Brain Dopamine Levels Research Articles

Juvenile hormone–dopamine systems for the promotion of flight activity in males of the large carpenter bee Xylocopa appendiculata

The reproductive roles of dopamine and dopamine regulation systems are known in social hymenopterans, but the knowledge on the regulation systems in solitary species is still needed. To test the possibility that juvenile hormone (JH) and brain dopamine interact to trigger territorial flight behavior in males of a solitary bee species, the effects on biogenic amines of JH analog treatments and behavioral assays with dopamine injections in males of the large carpenter bee Xylocopa appendiculata were quantified. Brain dopamine levels were significantly higher in methoprene-treated males than in control males 4 days after treatment, but were not significantly different after 7 days. Brain octopamine and serotonin levels did not differ between methoprene-treated and control males at 4 and 7 days after treatment. Injection of dopamine caused significantly higher locomotor activities and a shorter duration for flight initiation in experimental versus control males. These results suggest that brain dopamine can be regulated by JH and enhances flight activities in males. The JH-dopamine system in males of this solitary bee species is similar to that of males of the highly eusocial honeybee Apis mellifera.

Dissociations between interval timing and intertemporal choice following administration of fluoxetine, cocaine, or methamphetamine

The goal of our study was to characterize the relationship between intertemporal choice and interval timing, including determining how drugs that modulate brain serotonin and dopamine levels influence these two processes. In Experiment 1, rats were tested on a standard 40-s peak-interval procedure following administration of fluoxetine (3, 5, or 8mg/kg) or vehicle to assess basic effects on interval timing. In Experiment 2, rats were tested in a novel behavioral paradigm intended to simultaneously examine interval timing and impulsivity. Rats performed a variant of the bi-peak procedure using 10-s and 40-s target durations with an additional “defection” lever that provided the possibility of a small, immediate reward. Timing functions remained relatively intact, and ‘patience’ across subjects correlated with peak times, indicating a negative relationship between ‘patience’ and clock speed. We next examined the effects of fluoxetine (5mg/kg), cocaine (15mg/kg), or methamphetamine (1mg/kg) on task performance. Fluoxetine reduced impulsivity as measured by defection time without corresponding changes in clock speed. In contrast, cocaine and methamphetamine both increased impulsivity and clock speed. Thus, variations in timing may mediate intertemporal choice via dopaminergic inputs. However, a separate, serotonergic system can affect intertemporal choice without affecting interval timing directly.This article is part of a Special Issue entitled: Associative and Temporal Learning.

Reward-based hypertension control by a synthetic brain–dopamine interface

Synthetic biology has significantly advanced the design of synthetic trigger-controlled devices that can reprogram mammalian cells to interface with complex metabolic activities. In the brain, the neurotransmitter dopamine coordinates communication with target neurons via a set of dopamine receptors that control behavior associated with reward-driven learning. This dopamine transmission has recently been suggested to increase central sympathetic outflow, resulting in plasma dopamine levels that correlate with corresponding brain activities. By functionally rewiring the human dopamine receptor D1 (DRD1) via the second messenger cyclic adenosine monophosphate (cAMP) to synthetic promoters containing cAMP response element-binding protein 1(CREB1)-specific cAMP-responsive operator modules, we have designed a synthetic dopamine-sensitive transcription controller that reversibly fine-tunes specific target gene expression at physiologically relevant brain-derived plasma dopamine levels. Following implantation of circuit-transgenic human cell lines insulated by semipermeable immunoprotective microcontainers into mice, the designer device interfaced with dopamine-specific brain activities and produced a systemic expression response when the animal's reward system was stimulated by food, sexual arousal, or addictive drugs. Reward-triggered brain activities were able to remotely program peripheral therapeutic implants to produce sufficient amounts of the atrial natriuretic peptide, which reduced the blood pressure of hypertensive mice to the normal physiologic range. Seamless control of therapeutic transgenes by subconscious behavior may provide opportunities for treatment strategies of the future.

Pharmacokinetic and pharmacodynamic effects of methylphenidate and MDMA administered alone or in combination

Methylphenidate and 3,4-methylenedioxymethamphetamine (MDMA, 'ecstasy') are widely misused psychoactive drugs. Methylphenidate increases brain dopamine and norepinephrine levels by blocking the presynaptic reuptake transporters. MDMA releases serotonin, dopamine and norepinephrine through the same transporters. Pharmacodynamic interactions of methylphenidate and MDMA are likely. This study compared the pharmacodynamic and pharmacokinetic effects of methylphenidate and MDMA administered alone or in combination in healthy subjects using a double-blind, placebo-controlled, crossover design. Methylphenidate did not enhance the psychotropic effects of MDMA, although it produced psychostimulant effects on its own. The haemodynamic and adverse effects of co-administration of methylphenidate and MDMA were significantly higher compared with MDMA or methylphenidate alone. Methylphenidate did not change the pharmacokinetics of MDMA and vice versa. Methylphenidate and MDMA shared some subjective amphetamine-type effects; however, 125 mg of MDMA increased positive mood more than 60 mg of methylphenidate, and methylphenidate enhanced activity and concentration more than MDMA. Methylphenidate and MDMA differentially altered facial emotion recognition. Methylphenidate enhanced the recognition of sad and fearful faces, whereas MDMA reduced the recognition of negative emotions. Additionally, the present study found acute pharmacodynamic tolerance to MDMA but not methylphenidate. In conclusion, the combined use of methylphenidate and MDMA does not produce more psychoactive effects compared with either drug alone, but potentially enhances cardiovascular and adverse effects. The findings may be of clinical importance for assessing the risks of combined psychostimulant misuse. Trial registration identification number: NCT01465685 (http://clinicaltrials.gov/ct2/show/NCT01465685).

Elevation of Dopamine Induced by Cigarette Smoking: Novel Insights from a [11C]-(+)-PHNO PET Study in Humans

Positron emission tomography (PET) has convincingly provided in vivo evidence that psychoactive drugs increase dopamine (DA) levels in human brain, a feature thought critical to their reinforcing properties. Some controversy still exists concerning the role of DA in reinforcing smoking behavior and no study has explored whether smoking increases DA concentrations at the D3 receptor, speculated to have a role in nicotine's addictive potential. Here, we used PET and [(11)C]-(+)-PHNO ([(11)C]-(+)-4-propyl-3,4,4a,5,6,10b-hexahydro-2H-naphtho[1,2-b][1,4]oxazin-9-ol) to test the hypothesis that smoking increases DA release (decreases [(11)C]-(+)-PHNO binding) in D2-rich striatum and D3-rich extra-striatal regions and is related to craving, withdrawal and smoking behavior. Ten participants underwent [(11)C]-(+)-PHNO scans after overnight abstinence and after smoking a cigarette. Motivation to smoke (smoking topography), mood, and craving were recorded. Smoking significantly decreased self-reported craving, withdrawal, and [(11)C]-(+)-PHNO binding in D2 and D3-rich areas (-12.0 and -15.3%, respectively). We found that motivation to smoke (puff rate) predicted magnitude of DA release in limbic striatum, and the latter was correlated with decreased craving and withdrawal symptoms. This is the first report suggesting that, in humans, DA release is increased in D3-rich areas in response to smoking. Results also support the preferential involvement of the limbic striatum in motivation to smoke, anticipation of pleasure from cigarettes and relief of withdrawal symptoms. We propose that due to the robust effect of smoking on [(11)C]-(+)-PHNO binding, this radiotracer represents an ideal translational tool to investigate novel therapeutic strategies targeting DA transmission.

Flattening plasma corticosterone levels increases the prevalence of serotonergic dorsal raphe neurons inhibitory responses to nicotine in adrenalectomised rats

Major depression is characterized by a diminished activity of the brain serotonergic system as well as by the flattening of plasma cortisol levels. Nicotine improves mood in patients with major depression and in experimentally depressed animals by increasing brain serotonin (5-HT), noradrenaline and dopamine levels. The present study was directed to determine if flattening plasma glucocorticoid levels changes nicotine's stimulatory effects upon 5-HT DRN neurons. The experiments were performed in brain slices obtained from rats previously (14 days) adrenalectomised and implanted subcutaneously with one pellet containing 75mg of corticosterone (Adx+CSR rats). Whole cell voltage and current clamp techniques were used to study the activity of immunocitochemically identified 5-HT DRN neurons. Administration of nicotine (1μM) in sham-operated animals produced stimulatory effects in all 5-HT DRN neurons studied. In Adx+CSR rats however, nicotine inhibited 75% of 5-HT DRN neurons and increased the potassium-dependent inward rectifying current. The inhibitory effect of nicotine upon 5-HT DRN neurons was dependent on serotonin release inside the DRN, since it was converted into a stimulatory response by the selective antagonist of 5-HT1A receptors N-[2-[4-(2-methoxyphenyl)-1-piperazinyl]ethyl]-N-(2-pyridyl)cyclohexanecarboxamide (WAY100635, 25nM). Adx+CSR rats also presented an increased function of 5-HT1A autoreceptors, since, in these rats, serotonin (1-10μM) produced a higher increase in the potassium dependent inward rectifying current in comparison with sham-operated animals. Serotonin release inside DRN was mediated by α4β2 nicotinic acetylcholine receptors since the selective antagonist of these receptors dihydro-β-erytroidine hydrobromide (DHβE, 100nM) blocked the inhibitory effects of nicotine 5-HT DRN neurons. These data indicate that, in the experimental model of adrenalectomised rats implanted with corticosterone pellets, nicotine increases the function of 5-HT1A receptors of 5-HT DRN neurons.

Systemic and brain bioavailabilities of d-Phenylglycine-l-Dopa, a sustained dopamine-releasing prodrug

d-Phenylglycine-l-Dopa (d-PhG-l-Dopa), designed as a dipeptide mimetic prodrug of dopamine, has been proven to have 31-fold higher oral bioavailability than l-Dopa in rats. To further investigate if this dipeptide enters the brain, a single-dose pharmacokinetic study by i.v. administration, in comparison with l-Dopa, was conducted to monitor brain availability. The dopamine level in the brain was also determined. The results indicated that both d-PhG-l-Dopa and l-Dopa entered the brain rapidly (Tmax 1 minute) with similar degrees of penetration (AUCbrain/AUCplasma 8.83% vs. 7.61%). As d-PhG-l-Dopa had higher systemic exposure than l-Dopa (AUCplasma 23.79 μmol*min/mL vs. 12.09 μmol*min/mL), it thus had higher brain availability (AUCbrain 2.0 μmol*min/mL vs. 0.92 μmol*min/mL). Although the AUCbrain dopamine after d-PhG-l-Dopa treatment was only 24% of that from l-Dopa treatment, it however exhibited higher anti-Parkinsonism activity than l-Dopa (reduction in rotation number 47.9 ± 5.5% vs. 27.3 ± 4.8%). Higher brain dopamine residual properties of d-PhG-l-Dopa treatment compared to l-Dopa treatment, namely 3.2 times longer MRTbrain dopamine (172.15 vs. 53.78 minutes), 10 times longer Tmax brain dopamine (30 vs. 3 minutes) and 8.36 times longer half-life (T1/2Tmaxtoend(min) 112.51 vs. 13.46 minutes), may explain the result. Moreover, the long terminal half-life of brain dopamine upon d-PhG-l-Dopa treatment indicated its slow dopamine-releasing property compared with l-Dopa treatment (112.51 vs. 44.85 minutes). It is also important to note that brain dopamine level was better controlled in the d-PhG-l-Dopa group than in the l-Dopa group (Cmax/Cmin 1.62 vs. 9.85). In conclusion, this study demonstrated that d-PhG-l-Dopa is an intrinsic dopamine-sustained-releasing prodrug. The limited concentration fluctuation of released dopamine in the brain is beneficial for the clinical management of Parkinson's disease.

5,7-Dihydroxy-6-methoxy-4'-phenoxyflavone, a derivative of oroxylin A improves attention-deficit/hyperactivity disorder (ADHD)-like behaviors in spontaneously hypertensive rats

Oroxylin A, a major flavonoid in Scutellaria baicalensis, has been shown to alleviate attention-deficit/hyperactivity disorder (ADHD)-like behaviors in the spontaneously hypertensive rat (SHR) model of ADHD. As part of our continuing effort to discover effective ADHD drug candidates, we synthesized a number of oroxylin A derivatives and characterized their biological activities. Among all oroxylin A analogs, compound 7-7 (5,7-dihydroxy-6-methoxy-4'-phenoxyflavone) showed the most remarkable inhibition of dopamine reuptake alike methylphenidate, a dopamine transporter (DAT) blocker and typical drug for ADHD, and oroxylin A. It did not influence norepinephrine reuptake unlike atomoxetine, a selective norepinephrine inhibitor. Moreover, compound 7-7 reduced hyperactivity, sustained inattention and impulsivity in the SHR as measured by the open field, Y-maze and electro-foot shock aversive water drinking tests, respectively. Most drugs that enhance brain dopamine levels (e.g. DAT blockers like cocaine and methylphenidate) produce behavioral effects like those of stimulants causing them to be abused. However, the repeated treatment of compound 7-7 failed to elicit locomotor sensitization in rats, and neither produced conditioned place preference response nor maintained self-administration behavior. Altogether, the present study suggests the promising therapeutic value of compound 7-7 as an ADHD drug. Furthermore, compound 7-7 may be considered as an alternative therapy to psychostimulant ADHD treatments (e.g. amphetamine and methylphenidate) for which use has been deemed controversial due to their abuse liability.

Endocrine Disruption in the European Eel, Anguilla anguilla, Exposed to an Environmental Cocaine Concentration

The aim of the present study was to verify if cocaine, at environmental concentrations, influences the endocrine system of the European eel. Silver eels (a stage of the eel life cycle preparing the fish for the oceanic reproductive migration) were exposed to a nominal cocaine concentration of 20 ng/l during 30 days; at the same time, control, carrier, and postexposure recovery groups were made. The effects of cocaine were observed in (1) brain dopamine content, (2) plasma catecholamine levels (dopamine, norepinephrine, and epinephrine), (3) pituitary–adrenal axis activity [plasma adrenocorticotropic hormone (ACTH), corticosterone, cortisol, and aldosterone levels], and (4) pituitary–thyroid axis activity [plasma thyroid-stimulating hormone (TSH), triiodothyronine, and thyroxine levels]. In the treated group, brain dopamine, plasma catecholamines, cortisol, and TSH levels were higher, whereas ACTH, corticosterone, and triiodothyronine levels were lower than controls. In the postexposure recovery group, brain dopamine, plasma dopamine and epinephrine, and thyroxine levels further increased, whereas plasma norepinephrine, cortisol, and corticosterone levels were similar to treated values. Finally, ACTH and TSH were similar, whereas triiodothyronine levels were lower than controls. Aldosterone levels were unaffected by cocaine exposure. The results of the present study show that cocaine, at environmental concentrations, behaves like an endocrine disruptor changing brain dopamine and plasma catecholamine levels and the activity of pituitary–adrenal/thyroid axes. Since the endocrine system plays a key role in the metabolic and reproductive processes of the eel, our results suggest that environmental cocaine could be considered another cause for the decline in the European eel.

Amelioration of Behavioral Abnormalities in BH4-deficient Mice by Dietary Supplementation of Tyrosine

This study reports an amelioration of abnormal motor behaviors in tetrahydrobiopterin (BH4)-deficient Spr −/− mice by the dietary supplementation of tyrosine. Since BH4 is an essential cofactor for the conversion of phenylalanine into tyrosine as well as the synthesis of dopamine neurotransmitter within the central nervous system, the levels of tyrosine and dopamine were severely reduced in brains of BH4-deficient Spr −/− mice. We found that Spr −/− mice display variable ‘open-field’ behaviors, impaired motor functions on the ‘rotating rod’, and dystonic ‘hind-limb clasping’. In this study, we report that these aberrant motor deficits displayed by Spr −/− mice were ameliorated by the therapeutic tyrosine diet for 10 days. This study also suggests that dopamine deficiency in brains of Spr −/− mice may not be the biological feature of aberrant motor behaviors associated with BH4 deficiency. Brain levels of dopamine (DA) and its metabolites in Spr −/− mice were not substantially increased by the dietary tyrosine therapy. However, we found that mTORC1 activity severely suppressed in brains of Spr −/− mice fed a normal diet was restored 10 days after feeding the mice the tyrosine diet. The present study proposes that brain mTORC1 signaling pathway is one of the potential targets in understanding abnormal motor behaviors associated with BH4-deficiency.

Dopaminergic effect of Alstonia scholaris Linn. R.Br. in experimentally induced anxiety

Aim of the studyThe anti-anxiety activity of the ethanolic extract of leaves of Alstonia scholaris (Family: Apocynaceae) has been well established. The aim of the present study is to evaluate the anti-anxiety activity of the dichloromethane fraction (known as DCM) fractionated from ethanolic extract of leaves of A. scholaris (known as EEAS). Materials and methodsDCM fraction (10 and 30mg/kg) was tested against various anti-anxiety models viz. elevated plus maze, open field, hole board, light dark test, mirror chamber and foot shock induced aggression models. The false positive results were overruled by actophotometer and rotarod tests. The change in brain monoamines was estimated. The possible dopaminergic effect was tested by haloperidol-induced catalepsy and 6-hydroxy dopamine induced Parkinsonism. ResultsThe DCM fraction of EEAS was found to be significantly active in all the tested models of anxiety, which confirmed their anti-anxiety activity. Lack of significant change in locomotion test indicated that DCM did not have stimulant or sedative effects. There was also no significant change in the time spent in the rotarod, which indicated its lack of motor or muscle in coordination. Increase in brain dopamine levels and reversal of dopamine-induced catalepsy confirmed that the mechanism of anti-anxiety activity is mediated through dopamine. The effect of DCM in 6-hydroxy dopamine-induced anxiety was conclusive of its dopaminergic activity. ConclusionThe present study suggests that DCM possess anti-anxiety and the apparent mechanism of action is through dopaminergic mediation.

Neuroendocrine mechanisms underlying regulation of mating flight behaviors in male honey bees (Apis mellifera L.)

We determined the neuroendocrine mechanisms underlying regulation of mating flight behaviors in male honey bees. Both a precursor of dopamine (3,4-dihydroxyphenylalanine: DOPA) and a precursor of octopamine (tyramine) in the brain decreased in an age-dependent fashion before sexual maturation (i.e. 8days of age), whereas the levels of brain dopamine, dopamine metabolites (N-acetyldopamine and norepinephrine) and octopamine were increased. These age-dependent increases of dopamine and octopamine were also detected in the meso-metathoracic ganglia. Injection of either dopamine or octopamine into 7–8-day-old males shortened the duration for flight-initiation and increased the duration of wing vibration, indicating that both dopamine and octopamine enhance the flight-initiation and -sustaining activities in males. Applications of a juvenile hormone analog (methoprene) enhanced the levels of dopamine in the brains of 4-day-old males, but this enhancement was not detected in either brain octopamine or meso-metathoracic dopamine and octopamine. Thus, we found that both dopamine and octopamine in the brain and meso-metathoracic ganglia increase until sexual maturation and could enhance the activities of mating flight independently; in addition, the increase in levels of dopamine in the brain could be selectively regulated by juvenile hormone. The regulatory systems of dopamine and octopamine in honey bee males might be ‘classical’ and similar to those of primitively eusocial hymenopterans, and partly adapt to the short lifespan with a single mating system in the males.

COMTrs4680 Met is not always the ‘smart allele’: Val allele is associated with better working memory and larger hippocampal volume in healthy Chinese

Catechol-O-methyltransferase (COMT) Val158Met (rs4680) polymorphism plays a crucial role in regulating brain dopamine level. Converging evidence from Caucasian samples showed that, compared with rs4680 Val allele, the Met allele was linked to lower COMT activity, which in turn was linked to better cognitive performance such as working memory (WM) and to a larger hippocampus (a brain region important for WM). However, some behavioral studies have shown that the function of rs4680 appears to vary across different ethnic groups, with Chinese subjects showing an opposite pattern as that for Caucasians (i.e. the Val allele is linked to better cognitive functions related to WM in Chinese). Using a sample of healthy Han Chinese college students (ages from 19 to 21 years), this study investigated the association of COMT Val158Met genotype with behavioral data on a two-back WM task (n = 443, 189M/254F) and T1 MRI data (n = 320, 134M/186F). Results showed that, compared to the Met allele, the Val allele was associated with larger hippocampal volume (the right hippocampus: β = -0.118, t = -2.367, P = 0.019, and the left hippocampus: β = -0.099, t = -1.949, P = 0.052) and better WM performance (β = -0.110, t = -2.315, P = 0.021). These results add to the growing literature on differentiated effects of COMT rs4680 polymorphism on WM across populations and offer a brain structural mechanism for such population-specific genetic effects.

Brood-care experience, nursing performance, and neural development in the ant Pheidole dentata

Social insect workers mature behaviorally and physiologically with increasing age, generally transitioning from or adding new tasks to their existing repertoire of within-nest nursing tasks. As adult minor workers of the ant Pheidole dentata age, they attend to brood more frequently and nurse more efficiently, perform a broader array of tasks, and undergo myological and neural development. Because these factors covary, the causal relationships among age, task experience, and neural and physiological maturation are not understood. We compared brood-care performance and efficiency by 10-day-old P. dentata minors that had acquired nursing experience to that of equal-age minors experimentally deprived of brood contact. We found the frequency and efficiency of nursing did not significantly differ between experimental and control worker groups, suggesting experience is not required for age-related improvement in nursing efficiency. Workers with and without prior nursing experience did not significantly differ macroscopically in brain anatomy or in brain serotonin content, although workers from the two treatments had slightly, but significantly, different levels of brain dopamine. These results suggest experience with brood is not required for P. dentata minor workers to develop nursing proficiency or undergo a substantial degree of the age-related neural development identifiable by our assessments, which could underscore the ontogeny of brood-care efficiency.

Presynaptic regulation of extracellular dopamine levels in the medial prefrontal cortex and striatum during tyrosine depletion

Available neurochemical probes that lower brain dopamine (DA) levels in man are limited by their tolerability and efficacy. For instance, the acute lowering of brain tyrosine is well tolerated, but only modestly lowers brain DA levels. Modification of tyrosine depletion to robustly lower DA levels would provide a superior research probe. The objective of this study was to determine whether the subthreshold stimulation of presynaptic DA receptors would potentiate tyrosine depletion-induced effects on extracellular DA levels in the medial prefrontal cortex (MPFC) and striatum of the rat. We administered quinpirole, a predominantly DA type 2 (D2R) receptor agonist, into the MPFC and striatum by reverse dialysis. A tyrosine- and phenylalanine-free neutral amino acid mixture [NAA(-)] IP was used to lower brain tyrosine levels. DA levels in the microdialysate were measured by HPLC with electrochemical detection. Quinpirole dose-dependently lowered DA levels in MPFC as well as in the striatum. NAA(-) alone transiently lowered DA levels (80% baseline) in the striatum, but had no effect in MPFC. The co-administration of NAA(-) and a subthreshold concentration of quinpirole (6.25nM) lowered DA levels (50% baseline) in both the MPFC and striatum. This effect was blocked by the mixed D2R/D3R antagonist haloperidol at IP doses that on their own did not affect DA levels (10.0nmol/kg in the MPFC and 0.10nmol/kg in the striatum). Pharmacological stimulation of inhibitory D2R receptors during tyrosine depletion markedly lowers the extracellular DA levels in the MPFC and striatum. The data suggest that combining tyrosine depletion with a low dose of a DA agonist should robustly lower brain regional DA levels in man.

The effects of L-Dopa and <i>N</i>-(alpha-linolenoyl) tyrosine on 6-OH-DA lesions on dopamine level and activity, dyskinesia and homocysteinemia in rats

The three major issues in L-DOPA therapy for Parkinson are: 1) rapid tolerance; 2) induced dyskinesia; 3) hyperhomocysteinemia. The newly synthesized molecule, which is an amide bond molecule composed of L-tyrosine and alphalinolenic acid, i.e., N-(alpha-linolenoyl) tyrosine (NLT), represents a new class of molecules that combines essential amino acids with essential fatty acids. This study demonstrates that acute administration of NLT results in prolonged increase of the brain dopamine level, and that this molecule is able to overcome the three major problems associated with Parkinson’s L-DOPA therapy: it is able to elevate brain dopamine levels and increase activity, without inducing tolerance, dyskinesia or hyperhomocysteinemia.

The Effect of Chronic Tobacco Smoking and Chewing on the Lipid Profile

A worldwide epidemic of cardio vascular diseases is evolving, out of which atherosclerosis appears to be the most frequent underlying cause. Cigarette smoking remains the most important cause of the preventable morbidity and the early mortality. Nicotine is highly addictive, it raises the brain levels of dopamine and it produces withdrawal symptoms on its discontinuation. To study the effect of tobacco smoking & chewing on serum lipid profile. Although a genetic predis-position to atherosclerosis may be the cause, a vast majority of the atherosclerotic related diseases, which include coronary heart diseases, are acquired. Those which usually appear later in life are largely preventable. Tobacco is the major and the single most preventable risk factor for atherosclerotic related, clinical events like coronary heart disease. This study was conducted on three groups of male subjects, with each group containing 25 individuals of 25 to 35 years of age and who weighed 50-70 kgs.Group-I: non smokers and non chewers.Group-II: smokers and non chewersGroup-III: chewers and non smokers.To estimate the triglycerides, glycerol which is derived from the saponification of triglycerides is oxidized to formaldehyde, which in turn is made to react with ammonia and acetylacetone to give rise to a chromogen (3.5 diacetyle-1,4 dihydrolutidine). It is quantified spectro-photometrically (the HANTZSCH reaction). The mean serum total cholesterol level in the subjects of Group II was more by about 16.94 % (p< 0.001) and that in the subjects of Group -III was more by 23.21% (p< 0.001).The mean serum VLDL level in the subjects of Group II had an increase of about 27.54% (p< 0.01) and in Group -III, it had increased by11.82% (p< 0.01).The mean serum LDL level in the subjects of Group II showed an increase of about 34.64% (p< 0.001) and in Group -III, it had increased by16.27% (p< 0.001).The mean serum HDL level in the subjects of Group II showed a decrease in the mean serum HDL level by about 9.78 % (p< 0.01) and in Group -III, it had decreased by 22.12% (p< 0.01).The mean serum Triglyceride level in the subjects of Group II showed an increase of about 25.40% (p< 0.001) and in Group -III, it was more by33.35% (p< 0.001). There was a significant increase in total cholesterol and LDL-C in tobacco users ,as compared to non tobacco users.

Regulation of the dopaminergic system in a murine model of aromatic l-amino acid decarboxylase deficiency

Aromatic l-amino acid decarboxylase (AADC) is responsible for the syntheses of dopamine and serotonin. Children with AADC deficiency exhibit compromised development, particularly with regard to their motor functions. Currently, no animal model of AADC deficiency exists. We inserted an AADC gene mutation (IVS6+4A>T) and a neomycin-resistance gene into intron 6 of the mouse AADC (Ddc) gene. In the brains of homozygous knock-in (KI) mice (DdcIVS6/IVS6), AADC mRNA lacked exon 6, and AADC activity was <0.3% of that in wild-type mice. Half of the KI mice were born alive but grew poorly and exhibited severe dyskinesia and hindlimb clasping after birth. Two-thirds of the live-born KI mice survived the weaning period, with subsequent improvements in their growth and motor functions; however, these mice still displayed cardiovascular dysfunction and behavioral problems due to serotonin deficiencies. The brain dopamine levels in the KI mice increased from 9.39% of the levels in wild-type mice at 2weeks of age to 37.86% of the levels in wild-type mice at 8weeks of age. Adult KI mice also exhibited an exaggerated response to apomorphine and an elevation of striatal c-Fos expression, suggesting post-synaptic adaptations. Therefore, we generated an AADC deficient mouse model, in which compensatory regulation allowed the mice to survive to adulthood. This mouse model will be useful both for developing gene therapies for AADC deficiency and for designing treatments for diseases associated with neurotransmitter deficiency.

Additive inhibition of human α1β2γ2 GABAA receptors by mixtures of commonly used drugs of abuse

Yearly, exposure to drugs of abuse results in ∼1 million emergency department visits in the US. In ∼50% of the visits, stimulant drugs like cocaine and amphetamine-like substances (e.g. 3,4-methylenedioxymethamphetamine (MDMA, the main active ingredient of ecstasy)) are involved, whereas in ∼60% multiple drugs are involved. These drugs induce higher dopamine and serotonin levels resulting in drug-induced toxicity. Since GABA receptors (GABA-R) provide the main inhibitory input on dopaminergic and serotonergic neurons, drug-induced inhibition of GABA-R could contribute to higher neurotransmitter levels and thus toxicity.We therefore investigated the effects of combinations of commonly abused stimulant drugs (cocaine, MDMA, 3,4-methylenedioxyamphetamine (MDA) and meta-chlorophenylpiperazine (mCPP)) on the function of the human α1β2γ2 GABAA receptor (hGABAA-R), expressed in Xenopus oocytes, using the two-electrode voltage-clamp technique.These drugs concentration-dependently inhibited the GABA-evoked current (mCPP>cocaine>MDMA>MDA). Most drug combinations decreased the GABA-evoked current stronger than the single drug. Additivity was observed during combined exposure to low concentrations of cocaine and mCPP as well as during combined exposure to MDA with cocaine or mCPP. However, combinations containing MDMA mainly resulted in sub-additivity or no additivity.At drug concentrations relevant for clinical toxicology, co-exposure to ≥2 drugs can decrease the GABA-evoked current in an additive manner. Thus, in patients exposed to multiple drugs, inhibitory GABA-ergic input is reduced more prominently, likely resulting in higher brain dopamine levels. As this will increase the risk for drug-induced toxicity, treatment of drug-intoxicated patients with drugs that enhance GABA-ergic input should be further optimized.

Interactions of anti-Parkinson drug benserazide with Zn(II), Cu(II), Fe(II) ions

One of the treatments of Parkinson disease is based on increasing the brain dopamine level by L-DOPA (LD) applications. To prevent the peripheral degradation of levodopa, another drug, benserazide is applied. On the other hand, during this neurodegenerative disease changes in the homeostasis of metals are observed and the increasing brain zinc levels are postulated to have therapeutic effects. Here we present studies on interactions of Zn(II), Cu(II), Fe(II) ions with benserazide and with benserazide/levodopa in ternary system. By applying mass spectrometry and UV–vis methods we describe the interactions between selected metal ions and the drug additives in the investigated systems. The results show forming of equimolar complexes in the binary and ternary systems.