RESUMO
We frequently encounter brownish-red, cloudy urine in some obese subjects, which occurs due to pink urine syndrome (PUS). PUS is a phenomenon in which uric acid precipitates into the urine due to reduced urinary pH (UpH). The mechanism underlying urinary acidification has not been elucidated so far. UpH level is adjusted by urinary excretion of ammonia synthesized from glutamate or glutamine, suggesting that renal synthesis of ammonia from glutamate or glutamine is decreased in PUS. However, this hypothesis has not been examined yet. We therefore examined the changes in the urinary excretion of these amino acids in PUS. One-hundred-fifty male students who had undergone a physical examination were enrolled. To determine the presence [PUS (+), n = 72] or absence [PUS (-), n = 78] of PUS, urinary amino acid excretion and UpH were evaluated. Independent risk factors of lower UpH were determined using multiple regression analyses. The PUS (+) subjects, who had lower UpH values than PUS (-) subjects, showed lower urinary excretion of glutamate and some other glucogenic amino acids. Thus, UpH correlated positively with the urinary excretion of glutamate in the PUS (+) subjects. A reduction in urinary glutamate but not in glutamine excretion proved to be an independent risk factor for reduced UpH. In conclusion, PUS appears to occur when a reduction in the synthesis of ammonia from glutamate causes a decrease in UpH. Our results showed that urinary glutamate excretion was reduced in PUS because renal glutamate was consumed by a reaction different from ammonia production.
Assuntos
Ácido Glutâmico/urina , Urina/química , Aminoácidos/metabolismo , Feminino , Glucose/metabolismo , Humanos , Concentração de Íons de Hidrogênio , Túbulos Renais Proximais/metabolismo , Túbulos Renais Proximais/patologia , Masculino , SíndromeRESUMO
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 EspecificidadeRESUMO
[14C]-Labeled arruva [sodium/potassium (2R,4R)-2-amino-4-carboxy-4-hydroxy-5-(3-indolyl) pentanoate] was administered as a single gavage dose (10 mg/kg bw) to male and female Beagle dogs and 1 bile duct-cannulated male. The mean peak arruva plasma concentration equivalent of 1.2 µg/g occurred at first sampling time point of 1 hour postdosing. The mean area under the concentration versus time curve from 0 hour postdosing to the last time point was approximately 20 µg·h/g and the mean terminal plasma elimination half-life ranged from 15 hours in females to 21 hours in males. Over 168 hours postdosing, 35% to 50% of the administered arruva was eliminated in the urine with 44% to 53% eliminated in feces; 1.3% of the administered dose was recovered in bile. Arruva and its derivatives were identified using tandem mass spectrometry, and the relative percentage of each substance was quantified via radio high-performance liquid chromatography. Over a 168-hour collection period, combined urine and feces extract data from the 6 noncannulated dogs showed that approximately 91% of the dose was excreted as unchanged parent arruva (41% in urine and 50% in feces). In the cannulated male, 95.3% was excreted as unchanged parent arruva; 50.2% in urine, 43.9% in feces, and 1.3% in bile. Lactone and lactam derivatives of arruva and 1 unidentified substance were detected in urine only during the first 24 hours postdosing with the greatest amounts detected during the first 6 hours of collection; up to 1% of lactone or lactam derivatives were detected in bile samples. Plasma pharmacokinetics data indicated rapid absorption of arruva with the majority of radioactivity located in the feces collected in the first 48 hours.
Assuntos
Ácido Glutâmico/análogos & derivados , Indóis/metabolismo , Absorção Intestinal , Adoçantes não Calóricos/metabolismo , Animais , Animais Endogâmicos , Bile/química , Radioisótopos de Carbono , Cromatografia Líquida de Alta Pressão , Cães , Fezes/química , Feminino , Ácido Glutâmico/sangue , Ácido Glutâmico/química , Ácido Glutâmico/metabolismo , Ácido Glutâmico/urina , Meia-Vida , Indóis/sangue , Indóis/química , Indóis/urina , Eliminação Intestinal , Cinética , Lactamas/sangue , Lactamas/química , Lactamas/metabolismo , Lactamas/urina , Lactonas/sangue , Lactonas/química , Lactonas/metabolismo , Lactonas/urina , Masculino , Estrutura Molecular , Adoçantes não Calóricos/química , Eliminação Renal , Caracteres Sexuais , Espectrometria de Massas em TandemRESUMO
BACKGROUND AND PURPOSE: Recent evidences indicate that glutamatergic homeostasis disorders are implicated in the pathogenesis of migraine. In particular, plasma and cerebrospinal fluid glutamate levels seem to be altered in migraine patients. However, the impacts of glutamate on migraine and especially on aura symptoms, alterations in the frequency of migraine attacks as well as investigations on glutamate on migraine-related metabolic dysfunctions, like hyperinsulinaemia, and an atherogenic lipid profile remain elusive to date. The aim of the present study was to investigate the impact of glutamate on migraine and related metabolic dysfunctions. METHODS: We investigated the urinary glutamate levels of female migraineurs (n = 48) in the interictal phase and healthy controls (n = 48). Parameters of the insulin- and lipid metabolism, inflammatory parameters and anthropometric parameters were additionally determined. RESULTS: Urinary glutamate levels of female migraineurs were significantly decreased with respect to the control group. Logistic regression revealed an odds ratio of 4.04 for migraine. We found a significant correlation with the time-period of patients' last attack and a significant inverse correlation with the annual frequency of migraine attacks. Other parameters of the insulin- and lipid metabolism, anthropometric and inflammatory parameters showed no significant correlation with glutamate levels. CONCLUSION: We show here that female migraineurs exhibit decreased urinary glutamate levels which are associated with a 4.04-fold higher risk for migraine and correlated with patients' frequency of migraine attacks.
Assuntos
Ácido Glutâmico/urina , Transtornos de Enxaqueca/urina , Adulto , Feminino , Humanos , Insulina/metabolismo , Metabolismo dos Lipídeos , Razão de ChancesRESUMO
In neonatal mammals, arginine is synthesized in the enterocyte, with either proline or glutamate as the dietary precursor. We have shown several times in piglets that proline is the only precursor to arginine, although in vitro evidence supports glutamate in this role. Because of this uncertainty, we performed a multitracer stable isotope study to determine whether proline, glutamate, or both are dietary precursors for arginine in enterally fed human neonates. Labeled arginine (M + 2), proline (M + 1), and glutamate (M + 3) were given enterally to 15 stable, growing preterm infants (GA at birth 30-35 wk) at 1-3 wk postnatal age. Enrichment in urine of the tracer amino acids and the M + 1 and M + 3 isotopomers of arginine were measured by LC-tandem mass spectrometry to determine the contribution of proline and glutamate to arginine synthesis. Plateau enrichments of arginine and proline tracers were measurable in urine. Urinary glutamate enrichment was not detected. Conversion of proline to arginine was detected. However, the M + 3 isotopomer of arginine, which would have been synthesized from glutamate, was not detected. We conclude that, in contrast to the current consensus in the literature based on in vitro studies, proline is the major contributor to arginine synthesis in human preterm infants.
Assuntos
Arginina/biossíntese , Nutrição Enteral , Recém-Nascido Prematuro/metabolismo , Prolina/metabolismo , Arginina/urina , Cromatografia Líquida , Citrulina/biossíntese , Ácido Glutâmico/metabolismo , Ácido Glutâmico/urina , Humanos , Recém-Nascido , Marcação por Isótopo , Ornitina/biossíntese , Prolina/urina , Espectrometria de Massas em TandemRESUMO
The number of people affected by Type 2 diabetes mellitus (T2DM) is close to half a billion and is on a sharp rise, representing a major and growing public health burden. Given its mild initial symptoms, T2DM is often diagnosed several years after its onset, leaving half of diabetic individuals undiagnosed. While several classical clinical and genetic biomarkers have been identified, improving early diagnosis by exploring other kinds of omics data remains crucial. In this study, we have combined longitudinal data from two population-based cohorts CoLaus and DESIR (comprising in total 493 incident cases vs. 1360 controls) to identify new or confirm previously implicated metabolomic biomarkers predicting T2DM incidence more than 5 years ahead of clinical diagnosis. Our longitudinal data have shown robust evidence for valine, leucine, carnitine and glutamic acid being predictive of future conversion to T2DM. We confirmed the causality of such association for leucine by 2-sample Mendelian randomisation (MR) based on independent data. Our MR approach further identified new metabolites potentially playing a causal role on T2D, including betaine, lysine and mannose. Interestingly, for valine and leucine a strong reverse causal effect was detected, indicating that the genetic predisposition to T2DM may trigger early changes of these metabolites, which appear well-before any clinical symptoms. In addition, our study revealed a reverse causal effect of metabolites such as glutamic acid and alanine. Collectively, these findings indicate that molecular traits linked to the genetic basis of T2DM may be particularly promising early biomarkers.
Assuntos
Carnitina/sangue , Diabetes Mellitus Tipo 2/diagnóstico , Predisposição Genética para Doença , Ácido Glutâmico/sangue , Leucina/sangue , Metaboloma/genética , Valina/sangue , Adulto , Idoso , Betaína/sangue , Betaína/urina , Biomarcadores/sangue , Biomarcadores/urina , Carnitina/urina , Estudos de Casos e Controles , Diabetes Mellitus Tipo 2/sangue , Diabetes Mellitus Tipo 2/genética , Diabetes Mellitus Tipo 2/urina , Diagnóstico Precoce , Feminino , Ácido Glutâmico/urina , Humanos , Leucina/urina , Lisina/sangue , Lisina/urina , Masculino , Manose/sangue , Manose/urina , Análise da Randomização Mendeliana , Pessoa de Meia-Idade , Valina/urinaRESUMO
Oxidative stress is suggested to play an important role in several pathophysiological conditions. A recent study showed that decreasing 5-oxoproline (pyroglutamate) concentration, an important mediator of oxidative stress, by over-expressing 5-oxoprolinase, improves cardiac function post-myocardial infarction in mice. The aim of the current study is to gain a better understanding of the role of the glutathione cycle in a mouse model of myocardial infarction by establishing quantitative relationships between key components of this cycle. We developed and validated an LC-MS method to quantify 5-oxoproline, L-glutamate, reduced glutathione (GSH) and oxidized GSH (GSSG) in different biological samples (heart, kidney, liver, plasma, and urine) of mice with and without myocardial infarction. 5-oxoproline concentration was elevated in all biological samples from mice with myocardial infarction. The ratio of GSH/GSSG was significantly decreased in cardiac tissue, but not in the other tissues/body fluids. This emphasizes the role of 5-oxoproline as an inducer of oxidative stress related to myocardial infarction and as a possible biomarker. An increase in the level of 5-oxoproline is associated with a decrease in the GSH/GSSG ratio, a well-established marker for oxidative stress, in cardiac tissue post-myocardial infarction. This suggests that 5-oxoproline may serve as an easily measurable marker for oxidative stress resulting from cardiac injury. Our findings show further that liver and kidneys have more capacity to cope with oxidative stress conditions in comparison to the heart, since the GSH/GSSG ratio is not affected in these organs despite a significant increase in 5-oxoproline.
Assuntos
Ácido Glutâmico/metabolismo , Glutationa/metabolismo , Rim/metabolismo , Fígado/metabolismo , Infarto do Miocárdio/metabolismo , Miocárdio/metabolismo , Ácido Pirrolidonocarboxílico/metabolismo , Animais , Cromatografia Líquida/métodos , Vasos Coronários/cirurgia , Ácido Glutâmico/sangue , Ácido Glutâmico/urina , Glutationa/sangue , Glutationa/urina , Ligadura , Camundongos , Oxirredução , Ácido Pirrolidonocarboxílico/sangue , Ácido Pirrolidonocarboxílico/urina , Espectrometria de Massas em Tandem/métodosRESUMO
The tyrosine, tryptophan, and glutamate metabolic pathways play key roles on pathological state of neuronal functions and the change of their levels in biological systems reflects the progress degree of neuronal diseases. Comprehensive profiling of these metabolites is important to find new biomarkers for diagnosis or prognosis of various neuronal diseases. However, the overall profiling analysis of various neurochemicals in biological sample is confronted with several limitations due to their low concentration and physicochemical properties and the coexistence of matrices. We developed an efficient and feasible method using gas chromatography-tandem mass spectrometry (GC-MS/MS). Wide-bore mixed cation exchange (MCX) SPE process enables a rapid and effective cleanup of 20 neurochemicals even including acidic and basic neurochemicals in a single SPE cartridge by using different composition of eluents. Selective derivatization of various types of metabolites was applied to achieve highly chromatographic separation and sensitive mass detection. Appropriate selection of precursor and product transition ions used in multiple reaction-monitoring (MRM) mode based on the MS/MS fragmentations of the derivatized neurochemicals could be significantly minimized the matrix effects and enhanced the reliability of quantification results. The developed method was validated in terms of linearity, limits of detection, precision, accuracy, and matrix effects. The intra- and inter-assay analytical variations were less than 10%. The overall linearity for all of the targets was excellent (R2≥0.996). The detection limits ranged between 0.38 and 8.13ng/mL for the acidic neurochemicals and between 0.02 and 11.1ng/mL for the basic neurochemicals. The developed protocol will be expected to be a promising tool for the understanding of the pathological state and diagnosis of various neuronal diseases.
Assuntos
Cromatografia Gasosa-Espectrometria de Massas/métodos , Ácido Glutâmico/urina , Metaboloma , Extração em Fase Sólida/métodos , Triptofano/urina , Tirosina/urina , Biomarcadores/metabolismo , Biomarcadores/urina , Ácido Glutâmico/metabolismo , Humanos , Limite de Detecção , Metabolômica/métodos , Espectrometria de Massas em Tandem/métodos , Triptofano/metabolismo , Tirosina/metabolismoRESUMO
AIM: To identify metabolic signatures in urine samples from healthy and inflammatory bowel disease (IBD) children. METHODS: We applied liquid chromatography and gas chromatography coupled to targeted mass spectrometry (MS)-based metabolite profiling to identify and quantify bile acids and host-gut microbial metabolites in urine samples collected from 21 pediatric IBD patients monitored three times over one year (baseline, 6 and 12 mo), and 27 age- and gender-matched healthy children. RESULTS: urinary metabolic profiles of IBD children differ significantly from healthy controls. Such metabolic differences encompass central energy metabolism, amino acids, bile acids and gut microbial metabolites. In particular, levels of pyroglutamic acid, glutamic acid, glycine and cysteine, were significantly higher in IBD children in the course of the study. This suggests that glutathione cannot be optimally synthesized and replenished. Whilst alterations of the enterohepatic circulation of bile acids in pediatric IBD patients is known, we show here that non-invasive urinary bile acid profiling can assess those altered hepatic and intestinal barrier dysfunctions. CONCLUSION: The present study shows how non-invasive sampling of urine followed by targeted MS-based metabonomic analysis can elucidate and monitor the metabolic status of children with different GI health/disease status.
Assuntos
Ácidos e Sais Biliares/urina , Microbioma Gastrointestinal , Doenças Inflamatórias Intestinais/microbiologia , Doenças Inflamatórias Intestinais/urina , Metaboloma , Urina/química , Adolescente , Antropometria , Composição Corporal , Estudos de Casos e Controles , Criança , Colite Ulcerativa/urina , Doença de Crohn/urina , Cisteína/urina , Feminino , Cromatografia Gasosa-Espectrometria de Massas , Ácido Glutâmico/urina , Glutationa/urina , Glicina/urina , Humanos , Inflamação , Masculino , Metabolômica , Interações Microbianas , Fenótipo , Ácido Pirrolidonocarboxílico/urina , Transdução de SinaisRESUMO
Male New Zealand White rabbits received a single intravenous injection of 125 mg/kg cephaloridine, 500 mg/kg cefoperazone or 1000 mg/kg cephalothin. Histological examination of kidneys at 48 h post-dose confirmed the presence of bilateral necrosis of the proximal convoluted tubules in the cephaloridine-treated animals. 1H-NMR urinalysis of cephaloridine-treated rabbits detected drug-related resonances, decreased hippurate and increased glucose at 0-24 h post-dose accompanied by elevated levels of lactate, glycine, citrate, glutamine/glutamate and alanine at 24-48 h post-dose. No histopathological changes were observed following administration of cefoperazone or cephalothin. 1H-NMR spectra of urine collected from these animals showed drug-related resonances and decreased hippurate levels at 0-24 h post-dose, and increased glucose levels at 24-48 h post-dose. Analysis of urine by conventional clinical-chemistry failed to reveal any statistically significant differences between the treatment groups. Under the conditions of this study, the nephrotoxic effects of cephaloridine and the minimal effects of cefoperazone and cephalothin could be clearly distinguished by 1H-NMR urinalysis but not by conventional urinalysis.
Assuntos
Cefalosporinas/toxicidade , Túbulos Renais Proximais/efeitos dos fármacos , Alanina/urina , Animais , Cefoperazona/administração & dosagem , Cefoperazona/toxicidade , Cefaloridina/administração & dosagem , Cefaloridina/toxicidade , Cefalosporinas/administração & dosagem , Cefalotina/administração & dosagem , Cefalotina/toxicidade , Citratos/urina , Ácido Cítrico , Relação Dose-Resposta a Droga , Ácido Glutâmico/urina , Glicina/urina , Glicosúria/urina , Hipuratos/urina , Injeções Intravenosas , Túbulos Renais Proximais/patologia , Lactatos/urina , Ácido Láctico , Espectroscopia de Ressonância Magnética , Masculino , Coelhos , Urinálise/normasRESUMO
We determined the urinary amino acid concentrations by high-performance liquid chromatography (HPLC) using an eluent buffer which had been processed through a column packed with strong cation-exchange resin. With this column, ghost peaks resolved or much reduced in size and shifted to a post-arginine position. A clearer separation of peaks of urinary amino acids and ammonia was obtained. This could be due to the elimination of O-phthalaldehyde reactive substances in the eluent by the resin column. This method does not require a separate step to remove ammonia, unlike most of the methods currently used, and thus saves time.
Assuntos
Aminoácidos/urina , Cromatografia Líquida de Alta Pressão/instrumentação , Cromatografia Líquida de Alta Pressão/métodos , Asparagina/urina , Ácido Aspártico/urina , Ácido Glutâmico/urina , Glutamina/urina , Humanos , Reprodutibilidade dos TestesRESUMO
AIMS: To investigate whether the duration of chronic pain in temporomandibular disorder (TMD) patients is associated with a net depletion of amino acids, and a distinct process from pain intensity. METHODS: Twenty-nine patients defined by the research diagnostic criteria/TMD as having Type 1a muscle pain (TMD1A group), and 34 age- and sex-matched control subjects, were assessed for variation in urinary organic and amino acid excretion by gas chromatography-mass spectrometry. RESULTS: The TMD1A patients' mean pain intensity, assessed on a visual analog scale (VAS), was 5.4 (95% confidence limits: 4.5 to 6.3), TMD1A illness duration was 5.0 +/- 1.2 (SD) years, number of body areas with pain/subject was 6.3 +/- 2.4 (range 0 to 10), and symptom prevalence from the Symptom Check List-90-Revised (SCL-90-R) was 25.5 +/- 11.3 symptoms/subject, which was higher than the controls (5.2 +/- 5.0 symptoms/subject, P < .001). TMD1A patient illness duration was positively correlated with symptom prevalence and body pain distribution, and all were independent of pain intensity. The TMD1A patients had: (1) and increased tyrosine:leucine ratio; and (2) reduced leucine concentrations (both P < .001), which suggests deregulated catabolism. Pain intensity was associated with: (1) changes in the multivariate urinary metabolite excretion patterns (P < .001); (2) reduced leucine concentrations (P < .001); and (3) increases in total urinary metabolites (P < .04), and in 2 unidentified molecules, UM28 (P < .001) and CFSUM1 (P < .002). TMD1A illness duration was associated with lower (1) urinary metabolite concentrations and (2) succinic acid and combined glutamine + glutamic acid levels, suggesting a progressive depletion of metabolite reserves. CONCLUSION: In TMD1A patients, total amino acid excretion was positively correlated with pain intensity and negatively correlated with illness duration, which indicated that illness duration was associated with a different set of metabolic anomalies compared with those identified for pain intensity.
Assuntos
Dor Facial/fisiopatologia , Medição da Dor , Desnaturação Proteica , Transtornos da Articulação Temporomandibular/fisiopatologia , Adulto , Aminoácidos/urina , Estudos de Casos e Controles , Distribuição de Qui-Quadrado , Doença Crônica , Estudos de Coortes , Intervalos de Confiança , Dor Facial/metabolismo , Feminino , Cromatografia Gasosa-Espectrometria de Massas , Ácido Glutâmico/urina , Glutamina/urina , Humanos , Leucina/urina , Masculino , Ácido Succínico/urina , Transtornos da Articulação Temporomandibular/metabolismo , Fatores de Tempo , Tirosina/urinaRESUMO
AIMS: To investigate the association between toxin-producing staphylococci, symptom expression, and changes in urinary excretion of metabolites in temporomandibular disorder (TMD) patients and age- and sex-matched control subjects. METHODS: Twenty-nine patients defined by the research diagnostic criteria/TMD as having Type 1a muscle pain (TMD1A), and 34 age- and sex-matched control subjects were assessed for the carriage of staphylococcal species, staphylococcal toxin production, expression of symptoms, and changes in urinary excretion of amino and organic acids. RESULTS: TMD1A patients had an increased incidence of carriage of toxin-producing coagulase-negative staphylococcus (MDT-CoNS, P < .004), which produced increased levels of delta-like membrane-damaging toxins. The TMD1A patients also had a reduction in the incidence of carriage of Staphylococcus aureus (P < .02). Increased incidence of MDT-CoNS was positively associated with increased pain intensity as assessed by a visual analog scale (P < .001). Odds ratio analysis revealed a 9.2-fold increase in MDT-CoNS recovery from the nose of TMD1A patients compared with the control subjects (odds ratio = 9.2, > 95% confidence limits: 2.3 to 37.5, P < .001). Increases in the carriage incidence of MDT-CoNS were also associated with increases in the urinary tyrosine:leucine ratio (P < .004), which represents a change in the balance of proteolysis and protein synthesis. The toxin production by these CoNS species was also associated with an increased urinary excretion of glutamic acid (P < .03). CONCLUSION: These data suggest that an increased colonization of MDT-CoNS on skin and mucosal membranes was associated with changed proteolysis, increased pain intensity, and an increase in excitatory amino acids consistent with events associated with the development of chronic orofacial muscle pain in TMD patients.
Assuntos
Toxinas Bacterianas/análise , Dor Facial/microbiologia , Medição da Dor , Staphylococcus/classificação , Transtornos da Articulação Temporomandibular/microbiologia , Adulto , Aminoácidos/urina , Estudos de Casos e Controles , Doença Crônica , Intervalos de Confiança , Dor Facial/fisiopatologia , Dor Facial/urina , Feminino , Ácido Glutâmico/urina , Proteínas Hemolisinas/análise , Humanos , Leucina/urina , Masculino , Mucosa Nasal/microbiologia , Razão de Chances , Staphylococcus aureus/isolamento & purificação , Transtornos da Articulação Temporomandibular/fisiopatologia , Transtornos da Articulação Temporomandibular/urina , Tirosina/urinaRESUMO
There are many reports about the significant roles of some amino acids in neurobiology and treatment of autism. This is a critical review of amino acids levels in autism. No published review article about the level of amino acids in autism was found. The levels of glutamate and homocystein are increased in autism while the levels of glutamine and tryptophan are decreased. Findings regarding the plasma levels of taurine and lysine are controversial. The urinary levels of homocysteine and essential amino acids in both the untreated and treated autistic children are significantly less than those in the controls. The current literature suffers from many methodological shortcomings which needed to be considered in future studies. Some of them are age, gender, developmental level, autism symptoms severity, type of autism spectrum disorders, medical comorbidities, intelligent quotient, diet, concomitant medications, body mass index, and technical method of assessment of amino acids.
Assuntos
Transtorno Autístico/sangue , Ácido Glutâmico/sangue , Homocisteína/sangue , Taurina/sangue , Transtorno Autístico/diagnóstico , Biomarcadores/sangue , Biomarcadores/urina , Ácido Glutâmico/urina , Homocisteína/urina , Humanos , Taurina/urinaRESUMO
For a metabolomics study focusing on the analysis of aspartic and glutamic acid enantiomers, a fully automated two-dimensional HPLC system employing a microbore-ODS column and a narrowbore-enantioselective column was developed. By using this system, a detailed distribution of D-Asp and D-Glu besides L-Asp and L-Glu in mammals was elucidated. For the total analysis concept, the amino acids were first pre-column derivatized with 4-fluoro-7-nitro-2,1,3-benzoxadiazole (NBD-F) to be sensitively and fluorometrically detected. For the non-stereoselective separation of the analytes in the first dimension a monolithic ODS column (750 mm × 0.53 mm i.d.) was adopted, and a self-packed narrowbore-Pirkle type enantioselective column (Sumichiral OA-2500S, 250 mm × 1.5 mm i.d.) was selected for the second dimension. In the rat plasma, RSD values for intra-day and inter-day precision were less than 6.8%, and the accuracy ranged between 96.1% and 105.8%. The values of LOQ of D-Asp and D-Glu were 5 fmol/injection (0.625 nmol/g tissue). The present method was successfully applied to the simultaneous determination of free aspartic acid and glutamic acid enantiomers in 7 brain areas, 11 peripheral tissues, plasma and urine of Wistar rats. Biologically significant D-Asp values were found in various tissue samples whereas for D-Glu the values were very low possibly indicating less significance.
Assuntos
Cromatografia Líquida de Alta Pressão/métodos , Ácido D-Aspártico/análise , Ácido Glutâmico/análise , 4-Cloro-7-nitrobenzofurazano/análogos & derivados , 4-Cloro-7-nitrobenzofurazano/química , Animais , Química Encefálica , Ácido D-Aspártico/sangue , Ácido D-Aspártico/química , Ácido D-Aspártico/urina , Ácido Glutâmico/sangue , Ácido Glutâmico/química , Ácido Glutâmico/urina , Modelos Lineares , Masculino , Especificidade de Órgãos , Ratos , Ratos Wistar , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , EstereoisomerismoRESUMO
Previously published studies have shown D-glutamate to be the most potent natural inhibitor of glutathione synthesis known, yet how D-glutamate is handled in humans is unknown. Therefore, we administered an oral D-glutamate load to four healthy volunteers and monitored the plasma D-glutamate concentration and excretion over a 3-h postload period. Compared with time controls, the plasma D-glutamate concentration increased 10-fold in the 1st h and then reached a plateau over the remaining time course. In contrast, plasma D-pyrrolidone carboxylic acid increased progressively throughout the 3-h time course to a level 10-fold higher than the D-glutamate plasma concentration. Excretion of D-glutamate progressively increased despite a constant filtered D-glutamate load rising from only 5 to 95% of the filtered amount. Excretion of D-pyrrolidone carboxylic acid increased with the rise in filtered load without significant reabsorption. The amount of D-pyrrolidone carboxylic acid excreted over the 3-h time course was 10 times the amount excreted as D-glutamate and accounted for almost 20% of the administered D-glutamate. These findings indicate that plasma D-glutamate concentration is tightly regulated through two mechanisms: 1) the transport into cells and metabolic conversion to D-pyrrolidone carboxylic acid and excretion, and 2) the enhancement of D-glutamate clearance by the kidneys.
Assuntos
Ácido Glutâmico/farmacocinética , Ácido Pirrolidonocarboxílico/metabolismo , Absorção , Administração Oral , Adulto , Ácido Glutâmico/sangue , Ácido Glutâmico/urina , Humanos , Masculino , Ácido Pirrolidonocarboxílico/sangue , Ácido Pirrolidonocarboxílico/urina , Fatores de TempoRESUMO
Assays of the branched chain amino acids, of those with basic character, of phenylalanine and glutamic acid were performed in the 24 hrs urines of 50 patients with psychic disorders, ranging in age from 0 to 20 years, and in 30 normal controls. Similar investigations were carried out in the patients' parents. Abnormal levels of urinary amino acids in some of the adults suggested the existence of mutant genes in one or both parents of one or two descendents with mental handicap. The levels of urinary amino acids correlated with the severity of the predominant psychic manifestations, which were more severe in the cases with a wide deviation from normal of aminoaciduria.
Assuntos
Erros Inatos do Metabolismo dos Aminoácidos/urina , Aminoácidos de Cadeia Ramificada/urina , Aminoácidos Essenciais/urina , Transtornos Mentais/urina , Adolescente , Adulto , Erros Inatos do Metabolismo dos Aminoácidos/genética , Criança , Pré-Escolar , Feminino , Ácido Glutâmico/urina , Humanos , Lactente , Masculino , Transtornos Mentais/genética , Fenilalanina/urinaRESUMO
Homozygous NEUT2 mice lack cytosolic 10-formyltetrahydrofolate dehydrogenase (FDH; Champion et al. (1994) Proc. Natl. Acad. Sci. USA 91, 11,338-11,342) and as a consequence should be unable to oxidize carbon 2 of l-histidine to CO2 via 10-formyltetrahydrofolate in liver cytosol. There was essentially no oxidation of l-[2-14C]histidine to 14CO2 in homozygous NEUT2 mice, but 52% of the [2-14C]l-histidine dose was recovered in the urine within 24 h. Analysis of urine samples for [14C]formiminoglutamate, the expected excretion product, was negative; however, [14C]urocanic acid was detected. Investigation of histidine catabolism via the folate-dependent deamination pathway revealed no detectable urocanase activity in homozygous NEUT2 mice, while heterozygous NEUT2 mice had 50% urocanase activity compared to normal mice. Histidase and formiminotransferase-cyclodeaminase, also on the histidine deamination pathway, had similar specific activities in normal and NEUT2 mice. Histidine-pyruvate aminotransferase, the first enzyme of the alternate histidine transamination catabolic pathway did not appear to be affected by the loss of urocanase. Based on the excretion of urocanic acid it is estimated that NEUT2 mice catabolize approximately 40 micromol/day via the deamination pathway. The loss of urocanase activity in homozygous NEUT2 mice may allow these mice to survive the disruption in folate metabolism by sparing the liver cytosolic tetrahydrofolate pool.
Assuntos
Histidina/metabolismo , Fígado/enzimologia , Animais , Citosol/metabolismo , Genótipo , Ácido Glutâmico/urina , Heterozigoto , Homozigoto , Fígado/metabolismo , Camundongos , Camundongos Knockout , Modelos Químicos , Oxigênio/metabolismo , Transaminases/metabolismo , Ácido Urocânico/urinaRESUMO
Chronic fatigue syndrome (CFS/ME) is a debilitating fatigue illness that has an unknown etiology. We studied 20 chronic fatigue syndrome (CFS) patients, who complied with the Oxford and American CDC definitions, and 45 non-CFS subjects. Participants completed questionnaires, were clinically examined, and had first morning urine specimens collected, which were screened by gas chromatography-mass spectrometry for changes in metabolite excretion. Multivariate analysis of the urinary metabolite profiles differed significantly in the CFS patients compared to the non-CFS patients (P < 0.004). The CFS patients had increases in aminohydroxy-N-methylpyrrolidine (P < 0.00003, referred to as chronic fatigue symptom urinary marker 1, or CFSUM1), tyrosine (P < 0.02), beta-alanine (P < 0.02), aconitic acid (P < 0.05), and succinic acid (P < 0.05) and reductions in an unidentified urinary metabolite, CFSUM2 (P < 0.0007), alanine (P < 0.005), and glutamic acid (P < 0.02). CFSUM1, beta-alanine, and CFSUM2 were found by discriminant function analysis to be the first, second, and third most important metabolites, respectively for discriminating between CFS and non-CFS subjects. The abundances of CFSUM1 and beta-alanine were positively correlated with symptom incidence (P < 0.01 and P < 0.001, respectively), symptom severity, core CFS symptoms, and SCL-90-R somatization (P < 0.00001), suggesting a molecular basis for CFS.
Assuntos
Síndrome de Fadiga Crônica/urina , Ácido Aconítico/urina , Adolescente , Adulto , Alanina/urina , Aminoácidos/urina , Análise de Variância , Biomarcadores/urina , Síndrome de Fadiga Crônica/fisiopatologia , Síndrome de Fadiga Crônica/psicologia , Feminino , Cromatografia Gasosa-Espectrometria de Massas , Ácido Glutâmico/urina , Humanos , Masculino , Pessoa de Meia-Idade , Análise Multivariada , Exame Físico , Pirrolidinas/urina , Valores de Referência , Succinatos/urina , Ácido Succínico , Inquéritos e Questionários , Tirosina/urina , beta-Alanina/urinaRESUMO
OBJECTIVES: Animal studies have shown that the elevation of plasma glutamate levels increase cerebral edema formation whenever the blood-brain barrier is disturbed. Therefore, changes in plasma glutamate levels as influenced by the administration of a glutamate-containing amino acid solution were investigated in neurosurgical patients. DESIGN: Prospective, descriptive study. SETTING: Eight-bed neurosurgical intensive care unit in a university hospital. PATIENTS: Twenty-three neurosurgical patients requiring parenteral nutrition. INTERVENTIONS: Parenteral nutrition was begun 24 hrs after craniotomy. Patients receiving a glutamate-containing amino acid solution (3.75 g/L glutamate) were compared with patients infused with a glutamate-free solution. MEASUREMENTS AND MAIN RESULTS: Arterial plasma and urine amino acids were analyzed using high-performance liquid chromatography. Administration of a glutamate-containing solution doubled plasma glutamate levels in neurosurgical patients (from 53.3 +/- 9.8 microM [preinfusion] to 98.5 +/- 18.7 microM [after 4 hrs of infusion]; p < 0.001), whereas no elevation was seen when infusing a glutamate-free solution (from 52.3 +/- 7.3 [1 hr of infusion] to 53.6 +/- 6.4 microM [4 hrs of infusion]). Upon terminating the glutamate-containing infusion, arterial plasma glutamate levels decreased immediately (from 120 +/- 13.2 microM to 81.2 +/- 19.5 microM). Glutamate as infused in excess appears to exceed a renal threshold and is eliminated renally. CONCLUSIONS: As shown in animal models, administration of a glutamate-containing amino acid solution significantly increased plasma glutamate levels. Because such an increase in plasma glutamate levels could aggravate cerebral edema formation, glutamate-containing amino acid solutions cannot be recommended for patients with a disturbed blood-brain barrier.