An electrochemical microbiosensor for serotonin based on surface imprinted layer coordinated bimetal functionalized acupuncture needle.

Serotonin (5-hydroxytryptamine, 5-HT) is a pivotal monoamine neurotransmitter, which is widely distributed in human brain for biological, physical and psychopathological processes. The content of 5-HT can support diagnose of various diseases. To selectively detect 5-HT is very important in clinical medicine. Here, a novel microbiosensor for 5-HT is studied on acupuncture needle. Molecularly imprinted film enwrapped 5-HT was electropolymerized onto bimetallic gold/platinum (Au/Pt) nanoparticles on acupuncture needle microelectrode (ANME). Au/Pt nanostructure exhibited active sites to catalyze the oxidation of 5-HT and bind the generated polymer. 5-HT can be enwrapped by the functional monomer of pyrrole (Py) in the process of electropolymerization with suitably electroactive conformation. Comparing with interfaces of single metal or molecularly imprinted layer, synergistic microbiosensor exhibit better performance for 5-HT. 5-HT can be adsorbed and catalytically oxidized by the imprinted cavities. Under optimized conditions, the peak current linearly increases with the concentration of 5-HT from 0.03 to 500 µM, and a detection limit of 0.0106 µM is obtained. The performance of this microbiosensor is competitive with previous studies. Furthermore, the prepared microbiosensor showed effective application to analyze 5-HT in human serum and urine. Interestingly, the microbiosensor expressed the real-time monitoring ability to 5-HT from stimulated PC12 cells by K+. The microbiosensor also exhibited high selectivity, stability and reproducibility, which is promising in view of the low price, fast response and simple operation.

A label-free fluorescence aptamer sensor for point-of-care serotonin detection.

Serotonin, a pivotal neurotransmitter regulating various physiological functions, plays a crucial role in disease diagnosis, necessitating precise monitoring of its levels in biological fluids for accurate assessment. Aptamers, known for their high specificity and affinity, have emerged as innovative molecular probes for serotonin analysis. However, existing serotonin aptamer sensing platforms exhibit limitations in terms of portability and rapid detection capabilities. In this study, we introduce a novel, portable, label-free serotonin aptamer sensor utilizing a dye replacement strategy, achieving a short sample-to-result turnaround time and convenient signal readout through a smartphone. The performance of this aptamer sensor was thoroughly assessed across diverse physiological media, demonstrating robust stability in buffer, urine, and serum. Importantly, the detection limit was in the nanomolar range, emphasizing its suitability for the rapid, sensitive, and user-friendly detection of serotonin. This research pioneers an approach for the development of a point-of-care testing (POCT) system for serotonin with practical implications, particularly in resource-limited settings.

5-Hydroxytryptophan toxicity successfully treated by haemodialysis in a dog.

OBJECTIVE: To describe a case of 5-hydroxytryptophan (5-HTP) toxicity successfully treated with haemodialysis in a dog. CASE SUMMARY: A 3-year-old, male neutered Labrador Retriever, weighing 28.2 kg, presented to the emergency department approximately 4-5 h after ingesting a human supplement containing 200 mg of 5-HTP. The amount of 5-HTP ingested was estimated between 980 and 1988 mg (35-71 mg/kg). At presentation, the dog demonstrated progressive neurologic abnormalities consistent with serotonin syndrome, including altered mentation and ataxia. Due to the magnitude of the ingested dose and progression of clinical signs, extracorporeal blood purification with intermittent haemodialysis was chosen to expedite clearance of 5-HTP. High-efficiency haemodialysis was initiated, and the dog showed continued clinical improvement throughout the 5-h treatment. Clinical signs resolved completely within 12 h. Sequential blood and urine samples were obtained to document levels of both 5-HTP and serotonin. The dog was discharged 24 h after presentation with complete resolution of clinical signs. NEW OR UNIQUE INFORMATION: This is the first report documenting the serial changes in 5-HTP concentrations during treatment with haemodialysis.

Ultrasensitive determination of serotonin in human urine by a two dimensional capillary isotachophoresis-capillary zone electrophoresis hyphenated with tandem mass spectrometry.

A two-dimensional capillary isotachophoresis-capillary zone electrophoresis method hyphenated with tandem mass spectrometry was developed and validated for ultrasensitive quantification of serotonin in real human urine samples. Under optimal conditions, the separation was achieved within 12 min (including on-line sample preparation) with the limit of detection of 34 pg mL-1 (due to a large volume sample injection, here 10 µL, and isotachophoretic preconcentration). This concentration limit represents the lowest value for serotonin in comparison to other previously published separation methods employing mass spectrometry detection and applied to urine matrices. Thanks to high orthogonality, on-line concentration and clean-up effects of this approach, other excellent validation parameters such as linearity (coefficient of determination > 0.99), inter-day and intra-day precision (relative standard deviations 3.5-12.2%), accuracy (relative errors within 99-109.4%), and recovery (96-102%) could be easily obtained too. To demonstrate applicability of the method, we monitored serotonin levels in various real samples (from a healthy volunteer and clinical ones). The determined levels, normalized on the creatinine concentrations, were in the range of 6.81-12.86 ng mmol-1 creatinine This advanced method is suggested for an effective, reliable, high sample throughput, and low cost routine clinical screening or targeted metabolomic studies of serotonin in urine samples.

Evaluation and review of ways to differentiate sources of ethanol in postmortem blood.

Accurate determination of a person's blood alcohol concentration (BAC) is an important task in forensic toxicology laboratories because of the existence of statutory limits for driving a motor vehicle and workplace alcohol testing regulations. However, making a correct interpretation of the BAC determined in postmortem (PM) specimens is complicated, owing to the possibility that ethanol was produced in the body after death by the action of various micro-organisms (e.g., Candida species) and fermentation processes. This article reviews various ways to establish the source of ethanol in PM blood, including collection and analysis of alternative specimens (e.g., bile, vitreous humor (VH), and bladder urine), the identification of non-oxidative metabolites of ethanol, ethyl glucuronide (EtG) and ethyl sulfate (EtS), the urinary metabolites of serotonin (5-HTOL/5-HIAA), and identification of n-propanol and n-butanol in blood, which are known putrefaction products. Practical utility of the various biomarkers including specificity and stability is discussed.

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.

Host and Microbial Tryptophan Metabolic Profiling in Multiple Sclerosis.

Multiple sclerosis (MS) is an autoimmune disease of the central nervous system (CNS) that is associated with demyelination and neuronal loss. Over recent years, the immunological and neuronal effects of tryptophan (Trp) metabolites have been largely investigated, leading to the hypothesis that these compounds and the related enzymes are possibly involved in the pathophysiology of MS. Specifically, the kynurenine pathway of Trp metabolism is responsible for the synthesis of intermediate products with potential immunological and neuronal effects. More recently, Trp metabolites, originating also from the host microbiome, have been identified in MS, and it has been shown that they are differently regulated in MS patients. Here, we sought to discuss whether, in MS patients, a specific urinary signature of host/microbiome Trp metabolism can be potentially identified so as to select novel biomarkers and guide toward the identification of specific metabolic pathways as drug targets in MS.

Enzymatic Platforms for Sensitive Neurotransmitter Detection.

A convenient electrochemical sensing pathway was investigated for neurotransmitter detection based on newly synthesized silole derivatives and laccase/horseradish-peroxidase-modified platinum (Pt)/gold (Au) electrodes. The miniature neurotransmitter's biosensors were designed and constructed via the immobilization of laccase in an electroactive layer of the Pt electrode coated with poly(2,6-bis(3,4-ethylenedioxythiophene)-4-methyl-4-octyl-dithienosilole) and laccase for serotonin (5-HT) detection, and a Au electrode modified with the electroconducting polymer poly(2,6-bis(selenophen-2-yl)-4-methyl-4-octyl-dithienosilole), along with horseradish peroxidase (HRP), for dopamine (DA) monitoring. These sensing arrangements utilized the catalytic oxidation of neurotransmitters to reactive quinone derivatives (the oxidation process was provided in the enzymes' presence). Under the optimized conditions, the analytical performance demonstrated a convenient degree of sensitivity: 0.0369 and 0.0256 µA mM-1 cm-2, selectivity in a broad linear range (0.1-200) × 10-6 M) with detection limits of ≈48 and ≈73 nM (for the serotonin and dopamine biosensors, respectively). Moreover, the method was successfully applied for neurotransmitter determination in the presence of interfering compounds (ascorbic acid, L-cysteine, and uric acid).

Characterization of serotonin following exposure to antibiotics in white-tailed deer.

Developing baseline concentrations of serotonin in healthy white-tailed deer will allow for the development of a biomarker using non-invasive sample tissues in sick animals, for example, non-clinical cases of chronic wasting disease. It will also allow some further insight into whether the use of antibiotics as growth promoters (AGP), such as chlortetracycline, is affecting serotonin concentrations in white-tailed deer. Florfenicol and tulathromycin impacts on serotonin concentration changes were also investigated. An analytical method for the detection and confirmation of serotonin, 5-hydroxytryptamine (5-HT), in white-tailed deer tissues was developed and validated. Serum and urine samples were extracted with acetonitrile. Liquid chromatography separation was attained on a Phenomenex C18 column with a Security Guard ULTRA guard column with gradient elution using a mobile phase of 0.1% formic acid in water and 0.1% formic acid in acetonitrile. This methodology was applied to baseline (control), chlortetracycline (CTC) treated, florfenicol treated and tulathromycin treated white-tailed deer serum and urine samples.

Integration of three-phase microelectroextraction sample preparation into capillary electrophoresis.

A major strength of capillary electrophoresis (CE) is its ability to inject small sample volumes. However, there is a great mismatch between injection volume (typically <100 nL) and sample volumes (typically 20-1500 µL). Electromigration-based sample preparation methods are based on similar principles as CE. The combination of these methods with capillary electrophoresis could tackle obstacles in the analysis of dilute samples. This study demonstrates coupling of three-phase microelectroextraction (3PEE) to CE for sample preparation and preconcentration of large volume samples while requiring minimal adaptation of CE equipment. In this set-up, electroextraction takes place from an aqueous phase, through an organic filter phase, into an aqueous droplet that is hanging at the capillary inlet. The first visual proof-of-concept for this set-up showed successful extraction using the cationic dye crystal violet (CV). The potential of 3PEE for bioanalysis was demonstrated by successful extraction of the biogenic amines serotonin (5-HT), tyrosine (Tyr) and tryptophan (Trp). Under optimized conditions limits of detection (LOD) were 15 nM and 33 nM for 5-HT and Tyr respectively (with Trp as an internal standard). These LODs are comparable to other similar preconcentration methods that have been reported in conjunction with CE. Good linearity (R2 > 0.9967) was observed for both model analytes. RSDs for peak areas in technical replicates, interday and intraday variability were all satisfactory, i.e., below 14%. 5-HT, Tyr and Trp spiked to human urine were successfully extracted and separated. These results underline the great potential of 3PEE as an integrated enrichment technique from biological samples and subsequent sensitive metabolomics analysis.

Novel electrochemical biosensor for serotonin detection based on gold nanorattles decorated reduced graphene oxide in biological fluids and in vitro model.

Abnormal level of serotonin (ST) in body fluids is related to various clinical conditions including behavioral and psychotic disorders; hence its fast detection in clinically relevant ranges have tremendous importance in medical science. In view of this, we have developed a novel biosensor for ST detection using Au-nanorattles (AuNRTs)- reduced graphene oxide (rGO) nanocomposite coated on to the gold nanoparticles (AuNPs) deposited glassy carbon electrode (GCE). The nanocomposite/sensor probe was characterized using UV-Vis, TEM, SAED, EDX, AFM, and electrochemical techniques including LSV and EIS. Thereafter, the suitability of fabricated GCE/AuNPs/AuNRTs-rGO-Naf sensor probe was applied for ST determination which showed a linear dynamic range (LDR) of 3 × 10-6 - 1 × 10-3 M and the detection limit (DL) of 3.87 (±0.02) ×10-7 (RSD < 4.2%) M, which falls in the ranges of normal as well as various abnormal pathophysiological conditions. The designed sensor is successfully applied to detect ST in various real matrices viz. urine, blood serum, and in vitro model to show its direct clinical/practical applicability. Interferences due to the coexisting molecules were assessed and the long-term stability of the designed sensor was also examined which was found to be 8 weeks.

Daily fluctuation of urine serotonin and cortisol in healthy shelter dogs and influence of intraspecific social exposure.

The aim of this study was to evaluate the effects of intraspecific social exposure (socialization vs rest) and habituation factors on the levels of urinary serotonin and cortisol [corrected respectively for creatinine to give the serotonin-creatinine ratio (5-HT/Cr) and cortisol-creatinine ratio (C/Cr)] and how they fluctuate in dogs. The frequency of marking during social sessions was recorded to evaluate its relationship with physiological parameters. The effects of covariates on 5-HT/Cr and C/Cr were assessed using a linear mixed models. 5-HT/Cr values were higher at dawn than at dusk during resting days, however, this difference is less evident after socialization sessions. During rest days, there was a trend for a decrease in C/Cr between dawn and dusk, while during social exposure days there was an opposite trend. Significant interactions were found between social exposure vs rest (P = 0.0005) and social exposure vs sessions (P = 0.002). Urine marking was more frequent in male than female dogs. The frequency of urine marking also appeared to be positively associated with C/Cr. Non-invasive monitoring of physiological markers could be a useful tool in assessing behaviour modulation following intraspecific socialization exposure. C/Cr was characterized by high individual variability and interactions with examined factors. The identification of new markers such as serotonin for assessing welfare in dogs is highly desirable.

High-fat simple carbohydrate (HFSC) diet impairs hypothalamic and corpus striatal serotonergic metabolic pathway in metabolic syndrome (MetS) induced C57BL/6J mice.

OBJECTIVES: To study the effect of specially formulated high-fat simple carbohydrate diet (HFSC) on the serotonin metabolic pathway in male C57BL/6J mice. METHODS: Previous studies from our laboratory have shown that specially formulated HFSC induces metabolic syndrome in C57BL/6J mice. In the present investigation, 5-hydroxytryptophan, serotonin and 5-hydroxyindoleacetic acid were analyzed in two brain regions (hypothalamus, corpus striatum), urine and plasma of HFSC-fed mice on a monthly basis up to 5 months using high-performance liquid chromatography fitted with electrochemical detector. The data were analyzed using Graph pad Prism v7.3 by two-way ANOVA and post hoc Tukey's test (to assess the effect of time on the serotonergic metabolic pathway). RESULTS: HFSC feed was observed to lower the hypothalamic serotonergic tone as compared to the age-matched control-fed C57BL/6J mice. Although the hypothalamic serotonergic tone was unaltered over time due to consumption of diet per se, hypothalamic 5-HTP levels were observed to be lower on consumption of HFSC feed over duration of 5 months as compared to 1st month of consumption of HFSC feed. The striatal 5-HTP levels were lowered in the HFSC-fed mice after 4 months of feeding as compared to the age-matched control-fed mice. The striatal 5-HTP levels were also lower in both control and HFSC-fed mice due to consumption of the respective diet over a duration of 5 months. Increased plasma 5-HTP levels were observed due to consumption of HFSC feed over duration of 5 months in the HFSC-fed group. However, higher breakdown of serotonin was observed in both the plasma and urine of HFSC-fed C57BL/6J mice as per the turnover studies. DISCUSSION: The central and peripheral serotonergic pathway is affected differentially by both the type of diet consumed and the duration for which the diet is consumed. The hypothalamic, striatal and plasma serotonergic pathway were altered both by the type of feed consumed and the duration of feeding. The urine serotonergic pathway was affected by mainly the duration for which a particular diet was consumed. These findings may have implications in the feeding behavior, cognitive decline and depression associated with metabolic syndrome patients.

Plasma levels of glial cell marker S100B in children with autism.

Autism spectrum disorder (ASD) is a neurodevelopmental condition with increasing incidence. Recent evidences suggest glial cells involvement in autism pathophysiology. S100B is a calcium binding protein, mainly found in astrocytes and therefore used as a marker of their activity. In our study, children with autism had higher plasma concentrations of S100B compared to non-autistic controls. No association of S100B plasma levels with behavioral symptoms (ADI-R and ADOS-2 scales) was found. Plasma S100B concentration significantly correlated with urine serotonin, suggesting their interconnection. Correlation of plasma S100B levels with stool calprotectin concentrations was found, suggesting not only brain astrocytes, but also enteric glial cells may take part in autism pathogenesis. Based on our findings, S100B seems to have a potential to be used as a biomarker of human neurodevelopmental disorders, but more investigations are needed to clarify its exact role in pathomechanism of autism.

A glassy carbon electrode modified with a composite consisting of gold nanoparticle, reduced graphene oxide and poly(L-arginine) for simultaneous voltammetric determination of dopamine, serotonin and L-tryptophan.

A glassy carbon electrode (GCE) was modified with poly(L-arginine) (P-Arg), reduced graphene oxide (rGO) and gold nanoparticle (AuNP) to obtain an electrode for simultaneous determination of dopamine (DA), serotonin (5-HT) and L-tryptophan (L-Trp) in the presence of ascorbic acid (AA). The modified GCE was prepared via subsequent 'layer-by-layer' deposition using an electrochemical technique. The surface morphology of the modified electrode was studied by scanning electron microscopy, and electrochemical characterizations were carried out via cyclic voltammetry and electrochemical impedance spectroscopy. The modified electrode showed excellent electrocatalytic activity toward DA, 5-HT and L-Trp at pH 7.0. Figures of merit for the differential pulse voltammetric reponse are as follows: (a) Response to DA is linear in two intervals, viz. 1.0-50 nM and 1.0-50 µM DA concentration range, the typical working voltage is 202 mV (vs. Ag/AgCl), and the detection limit is 1 nM (at an S/N ratio of 3). For 5-HT, the respective data are 10 to 500 nM and 1.0 to 10 µM, 381 mV, and 30 nM. For L-Trp, the respective data are 10-70 nM and 10-100 µM, 719 mV, and 0.1 µM. The modified GCE is fairly selective. It was successfully applied to the simultaneous determination of DA, 5-HT, and L-Trp in spiked urine samples, and high recovery rates were found. Graphical abstract Schematic presentation of the voltammetric sensor based on a glassy carbon electrode modified with poly(L-arginine), reduced graphene oxide (rGO) and gold nanoparticle (GCE/P-Arg/ErGO/AuNP) for simultaneous determination of dopamine (DA), serotonin (5-HT) and L-tryptophan (L-Trp).

Differentiating Medicated Patients Suffering from Major Depressive Disorder from Healthy Controls by Spot Urine Measurement of Monoamines and Steroid Hormones.

Introduction: Major Depressive Disorder (MDD) is a common psychiatric disorder. Currently, there is no objective, cost-effective and non-invasive method to measure biological markers related to the pathogenesis of MDD. Previous studies primarily focused on urinary metabolite markers which are not proximal to the pathogenesis of MDD. Herein, we compare urinary monoamines, steroid hormones and the derived ratios amongst MDD when compared to healthy controls. Methods: Morning urine samples of medicated patients suffering from MDD (n = 47) and healthy controls (n = 41) were collected. Enzyme-linked immunosorbent assay (ELISA) was performed to measure five biomarkers: cortisol, dopamine, noradrenaline, serotonin and sulphate derivative of dehydroepiandrosterone (DHEAS). The mean urinary levels and derived ratios of monoamines and steroid hormones were compared between patients and controls to identify potential biomarkers. The receiver operative characteristic curve (ROC) analysis was conducted to evaluate the diagnostic performance of potential biomarkers. Results: Medicated patients with MDD showed significantly higher spot urine ratio of DHEAS/serotonin (1.56 vs. 1.19, p = 0.004) and lower ratio of serotonin/dopamine (599.71 vs. 888.60, p = 0.008) than healthy controls. A spot urine serotonin/dopamine ratio cut-off of >667.38 had a sensitivity of 73.2% and specificity of 51.1%. Conclusions: Our results suggest that spot urine serotonin/dopamine ratio can be used as an objective diagnostic method for adults with MDD.

A review on electrochemical detection of serotonin based on surface modified electrodes.

Serotonin is one of the important neurotransmitters of our body. It's abnormal concentration is associated with multiple disorders and diseases. Sensitive and precise electrochemical determination of serotonin is not possible with bare working electrodes due to various reasons viz. electro-chemical fouling, presence of other biological molecules having similar oxidation potential, and lower concentration of serotonin in biological samples. Surface modification of working electrode is required for fast, precise, selective, and sensitive detection of serotonin. We have extensively reviewed the research approaches where serotonin has been sensitively detected using surface modified electrodes in the presence of other interfering agents. This review aims at presenting the electrochemical detection of serotonin using various surface modified electrodes such as glassy carbon, graphite, carbon fiber, diamond, screen printed, ITO, and metal electrodes modified with conducting polymers and polyelectrolytes, carbon nanomaterials, metal or metal oxide nanoparticles, biological compounds, and other conducting materials. The analytical figures of merits of various research approaches for detection of serotonin have been compared in the article. The properties of material used for surface modification, chemical interactions at the interfaces, and electrocatalytic effects of modified surfaces on sensing of serotonin have been thoroughly discussed in this review.

Two-Dimensional Capillary Electrophoresis with On-Line Sample Preparation and Cyclodextrin Separation Environment for Direct Determination of Serotonin in Human Urine.

An advanced two-dimensional capillary electrophoresis method, based on on-line combination of capillary isotachophoresis and capillary zone electrophoresis with cyclodextrin additive in background electrolyte, was developed for effective determination of serotonin in human urine. Hydrodynamically closed separation system and large bore capillaries (300-800 µm) were chosen for the possibility to enhance the sample load capacity, and, by that, to decrease limit of detection. Isotachophoresis served for the sample preseparation, defined elimination of sample matrix constituents (sample clean up), and preconcentration of the analyte. Cyclodextrin separation environment enhanced separation selectivity of capillary zone electrophoresis. In this way, serotonin could be successfully separated from the rest of the sample matrix constituents migrating in capillary zone electrophoresis step so that human urine could be directly (i.e., without any external sample preparation) injected into the analyzer. The proposed method was successfully validated, showing favorable parameters of sensitivity (limit of detection for serotonin was 2.32 ng·mL-1), linearity (regression coefficient higher than 0.99), precision (repeatability of the migration time and peak area were in the range of 0.02-1.17% and 5.25-7.88%, respectively), and recovery (ranging in the interval of 90.0-93.6%). The developed method was applied for the assay of the human urine samples obtained from healthy volunteers. The determined concentrations of serotonin in such samples were in the range of 12.4-491.2 ng·mL-1 that was in good agreement with literature data. This advanced method represents a highly effective, reliable, and low-cost alternative for the routine determination of serotonin as a biomarker in human urine.

Neuroanatomic pathways associated with monoaminergic dysregulation after stroke.

OBJECTIVE: We examined the complex relationship between lesion location, symptoms of depression (affective and apathetic), and monoamine dysfunction after stroke. METHODS: Magnetic resonance imaging was performed on 48 post-stroke patients that had been assessed for affective and apathetic symptoms using the Hospital Anxiety and Depression Scale and the Apathy Scale, respectively. Noradrenalin (NA), dopamine (DA), their metabolites, and a metabolite of serotonin (5-HT) were measured using 24-h urine samples, and 5-HT and 3-methoxy-4-hydroxyphenylglycol were measured using blood samples. We developed a statistical parametric map that displayed the associations between lesion location and both positive and negative alterations of monoamines and their metabolites. RESULTS: Multivariate analysis indicated that basal ganglia lesions and 5-HT showed relationships with affective symptoms, whereas homovanillic acid was related to apathetic symptoms. Univariate analysis showed no such relationships. However, decreases in NA and DA and increases in NA and DA turnover were related to lesions in the brainstem, whereas increases in NA and DA as well as decreases in NA and DA turnover were related to cortical and/or striatum lesions. 5-HT turnover data showed a pattern opposite to that seen for NA and DA turnover. CONCLUSIONS: Monoaminergic neuronal pathways are controlled by both receptor-mediated feedback mechanisms and turnover; thus, depletion of monoamines is not the only cause of depression and apathy. Moreover, the monoamine neuronal network might be divided into two branches, catecholamine (NA and DA) and 5-HT, both of which are anatomically and functionally interconnected and could respectively influence apathetic and affective symptoms of depression.

Nonylphenol Toxicity Evaluation and Discovery of Biomarkers in Rat Urine by a Metabolomics Strategy through HPLC-QTOF-MS.

Nonylphenol (NP) was quantified using liquid chromatography tandem mass spectrometry (LC-MS/MS) in the urine and plasma of rats treated with 0, 50, and 250 mg/kg/day of NP for four consecutive days. A urinary metabolomic strategy was originally implemented by high performance liquid chromatography time of flight mass spectrometry (HPLC-QTOF-MS) to explore the toxicological effects of NP and determine the overall alterations in the metabolite profiles so as to find potential biomarkers. It is essential to point out that from the observation, the metabolic data were clearly clustered and separated for the three groups. To further identify differentiated metabolites, multivariate analysis, including principal component analysis (PCA), orthogonal partial least-squares discriminant analysis (OPLS-DA), high-resolution MS/MS analysis, as well as searches of Metlin and Massbank databases, were conducted on a series of metabolites between the control and dose groups. Finally, five metabolites, including glycine, glycerophosphocholine, 5-hydroxytryptamine, malonaldehyde (showing an upward trend), and tryptophan (showing a downward trend), were identified as the potential urinary biomarkers of NP-induced toxicity. In order to validate the reliability of these potential biomarkers, an independent validation was performed by using the multiple reaction monitoring (MRM)-based targeted approach. The oxidative stress reflected by urinary 8-oxo-deoxyguanosine (8-oxodG) levels was elevated in individuals highly exposed to NP, supporting the hypothesis that mitochondrial dysfunction was a result of xenoestrogen accumulation. This study reveals a promising approach to find biomarkers to assist researchers in monitoring NP.