Unabated occupational risk in a patient with rheumatoid pulmonary fibrosis.

BACKGROUND: This case highlights the importance of considering hypersensitivity pneumonitis (HP) in the differential diagnosis of interstitial lung disease (ILD) and of obtaining an occupational history so that remediable risk factors may be identified and managed. AIMS: To report a case of a chicken sexer with severe rheumatoid arthritis (RA) who developed progressively worsening dyspnoea and restrictive lung disease associated with pulmonary fibrosis. METHODS: Clinical investigation included physical examination, occupational history, pulmonary function tests (PFTs), chest imaging and bronchoalveolar lavage (BAL), as well as serological tests including standard IgE bird feather mixture and local IgG precipitin preparation to chicken excrement. Lung histopathology was examined post-mortem. RESULTS: The patient had worked as a chicken sexer for 29 years with limited control of exposure to chicken bioaerosols. PFTs initially showed mild restriction with a moderate gas transfer defect and computerized tomography of the chest exhibited extensive interstitial infiltrates throughout with severe honeycombing at the bases. Cytology from a BAL revealed multinucleated giant cells (MNGs). Specific serologic tests for bird antigens were negative. Histopathology demonstrated diffuse interstitial fibrosis with honeycombing, poorly formed granulomas and MNGs. CONCLUSIONS: Findings were consistent with a diagnosis of HP with RA-associated ILD. The patient's history of severe RA biased the diagnosis to one of RA-associated ILD and her occupational risk had been less emphatically addressed. Obtaining a thorough occupational history can uncover exposures to workplace respiratory hazards and may create opportunities for intervention to limit morbidity from chronic lung disease.

Role of oxidative changes in the degeneration of dopamine terminals after injection of neurotoxic levels of dopamine.

Dopamine may contribute to the loss of dopamine neurons in Parkinson's disease by generating reactive oxygen species and quinones. A previous report from this laboratory showed that intrastriatal injection of dopamine resulted in the selective reduction of tyrosine hydroxylase immunoreactivity, accompanied by an increase in indices of dopamine oxidation. However, conclusive proof that decreased tyrosine hydroxylase immunoreactivity represented a loss of dopamine terminals was lacking. In this paper, we demonstrate that injection of dopamine results in a selective loss of dopamine terminals by (i) showing that immunoreactivity for another selective marker for dopamine terminals, the dopamine transporter, is also reduced; and (ii) that amino-cupric-silver stain reveals terminal degeneration within the area of selective loss of dopamine terminals. To determine the dopamine concentration that is selectively toxic to dopamine terminals, we examined changes in extracellular dopamine and 3,4-dihydroxyphenylacetic acid in the area of selective terminal loss following intrastriatal dopamine. Dopamine and 3,4-dihydroxyphenylacetic acid in this region reached peak levels 1-2h after the injection, and then returned towards baseline. The peak level of dopamine in the area of selective dopamine terminal damage was 10(2)-10(3)-fold lower than the injected concentration. Changes in striatal tissue levels of cysteinyl-catechols and glutathione were examined at 2, 4, 8, and 24h after intrastriatal dopamine. Levels of protein cysteinyl-dopamine and cysteinyl-3,4-dihydroxyphenylacetic acid were increased at all time-points following the dopamine injection. High levels of free cysteinyl-catechols and glutathione-dopamine were detected within 2h after the dopamine injection. Glutathione levels were decreased significantly at 4 and 8h after the injection of dopamine, and returned to control levels by 24h. These data indicate that dopamine terminals actively degenerate following a single intrastriatal injection of dopamine, and furthermore that oxidative stress plays a key role in this process. As oxidative stress is thought to play an active role in the pathobiology of Parkinson's disease, these data may be relevant to our understanding of the disorder.

Quantitative biochemical and ultrastructural comparison of mitochondrial permeability transition in isolated brain and liver mitochondria: evidence for reduced sensitivity of brain mitochondria.

Opening of the mitochondrial permeability transition pore has increasingly been implicated in excitotoxic, ischemic, and apoptotic cell death, as well as in several neurodegenerative disease processes. However, much of the work directly characterizing properties of the transition pore has been performed in isolated liver mitochondria. Because of suggestions of tissue-specific differences in pore properties, we directly compared isolated brain mitochondria with liver mitochondria and used three quantitative biochemical and ultrastructural measurements of permeability transition. We provide evidence that brain mitochondria do not readily undergo permeability transition upon exposure to conditions that rapidly induce the opening of the transition pore in liver mitochondria. Exposure of liver mitochondria to transition-inducing agents led to a large, cyclosporin A-inhibitable decrease in spectrophotometric absorbance, a loss of mitochondrial glutathione, and morphologic evidence of matrix swelling and disruption, as expected. However, we found that similarly treated brain mitochondria showed very little absorbance change and no loss of glutathione. The absence of response in brain was not simply due to structural limitations, since large-amplitude swelling and release of glutathione occurred when membrane pores unrelated to the transition pore were formed. Additionally, electron microscopy revealed that the majority of brain mitochondria appeared morphologically unchanged following treatment to induce permeability transition. These findings show that isolated brain mitochondria are more resistant to induction of permeability transition than mitochondria from liver, which may have important implications for the study of the mechanisms involved in neuronal cell death.

Peroxynitrite- and nitrite-induced oxidation of dopamine: implications for nitric oxide in dopaminergic cell loss.

Increased nitric oxide (NO) production has been implicated in many examples of neuronal injury such as the selective neurotoxicity of methamphetamine and 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine to dopaminergic cells, presumably through the generation of the potent oxidant peroxynitrite (ONOO). Dopamine (DA) is a reactive molecule that, when oxidized to DA quinone, can bind to and inactivate proteins through the sulfhydryl group of the amino acid cysteine. In this study, we sought to determine if ONOO could oxidize DA and participate in this process of protein modification. We measured the oxidation of the catecholamine by following the binding of [3H]DA to the sulfhydryl-rich protein alcohol dehydrogenase. Results showed that ONOO oxidized DA in a concentration- and pH-dependent manner. We confirmed that the resulting DA-protein conjugates were predominantly 5-cysteinyl-DA residues. In addition, it was observed that ONOO decomposition products such as nitrite were also effective at oxidizing DA. These data suggest that the generation of NO and subsequent formation of ONOO or nitrite may contribute to the selective vulnerability of dopaminergic neurons through the oxidation of DA and modification of protein.

Dopamine oxidation alters mitochondrial respiration and induces permeability transition in brain mitochondria: implications for Parkinson's disease.

Both reactive dopamine metabolites and mitochondrial dysfunction have been implicated in the neurodegeneration of Parkinson's disease. Dopamine metabolites, dopamine quinone and reactive oxygen species, can directly alter protein function by oxidative modifications, and several mitochondrial proteins may be targets of this oxidative damage. In this study, we examined, using isolated brain mitochondria, whether dopamine oxidation products alter mitochondrial function. We found that exposure to dopamine quinone caused a large increase in mitochondrial resting state 4 respiration. This effect was prevented by GSH but not superoxide dismutase and catalase. In contrast, exposure to dopamine and monoamine oxidase-generated hydrogen peroxide resulted in a decrease in active state 3 respiration. This inhibition was prevented by both pargyline and catalase. We also examined the effects of dopamine oxidation products on the opening of the mitochondrial permeability transition pore, which has been implicated in neuronal cell death. Dopamine oxidation to dopamine quinone caused a significant increase in swelling of brain and liver mitochondria. This was inhibited by both the pore inhibitor cyclosporin A and GSH, suggesting that swelling was due to pore opening and related to dopamine quinone formation. In contrast, dopamine and endogenous monoamine oxidase had no effect on mitochondrial swelling. These findings suggest that mitochondrial dysfunction induced by products of dopamine oxidation may be involved in neurodegenerative conditions such as Parkinson's disease and methamphetamine-induced neurotoxicity.

Cytotoxic and genotoxic potential of dopamine.

A variety of in vitro and in vivo studies demonstrate that dopamine is a toxic molecule that may contribute to neurodegenerative disorders such as Parkinson's disease and ischemia-induced striatal damage. While much attention has focused on the fact that the metabolism of dopamine produces reactive oxygen species (peroxide, superoxide, and hydroxyl radical), growing evidence suggests that the neurotransmitter itself may play a direct role in the neurodegenerative process. Oxidation of the dopamine molecule produces a reactive quinone moiety that is capable of covalently modifying and damaging cellular macromolecules. This quinone formation occurs spontaneously, can be accelerated by metal ions (manganese or iron), and also arises from selected enzyme-catalyzed reactions. Macromolecular damage, combined with increased oxidant stress, may trigger cellular responses that eventually lead to cell death. Reactive quinones have long been known to represent environmental toxicants and, within the context of dopamine metabolism, may also play a role in pathological processes associated with neurodegeneration. The present discussion will review the oxidative metabolism of dopamine and describe experimental evidence suggesting that dopamine quinone may contribute to the cytotoxic and genotoxic potential of this essential neurotransmitter.

Dopamine quinone formation and protein modification associated with the striatal neurotoxicity of methamphetamine: evidence against a role for extracellular dopamine.

Methamphetamine-induced toxicity has been shown to require striatal dopamine and to involve mechanisms associated with oxidative stress. Dopamine is a reactive molecule that can oxidize to form free radicals and reactive quinones. Although this has been suggested to contribute to the mechanism of toxicity, the oxidation of dopamine has never been directly measured after methamphetamine exposure. In this study we sought to determine whether methamphetamine-induced toxicity is associated with the oxidation of dopamine by measuring the binding of dopamine quinones to cysteinyl residues on protein. We observed that administration of neurotoxic doses of methamphetamine to rats resulted in a two- to threefold increase in protein cysteinyl-dopamine in the striatum 2, 4, and 8 hr after treatment. When methamphetamine was administered at an ambient temperature of 5 degreesC, no increase in dopamine oxidation products was observed, and toxicity was prevented. Furthermore, as shown by striatal microdialysis, animals treated with methamphetamine at 5 degreesC showed DA release identical to that of animals treated at room temperature. These data suggest that the toxicity of methamphetamine and the associated increase in dopamine oxidation are not exclusively the result of increases in extracellular dopamine. Because dopamine-induced modifications of protein structure and function may result in cellular toxicity, it is likely that dopamine oxidation contributes to methamphetamine-induced toxicity to dopamine terminals, adding support to the role of dopamine and the evidence of oxidative stress in this lesion model.

Role of endogenous glutathione in the oxidation of dopamine.

Intrastriatal injection of dopamine causes the selective degeneration of tyrosine hydroxylase-containing terminals and an increase in content of cysteinyl-catechols, an index of dopamine oxidation. Both of these effects can be attenuated by coadministration of antioxidants such as glutathione. Therefore, we investigated the effects of decreased endogenous glutathione on the neurotoxic potential of dopamine. We observed that pretreatment with either L-buthionine sulfoximine, a specific inhibitor of glutathione synthesis, or diethyl maleate, which forms adducts with glutathione, caused significant decreases in endogenous glutathione levels at the time of dopamine injection. Pretreatment with L-buthionine sulfoximine potentiated the formation of protein cysteinyl-dopamine after intrastriatal injection of 1.0 micromol of dopamine. We also observed that intrastriatal injection of 1.0 micromol of dopamine decreased striatal glutathione content in all pretreatment conditions. However, injection of a low dose (0.05 micromol of dopamine) caused a decrease in striatal glutathione levels only in the L-buthionine sulfoximine-pretreated rats. Diethyl maleate pretreatment was not effective in potentiating either cysteinyl-catechol formation or glutathione loss after dopamine injection. We conclude that dopamine contributes to cellular oxidative stress and that this can be exacerbated, or at least unmasked, if glutathione synthesis is compromised.

Inhibition of glutamate transport in synaptosomes by dopamine oxidation and reactive oxygen species.

Dopamine can form reactive oxygen species and other reactive metabolites that can modify proteins and other cellular constituents. In this study, we tested the effect of dopamine oxidation products, other generators of reactive oxygen species, and a sulfhydryl modifier on the function of glutamate transporter proteins. We also compared any effects with those on the dopamine transporter, a protein whose function we had previously shown to be inhibited by dopamine oxidation. Preincubation with the generators of reactive oxygen species, ascorbate (0.85 mM) or xanthine (500 microM) plus xanthine oxidase (25 mU/ml), inhibited the uptake of [3H]glutamate (10 microM) into rat striatal synaptosomes (-54 and -74%, respectively). The sulfhydryl-modifying agent N-ethylmaleimide (50-500 microM) also led to a dose-dependent inhibition of [3H]glutamate uptake. Preincubation with dopamine (100 microM) under oxidizing conditions inhibited [3H]glutamate uptake by 25%. Exposure of synaptosomes to increasing amounts of dopamine quinone by enzymatically oxidizing dopamine with tyrosinase (2-50 U/ml) further inhibited [3H]glutamate uptake, an effect prevented by the addition of glutathione. The effects of free radical generators and dopamine oxidation on [3H]glutamate uptake were similar to the effects on [3H]dopamine uptake (250 nM). Our findings suggest that reactive oxygen species and dopamine oxidation products can modify glutamate transport function, which may have implications for neurodegenerative processes such as ischemia, methamphetamine-induced toxicity, and Parkinson's disease.

Characterization of hydrogen peroxide toxicity in cultured rat forebrain neurons.

We investigated the ability of hydrogen peroxide (H2O2) to cause apoptotic cell death in cultured rat forebrain neurons and the potential mechanisms by which oxidative stress triggers delayed neuronal death. H2O2 (25 microM for 5 min) reduced cell viability to 34.5 +/- 8.3% of untreated controls 20 h after exposure, and resulted in a significant proportion of neurons which exhibited apoptotic nuclear morphology. Using single cell fluorescence assays, we measured H2O2-induced changes in DNA strand breaks, 2'7' dichlorofluorescin fluorescence, reduced glutathione, intracellular free Ca2+, and mitochondrial membrane potential. DNA strand breaks in response to H2O2 were not evident immediately following exposure, but were increased 12h and 20h after exposure. Millimolar concentrations of H2O2 caused increases in the fluorescence of the oxidant-sensitive fluorescent dye, 2'7'-dichlorofluorescin. H2O2 treatment decreased reduced glutathione following 30 minutes of exposure using the fluorescent indicator, 5-chloromethylfluorescein diacetate, and increased intraneuronal free Ca2+ levels in a subpopulation of neurons. Mitochondrial membrane potential, measured by rhodamine 123 localization was unaffected by 25 microM H2O2, while higher concentrations of H2O2 (10 or 30 mM) depolarized mitochondria. These studies demonstrate that H2O2 is a potent and effective neurotoxin that produces oxidative stress, as well as apoptotic neuronal death.

Mechanisms of dopamine-induced cell death in cultured rat forebrain neurons: interactions with and differences from glutamate-induced cell death.

Injury to the brain, whether by ischemia or trauma, results in the uncontrolled release of many neurotransmitters, including glutamate and dopamine. Both of these neurotransmitters are neurotoxic in high concentrations, and the oxidative stress caused by reactive oxygen species generation has been implicated in the mechanism of neurotoxicity. In this study, we used cultured rat forebrain neurons to characterize cell death caused by exposure to dopamine and/or glutamate and to investigate potential acute mechanisms of toxicity. Dopamine exposure (250 microM for 2 h) reduced cell viability to 34. 3 +/- 5.5% of untreated control 20 h later and increased the number of neurons with apoptotic morphology. The antioxidant N-acetylcysteine (100 microM) inhibited dopamine-induced toxicity and prevented the covalent binding of dopamine quinones to protein. In contrast, glutamate toxicity lacked the hallmark characteristics of apoptosis. When neurons were exposed successively to sublethal concentrations of dopamine and glutamate, cell viability at 20 h was reduced to 62.3 +/- 5.2% of untreated control. Apoptosis was not evident, and N-acetylcysteine blocked the potentiating effect of dopamine on glutamate-induced toxicity. We used single-cell fluorescence assays to measure changes in intraneuronal glutathione, intraneuronal Ca2+, mitochondrial membrane potential, and DNA integrity as potential acute inducers of neuronal injury. While changes in these parameters could be demonstrated, none were identified as the sole acute inducer of cell death caused by dopamine. In summary, we have characterized a number of neuronal responses to lethal dopamine injury. Also, we have demonstrated that dopamine and glutamate can interact in vitro to potentiate cell death and that the potentiation appears to be induced by oxidative stress.

Loss of dopaminergic neurons in parkinsonism: possible role of reactive dopamine metabolites.

Parkinson's disease affects one out of every 100 people above the age of 55. Its cause is unknown and although the symptoms can be treated, there is no cure. The disease is associated with the selective loss of neurons that contain biogenic amines, and among these it is the dopamine (DA) neurons of the nigrostraital projection that are the most consistently and severely affected (Bernheimer et al., 1973). In this review we discuss the possibility that DA may act as an endogenous neurotoxin, causing the degeneration of the very neurons that release it. We further suggest that although treatments which increase the synthesis and release of DA reduce the symptoms, they also may serve to exacerbate the neurodegenerative process. We propose that the treatments which increase the antioxidant capacity of brain may be protective.

Modification of dopamine transporter function: effect of reactive oxygen species and dopamine.

Dopamine can oxidize to form reactive oxygen species and quinones, and we have previously shown that dopamine quinones bind covalently to cysteinyl residues on striatal proteins. The dopamine transporter is one of the proteins at risk for this modification, because it has a high affinity for dopamine and contains several cysteinyl residues. Therefore, we tested whether dopamine transport in rat striatal synaptosomes could be affected by generators of reactive oxygen species, including dopamine. Uptake of [3H]dopamine (250 nM) was inhibited by ascorbate (0.85 mM; -44%), and this inhibition was prevented by the iron chelator diethylenetriaminepentaacetic acid (1 mM), suggesting that ascorbate was acting as a prooxidant in the presence of iron. Preincubation with xanthine (500 microM) and xanthine oxidase (50 mU/ml) also reduced [3H]dopamine uptake (-76%). Preincubation with dopamine (100 microM) caused a 60% inhibition of subsequent [3H]dopamine uptake. This dopamine-induced inhibition was attenuated by diethylenetriaminepentaacetic acid (1 mM), which can prevent iron-catalyzed oxidation of dopamine during the preincubation, but was unaffected by the monoamine oxidase inhibitor pargyline (10 microM). None of these incubations caused a loss of membrane integrity as indicated by lactate dehydrogenase release. These findings suggest that reactive oxygen species and possibly dopamine quinones can modify dopamine transport function.

Role of oxidation in the neurotoxic effects of intrastriatal dopamine injections.

We have examined the biochemical and histological effects of high concentrations of dopamine (0.05-1.0 micromol) injected into the rat striatum. Twenty-four hours after such injections, the oxidation products of dopamine and dihydroxyphenylacetic acid were detected as both free and protein-bound cysteinyl dopamine and cysteinyl dihydroxyphenylacetic acid. Protein-bound cysteinyl catechols were increased 7- to 20-fold above control tissue levels. By 7 days postinjection, the protein-bound cysteinyl catechols were still detectable, although reduced in concentration, whereas the free forms could no longer be measured. Histological examination of striatum at 7 days revealed a central core of nonspecific damage including neuronal loss and gliosis. This core was surrounded by a region containing a marked reduction in tyrosine hydroxylase immunoreactivity but no apparent loss of serotonin or synaptophysin immunoreactivity. When dopamine was injected with an equimolar concentration of either ascorbic acid or glutathione, the formation of protein-bound cysteinyl catechols was greatly reduced. Moreover, the specific loss of tyrosine hydroxylase immunoreactivity associated with dopamine injections was no longer detectable, although the nonspecific changes in cytoarchitecture were still apparent. Thus, following its oxidation, dopamine in high concentrations binds to protein in the striatum, an event that is correlated with the specific loss of dopaminergic terminals. We suggest that the selective degeneration of dopamine neurons in Parkinson's disease may be caused by an imbalance between the oxidation of dopamine and the availability of antioxidant defenses.

Glutamate induces the production of reactive oxygen species in cultured forebrain neurons following NMDA receptor activation.

Using the fluorescent dye 2',7'-dichlorodihydrofluorescein (DCF-H2) we investigated the role of glutamate in the production of reactive oxygen species (ROS) in cultured neurons from fetal rat forebrain. The addition of an excitotoxic concentration of glutamate (100 microM) produced a generalized decrease in cellular DCF fluorescence accompanied by local areas of increased fluorescence around the margins of the cell body that could be observed within 2-4 min of glutamate exposure. Increases in fluorescence were dependent on NMDA receptor activation and Ca2+ entry and were blocked by the mitochondrial proton ionophore carbonyl cyanide p-trifluoromethoxyphenylhydrazone (FCCP). Additional studies suggested that the generalized decrease in fluorescence was due to intracellular acidification. These studies suggest a critical role for mitochondria in the production of ROS in association with glutamate excitotoxicity, and additionally demonstrate the feasibility of measuring the production of ROS at the level of the single cell.

Estimating hydroxyl radical content in rat brain using systemic and intraventricular salicylate: impact of methamphetamine.

Free radicals have been implicated in the etiology of many neurodegenerative conditions. Yet, because these species are highly reactive and thus short-lived it has been difficult to test these hypotheses. We adapted a method in which hydroxyl radicals are trapped by salicylate in vivo, resulting in the stable and quantifiable products, 2,3-dihydroxybenzoic acid (DHBA) and 2,5-DHBA. After systemic (100 mg/kg i.p.) or intraventricular (4 mumol) administration of salicylate, the amount of DHBA in striatal tissue correlated with tissue levels of salicylate. After systemic salicylate, the ratio of total DHBA to salicylate in neostriatum was at least 10-fold higher than that observed after central salicylate. In addition, systemic salicylate resulted in considerably higher concentrations of 2,3- and 2,5-DHBA in plasma than in brain. Therefore, a large portion of the DHBA present in brain after systemic salicylate may have been formed in the periphery. A neurotoxic regimen of methamphetamine increased the concentration of DHBA in neostriatum after either central or systemic administration of salicylate. The increase in 2,3-DHBA after the central administration of salicylate was significant at 2 h, but not at 4 h, after the last dose of methamphetamine. These results suggest that (1) when assessing specific events in brain, it is preferable to administer salicylate centrally, and (2) neurotoxic doses of methamphetamine increase the hydroxyl radical content in brain in a time-dependent manner.

Enzymatic oxidation of dopamine: the role of prostaglandin H synthase.

An enzyme responsible for the oxidation of dopamine and formation of neuromelanin in brain has not been identified. Prostaglandin H synthase is prominent in brain and possesses peroxidase activity that may cooxidize dopamine to reactive dopamine quinones. This study examined the ability of purified prostaglandin H synthase to catalyze the oxidation of dopamine in vitro. Dopamine oxidation was determined by monitoring the formation of aminochrome and by examining catechol-modified residues on protein present in the reaction mixture. Aminochrome was formed from dopamine in the presence of prostaglandin H synthase, and the reaction rate was dependent on the concentration of substrate and enzyme in the reaction mixture. Both arachidonic acid and hydrogen peroxide could serve as substrates for the prostaglandin H synthase-catalyzed oxidation of dopamine. Indomethacin blocked the reaction when arachidonic acid was used as a substrate, but not when hydrogen peroxide was used. Enzymatically oxidized dopamine covalently bound to protein, as indicated by the presence of cysteinyl-dopamine residues. Binding was significantly reduced in the absence of enzyme or in the presence of antioxidants. These results suggest that the peroxidase activity of prostaglandin H synthase is responsible for catalyzing the oxidation of dopamine to reactive dopamine quinones. It is possible that prostaglandin H synthase is responsible for the oxidation of dopamine and formation of neuromelanin in vivo, which may have implications for the development of Parkinson's disease. Furthermore, drugs such as aspirin that modulate the activity of this enzyme may provide a potential therapeutic approach for the prevention of Parkinson's disease.

Identification of catechol-protein conjugates in neostriatal slices incubated with [3H]dopamine: impact of ascorbic acid and glutathione.

There is evidence to suggest that degeneration of dopaminergic neurons in Parkinson's disease and certain other conditions results from the action of reactive species generated during the oxidation of dopamine. We, therefore, have begun to explore the conditions under which such reactive species are formed. Tissue slices prepared from rat neostriatum were incubated in a standard Krebs bicarbonate buffer for up to 120 min. In the presence of [3H]dopamine (0.01-100 microM), binding of tritium to the acid-insoluble protein fraction was detected. Binding was attenuated by the addition of ascorbate (0.085-0.85 mM) or glutathione (0.01-1.0 mM) to the buffer. Acid hydrolysis of the protein revealed the presence of cysteinyl-dopamine and cysteinyl-dihydroxyphenylacetic acid residues. These results suggest that dopamine oxidizes to form reactive metabolites, presumably quinones, that then bind to nucleophilic sulfhydryl groups on protein cysteinyl residues. The findings further suggest that the extent to which reactive metabolites are formed is determined in part by the balance between the availability of dopamine and the antioxidant environment.