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DOPA
Terms modified by DOPA Selected AbstractsDifferential effect of dopamine on mitosis in early postnatal albino and pigmented rat retinaeDEVELOPMENTAL NEUROBIOLOGY, Issue 1 2006Ines Kralj-Hans Abstract Insufficient levels of L -DOPA, released from the retinal pigment epithelium (RPE), in albino animals are considered responsible for the abnormal development of the underlying neural retina. L -DOPA normalizes retinal neurogenesis by reducing levels of cell proliferation either by acting on the cells directly or by being converted into dopamine. Here we report the effects of dopamine on mitosis in early postnatal neural retinae from albino and pigmented rats, using 4D (x, y, z and time) confocal microscopy. Exogenous dopamine significantly prolongs mitosis in retinae from albino, but not pigmented, animals. As fewer cells move into and divide in the ventricular zone (VZ) in the presence of dopamine, we conclude that the overall cell cycle is affected. The D1 receptor blocker, SCH 23390, inhibits these effects. Thus, the differential effects of dopamine on neural retinae from pigmented and albino rats in vitro must result from the activation of D1 receptors, which are present in the retina from birth. Immunohistochemical labeling of D1 receptors shows that the pattern of their distribution is similar between pigmentation phenotypes, but levels of expression may be elevated in albinos. Labeling is most intense in the inner plexiform layer but is present throughout the neuroblastic layer. These findings are discussed in light of previous reports of reduced catecholamine levels in the albino retina. © 2005 Wiley Periodicals, Inc. J Neurobiol, 2006 [source] Dopamine and sensory tissue development in Drosophila melanogasterDEVELOPMENTAL NEUROBIOLOGY, Issue 4 2001Wendi Neckameyer Abstract Dopamine is an important signaling molecule in the nervous system; it also plays a vital role in the development of diverse non-neuronal tissues in the fruit fly Drosophila melanogaster. The current study demonstrates that males depleted of dopamine as third instar larvae (via inhibition of the biosynthetic enzyme tyrosine hydroxylase) demonstrated abnormalities in courtship behavior as adults. These defects were suggestive of abnormalities in sensory perception and/or processing. Electroretinograms (ERGs) of eyes from adults depleted of dopamine for 1 day as third instar larvae revealed diminished or absent on- and off-transients. These sensory defects were rescued by the addition of L -DOPA in conjunction with tyrosine hydroxylase inhibition during the larval stage. Depletion of dopamine in the first or second larval instar was lethal, but this was not due to a general inhibition of proliferative cells. To establish that dopamine was synthesized in tissues destined to become part of the adult sensory apparatus, transgenic lines were generated containing 1 or 4 kb of 5, upstream sequences from the Drosophila tyrosine hydroxylase gene (DTH) fused to the E. coli ,-galactosidase reporter. The DTH promoters directed expression of the reporter gene in discrete and consistent patterns within the imaginal discs, in addition to the expected expression in gonadal, brain, and cuticular tissues. The ,-galactosidase expression colocalized with tyrosine hydroxylase protein. These results are consistent with a developmental requirement for dopamine in the normal physiology of adult sensory tissues. © 2001 John Wiley & Sons, Inc. J Neurobiol 47: 280,294, 2001 [source] Spectroelectrochemical and Voltammetric Studies of L -DOPAELECTROANALYSIS, Issue 2 2003Xiaoqiang Liu Abstract The electrooxidation of L -dopa at GC electrode was studied by in situ UV-vis spectroelectrochemistry (SEC) and cyclic voltammetry. The mechanism of electrooxidation and some reaction parameters were obtained. The results showed that the whole electrooxidation reaction of L -dopa at glassy carbon (GC) electrode was an irreversible electrochemical process followed by a chemical reaction in neutral solution (EC mechanism). The spectroelectrochemical data were treated by the double logarithm method together with nonlinear regression, from which the formal potential E0=228,mV, the apparent electron-transfer number of the electrooxidation reaction ,n=0.376 (R=0.99, SD=0.26), the standard electrochemical rate constant k0=(3.93±0.12)×10,4,cm s,1 (SD=1.02×10,2), and the formation equilibrium constant of the following chemical reaction kc=(5.38±0.34)×10,1,s,1 (SD=1.02×10,2) were also obtained. [source] Microchip capillary electrophoresis with a cellulose-DNA-modified screen-printed electrode for the analysis of neurotransmittersELECTROPHORESIS, Issue 15 2005Muhammad Johirul Abstract A microfluidic chip based on capillary electrophoresis coupled with a cellulose-single-stranded DNA (cellulose-ssDNA) modified electrode was used for the simultaneous analysis of dopamine (DA), norepinephrine (NE), 3,4-dihydroxy- L -phenylalanine (L -DOPA), 3,4-dihydroxyphenylacetic acid (DOPAC), and ascorbic acid (AA). The modification of the electrode improved the electrophoretic analysis performance by lowering the detection potential and enhancing the signal-to-noise characteristic without surface poisoning of the electrode. The sensitivity of the modified electrode was about 12 times higher than those of the bare ones. The test compounds were separated using a 62,mm long separation channel at the separation field strength of +200,V/cm within 220,s in a 10,mM phosphate buffer (pH,7.4). The most favorable potential for the amperometric detection was 0.7,V (vs. Ag/AgCl). A reproducible response (relative standard deviation of 1.3, 1.3, 2.1, 3.1, 3.4% for DA, NE, L -DOPA, DOPAC, and AA, respectively, for n,=,9) for repetitive sample injections reflected the negligible electrode fouling at the cellulose-ssDNA modified electrode. Square-wave voltammetric analyses reflected the sensitivities of the modified electrode for DA, NE, L -DOPA, DOPAC, and AA which were 1.78, 0.82, 0.69, 2.45, and 1.23,nC/µM with detection limits of 0.032, 0.93, 1.13, 0.31, and 0.62,µM, respectively. The applicability of this microsystem to real sample analysis was demonstrated. [source] ISTA13-catecholamine toxicity and metabolism in the ciliated protozoan, Tetrahymena pyriformisENVIRONMENTAL TOXICOLOGY, Issue 6 2009Asad Ud-Daula Abstract A high throughput culture methodology of unicellular eukaryote Tetrahymena pyriformis, strain GL were used for the determination of catecholamines toxicity and their metabolism. Catecholamines exhibited acute toxicity to Tetrahymena cells where dopamine and L -DOPA showed higher toxic potential of EC10 (0.39 and 0.63 mg/L, respectively) and EC20 (1.1 and 1.0 mg/L, respectively). All the testing catecholamines were highly degradable in the PPY-medium due to the oxidizing environment during incubation. They were also naturally synthesized and released by Tetrahymena cells into the culture medium and increasingly accumulated with time where as noradrenalin demonstrated significant results. Cells were exposed with physiological concentration (0.12 mg/L) and one higher concentration (8.0 mg/L) of catecholamines, resulting noradrenalin depletion and in vivo generation of a metabolite in response to dopamine with higher concentration treatment. This dopamine metabolite was relatively nonpolar compared with the catecholamines and was eluted later from the reverse phase C-18 column. © 2008 Wiley Periodicals, Inc. Environ Toxicol, 2009. [source] Systemic concentrations of antioxidants and biomarkers of macromolecular oxidative damage in horses with grass sicknessEQUINE VETERINARY JOURNAL, Issue 2 2003B. C. McGORUM Summary Reasons for performing study: The aetiopathogenesis of equine grass sickness (EGS) is unknown. The role of free radical-mediated neuronal damage has not previously been investigated in this condition. Objectives: To investigate the potential contribution of oxidative damage and antioxidant status to neurodegeneration in EGS. Methods: Systemic levels of surrogate biomarkers were determined in 10 horses with acute EGS and in 2 control populations; 10 healthy horses co-grazing with the 10 EGS horses at the onset of clinical disease, and 10 healthy mares grazing where EGS has not been reported. Results: EGS horses had alterations in levels of several antioxidants, consistent with oxidative stress, the acute phase response and/or the secondary metabolic complications of EGS. EGS horses had elevated plasma dihydroxyphenylalanine (DOPA) levels. Conclusions: The elevated DOPA levels probably reflected a generalised disturbance of catecholamine metabolism rather than increased DOPA production via free radical-mediated oxidation of tyrosine. However, there was no evidence of systemic macromolecular oxidative damage. Potential clinical relevance: Further work is required to determine whether macromolecular oxidative damage occurring at the neuronal level contributes to EGS. [source] Role of dopachrome conversion enzyme in the melanization of filarial worms in mosquitoesINSECT MOLECULAR BIOLOGY, Issue 6 2005C.-Y. Huang Abstract Melanization is an effective defence reaction of mosquito hosts against invading parasites. In mosquitoes, the biosynthesis of melanin is initiated by the hydroxylation of tyrosine to DOPA by phenoloxidase (PO). DOPA is a branch point of the melanization reaction; it may be oxidized to dopaquinone by PO or be decarboxylated to dopamine by dopa decarboxylase. Further oxidation of dopaquinone by PO produces dopachrome. Dopachrome is then converted to 5, 6-dihydroxyindole by dopachrome conversion enzyme (DCE) to produce melanin. The conversion of dopachrome is a rate-limiting step of the melanization reaction, and the presence of PO and DCE significantly accelerates melanization reactions. In this study, a cDNA encoding DCE was cloned from the mosquito Armigeres subalbatus. Real-time PCR analysis revealed increased transcripts from haemocytes in microfilariae (mf)-inoculated mosquitoes. Gene silencing using double-stranded RNA was used to elucidate the role of DCE in the melanization reaction of parasites in Ar. subalbatus. The levels of both DCE transcripts and protein in gene knockdown mosquitoes were dramatically reduced. Compared with controls, the degree of melanization of mf in DCE-knockdown mosquitoes was significantly decreased. These results suggest that DCE is a critical enzyme that is required for effective melanization immune responses. [source] Trifluoromethanesulfonic acid, an alternative solvent medium for the direct electrophilic fluorination of DOPA: new syntheses of 6-[18F]fluoro- L -DOPA and 6-[18F]fluoro- D -DOPAJOURNAL OF LABELLED COMPOUNDS AND RADIOPHARMACEUTICALS, Issue 14 2007Babak Behnam Azad Abstract Previous work from this laboratory has shown that the direct fluorination of 3, 4-dihydroxy-phenyl- L -alanine (L -DOPA) in anhydrous HF (aHF) or BF3/HF with F2 is an efficient method for the synthesis of 6-fluoro- L -DOPA. Since then, 18F-labeled 6-fluoro- L -DOPA ([18F]6-fluoro- L -DOPA) has been used to study presynaptic dopaminergic function in the human brain and to monitor gastrointestinal carcinoid tumors. This work demonstrates that the reactivity and selectivity of F2 toward L -DOPA in CF3SO3H is comparable with that in aHF. This new synthetic procedure has led to the production of [18F]fluoro- L -DOPA and [18F]fluoro-D-DOPA isomers in 17±2% radiochemical yields (decay corrected with respect to [18F]F2). The 2- and 6-FDOPA isomers were separated by HPLC and subsequently characterized by 19F NMR spectroscopy. The corresponding [18F]-FDOPA enantiomers have been obtained in clinically useful quantities by a synthetic approach that avoids the use of aHF. Copyright © 2007 John Wiley & Sons, Ltd. [source] Towards stereoselective radiosynthesis of ,-[11C]methyl-substituted aromatic ,-amino acids , a challenge of creation of quaternary asymmetric centre in a very short time,JOURNAL OF LABELLED COMPOUNDS AND RADIOPHARMACEUTICALS, Issue 5-6 2007Alexander Popkov Abstract In positron emission tomography (PET) , -methyl amino acids have two potential applications: As analogues of neutransmitter precursors for the study of neurodegenerative diseases; as non-metabolised analogues of proteinogenic amino acids for the study of amino acid uptake into normal and cancer cells. Clinical applications of such amino acids are strongly limited due to their poor availability. We carried out [11C]methylation of metalocomplex synthons derived from protected DOPA or tyrosine. For [11C]methylation, sodium hydroxide (5 mg of fine dry powder) was sealed in a vial, which was flushed with dry nitrogen before addition of a solution of the complex (10 mg) and 11CH3I in 1,3-dimethylimidazolidin-2-one (300 µl). After 10 min at 25°C, a 9% radiochemical yield (decay-corrected) of a mixture of the diastereomeric , -[11C]methylDOPA complexes or a 7% radiochemical yield of a mixture of the diastereomeric , -[11C]methyltyrosine complexes was achieved. Individual diastereomers were successfully separated by preparative HPLC, diluted with excess of water and extracted on C18 cartridges. Optimisation of the procedure including hydrolysis of the complexes (hydrolytic deprotection of enantiomerically pure amino acids) and subsequent purification of the enantiomers of , -[11C]methylDOPA and , -[11C]methyltyrosine is underway. Copyright © 2007 John Wiley & Sons, Ltd. [source] Increased dopamine and its metabolites in SH-SY5Y neuroblastoma cells that express tyrosinaseJOURNAL OF NEUROCHEMISTRY, Issue 2 2003Takafumi Hasegawa Abstract Oxidized metabolites of dopamine, known as dopamine quinone derivatives, are thought to play a pivotal role in the degeneration of dopaminergic neurons. Although such quinone derivatives are usually produced via the autoxidation of catecholamines, tyrosinase, which is a key enzyme in melanin biosynthesis via the production of DOPA and subsequent molecules, may potentially accelerate the induction of catecholamine quinone derivatives by its oxidase activity. In the present study, we developed neuronal cell lines in which the expression of human tyrosinase was inducible. Overexpression of tyrosinase in cultured cell lines resulted in (i) increased intracellular dopamine content; (ii) induction of oxidase activity not only for DOPA but also for dopamine; (iii) formation of melanin pigments in cell soma; and (iv) increased intracellular reactive oxygen species. Interestingly, the expressed tyrosinase protein was initially distributed in the entire cytoplasm and then accumulated to form catecholamine-positive granular structures by 3 days after the induction. The granular structures consisted of numerous rounded, dark bodies of melanin pigments and were largely coincident with the distribution of lysosomes. This cellular model that exhibits increased dopamine production will provide a useful tool for detailed analyses of the potentially noxious effects of oxidized catecholamine metabolites. [source] Cellular and behavioural effects of the adenosine A2a receptor antagonist KW-6002 in a rat model of l -DOPA-induced dyskinesiaJOURNAL OF NEUROCHEMISTRY, Issue 6 2003M. Lundblad Abstract We have examined the ability of KW-6002, an adenosine A2a antagonist, to modulate the dyskinetic effects of l -DOPA in 6-hydroxydopamine-lesioned rats. In animals rendered dyskinetic by a previous course of l -DOPA treatment, KW-6002 did not elicit any abnormal involuntary movements on its own, but failed to reduce the severity of dyskinesia when coadministered with l -DOPA. A second experiment was undertaken in order to study the effects of KW-6002 in l -DOPA-naive rats. Thirty-five animals were allotted to four groups to receive a 21-day treatment with: (i) KW-6002 (10 mg/kg/day); (ii) l -DOPA (6 mg/kg/day) i.p.; (iii) KW-6002 plus l -DOPA (same doses as above) or (iv) vehicle. Chronic treatment with KW-6002-only produced a significant relief of motor disability in the rotarod test in the absence of any abnormal involuntary movements. Combined treatment with l -DOPA and KW-6002 improved rotarod performance to a significantly higher degree than did each of the two drugs alone. However, this combined treatment induced dyskinesia to about the same degree as did l -DOPA alone. In situ hybridization histochemistry showed that KW-6002 treatment alone caused an approximately 20% reduction in the striatal levels of preproenkephalin mRNA, whereas neither the coadministration of KW-6002 and l -DOPA nor l -DOPA alone significantly altered the expression of this transcript in the dopamine-denervated striatum. Either alone or in combination with l -DOPA, KW-6002 did not have any modulatory effect on prodynorphin mRNA expression or FosB/,FosB-like immunoreactivity in the dopamine-denervated striatum. These results show that monotreatment with an adenosine A2a receptor antagonist can relieve motor disability without inducing behavioural and cellular signs of dyskinesia in rats with 6-hydroxydopamine lesions. Cotreatment with KW-6002 and l -DOPA potentiates the therapeutic effect but not the dyskinesiogenic potential of the latter drug. [source] Endogenously released DOPA is a causal factor for glutamate release and resultant delayed neuronal cell death by transient ischemia in rat striataJOURNAL OF NEUROCHEMISTRY, Issue 3 2001Nobuya Furukawa Glutamate is implicated in neuronal cell death. Exogenously applied DOPA by itself releases neuronal glutamate and causes neuronal cell death in in vitro striatal systems. Herein, we attempt to clarify whether endogenous DOPA is released by 10 min transient ischemia due to four-vessel occlusion during rat striatal microdialysis and, further, whether DOPA, when released, functions to cause glutamate release and resultant delayed neuronal cell death. Ischemia increased extracellular DOPA, dopamine, and glutamate, and elicited neuronal cell death 96 h after ischemic insult. Inhibition of striatal l -aromatic amino acid decarboxylase 10 min before ischemia increased markedly basal DOPA, tripled glutamate release with a tendency of decrease in dopamine release by ischemia, and exaggerated neuronal cell death. Intrastriatal perfusion of 10,30 nm DOPA cyclohexyl ester, a competitive DOPA antagonist, 10 min before ischemia, concentration-dependently decreased glutamate release without modification of dopamine release by ischemia. At 100 nm, the antagonist elicited a slight ceiling effect on decreases in glutamate release by ischemia and protected neurons from cell death. Glutamate was released concentration-dependently by intrastriatal perfusion of 0.3,1 mm DOPA and stereoselectively by 0.6 mm DOPA. The antagonist elicited no hypothermia during and after ischemia. Endogenously released DOPA is an upstream causal factor for glutamate release and resultant delayed neuronal cell death by brain ischemia in rat striata. DOPA antagonist has a neuroprotective action. [source] Endogenous melatonin protects L -DOPA from autoxidation in the striatal extracellular compartment of the freely moving rat: potential implication for long-term L -DOPA therapy in Parkinson's diseaseJOURNAL OF PINEAL RESEARCH, Issue 3 2006Gaia Rocchitta Abstract:, We previously showed, using microdialysis, that autoxidation of exogenous L-dihydroxyphenylalanine (l -DOPA) occurs in vivo in the extracellular compartment of the freely moving rat, with a consequent formation of l -DOPA semiquinone (l -DOPA-SQ). In the present study, intrastriatal infusion of l -DOPA (1.0 ,m for 200 min) increased dialysate l -DOPA concentrations (maximum increases up to 116-fold baseline values); moreover, l -DOPA-SQ was detected in dialysates. Individual dialysate concentrations of l -DOPA were negatively correlated with those of l -DOPA-SQ. Co-infusion of N -acetylcysteine (100 ,m) or melatonin (50 ,m) increased l -DOPA (up to 151- and 246-fold, respectively) and decreased l -DOPA-SQ (by about 53% and 87%, respectively) dialysate concentrations. Systemic l -DOPA [25 mg/kg intraperitoneally (i.p.) twice in a 12-h interval] significantly increased striatal baseline dialysate concentrations of l -DOPA and decreased dopamine (DA) and ascorbic acid (AsAc) concentrations, when compared with controls. Following systemic l -DOPA, l -DOPA-SQ was detected in dialysates. Endogenous melatonin was depleted in rats maintained on a 24-h light cycle for 1 wk. In melatonin-depleted rats, systemic l -DOPA induced a smaller increase in dialysate l -DOPA, a greater increase in l -DOPA-SQ formation, and a greater reduction in DA and AsAc dialysate concentrations. Co-administration of melatonin (5.0 mg/kg, i.p., twice in a 12-h interval) with l -DOPA, in control as well as in light-exposed rats, significantly increased dialysate l -DOPA concentrations, greatly inhibited l -DOPA-SQ formation, and restored up to the control values dialysate DA and AsAc concentrations. These findings demonstrate that endogenous melatonin protects exogenous l -DOPA from autoxidation in the extracellular compartment of the striatum of freely moving rats; moreover, systemic co-administration of melatonin with l -DOPA markedly increases striatal l -DOPA bioavailability in control as well as in melatonin-depleted rats. These results may be of relevance to the long-term l -DOPA therapy of Parkinson's disease. [source] The hair follicle melanocytes in vitiligo in relation to disease durationJOURNAL OF THE EUROPEAN ACADEMY OF DERMATOLOGY & VENEREOLOGY, Issue 8 2009T S Anbar Abstract Background and aims, Vitiligo is an acquired pigmentary disorder of skin and hair. Active melanocytes in hair follicles can be detected by DOPA and immunohistochemical staining, while amelanotic melanocytes can only be detected by the latter. None of the studies on hair melanocytes in vitiligo discussed the effect of disease duration on these melanocytes. Here, we study the presence of melanotic and amelanotic melanocytes in vitiligo hair follicles and statistically correlating their presence with the disease duration. Methods, This study was conducted on 30 patients with vitiligo and 10 normal volunteers. Three biopsies were taken from each patient: two from black and white hairs from vitiliginous areas and the third from apparently normal skin of the same patients. Sections were stained by DOPA reaction and NKI/beteb then examined for the presence of melanocytes. The presence of melanocytes and the disease duration were correlated statistically using the t -test. Results, Active melanocytes were detected in black hairs of 6.7% of vitiligo patients and in 100% of apparently normal skin of the same patients and controls. On examining black hairs of the 28 vitiligo patients with negative DOPA reaction, 19 of them (67.9%) showed positive NKI/beteb stain. Disease duration was inversely correlated with the melanocytes' presence within hair follicles. Melanocytes were absent from 100% of white hairs. Conclusions, The melanotic melanocytes were the first target of the disease process followed by the amelanotic melanocytes. Since the disappearance of the latter was inversely correlated with the disease duration, early treatment in vitiligo is advised. Conflicts of interest None declared. [source] Correlation between antiradical activity and stability of betanine from Beta vulgaris L roots under different pH, temperature and light conditionsJOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE, Issue 7 2001A Pedreño Abstract When the antiradical activity and stability of betanine were studied at pH values of 3.5 and 8.5 and temperatures of 25, 50 and 75,°C, the results showed that the antiradical activity was greater at acidic pH and lower at higher temperatures. At basic pH the activity of betanine correlated well with its stability at the three temperatures assayed, suggesting that the degradation products, betalamic acid (BA) and cyclo DOPA 5- O -,- D -glucoside (CDG), did not contribute to this activity under the experimental conditions used. However, at acidic pH the degradation product, CDG, did seem to contribute to the antiradical activity. Furthermore, at pH 3.5, betanine stability was so great that light conditions had no effect on the antiradical activity. At basic pH, too, light had no effect on betanine activity owing to the high instability of the pigment. © 2001 Society of Chemical Industry [source] Depression in multiple system atrophy: A case reportPSYCHIATRY AND CLINICAL NEUROSCIENCES, Issue 4 2000Kyoko Goto MD Abstract A 53-year-old woman who developed depression as the first symptom of multiple system atrophy was treated. Depression was followed successively by autonomic failure, parkinsonism and cerebellar ataxia. Treatment with L -DOPA, L -threo-DOPS, and thyroid releasing hormone was associated with improvement of autonomic failure and parkinsonism. As for depression, scores on the Zung scale and the Hamilton scale improved from 58 to 49 and from 30 to 22, respectively. This case is remarkable in that depression preceded neurologic dysfunction and was managed successfully by antiparkinsonian medication. A common underlying disturbance may be responsible for the depression and neurologic dysfunction in multiple system atrophy. [source] The Absence of Phosphorylated Tyrosine Hydroxylase Expression in the Purkinje Cells of the Ataxic Mutant Pogo MouseANATOMIA, HISTOLOGIA, EMBRYOLOGIA, Issue 3 2006N. S. Lee Summary The pogo mouse is a new ataxic autosomal recessive mutant that arose in Korean wild mice (KJR/Mskist). Its ataxic phenotype includes difficulty in maintaining a normal posture and the inability to walk in a straight line. Several studies have reported that tyrosine hydroxylase (TH) is persistently ectopically expressed in particular subsets of Purkinje cells in a parasagittal banding pattern in several ataxic mutant mice, e.g. tottering alleles and pogo mice. In this present study, we examined the expression of an enzymatically active form of TH and phosphorylated TH at Ser40 (phospho-TH) by using immunohistochemistry and double immunofluorescence in the cerebellum of pogo mice. TH immunostaining appeared in some Purkinje cells in pogo, but in only a few of Purkinje cells of their heterozygous littermate controls. In all groups of mice, no phospho-TH immunoreactive Purkinje cells were observed in the cerebellum, although subsets of TH immunoreactive Purkinje cells were found in adjacent sections. This study suggests that TH expression in the Purkinje cells of pogo abnormally increases without activation of this enzyme by phosphorylation. This may mean that TH in the Purkinje cells of these mutants does not catalyse the conversion of tyrosine to l -DOPA, and is not related to catecholamine synthesis. [source] Production of L -DOPA and dopamine in recombinant bacteria bearing the Vitreoscilla hemoglobin geneBIOTECHNOLOGY JOURNAL, Issue 7 2009Asli Giray Kurt Abstract Given the well-established beneficial effects of Vitreoscilla hemoglobin (VHb) on heterologous organisms, the potential of this protein for the production of L -DOPA and dopamine in two bacteria, Citrobacter freundii and Erwinia herbicola, was investigated. The constructed recombinants bearing the VHb gene (vgb+) had substantially higher levels of cytoplasmic L -DOPA (112 mg/L for C. freundii and 97 mg/L for E. herbicola) than their respective hosts (30.4 and 33.8 mg/L) and the vgb, control strains (35.6 and 35.8 mg/L). Further, the vgb+ recombinants of C. freundii and E. herbicola had 20-fold and about two orders of magnitude higher dopamine levels than their hosts, repectively. The activity of tyrosine phenol-lyase, the enzyme converting L -tyrosine to L -DOPA, was well-correlated to cytoplasmic L -DOPA levels. As cultures aged, higher tyrosine phenol-lyase activity of the vgb+ strains was more apparent. [source] Voltammetric Determination of L -Dopa on Poly(3,4-ethylenedioxythiophene)-Single-Walled Carbon Nanotube Composite Modified MicroelectrodesELECTROANALYSIS, Issue 4 2010Jayaraman Mathiyarasu Abstract In the present communication, it is shown that platinum microelectrodes electrochemically coated with a composite of poly(3,4-)ethylenedioxythiophene and single-walled carbon nanotubes (PEDOT/SWNT) enable determinations of 3,4-dihydroxy- L -phenylalaines (L -dopa) in neutral phosphate buffer solutions containing an excess of ascorbic acid. The interpenetrated networked nanostructure of the composite was characterized by scanning electron microscope (SEM) and Raman spectroscopy. It is shown that the presence of the composite gives rise to an increase in the electroactive area of an order of magnitude in compared to the area for the bare microelectrodes. The composite film-coated microelectrode, which yielded reversible cyclic voltammograms for the ferro/ferricyanide redox couple for scan rates between 0.01 and 0.10,V s,1, also gave rise to two well-resolved oxidation peaks for L -dopa and ascorbic acid (AA). The latter effect, which was not seen in the absence of the composite, enabled differential pulse voltammetric determinations of L -dopa in the concentration range between 0.1 to 20,,M with a detection limit of 100,nM. [source] Mutation of residues in the coenzyme binding pocket of Dopa decarboxylaseFEBS JOURNAL, Issue 10 2001Effects on catalytic properties Residues D271, H192, H302 and N300 of l -3,4-dihydroxyphenylalanine decarboxylase (DDC), a homodimeric pyridoxal 5,-phosphate (PLP) enzyme, were mutated in order to acquire information on the catalytic mechanism. These residues are potential participants in catalysis because they belong to the common PLP-binding structural motif of group I, II and III decarboxylases and other PLP enzymes, and because they are among the putative active-site residues of structural modelled rat liver DDC. The spectroscopic features of the D271E, H192Q, H302Q and N300A mutants as well as their dissociation constants for PLP suggest that substitution of each of these residues causes alteration of the state of the bound coenzyme molecule and of the conformation of aromatic amino acids, possibly in the vicinity of the active site. This supports, but does not prove, the possibility that these residues are located in the coenzyme-binding cleft. Interestingly, mutation of each residue generates an oxidative decarboxylase activity towards l -3,4-dihydroxyphenylalanine (l -Dopa), not inherent in the wild-type in aerobiosis, and reduces the nonoxidative decarboxylase activity of l -Dopa from 3- to 390-fold. The partition ratio between oxidative and nonoxidative decarboxylation ranges from 5.7 × 10,4 for N300A mutant to 946 × 10,4 for H302Q mutant. Unlike wild-type enzyme, the mutants catalyse these two reactions to the same extent either in the presence or absence of O2. In addition, all four mutants exhibit an extremely low level of the oxidative deaminase activity towards serotonin with respect to wild-type. All these findings demonstrate that although D271, H192, H302 and N300 are not essential for catalysis, mutation of these residues alters the nature of catalysis. A possible relationship among the integrity of the PLP cleft, the productive binding of O2 and the transition to a closed conformational state of DDC is discussed. [source] Influence of deep brain stimulation and levodopa on sensory signs in Parkinson's disease,MOVEMENT DISORDERS, Issue 9 2010Janne Gierthmühlen MD Abstract To examine the effects of levodopa (L -dopa) and deep brain stimulation of the subthalamic nucleus (STN-DBS) on sensory symptoms and signs in Parkinson's disease (PD). Seventeen patients with PD were included. (1) Presence of sensory symptoms and (2) effects of L -dopa and STN-DBS on sensory symptoms and signs [assessed by quantitative sensory testing (QST)] were examined 6 months after starting STN-DBS. In addition, in 12 of these patients, presence of sensory symptoms prior and post STN-DBS was compared. Pain was most frequently nociceptive. In about 30,40%, pain and sensory symptoms were associated with PD motor symptoms. In most of these cases, pain responded to L -dopa. Intensity of pain was reduced post STN-DBS compared to pre STN-DBS. L -Dopa had no influence on detection thresholds, whereas STN-DBS improved thermal detection thresholds. However, thermal and mechanical pain thresholds were uninfluenced by L -dopa or STN-DBS. Although some patients reported an improvement of pain with STN-DBS or L -dopa, objectively pain sensitivity as assessed by QST was not altered by STN-DBS or L -dopa suggesting that there is no evidence for a direct modulation of central pain processing by L -dopa or STN-DBS. © 2010 Movement Disorder Society [source] Ultrafast Energy Transfer from Bound Tetra(4- N,N,N,N -trimethylanilinium)porphyrin to Synthetic Dopa and Cysteinyldopa Melanins,PHOTOCHEMISTRY & PHOTOBIOLOGY, Issue 1 2003Tong Ye ABSTRACT The binding of tetra(4- N,N,N,N -trimethylanilinium)porphyrin (TAP) to melanins quenches the porphyrin emission. Time-resolved femtosecond absorption spectroscopy reveals that the mechanism behind this quenching is ultrafast nonradiative energy transfer (,ET < 100 fs) from electronically excited TAP to melanin. Similar dynamics are observed for both dopa and cysteinyldopa melanins. Steady-state emission studies demonstrate that the emission from melanin increases upon excitation of bound TAP, thereby confirming that rapid energy transfer occurs. These results are consistent with previous photoacoustic studies, which revealed that the TAP,melanin complex behaves like a supermolecular system liberating heat as a whole. [source] Use of neuroactive catecholamines to chemically induce metamorphosis of hatchery-reared flat oyster, Ostrea angasi, larvaeAQUACULTURE RESEARCH, Issue 14 2009Stephan O'Connor Abstract Low numbers and unreliable wild catch of the native flat oyster, Ostrea angasi, spat has resulted in the NSW flat oyster industry being reliant on hatchery-produced spat. The need to produce culchless spat in the hatchery stimulated investigation of several catecholamines to induce metamorphosis in O. angasi larvae. Larvae were treated with one of four neuroactive catecholamines (epinephrine, epinephrine bitartrate, l -Dopa and GABA) at one of four concentrations (10,3, 10,4, 10,5 or 10,6 m) for one of three treatment durations (0.5, 1,2 h) to determine morphogenic action for culchless spat production. Epinephrine bitartrate at 10,3 and 10,4 m and epinephrine at 10,3, 10,4 and 10,5 m, for a treatment duration of 1,2 h, produced significantly greater numbers of spat and culchless spat, compared with any other treatment combination. The other catecholamines tested did not induce a significant increase in the total number of spat or culchless spat, over untreated controls. Separate trials found that long-term treatment (24 h) with epinephrine bitartrate and epinephrine at morphogenic concentrations inhibited metamorphosis. Consecutive daily use of epinephrine bitartrate increased the numbers of spat and culchless spat produced, but did not affect larval or short-term post-larval survival. Treatment with 10,3 m epinephrine bitartrate or 10,4 m epinephrine for 1 h is recommended for routine commercial production of culchless flat oyster spat. [source] Voltammetric Determination of L -Dopa on Poly(3,4-ethylenedioxythiophene)-Single-Walled Carbon Nanotube Composite Modified MicroelectrodesELECTROANALYSIS, Issue 4 2010Jayaraman Mathiyarasu Abstract In the present communication, it is shown that platinum microelectrodes electrochemically coated with a composite of poly(3,4-)ethylenedioxythiophene and single-walled carbon nanotubes (PEDOT/SWNT) enable determinations of 3,4-dihydroxy- L -phenylalaines (L -dopa) in neutral phosphate buffer solutions containing an excess of ascorbic acid. The interpenetrated networked nanostructure of the composite was characterized by scanning electron microscope (SEM) and Raman spectroscopy. It is shown that the presence of the composite gives rise to an increase in the electroactive area of an order of magnitude in compared to the area for the bare microelectrodes. The composite film-coated microelectrode, which yielded reversible cyclic voltammograms for the ferro/ferricyanide redox couple for scan rates between 0.01 and 0.10,V s,1, also gave rise to two well-resolved oxidation peaks for L -dopa and ascorbic acid (AA). The latter effect, which was not seen in the absence of the composite, enabled differential pulse voltammetric determinations of L -dopa in the concentration range between 0.1 to 20,,M with a detection limit of 100,nM. [source] Electrochemical Behavior of Catecholamines and Related Compounds at In Situ Surfactant Modified Carbon Paste ElectrodesELECTROANALYSIS, Issue 2-3 2007M.Carmen Blanco-López Abstract The voltammetric characteristics of catecholamines: epinephrine (E) and norepinephrine (NE) and related compounds: isoproterenol, metanephrine, L -dopa, methyldopa, vanillylmandelic acid (VMA), and homovanillic acid (HVA) at unmodified and in situ surfactant- modified carbon paste electrodes were comparatively evaluated. For the basic and amphoteric compounds the modification of the electrode surface with submicellar concentrations of anionic surfactants (sodium dodecylsulfate, sodium decylsulfate or sodium dodecylsulfonate) produce an important current enhancement in its oxidation and reduction peak current together with the improvement in the reversibility of the processes. These effects were explained in basis on electrostatic and hydrophobic interactions. On the other hand, the oxidation of acidic metabolites, HVA and VMA, was studied at electrodes modified in situ with cationic surfactants. Under certain conditions the surfactant could stabilise some of the electrochemical reaction intermediates, thus explaining the different voltammetric behaviour of HVA and VMA. [source] Microwave Activation of Electrochemical Processes at Glassy Carbon and Boron-Doped Diamond ElectrodesELECTROANALYSIS, Issue 5-6 2005Kumar Sur, Ujjal Abstract Voltammetric experiments under intense microwave field conditions have been carried out at a carbon microfiber electrode, an array of carbon microfiber electrodes, and at a boron-doped diamond electrode. For the reversible one electron redox systems Fe(CN) and Ru(NH3) in aqueous KCl solution increased currents (up to 16 fold at a 33,,m diameter carbon microelectrode) and superheating (up to ca. 400,K at all types of electrodes) are observed. Electrodes with smaller diameter allow better signal enhancements to be achieved. From the missing effect of the supporting electrolyte concentration on the microwave enhanced currents, it can be concluded that effects observed at carbon electrodes (microwave absorbers) are due to the interaction of microwaves with the electrode material whereas for metal electrodes (microwave conductors) effects are dominated by the interaction of the microwaves with the aqueous dielectric. Short heat pulses can be applied by pulsing the microwave field and relatively fast temperature transients are observed for small electrodes. For the irreversible two electron oxidation of L -dopa in aqueous phosphate buffer, different types of effects are observed at glassy carbon and at boron-doped diamond. Arrays of carbon microfibers give the most reproducible and analytically useful current signal enhancements in the presence of microwaves. [source] Spectroelectrochemical and Voltammetric Studies of L -DOPAELECTROANALYSIS, Issue 2 2003Xiaoqiang Liu Abstract The electrooxidation of L -dopa at GC electrode was studied by in situ UV-vis spectroelectrochemistry (SEC) and cyclic voltammetry. The mechanism of electrooxidation and some reaction parameters were obtained. The results showed that the whole electrooxidation reaction of L -dopa at glassy carbon (GC) electrode was an irreversible electrochemical process followed by a chemical reaction in neutral solution (EC mechanism). The spectroelectrochemical data were treated by the double logarithm method together with nonlinear regression, from which the formal potential E0=228,mV, the apparent electron-transfer number of the electrooxidation reaction ,n=0.376 (R=0.99, SD=0.26), the standard electrochemical rate constant k0=(3.93±0.12)×10,4,cm s,1 (SD=1.02×10,2), and the formation equilibrium constant of the following chemical reaction kc=(5.38±0.34)×10,1,s,1 (SD=1.02×10,2) were also obtained. [source] Highly Enantioselective Synthesis of No-Carrier-Added 6-[18F]Fluoro- L -dopa by Chiral Phase-Transfer AlkylationEUROPEAN JOURNAL OF ORGANIC CHEMISTRY, Issue 13 2004Christian Lemaire Abstract [18F]Fluoro- L -dopa, an important radiopharmaceutical for positron emission tomography (PET), has been synthesized using a phase-transfer alkylation reaction. A chiral quaternary ammonium salt derived from a Cinchona alkaloid served as phase-transfer catalyst for the enantioselective alkylation of a glycine derivative. The active methylene group of this Schiff-base substrate was deprotonated with cesium hydroxide and rapidly alkylated by the 2-[18F]fluoro-4,5-dimethoxybenzyl halide (X = Br, I). The reaction proceeded with high yield (> 90%) at 0 °C or room temperature in various solvents such as toluene or dichloromethane. Preparation of the [18F]alkylating agent on a solid support was developed. After labelling, the labeled [18F]fluoroveratraldehyde was trapped on a tC18 cartridge and then converted on the cartridge into the corresponding benzyl halide derivatives by addition of aqueous sodium borohydride and gaseous hydrobromic or -iodic acid. Hydrolysis and purification by preparative HPLC made 6-[18F]fluoro- L -dopa ready for human injection in a 25,30% decay-corrected radiochemical yield in a synthesis time of 100 min. The product was found to be chemically, radiochemically and enantiomerically pure (ee > 95%). (© Wiley-VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2004) [source] The use of gene silencing to study the role of dopa decarboxylase in mosquito melanization reactionsINSECT MOLECULAR BIOLOGY, Issue 3 2005C.-Y. Huang Abstract Mosquito melanization involves hydroxylation of tyrosine to dopa, which then is oxidized to dopaquinone by phenoloxidase, or decarboxylated to dopamine by dopa decarboxlase (DDC). An Armigeres subalbatus cDNA encoding DDC was cloned and real-time PCR analysis revealed increased transcripts in blood-fed and microfilariae (mf)-inoculated mosquitoes. A double subgenomic Sindbis virus was used to silence DDC and assess its role in melanization of mf. DDC transcription and activity were significantly decreased in silenced mosquitoes, as was the degree of mf melanization 48 h postinoculation; however, melanization increased after 72 and 96 h, demonstrating that DDC influences the rate of melanization. DDC-silenced mosquitoes also exhibit high mortality, over-feeding and abnormal movement, consistent with an involvement of DDC in neurotransmission. [source] Aedes aegypti dopa decarboxylase: gene structure and regulationINSECT MOLECULAR BIOLOGY, Issue 3 2000M. T. Ferdig Abstract Dopa decarboxylase converts l -dopa to dopamine, a precursor molecule for diverse biological activities in insects including neurotransmission and a variety of tanning reactions required for development, reproduction and defence against parasites. Herein, we report the cloning and sequencing of the Aedes aegypti Ddc gene, including 2.1 kb of the upstream promoter region. The transcribed region of the gene spans more than 16 kb and contains five exons. In situ hybridization localizes the blood-meal-induced ovarian transcription of this gene to the follicular epithelial cells surrounding individual oocytes. Ovary tissue transcription of Ddc is increased in response to injection of 20-hydroxyecdysone to levels equal to those observed for blood-fed controls, however coinjection with the translational inhibitor cycloheximide negates the effect, indicating an indirect regulatory role for this hormone. Clusters of putative ecdysone-responsive elements and zinc-finger binding domains for the products of Broad-Complex gene family are identified in the 5,-promoter region. These elements are discussed in the context of common insect Ddc regulatory mechanisms. [source] | |||||||||||||||||||