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1.
Cell Biochem Funct ; 37(6): 408-423, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31328813

RESUMO

Current understanding of vitamin D tends to be focussed on the measurement of the major circulating form 25-hydroxyvitamin D3 (25OHD3) and its conversion to the active hormonal form, 1α,25-dihydroxyvitamin D3 (1α,25(OH)2 D3) via the enzyme 25-hydroxyvitamin D-1α-hydroxylase (CYP27B1). However, whilst these metabolites form the endocrine backbone of vitamin D physiology, it is important to recognise that there are other metabolic and catabolic pathways that are now recognised as being crucially important to vitamin D function. These pathways include C3-epimerization, CYP24A1 hydroxylase, CYP11A1 alternative metabolism of vitamin D3, and phase II metabolism. Endogenous metabolites beyond 25OHD3 are usually present at low endogenous levels and may only be functional in specific target tissues rather than in the general circulation. However, the technologies available to measure these metabolites have also improved, so that measurement of alternative vitamin D metabolic pathways may become more routine in the near future. The aim of this review is to provide a comprehensive overview of the various pathways of vitamin D metabolism, as well as describe the analytical techniques currently available to measure these vitamin D metabolites.


Assuntos
Metaboloma/fisiologia , Vitamina D/análise , Vitamina D/metabolismo , Animais , Humanos
2.
Food Chem Toxicol ; 131: 110547, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31170423

RESUMO

Safety concerns arising from the consumption of foods derived from genetically modified (GM) crops remains a controversial subject. We report here a faecal microbiota compositional analysis in Wistar rats from the GMO90 + study, which fed glyphosate-tolerant NK603 (+/- Roundup application) and Bt toxin MON810 GM maize for 6 months in comparison to their closest non-GM isogenic lines. We first integrated the faecal microbiota compositional data with results from plasma metabolomics to understand which bacterial species can influence host metabolism. Coriobacteriaceae and Acetatifactor significantly predicted plasma metabolic profile in males, while Bifidobacterium and Ruminococcus were able to predict female plasma metabolites. We then investigated the differences in fecal microbiota composition between group of rats fed MON810 or NK603 GM maize in comparison to their isogenic lines. Bacterial community richness was not altered by the test diets. There were no statistically significant differences in taxa abundance in the rat faecal microbiota that we could attribute to the consumption of either MON810 or NK603. We show that the consumption of the widely cultivated GM maize varieties NK603 and MON810 even up to 33% of the total diet had no effect on the status of the faecal microbiota compared to non-GM near isogenic lines.


Assuntos
Fezes/microbiologia , Alimentos Geneticamente Modificados , Microbioma Gastrointestinal/fisiologia , Metaboloma/fisiologia , Plantas Geneticamente Modificadas , Zea mays , Animais , Dieta , Feminino , Masculino , RNA Bacteriano/análise , RNA Ribossômico 16S/análise , Ratos Wistar
3.
Mar Drugs ; 17(5)2019 May 18.
Artigo em Inglês | MEDLINE | ID: mdl-31109094

RESUMO

The mass spectrometry-based metabolomics approach has become a powerful tool for the quantitative analysis of small-molecule metabolites in biological samples. Nostoc flagelliforme, an edible cyanobacterium with herbal value, serves as an unexploited bioresource for small molecules. In natural environments, N. flagelliforme undergoes repeated cycles of rehydration and dehydration, which are interrupted by either long- or short-term dormancy. In this study, we performed an untargeted metabolite profiling of N. flagelliforme samples at three physiological states: Dormant (S1), physiologically fully recovered after rehydration (S2), and physiologically partially inhibited following dehydration (S3). Significant metabolome differences were identified based on the OPLS-DA (orthogonal projections to latent structures discriminant analysis) model. In total, 183 differential metabolites (95 up-regulated; 88 down-regulated) were found during the rehydration process (S2 vs. S1), and 130 (seven up-regulated; 123 down-regulated) during the dehydration process (S3 vs. S2). Thus, it seemed that the metabolites' biosynthesis mainly took place in the rehydration process while the degradation or possible conversion occurred in the dehydration process. In addition, lipid profile differences were particularly prominent, implying profound membrane phase changes during the rehydration-dehydration cycle. In general, this study expands our understanding of the metabolite dynamics in N. flagelliforme and provides biotechnological clues for achieving the efficient production of those metabolites with medical potential.


Assuntos
Dessecação , Regulação Bacteriana da Expressão Gênica/fisiologia , Metaboloma/fisiologia , Nostoc/química , Membrana Celular/genética , Membrana Celular/metabolismo , Meio Ambiente , Nostoc/metabolismo , Água
4.
Artigo em Inglês | MEDLINE | ID: mdl-31108321

RESUMO

Urinary metabolomics offers a non-invasive means of obtaining information about the system-wide biological health of a patient. Untargeted metabolomics approaches using one-dimensional gas chromatography (GC) are limited due to the chemical complexity of urine, which poorly detects co-eluting low-abundance analytes. Metabolite detection and identification can be improved by applying comprehensive two-dimensional GC, allowing for the discovery of additional viable biomarkers of disease. In this work, we applied comprehensive two-dimensional GC coupled with time-of-flight mass spectrometry (GC × GC-TOFMS) to the analysis of urine samples collected daily across 28-days from 10 healthy female subjects for a personalized approach to female reproductive health monitoring. Through this analysis, we identified 935 unique volatile metabolites. Two statistical methods, a modified T-statistic and Wilcoxon Rank Sum, were applied to analyze differences in metabolome abundance on ovulation days as compared to non-ovulation days. Four metabolites (2-pentanone, 3-penten-2-one, carbon disulfide, acetone) were identified as statistically significant by the modified T-statistic but not the Rank Sum, after a false-discovery rate of 0.1 was set using a Benjamini-Hochberg correction. Subsequent analyses by boxplot indicated that the putative volatile metabolic biomarkers for fertility are expressed in increased or decreased abundance in urine on the day of ovulation. Individual analysis of metabolome expression across 28-days revealed some subject-specific features, which suggest a potential for long-term, personalized fertility monitoring using metabolomics.


Assuntos
Cromatografia Gasosa-Espectrometria de Massas/métodos , Ciclo Menstrual/metabolismo , Metaboloma/fisiologia , Metabolômica/métodos , Acetona/urina , Adolescente , Adulto , Biomarcadores/urina , Dissulfeto de Carbono/urina , Feminino , Humanos , Ciclo Menstrual/urina , Ovulação/metabolismo , Pentanonas/urina , Adulto Jovem
5.
J Chromatogr B Analyt Technol Biomed Life Sci ; 1118-1119: 157-163, 2019 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-31054449

RESUMO

To improve early renal allograft function, it is important to develop a noninvasive diagnostic method for acute T cell-mediated rejection (TCMR). This study aims to explore potential noninvasive urinary biomarkers to screen for acute TCMR in kidney transplant recipients (KTRs) using untargeted metabolomic profiling. Urinary metabolites, collected from KTRs with stable graft function (STA) or acute TCMR episodes, were analyzed using liquid chromatography-mass spectrometry (LC-MS). Multivariate statistical analyses were performed to discriminate differences in urinary metabolites between the two groups. Receiver operating characteristic (ROC) curve analysis was used to evaluate the diagnostic performance of potential urinary biomarkers. Statistical analysis revealed the differences in urinary metabolites between the two groups and indicated several statistically significant metabolic features suitable for potential biomarkers. By comparing the retention times and mass fragmentation patterns of the chemicals in metabolite databases, samples, and standards, six of these features were clearly identified. ROC curve analysis showed the best performance of the training set (area under the curve value, 0.926; sensitivity, 90.0%; specificity, 84.6%) using a panel of five potential biomarkers: guanidoacetic acid, methylimidazoleacetic acid, dopamine, 4-guanidinobutyric acid, and L-tryptophan. The diagnostic accuracy of this model was 62.5% for an independent test dataset. LC-MS-based untargeted metabolomic profiling is a promising method to discriminate between acute TCMR and STA groups. Our model, based on a panel of five potential biomarkers, needs to be further validated in larger scale studies.


Assuntos
Rejeição de Enxerto/diagnóstico , Rejeição de Enxerto/urina , Transplante de Rim , Metaboloma/fisiologia , Metabolômica/métodos , Adulto , Biomarcadores/urina , Cromatografia Líquida de Alta Pressão , Feminino , Rejeição de Enxerto/metabolismo , Humanos , Masculino , Pessoa de Meia-Idade , Reprodutibilidade dos Testes , Linfócitos T/metabolismo , Espectrometria de Massas em Tandem
6.
Plant Physiol Biochem ; 139: 697-706, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31054472

RESUMO

Phosphorus (P) is a major constituent of biomolecules in plant cells, and is an essential plant macronutrient. Low phosphate (Pi) availability in soils is a major constraint on plant growth. Although a complex variety of plant responses to Pi starvation has been well documented, few studies have integrated both global transcriptome and metabolome analyses to shed light on molecular mechanisms underlying metabolic responses to P deficiency. This study is the first time to investigate global profiles of metabolites and transcripts in soybean (Glycine max) roots subjected to Pi starvation through targeted liquid chromatography electrospray ionization mass spectrometry (LC-ESI-MS/MS) and RNA-sequencing analyses. This integrated analysis allows for assessing coordinated transcriptomic and metabolic responses in terms of both pathway enzyme expression and regulatory levels. Between two Pi availability treatments, a total of 155 metabolites differentially accumulated in soybean roots, of which were phosphorylated metabolites, flavonoids and amino acids. Meanwhile, a total of 1644 differentially expressed genes (DEGs) were identified in soybean roots, including 1199 up-regulated and 445 down-regulated genes. Integration of metabolome and transcriptome analyses revealed Pi starvation responsive connection between specific metabolic processes in soybean roots, especially metabolic processes involving phosphorylated metabolites (e.g., phosphorylated lipids and nucleic acids). Taken together, this study suggests that complex molecular responses scavenging internal Pi from phosphorylated metabolites are typical adaptive strategies soybean roots employ as responses to Pi starvation. Identified DEGs will provide potential target region for future efforts to develop P-efficient soybean cultivars.


Assuntos
Metaboloma/fisiologia , Fósforo/deficiência , Raízes de Plantas/genética , Raízes de Plantas/metabolismo , Soja/genética , Soja/metabolismo , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Metaboloma/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Transcriptoma/genética
7.
Artigo em Inglês | MEDLINE | ID: mdl-31071580

RESUMO

Lipid oxidation is one of the most important processes occurring in living cells and has been investigated through stable end-products. Currently, new insights into many physiological and pathophysiological processes provide a measurement of the first products of oxidation, e.g., oxidized glycerophosphatidylcholines (oxGPCs). Here, we evaluate the capacity of untargeted global metabolomics to measure oxGPCs in serum samples. This evaluation covered analytical reproducibility and data quality as well as the ability to capture metabolic alterations in diverse conditions. The analytical evaluation was performed based on the quality control samples, while the comparative analysis was based on the model of the development of type 2 diabetes mellitus (T2DM). The novelty of this approach arises not only from the measurement of oxGPCs instead of lipid peroxide-derived aldehydes but also from the stratification of the patients according to body mass index (BMI). Such a scenario was dictated by the fact that, despite the well-known relationship between obesity and T2DM development, there are lean individuals suffering from T2DM as well as obese people with normal glucose homeostasis. Our results provided evidence to support the ability of nontargeted metabolomics to measure oxGPCs. Comparative analysis of measured oxGPCs revealed differences in the level of oxGPCs either between different stages of disease development (insulin resistance, prediabetes) or BMI groups (normal weight, overweight, obese). The obtained results provided new insights into the metabolic processes leading to the development of T2DM and opened new paths in the investigation of the impact of body mass in T2DM progress.


Assuntos
Diabetes Mellitus Tipo 2/sangue , Glicerilfosforilcolina/sangue , Glicerilfosforilcolina/química , Metaboloma/fisiologia , Metabolômica/métodos , Adulto , Cromatografia Líquida , Diabetes Mellitus Tipo 2/metabolismo , Humanos , Pessoa de Meia-Idade , Oxirredução , Espectrometria de Massas em Tandem
8.
Artigo em Inglês | MEDLINE | ID: mdl-31071582

RESUMO

The emerging requests for handling complex samples in system biology studies highlighted the need to expand the metabolite coverage in metabolomics analysis and to take advantage of the quantitative and targeted assays. Here, we developed a novel workflow to integrate time staggered or mass staggered scan methods with globally optimized targeted-mass spectrometry (GOT-MS), to enable broad metabolites coverage with better stability, repeatability, and quantitative capability. To establish these methods, two scheduled selected reaction monitoring (SRM) methods, time staggered and mass staggered approaches, were configured to achieve optimal sensitivity and scan speed and were combined with the GOT-MS strategy. Both methods took advantage of the systematic selection and rearrangement of all detectable metabolic peaks from a GOT-MS peak list, based on either retention time or m/z of the precursor ions. The established methods were then applied to the metabolic profile-based differentiation of Staphylococcus aureus N315 and N315 ex, an isogenic pair of Methicillin-resistant and susceptible S. aureus (MRSA and MSSA). A total of 464 metabolite peaks was detected successfully from pooled MSSA and MRSA bacterial metabolite extract using the GOT-MS method, and ts/ms-GOT-MS demonstrated better sensitivity and repeatability than the GOT-MS and previously established targeted metabolomics method. The semi-quantitative analysis in a broader metabolome coverage was also achieved with ts/ms-GOT-MS methods. Multivariate statistical analyses were also performed to determine whether metabolic profiling approach could differentiate MSSA from MRSA. The comparison of these methods to GOT-MS and targeted metabolic profiling demonstrated that ts/ms-GOT-MS are significantly improved hybrid metabolomics methods and can be used as promising tools for future studies.


Assuntos
Espectrometria de Massas/métodos , Metaboloma/fisiologia , Metabolômica/métodos , Tipagem Molecular/métodos , Staphylococcus aureus , Biomarcadores/análise , Biomarcadores/metabolismo , Cromatografia Líquida , Modelos Lineares , Staphylococcus aureus Resistente à Meticilina/química , Staphylococcus aureus Resistente à Meticilina/classificação , Staphylococcus aureus Resistente à Meticilina/metabolismo , Reprodutibilidade dos Testes , Staphylococcus aureus/química , Staphylococcus aureus/classificação , Staphylococcus aureus/metabolismo
9.
Gut ; 68(8): 1430-1438, 2019 08.
Artigo em Inglês | MEDLINE | ID: mdl-30971437

RESUMO

OBJECTIVE: To investigate the underlying mechanisms behind changes in glucose homeostasis with delivery of propionate to the human colon by comprehensive and coordinated analysis of gut bacterial composition, plasma metabolome and immune responses. DESIGN: Twelve non-diabetic adults with overweight and obesity received 20 g/day of inulin-propionate ester (IPE), designed to selectively deliver propionate to the colon, a high-fermentable fibre control (inulin) and a low-fermentable fibre control (cellulose) in a randomised, double-blind, placebo-controlled, cross-over design. Outcome measurements of metabolic responses, inflammatory markers and gut bacterial composition were analysed at the end of each 42-day supplementation period. RESULTS: Both IPE and inulin supplementation improved insulin resistance compared with cellulose supplementation, measured by homeostatic model assessment 2 (mean±SEM 1.23±0.17 IPE vs 1.59±0.17 cellulose, p=0.001; 1.17±0.15 inulin vs 1.59±0.17 cellulose, p=0.009), with no differences between IPE and inulin (p=0.272). Fasting insulin was only associated positively with plasma tyrosine and negatively with plasma glycine following inulin supplementation. IPE supplementation decreased proinflammatory interleukin-8 levels compared with cellulose, while inulin had no impact on the systemic inflammatory markers studied. Inulin promoted changes in gut bacterial populations at the class level (increased Actinobacteria and decreased Clostridia) and order level (decreased Clostridiales) compared with cellulose, with small differences at the species level observed between IPE and cellulose. CONCLUSION: These data demonstrate a distinctive physiological impact of raising colonic propionate delivery in humans, as improvements in insulin sensitivity promoted by IPE and inulin were accompanied with different effects on the plasma metabolome, gut bacterial populations and markers of systemic inflammation.


Assuntos
Microbioma Gastrointestinal/fisiologia , Insulina/metabolismo , Inulina , Metaboloma/fisiologia , Obesidade , Sobrepeso , Adulto , Índice de Massa Corporal , Estudos Cross-Over , Suplementos Nutricionais , Método Duplo-Cego , Fezes/microbiologia , Feminino , Humanos , Inflamação/metabolismo , Resistência à Insulina/fisiologia , Inulina/administração & dosagem , Inulina/metabolismo , Masculino , Pessoa de Meia-Idade , Obesidade/diagnóstico , Obesidade/dietoterapia , Obesidade/metabolismo , Sobrepeso/diagnóstico , Sobrepeso/dietoterapia , Sobrepeso/metabolismo , Propionatos/administração & dosagem , Propionatos/metabolismo , Resultado do Tratamento
10.
Environ Sci Pollut Res Int ; 26(17): 17362-17372, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31012076

RESUMO

Epoxiconazole is a broad-spectrum fungicide described as highly persistent in soil and as such can be considered as an abiotic agent like other problematic agrochemicals. Furthermore, the plant phenotyping tool involving non-invasive monitoring of plant-emitted volatile organic compounds (VOCs) may be useful in the identification of metabolic markers for abiotic stress. We therefore decided to profile the VOCs from secondary metabolism of oilseed rape through a dose-response experiment under several epoxiconazole concentrations (0, 0.01, 0.1 and 1 mg L-1). VOC collections of 35-day-old whole plantlets were performed through a dynamic headspace sampling technique under defined and controlled conditions. The plantlets grew freely within a home-made, laboratory and high-throughput glass chamber without any disturbance. Putative metabolic markers were analysed using a targeted metabolomic approach based on TD-GC-MS method coupled with data acquisition in SIM mode in order to focus on terpenes and sulphur-containing volatiles. Chromatograms of emitted terpenes were achieved accurately for the 35-day-old oilseed rape plantlets. We also analysed the presence of sulphur-containing volatiles in samples of shoot and root tissues using an innovative DHS-TD-GC-MS method, but no difference was found between qualitative profiles. Nevertheless, we demonstrated through this experiment that sesquiterpenes such as ß-elemene and (E,E)-α-farnesene are involved in epoxiconazole dose-response. In particular, (E,E)-α-farnesene could serve as a metabolic marker of fungicide exposure for oilseed rape plantlets.


Assuntos
Brassica napus/fisiologia , Compostos de Epóxi/toxicidade , Poluentes do Solo/toxicidade , Terpenos/metabolismo , Triazóis/toxicidade , Brassica napus/efeitos dos fármacos , Brassica napus/metabolismo , Cromatografia Gasosa-Espectrometria de Massas , Metaboloma/efeitos dos fármacos , Metaboloma/fisiologia , Metabolômica , Extratos Vegetais/metabolismo , Sesquiterpenos , Estresse Fisiológico , Testes de Toxicidade , Compostos Orgânicos Voláteis/análise
11.
Int J Mol Sci ; 20(9)2019 Apr 26.
Artigo em Inglês | MEDLINE | ID: mdl-31027387

RESUMO

Plant regeneration via somatic embryogenesis (SE) is a key step during genetic engineering. In the current study, integrated widely targeted metabolomics and RNA sequencing were performed to investigate the dynamic metabolic and transcriptional profiling of cotton SE. Our data revealed that a total of 581 metabolites were present in nonembryogenic staged calli (NEC), primary embryogenic calli (PEC), and initiation staged globular embryos (GE). Of the differentially accumulated metabolites (DAMs), nucleotides, and lipids were specifically accumulated during embryogenic differentiation, whereas flavones and hydroxycinnamoyl derivatives were accumulated during somatic embryo development. Additionally, metabolites related to purine metabolism were significantly enriched in PEC vs. NEC, whereas in GE vs. PEC, DAMs were remarkably associated with flavonoid biosynthesis. An association analysis of the metabolome and transcriptome data indicated that purine metabolism and flavonoid biosynthesis were co-mapped based on the Kyoto encyclopedia of genes and genomes (KEGG) database. Moreover, purine metabolism-related genes associated with signal recognition, transcription, stress, and lipid binding were significantly upregulated. Moreover, several classic somatic embryogenesis (SE) genes were highly correlated with their corresponding metabolites that were involved in purine metabolism and flavonoid biosynthesis. The current study identified a series of potential metabolites and corresponding genes responsible for SE transdifferentiation, which provides a valuable foundation for a deeper understanding of the regulatory mechanisms underlying cell totipotency at the molecular and biochemical levels.


Assuntos
Flavonoides/metabolismo , Gossypium/metabolismo , Purinas/metabolismo , Transdiferenciação Celular , Regulação da Expressão Gênica de Plantas/genética , Gossypium/genética , Metaboloma/genética , Metaboloma/fisiologia , Transcriptoma/genética
12.
J Pharm Biomed Anal ; 170: 305-320, 2019 Jun 05.
Artigo em Inglês | MEDLINE | ID: mdl-30954021

RESUMO

Achyranthes bidentate (AB) is a typical traditional Chinese medicine (TCM) that has been widely used in clinical practices for more than a thousand years. Modern pharmacological studies have shown that triterpene saponins are the main pharmacological active ingredients in AB. Meanwhile, the poor oral bioavailability of triterpene saponins in AB indicates that these ingredients are probably metabolized by intestinal microflora before absorption. In this work, an integrated analysis based on ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS) combined with a metabolism platform was developed to identify the chemical constituents and intestinal metabolic profiles of triterpene saponins in AB. As a result, a total of 40 triterpene saponins (including thirty-eight oleanane-type, one hederagenin-type and one machaerinate-type triterpene saponin) were identified from the AB extract. Moreover, 39 biotransformation products mediated by intestinal microflora were characterized, which mainly underwent four metabolic reactions including deglycosylation, glycosylation, oxidation and dehydrogenation. To our knowledge, the in vitro metabolites of AB through intestinal microflora metabolism, especially triterpene saponins, have not been studied previously. The obtained results could be helpful for the further evaluation of the pharmacokinetics and the pharmacological activity of triterpene saponins of AB in vivo.


Assuntos
Achyranthes/metabolismo , Biotransformação/fisiologia , Microbioma Gastrointestinal/fisiologia , Intestinos/microbiologia , Metaboloma/fisiologia , Saponinas/metabolismo , Cromatografia Líquida de Alta Pressão/métodos , Medicamentos de Ervas Chinesas/metabolismo , Medicina Tradicional Chinesa/métodos , Espectrometria de Massas em Tandem/métodos , Triterpenos/metabolismo
13.
Artigo em Inglês | MEDLINE | ID: mdl-30951967

RESUMO

The prevalence of chronic kidney disease (CKD) is increasing worldwide. New technical approaches are needed to improve early diagnosis, disease understanding and patient monitoring, and to evaluate new therapies. Metabolomics, as a prime candidate in the field of CKD research, aims to comprehensively analyze the metabolic complexity of biological systems. An extensive analysis of the metabolites contained in biofluids is therefore needed, and the combination of data obtained from multiple analytical platforms constitutes a promising methodological approach. This study presents an original workflow based on complementary chromatographic conditions, reversed-phase and hydrophilic interaction chromatography hyphenated to mass spectrometry to improve the polar metabolome coverage coupled with a univocal metabolite annotation strategy enabling a rapid access to the biological interpretation. This multiplatform workflow was applied in a CKD cohort study to assess plasma metabolic profile modifications related to renal disease. Multivariate analysis of 278 endogenous annotated metabolites enabled patient stratification with respect to CKD stages and helped to generate new biological insights, while also confirming the relevance of tryptophan metabolism pathway in this condition.


Assuntos
Cromatografia Líquida/métodos , Espectrometria de Massas/métodos , Metabolômica/métodos , Insuficiência Renal Crônica/sangue , Insuficiência Renal Crônica/diagnóstico , Adulto , Biomarcadores/sangue , Estudos de Casos e Controles , Humanos , Metaboloma/fisiologia , Reprodutibilidade dos Testes
14.
Artigo em Inglês | MEDLINE | ID: mdl-30991202

RESUMO

Necrotizing enterocolitis (NEC) is a leading cause of gastrointestinal morbidity and mortality in preterm neonates. The aim of this pilot study was to explore using metabolomics alternations in the urine metabolites related to NEC that could possibly serve as diagnostic biomarkers of the disease. Urine samples were prospectively collected at the day of initial evaluation for NEC from 15 diseased preterm neonates (five Bell's stage I and ten stage II/III) and an equal number of matched controls. Urine metabolic profiles were assessed using non-targeted nuclear magnetic resonance spectroscopy and targeted liquid chromatography-tandem mass spectrometry monitoring 108 metabolites. Multivariate statistical models with data from either analytical approach showed clear separation between the metabolic profiles of neonates with NEC and controls. Twenty-five discriminant metabolites were identified belonging to amino and organic acids, sugars and vitamins. A number of metabolite combinations were found to have an excellent diagnostic performance in detecting neonates developing NEC. Our results show that the metabolic profile of neonates with NEC differs significantly from that of controls, making possible their separation using urine metabolomic analysis. Nevertheless, whether the small set of significant metabolites detected in this investigation could be used as early diagnostic biomarkers of NEC should be validated in larger studies.


Assuntos
Enterocolite Necrosante/diagnóstico , Enterocolite Necrosante/urina , Metaboloma/fisiologia , Metabolômica/métodos , Biomarcadores/urina , Estudos de Casos e Controles , Cromatografia Líquida/métodos , Feminino , Humanos , Recém-Nascido , Masculino , Projetos Piloto , Espectrometria de Massas em Tandem/métodos
15.
Artigo em Inglês | MEDLINE | ID: mdl-30999273

RESUMO

Changes in metabolites composition can reflect currently present pathological processes in a living organism and constitute a basis for diagnosis and treatment improvements. Thus, the multiplatform metabolomics approach was applied for the investigation of molecular mechanisms of chronic kidney disease (CKD) progression. The high-performance liquid chromatography coupled with time-of-flight mass spectrometry (HPLC-TOF-MS) and gas chromatography coupled with triple quadrupole mass spectrometry (GC-QqQ/MS) serum metabolic fingerprinting followed by uni- and multivariate statistical analysis was carried out to determine metabolic pattern differentiating CKD patients and healthy controls. Furthermore, metabolites changes between stage 3 and 4 of the disease, as well as health status were investigated. The progression of the disease was found to be related to alterations in acylcarnitine, amino acid, lysophospholipid and carbohydrate metabolism. Elevated levels of serum acylcarnitines, sugar alcohols, and organic acids, as well as decreased levels of lysophospholipids, and amino acids, were found to be statistically significant for CKD progression. The obtained results confirm the utility of metabolomics approach as a tool for an explanation of molecular processes underlying CKD development.


Assuntos
Metaboloma/fisiologia , Metabolômica/métodos , Insuficiência Renal Crônica , Adulto , Idoso , Biomarcadores/sangue , Biomarcadores/metabolismo , Carnitina/análogos & derivados , Carnitina/sangue , Estudos de Casos e Controles , Cromatografia Líquida de Alta Pressão/métodos , Análise por Conglomerados , Progressão da Doença , Feminino , Cromatografia Gasosa-Espectrometria de Massas/métodos , Humanos , Masculino , Espectrometria de Massas , Pessoa de Meia-Idade , Insuficiência Renal Crônica/sangue , Insuficiência Renal Crônica/diagnóstico , Insuficiência Renal Crônica/metabolismo
16.
Artigo em Inglês | MEDLINE | ID: mdl-31009899

RESUMO

Liquid chromatography (LC) hyphenated to mass spectrometry is currently the most widely used means of determining metabolic phenotypes via both untargeted and targeted analysis. At present a range of analytical separations, including reversed-phase, hydrophilic interaction and ion-pair LC are employed to maximise metabolome coverage with ultra (high) performance liquid chromatography (UHPLC) increasingly displacing conventional high performance liquid chromatography because of the need for short analysis times and high peak capacity in such applications. However, it is widely recognized that these methodologies do not entirely solve the problems facing researchers trying to perform comprehensive metabolic phenotyping and in addition to these "routine" approaches there are continuing investigations of alternative separation methods including 2-dimensional/multi column approaches. These involve either new stationary phases or multidimensional combinations of the more conventional materials currently used, as well as application of miniaturization or "new" approaches such as supercritical HP and UHP- chromatographic separations. There is also a considerable amount of interest in the combination of chromatographic and ion mobility separations, with the latter providing both an increase in resolution and the potential to provide additional structural information via the determination of molecular collision cross section data. However, key problems remain to be solved including ensuring quality, comparability across different laboratories and the ever present difficulty of identifying unknowns.


Assuntos
Cromatografia Líquida , Espectrometria de Massas , Metabolômica , Animais , Biomarcadores/sangue , Biomarcadores/urina , Humanos , Metaboloma/fisiologia
17.
J Chromatogr B Analyt Technol Biomed Life Sci ; 1118-1119: 25-32, 2019 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-31005771

RESUMO

Polycystic kidney disease (PKD) encompasses a spectrum of inherited disorders that lead to end-stage renal disease (ESRD). There is no cure for PKD and current treatment options are limited to renal replacement therapy and transplantation. A better understanding of the pathobiology of PKD is needed for the development of new, less invasive treatments. The Lewis Polycystic Kidney (LPK) rat phenotype has been characterized and classified as a model of nephronophthisis (NPHP9, caused by mutation of the Nek8 gene) for which polycystic kidneys are one of the main pathologic features. The aim of this study was to use a GC-MS-based untargeted metabolomics approach to determine key biochemical changes in kidney and liver tissue of the LPK rat. Tissues from 16-week old LPK (n = 10) and Lewis age- and sex-matched control animals (n = 11) were used. Principal component analysis (PCA) distinguished signal corrected metabolite profiles from Lewis and LPK rats for kidney (PC-1 77%) and liver (PC-1 46%) tissue. There were marked differences in the metabolite profiles of the kidney tissues with 122 deconvoluted features significantly different between the LPK and Lewis strains. The metabolite profiles were less marked between strains for liver samples with 30 features significantly different. Five biochemical pathways showed three or more significantly altered metabolites: transcription/translation, arginine and proline metabolism, alpha-linolenic and linoleic acid metabolism, the citric acid cycle, and the urea cycle. The results of this study validate and complement the current literature and are consistent with the understood pathobiology of PKD.


Assuntos
Cromatografia Gasosa-Espectrometria de Massas/métodos , Rim/metabolismo , Fígado/metabolismo , Metabolômica/métodos , Doenças Renais Policísticas/metabolismo , Aminoácidos/análise , Aminoácidos/metabolismo , Animais , Biomarcadores/análise , Biomarcadores/metabolismo , Feminino , Masculino , Metaboloma/fisiologia , Ratos , Reprodutibilidade dos Testes
18.
Psychopharmacology (Berl) ; 236(5): 1531-1544, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30903211

RESUMO

RATIONALE: Increasing evidence has demonstrated that changes in the gut microbiome, including those associated with dietary influences, are associated with alterations in many physiological processes. Alcohol consumption is common across human cultures and is likely to have a major effect on the gut microbiome, but there remains a paucity of information on its effects in primates. OBJECTIVES: The effects of chronic alcohol consumption on the primate gut microbiome and metabolome were studied in rhesus macaques that were freely drinking alcohol. The objectives of the study were to determine what changes occurred in the gut microbiome following long-term exposure to alcohol and if these changes were reversible following a period of abstinence. METHODS: Animals consuming alcohol were compared to age-matched controls without access to alcohol and were studied before and after a period of abstinence. Fecal samples from rhesus macaques were used for 16S rRNA sequencing to profile the gut microbiome and for metabolomic profiling using mass spectrometry. RESULTS: Alcohol consumption resulted in a loss of alpha-diversity in rhesus macaques, though this was partially ameliorated by a period of abstinence. Higher levels of Firmicutes were observed in alcohol-drinking animals at the expense of a number of other microbial taxa, again normalizing in part with a period of abstinence. Metabolomic changes were primarily associated with differences in glycolysis when animals were consuming alcohol and differences in fatty acids when alcohol-drinking animals became abstinent. CONCLUSIONS: The consumption of alcohol has specific effects on the microbiome and metabolome of rhesus macaques independent of secondary influences. Many of these changes are reversed by a relatively short period of abstinence.


Assuntos
Consumo de Bebidas Alcoólicas/efeitos adversos , Etanol/toxicidade , Microbioma Gastrointestinal/efeitos dos fármacos , Metaboloma/efeitos dos fármacos , Consumo de Bebidas Alcoólicas/genética , Consumo de Bebidas Alcoólicas/metabolismo , Animais , Etanol/administração & dosagem , Fezes/microbiologia , Microbioma Gastrointestinal/fisiologia , Humanos , Macaca mulatta , Masculino , Metaboloma/fisiologia , RNA Ribossômico 16S/genética , RNA Ribossômico 16S/metabolismo
19.
Biomed Chromatogr ; 33(7): e4533, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-30891765

RESUMO

To investigate the characteristic of neuromyelitis optica spectrum disorder (NMOSD) in plasma and cerebrospinal fluid, UHPLC-MS was used to identify metabolites by metabolite extraction and on-machine detection. Multivariate analysis methods were used to complete differential metabolite screening and metabolic pathway analysis. The content of eight substances, such as tryptophan and l-glutamic acid in the plasma of NMOSD patients, was higher than that of the healthy control group. Moreover, no differential metabolite was found in the plasma of patients with AQP-4 antibody-positive and antibody-negative NMOSD. The content of five substances including 3-hydroxybutyric acid in cerebrospinal fluid of patients with NMOSD was reduced. We demonstrated that the distribution of metabolites in plasma between NMOSD patients and healthy counterparts was significantly different. However, there is no significant difference in plasma metabolites between AQP-4 antibody positive and negative NMOSD. There were some differences in metabolites between the cerebrospinal fluid of NMOSD patients and that of healthy controls. A variety of amino-acid abnormalities, sphingomyelin dysfunction, energy metabolism and mitochondrial dysfunction are involved in the pathogenesis of NMOSD.


Assuntos
Metaboloma/fisiologia , Metabolômica/métodos , Neuromielite Óptica/sangue , Neuromielite Óptica/metabolismo , Biomarcadores/sangue , Biomarcadores/metabolismo , Cromatografia Líquida de Alta Pressão , Estudos de Coortes , Humanos , Análise dos Mínimos Quadrados , Espectrometria de Massas
20.
Phytother Res ; 33(5): 1501-1509, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30895694

RESUMO

Nonalcoholic fatty liver disease (NAFLD) is the most common form of chronic liver diseases associated with unfavorable metabolic profiles and oxidative stress parameters. This study was designed to determine the effects of purslane seeds consumption with a low-calorie diet on insulin resistance, lipid profile, and oxidative stress indices in patients with NAFLD. This randomized controlled clinical trial was conducted on 54 individuals with NAFLD. Subjects were randomly assigned to consume either 10 g/day of purslane seeds sachet before breakfast and dinner in addition to a low-calorie diet (n = 27) or only the low-calorie diet (n = 27) for 8 weeks. Fasting blood samples were collected at the beginning and end of the study to measure relevant variables. Intake of purslane seeds with the low-calorie diet led to a significant decrease in serum concentrations of fasting blood sugar (FBS; -3.52 ± 10.45 compared with 3.03 ± 9.01 mg/dl, P = 0.017), quantitative insulin sensitivity check index (QUICKI; 0.13 ± 0.27 compared with -0.002 ± 0.016, P = 0.017), total cholesterol (4.33 ± 34.04 compared with 23.48 ± 29.47 mg/dl, P = 0.032), and low-density lipoprotein cholesterol (LDL-C; -4.35 ± 22.65 compared with 11.82 ± 16.08 mg/dl, P = 0.004) after intervention. Compared with the control group, purslane seeds consumption with adherence to a low-calorie diet had beneficial effects on FBS, HOMA-IR, QUICKI, serum total, and LDL-C in patients with NAFLD but did not affect other glycemic, lipid profile, and oxidative stress parameters.


Assuntos
Metaboloma/fisiologia , Hepatopatia Gordurosa não Alcoólica/dietoterapia , Plantas/química , Portulaca/química , Sementes/química , Adulto , Feminino , Humanos , Masculino , Hepatopatia Gordurosa não Alcoólica/patologia
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