Dopamine detection
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Accumulation of misfolded α-synuclein (α-Syn) leads to the formation of Lewy bodies and is a major hallmark of Parkinson's disease (PD). The accumulation of α-Syn involves several post-translational modifications. Recently, though, glycation of α-Syn (advanced glycation end products) and activation of the receptor for advanced glycation end products (RAGE) have been linked to neuroinflammation, which leads to oxidative stress and accumulation of α-Syn. The present study aims to detect the effect of glycated α-Syn (gly-α-Syn)-induced synucleinopathy and loss of dopaminergic (DAergic) neurons in the development of PD. We isolated, purified, and prepared glycated recombinant human α-Syn using d-ribose. Gly-α-Syn was characterized by SDS-PAGE, intact mass analysis, and bottom-up peptide sequence through LC-HRMS/MS. The aggregation propensity of gly-α-Syn has been verified by morphological and shape analysis through Bio-AFM. The gly-α-Syn (2 μg/μL) was injected stereotaxically in the substantia nigra (SN) of ICR mice (3-4 months) and compared with the normal α-Syn, d ribose, and Tris-HCl/artificial CSF groups. 56 days postsurgery (DPS), an immunohistochemical examination was conducted to investigate gly-α-Syn-induced α-Syn accumulation, neuroinflammation, and neurodegeneration. The glycation of α-Syn led to the expression of transglutaminase 2 (TGM2), an enzyme that cross-linked with AGEs and may have caused the accumulation of α-Syn. Significant RAGE activation was also observed in gly-α-Syn, which might have induced glial cell activation, resulting in oxidative stress and, ultimately, apoptosis of dopaminergic neurons. It is important to note that TGM2, phosphorylated α-Syn, RAGE expression, and glial cell activation were only found in the gly-α-Syn group and not in the other groups. This suggests that gly-α-Syn plays a major role in synucleinopathy, neuroinflammation, and neurodegeneration. Overall, the present study demonstrated glycation of α-Syn as one of the important age-associated post-translational modifications that are involved in the degeneration of dopaminergic neurons, at least in a subset of the diabetic patients susceptible to developing PD.
- MeSH
- alfa-synuklein * metabolismus MeSH
- dopaminergní neurony * metabolismus patologie účinky léků MeSH
- glykosylace MeSH
- lidé MeSH
- myši inbrední ICR MeSH
- myši MeSH
- neuroglie * metabolismus patologie účinky léků MeSH
- oxidační stres MeSH
- Parkinsonova nemoc * metabolismus patologie MeSH
- produkty pokročilé glykace metabolismus MeSH
- receptor pro konečné produkty pokročilé glykace metabolismus MeSH
- substantia nigra metabolismus patologie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Úvodom: Nové poznatky sa premietli i do zmien klasifikácie ICD-11 pri klinických prejavoch duševnej poruchy súvisiacej s hraním hazardných hier. Na rozdiel od patologického hrania v ICD-10 už nie je možné stanoviť samostatnú diagnózu poruchy hazardného hrania, ak začiatok narušenia kontroly nad hazardným hraním je korešpondujúci s užívaním metamfetamínov. Cieľ štúdie: Preskúmať výskyt hazardného, resp. patologického hrania podľa ICD-10 u užívateľov so závislosťou od metamfetamínov v liečbe, a na vybraných mikro-ilustráciách poukázať aplikáciou nových kritérií podľa ICD-11 na potrebu revízie diagnóz. Pacienti a metódy: Šlo o retrospektívnu, deskriptívnu, klinickú štúdiu záznamov súboru 178 pacientov liečených pre závislosť od metamfetamínov doplnenú o tri vybrané mikro-ilustrácie umožňujúce poukázať na diferenciálnu diagnostiku podľa ICD-11 a jej patognomický význam. Výsledky: Hazardné hranie bolo zistené u 30 % pacientov liečených pre závislosť od metamfetamínov a u 18 % bola zároveň zistená diagnóza patologického hrania podľa ICD-10. Kvalitatívnou metódou, vybranými mikro-ilustráciami bola dokumentovaná zmena a nový spôsob diferenciálnej diagnostiky podľa ICD-11. Diskusia a záver: Zistenia štúdie preukázali vysoký výskyt narušenia kontroly ovládania impulzov pri hazardnom hraní u užívateľov so závislosťou od metamfeta-mínov, čo má u veľkej časti neurobiologické vysvetlenie mechanizmom priameho účinku dopamínu v CNS, obdobne ako pri komplikácii liečby agonistami dopamínu pri Parkinsonovej chorobe. V klasifikácii ICD-11 nejde o samostatnú diagnózu, len o možný behaviorálny príznak užívania psychoaktívnej látky. V závere autori rozoberajú dôsledky diagnostickej zmeny a rad potrebných zmien v klinickej praxi po adaptácii ICD-11.
Introduction: New knowledge was projected into the ICD-11 classification in clinical manifestation of mental disorder associated with gambling. The independent diagnostic category of gambling disorder according to ICD-11 should not be assessed, contrary to pathological gambling in ICD-10, if the onset of impaired control over gambling behaviour is corresponding to use of the methamphetamines. The aim of the study: To detect the prevalence of gambling/pathological gambling according to ICD-10 classification among methamphetamine users with dependence in treatment, and with selected micro case-illustrations indicate the necessity to revise the diagnosis by the implementation of new ICD-11 criteria. Patients and methods: It was retrospective, descriptive, clinical study of the medical records of 178 patients treated for methamphetamine dependence with the addition of three selected micro case-illustrations, which provided the opportunity to present different diagnostic assessment according to ICD-11 and its patognomic significance. Results: Gambling behaviour was detected in 30% of the patients treated for dependence on methamphetamines, in the same time 18% had the the diagnosis of pathological gambling according to ICD-10. The new diagnostic approach according to ICD-11 was documented with use of qualitative method by three micro case-illustrations. Discussion and conclusion: The findings of the study have shown the high prevalence of the impaired control over gambling behaviour among the patients in treatment due to dependence on methamphetamines, which in large part of them, had the neurobiological base due to the direct effects of dopamine on the CNS, so as it is sometimes in the treatment of Parkinson disease by dopamine agonists. According to ICD-11 it is not an independent diagnostic category, but it is only behavioural sign induced by psychoactive substance. Finally, the authors are discussing the consequences of the change in diagnostic assessment and the number of inevitable changes in the clinical practice after the adoption of ICD-11.
Accumulation of alpha-synuclein (α-syn) is central to the pathogenesis of Parkinson's disease (PD). Previous studies suggest that α-syn pathology may originate from the olfactory bulb (OB) or gut in response to an unknown pathogen and later progress to the different brain regions. Aging is viewed as the utmost threat to PD development. Therefore, studies depicting the role of age in α-syn accumulation and its progression in PD are important. In the present study, we gave intranasal rotenone microemulsion for 6 weeks in 12-month-old female BALB/c mice and found olfactory dysfunction after 4 and 6 weeks of rotenone administration. Interestingly, motor impairment was observed only after 6 weeks. The animals were sacrificed after 6 weeks to perform western blotting and immunohistochemical studies to detect α-syn pathology, neuroinflammation and neurodegeneration. We found α-syn accumulation in OB, striatum, substantia nigra (SN) and cortex. Importantly, we found significant glial cell activation and neurodegeneration in all the analysed regions which were absent in our previous published studies with 3 months old mice even after they were exposed to rotenone for 9 weeks indicating age is a crucial factor for α-syn induced neuroinflammation and neurodegeneration. We also observed increased iron accumulation in SN of rotenone-exposed aged mice. Moreover, inflammaging was observed in OB and striatum of 12-month-old BALB/c mice as compared to 3-month-old BALB/c mice. In conclusion, there is a difference in sensitivity between adult and aged mice in the development and progression of α-syn pathology and subsequent neurodegeneration, for which inflammaging might be the crucial probable mechanism.
- MeSH
- alfa-synuklein * metabolismus MeSH
- dopamin MeSH
- dopaminergní neurony metabolismus MeSH
- modely nemocí na zvířatech MeSH
- mozek metabolismus MeSH
- myši MeSH
- neurozánětlivé nemoci MeSH
- Parkinsonova nemoc * patologie MeSH
- rotenon toxicita MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
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
Prolaktinomy jsou nejčastější hormonálně aktivní adenomy hypofýzy. U žen je obvyklým příznakem onemocnění amenorea či jiná porucha menstruace. Diagnostika je založena na zjištění hyperprolaktinemie s verifikací adenomu v oblasti tureckého sedla pomocí magnetické rezonance. Ve většině případů má dobrý efekt samostatná farmakoterapie kabergolinem. Léčbu makroprolaktinomů je nutné individuálně zvažovat, uplatňuje se kombinace více modalit, například operace nebo ozáření gama nožem následované farmakoterapií.
Prolactinomas are the most common hormonally active pituitary adenomas. For women, amenorrhea or other menstrual disturbances are common symptoms of the disease. Diagnosis is based on the detection of hyperprolactinaemia with verification of the adenoma in the region of the Turkish saddle using magnetic resonance imaging. In most cases, cabergoline alone has a good effect. The treatment of macroprolactinomas must be individually considered; a combination of several modalities, such as surgery or gamma-knife irradiation followed by drug therapy, is applied.
- MeSH
- agonisté dopaminu aplikace a dávkování farmakologie terapeutické užití MeSH
- amenorea MeSH
- galaktorea MeSH
- hyperprolaktinemie diagnóza etiologie MeSH
- kabergolin aplikace a dávkování farmakologie terapeutické užití MeSH
- léková rezistence MeSH
- lidé MeSH
- magnetická rezonanční tomografie MeSH
- mladý dospělý MeSH
- nádory hypofýzy chirurgie diagnóza farmakoterapie MeSH
- neúspěšná terapie MeSH
- prolaktin analýza krev účinky léků MeSH
- prolaktinom * chirurgie diagnóza farmakoterapie MeSH
- radiochirurgie MeSH
- Check Tag
- lidé MeSH
- mladý dospělý MeSH
- ženské pohlaví MeSH
The here presented work is focused on the development of a method for detection of microbial contamination of food based on uracil-selective synthetic receptors. Because uracil may serve as an indicator of bacterial contamination, its selective and on-site detection may prevent spreading of foodborne diseases. The synthetic receptors were created by molecular imprinting. Molecularly imprinted polymers for selective uracil isolation were prepared by a non-covalent imprinting method using dopamine as a functional monomer. Detection of isolated uracil was performed by capillary electrophoresis with absorption detection (λ - 260 nm). The conditions of preparation of molecularly imprinted polymers, their binding properties, adsorption kinetics and selectivity were investigated in detail. Furthermore, the prepared polymer materials were used for selective isolation and detection of uracil from complex samples as tomato products by miniaturized electrophoretic system suggesting the potential of in situ analysis of real samples.
- MeSH
- adsorpce MeSH
- molekulový imprinting * MeSH
- polymery MeSH
- receptory umělé * MeSH
- uracil MeSH
- Publikační typ
- časopisecké články MeSH
Novel porous boron-doped diamond (BDDporous)-based materials have attracted lots of research interest due to their enhanced detection ability and biocompatibility, favouring them for use in neuroscience. This study reports on morphological, spectral, and electrochemical characterisation of three BDDporous electrodes of different thickness given by a number of deposited layers (2, 3 and 5). These were prepared using microwave plasma-enhanced chemical vapour deposition on SiO2 nanofiber-based scaffolds. Further, the effect of number of layers and poly-l-lysine coating, commonly employed in neuron cultivation experiments, on sensing properties of the neurotransmitter dopamine in a pH 7.4 phosphate buffer media was investigated. The boron doping level of ∼2 × 1021 atoms cm-3 and increased content of non-diamond (sp2) carbon in electrodes with more layers was evaluated by Raman spectroscopy. Cyclic voltammetric experiments revealed reduced working potential windows (from 2.4 V to 2.2 V), higher double-layer capacitance values (from 405 μF cm-2 to 1060 μF cm-2), enhanced rates of electron transfer kinetics and larger effective surface areas (from 5.04 mm2 to 7.72 mm2), when the number of porous layers increases. For dopamine, a significant boost in analytical performance was recognized with increasing number of layers using square-wave voltammetry: the highest sensitivity of 574.1 μA μmol-1 L was achieved on a BDDporous electrode with five layers and dropped to 35.9 μA μmol-1 L when the number of layers decreased to two. Consequently, the lowest detection limit of 0.20 μmol L-1 was obtained on a BDDporous electrode with five layers. Moreover, on porous electrodes, enhanced selectivity for dopamine detection in the presence of ascorbic acid and uric acid was demonstrated. The application of poly-l-lysine coating on porous electrode surface resulted in a decrease in dopamine peak currents by 17% and 60% for modification times of 1 h and 15 h, respectively. Hence, both examined parameters, the number of deposited porous layers and the presence of poly-l-lysine coating, were proved to considerably affect the characteristics and performance of BDDporous electrodes.
Clinical diagnosis of Parkinson's disease (PD) occurs typically when a substantial proportion of dopaminergic neurons in the substantia nigra (SN) already died, and the first motor symptoms appear. Therefore, tools enabling the early diagnosis of PD are essential to identify early-stage PD patients in which neuroprotective treatments could have a significant impact. Here, we test the utility and sensitivity of the diffusion kurtosis imaging (DKI) in detecting progressive microstructural changes in several brain regions of mice exposed to chronic intragastric administration of rotenone, a mouse model that mimics the spatiotemporal progression of PD-like pathology from the ENS to the SN as described by Braak's staging. Our results show that DKI, especially kurtosis, can detect the progression of pathology-associated changes throughout the CNS. Increases in mean kurtosis were first observed in the dorsal motor nucleus of the vagus (DMV) after 2 months of exposure to rotenone and before the loss of dopaminergic neurons in the SN occurred. Remarkably, we also show that limited exposure to rotenone for 2 months is enough to trigger the progression of the disease in the absence of the environmental toxin, thus suggesting that once the first pathological changes in one region appear, they can self-perpetuate and progress within the CNS. Overall, our results show that DKI can be a useful radiological marker for the early detection and monitoring of PD pathology progression in patients with the potential to improve the clinical diagnosis and the development of neuroprotective treatments.
- MeSH
- aplikace orální MeSH
- bludiště - učení účinky léků fyziologie MeSH
- časové faktory MeSH
- dopaminergní neurony účinky léků patologie MeSH
- insekticidy toxicita MeSH
- myši inbrední C57BL MeSH
- myši MeSH
- parkinsonské poruchy chemicky indukované diagnostické zobrazování patologie MeSH
- progrese nemoci * MeSH
- rotenon aplikace a dávkování toxicita MeSH
- zobrazování difuzních tenzorů metody MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
OBJECTIVE: To determine whether restricting the use of inotrope after diagnosis of low blood pressure (BP) in the first 72 hours of life affects survival without significant brain injury at 36 weeks of postmenstrual age (PMA) in infants born before 28 weeks of gestation. DESIGN: Double-blind, placebo-controlled randomised trial. Caregivers were masked to group assignment. SETTING: 10 sites across Europe and Canada. PARTICIPANTS: Infants born before 28 weeks of gestation were eligible if they had an invasive mean BP less than their gestational age that persisted for ≥15 min in the first 72 hours of life and a cerebral ultrasound free of significant (≥ grade 3) intraventricular haemorrhage. INTERVENTION: Participants were randomly assigned to saline bolus followed by either a dopamine infusion (standard management) or placebo (5% dextrose) infusion (restrictive management). PRIMARY OUTCOME: Survival to 36 weeks of PMA without severe brain injury. RESULTS: The trial terminated early due to significant enrolment issues (7.7% of planned recruitment). 58 infants were enrolled between February 2015 and September 2017. The two groups were well matched for baseline variables. In the standard group, 18/29 (62%) achieved the primary outcome compared with 20/29 (69%) in the restrictive group (p=0.58). Additional treatments for low BP were used less frequently in the standard arm (11/29 (38%) vs 19/29 (66%), p=0.038). CONCLUSION: Though this study lacked power, we did not detect major differences in clinical outcomes between standard or restrictive approach to treatment. These results will inform future studies in this area. TRIAL REGISTRATION NUMBER: NCT01482559, EudraCT 2010-023988-17.
- MeSH
- dopamin aplikace a dávkování škodlivé účinky terapeutické užití MeSH
- dvojitá slepá metoda MeSH
- gestační stáří MeSH
- hypotenze farmakoterapie mortalita MeSH
- kardiotonika aplikace a dávkování škodlivé účinky terapeutické užití MeSH
- lidé MeSH
- novorozenci extrémně nezralí * MeSH
- novorozenec MeSH
- poranění mozku chemicky indukované MeSH
- Check Tag
- lidé MeSH
- novorozenec MeSH
- Publikační typ
- časopisecké články MeSH
- multicentrická studie MeSH
- randomizované kontrolované studie MeSH
A device with four parallel channels was designed and manufactured by 3D printing in titanium. A simple experimental setup allowed splitting of the mobile phase in four parallel streams, such that a single sample could be analysed four times simultaneously. The four capillary channels were filled with a monolithic stationary phase, prepared using a zwitterionic functional monomer in combination with various dimethacrylate cross-linkers. The resulting stationary phases were applicable in both reversed-phase and hydrophilic-interaction retention mechanisms. The mobile-phase composition was optimized by means of a window diagram so as to obtain the highest possible resolution of dopamine precursors and metabolites on all columns. Miniaturized electrochemical detectors with carbon fibres as working electrodes and silver micro-wires as reference electrodes were integrated in the device at the end of each column. Experimental separations were successfully compared with those predicted by a three-parameter retention model. Finally, dopamine was determined in human urine to further confirm applicability of the developed device.
