Sensing of Catecholamine in Human Urine Using a Simple Colorimetric Assay Based on Direct Melanochrome and Indolequinone Formation.

We used the first enzyme-free synthesis and stabilization of soluble melanochrome (MC) and 5,6-indolequinone (IQ) derived from levodopa (LD), dopamine (DA), and norepinephrine (NE) oxidation to develop a simple colorimetric assay for catecholamine detection in human urine, also elucidating the time-dependent formation and molecular weight of MC and IQ using UV-Vis spectroscopy and mass spectrometry. The quantitative detection of LD and DA was achieved in human urine using MC as a selective colorimetric reporter to demonstrate the potential assay applicability in a matrix of interest in therapeutic drug monitoring (TDM) and in clinical chemistry. The assay showed a linear dynamic range between 5.0 mg L-1 and 50.0 mg L-1, covering the concentration range of DA and LD found in urine samples from, e.g., Parkinson's patients undergoing LD-based pharmacological therapy. The data reproducibility in the real matrix was very good within this concentration range (RSDav% 3.7% and 6.1% for DA and LD, respectively), also showing very good analytical performances with the limits of detection of 3.69 ± 0.17 mg L-1 and 2.51 ± 0.08 mg L-1 for DA and LD, respectively, thus paving the way for the effective and non-invasive monitoring of dopamine and levodopa in urine from patients during TDM in Parkinson's disease.

Pediatric total fractionated metanephrines: age-related reference intervals in spot urine.

BACKGROUND: Sparse metanephrines reference intervals in pediatric populations are available and different study designs and technologies/ assays used in these studies lead to hardly transferable data from a laboratory to another. The aim of this study was to update pediatric reference intervals of total fractionated metanephrines in spot urine samples, using a commercial extraction kit run on a specific high-pressure liquid chromatograph coupled with an electrochemical detector. METHODS: Four hundred and fifty-two spot pediatric urinary samples previously submitted to urinalysis were consecutively included in the study with the exclusion of children's samples with diagnosis or clinical suspicion of paraganglioma/pheochromocytoma, kidney diseases and arterial hypertension. Urinary metanephrine, normetanephrine and 3-methoxytyramine were extracted with ClinRep® HPLC Complete kit and run on HPLC Prominence liquid chromatograph LC-20AT (Shimadzu Italia S.r.l. Milan, Italy) coupled with Decade II electrochemical detector (Antec Scientific, Zoeterwoude, the Nederlands, provided by Alfatech S.r.l., Genoa, Italy). Results were expressed as the ratio analyte-to-creatinine. RESULTS: Any of the three analytes required a repartition by gender (metanephrine P=0.27; normetanephrine P=0.90 and 3-methoxytyramine P=0.18). A significant statistically inversely proportional relation with age was found for metanephrine (P<0.0001; ρ=-0.72), normetanephrine (P<0.0001; ρ=-0.75) and 3-methoxytyramine (P<0.0001; ρ=-0.83). Reference intervals were calculated as function of age. CONCLUSIONS: This study provides pediatric reference intervals for urinary fractionated total metanephrines in spot urine calibrated on a specific instrumentation and extraction commercial kit.

The effect of acid use as a preservative on the results of biochemical tests measured in 24-h urine.

Twenty-four-hour urine measurements play a crucial role in the diagnosis, follow-up and treatment of various diseases. There are different approaches to the collection of urine in patients who need to collect multiple urine samples at a time, especially in hospitals with heavy workloads. In this study, we compared the sodium, potassium, chloride, amylase, calcium, creatinine, phosphorus, microalbumin, protein, magnesium, urea, uric acid, adrenaline, noradrenaline, dopamine, metanephrine, normetanephrine, vanillylmandelic acid, 5-hydroxyindoleacetic acid and homovanillic acid results of 24-h urine samples analyzed immediately without acid addition, which we accepted as the reference and baseline measurement, with the results of the samples analyzed after waiting for 24 h without acid addition, analyzed immediately with acid addition and analyzed after waiting for 24 h with acid addition. Chloride, microalbumin, amylase and protein tests, which are recommended to be measured in the sample without preservatives, are affected by acid addition. Adrenaline, noradrenaline and dopamine, which are the tests recommended to be measured in acid-added urine are degraded in the samples without acid, and the levels of metanephrine and normetanephrine were not significantly degraded in the absence of preservatives.

Dual-mode detection of dopamine based on 0D/2D/2D CuInS2/ZnS quantum dot-black phosphorous nanosheet-TiO2 nanosheet nanocomposites.

In this work, we designed new dual-mode "turn-on" electrochemical (EC) and photoelectrochemical (PEC) sensors for the detection of dopamine (DA) based on 0D/2D/2D CuInS2/ZnS quantum dot (QD)-black phosphorous nanosheet (BPNS)-TiO2 nanosheet (TiO2NS) nanocomposites. QDs can not only improve the photocurrent of the developed PEC sensors, but also provide the electrochemical signal in the EC detection. BPNSs as p-type semiconductor with high conductive properties work as electron acceptors and are utilized to improve the sensitivity of the DA PEC and EC sensors. Under irradiation of visible light or the applied voltage, DA is both excited and releases electrons, realizing "turn-on" detection. The PEC sensors have a linear range of 0.1-100 µM with a lower detection limit of 0.028 µM. For the EC detection, BPNSs can accelerate electron transfer which attribute to its excellent conductivity. In the range of 1-200 µM, the working curve of DA detection by the EC sensors was established and the detection limit is 0.88 µM. Comparing the two methods, the PEC sensors have a lower detection limit, and the EC sensors have a wider monitoring range. The dual-mode sensors of EC and PEC pave an effective way for the detection in biological and medical fields.

Urinary 3-Methoxytyramine Is a Biomarker for MYC Activity in Patients With Neuroblastoma.

PURPOSE: Elevated urinary 3-methoxytyramine (3MT) level at diagnosis was recently put forward as independent risk factor for poor prognosis in neuroblastoma. Here, we investigated the biologic basis underlying the putative association between elevated 3MT levels and poor prognosis. METHODS: Urinary 3MT levels and prognosis were investigated in both retrospective Italian (N = 90) and prospective Dutch (N = 95) cohorts. From the Dutch Cancer Oncology Group cohort (N = 122), patients with available urinary 3MT and gene expression data (n = 90) were used to generate a 3MT gene signature. The 3MT gene signature score was then used to predict survival outcome in the Children's Oncology Group (N = 247) and German Pediatric Oncology Group (N = 498) cohorts and compared with other known gene signatures. Immunohistochemistry of MYCN and dopamine ß-hydroxylase proteins was performed on primary tumors. RESULTS: Elevated urinary 3MT levels were associated with poor prognosis in a retrospective cohort and a prospective cohort. Moreover, elevated urinary 3MT levels were associated with eight differentially expressed genes, providing a 3MT gene signature that successfully predicted poor clinical outcome. Even among low-risk patients, high 3MT signature score was associated with poor 5-year overall survival (72% v 99% among low-risk patients with a low 3MT signature score), and the 3MT signature score was correlated with MYC activity in the tumor (R = 82%, P < .0001). Finally, a strong MYCN and weak dopamine ß-hydroxylase staining of tumors derived from patients with elevated urinary 3MT levels was observed, linking MYC activity in the tumor to both catecholamine biosynthesis and elevated urinary 3MT levels. CONCLUSION: Elevated urinary 3MT is a promising biomarker for poor prognosis and reflects increased MYC activity in the tumor. Therefore, urinary 3MT levels should be measured at diagnosis and may assist in assessing risk.

Clinical utility of urinary levels of catecholamines and their fraction ratios related to heart rate and thyroid function.

Urinary catecholamines (CAs) have been examined for the screening of pheochromocytomas. The decision to perform screening is based on symptoms suggesting secondary hypertension or hyperactivities of the sympathetic nervous system. To elucidate the usefulness of urinary fractions and ratios of CAs, 79 patients in whom 24-h excretions of urinary CAs including adrenaline (AD), noradrenaline (NA) and dopamine (DA) had been examined from 2015 until 2020 were retrospectively analyzed. There were no significant differences in urinary CA levels between two age groups, gender groups and two BMI groups. Patients with histories of preexisting hypertension and diabetes showed significantly higher levels of urinary NA excretion, and the urinary ratio of NA/DA was also increased in the patients with a history of hypertension. Heart rate (HR) was significantly correlated with the urinary ratio of NA/DA. Serum free thyroxine (FT4) concentration and ratio of FT4/thyrotropin (TSH) were correlated with the level of urinary AD. The levels of TSH and FT4/TSH showed negative and positive correlations, respectively, with the urinary NA/DA ratio. Thus, increases of HR are related to the enhanced conversion of DA to NA and increased thyroid hormones are involved in the increase in urinary AD and the conversion of DA to NA. History of lifestyle-related diseases and changes of HR and thyroid functions need to be considered for the evaluation of urinary CAs and their ratios.

Effect of Exhaustive Training or Forced Immobilization on Physiological Condition and Main Metabolic and Stress Markers of Wistar Male Rats.

For correct and reliable experimental in vivo assessment of antistress effect of various bioactive substances, appropriate biomodels reproducing stress and organism response to stress in laboratory animals should be chosen. We chose treadmill test for simulating exhaustive physical load and forced immobilization accompanied by disorders of physiological and psychological condition. Verification of the models used indicates their wide applicability for testing certain biological manifestations under reproduced stress exposure.

Urinary Dopamine Excretion Rate Decreases during Acute Dietary Protein Deprivation and Is Associated with Increased Plasma Pancreatic Polypeptide Concentration.

Background: Dopamine, a key neurotransmitter in the autonomic nervous system participating in the homeostatic balance between sympathetic and parasympathetic divisions, is involved in food intake regulation. Objective: We investigated whether dopamine is altered by acute fasting or overfeeding diets with varying macronutrient content. Design: Ninety-nine healthy subjects underwent 24-h dietary interventions including eucaloric feeding, fasting, and five different overfeeding diets in a crossover design. Overfeeding diets (200% of eucaloric requirements) included one diet with 3%-protein (low-protein high-fat overfeeding-LPF: 46%-fat), three diets with 20%-protein, and a diet with 30%-protein (44%-fat). Urine was collected for 24 h and urinary dopamine concentration was quantified by high-performance liquid chromatography. Plasma pancreatic polypeptide (PP) concentration, an indirect marker of parasympathetic activity, was measured prior to and after each diet after an overnight fast. Results: During 24-h of fasting, dopamine decreased on average by ~14% compared to eucaloric conditions, whereas PP increased by two-fold (both p < 0.001). Lower dopamine during 24-h fasting correlated with increased PP (r = -0.40, p < 0.001). Similarly, on average urinary dopamine decreased during LPF by 14% (p < 0.001) and lower dopamine correlated with increased PP (r = -0.31, p = 0.01). No changes in dopamine and PP concentrations were observed during other overfeeding diets (all p > 0.05). Conclusions: Dopamine concentrations decrease during short-term fasting and overfeeding with a low-protein diet. As both dietary conditions have in common protein deficit, the correlation between dopamine and PP suggests a compensatory mechanism underlying the shift from sympathetic to parasympathetic drive during dietary protein deprivation.

A covalent organic polymer-TiO2/Ti3C2 heterostructure as nonenzymatic biosensor for voltammetric detection of dopamine and uric acid.

Heterostructures have potential to blend the advantages of each material, even exhibiting the evolutionary performance due to synergistic effects. Herein, covalent organic polymers (NUF) are integrated with a TiO2/Ti3C2Tx nanocomposite (TiO2/TiCT) to form TiO2/TiCT-NUF heterojunctions as an enlarged nonenzymatic biosensor for dopamine (DA) and uric acid (UA). Detection is performed by differential pulse voltammetry (DPV). The TiO2/TiCT/NUF exhibits high sensing activity with low detection limits of 0.2 and 0.18 nM (S/N = 3) in the concentration ranges from 0.002 to 100 µM and 0.001 to 60 µM for simultaneous determination of DA and UA, respectively. In addition, the TiO2/TiCT/NUF provides good selectivity and reproducibility for DA and UA detection in urine and serum samples with recoveries of 98.4 to 100.9%. The proposed heterojunctions manifest an intriguing potential as a candidate of an electrochemical sensor for sole and simultaneous detection of DA and UA.

Smoking Accelerates Atrioventricular Conduction in Humans Concordant with Increased Dopamine Release.

Smoking is associated with cardiac arrhythmia, stroke, heart failure, and sudden cardiac arrest, all of which may derive from increased sympathetic influence on cardiac conduction system and altered ventricular repolarization. However, knowledge of the effects of smoking on supraventricular conduction, and the role of the sympathetic nervous system in them, remains incomplete. Participants with intermediate-high cardiovascular disease risk were measured for urinary catecholamines and cotinine, and 12-lead electrocardiograms (ECGs) were measured for atrial and atrioventricular conduction times, including P duration, PR interval, and PR segment (lead II), which were analyzed for associations with cotinine by generalized linear models. Statistical mediation analyses were then used to test whether any significant associations between cotinine and atrioventricular conduction were mediated by catecholamines. ECG endpoints and urinary metabolites were included from a total of 136 participants in sinus rhythm. Atrial and atrioventricular conduction did not significantly differ between smokers (n = 53) and non-smokers (n = 83). Unadjusted and model-adjusted linear regressions revealed cotinine significantly and inversely associated with PR interval and PR segment, but not P duration. Dopamine, norepinephrine, and epinephrine all inversely associated with PR interval, whereas only dopamine was also inversely associated with PR segment (p < 0.05). Dopamine and norepinephrine (but not epinephrine) also associated positively with cotinine. Dopamine mediated the relationship between cotinine and PR interval, as well as the relationship between cotinine and PR segment. Smoking is associated with accelerated atrioventricular conduction and elevated urinary dopamine and norepinephrine. Smoking may accelerate atrioventricular nodal conduction via increased dopamine production.

Early pregnancy metabolites predict gestational diabetes mellitus: implications for fetal programming.

BACKGROUND: Aberrant fetal programming in gestational diabetes mellitus seems to increase the risk of obesity, type 2 diabetes, and cardiovascular disease. The inability to accurately identify gestational diabetes mellitus in the first trimester of pregnancy has thwarted ascertaining whether early therapeutic interventions reduce the predisposition to these prevalent medical disorders. OBJECTIVE: A metabolomics study was conducted to determine whether advanced analytical methods could identify accurate predictors of gestational diabetes mellitus in early pregnancy. STUDY DESIGN: This nested observational case-control study was composed of 92 gravidas (46 in the gestational diabetes mellitus group and 46 in the control group) in early pregnancy, who were matched by maternal age, body mass index, and gestational age at urine collection. Gestational diabetes mellitus was diagnosed according to community standards. A comprehensive metabolomics platform measured 626 endogenous metabolites in randomly collected urine. Consensus multivariate criteria or the most important by 1 method identified low-molecular weight metabolites independently associated with gestational diabetes mellitus, and a classification tree selected a subset most predictive of gestational diabetes mellitus. RESULTS: Urine for both groups was collected at a mean gestational age of 12 weeks (range, 6-19 weeks' gestation). Consensus multivariate analysis identified 11 metabolites independently linked to gestational diabetes mellitus. Classification tree analysis selected a 7-metabolite subset that predicted gestational diabetes mellitus with an accuracy of 96.7%, independent of maternal age, body mass index, and time of urine collection. CONCLUSION: Validation of this high-accuracy model by a larger study is now needed to support future studies to determine whether therapeutic interventions in the first trimester of pregnancy for gestational diabetes mellitus reduce short- and long-term morbidity.

FeMn layered double hydroxides: an efficient bifunctional electrocatalyst for real-time tracking of cysteine in whole blood and dopamine in biological samples.

A peculiar clock-regulated design of FeMn-LDHs (FMH) with specific physiochemical attributes has been developed and used for highly sensitive detection of cysteine (CySH) and dopamine (DA). The FMH nanoparticles were synthesized via a facile hydrothermal approach clocked at various (6 h, 12 h and 18 h) operating periods. Under optimal conditions, FMH were obtained in three unique morphologies such as hexagonal plate like, cubic, and spherical structures corresponding to the clocked periods of 6 h, 12 h, and 18 h, respectively. Among these, FMH-12 h possess the minimal particle size (54.45 nm), a large surface area (7.60 m2 g-1) and the highest pore diameter (d = 4.614 nm). In addition to these superior physiochemical attributes, the FMH nanocubes exhibit excellent electrochemical behaviors with the lowest charge transfer resistance (Rct; 96 Ω), a high heterogeneous rate constant (7.81 × 10-6 cm s-1) and a good electroactive surface area (0.3613 cm2), among the three. The electrochemical biosensor based on the FMH nanocubes exhibits a remarkable catalytic activity toward CySH and DA with a low detection limit (9.6 nM and 5.3 nM) and a broad linear range (30 nM-6.67 mM and 20 nM-700 µM). The FMH based biosensor is also feasible for the real-world detection of CySH in whole blood and DA in biological fluids with satisfactory results. The proposed sensor possessed high selectivity, good repeatability, and reproducibility toward CySH and DA sensing.

Detection of spot urinary free metanephrines and 3-methoxytyramine with internal reference correction for the diagnosis of pheochromocytomas and paragangliomas.

Detection of normetanephrine (NMN), metanephrine (MN) and 3-methoxytyramine (3-MT) could be used to diagnose pheochromocytomas and paragangliomas (PPGLs). The accuracy for the diagnosis of PPGLs is only 6% by virtue of the classic symptom triad. In addition, false-positive results were found using plasma free MNs as biomarkers. Spot urinary free metanephrines (MNs) presented high specificity for PPGLs diagnosis in our previous work by HPLC with the electrochemical detection. Whereas, MNs and creatinine (Cr) need to be detected separately. A simple and specific method was urgently needed for the diagnosis of PPGLs. Here, we established a new HPLC method for spot urinary free MNs and 3-MT by the fluorescence detection and Cr by the ultraviolet detection simultaneously. It was worth mentioning that Cr for the virtue of being fairly constant in a given subject was used as an internal reference correction to eliminate the effect of spot urine volume for the diagnosis of PPGLs. Thirty-seven patients with PPGLs and 164 control subjects were detected by the established method and the peak area ratios of MNs and 3-MT to Cr were used innovatively for the diagnosis of PPGLs. The results showed acceptable precisions and recoveries. The sensitivities of the method were 94.6%, 91.9% and 86.5% and the specificities were 96.3%, 93.9% and 82.3%, respectively by the peak area of NMN/Cr, MN/Cr and 3-MT/Cr for the diagnosis. The established method provides a promising way for simple, rapid and accurate diagnosis of PPGLs.

Photo-electrochemical detection of dopamine in human urine and calf serum based on MIL-101 (Cr)/carbon black.

A new photo-electrochemical sensor based on MIL-101(Cr) MOF/carbon black (CB) is fabricated and characterized. By using differential pulse voltammetry, dopamine (DA) can be effectively detected using a photo-electrochemical MIL-101(Cr)/CB sensor under visible light. The CB acts as the electron bridge to combine with the large specific surface area and photo-catalytic feature of MOF, which contribute to the improvements of sensitivity of DA detection. The concentration of the catalyst, pH value, accumulation potential, and accumulation time were also optimized. Furthermore, the electrochemical performances of MIL-101(Cr)/CB sensor was investigated by cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS), scan rate, electrochemically active surface area (ECSA), and amperometric responses. A detection limit of 0.38 nM (LOD = 3 sb/S, sb = 0.028) and a working range of 1 nM to 2.22 µM has been achieved. The MIL-101(Cr)/CB sensor exhibits excellent reproducibility, stability, and selectivity and also has satisfactory recovery rate for the analysis of real samples including calf serum and human urine. Graphical abstract.

Simultaneous and sensitive determination of ascorbic acid, dopamine and uric acid via an electrochemical sensor based on PVP-graphene composite.

A method with high sensitivity, good accuracy and fast response is of ever increasing importance for the simultaneous detection of AA, DA and UA. In this paper, a simple and sensitive electrochemical sensor, which based on the polyvinylpyrrolidone (PVP)-graphene composite film modified glassy carbon electrode (PVP-GR/GCE), was presented for detecting ascorbic acid (AA), dopamine (DA) and uric acid (UA) simultaneously. The PVP-GR/GCE has excellent electrocatalytic activity for the oxidation of AA, DA and UA. The second-order derivative linear sweep voltammetry was used for the electrochemical measurements. The peak potential differences of DA-AA, DA-UA, and UA-AA (measured on the PVP-GR/GCE) were 212, 130 and 342 mV respectively. Besides, the over potential of AA, DA and UA reduced obviously, so did the peak current increase. Under the optimum conditions, the linear ranges of AA, DA and UA were 4.0 µM-1.0 mM, 0.02-100 µM, and 0.04-100 µM, respectively. The detection limits were 0.8 µM, 0.002 µM and 0.02 µM for AA, DA, and UA. The electrochemical sensor presented the advantages of high sensitivity and selectivity, excellent reproducibility and long-term stability. Furthermore, the sensor was successfully applied to the analysis of real samples.

Electrochemical sensor based on CuSe for determination of dopamine.

A simple binary copper selenide, CuSe nanostructure, has been investigated as electrochemical sensor for dopamine detection. The hydrothermally synthesized and electrodeposited CuSe nanostructures showed high sensitivity for dopamine detection with low limit of detection (LOD). A sensitivity of 26 µA/µM.cm2 was obtained with this electrochemical sensor which is ideal to detect even small fluctuations in the transient dopamine concentration. Apart from high sensitivity and low LOD, the dopamine oxidation on the catalyst surface also occurred at a low applied potential (< 0.18 V vs Ag|AgCl), thereby significantly increasing selectivity of the process specifically with respect to ascorbic and uric acids, which are considered to be the most prominent interferents for dopamine detection. Electrochemical redox tunability of the catalytic Cu center along with low coordination geometry is believed to enhance the rate of dopamine attachment and oxidation on the catalyst surface thereby reducing the applied potential. The presence of Cu also increases conductivity of the catalyst composite which further improves the charge transfer thus increasing the sensitivity of the device. This is the first report of electrochemical dopamine sensing with a simple binary selenide comprising earth-abundant elements and can have large significance in designing efficient sensors that can be transformative for understanding neurodegenerative diseases further. Graphical abstract.

Dopamine-induced photoluminescence quenching of bovine serum albumin-capped manganese-doped zinc sulphide quantum dots.

The direct detection of dopamine (DA) in human body fluids is a great challenge for medical diagnostics of neurological disorders like Parkinson's disease, Alzheimer's disease, senile dementia, and schizophrenia. In this work, a simple and turn off luminescence sensing of DA based on bovine serum albumin (BSA)-capped manganese-doped zinc sulphide quantum dots (Mn:ZnS/BSA QDs) is developed. The Mn:ZnS/BSA QDs were synthesized by a chemical co-precipitation method. Due to the special interaction of DA with BSA and metal ions, Mn:ZnS/BSA QDs can serve as an effective sensing platform for DA. The luminescence of Mn:ZnS/BSA QDs decreased linearly with increasing concentration of DA in the range from 6.6 to 50.6 nM. The limit of detection is 2.02 nM. The driving force for the luminescence quenching is partly provided by ground-state complex formation of QDs with DA. The photo-induced electron transfer from the conduction band of QDs to oxidized dopamine (quinone) also favors quenching. The Mn:ZnS/BSA QDs are barely interfered with by other competing biomolecules except catecholamine neurotransmitter like epinephrine. Moreover, this method is used in the analysis of DA-spiked human serum and human urine samples and good recovery percentages are found. To assess the utility of the developed sensor, paper strip assay was also successfully conducted. Graphical abstract.

Label-free liquid crystal-based biosensor for detection of dopamine using DNA aptamer as a recognition probe.

We present a label-free liquid crystal-based biosensor for the detection of dopamine (DA) in aqueous solutions using dopamine-binding aptamers (DBA) as recognition elements. In this system, the dimethyloctadecyl [3-(trimethoxysilyl) propyl] ammonium chloride (DMOAP) self-assembled monolayers immobilized on glass slides support the long alkyl chains that keep the liquid crystal (LC) molecules in a homeotropic orientation. Glutaraldehyde (GA) is used as a cross-linker to immobilize DBA onto the surface of glass slides. The specific binding of DA and DBA disrupts the homeotropic orientation of LCs, thereby inducing a change in the orientation from homeotropic to a random alignment. This orientation change can be converted and visualized simply as a transition from a dark optical LC image to a brighter image under a polarized optical microscope (POM), enabling the detection of DA. The developed LC-based aptasensor shows a good linear optical response towards DA in the very wide range of 1 pM-10 µM (0.19 pg/mL to 1.9 µg/mL) and has a very low detection limit of 10 pM (∼1.9 pg/mL). The biosensor also exhibited satisfactory selectivity and could be successfully applied to detect DA in human urine. The proposed LC-based aptamer sensing method offers a simple, rapid, highly sensitive and selective, and a label-free method for the analysis of DA in real clinical samples.

Random 'spot' urinary metanephrines compared with 24-h-urinary and plasma results in phaeochromocytomas and paragangliomas.

BACKGROUND: In patients with phaeochromocytomas or paragangliomas (PPGLs), 24-h urine collections for metanephrines (uMNs) are cumbersome. OBJECTIVE: To evaluate the diagnostic utility of ratios to creatinine of 'spot' uMNs. METHODS: Concentrations of uMNs and plasma metanephrines (pMNs) were measured by HPLC-mass-spectrometry. We retrospectively compared correlations of 24-h-urine output and ratio to creatinine in historical specimens and prospectively assessed 24-h and contemporaneous spot urines and, where possible, pMNs. Using trimmed log-transformed values, we derived reference intervals based on age and sex for spot urines. We used multiples of upper limit of normal (ULNs) to compare areas under curves (AUCs) for receiver-operator characteristic curves of individual, and sum and product of, components. RESULTS: In 3143 24-h-urine specimens on 2416 patients, the correlation coefficients between the ratios and outputs of metanephrine, normetanephrine and 3-methoxytyramine in 24-h urines were 0.983, 0.905 and 0.875, respectively. In 96 patients, the correlations between plasma concentrations, urine output and ratios in spot specimens were similar to those for raw output or ratios in 24-h specimens. Of the 160 patients with PPGLs, the CIs for AUCs for individual metabolites overlapped for all four types of measurement, as did those for the sum of the multiple ULNs although these were slightly higher (AUC for spot urine: 0.838 (0.529-1), plasma: 0.929 (0.874-0.984) and output: 0.858 (0.764-0.952)). CONCLUSIONS: Ratios of fractionated metanephrines to creatinine in spot urine samples appear to have a similar diagnostic power to other measurements. The ease of spot urine collection may facilitate diagnosis and follow-up of PPGLs through improved patient compliance.

Blood and urine biomarkers associated with long-term respiratory dysfunction following neonatal hyperoxia exposure: Implications for prematurity and risk of SIDS.

Former preterm infants, many of whom required supplemental O2 support, exhibit sleep disordered breathing and attenuated ventilatory responses to acute hypoxia (HVR) beyond their NICU stay. There is an increasing awareness that early detection of biomarkers in biological fluids may be useful predictors/identifiers of short- and long-term morbidities. In the present study, we identified serotonin (5-HT), dopamine (DA) and hyaluronan (HA) as three potential biomarkers that may be increased by neonatal hyperoxia and tested whether they would be associated with an impaired HVR in a rat model of supplemental O2 exposure. Neonatal rats (postnatal age (P) 6 days, P6) exposed to hyperoxia (40% FIO2, 24 h/day between P1-P5 days of age) exhibited an attenuated early (1 min), but not the late (4-5 min) phase of the HVR compared to normoxia control rats; the attenuated early phase HVR was associated with increased levels of DA (urine and serum), 5-HT (platelet poor plasma only, PPP), and HA (serum only). At P21, both the early and late phases of the HVR were attenuated, but serum and urine levels of all 3 biomarkers were similar to age-matched control rats. These data indicate that changes in several serum and/or urine biomarkers (5-HT, DA, and HA) following short-term (days) neonatal hyperoxia can signify long-term (weeks) respiratory control dysfunction. Further studies are needed to determine whether early detection of similar biomarkers could be convenient predictors of increased risk of abnormalities in respiratory control including sleep disordered breathing in former preterm infants who had received prior supplemental O2 and who might also be at increased risk of SIDS.