decrease plasma total and LDL-cholesterol (both P¼0·09), but did not change plasma HDL-cholesterol, fasting glucose, C-reactive protein or homocysteine compared with the RG diet. Plasma betaine and alkylresorcinol concentrations were elevated after 1 week of the WG diet (P¼0·01 and P, 0·0001, respectively). Clostridium leptum populations in faeces were increased after the WG diet, along with a trend for decreased faecal water pH (P¼0·096) and increased stool frequency (P,0·0001) compared with the RG diet. A short controlled intervention trial with a variety of commercially available WG-based products tended to improve biomarkers of CVD compared with a RG diet. Changes in faecal microbiota related to increased fibre fermentation and increased plasma betaine concentrations point to both fibre and phytochemical components of WG being important in mediating any potential health effects.
Overall metabolic modifications between fruit of light-hyperresponsive highpigment ( hp ) tomato ... more Overall metabolic modifications between fruit of light-hyperresponsive highpigment ( hp ) tomato ( Lycopersicon esculentum ) mutant plants and isogenic nonmutant (wt) control plants were compared.
In any metabolomics experiment, robustness and reproducibility of data collection is of vital imp... more In any metabolomics experiment, robustness and reproducibility of data collection is of vital importance. These become more important in collaborative studies where data is to be collected on multiple instruments. With minimisation of variance in sample preparation and instrument performance it is possible to elucidate even subtle differences in metabolite fingerprints due to genotype or biological treatment. In this paper we report on an inter laboratory comparison of plant derived samples by [ 1 H]-NMR spectroscopy across five different sites and within those sites utilising instruments with different probes and magnetic field strengths of 9.4 T (400 MHz), 11.7 T (500 MHz) and 14.1 T (600 MHz). Whilst the focus of the study is on consistent data collection across laboratories, aspects of sample stability and the requirement for sample rotation within the NMR magnet are also discussed. Comparability of the datasets from participating laboratories was exceptionally good and the data were amenable to comparative analysis by multivariate statistics. Field strength differences can be adjusted for in the data pre-processing and multivariate analysis demonstrating that [ 1 H]-NMR fingerprinting is the ideal technique for large scale plant metabolomics data collection requiring the participation of multiple laboratories.
Identification and quantification of metabolites occurring within specific cell types or single c... more Identification and quantification of metabolites occurring within specific cell types or single cells of plants and other organisms is of particular interest for natural product chemistry, chemical ecology, and biochemistry in general. The integration of studies at the gene, transcript, protein and metabolite levels in localized regions will provide useful information for the understanding of biology as a system. In this review, we summarize the latest developments in the analysis of metabolites present in small samples, micrometabolomics, dealing with sample preparation methods, with focus on laser-assisted microdissection, and the analytical technologies used. Mass spectrometry (MS) and nuclear magnetic resonance (NMR) are among the most emergent technologies in metabolomics, enabling the shortest route toward metabolite identification.
neo-Clerodane diterpenes of plant origen are molecules difficult to monitor due to their nonspeci... more neo-Clerodane diterpenes of plant origen are molecules difficult to monitor due to their nonspecific UV/vis absorption. The present work describes for the first time the application of the LC-MS-SPE-NMR technique for the isolation and characterization of three new neo-clerodane diterpenes, 3beta-hydroxyteucroxylepin and teuluteumin A and teuluteumin B, from Teucrium luteum subsp. flavovirens, harvested from two different locations.
Visceral adiposity is increasingly recognized as a key condition for the development of obesity r... more Visceral adiposity is increasingly recognized as a key condition for the development of obesity related disorders, with the ratio between visceral adipose tissue (VAT) and subcutaneous adipose tissue (SAT) reported as the best correlate of cardiometabolic risk. In this study, using a cohort of 40 obese females (age: 25-45 y, BMI: 28-40 kg/m 2 ) under healthy clinical conditions and monitored over a 2 weeks period we examined the relationships between different body composition parameters, estimates of visceral adiposity and blood/urine metabolic profiles. Metabonomics and lipidomics analysis of blood plasma and urine were employed in combination with in vivo quantitation of body composition and abdominal fat distribution using iDXA and computerized tomography. Of the various visceral fat estimates, VAT/SAT and VAT/total abdominal fat ratios exhibited significant associations with regio-specific body lean and fat composition. The integration of these visceral fat estimates with metabolic profiles of blood and urine described a distinct amino acid, diacyl and ether phospholipid phenotype in women with higher visceral fat. Metabolites important in predicting visceral fat adiposity as assessed by Random forest analysis highlighted 7 most robust markers, including tyrosine, glutamine, :3 lipid species. Unexpectedly, the visceral fat associated inflammatory profiles were shown to be highly influenced by inter-days and between-subject variations. Nevertheless, the visceral fat associated amino acid and lipid signature is proposed to be further validated for future patient stratification and cardiometabolic health diagnostics.
Increasing evidence points toward the critical and long-term involvement of prenatal and early nu... more Increasing evidence points toward the critical and long-term involvement of prenatal and early nutrition and lifestyle on later health and disease risk predisposition. Metabolomics is now a well-established top-down systems biology approach that explores the genetic-environment-health paradigm. The generalization of such approaches has opened new research areas to deepen our current understanding of many physiological processes, as well as foods and nutrient functionalities in target populations. It is envisioned that this will provide new avenues toward preventive medicine and prognostic strategies for tailored therapeutic and personalized nutrition management. The development of systems biology approaches and the new generation of biomarker patterns will provide the opportunity to associate complex metabolic regulations with the etiology of multifactorial pediatric diseases. This may subsequently lead to the development of system mechanistic hypotheses that could be targeted with new nutritional personalized concepts. Therefore, this review aims to describe recent applications of metabolomics in preclinical and clinical fields with insights into disease diagnostics/monitoring and improvement of homeostasis metabolic regulation that may be translatable to novel therapeutic and nutrition advances in pediatric research.
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Papers by Sofia Moco