RESUMO
OBJECTIVE: High fructose consumption exacerbates purine degradation and intestinal dysbiosis, which are closely related to the development of hyperuricemia. Probiotics are powerful weapons to combat metabolic disturbance and intestinal dysbiosis. Previously we isolated a Lactobacillus strain named DM9218 that could reduce the serum uric acid (UA) level by assimilating purine nucleosides. The present study aimed to evaluate the effects of DM9218 on high-fructose-induced hyperuricemia and to elucidate the underlying mechanisms. METHODS: Mice were fed a normal diet, a high-fructose diet, or high-fructose diet with DM9218. Metabolic parameters, fructose- and UA-related metabolites, and fecal microbiota were investigated. Whole-genome sequencing of strain DM9218 was also conducted. In addition, an inosine hydrolase from DM9218 was heterologously expressed in Escherichia coli, and its inosine-degrading activity was detected. RESULTS: Our results indicated that DM9218 could decrease serum UA level and hepatic xanthine oxidase activity in fructose-fed mice. It could protect against high-fructose-induced liver damage and retard UA accumulation by degrading inosine. The modulation effect of DM9218 on high-fructose-induced intestinal dysbiosis resulted in enhancement of intestinal barrier function and reduction of liver lipopolysaccharide, which was closely correlated with the down-regulation of inflammatory cytokine-stimulated xanthine oxidase expression and activity. CONCLUSIONS: Lactobacillus brevis DM9218 is a probiotic strain with the potential to ameliorate fructose-induced hyperuricemia.
Assuntos
Disbiose/tratamento farmacológico , Frutose/administração & dosagem , Microbioma Gastrointestinal/efeitos dos fármacos , Hiperuricemia/tratamento farmacológico , Inosina/metabolismo , Levilactobacillus brevis , Animais , Dieta/efeitos adversos , Dieta/métodos , Modelos Animais de Doenças , Disbiose/metabolismo , Disbiose/urina , Hiperuricemia/etiologia , Hiperuricemia/urina , Inosina/urina , Intestinos/efeitos dos fármacos , Intestinos/microbiologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , ProbióticosRESUMO
Cystinuria is an autosomal recessive disease that causes L-cystine precipitation in urine and nephrolithiasis. Disease severity is highly variable; it is known, however, that cystinuria has a more severe course in males. The aim of this study was to compare L-cystine metastability in first-morning urine collected from 24 normal female and 24 normal male subjects. Samples were buffered at pH 5 and loaded with L-cystine (0.4 and 4 mM final concentration) to calculate the amount remaining in solution after overnight incubation at 4 °C; results were expressed as Z scores reflecting the L-cystine solubility in each sample. In addition, metabolomic analyses were performed to identify candidate compounds that influence L-cystine solubility. L-cystine solubility Z score was +0.44 ± 1.1 and -0.44 ± 0.70 in female and male samples, respectively (p < 0.001). Further analyses showed that the L-cystine solubility was independent from urine concentration but was significantly associated with low urinary excretion of inosine (p = 0.010), vanillylmandelic acid (VMA) (p = 0.015), adenosine (p = 0.029), and guanosine (p = 0.032). In vitro L-cystine precipitation assays confirmed that these molecules induce higher rates of L-cystine precipitation in comparison with their corresponding dideoxy molecules, used as controls. In silico computational and modeling analyses confirmed higher binding energy of these compounds. These data indicate that urinary excretion of nucleosides and VMA may represent important factors that modulate L-cystine solubility and may represent new targets for therapy in cystinuria.
Assuntos
Cisteína/urina , Adenosina/urina , Adulto , Precipitação Química , Cisteína/química , Cistinúria/urina , Feminino , Guanosina/urina , Humanos , Inosina/urina , Masculino , Pessoa de Meia-Idade , Caracteres Sexuais , Solubilidade , Ácido Vanilmandélico/urinaRESUMO
Quantitative nontarget analysis is intended to provide a measurement of concentration of newly identified components in complex biological or environmental samples for which authentic or labeled standard do not exist. Electrospray ionization-high field asymmetric waveform ion mobility spectrometry-mass spectrometry (ESI-FAIMS-MS) has unique advantages that allowed us to develop a novel approach for quantification of nontarget analytes. In the nontarget analysis of urinary metabolites by ESI-FAIMS-MS, we find it practical and beneficial to analyze highly diluted urine samples. We show that urine extracts can be analyzed directly at very high dilutions (up to 20,000 times) by extending MS analysis times during slow FAIMS scanning. We explore the effects of sample dilution on ionization efficiency and ionization suppression in direct electrospray of complex sample matrixes. We consistently observe two distinct regimes in ESI operation related to the limited ionization capacity of this method. In the linear dynamic concentration range below the limiting ionization capacity, the analytical sensitivity of an analyte is constant and does not depend on matrix composition and concentration. Once the capacity of ESI is exceeded, all species exhibit log-log linearity in signal response. We show how quantification can be carried out using two different approaches, one for analytes which can be detected in the linear regime and another for those only detected in the suppression regime that overcomes the effects of ionization suppression. Our new insight into ionization suppression effects in ESI is of broad interest to anyone using ESI as an ionization technique for the MS analysis of complex samples.
Assuntos
Espectrometria de Massas por Ionização por Electrospray/métodos , Guanosina/análogos & derivados , Guanosina/metabolismo , Guanosina/urina , Inosina/análogos & derivados , Inosina/metabolismo , Inosina/urina , Íons/química , Espectrometria de Massas por Ionização por Electrospray/instrumentação , UrináliseRESUMO
This article describes the selective determination of inosine (INO) in the presence of important physiological interferents, uric acid (UA) and hypoxanthine (HXN), by differential pulse voltammetry at physiological pH (7.2) using the electropolymerized film of 3-amino-5-mercapto-1,2,4-triazole (p-AMTa) modified glassy carbon (GC) electrode. The electropolymerization of AMTa was carried out by the potentiodynamic method in 0.1M H(2)SO(4). An atomic force microscopy image shows that the p-AMTa film contains a spherical-like structure. Bare GC electrode fails to resolve the voltammetric signal of INO in the presence of UA and HXN due to the surface fouling caused by the oxidized products of UA and HXN. However, p-AMTa film modified GC electrode (p-AMTa electrode) not only separates the voltammetric signals of UA, HXN, and INO, with potential differences of 730 mV between UA and HXN and 310 mV between HXN and INO, but also shows enhanced oxidation current for them. The selective determination of INO in the presence of UA and HXN at physiological pH was achieved for the first time. Using the amperometric method, we achieved the lowest detection of 50 nM for INO. The practical application of the current modified electrode was demonstrated by determining the concentration of INO in human blood serum and urine samples.
Assuntos
Técnicas Biossensoriais/métodos , Hipoxantina/análise , Inosina/análise , Ácido Úrico/análise , Técnicas Eletroquímicas , Eletrodos , Humanos , Concentração de Íons de Hidrogênio , Hipoxantina/sangue , Hipoxantina/urina , Inosina/sangue , Inosina/urina , Microscopia de Força Atômica , Oxirredução , Triazóis , Ácido Úrico/sangue , Ácido Úrico/urinaRESUMO
Voltammetric determination of adenosine and inosine has been carried out at single-wall carbon nanotubes (SWNTs) modified pyrolytic graphite electrode (PGE) at pH 7.2 using Osteryoung square wave voltammetry (OSWV). The modified electrode exhibits remarkable electrocatalytic properties towards adenosine and inosine oxidation with a peak potential of approximately 1229 mV and 1348 mV, respectively. Linear calibration curves are obtained over the concentration range 0.5 microM to 1.0 mM in adenosine and 10 microM to 1.0 mM in inosine with sensitivity of 1.0 microA microM(-1) and 1.9 microA microM(-1) for adenosine and inosine respectively. The limit of detection for adenosine and inosine was found to be 0.51x10(-7) M and 2.04x10(-7) M, respectively. The proposed method was also used to estimate these compounds in human blood plasma and urine samples and the method was validated using HPLC.
Assuntos
Adenosina/análise , Eletrodos/normas , Inosina/análise , Nanotubos de Carbono , Adenosina/sangue , Adenosina/urina , Grafite , Inosina/sangue , Inosina/urina , Métodos , OxirreduçãoRESUMO
Modified nucleosides are formed post-transcriptionally in RNA. In cancer disease, the cell turnover and thus RNA metabolism is increased, yielding higher concentrations of excreted modified nucleosides. In the presented study, urinary ribonucleosides were used to differentiate between breast cancer patients and healthy volunteers. The nucleosides were extracted from urine samples using affinity chromatography and subsequently analyzed via liquid chromatography ion trap mass spectrometry (LC-IT-MS). The peak areas were related to the internal standard isoguanosine and to the urinary creatinine level. For bioinformatic pattern recognition we used the support vector machine. We examined 113 urine samples from breast cancer patients (stage Tis-T4) and 99 control samples from healthy volunteers. We achieved a sensitivity of 87.67% and a specificity of 89.90% when including 31 nucleosides. The medical metabonomics concept based on the urinary nucleoside profile reveals a significantly improved classification compared with currently applied breast cancer biomarkers such as CA15-3.
Assuntos
Neoplasias da Mama/diagnóstico , Espectrometria de Massas/métodos , Nucleosídeos/urina , Adenosina/urina , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Algoritmos , Inteligência Artificial , Biomarcadores/urina , Neoplasias da Mama/urina , Citidina/urina , Reações Falso-Negativas , Reações Falso-Positivas , Feminino , Guanosina/urina , Humanos , Inosina/urina , Pessoa de Meia-Idade , Análise de Componente Principal , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Uridina/urinaRESUMO
Stimulation of adenylyl cyclase causes cellular efflux of cAMP, and cAMP (unlike adenosine) is stable in blood. Therefore, it is conceivable that cAMP could function as a circulating adenosine prohormone by local target-organ conversion of distally released cAMP to adenosine via the sequential actions of ectophosphodiesterase and ecto-5'-nucleotidase (cAMP==> AMP==> adenosine; called the cAMP-adenosine pathway). A possible specific representation of this general concept is the pancreatohepatorenal cAMP-adenosine mechanism. The pancreas secretes glucagon into the portal circulation, and glucagon is a stimulant of hepatic adenylyl cyclase. Therefore, we hypothesize that the pancreas, via glucagon, stimulates hepatic cAMP production, which provides circulating cAMP for conversion to adenosine in the kidney via the cAMP-adenosine pathway. In normal rats, intravenous cAMP increased urinary and renal interstitial (assessed by renal microdialysis) cAMP and adenosine. Intraportal infusions of glucagon increased plasma cAMP 10-fold, it did not affect plasma adenosine, and it increased urinary and renal interstitial cAMP and adenosine. Local renal interstitial blockade (by adding inhibitors directly to the microdialysis perfusate) of ectophosphodiesterase (using 3-isobutyl-1-methylxanthine or 1,3-dipropyl-8-p-sulfophenylxanthine) or ecto-5'-nucleotidase (using alpha,beta-methyleneadenosine-5'-diphosphate) prevented the cAMP-induced and glucagon-induced increases in renal interstitial adenosine, but not cAMP. In ZSF1 rats with the metabolic syndrome, an oral glucose load increased plasma glucagon and urinary cAMP and adenosine excretion. We conclude that circulating cAMP is a substrate for local conversion to adenosine via the cAMP-adenosine pathway. A specific manifestation of this is the pancreatohepatorenal cAMP-adenosine mechanism (pancreas==> portal glucagon==> liver==> circulating cAMP==> kidney==> local cAMP-adenosine pathway).
Assuntos
Adenosina/urina , AMP Cíclico/metabolismo , Rim/metabolismo , Fígado/metabolismo , Pâncreas/fisiologia , Monofosfato de Adenosina/urina , Animais , AMP Cíclico/administração & dosagem , Glucagon/farmacologia , Glucose/farmacologia , Inosina/urina , Masculino , Microdiálise , Probenecid/farmacologia , RatosRESUMO
The influence of sprint training on endogenous urinary purine loss was examined in 7 active male subjects (age, 23.1 +/- 1.8 y; body mass, 76.1 +/- 3.1 kg; VO2 peak, 56.3 +/- 4.0 mL.kg-1.min-1). Each subject performed a 30 s sprint performance test (PT), before and after 7 d of sprint training. Training consisted of 15 sprints, each lasting 10 s, on an air-braked cycle ergometer performed twice each day. A rest period of 50 s separated each sprint during training. Sprint training resulted in a 20% higher muscle ATP immediately after PT, a lower IMP (57% and 89%, immediately after and 10 min after PT, respectively), and inosine accumulation (53% and 56%, immediately after and 10 min after the PT, respectively). Sprint training also attenuated the exercise-induced increases in plasma inosine, hypoxanthine (Hx), and uric acid during the first 120 min of recovery and reduced the total urinary excretion of purines (inosine + Hx + uric acid) in the 24 h recovery period following intense exercise. These results show that intermittent sprint training reduces the total urinary purine excretion after a 30 s sprint bout.
Assuntos
Educação Física e Treinamento , Purinas/urina , Corrida , Adulto , Humanos , Hipoxantina/sangue , Hipoxantina/urina , Inosina/sangue , Inosina/urina , Masculino , Músculos/metabolismo , Concentração Osmolar , Fatores de Tempo , Ácido Úrico/sangue , Ácido Úrico/urinaRESUMO
The influence of allopurinol on urinary purine loss was examined in 7 active male subjects (age 24.9 +/- 3.0 years, weight 82.8 +/- 8.3 kg, V O2peak 48.1 +/- 6.9 mL.kg(-1).min(-1)). These subjects performed, in random order, a trial with 5 days of prior ingestion of a placebo or allopurinol. Each trial consisted of eight 10-second sprints on an air-braked cycle ergometer and was separated by at least a week. A rest period of 50 seconds separated each repeated sprint. Forearm venous plasma inosine, hypoxanthine (Hx) and uric acid concentrations were measured at rest and during 120 minutes of recovery from exercise. Urinary inosine, Hx, xanthine, and uric acid excretion were also measured before and for 24 hours after exercise. During the first 120 minutes of recovery, plasma Hx concentrations, as well as the urinary Hx and xanthine excretion rates, were higher (P < .05) with allopurinol compared with the placebo trial. In contrast, plasma uric acid concentration and urinary uric acid excretion rates were lower (P < .05) with allopurinol. The total urinary excretion of purines (inosine + Hx + xanthine + uric acid) above basal levels was higher in the allopurinol trial compared with placebo. These results indicate that the total urinary purine excretion after intermittent sprint exercise was enhanced with allopurinol treatment. Furthermore, the composition of urinary purines was markedly affected by this drug.
Assuntos
Alopurinol/farmacologia , Purinas/urina , Adulto , Exercício Físico , Humanos , Inosina/sangue , Inosina/urina , Rim/metabolismo , Masculino , Ácido Úrico/sangue , Ácido Úrico/urinaRESUMO
The pharmacokinetic parameters of peldesine (BCX-34) were investigated after single and multiple oral doses in two groups of healthy adult volunteers. The pharmacodynamic elevation of endogenous inosine and 2'-deoxyguanosine was simultaneously monitored. The first group of 8 subjects received an intravenous dose (18 mg/m2) and five oral doses (30, 63, 108, 144, and 192 mg/m2) of drug. A second group of 12 subjects received 160 mg/m2 in four and in six divided doses orally. Serial blood samples and total urine outputs were collected during dosing and for at least 24 hours after the last dose was administered. One set of samples was analyzed using high-pressure liquid chromatography/ultraviolet (LC/UV) methods, validated for intact drug in human plasma and urine samples. Another set of samples was analyzed for the biomarkers, inosine and 2'-deoxyguanosine, using high-pressure LC with either mass spectrometry (MS) or electrochemical detection (EC) methods. The pharmacokinetic parameters of inosine and 2'-deoxyguanosine were calculated using noncompartmental methods and correlated against the pharmacokinetic parameters of BCX-34. For the single-dose study, the results exhibited linear pharmacokinetics over the dose range from 30 to 144 mg/m2. The calculated terminal half-life was 3.5 +/- 1.0 h, and the absolute bioavailability of the oral formulation was approximately 51%. Analysis of urine in the first 24 hours of collection accounted for approximately 82% of the absorbed intact drug. Evaluation of the multiple-dose pharmacokinetics indicated that steady-state blood concentrations were achieved by 24 hours when the drug was administered four or six times a day. A drug dose-related elevation of plasma 2'-deoxyguanosine was observed. This phenomenon was not seen with plasma inosine levels. However, analysis of urine samples showed an increase in inosine output with an increase in the drug dose. The calculated terminal half-life of inosine and 2'-deoxyguanosine was 15.3 +/- 1.8 h and 1.3 +/- 0.1 h, respectively.
Assuntos
Inibidores Enzimáticos/farmacocinética , Guanina/análogos & derivados , Purina-Núcleosídeo Fosforilase/antagonistas & inibidores , Administração Oral , Adulto , Área Sob a Curva , Cápsulas , Desoxiguanosina/sangue , Relação Dose-Resposta a Droga , Esquema de Medicação , Feminino , Guanina/sangue , Guanina/farmacocinética , Guanina/urina , Humanos , Inosina/sangue , Inosina/urina , Masculino , Pessoa de Meia-Idade , Soluções , Fatores de TempoAssuntos
Inosina/análogos & derivados , Purina-Núcleosídeo Fosforilase/fisiologia , Animais , Antígenos CD19 , Linfócitos B/citologia , Linfócitos B/imunologia , Biomarcadores , Linfócitos T CD4-Positivos/citologia , Linfócitos T CD8-Positivos/citologia , Nucleotídeos de Desoxiguanina/metabolismo , Desoxiguanosina/urina , Guanosina/urina , Guanosina Trifosfato/metabolismo , Inosina/urina , Camundongos , Camundongos Knockout , Purina-Núcleosídeo Fosforilase/genética , Purina-Núcleosídeo Fosforilase/imunologiaRESUMO
The influence of the number of sprint bouts on purine loss was examined in nine men (age 24.8 +/- 1.6 yr, weight 76 +/- 3.9 kg, peak O(2) consumption 3.87 +/- 0.16 l/min) who performed either one (B1), four (B4), or eight (B8) 10-s sprints on a cycle ergometer, 1 wk apart, in a randomized order. Forearm venous plasma inosine, hypoxanthine (Hx), and uric acid concentrations were measured at rest and during 120 min of recovery. Urinary inosine, Hx, and uric acid excretion were also measured before and 24 h after exercise. During the first 120 min of recovery, plasma inosine and Hx concentrations, and urinary Hx excretion rate, were progressively higher (P < 0.05) with an increasing number of sprint bouts. Plasma uric acid concentration was higher (P < 0.05) in B8 compared with B1 and B4 after 45, 60, and 120 min of recovery. Total urinary excretion of purines (inosine + Hx + uric acid) was higher (P < 0. 05) at 2 h of recovery after B8 (537 +/- 59 micromol) compared with the other trials (B1: 270 +/- 76; B4: 327 +/- 59 micromol). These results indicate that the loss of purine from the body was enhanced by increasing the number of intermittent 10-s sprint bouts.
Assuntos
Purinas/sangue , Purinas/urina , Corrida/fisiologia , Adulto , Humanos , Hipoxantina/sangue , Hipoxantina/urina , Inosina/sangue , Inosina/urina , Masculino , Concentração Osmolar , Ácido Úrico/sangue , Ácido Úrico/urinaRESUMO
The two basic aims of this study were to add to the limited literature concerning Inosine as an ergogenic aid, and to determine the effects of Inosine supplementation over a period of 5 and 10 days, at a dosage of 10,000 mg.d-1 on measures associated with aerobic and anaerobic performance. Seven trained, volunteer male subjects (body mass = 63.0 +/- 8.7 kg, VO2max = 67.9 +/- 3.3 ml.kg-1.min-1) participated in this study. The subjects completed three test sessions, each comprising three tests (5 x 6-s sprint, 30-s sprint, and 20-min time trial). Supplementation was carried out in a random, double-blind manner, and the test sessions were undertaken prior to (Baseline, B), on Day 6, and on Day 11. Blood was sampled prior to supplementation as well as on Days 6 and 11 and was analyzed for uric acid and 2,3 DPG. An analysis of the data indicated no performance benefit of supplementation and no improvement in 2,3 DPG concentration. Uric acid concentration increased significantly after both Days 6 and 11 (p < 0.03 and p < 0.004, respectively). It is concluded that Inosine has no ergogenic effects but may cause possible health problems if taken over long periods of time.
Assuntos
Suplementos Nutricionais , Exercício Físico , Inosina/administração & dosagem , 2,3-Difosfoglicerato/sangue , Análise de Variância , Ciclismo , Método Duplo-Cego , Humanos , Inosina/sangue , Inosina/urina , Masculino , Ácido Úrico/sangueAssuntos
Eletroforese Capilar/métodos , Erros Inatos do Metabolismo da Purina-Pirimidina/urina , Purinas/metabolismo , Urinálise/métodos , Alantoína/urina , Animais , Desoxiguanosina/urina , Guanosina/urina , Humanos , Inosina/análogos & derivados , Inosina/urina , Mamíferos , Purina-Núcleosídeo Fosforilase/deficiência , Erros Inatos do Metabolismo da Purina-Pirimidina/diagnóstico , Valores de Referência , Espectrofotometria UltravioletaRESUMO
The levels of the five methylated nucleosides pseudouridine (psi-Urd), 1-methyladenosine (1-MeAdo), 4 acetylcytidine (4-AcCyd), 1 methylinosine (1-Melno) and 7 methylguanosine (7-MeGuo) resulting from RNA degradation were examined in the urine of rheumatoid arthritis (RA) patients. Of these five, 1-MeAdo and psi-Urd were correlated with the active phase of the disease, while two others (4-AcCyd and 1-Melno), which require further evaluation, appeared to be linked to the prognosis of the disease. As RNA turnover is closely associated with cell proliferation, including that of lymphocytes in RA, there may be a hitherto unsuspected benefit in measuring 1-MeAdo and psi-Urd as biochemical markers of RA disease activity.
Assuntos
Artrite Reumatoide/urina , Nucleosídeos/urina , Adenosina/análogos & derivados , Adenosina/urina , Idoso , Biomarcadores , Citidina/análogos & derivados , Citidina/urina , Feminino , Seguimentos , Guanosina/análogos & derivados , Guanosina/urina , Humanos , Inosina/análogos & derivados , Inosina/urina , Masculino , Pessoa de Meia-Idade , Pseudouridina/urina , Capuzes de RNA/urina , RNA Ribossômico/metabolismo , RNA de Transferência/metabolismoRESUMO
Using HPLC methods, we measured the concentrations of nucleosides and nucleotides for a patient with no purine nucleoside phosphorylase (PNP; EC 2.4.2.1) enzymatic activity. Concentrations of inosine and guanosine were abnormally high in urine and plasma, whereas guanosine diphosphate (GDP) and guanosine triphosphate (GTP) concentrations in erythrocytes were depleted. The unusual presence of deoxyribonucleosides (deoxyinosine and deoxyguanosine) and deoxyribonucleotides (dGDP and dGTP) was also notable. Thus, HPLC represents an accurate and useful tool for the study of purine metabolic disorders.
Assuntos
Cromatografia Líquida de Alta Pressão/métodos , Purina-Núcleosídeo Fosforilase/deficiência , Erros Inatos do Metabolismo da Purina-Pirimidina/metabolismo , Nucleotídeos de Desoxiadenina/sangue , Nucleotídeos de Desoxiguanina/sangue , Eritrócitos/metabolismo , Guanosina/sangue , Guanosina/urina , Guanosina Difosfato/sangue , Guanosina Trifosfato/sangue , Humanos , Lactente , Inosina/sangue , Inosina/urina , MasculinoRESUMO
We investigated the effect of glucose infusion on adenosine triphosphate degradation in skeletal muscle of patients with glycogen storage disease type VII. Three patients and six healthy subjects exercised on a bicycle ergometer twice, once with 20% glucose infusion and once with saline infusion. The glucose infusion increased plasma glucose levels to 170 to 182 mg/dl and serum insulin levels to 30 to 50 microU/ml, while it markedly decreased plasma free fatty acid levels. The exercise-induced increases in plasma ammonia, inosine, and hypoxanthine were much larger with glucose than with saline infusion in the patients. Urinary excretion of inosine and hypoxanthine with glucose infusion was twice as high as that with saline infusion. No such differences were present between glucose and saline infusion in the healthy subjects. Glucose infusion therefore accelerates the energy crisis in working muscle of patients with glycogen storage disease type VII, probably due to a decrease in fatty acid utilization.
Assuntos
Nucleotídeos de Adenina/metabolismo , Glucose/farmacologia , Doença de Depósito de Glicogênio Tipo VII/metabolismo , Músculo Esquelético/metabolismo , Adulto , Amônia/sangue , Glicemia/metabolismo , Teste de Esforço , Ácidos Graxos não Esterificados/sangue , Feminino , Glucose/administração & dosagem , Doença de Depósito de Glicogênio Tipo VII/fisiopatologia , Humanos , Hipoxantina , Hipoxantinas/sangue , Hipoxantinas/urina , Inosina/sangue , Inosina/urina , Insulina/sangue , Masculino , Músculo Esquelético/fisiologia , Músculo Esquelético/fisiopatologia , Esforço Físico , Valores de Referência , Fatores de TempoRESUMO
Analysis of urine from cancer patients by capillary gas chromatography/mass spectrometry positively identified 14 urinary nucleosides including several modified nucleosides. Levels of the modified nucleosides 1-methyl-adenosine, 2-methylguanosine, N2,N2-dimethylguanosine and 1-methylinosine as well as the total nucleoside level were elevated in the urine when a malignant tumour was present; the levels of N2,N2-dimethylguanosine were found to correlate with the stage of the cancer.
Assuntos
Biomarcadores Tumorais , Cromatografia Gasosa-Espectrometria de Massas , Guanosina/análogos & derivados , Inosina/análogos & derivados , Neoplasias/urina , Nucleosídeos/urina , Adenosina/análogos & derivados , Adenosina/urina , Feminino , Guanosina/urina , Humanos , Inosina/urina , MasculinoRESUMO
A highly sensitive enzyme-linked immunoassay (ELISA) was developed to detect and quantify the tumor marker, 1-methylinosine (m1I), in human urine. The rabbit antisera was highly specific for m1I with negligible or no inhibition by other nucleosides excreted into urine. Using the competitive ELISA, nanogram amounts of m1I were easily measured directly in urine. The assay agreed with our previous hplc analysis of m1I in urine for identifying those individuals with chronic myelogenous leukemia. Thus, this assay should greatly facilitate the quantitation of m1I as a tumor marker.
Assuntos
Biomarcadores Tumorais/urina , Ensaio de Imunoadsorção Enzimática , Inosina/análogos & derivados , Especificidade de Anticorpos , Humanos , Soros Imunes/imunologia , Inosina/imunologia , Inosina/urina , Leucemia Mielogênica Crônica BCR-ABL Positiva/urina , MicroquímicaRESUMO
We used a reversed-phase "high-performance" liquid-chromatographic system equipped with a multichannel ultraviolet spectrometric detector and a micro-computer for analyzing urine samples from patients with disorders of purine metabolism. This system recorded a series of absorption-spectrum data from a single chromatographic run and stored them for subsequent analysis. Because the retention times and ultraviolet absorption spectra of the eluates were recorded simultaneously, identification of peaks was easy and quite accurate for simultaneous quantification of orotidine, adenine, hypoxanthine, uric acid, xanthine, allopurinol (4-hydroxypyrazolo[3,4-d]pyrimidine), oxypurinol (4,6-dihydroxypyrazolo[3,4-d]pyrimidine), inosine, and 2,8-dihydroxyadenine--compounds extremely difficult or even impossible to quantify simultaneously with a conventional single-wavelength spectrometer. We used this method to investigate purine metabolites in urines from a patient with hereditary xanthinuria, three patients with 2,8-dihydroxyadenine urolithiasis, and a gouty subject taking allopurinol.