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1.
J Proteome Res ; 17(10): 3492-3502, 2018 10 05.
Artigo em Inglês | MEDLINE | ID: mdl-30183320

RESUMO

The application of metabolic phenotyping to epidemiological studies involving thousands of biofluid samples presents a challenge for the selection of analytical platforms that meet the requirements of high-throughput precision analysis and cost-effectiveness. Here direct infusion-nanoelectrospray (DI-nESI) was compared with an ultra-performance liquid chromatography (UPLC)-high-resolution mass spectrometry (HRMS) method for metabolic profiling of an exemplary set of 132 human urine samples from a large epidemiological cohort. Both methods were developed and optimized to allow the simultaneous collection of high-resolution urinary metabolic profiles and quantitative data for a selected panel of 35 metabolites. The total run time for measuring the sample set in both polarities by UPLC-HRMS was 5 days compared with 9 h by DI-nESI-HRMS. To compare the classification ability of the two MS methods, we performed exploratory analysis of the full-scan HRMS profiles to detect sex-related differences in biochemical composition. Although metabolite identification is less specific in DI-nESI-HRMS, the significant features responsible for discrimination between sexes were mostly the same in both MS-based platforms. Using the quantitative data, we showed that 10 metabolites have strong correlation (Pearson's r > 0.9 and Passing-Bablok regression slope of 0.8-1.3) and good agreement assessed by Bland-Altman plots between UPLC-HRMS and DI-nESI-HRMS and thus can be measured using a cheaper and less sample- and time-consuming method. A further twenty metabolites showed acceptable correlation between the two methods with only five metabolites showing weak correlation (Pearson's  r < 0.4) and poor agreement due to the overestimation of the results by DI-nESI-HRMS.


Assuntos
Cromatografia Líquida de Alta Pressão/métodos , Metaboloma , Metabolômica/métodos , Espectrometria de Massas por Ionização por Electrospray/métodos , Espectrometria de Massas em Tandem/métodos , Adulto , Alanina/urina , Creatina/urina , Creatinina/urina , Feminino , Humanos , Hipertensão/metabolismo , Hipertensão/urina , Ácido Láctico/urina , Masculino , Pessoa de Meia-Idade , Nanotecnologia/métodos , Reprodutibilidade dos Testes
2.
J Proteome Res ; 17(9): 3184-3194, 2018 09 07.
Artigo em Inglês | MEDLINE | ID: mdl-30024170

RESUMO

Ureteral obstruction will lead clinically to hydronephrosis, which may further develop into partial or complete loss of kidney function and even cause permanent histological damage. However, there is little knowledge of metabolic responses during the obstructed process and its recoverability. In this study, a complete unilateral ureteral obstruction (CUUO) model was established in the rabbit, and 1H NMR-based metabolomic analysis of urine was used to reveal the metabolic perturbations in rabbits caused by CUUO and the metabolic recovery after the CUUO was relieved. Univariate and multivariate statistical analyses were used to identify metabolic characteristics. The gradually decreased levels of 3-hydroxykynurenine, 3-methylhistidine, creatinine, guanidoacetate, meta- and para-hydroxyphenylacetate, and phenylacetylglycine and the gradually increased levels of acetate, alanine, citrate, glycine, lactate, and methionine in urine could be regarded as potential biomarkers for the occurrence and severity of ureteral obstruction. And the reduced levels of 3-methylhistidine, creatinine, guanidoacetate, hippurate, meta-hydroxyphenylacetate, and methylguanidine and the elevated levels of 2-aminoisobutyrate, acetylcholine, citrate, lactate, lysine, valine, and α-ketoglutarate in urine compared with the obstructed level could characterize the metabolic recovery of ureteral obstruction. Our results depicted the disturbed biochemical pathways involved in ureteral obstruction and demonstrated the practicability of recovering renal functions for the patients with severe hydronephrosis in clinical practice by removing causes for obstruction.


Assuntos
Hidronefrose/urina , Cinurenina/análogos & derivados , Metaboloma , Metilistidinas/urina , Obstrução Ureteral/urina , Ácido Acético/urina , Alanina/urina , Análise de Variância , Animais , Biomarcadores/urina , Ácido Cítrico/urina , Creatinina/urina , Modelos Animais de Doenças , Glicina/análogos & derivados , Glicina/urina , Hidronefrose/diagnóstico , Hidronefrose/patologia , Cinurenina/urina , Ácido Láctico/urina , Espectroscopia de Ressonância Magnética , Masculino , Metionina/urina , Fenilacetatos/urina , Coelhos , Ureter/metabolismo , Ureter/patologia , Ureter/cirurgia , Obstrução Ureteral/diagnóstico , Obstrução Ureteral/patologia
3.
J Proteome Res ; 17(3): 1278-1289, 2018 03 02.
Artigo em Inglês | MEDLINE | ID: mdl-29424227

RESUMO

This nuclear magnetic resonance metabolomics study compared the influence of two different central Portugal exposomes, one of which comprised an important source of pollutants (the Estarreja Chemical Complex, ECC), on the urinary metabolic trajectory of a cohort of healthy pregnant women (total n = 107). An exposome-independent description of pregnancy metabolism was found to comprise a set of 18 metabolites reflecting expected changes in branched-chain amino acid catabolism and hormone and lipid metabolisms. In addition, a set of small changes in some metabolites was suggested to be exposome-dependent and characteristic of pregnant subjects from the Estarreja region. These results suggested that the Estarreja exposome may impact to a very low extent pregnancy metabolism, inducing slight changes in amino acid metabolism (alanine, glycine, and 3-hydroxyisobutyrate, possibly involved in valine metabolism), tricarboxylic acid (TCA) cycle (cis-aconitate), diet, or gut microflora (furoylglycine) as well as allantoin, 2-hydroxyisobutyrate, and an unassigned resonance at δ 8.45. Furthermore, the urine of Estarreja subjects was found to generally contain higher levels of 4-hydroxyphenylacetate and lower levels of citrate. However, out of the above metabolites, only glycine and citrate seemed to correlate with the proximity to the ECC, with slightly relative higher levels of these compounds found for subjects living closer to the ECC. This suggested possible small effects of local pollutants on energy metabolism, with the remaining exposome-dependent metabolite changes most probably originating from other aspects of the local exposome such as diet and lifestyle. Despite the limitation of this study regarding the unavailability of objective environmental parameters for the period under study, our results confirm the usefulness of metabolomics of human urine to gauge exposome effects on human health and, particularly, during pregnancy.


Assuntos
Poluentes Atmosféricos/efeitos adversos , Metabolismo Energético/efeitos dos fármacos , Exposição Ambiental/efeitos adversos , Metaboloma , Ácido Aconítico/urina , Adulto , Alanina/urina , Alantoína/urina , Indústria Química , Ácido Cítrico/urina , Estudos de Coortes , Dieta/métodos , Feminino , Glicina/análogos & derivados , Glicina/urina , Humanos , Hidroxibutiratos/urina , Estilo de Vida , Espectroscopia de Ressonância Magnética , Fenilacetatos/urina , Gravidez , Espanha
4.
Auris Nasus Larynx ; 44(4): 381-389, 2017 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-27817938

RESUMO

OBJECTIVE: Idiopathic sudden sensorineural hearing loss is a frequent emergency, with unknown aetiology and usually treated with empiric therapy. Steroids represent the only validated treatment but prognosis is unpredictable and the possibility to select the patients who will not respond to steroids could avoid unnecessary treatments. Metabolomic profiling of the biofluids target the analysis of the final product of genic expression and enzymatic activity, defining the biochemical phenotype of a whole biologic system. METHODS: We studied the metabolomics of the urine of a cohort of patients with idiopathic sudden sensorineural hearing loss, correlating the metabolic profiles with the clinical outcomes. Metabolomic profiling of urine samples was performed by 1H Nuclear Magnetic Resonance spectroscopy in combination with multivariate statistical approaches. RESULTS: 26 patients were included in the study: 5 healthy controls, 13 patients who did not recover after treatment at 6 months while the remaining 8 patients recovered from the hearing loss. The orthogonal partial least square-discriminant analysis score plot showed a significant separation between the two groups, responders and non-responders after steroid therapy, R2Y of 0.83, Q2 of 0.38 and p value <0.05. The resulting metabolic profiles were characterized by higher levels of urinary B-Alanine, 3-hydroxybutyrate and Trimethylamine N-oxide, and lower levels of Citrate and Creatinine in patients with worst outcome. CONCLUSION: Idiopathic sudden sensorineural hearing loss is a specific disease with unclear systemic changes, but our data suggest that there are different types of this disorder or patients predisposed to effective action of steroids allowing the recover after treatment.


Assuntos
Ácido 3-Hidroxibutírico/urina , Alanina/urina , Perda Auditiva Súbita/urina , Metabolômica , Metilaminas/urina , Adolescente , Adulto , Idoso , Anticoagulantes/uso terapêutico , Audiometria de Tons Puros , Estudos de Casos e Controles , Criança , Ácido Cítrico/urina , Estudos de Coortes , Creatinina/urina , Enoxaparina/uso terapêutico , Feminino , Perda Auditiva Súbita/terapia , Humanos , Oxigenação Hiperbárica/métodos , Imunossupressores/uso terapêutico , Masculino , Pessoa de Meia-Idade , Pregnenodionas/uso terapêutico , Prognóstico , Estudos Prospectivos , Espectroscopia de Prótons por Ressonância Magnética , Urina/química , Adulto Jovem
5.
Nutrients ; 8(2): 96, 2016 Feb 17.
Artigo em Inglês | MEDLINE | ID: mdl-26901221

RESUMO

Wheat bran, and especially wheat aleurone fraction, are concentrated sources of a wide range of components which may contribute to the health benefits associated with higher consumption of whole-grain foods. This study used NMR metabolomics to evaluate urine samples from baseline at one and two hours postprandially, following the consumption of minimally processed bran, aleurone or control by 14 participants (7 Females; 7 Males) in a randomized crossover trial. The methodology discriminated between the urinary responses of control, and bran and aleurone, but not between the two fractions. Compared to control, consumption of aleurone or bran led to significantly and substantially higher urinary concentrations of lactate, alanine, N-acetylaspartate acid and N-acetylaspartylglutamate and significantly and substantially lower urinary betaine concentrations at one and two hours postprandially. There were sex related differences in urinary metabolite profiles with generally higher hippurate and citrate and lower betaine in females compared to males. Overall, this postprandial study suggests that acute consumption of bran or aleurone is associated with a number of physiological effects that may impact on energy metabolism and which are consistent with longer term human and animal metabolomic studies that used whole-grain wheat diets or wheat fractions.


Assuntos
Dieta , Fibras na Dieta/metabolismo , Período Pós-Prandial , Sementes/química , Triticum/química , Grãos Integrais/metabolismo , Adulto , Alanina/urina , Ácido Aspártico/análogos & derivados , Ácido Aspártico/urina , Betaína/urina , Ácido Cítrico/urina , Dipeptídeos/urina , Feminino , Manipulação de Alimentos , Hipuratos/urina , Humanos , Ácido Láctico/urina , Espectroscopia de Ressonância Magnética , Masculino , Metabolômica/métodos , Fatores Sexuais , Adulto Jovem
6.
J Ethnopharmacol ; 182: 80-9, 2016 Apr 22.
Artigo em Inglês | MEDLINE | ID: mdl-26899442

RESUMO

ETHNOPHARMACOLOGICAL RELEVANCE: Eurycoma longifolia (Tongkat Ali, TA) roots have been ethnically used as a remedy to boost male sexual desire, libido, energy and fertility. AIM OF THE STUDY: The study evaluated the effect of TA extracts with different quassinoid levels on rats sperm count and examined corresponding post-treatment urinary metabolic changes. MATERIALS AND METHODS: Twenty-four male Sprague-Dawley rats, categorized into 4 groups of 6 rats each, were orally administered for 48 days with water for the control (group 1), 125mg/kg of TA water extract (TAW, group 2), 125mg/kg of TA quassinoid-poor extract (TAQP, group 3) and 21mg/kg of TA quassinoid-rich extract (TAQR, group 4). Upon completion of the 48-day treatment, the urine samples were analyzed by NMR and the animals were subsequently sacrificed for sperm count analysis. The urine profiles were categorized according to sperm count level. RESULTS: The results showed that the sperm count in TAW- and TAQR-treated groups was significantly higher compared to the TAQP-administered and control groups. The orthogonal partial least squares discriminant analysis (OPLS-DA) model indicated a clear separation among the urine profiles with respect to sperm count level. Urine (1)H-NMR profiles of the high-sperm count group contained higher concentrations of trigonelline, alanine, benzoic acid and higher intensity of a signal at 3.42ppm, while ethanol was at higher concentration in the normal-sperm count group. CONCLUSIONS: The results proved the efficacy of quassinoids on sperm count increase in rats and provided quantitative markers in urine suitable for analysis of sperm profile and male fertility status.


Assuntos
Eurycoma , Metabolômica , Extratos Vegetais/farmacologia , Contagem de Espermatozoides , Administração Oral , Alanina/urina , Alcaloides/urina , Animais , Ácido Benzoico/urina , Biomarcadores/urina , Etanol/urina , Masculino , Espectroscopia de Prótons por Ressonância Magnética , Ratos , Ratos Sprague-Dawley
7.
Physiother Theory Pract ; 31(5): 354-61, 2015 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-25585514

RESUMO

Recently, low-level laser (light) therapy (LLLT) has been used to improve muscle performance. This study aimed to evaluate the effectiveness of near-infrared light-emitting diode therapy (LEDT) and its mechanisms of action to improve muscle performance in an elite athlete. The kinetics of oxygen uptake (VO2), blood and urine markers of muscle damage (creatine kinase--CK and alanine), and fatigue (lactate) were analyzed. Additionally, some metabolic parameters were assessed in urine using proton nuclear magnetic resonance spectroscopy ((1)H NMR). A LED cluster with 50 LEDs (λ = 850 nm; 50 mW 15 s; 37.5 J) was applied on legs, arms and trunk muscles of a single runner athlete 5 min before a high-intense constant workload running exercise on treadmill. The athlete received either Placebo-1-LEDT; Placebo-2-LEDT; or Effective-LEDT in a randomized double-blind placebo-controlled trial with washout period of 7 d between each test. LEDT improved the speed of the muscular VO2 adaptation (∼-9 s), decreased O2 deficit (∼-10 L), increased the VO2 from the slow component phase (∼+348 ml min(-1)), and increased the time limit of exercise (∼+589 s). LEDT decreased blood and urine markers of muscle damage and fatigue (CK, alanine and lactate levels). The results suggest that a muscular pre-conditioning regimen using LEDT before intense exercises could modulate metabolic and renal function to achieve better performance.


Assuntos
Luz , Terapia com Luz de Baixa Intensidade , Contração Muscular/efeitos da radiação , Músculo Esquelético/efeitos da radiação , Resistência Física/efeitos da radiação , Corrida , Adulto , Alanina/urina , Biomarcadores/sangue , Biomarcadores/urina , Creatina Quinase/sangue , Método Duplo-Cego , Humanos , Cinética , Ácido Láctico/sangue , Masculino , Fadiga Muscular/efeitos da radiação , Músculo Esquelético/metabolismo , Consumo de Oxigênio/efeitos da radiação , Espectroscopia de Prótons por Ressonância Magnética , Fatores de Tempo
8.
Pestic Biochem Physiol ; 116: 32-9, 2014 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-25454518

RESUMO

Metalaxyl [N-(2,6-dimethylphenyl)-N-(methoxyacetyl)-D,L-alaninemethylester] is a systemic fungicide widely used in agriculture. In this study, the enantioselective distribution, degradation and excretion of metalaxyl were investigated after oral gavage administration of rac-metalaxyl to mice. Concentration of metalaxyl and its enantiomers was determined by HPLC-MS/MS. The results showed that R-metalaxyl was much higher than S-metalaxyl in heart, liver, lung, urine and feces. As for the strong first pass effect, concentrations of metalaxyl in liver were much higher than those in other tissues. The total body clearance (CL) of metalaxyl in mice was 1.77 L h(-1 )kg(-1) and degradation half-lives of (t1/2) of S-metalaxyl and R-metalaxyl in liver were 2.2 h and 3.0 h, respectively. Such results indicated the enantioselectivity of metalaxyl lies in distribution, degradation and excretion processes in mice. Main metabolites were also determined and biotransformation reactions were hydroxylation, demethylation and didemethylation. Furthermore, metabolite concentrations in urine and feces were much higher than those in tissues. These results may have potential implications to predict toxicity and provide additional information associated with adverse health effects for risk assessment of metalaxyl.


Assuntos
Alanina/análogos & derivados , Fungicidas Industriais/química , Fungicidas Industriais/farmacocinética , Administração Oral , Alanina/sangue , Alanina/química , Alanina/farmacocinética , Alanina/urina , Animais , Cromatografia Líquida , Fezes/química , Fungicidas Industriais/sangue , Fungicidas Industriais/urina , Fígado/metabolismo , Pulmão/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos ICR , Miocárdio/metabolismo , Estereoisomerismo , Espectrometria de Massas em Tandem
9.
Ann Surg Oncol ; 21 Suppl 4: S736-42, 2014 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-25092158

RESUMO

BACKGROUND: Mass screening for gastric cancer (GC), particularly using endoscopy, may not be the most practical approach as a result of its high cost, lack of acceptance, and poor availability. Thus, novel markers that can be used in cost-effective diagnosis and noninvasive screening for GC are needed. METHODS: A total of 154 urine samples from GC patients and healthy individuals and 30 pairs of matched tumor and normal stomach tissues were collected. Multivariate analysis was performed on urinary and tissue metabolic profiles acquired using (1)H nuclear magnetic resonance and (1)H high-resolution magic angle spinning spectroscopy, respectively. In addition, metabolic profiling of urine from GC patients after curative surgery was performed. RESULTS: Multivariate statistical analysis showed significant separation in the urinary and tissue data of GC patients and healthy individuals. The metabolites altered in the urine of GC patients were related to amino acid and lipid metabolism, consistent with changes in GC tissue. In the external validation, the presence of GC (early or advanced) from the urine model was predicted with high accuracy, which showed much higher sensitivity than carbohydrate antigen 19-9 and carcinoembryonic antigen. Furthermore, 4-hydroxyphenylacetate, alanine, phenylacetylglycine, mannitol, glycolate, and arginine levels were significantly correlated with cancer T stage and, together with hypoxanthine level, showed a recovery tendency toward healthy controls in the postoperative samples compared to the preoperative samples. CONCLUSIONS: An urinary metabolomics approach may be useful for the effective diagnosis of GC.


Assuntos
Biomarcadores Tumorais/urina , Espectroscopia de Ressonância Magnética , Metaboloma , Neoplasias Gástricas/patologia , Neoplasias Gástricas/urina , Alanina/urina , Área Sob a Curva , Arginina/urina , Biomarcadores Tumorais/sangue , Antígeno CA-19-9/sangue , Antígeno Carcinoembrionário/sangue , Estudos de Casos e Controles , Glicina/análogos & derivados , Glicina/urina , Glicolatos/urina , Humanos , Hipoxantina/urina , Manitol/urina , Metabolômica , Estadiamento de Neoplasias , Fenilacetatos/urina , Curva ROC , Neoplasias Gástricas/diagnóstico , Neoplasias Gástricas/cirurgia , Resultado do Tratamento , Urinálise/métodos
10.
Dis Markers ; 35(5): 345-51, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-24191128

RESUMO

BACKGROUND: Metabolomics studies can quantitatively detect the dynamic metabolic response of living systems. OBJECTIVE: To detect urinary metabolomics after hepatic ischemia/reperfusion (I/R) injury induced by the Pringle maneuver using gas chromatography-mass spectrometry (GC-MS). METHODS: Male Sprague-Dawley rats (N = 80) were randomly divided into 4 groups (n = 20/group): sham operation, day 1, day 3, and day 5. Rats in the day 1, day 3, and day 5 groups underwent the Pringle maneuver. Serum alanine transaminase (ALT) and total bilirubin (TBIL) were measured, and hematoxylin and eosin (HE) staining of the liver tissue was performed. GC-MS was used to detect urinary metabolomics. RESULTS: Compared with the sham group, the serum ALT and TBIL levels at day 1 were significantly elevated (P < 0.01) and then decreased and reached close to normal levels at day 5. GC-MS detected 7 metabolites which had similar changes as those of liver tissue revealed by histological examination. Significant differences in lactic acid, pyruvic acid, alanine, serine, and glycerol-3-phosphate were found among the groups (P < 0.001). Principle component analysis showed that 7 metabolites distinguished the day 1 and day 3 groups from the sham group. CONCLUSIONS: Noninvasive urinary metabolomic analysis is a potential means for the early detection and diagnosis of hepatic I/R injury.


Assuntos
Fígado/patologia , Metaboloma , Traumatismo por Reperfusão/urina , Alanina/metabolismo , Alanina/urina , Alanina Transaminase/sangue , Animais , Bilirrubina/sangue , Cromatografia Gasosa-Espectrometria de Massas , Glicerofosfatos/metabolismo , Glicerofosfatos/urina , Ácido Láctico/metabolismo , Ácido Láctico/urina , Fígado/metabolismo , Masculino , Metabolômica , Ácido Pirúvico/metabolismo , Ácido Pirúvico/urina , Ratos , Ratos Sprague-Dawley , Traumatismo por Reperfusão/sangue , Traumatismo por Reperfusão/metabolismo , Serina/metabolismo , Serina/urina
11.
Pharmacology ; 92(3-4): 207-16, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-24136086

RESUMO

BACKGROUND/AIMS: Absorption, biotransformation and elimination of safinamide, an enantiomeric α-aminoamide derivative developed as an add-on therapy for Parkinson's disease patients, were studied in healthy volunteers administered a single oral dose of 400 mg (14)C safinamide methanesulphonate, labelled in metabolically stable positions. METHODS: Pharmacokinetics of the parent compound were investigated up to 96 h, of (14)C radioactivity up to 192/200 h post-dose. RESULTS/CONCLUSIONS: Maximum concentration was achieved at 1 h (plasma, median Tmax) for parent drug and at 7 and 1.5 h for plasma and whole blood (14)C radioactivity, respectively. Terminal half-lives were about 22 h for unchanged safinamide and 80 h for radioactivity. Safinamide deaminated acid and the N-dealkylated acid were identified as major metabolites in urine and plasma. In urine, the ß-glucuronide of the N-dealkylated acid and the monohydroxy safinamide were also characterized. In addition, the glycine conjugate of the N-dealkylated acid and 2-[4-hydroxybenzylamino]propanamide were tentatively identified as minor urinary metabolites.


Assuntos
Alanina/análogos & derivados , Benzilaminas/farmacocinética , Adulto , Alanina/sangue , Alanina/farmacocinética , Alanina/urina , Benzilaminas/sangue , Benzilaminas/urina , Fezes/química , Voluntários Saudáveis , Humanos , Masculino , Doença de Parkinson/tratamento farmacológico , Adulto Jovem
12.
Am J Clin Nutr ; 98(1): 136-45, 2013 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-23656904

RESUMO

BACKGROUND: Available data have indicated independent direct relations of dietary animal protein and meat to the blood pressure (BP) of individuals. OBJECTIVE: In this study, we aimed to assess whether BP is associated with the intake of dietary amino acids higher relatively in animal than in vegetable protein (alanine, arginine, aspartic acid, glycine, histidine, lysine, methionine, and threonine). DESIGN: The study was a cross-sectional epidemiologic study that involved 4680 persons aged 40-59 y from 17 random population samples in the People's Republic of China, Japan, the United Kingdom, and the United States. BP was measured 8 times at 4 visits; dietary data (83 nutrients and 18 amino acids) were from four 24-h dietary recalls and two 24-h urine collections. RESULTS: Dietary glycine and alanine (the percentage of total protein intake) were considered singly related directly to BP; with these 2 amino acids together in regression models (from model 1, which was controlled for age, sex, and sample, to model 5, which was controlled for 16 possible confounders), glycine, but not alanine, was significantly related to BP. Estimated average BP differences associated with a 2-SD higher glycine intake (0.71 g/24 h) were 2.0-3.0-mm Hg systolic BP (z = 2.97-4.32) stronger in Western than in East Asian participants. In Westerners, meat was the main dietary source of glycine but not in East Asians (Chinese: grains/flour and rice/noodles; Japanese: fish/shellfish and rice/noodles). CONCLUSION: Dietary glycine may have an independent adverse effect on BP, which possibly contributes to direct relations of animal protein and meat to BP.


Assuntos
Pressão Sanguínea/fisiologia , Proteínas na Dieta/administração & dosagem , Glicina/administração & dosagem , Micronutrientes/administração & dosagem , Adulto , Alanina/administração & dosagem , Alanina/efeitos adversos , Alanina/urina , Determinação da Pressão Arterial , China/epidemiologia , Estudos Transversais , Dieta , Proteínas na Dieta/urina , Feminino , Glicina/efeitos adversos , Glicina/urina , Humanos , Hipertensão/epidemiologia , Hipertensão/etiologia , Japão/epidemiologia , Modelos Lineares , Masculino , Micronutrientes/efeitos adversos , Pessoa de Meia-Idade , Inquéritos e Questionários , Reino Unido/epidemiologia , Estados Unidos/epidemiologia
13.
Anal Bioanal Chem ; 405(10): 3153-8, 2013 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-23354581

RESUMO

Since sarcosine and D,L-alanine co-elute on reversed-phase high-performance liquid chromatography (HPLC) columns and the tandem mass spectrometer cannot differentiate them due to equivalent parent and fragment ions, derivatization is often required for analysis of sarcosine in LC/MS systems. This study offers an alternative to derivatization by employing partial elimination of sarcosine by enzymatic oxidation. The decrease in apparent concentration from the traditionally merged sarcosine-alanine peak associated with the enzymatic elimination has been shown to be proportional to the total sarcosine present (R(2) = 0.9999), allowing for determinations of urinary sarcosine. Sarcosine oxidase was shown to eliminate only sarcosine in the presence of D,L-alanine, and was consequently used as the selective enzyme. This newly developed technique has a method detection limit of 1 µg/L (parts per billion) with a linear range of 3 ppb-1 mg/L (parts per million) in urine matrices. The method was further validated through spiked recoveries of real urine samples, as well as the analysis of 35 real urine samples. The average recoveries for low, middle, and high sarcosine concentration spikes were 111.7, 90.8, and 90.1 %, respectively. In conclusion, this simple enzymatic approach coupled with HPLC/MS/MS is able to resolve sarcosine from D,L-alanine leading to underivatized quantification of sarcosine.


Assuntos
Alanina/análise , Cromatografia Líquida de Alta Pressão/métodos , Sarcosina Oxidase/química , Sarcosina/química , Espectrometria de Massas em Tandem/métodos , Alanina/urina , Cromatografia de Fase Reversa/métodos , Humanos , Masculino , Oxirredução , Neoplasias da Próstata/urina , Sarcosina/urina
14.
Mol Cell Proteomics ; 12(1): 207-14, 2013 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-23111923

RESUMO

Major depressive disorder (MDD) is a widespread and debilitating mental disorder. However, there are no biomarkers available to aid in the diagnosis of this disorder. In this study, a nuclear magnetic resonance spectroscopy-based metabonomic approach was employed to profile urine samples from 82 first-episode drug-naïve depressed subjects and 82 healthy controls (the training set) in order to identify urinary metabolite biomarkers for MDD. Then, 44 unselected depressed subjects and 52 healthy controls (the test set) were used to independently validate the diagnostic generalizability of these biomarkers. A panel of five urinary metabolite biomarkers-malonate, formate, N-methylnicotinamide, m-hydroxyphenylacetate, and alanine-was identified. This panel was capable of distinguishing depressed subjects from healthy controls with an area under the receiver operating characteristic curve (AUC) of 0.81 in the training set. Moreover, this panel could classify blinded samples from the test set with an AUC of 0.89. These findings demonstrate that this urinary metabolite biomarker panel can aid in the future development of a urine-based diagnostic test for MDD.


Assuntos
Transtorno Depressivo Maior/diagnóstico , Transtorno Depressivo Maior/urina , Metaboloma , Metabolômica/métodos , Adulto , Alanina/urina , Biomarcadores/urina , Feminino , Formiatos/urina , Ácido Homovanílico/análogos & derivados , Ácido Homovanílico/urina , Humanos , Masculino , Malonatos/urina , Niacinamida/análogos & derivados , Niacinamida/urina , Ressonância Magnética Nuclear Biomolecular , Fenilacetatos
15.
Yao Xue Xue Bao ; 48(11): 1733-7, 2013 Nov.
Artigo em Chinês | MEDLINE | ID: mdl-24475714

RESUMO

To investigate the intervention effects of Morinda officinalis How. on 'Kidney-yang deficiency syndrome' induced by hydrocortisone in rats, the metabolic profiles of rat urine were characterized using proton nuclear magnetic resonance and principal component analysis (PCA) was applied to study the trajectory of urinary metabolic phenotype of rats with 'Kidney-yang deficiency syndrome' under administration of M. officinalis at different time points. Meanwhile, the intervention effects of M. officinalis on urinary metabolic potential biomarkers associated with 'Kidney-yang deficiency syndrome' were also discussed. The experimental results showed that in accordance to the increased time of administration, an obvious tendency was observed that clustering of the treatment group moved gradually closed to that of the control group. Eight potential biomarkers including citrate, succinate, alpha-ketoglutarate, lactate, betaine, sarcosine, alanine and taurine were definitely up- or down-regulated. In conclusion, the effectiveness of M. oficinalis on 'Kidney-yang deficiency syndrome' is proved using the established metabonomic method and the regulated metabolic pathways involve energy metabolism, transmethylation and transportation of amine. Meanwhile, the administration of M. officinalis can alleviate the kidney impairment induced by 'Kidney-yang deficiency syndrome'.


Assuntos
Biomarcadores/urina , Medicamentos de Ervas Chinesas/farmacologia , Nefropatias/urina , Morinda/química , Deficiência da Energia Yang/urina , Alanina/urina , Animais , Betaína/urina , Ácido Cítrico/urina , Medicamentos de Ervas Chinesas/isolamento & purificação , Hidrocortisona , Ácidos Cetoglutáricos/urina , Nefropatias/induzido quimicamente , Ácido Láctico/urina , Espectroscopia de Ressonância Magnética , Masculino , Metabolômica/métodos , Plantas Medicinais/química , Análise de Componente Principal , Distribuição Aleatória , Ratos , Ratos Sprague-Dawley , Sarcosina/urina , Ácido Succínico/urina , Taurina/urina , Deficiência da Energia Yang/induzido quimicamente
16.
Mol Cell Proteomics ; 11(8): 370-80, 2012 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-22505723

RESUMO

Metabolomics is a powerful new technology that allows for the assessment of global metabolic profiles in easily accessible biofluids and biomarker discovery in order to distinguish between diseased and nondiseased status information. Deciphering the molecular networks that distinguish diseases may lead to the identification of critical biomarkers for disease aggressiveness. However, current diagnostic methods cannot predict typical Jaundice syndrome (JS) in patients with liver disease and little is known about the global metabolomic alterations that characterize JS progression. Emerging metabolomics provides a powerful platform for discovering novel biomarkers and biochemical pathways to improve diagnostic, prognostication, and therapy. Therefore, the aim of this study is to find the potential biomarkers from JS disease by using a nontarget metabolomics method, and test their usefulness in human JS diagnosis. Multivariate data analysis methods were utilized to identify the potential biomarkers. Interestingly, 44 marker metabolites contributing to the complete separation of JS from matched healthy controls were identified. Metabolic pathways (Impact-value≥0.10) including alanine, aspartate, and glutamate metabolism and synthesis and degradation of ketone bodies were found to be disturbed in JS patients. This study demonstrates the possibilities of metabolomics as a diagnostic tool in diseases and provides new insight into pathophysiologic mechanisms.


Assuntos
Biomarcadores/urina , Icterícia/urina , Hepatopatias/urina , Metabolômica/métodos , Adulto , Alanina/metabolismo , Alanina/urina , Ácido Aspártico/metabolismo , Ácido Aspártico/urina , Biomarcadores/metabolismo , Cromatografia Líquida de Alta Pressão , Progressão da Doença , Feminino , Ácido Glutâmico/metabolismo , Ácido Glutâmico/urina , Humanos , Icterícia/diagnóstico , Icterícia/metabolismo , Corpos Cetônicos/metabolismo , Corpos Cetônicos/urina , Hepatopatias/metabolismo , Masculino , Espectrometria de Massas , Redes e Vias Metabólicas , Pessoa de Meia-Idade , Análise Multivariada , Prognóstico , Reprodutibilidade dos Testes , Sensibilidade e Especificidade
17.
Drug Metab Dispos ; 39(5): 891-903, 2011 May.
Artigo em Inglês | MEDLINE | ID: mdl-21289073

RESUMO

Brivanib [(R)-1-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-5-methylpyrrolo[1,2,4]triazin-6-yloxy)propan-2-ol, BMS-540215] is a potent and selective dual inhibitor of vascular endothelial growth factor (VEGF) and fibroblast growth factor (FGF) signaling pathways. Its alanine prodrug, brivanib alaninate [(1R,2S)-2-aminopropionic acid 2-[4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-5-methylpyrrolo[2,1-f][1,2,4]triazin-6-yloxy]-1-methylethyl ester, BMS-582664], is currently under development as an oral agent for the treatment of cancer. This study describes the in vivo biotransformation of brivanib after a single oral dose of [(14)C]brivanib alaninate to intact rats, bile duct-cannulated (BDC) rats, intact monkeys, BDC monkeys, and humans. Fecal excretion was the primary route of elimination of drug-derived radioactivity in animals and humans. In BDC rats and monkeys, the majority of radioactivity was excreted in bile. Brivanib alaninate was rapidly and completely converted via hydrolysis to brivanib in vivo. The area under the curve from zero to infinity of brivanib accounted for 14.2 to 54.3% of circulating radioactivity in plasma in animals and humans, suggesting that metabolites contributed significantly to the total drug-related radioactivity. In plasma from animals and humans, brivanib was a prominent circulating component. All the metabolites that humans were exposed to were also present in toxicological species. On the basis of metabolite exposure and activity against VEGF and FGF receptors of the prominent human circulating metabolites, only brivanib is expected to contribute to the pharmacological effects in humans. Unchanged brivanib was not detected in urine or bile samples, suggesting that metabolic clearance was the primary route of elimination. The primary metabolic pathways were oxidative and conjugative metabolism of brivanib.


Assuntos
Alanina/análogos & derivados , Antineoplásicos/metabolismo , Triazinas/metabolismo , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/antagonistas & inibidores , Administração Oral , Alanina/administração & dosagem , Alanina/metabolismo , Alanina/farmacocinética , Alanina/urina , Animais , Antineoplásicos/administração & dosagem , Antineoplásicos/farmacocinética , Antineoplásicos/urina , Bile/metabolismo , Biotransformação , Fezes , Humanos , Macaca fascicularis , Masculino , Neoplasias/tratamento farmacológico , Ensaio Radioligante , Ratos , Ratos Sprague-Dawley , Triazinas/administração & dosagem , Triazinas/farmacocinética , Triazinas/urina
18.
J Chromatogr B Analyt Technol Biomed Life Sci ; 879(29): 3184-9, 2011 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-20851062

RESUMO

A fully automated two-dimensional HPLC system combining a microbore-ODS column and a narrowbore-enantioselective column was designed and validated, and the amounts of D-serine (D-Ser) and D-alanine (D-Ala) in various tissues and physiological fluids of Long-Evans agouti/SENDAI (LEA/Sen) rats lacking D-amino-acid oxidase (DAO) were determined. Intra- and inter-day precision was less than 4.3% and accuracy ranged between 99.9 and 104%. LEA/Sen rats were reported to lack DAO in their kidneys and expected to be a novel mutant animal lacking DAO, however, the amounts of D-amino acids in the LEA/Sen rats have not been investigated. In the present study, the intrinsic amounts of D-Ser and D-Ala, which are neuromodulators of the N-methyl-D-aspartate (NMDA) receptors, were determined in seven brain tissues, four peripheral tissues, plasma and urine of the LEA/Sen rats and compared to those of the control (Wistar and SD) rats having normal DAO activity. The levels of D-Ser in the tissues and physiological fluids of the LEA/Sen rats were significantly higher than those of the Wistar and SD rats except for the frontal brain regions. Concerning D-Ala, the amounts in the tissues and physiological fluids of the LEA/Sen rats were drastically increased compared to those of the Wistar and SD rats. These results indicate that the intrinsic amounts of D-Ser and D-Ala in the tissues of rats are regulated by DAO, and that LEA/Sen rats would be useful for the study of NMDA receptor-related diseases in which DAO is implicated.


Assuntos
Alanina/análise , Química Encefálica , Cromatografia Líquida de Alta Pressão/métodos , D-Aminoácido Oxidase/metabolismo , Serina/análise , Alanina/sangue , Alanina/química , Alanina/urina , Animais , D-Aminoácido Oxidase/genética , Modelos Animais de Doenças , Rim/química , Masculino , Pâncreas/química , Hipófise , Ratos , Ratos Long-Evans , Ratos Sprague-Dawley , Ratos Transgênicos , Ratos Wistar , Análise de Regressão , Reprodutibilidade dos Testes , Serina/sangue , Serina/química , Serina/urina , Estereoisomerismo
19.
J Proteome Res ; 9(12): 6405-16, 2010 Dec 03.
Artigo em Inglês | MEDLINE | ID: mdl-20932058

RESUMO

Physical exercise modifies animal metabolism profoundly. Until recently, biochemical investigations related to exercise focused on a small number of biomolecules. In the present study, we used a holistic analytical approach to investigate changes in the human urine metabolome elicited by two exercise sessions differing in the duration of the rest interval between repeated efforts. Twelve men performed three sets of two 80 m maximal runs separated by either 10 s or 1 min of rest. Analysis of pre- and postexercise urine samples by (1)H NMR spectroscopy and subsequent multivariate statistical analysis revealed alterations in the levels of 22 metabolites. Urine samples were safely classified according to exercise protocol even when applying unsupervised methods of statistical analysis. Separation of pre- from postexercise samples was mainly due to lactate, pyruvate, hypoxanthine, compounds of the Krebs cycle, amino acids, and products of branched-chain amino acid (BCAA) catabolism. Separation of the two rest intervals was mainly due to lactate, pyruvate, alanine, compounds of the Krebs cycle, and 2-oxoacids of BCAA, all of which increased more with the shorter interval. Metabonomics provides a powerful methodology to gain insight in metabolic changes induced by specific training protocols and may thus advance our knowledge of exercise biochemistry.


Assuntos
Exercício Físico/fisiologia , Espectroscopia de Ressonância Magnética/métodos , Metaboloma/fisiologia , Metabolômica/métodos , Alanina/urina , Ácido Cítrico/urina , Creatinina/urina , Formiatos/urina , Fumaratos/urina , Histidina/urina , Humanos , Hidroxibutiratos/urina , Hipoxantina/urina , Cetoácidos/urina , Ácido Láctico/urina , Masculino , Análise Multivariada , Adulto Jovem
20.
J Am Soc Mass Spectrom ; 21(7): 1129-32, 2010 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-20304672

RESUMO

Sarcosine, an isomer of L-alanine, has been proposed as a prostate cancer progression biomarker [1]. Both compounds are detected in urine, where the measured sarcosine/alanine ratio has been found to be higher in prostate biopsy-positive group versus controls. We present here preliminary evidence showing that urine samples spiked with sarcosine/alanine can be partially resolved in 3 min via tandem differential mobility analysis-mass spectrometry (DMA-MS). Based on the calibration curves obtained for two mobility peaks, we finally estimate their concentration ratio in urine.


Assuntos
Alanina , Sarcosina , Espectrometria de Massas por Ionização por Electrospray/métodos , Espectrometria de Massas em Tandem/métodos , Alanina/isolamento & purificação , Alanina/urina , Biomarcadores Tumorais/química , Biomarcadores Tumorais/urina , Humanos , Isomerismo , Sarcosina/isolamento & purificação , Sarcosina/urina
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