RESUMO
Prostate cancer (PCa) is the most common cancer form in males in many European and American countries, but there are still open questions regarding its etiology. Untargeted metabolomics can produce an unbiased global metabolic profile, with the opportunity for uncovering new plasma metabolites prospectively associated with risk of PCa, providing insights into disease etiology. We conducted a prospective untargeted liquid chromatography-mass spectrometry (LC-MS) metabolomics analysis using prediagnostic fasting plasma samples from 752 PCa case-control pairs nested within the Northern Sweden Health and Disease Study (NSHDS). The pairs were matched by age, BMI, and sample storage time. Discriminating features were identified by a combination of orthogonal projection to latent structures-effect projections (OPLS-EP) and Wilcoxon signed-rank tests. Their prospective associations with PCa risk were investigated by conditional logistic regression. Subgroup analyses based on stratification by disease aggressiveness and baseline age were also conducted. Various free fatty acids and phospholipids were positively associated with overall risk of PCa and in various stratification subgroups. Aromatic amino acids were positively associated with overall risk of PCa. Uric acid was positively, and glucose negatively, associated with risk of PCa in the older subgroup. This is the largest untargeted LC-MS based metabolomics study to date on plasma metabolites prospectively associated with risk of developing PCa. Different subgroups of disease aggressiveness and baseline age showed different associations with metabolites. The findings suggest that shifts in plasma concentrations of metabolites in lipid, aromatic amino acid, and glucose metabolism are associated with risk of developing PCa during the following two decades.
Assuntos
Ácidos Graxos não Esterificados , Neoplasias da Próstata , Masculino , Humanos , Estudos de Casos e Controles , Ácido Úrico , Suécia/epidemiologia , Metabolômica/métodos , Espectrometria de Massas , Neoplasias da Próstata/diagnóstico , Neoplasias da Próstata/epidemiologia , Aminoácidos Aromáticos , GlucoseRESUMO
Androgen deprivation therapy (ADT) remains a key approach in the treatment of prostate cancer (PCa). However, PCa inevitably relapses and becomes ADT resistant. Besides androgens, there is evidence that thyroid hormone thyroxine (T4) and its active form 3,5,3'-triiodo-L-thyronine (T3) are involved in the progression of PCa. Epidemiologic evidences show a higher incidence of PCa in men with elevated thyroid hormone levels. The thyroid hormone binding protein µ-Crystallin (CRYM) mediates intracellular thyroid hormone action by sequestering T3 and blocks its binding to cognate receptors (TRα/TRß) in target tissues. We show in our study that low CRYM expression levels in PCa patients are associated with early biochemical recurrence and poor prognosis. Moreover, we found a disease stage-specific expression of CRYM in PCa. CRYM counteracted thyroid and androgen signaling and blocked intracellular choline uptake. CRYM inversely correlated with [18F]fluoromethylcholine (FMC) levels in positron emission tomography/magnetic resonance imaging of PCa patients. Our data suggest CRYM as a novel antagonist of T3- and androgen-mediated signaling in PCa. The role of CRYM could therefore be an essential control mechanism for the prevention of aggressive PCa growth.
Assuntos
Cristalinas/genética , Cristalinas/metabolismo , Regulação para Baixo , Neoplasias da Próstata/patologia , Transdução de Sinais , Linhagem Celular Tumoral , Colina/administração & dosagem , Colina/análogos & derivados , Estudos de Coortes , Perfilação da Expressão Gênica , Regulação Neoplásica da Expressão Gênica , Humanos , Masculino , Metabolômica , Estadiamento de Neoplasias , Células PC-3 , Tomografia por Emissão de Pósitrons combinada à Tomografia Computadorizada , Prognóstico , Neoplasias da Próstata/diagnóstico por imagem , Neoplasias da Próstata/genética , Neoplasias da Próstata/metabolismo , Receptores Androgênicos/genética , Receptores Androgênicos/metabolismo , Receptores dos Hormônios Tireóideos/genética , Análise de Sequência de RNA , Análise Serial de Tecidos , Tri-Iodotironina/antagonistas & inibidores , Tri-Iodotironina/metabolismo , Cristalinas muRESUMO
We have recently presented an Automated Quantification Algorithm (AQuA) and demonstrated its utility for rapid and accurate absolute metabolite quantification in 1H NMR spectra in which positions and line widths of signals were predicted from a constant metabolite spectral library. The AQuA quantifies based on one preselected signal per metabolite and employs library spectra to model interferences from other metabolite signals. However, for some types of spectra, the interspectral deviations of signal positions and line widths can be pronounced; hence, interferences cannot be modeled using a constant spectral library. We here address this issue and present an improved AQuA that handles interspectral deviations. The improved AQuA monitors and characterizes the appearance of specific signals in each spectrum and automatically adjusts the spectral library to model interferences accordingly. The performance of the improved AQuA was tested on a large data set from plasma samples collected using ethylenediaminetetraacetic acid (EDTA) as an anticoagulant (n = 772). These spectra provided a suitable test system for the improved AQuA since EDTA signals (i) vary in intensity, position, and line width between spectra and (ii) interfere with many signals from plasma metabolites targeted for quantification (n = 54). Without the improvement, ca. 20 out of the 54 metabolites would have been overestimated. This included acetylcarnitine and ornithine, which are considered particularly difficult to quantify with 1H NMR in EDTA-containing plasma. Furthermore, the improved AQuA performed rapidly (<10 s for all spectra). We believe that the improved AQuA provides a basis for automated quantification in other data sets where specific signals show interspectral deviations.
Assuntos
Algoritmos , Metabolômica , Ácido Edético , Imageamento por Ressonância Magnética , Espectroscopia de Ressonância MagnéticaRESUMO
INTRODUCTION: Hyperinsulinaemia and insulin resistance (IR) are strongly associated with obesity and are forerunners of type 2 diabetes. Little is known about metabolic alterations separately associated with obesity, hyperinsulinaemia/IR and impaired glucose tolerance (IGT) in adolescents. OBJECTIVES: To identify metabolic alterations associated with obesity, hyperinsulinaemia/IR and hyperinsulinaemia/IR combined with IGT in obese adolescents. METHODS: 81 adolescents were stratified into four groups based on body mass index (lean vs. obese), insulin responses (normal insulin (NI) vs. high insulin (HI)) and glucose responses (normal glucose tolerance (NGT) vs. IGT) after an oral glucose tolerance test (OGTT). The groups comprised: (1) healthy lean with NI and NGT, (2) obese with NI and NGT, (3) obese with HI and NGT, and (4) obese with HI and IGT. Targeted nuclear magnetic resonance-based metabolomics analysis was performed on fasting and seven post-OGTT plasma samples, followed by univariate and multivariate statistical analyses. RESULTS: Two groups of metabolites were identified: (1) Metabolites associated with insulin response level: adolescents with HI (groups 3-4) had higher concentrations of branched-chain amino acids and tyrosine, and lower concentrations of serine, glycine, myo-inositol and dimethylsulfone, than adolescents with NI (groups 1-2). (2) Metabolites associated with obesity status: obese adolescents (groups 2-4) had higher concentrations of acetylcarnitine, alanine, pyruvate and glutamate, and lower concentrations of acetate, than lean adolescents (group 1). CONCLUSIONS: Obesity is associated with shifts in fat and energy metabolism. Hyperinsulinaemia/IR in obese adolescents is also associated with increased branched-chain and aromatic amino acids.
Assuntos
Hiperinsulinismo/metabolismo , Resistência à Insulina , Metaboloma , Metabolômica , Obesidade/metabolismo , Adolescente , Biomarcadores , Criança , Estudos Transversais , Feminino , Humanos , Hiperinsulinismo/sangue , Hiperinsulinismo/epidemiologia , Estudos Longitudinais , Masculino , Metabolômica/métodos , Obesidade/sangue , Obesidade/epidemiologia , Obesidade Infantil/sangue , Obesidade Infantil/metabolismo , Puberdade , Suécia/epidemiologiaRESUMO
BACKGROUND: Prostate cancer is the second most frequently diagnosed cancer in men. Metabolomics can potentially provide new insights into the aetiology of prostate cancer by identifying new metabolic risk factors. This study investigated the prospective association between plasma metabolite concentrations and prostate cancer risk, both overall and by stratifying for disease aggressiveness and baseline age. METHODS: In a case-control study nested in the Northern Sweden Health and Disease Study, pre-diagnostic concentrations of 148 plasma metabolites were determined using targeted mass spectrometry- and nuclear magnetic resonance-based metabolomics in 777 prostate cancer cases (follow-up ≥ 5 years) and 777 matched controls. Associations between prostate cancer risk and metabolite concentrations were investigated using conditional logistic regression conditioned on matching factors (body mass index, age and sample storage time). Corrections for multiple testing were performed using false discovery rate (20%) and Bonferroni. Metabolomics analyses generated new hypotheses, which were investigated by leveraging food frequency questionnaires (FFQs) and oral glucose tolerance tests performed at baseline. RESULTS: After correcting for multiple testing, two lysophosphatidylcholines (LPCs) were positively associated with risk of overall prostate cancer (all ages and in older subjects). The strongest association was for LPC C17:0 in older subjects (OR = 2.08; 95% CI 1.45-2.98; p < 0.0001, significant also after the Bonferroni correction). Observed associations with risk of overall prostate cancer in younger subjects were positive for glycine and inverse for pyruvate. For aggressive prostate cancer, there were positive associations with six glycerophospholipids (LPC C17:0, LPC C20:3, LPC C20:4, PC ae C38:3, PC ae C38:4 and PC ae C40:2), while there was an inverse association with acylcarnitine C18:2. Moreover, plasma LPC C17:0 concentrations positively correlated with estimated dietary intake of fatty acid C17:0 from the FFQs. The associations between glycerophospholipids and prostate cancer were stronger in case-controls with normal glucose tolerance. CONCLUSIONS: Several glycerophospholipids were positively associated with risk of overall and aggressive prostate cancer. The strongest association was observed for LPC C17:0. The associations between glycerophospholipids and prostate cancer risk were stronger in case-controls with normal glucose tolerance, suggesting a link between the glucose metabolism status and risk of prostate cancer.
Assuntos
Espectrometria de Massas/métodos , Metabolômica/métodos , Neoplasias da Próstata/sangue , Adulto , Estudos de Casos e Controles , Seguimentos , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Fatores de Risco , SuéciaRESUMO
PURPOSE: Irritable bowel syndrome is a gastrointestinal disorder which can influence human metabolism. It has been demonstrated that probiotics are beneficial in controlling IBS. Thus, the main objective of the present study was to determine metabolic changes in response to diarrhea predominant irritable bowel syndrome (IBS-D) and to investigate the metabolic effects of a synbiotic intervention on serum, urine, and stool samples from IBS-D patients and healthy controls using proton nuclear magnetic resonance (1HNMR). METHODS: A 1HNMR-based metabolomics study was conducted on urine and serum metabolites from 16 healthy and eight IBS-D participants at baseline and after 4 weeks of a synbiotic yogurt intervention. RESULTS: At the baseline, serum acetoacetate, myo-inositol, and sarcosine concentrations were higher and threonine and methionine concentrations were lower in the IBS-D cohort than the control group. Moreover, Indoxyl-sulfate concentration of urine was lower and dimethylamine and taurine were higher in the IBS-D group. After intervention, serum concentration of ketone bodies decreased, choline, phenylalanine, and branched-chain amino acids increased in IBS-D group. Metabolomics analysis indicated a shift in one-carbon metabolism. Thus, the level of serum homocysteine was determined and found to be higher in the IBS-D cohort at baseline, and then decreased after the intervention. CONCLUSION: IBS causes a shift in one-carbon metabolism and these changes can be reversed by a synbiotic intervention. An increase in the number of fecal Lactobacilli and an improvement in the health status of IBS-D patients were also observed in response to intervention.
Assuntos
Diarreia/metabolismo , Microbioma Gastrointestinal , Síndrome do Intestino Irritável/metabolismo , Metabolômica/métodos , Simbióticos/administração & dosagem , Iogurte , Adulto , Feminino , Trato Gastrointestinal/diagnóstico por imagem , Trato Gastrointestinal/microbiologia , Humanos , Irã (Geográfico) , Síndrome do Intestino Irritável/complicações , Espectroscopia de Ressonância Magnética , MasculinoRESUMO
BACKGROUND: Obesity in dogs is an increasing problem associated with morbidity, shortened life span and poor life quality. Overweight dogs exhibit postprandial hyperlipidaemia, highlighting the need to identify potential dysregulations in lipid metabolism. This study investigated metabolites related to lipid metabolism (i.e. acylcarnitines and taurine) and phospholipids in a feed-challenge test and aimed to identify metabolic variations in spontaneously overweight dogs. Twenty-eight healthy male Labrador Retriever dogs were included, 12 of which were classified as lean (body condition score (BCS) 4-5 on a 9-point scale) and 16 as overweight (BCS 6-8). After overnight fasting (14-17 h), fasting blood samples were collected and dogs were fed a high-fat meal followed by postprandial blood sample collection hourly for 4 h. Liquid chromatography-time of flight mass spectrometry (LC-TOFMS) was used to identify plasma metabolites and phospholipids. Multivariate models, mixed model repeated measures and linear regression analyses were used for data interpretation. RESULTS: In all dogs, propionylcarnitine, stearoylcarnitine and nine phospholipids increased in response to food intake, while vaccenylcarnitine decreased (P ≤ 0.005 for all). Overall, carnitine and acetylcarnitine signal areas in the feed-challenge test were lower in overweight dogs (P ≤ 0.004). Notably, fasting plasma acetylcarnitine was lower in overweight dogs than in lean dogs (P = 0.001) and it did not change in response to feeding. The latter finding was in contrast to the decreased acetylcarnitine signal area found in lean dogs at one hour postprandially (P < 0.0001). One fasting phosphatidylcholine (PCaa C38:4) was higher in prominently overweight dogs (BCS > 6) than in lean dogs (P < 0.05). CONCLUSIONS: Plasma carnitine status was overall lower in spontaneously overweight dogs than in lean dogs in this cohort of healthy Labrador Retriever dogs, indicating a potential carnitine insufficiency in the overweight group. The acetylcarnitine response in overweight dogs indicated decreased fatty acid oxidation at fasting and metabolic inflexibility to food intake. Further studies on metabolic inflexibility and its potential role in the metabolism of overweight dogs are warranted.
Assuntos
Doenças do Cão/metabolismo , Ingestão de Alimentos , Sobrepeso/veterinária , Animais , Carnitina/análogos & derivados , Carnitina/sangue , Doenças do Cão/etiologia , Cães , Ingestão de Alimentos/fisiologia , Metabolismo dos Lipídeos , Masculino , Sobrepeso/etiologia , Sobrepeso/metabolismo , Fosfolipídeos/sangueRESUMO
A key limiting step for high-throughput NMR-based metabolomics is the lack of rapid and accurate tools for absolute quantification of many metabolites. We developed, implemented, and evaluated an algorithm, AQuA (Automated Quantification Algorithm), for targeted metabolite quantification from complex 1H NMR spectra. AQuA operates based on spectral data extracted from a library consisting of one standard calibration spectrum for each metabolite. It uses one preselected NMR signal per metabolite for determining absolute concentrations and does so by effectively accounting for interferences caused by other metabolites. AQuA was implemented and evaluated using experimental NMR spectra from human plasma. The accuracy of AQuA was tested and confirmed in comparison with a manual spectral fitting approach using the ChenomX software, in which 61 out of 67 metabolites quantified in 30 human plasma spectra showed a goodness-of-fit (r2) close to or exceeding 0.9 between the two approaches. In addition, three quality indicators generated by AQuA, namely, occurrence, interference, and positional deviation, were studied. These quality indicators permit evaluation of the results each time the algorithm is operated. The efficiency was tested and confirmed by implementing AQuA for quantification of 67 metabolites in a large data set comprising 1342 experimental spectra from human plasma, in which the whole computation took less than 1 s.
Assuntos
Algoritmos , Análise Química do Sangue/métodos , Sangue/metabolismo , Ensaios de Triagem em Larga Escala/métodos , Metabolômica/métodos , Humanos , Masculino , Espectroscopia de Prótons por Ressonância Magnética/estatística & dados numéricosRESUMO
PURPOSE: The change in serum metabolic response from fasting state to postprandial state provides novel insights into the impact of a single meal on human metabolism. Therefore, this study explored changes in serum metabolite profile after a single meal. METHODS: Nineteen healthy postmenopausal women with normal glucose tolerance participated in the study. They received a meal consisting of refined wheat bread (50 g carbohydrates, 9 g protein, 4.2 g fat and 2.7 g dietary fibre), 40 g cucumber and 300 mL noncaloric orange drink. Blood samples were collected at fasting and five postprandial time points. Metabolic profile was measured by nuclear magnetic resonance and targeted liquid chromatography-mass spectrometry. Changes over time were assessed with multivariate models and ANOVA, with baseline as control. RESULTS: The metabolomic analyses demonstrated alterations in phospholipids, amino acids and their breakdown products, glycolytic products, acylcarnitines and ketone bodies after a single meal. More specifically, phosphatidylcholines, lysophosphatidylcholines and citrate displayed an overall declining pattern, while leucine, isoleucine, methionine and succinate increased initially but declined thereafter. A sharp decline in acylcarnitines and ketone bodies and increase in glycolytic products postprandially suggest a switch in the body's energy source from ß-oxidation to glycolysis. Moreover, individuals with relatively high postprandial insulin responses generated a higher postprandial leucine responses compared to participants with lower insulin responses. CONCLUSIONS: The study demonstrated complex changes from catabolic to anabolic metabolism after a meal and indicated that the extent of postprandial responses is different between individuals with high and low insulin response.
Assuntos
Refeições , Metaboloma/fisiologia , Idoso , Aminoácidos/sangue , Glicemia/análise , Pão , Carnitina/análogos & derivados , Carnitina/sangue , Carboidratos da Dieta/administração & dosagem , Gorduras na Dieta/administração & dosagem , Fibras na Dieta/administração & dosagem , Proteínas Alimentares/administração & dosagem , Metabolismo Energético , Jejum , Feminino , Glicólise , Humanos , Insulina/sangue , Corpos Cetônicos/sangue , Leucina/sangue , Pessoa de Meia-Idade , Fosfolipídeos/sangue , Pós-Menopausa , Período Pós-Prandial , TriticumRESUMO
Changes in serum metabolic profile after the intake of different food products (e.g., bread) can provide insight into their interaction with human metabolism. Postprandial metabolic responses were compared after the intake of refined wheat (RWB), whole-meal rye (WRB), and refined rye (RRB) breads. In addition, associations between the metabolic profile in fasting serum and the postprandial concentration of insulin in response to different breads were investigated. Nineteen postmenopausal women with normal fasting glucose and normal glucose tolerance participated in a randomized, controlled, crossover meal study. The test breads, RWB (control), RRB, and WRB, providing 50 g of available carbohydrate, were each served as a single meal. The postprandial metabolic profile was measured using nuclear magnetic resonance and targeted LC-mass spectrometry and was compared between different breads using ANOVA and multivariate models. Eight amino acids had a significant treatment effect (P < 0.01) and a significant treatment × time effect (P < 0.05). RWB produced higher postprandial concentrations of leucine (geometric mean: 224; 95% CI: 196, 257) and isoleucine (mean ± SD: 111 ± 31.5) compared with RRB (geometric mean: 165; 95% CI: 147, 186; mean ± SD: 84.2 ± 22.9) and WRB (geometric mean: 190; 95% CI: 174, 207; mean ± SD: 95.8 ± 17.3) at 60 min respectively (P < 0.001). In addition, 2 metabolic subgroups were identified using multivariate models based on the association between fasting metabolic profile and the postprandial concentration of insulin. Women with higher fasting concentrations of leucine and isoleucine and lower fasting concentrations of sphingomyelins and phosphatidylcholines had higher insulin responses despite similar glucose concentration after all kinds of bread (cross-validated ANOVA, P = 0.048). High blood concentration of branched-chain amino acids, i.e., leucine and isoleucine, has been associated with the increased risk of diabetes, which suggests that additional consideration should be given to bread proteins in understanding the beneficial health effects of different kinds of breads. The present study suggests that the fasting metabolic profile can be used to characterize the postprandial insulin demand in individuals with normal glucose metabolism that can be used for establishing strategies for the stratification of individuals in personalized nutrition.
Assuntos
Pão , Jejum , Insulina/sangue , Metaboloma , Pós-Menopausa/fisiologia , Período Pós-Prandial/fisiologia , Idoso , Glicemia/metabolismo , Estudos Cross-Over , Feminino , Humanos , Isoleucina/sangue , Leucina/sangue , Pessoa de Meia-Idade , Análise Multivariada , Fosfatidilcolinas/sangue , Secale/química , Esfingomielinas/sangue , Triticum/químicaRESUMO
BACKGROUND: Epidemiological studies have consistently shown that whole grain (WG) cereals can protect against the development of chronic diseases, but the underlying mechanism is not fully understood. Among WG products, WG rye is considered even more potent because of its unique discrepancy in postprandial insulin and glucose responses known as the rye factor. In this study, an NMR-based metabolomics approach was applied to study the metabolic effects of WG rye as a tool to determine the beneficial effects of WG rye on human health. METHODS: Thirty-three postmenopausal Finnish women with elevated serum total cholesterol (5.0-8.5 mmol/L) and BMI of 20-33 kg/m² consumed a minimum of 20% of their daily energy intake as high fiber WG rye bread (RB) or refined wheat bread (WB) in a randomized, controlled, crossover design with two 8-wk intervention periods separated by an 8-wk washout period. At the end of each intervention period, fasting serum was collected for NMR-based metabolomics and the analysis of cholesterol fractions. Multilevel partial least squares discriminant analysis was used for paired comparisons of multivariate data. RESULTS: The metabolomics analysis of serum showed lower leucine and isoleucine and higher betaine and N,N-dimethylglycine levels after RB than WB intake. To further investigate the metabolic effects of RB, the serum cholesterol fractions were measured. Total- and LDL-cholesterol levels were higher after RB intake than after WB (p<0.05). CONCLUSIONS: This study revealed favorable shifts in branched amino acid and single carbon metabolism and an unfavorable shift in serum cholesterol levels after RB intake in postmenopausal women, which should be considered for evaluating health beneficial effects of rye products.
Assuntos
Pão , Fibras na Dieta/uso terapêutico , Hipercolesterolemia/dietoterapia , Secale/química , Sementes/química , Aminoácidos de Cadeia Ramificada/sangue , Betaína/sangue , Índice de Massa Corporal , Pão/efeitos adversos , Pão/análise , Estudos Cross-Over , Diabetes Mellitus Tipo 2/epidemiologia , Diabetes Mellitus Tipo 2/etiologia , Diabetes Mellitus Tipo 2/prevenção & controle , Fibras na Dieta/efeitos adversos , Fibras na Dieta/análise , Feminino , Finlândia/epidemiologia , Humanos , Hipercolesterolemia/complicações , Hipercolesterolemia/metabolismo , Hipercolesterolemia/fisiopatologia , Espectroscopia de Ressonância Magnética , Metabolômica/métodos , Pessoa de Meia-Idade , Sobrepeso/complicações , Sobrepeso/fisiopatologia , Pós-Menopausa , Período Pós-Prandial , Risco , Sarcosina/análogos & derivados , Sarcosina/sangue , Secale/efeitos adversos , Sementes/efeitos adversosRESUMO
[This corrects the article DOI: 10.3389/fmolb.2022.931946.].
RESUMO
Sustainability issues arise when using fish oil and vegetable oils in fish feed production for aquaculture purposes. Microbial production of single cell oil is a potential alternative as a lipid ingredient in the production of fish feed. In this study, we replaced the vegetable oils with the oleaginous yeast R. toruloides biomass in the diet of Arctic char (S. alpinus) and investigated the effects on health and composition. Measurement of fish growth parameters showed a higher liver weight and hepatosomatic index in the experimental group of fish fed partly with yeast biomass compared to a control group fed a diet with vegetable oils. No significant differences in the lipid content of muscle and liver tissues were found. The fatty acid profiles in the muscle of both fish groups were similar while the experimental fish group had a higher amount of monounsaturated fatty acids in the liver. Histology of livers showed no significant difference in the number of lipid droplets. The size of hepatic lipid droplets seemed to be related to liver fat content. Quantification of metabolites in the liver revealed no differences between the fish groups while plasma metabolites involved in energy pathways such as alanine, 3-hydroxybutyrate, creatinine, serine, betaine, and choline were significantly higher in the experimental fish group.
RESUMO
Metabolomics is an approach in which the profiles of metabolites in different tissues and/or biofluids are investigated to understand the changes induced following a modulation. We used this approach to investigate the biochemical effects of α-tocopherol in the liver using a rat model. Rats (21-day-old) were fed either an α-tocopherol-sufficient control (n = 10) or an α-tocopherol-deficient (n = 10) diet for 2 months before sacrifice. Livers were homogenized in methanol-chloroform-water (3 : 1 : 1, v/v/v), and the polar phase extracts of the liver samples were analyzed using (1) H NMR. Multivariate statistical analysis of the data was performed using principal component analysis and orthogonal partial least squares-discriminant analysis. Identification of (1) H NMR signals was performed primarily using the Human Metabolome Database, Biological Magnetic Resonance Data Bank and previous literature, and confirmed by spiking with metabolites and applying two-dimensional NMR. The statistical analysis revealed that α-tocopherol deficiency caused an increase in carnitine, choline, L-valine, L-lysine, tyrosine and inosine content and a reduction in glucose and uridine 5'-monophosphate content. Changes in carnitine and glucose suggest a possible shift in energy metabolism.
Assuntos
Fígado/metabolismo , Metaboloma , Metabolômica/métodos , alfa-Tocoferol/metabolismo , Animais , Humanos , Análise dos Mínimos Quadrados , Espectroscopia de Ressonância Magnética , Masculino , Ratos , Ratos Sprague-Dawley , Extratos de Tecidos/metabolismo , alfa-Tocoferol/sangueRESUMO
Prostate cancer (PC) is the most common cancer in the Western world and the second most important cancer causing male deaths, after lung cancer, in the United States and Britain. Lifestyle and dietary changes are recommended for men diagnosed with early-stage PC. It has been shown that a diet rich in whole grain (WG) rye reduces the progression of early-stage PC, but the underlying mechanism is not clear. This study sought to identify changes in the metabolic signature of plasma in patients with early-stage PC following intervention with a diet rich in WG rye and rye bran product (RP) compared with refined white wheat product (WP) as a tool for mechanistic investigation of the beneficial health effects of RP on PC progression. Seventeen PC patients received 485 g RP or WP in a randomized, controlled, crossover design during a period of 6 wk with a 2-wk washout period. At the end of each intervention period, plasma was collected after fasting and used for (1)H NMR-based metabolomics. Multilevel partial least squares discriminant analysis was used for paired comparisons of multivariate data. A metabolomics analysis of plasma showed an increase in 5 metabolites, including 3-hydroxybutyric acid, acetone, betaine, N,N-dimethylglycine, and dimethyl sulfone, after RP. To understand these metabolic changes, fasting plasma homocysteine, leptin, adiponectin, and glucagon were measured separately. The plasma homocysteine concentration was lower (P = 0.017) and that of leptin tended to be lower (P = 0.07) after RP intake compared to WP intake. The increase in plasma 3-hydroxybutyric acid and acetone after RP suggests a shift in energy metabolism from anabolic to catabolic status, which could explain some of the beneficial health effects of WG rye, i.e., reduction in prostate-specific antigen and reduced 24-h insulin secretion. In addition, the increase in betaine and N,N-dimethylglycine and the decrease in homocysteine show a favorable shift in homocysteine metabolism after RP intake.
Assuntos
Espectroscopia de Ressonância Magnética/métodos , Metaboloma/efeitos dos fármacos , Metabolômica/métodos , Preparações de Plantas/farmacologia , Neoplasias da Próstata/metabolismo , Secale/metabolismo , Ácido 3-Hidroxibutírico/sangue , Idoso , Idoso de 80 Anos ou mais , Betaína/sangue , Estudos Cross-Over , Dieta , Homocisteína/sangue , Humanos , Insulina/sangue , Masculino , Análise Multivariada , Antígeno Prostático Específico/sangue , Sarcosina/análogos & derivados , Sarcosina/sangue , Triticum/metabolismo , Estados UnidosRESUMO
OBJECTIVES: To assess the content and composition of fatty acids and tocopherols in commercially available oil capsules, and to assay thiobarbituric acid reactive substances (TBARS) as a marker of oxidation in these oils. METHODS: Fish and seal oil capsules were analyzed for their contents of long-chain omega-3 fatty acids (DPA, EPA, and DHA), tocopherols, and malondialdehyde. RESULTS: Large variations were found in the levels of EPA (6.5-40.9%), and DHA (8.1-26.4%), α-tocopherol (117-10282 µg/g), γ-tocopherol (406-2352 µg/g) and δ-tocopherol (127-978 µg/g). The level of malondialdehyde was very low in all capsules. CONCLUSION: The tested samples of oil capsules showed large variation in quality with respect to long-chain n-3 polyunsaturated fatty acids and tocopherols. The low levels of malondialdehyde indicated a successful oxidation protection strategy.
Assuntos
Suplementos Nutricionais/normas , Óleos de Peixe/normas , Tocoferóis/normas , Animais , Cápsulas , Cromatografia Gasosa , Cromatografia Líquida de Alta Pressão , Suplementos Nutricionais/análise , Ácidos Graxos Ômega-3/análise , Óleos de Peixe/administração & dosagem , Óleos de Peixe/análise , Peixes , Indicadores e Reagentes , Malondialdeído/análise , Oxirredução , Focas Verdadeiras , Espectrofotometria Ultravioleta , Suécia , Substâncias Reativas com Ácido Tiobarbitúrico/análise , Tocoferóis/administração & dosagem , Tocoferóis/análiseRESUMO
BACKGROUND: The protective role of mildly elevated bilirubin against CVD and diabetes mellitus type 2 (DMT2) is associated with a favorable lipid phenotype. As the mechanistic understanding of this protection in humans remains elusive, we aimed to assess the metabolomics profile of mildly hyperbilirubinemic (Gilbert's syndrome; GS) individuals especially targeting lipid catabolism. METHODS AND RESULTS: Using NMR serum metabolomics of 56 GS individuals and 56 age and gender-matched healthy controls, GS individuals demonstrated significantly greater concentrations of acetylcarnitine (+20%, pâ¯<â¯0.001) and the ketone bodies, 3-hydroxybutyric acid (+132%, pâ¯<â¯0.001), acetoacetic acid (+95%, pâ¯<â¯0.001) and acetone (+46%, pâ¯<â¯0.001). Metabolites associated with an increased mitochondrial lipid metabolism such as citrate (+15%, pâ¯<â¯0.001), anaplerotic amino acid intermediates and creatinine were significantly greater and creatine significantly reduced in GS individuals. Stimulators of lipid catabolism including AMPK (+59%, pâ¯<â¯0.001), pPPARα (+24%, pâ¯<â¯0.001) and T3 (+9%, pâ¯=â¯0.009) supported the metabolomics data while concomitantly blood glucose and insulin (-33%, pâ¯=â¯0.002) levels were significantly reduced. We further showed that the increased lipid catabolism partially mediates the favorable lipid phenotype (lower triglycerides) of GS individuals. Increased trimethylamine (+35%, pâ¯<â¯0.001) indicated changes in trimethylamine metabolism, an emerging predictor of metabolic health. CONCLUSION: We showed an enhanced lipid catabolism in mildly hyperbilirubinemic individuals, novel evidence as to why these individuals are leaner and protected against chronic metabolic diseases emphasizing bilirubin to be a promising future target in obese and dyslipidemia patients.
Assuntos
Bilirrubina/sangue , Doença de Gilbert/sangue , Metabolismo dos Lipídeos/fisiologia , Metaboloma/fisiologia , Adulto , Feminino , Humanos , Masculino , Metabolômica , Pessoa de Meia-Idade , Adulto JovemRESUMO
SCOPE: Ingestion of rye bread leads to lower postprandial plasma insulin concentrations than wheat bread ingestion, but most often not too different glucose profiles. The mechanism behind this discrepancy is still largely unknown. This study investigates whether glucose kinetics may explain the observed discrepancy. METHODS AND RESULTS: Nine healthy men participated in a crossover study, eating 50 g of available carbohydrates as either refined wheat (WB) or traditional wholemeal rye bread (WMR) during d-[6,6-2 H2 ]glucose infusion. Labeled glucose enrichment is measured by an HPLC-TOF-MS method. The calculated rate of glucose appearance (RaE) is significantly lower after ingestion of WMR during the initial 15 min postprandial period. Additionally, the 0-90 min RaE area under the curve (AUC) is significantly lower after ingestion of WMR, as is plasma gastric inhibitory polypeptide (GIP) at 60 and 90 min. Postprandial glycemic responses do not differ between the breads. Postprandial insulin is lower after ingestion of WMR at 45 and 60 min, as is the 0-90 min AUC. CONCLUSION: Ingestion of WMR elicits a lower rate of glucose appearance into the bloodstream compared with WB. This may explain the lower insulin response observed after rye bread ingestion, commonly known as the rye factor.
Assuntos
Glicemia/metabolismo , Pão , Insulina/sangue , Secale , Triticum , Adolescente , Adulto , Glicemia/análise , Peptídeo C/sangue , Cromatografia Líquida de Alta Pressão , Estudos Cross-Over , Polipeptídeo Inibidor Gástrico/sangue , Glucagon/sangue , Peptídeo 1 Semelhante ao Glucagon/sangue , Humanos , Masculino , Espectrometria de Massas , Período Pós-Prandial/fisiologiaRESUMO
Public health authorities recommend all fertile women to increase their folate intake to 400 µg/d by eating folate-rich foods or by taking a folic acid supplement to protect against neural tube defects. In a previous study it was shown that folate-rich foods improved folate blood status as effectively as folic acid supplementation. The aim of the present study was to investigate, using NMR metabolomics, the effects of an intervention with a synthetic folic acid supplement v. native food folate on the profile of plasma metabolites. Healthy women with normal folate status received, in parallel, 500 µg/d synthetic folic acid from a supplement (n 18), 250 µg/d folate from intervention foods (n 19), or no additional folate (0 µg/d) through a portion of apple juice (n 20). The metabolic profile of plasma was measured using 1H-NMR in fasted blood drawn at baseline and after 12 weeks of intervention. Metabolic differences between the groups at baseline and after intervention were assessed using a univariate statistical approach (P ≤ 0·001, Bonferroni-adjusted significance level). At baseline, the groups showed no significant differences in measured metabolite concentrations. After intervention, eight metabolites, of which six (glycine, choline, betaine, formate, histidine and threonine) are related to one-carbon metabolism, were identified as discriminative metabolites. The present study suggests that different folate forms (synthetic v. natural) may affect related one-carbon metabolites differently.
RESUMO
Mastitis is an inflammatory condition of the mammary tissue and represents a major problem for the dairy industry worldwide. The present study was undertaken to study how experimentally induced acute bovine mastitis affects inflammatory parameters and changes in the metabolome. To this end, we induced experimental mastitis in nine cows by intramammary infusion of 100 µg purified Escherichia coli lipopolysaccharide (LPS) followed by kinetic assessments of cytokine responses (by enzyme-linked immunosorbent assay), changes in the metabolome (assessed by nuclear magnetic resonance), clinical parameters (heat, local pain perception, redness, swelling, rectal temperature, clot formation, and color changes in the milk), and milk somatic cell counts, at several time points post LPS infusion. Intramammary LPS infusion induced clinical signs of mastitis, which started from 2 h post infusion and had returned to normal levels within 24-72 h. Milk changes were seen with a delay compared with the clinical signs and persisted for a longer time. In parallel, induction of IL-6 and TNF-α were seen in milk, and there was also a transient elevation of plasma IL-6 whereas plasma TNF-α was not significantly elevated. In addition, a robust increase in CCL2 was seen in the milk of LPS-infused cows, whereas G-CSF, CXCL1, and histamine in milk were unaffected. By using a metabolomics approach, a transient increase of plasma lactose was seen in LPS-induced cows. In plasma, significant reductions in ketone bodies (3-hydroxybutyrate and acetoacetate) and decreased levels of short-chain fatty acids, known to be major products released from the gut microbiota, were observed after LPS infusion; a profound reduction of plasma citrate was also seen. Intramammary LPS infusion also caused major changes in the milk metabolome, although with a delay in comparison with plasma, including a reduction of lactose. We conclude that the LPS-induced acute mastitis rapidly affects the plasma metabolome and cytokine induction with similar kinetics as the development of the clinical signs, whereas the corresponding effects in milk occurred with a delay.