INTRODUCTION: The analysis of urinary catecholamine metabolites is a cornerstone of neuroblastoma diagnostics. Currently, there is no consensus regarding the sampling method, and variable combinations of catecholamine metabolites are being used. We investigated if spot urine samples can be reliably used for analysis of a panel of catecholamine metabolites for the diagnosis of neuroblastoma. METHODS: Twenty-four-hour urine or spot urine samples were collected from patients with and without neuroblastoma at diagnosis. Homovanillic acid (HVA), vanillylmandelic acid (VMA), dopamine, 3-methoxytyramine, norepinephrine, normetanephrine, epinephrine and metanephrine were measured by high-performance liquid chromatography coupled with fluorescence detection (HPLC-FD) and/or ultra-performance liquid chromatography coupled with electrospray tandem mass spectrometry (UPLC-MS/MS). RESULTS: Catecholamine metabolite levels were measured in urine samples of 400 neuroblastoma patients (24-hour urine, n = 234; spot urine, n = 166) and 571 controls (all spot urine). Excretion levels of catecholamine metabolites and the diagnostic sensitivity for each metabolite were similar in 24-hour urine and spot urine samples (p > .08 and >.27 for all metabolites). The area under the receiver-operating-characteristic curve (AUC) of the panel containing all eight catecholamine metabolites was significantly higher compared to that of only HVA and VMA (AUC = 0.952 vs. 0.920, p = .02). No differences were observed in metabolite levels between the two analysis methods. CONCLUSION: Catecholamine metabolites in spot urine and 24-hour urine resulted in similar diagnostic sensitivities. The Catecholamine Working Group recommends the implementation of spot urine as standard of care. The panel of eight catecholamine metabolites has superior diagnostic accuracy over VMA and HVA.
- MeSH
- chromatografie kapalinová metody MeSH
- kyselina homovanilová moč MeSH
- kyselina vanilmandlová moč MeSH
- lidé MeSH
- metanefrin moč MeSH
- neuroblastom * diagnóza MeSH
- tandemová hmotnostní spektrometrie * metody MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Novel method for the determination of a tumor marker homovanillic acid (HVA) in human urine was developed. Combination of hollow fiber - based liquid-phase microextraction (HF-LPME) and differential pulse voltammetry (DPV) at a cathodically pre-treated boron doped diamond electrode (BDDE) was applied for these purposes. Optimum conditions were: butyl benzoate as supported liquid membrane (SLM) formed on polypropylene HF, 0.1 mol L-1 HCl as donor phase, 0.1 mol L-1 sodium phosphate buffer of pH 6 as acceptor phase, and 30 min extraction time. HF-LPME-DPV concentration dependence was linear in the range from 1.2 to 100 μmol L-1. Limits of quantification (LOQ) and detection (LOD) were 1.2 and 0.4 μmol L-1, respectively. The applicability of the developed method was verified by analysis of human urine. Standard addition method was used, found HVA concentration was 13.5 ± 1.3 μmol L-1, RSD = 9.3% (n=5).
- MeSH
- kyselina homovanilová MeSH
- lidé MeSH
- mikroextrakce kapalné fáze * MeSH
- nádorové biomarkery * MeSH
- vysokoúčinná kapalinová chromatografie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
Homovanillic (HVA), vanillylmandelic (VMA), and 5-hydroxy-3-indoleacetic acid (HIAA) are cancer biomarkers and their abnormally high urinary levels can serve as a useful tool for prediction of neuroblastoma, pheochromocytoma, and carcinoid tumors. HVA and VMA are connected with the catecholamine metabolism, while HIAA is related to the tryptophan metabolism. The aim of this review is to describe the electrochemical oxidation HVA, VMA, and HIAA and summarize published voltammetric and amperometric methods suitable for the determination of these biomarkers in various biological fluids.
RATIONALE AND OBJECTIVES: Ghrelin, an orexigenic (appetite stimulating) peptide activates binding sites in the ventral tegmental area (a structure linked with the neural reward system) allowing it to participate in reward-seeking behavior. An increasing number of studies over the past few years have demonstrated ghrelin's role in alcohol, cocaine, and nicotine abuse. However, the role of ghrelin, in opioid effects, has rarely been examined. The aim of the present study was to ascertain whether a ghrelin antagonist (JMV2959) was able to inhibit markers of morphine-induced activation of the neural reward system, namely morphine-induced increase of dopamine in the nucleus accumbens and behavioral changes in rats. METHODS: We used in vivo microdialysis to determine changes of dopamine and its metabolites in the nucleus accumbens shell in rats following morphine (MO, 5, 10 mg/kg s.c.) administration with and without ghrelin antagonist pretreatment (JMV2959, 3, 6 mg/kg i.p., 20 min before MO). Induced behavioral changes were simultaneously monitored. RESULTS: JMV2959 significantly and dose dependently reduced MO-induced dopamine release in the nucleus accumbens shell and affected concentration of by-products associated with dopamine metabolism: 3-methoxytyramine (3-MT), 3,4-dihydroxyphenylacetic acid (DOPAC), and homovanillic acid (HVA). JMV2959 pretreatment also significantly reduced MO-induced behavioral stimulation, especially stereotyped behavior. CONCLUSIONS: Ghrelin secretagogue receptors (GHS-R1A) appear to be involved in the opioid-induced changes in the mesolimbic dopaminergic system associated with the reward processing.
- MeSH
- chování zvířat účinky léků MeSH
- dopamin analogy a deriváty metabolismus farmakologie MeSH
- ghrelin metabolismus MeSH
- glycin aplikace a dávkování analogy a deriváty farmakologie MeSH
- krysa rodu rattus MeSH
- kyselina 3,4-dihydroxyfenyloctová metabolismus MeSH
- kyselina homovanilová farmakologie MeSH
- mikrodialýza MeSH
- morfin aplikace a dávkování farmakologie MeSH
- nucleus accumbens účinky léků metabolismus MeSH
- odměna MeSH
- opioidní analgetika aplikace a dávkování farmakologie MeSH
- potkani Wistar MeSH
- receptory ghrelinu antagonisté a inhibitory metabolismus MeSH
- stereotypní chování účinky léků MeSH
- triazoly aplikace a dávkování farmakologie MeSH
- vztah mezi dávkou a účinkem léčiva MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
A sensitive assay method was developed for a parallel, rapid and precise determination of dopamine and its metabolites, homovanillic acid, 3-methoxytyramine and 3,4-dihydroxyphenylacetic acid, from brain microdialysates. The method consisted of a pre-treatment step, freeze-drying (lyophilization), to concentrate dopamine and its metabolites from the microdialysates, and a detection step using liquid chromatography combined with electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS). In particular, the reaction monitoring mode was selected for its extremely high degree of selectivity and the stable-isotope-dilution assay for its high precision of quantification. The developed method was characterized by the following parameters: the precision of the developed method was determined as ≥88.6% for dopamine, ≥89.9% for homovanillic acid, ≥86.1% for 3-methoxytyramine and ≥88.1% for 3,4-dihydroxyphenylacetic acid; the mean accuracy was determined as ≥88.2% for dopamine, ≥88.3% for homovanillic acid, ≥85.9% for 3-methoxytyramine and ≥88.6% for 3,4-dihydroxyphenylacetic acid. The developed method was compared to (1) other combinations of pre-treatment methods (solid phase extraction and nitrogen stripping) with LC-MS and (2) another detection method, liquid chromatography, with electrochemical detection. The novel developed method using combination of lyophilization with LC-ESI-MS/MS was tested on real samples obtained from the nucleus accumbens of rat pups after an acute methamphetamine administration. It was proven that the developed assay could be applied to both a simultaneous analysis of all four substrates (dopamine, homovanillic acid, 3-methoxytyramine and 3,4-dihydroxyphenylacetic acid) in microdialysis samples acquired from the rat brain and the monitoring of their slight concentration changes on a picogram level over time following methamphetamine stimulus.
- MeSH
- chromatografie kapalinová metody MeSH
- dopamin analogy a deriváty analýza metabolismus MeSH
- krysa rodu rattus MeSH
- kyselina 3,4-dihydroxyfenyloctová analýza metabolismus MeSH
- kyselina homovanilová analýza metabolismus MeSH
- lineární modely MeSH
- lyofilizace MeSH
- methamfetamin aplikace a dávkování MeSH
- mikrodialýza MeSH
- nucleus accumbens chemie metabolismus MeSH
- potkani Wistar MeSH
- reprodukovatelnost výsledků MeSH
- senzitivita a specificita MeSH
- stabilita léku MeSH
- tandemová hmotnostní spektrometrie metody MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Urine contains a variety of organic and inorganic chemicals including a number of natural fluorophores. Most of them are formed by tryptophan metabolites. But there are also metabolites of riboflavin, catecholamines and porphyrins. The alternation in the autofluorescence of urine and the alternation in the concentration of these substances are developed by both physiological and pathological changes such as disorder of body metabolism, dietary intake, age and etc. In this work we present fluorescent properties of chosen urine fluorophores – i.e. 5-hydroxyindole-3-acetic acid (5-HIAA), indoxyl sulphate (urine indican), serotonin (5-HT), vanillylmandelic (VMA) and homovanillic (HVA) acids typical for various diseases. Differences of fluorescent parameters of individual fluorophores measured in vitro in the water solutions and in natural environment of urine are significant and can lead to false results and conclusions. Therefore, we present the most common influence that can occur in urine (e.g. pH, ionic strength, proteins, and other fluorophores). The aim is to elaborate the exact “know-how” for direct complex fluorescent measurement in urine related to particular diagnoses.
Methamphetamine is a commonly abused psychostimulant that causes addiction and is often abused by pregnant women. Acute or chronic administration of methamphetamine elevates the levels of the extracellular monoamine neurotransmitters, such as dopamine. The aim of the present study was to show whether prenatal exposure to methamphetamine (5mg/kg, entire gestation) or saline in Wistar rats induces changes in dopamine levels and its metabolites in the nucleus accumbens, and in behavior (locomotor activity, rearing, and immobility) after the administration of a challenge dose of methamphetamine (1mg/kg) or saline in male offspring. We found that adult offspring prenatally exposed to methamphetamine had higher basal levels of dopamine (about 288%), dihydroxyphenylacetic acid (about 67%) and homovanillic acid (about 74%) in nucleus accumbens. An increased basal level of dopamine corresponds to lower basal immobility in offspring prenatally exposed to methamphetamine. The acute injection of methamphetamine in adulthood increased the level of dopamine in the nucleus accumbens, which is related to an increase of locomotion and rearing (exploration). In addition, prenatally methamphetamine-exposed rats showed higher response to the challenge dose of methamphetamine, when compared to prenatally saline-exposed rats. In conclusion, rats exposed to methamphetamine in utero have shown changes in the mesolimbic dopaminergic system and were more sensitive to the administration of the acute dose of methamphetamine in adulthood.
- MeSH
- čas MeSH
- chování zvířat fyziologie účinky léků MeSH
- dopamin metabolismus MeSH
- inhibitory vychytávání adrenergních neurotransmiterů toxicita MeSH
- krysa rodu rattus MeSH
- kyselina 3,4-dihydroxyfenyloctová metabolismus MeSH
- kyselina homovanilová metabolismus MeSH
- methamfetamin toxicita MeSH
- modely nemocí na zvířatech MeSH
- nervové dráhy patofyziologie růst a vývoj účinky léků MeSH
- nucleus accumbens patofyziologie růst a vývoj účinky léků MeSH
- pátrací chování fyziologie účinky léků MeSH
- pohybová aktivita fyziologie účinky léků MeSH
- tegmentum mesencephali - area ventralis patofyziologie růst a vývoj účinky léků MeSH
- těhotenství MeSH
- vztah mezi dávkou a účinkem léčiva MeSH
- zpožděný efekt prenatální expozice metabolismus patofyziologie patologie MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- mužské pohlaví MeSH
- těhotenství MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- práce podpořená grantem MeSH
- MeSH
- dospělí MeSH
- hypoxanthin MeSH
- ischemie mozku * diagnóza MeSH
- kyselina homovanilová MeSH
- kyselina hydroxyindoloctová MeSH
- kyselina močová MeSH
- lidé středního věku MeSH
- lidé MeSH
- malondialdehyd MeSH
- methoxyhydroxyfenylglykol MeSH
- mozkomíšní mok * enzymologie chemie MeSH
- senioři MeSH
- taurin MeSH
- tryptofan MeSH
- vysokoúčinná kapalinová chromatografie MeSH
- xanthin MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- senioři MeSH
- MeSH
- beta-adrenergní receptory fyziologie MeSH
- dihydroalprenolol metabolismus MeSH
- glykemický clamp MeSH
- hypertriglyceridemie genetika patofyziologie MeSH
- inzulinová rezistence MeSH
- krevní glukóza metabolismus MeSH
- krysa rodu rattus MeSH
- kyselina homovanilová moč MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- mužské pohlaví MeSH
- zvířata MeSH
