Varenicline rescues nicotine-induced decrease in motivation for sucrose reinforcement.

Varenicline is one of the top medications used for smoking cessation and is often prescribed before termination of nicotine use. The effect of this combined nicotine and varenicline use on the reward system and motivation for primary reinforcement is underexplored. The goal of this study was to assess the effects of nicotine and varenicline on motivation for a food reinforcer. In Experiment 1, we first assessed the responding for sucrose after pretreatment with nicotine (0, 0.1, or 0.4 mg/kg) and varenicline (0.0, 0.1, 1.0 mg/kg) using a behavioral economics approach. The responding for sucrose was then assessed using a progressive ratio schedule of reinforcement after pretreatment with all possible combinations of nicotine and varenicline doses. In Experiment 2, rats were assessed for the consumption of sucrose in home cages after pretreatment with nicotine and varenicline. We found that (a) nicotine decreased economic demand for sucrose, (b) varenicline rescued nicotine-induced reduction in economic demand for sucrose, and (c) history of varenicline treatment predicted responding for sucrose on a progressive ratio schedule of reinforcement where rats with a history of varenicline treatment responded significantly lower for sucrose across nicotine doses than rats that had not been exposed to varenicline. The results of Experiment 2 largely confirmed that nicotine decreases motivation for sucrose using a passive consumption protocol and that varenicline rescues this effect. Overall, these findings suggest that varenicline interacts with the effects of nicotine by restoring nicotine-induced reduction in motivation for appetitive rewards.

Effects of varenicline on subjective craving and relative reinforcing value of cigarettes.

BACKGROUND: Varenicline is an FDA approved medication for the treatment of nicotine dependence. While the efficacy and safety of this medication have been demonstrated, success rates remain low, and efforts to understand mechanisms of efficacy are in progress. A behavioral economics framework is one unique way to examine how demand for a drug changes under different circumstances. Therefore, the current randomized placebo-controlled, cross-over study aimed to examine effects of varenicline on subjective cigarette craving and objective demand for cigarettes measured by a hypothetical behavioral economic task as well as associations between subjective craving and objective demand. METHOD: Non-treatment seeking (n = 37) daily smokers (>10 cigarettes per day) completed a measure of subjective craving for cigarettes and the Cigarette Purchase Task following overnight nicotine abstinence. Participants completed these measures after 10 days on varenicline (1 mg twice per day) and matched placebo. RESULTS: Analyses revealed a significant reduction in subjective craving for cigarettes while on varenicline (p = 0.01), as compared to placebo, and a sex effect such that females exhibited greater craving than males (p = 0.03). However, there were no medication × sex effects (p = 0.84). Analyses of objective demand for cigarettes found varenicline reduced maximum expenditure (Omax) (p = 0.03). Subjective craving was also associated with various indices of demand. CONCLUSION: Results demonstrated varenicline's efficacy in attenuating subjective craving and objective demand for cigarettes and highlight the partial overlap between dimensions of acute drug motivation, namely subjective craving and behavioral economic indices of cigarette demand.

A behavioral economic analysis of the value-enhancing effects of nicotine and varenicline and the role of nicotinic acetylcholine receptors in male and female rats.

Reinforcement value enhancement by nicotine of non-nicotine rewards is believed to partially motivate smoking behavior. Recently, we showed that the value-enhancing effects of nicotine are well characterized by reinforcer demand models and that the value-enhancing effects of the smoking-cessation aid bupropion (Zyban) are distinct from those of nicotine and differ between the sexes. The present study evaluated potential sex differences in the enhancement effects of nicotine and varenicline (Chantix) using a reinforcer demand methodology. The role of α4ß2* and α7 nicotinic acetylcholine receptors (nAChRs) in the enhancing effects of nicotine and varenicline is also evaluated. Male and female rats (n=12/sex) were trained to lever press maintained by sensory reinforcement by visual stimulus (VS) presentations. Changes in the VS value following nicotine and varenicline administration were assessed using an established reinforcer demand approach. Subsequently, the effects of antagonism of α4ß2* and α7 nAChRs on varenicline and nicotine-induced enhancement active lever-pressing were assessed using a progressive ratio schedule. Nicotine and varenicline enhanced VS demand equivalently between the sexes as evaluated by reinforcer demand. However, α4ß2* receptor antagonism attenuated value enhancement by nicotine and varenicline in females, but only of nicotine in males.

Social reinforcement as alternative to sucrose reinforcement is increased by nicotine and methylphenidate in male Fischer-344 rats.

BACKGROUND: Stimulant drugs such as nicotine (NIC) and methylphenidate (MPH) are hypothesized to increase the reinforcing value of sensory stimuli, thus increasing the effectiveness of such reinforcers as alternatives to sucrose reinforcers. METHODS: Inbred Fischer-344 rats (n = 30) were assigned to three groups: saline (SAL; n = 10), nicotine (NIC; n = 10), or methylphenidate (MPH; n = 10). Testing was done in three phases: sucrose only, (SUC), sucrose and drug (SUC/DRUG), and sucrose, drug, and social reinforcement (SUC/DRUG/SOC). During the SUC phase, rats were trained on a progressive ratio 5 (PR5) reinforcement schedule for sucrose (20% solution). In the SUC/DRUG phase, animals were treated with SAL, NIC (0.4 mg/kg, n = 10 SC), or MPH (2.0 mg/kg, n = 10 IP) 30 min prior to testing. In the SUC/DRUG/SOC phase, animals continued receiving drug treatment, and social reinforcement was introduced concurrently with the sucrose reinforcer. The progressive ratio for each reinforcer ran independently of the others. Reinforcing value was measured as break point (BP), the highest number of responses resulting in a reinforcer. RESULTS: SAL-treated animals showed no significant change in sucrose BP. MPH-treated animals showed decreased sucrose BP in the SUC/DRUG phase, with a further reduction in the SUC/DRUG/SOC phase. NIC-treated animals decreased sucrose BP only when a social alternative was offered. CONCLUSION: Both NIC and MPH reduce the sucrose BP in the presence of a social alternative. The decrease in sucrose responding, coupled with increased social responding, suggests that the social alternative acted as an effective alternative reinforcer to sucrose. From a translational perspective, these results suggest that stimulant drugs such as NIC and MPH may increase the effectiveness of treatments that use alternative social reinforcers to decrease eating.

Preclinical abuse liability assessment of ABT-126, an agonist at the α7 nicotinic acetylcholine receptor (nAChR).

ABT-126 is a nicotinic acetylcholine receptor (nAChR) agonist that is selective for the α7 subtype of the receptor. nAChRs are thought to play a role in a variety of neurocognitive processes and have been a pharmacologic target for disorders with cognitive impairment, including schizophrenia and Alzheimer's disease. As part of the preclinical safety package for ABT-126, its potential for abuse was assessed. While the involvement of the α4ß2 subtype of the nicotinic receptor in the addictive properties of nicotine has been demonstrated, the role of the α7 receptor has been studied much less extensively. A number of preclinical assays of abuse potential including open-field, drug discrimination and self-administration were employed in male rats. ABT-126 had modest effects on locomotor activity in the open-field assay. In nicotine and d-amphetamine drug discrimination assays, ABT-126 administration failed to produce appreciable d-amphetamine-like or nicotine-like responding, suggesting that its interoceptive effects are distinct from those of these drugs of abuse. In rats trained to self-administer cocaine, substitution with ABT-126 was similar to substitution with saline, indicating that it lacks reinforcing effects. No evidence of physical dependence was noted following subchronic administration. Overall, these data suggest that ABT-126 has a low potential for abuse. Together with other literature on this drug class, it appears that drugs that selectively activate α7 nAChRs are not likely to result in abuse or dependence.

A systematic assessment of delay discounting in relation to cocaine and nicotine dependence.

Delay discounting is a measure of impulsivity describing how a reinforcer loses value as the delay to its receipt increases. Greater delay discounting is reliably observed among those with different substance use disorders (SUDs) compared to the general population. Nevertheless, the relation between delay discounting and the type and number of substances used remains unclear. The aim of this study was to compare delay discounting across four groups of participants: cocaine- and nicotine-dependent participants, cocaine-dependent only participants, nicotine-dependent only participants, and non-dependent controls. One hundred and seven participants completed a computerized delay discounting task for hypothetical monetary values. Data were fit to Mazur's hyperbolic equation to derive the discounting rate k. Results showed that delay discounting was significantly greater in the cocaine- and nicotine-dependent group, compared to the nicotine-dependent only group, compared to control group. Delay discounting was also greater in the cocaine-dependent only group relative to the nicotine-dependent only and control groups, but no differences were observed between the cocaine- and nicotine-dependent group and the cocaine-dependent only group. This study provides evidence that delay discounting differs depending on the type of SUD but not on the number of SUDs.

A critical examination of best dose analysis for determining cognitive-enhancing potential of drugs: studies with rhesus monkeys and computer simulations.

RATIONALE: Best dose analysis involves identifying the dose associated with the greatest improvement in performance for each subject and comparing performances associated with these individually determined best doses to control performances. OBJECTIVES: The current experiments were conducted to examine whether significant best dose effects might result from the selective analysis of data rather than an actual drug effect. METHODS: Experiment 1 examined the effects of nicotine and methylphenidate on delayed matching-to-sample (DMTS) and self-ordered spatial search (SOSS) performances in rhesus monkeys (DMTS: n = 7; SOSS: n = 6) to determine the validity and reliability of best dose effects. Experiment 2 used Monte Carlo computer simulations to estimate the likelihood of obtaining a significant outcome when the best dose method was applied to randomly generated data sets for which no difference existed. RESULTS: Significant effects were obtained when the best dose analysis was applied to performances from nondrug sessions, and best dose performances were not significantly different from the best nondrug performances. The doses identified as best doses from two nicotine dose-response curve determinations were unrelated, and the improvement associated with the best dose observed during the first dose-response curve determination was not reliable when the dose was administered repeatedly. Finally, there was a high likelihood of obtaining a statistically significant difference when no real difference existed. CONCLUSIONS: Best dose analysis for the identification of potential therapeutic agents should be replaced by single-subject designs.

Investigating the impact of nicotine on executive functions using a novel virtual reality assessment.

AIMS: Nicotine is known to enhance aspects of cognitive functioning in abstinent smokers, but the effects on specific areas of executive functions and in non-smokers are inconclusive. This may be due in part to the poor sensitivity of tests used to assess executive functions. This study used a new virtual reality assessment of executive functions known as JEF (the Jansari assessment of Executive Functions) to address this issue. DESIGN: A 2 × 2 design manipulating group (smokers and never-smokers) and drug [nicotine (4 mg for smokers; 2 mg for never smokers) versus placebo gum]. SETTING: School of Psychology; University of East London. PARTICIPANTS: Seventy-two participants (aged 18-54 years): 36 minimally deprived (2 hours) smokers and 36 never-smokers. MEASUREMENTS: Components of executive function were measured using the virtual reality paradigm JEF, which assesses eight cognitive constructs simultaneously as well as providing an overall performance measure. FINDINGS: Univariate analyses of variance revealed that nicotine improved overall JEF performance, time-based prospective memory and event-based prospective memory in smokers (P < 0.01), but not in never-smokers. Action-based prospective memory was enhanced in both groups (P < 0.01) and never-smokers out-performed smokers on selective thinking and adaptive thinking (P < 0.01). CONCLUSIONS: Executive functioning and prospective memory as aspects of cognitive performance can be enhanced by nicotine gum in smokers who have abstained for as little as 2 hours.

Effects of acute administration of nicotinic and muscarinic cholinergic agonists and antagonists on performance in different cost-benefit decision making tasks in rats.

RATIONALE: Alterations in cost-benefit decision making accompany numerous neuropsychiatric conditions, including schizophrenia, attention deficit hyperactivity disorder, and addiction. Central cholinergic systems have been linked to the etiology and/or treatment of many of these conditions, but little is known about the role of cholinergic signaling in cost-benefit decision making. OBJECTIVES: The goal of these experiments was to determine how cholinergic signaling is involved in cost-benefit decision making, using a behavioral pharmacological approach. METHODS: Male Long-Evans rats were trained in either "probability discounting" or "delay discounting" tasks, in which rats made discrete-trial choices between a small food reward and a large food reward associated with either varying probabilities of omission or varying delays to delivery, respectively. The effects of acute administration of different doses of nicotinic and muscarinic acetylcholine receptor agonists and antagonists were assessed in each task. RESULTS: In the probability discounting task, acute nicotine administration (1.0 mg/kg) significantly increased choice of the large risky reward, and control experiments suggested that this was due to robust nicotine-induced impairments in behavioral flexibility. In the delay discounting task, the muscarinic antagonists scopolamine (0.03, 0.1, and 0.3 mg/kg) and atropine (0.3 mg/kg) both significantly increased choice of the small immediate reward. Neither mecamylamine nor oxotremorine produced reliable effects on either of the decision making tasks. CONCLUSIONS: These data suggest that cholinergic receptors play multiple roles in decision making contexts which include consideration of reward delay or probability. These roles should be considered when targeting these receptors for therapeutic purposes.

Tobacco psychopharmacology and public health policy: it takes a community.

This commentary is based upon the author's lecture given as the 2010 recipient of the award named in honor of Drs. Joseph V. Brady and Charles R. Schuster, given by the Psychopharmacology and Substance Abuse Division (Division 28) of the American Psychological Association (APA). The focus is on the contributions of many behavioral pharmacology researchers who collaborated very much in the spirit of an interactive community dedicated to the common cause of advancing science in service of public health. Division 28 and its members hold a prominent place in this account because, throughout the 1980s and 1990s, the Division was the lead scientific forum for bringing together researchers addressing the behavioral pharmacology of tobacco and nicotine. The commentary provides an overview of how advances utilizing animal and human models of dependence and withdrawal came to inform public health policy and more recently, tobacco product regulation. The commentary also recounts how efforts by the tobacco industry collided with those of nonindustry researchers, including Division 28 members, and how this was taken up in congressional hearings that addressed behavioral pharmacology research on tobacco. The review concludes with an overview of current challenges to behavioral pharmacology researchers to assist in guiding the regulation of tobacco products by the United States Food and Drug Administration and other national regulatory authorities, as well as guiding the implementation of the international tobacco treaty-the World Health Organization Framework Convention on Tobacco Control.

A randomised double-blind crossover trial of the potential analgesic effect of a transdermal nicotine patch in non-smokers based on objective and subjective assessment.

OBJECTIVE: The results of studies of the analgesic effect of nicotine in humans are complex because these studies have included smokers with variable smoking histories. We investigated whether the use of a 17.5 mg transdermal nicotine (TDN) patch decreased the magnitude of pressure pain on the hands of healthy non-smoking volunteers. DESIGN: This was a randomised double-blind crossover trial. A TDN patch or placebo (drug-free bandage) was applied randomly on the anterior chest of non-smoking volunteers 1 h before the experiments. We measured minimum perceived current and pain threshold on the right hand and then evaluated the magnitude of pressure pain by using the Pain Vision PS-2100 (Nipro Co., Osaka, Japan) which helps in objective quantitative assessment of pain magnitude. After estimating minimum perceived current, pressure pain was produced using a combination of 100-g discs and a rod. The rod and the discs weighing 0 (no disc), 200 (two discs), 400 (four discs), 200 (two discs) and 0 g (no disc) were placed consecutively in this order on the right hand and pain threshold was measured. At the same time, volunteers were asked to rate pain on a numerical rating scale (NRS). Minimum perceived current is the current at which the volunteer perceives the first sensation on applying gradually increasing pulsed current. Pain threshold is the compatible electrode current at which the volunteer feels the intensity of pressure pain. Pain degree is calculated as (pain threshold-minimum perceived current)/minimum perceived current × 100. PARTICIPANTS: Forty non-smoking volunteers were enrolled in this study. RESULTS: No significant differences between groups were observed in minimum perceived current, pain threshold, pain degree or NRS. Of the volunteers who received the nicotine patch, four became anorexic and nauseated and two required anti-emetics. CONCLUSION: The nicotine patch had no analgesic effect in non-smoking volunteers.

Zebrafish assessment of cognitive improvement and anxiolysis: filling the gap between in vitro and rodent models for drug development.

Zebrafish can provide a valuable animal model to screen potential cognitive enhancing and anxiolytic drugs. They are economical and can provide a relatively quick indication of possible functional efficacy. In as much as they have a complex nervous system and elaborate behavioral repertoire, zebrafish can provide a good intermediate model between in vitro receptor and cell-based assays and classic mammalian models for drug screening. In addition, the variety of molecular tools available in zebrafish makes them outstanding models for helping to determine the neuromolecular mechanisms for psychoactive drugs. However, to use zebrafish as a translational model we must have validated, sensitive and efficient behavioral tests. In a series of studies, our lab has developed tests of cognitive function and stress response, which are sensitive to drug effects in a similar manner as rodent models and humans for cognitive enhancement and alleviating stress response. In particular, the three-chamber task for learning and memory was shown to be sensitive to the cognitive enhancing effects of nicotine and has been useful in helping to determine neural mechanisms crucial for nicotinic-induced cognitive enhancement. The novel tank diving test was shown to be a valid and efficient test of stress response. It is sensitive to the reduction in stress-related behaviors due to the amxiolytic drugs diazepam and buspirone but not chlordiazepoxide. Nicotine also causes stress alleviating effects which can be interpreted as anxiolytic effects. Zebrafish models of behavioral pharmacology can be useful to efficiently screen test compounds for drug development and can be useful in helping to determine the mechanisms crucial for new therapeutic treatments of neurobehavioral impairments.

Assessment of α7 nicotinic acetylcholine receptor availability in juvenile pig brain with [¹8F]NS10743.

PURPOSE: To conduct a quantitative PET assessment of the specific binding sites in the brain of juvenile pigs for [(18)F]NS10743, a novel diazabicyclononane derivative targeting α7 nicotinic acetylcholine receptors (α7 nAChRs). METHODS: Dynamic PET recordings were made in isoflurane-anaesthetized juvenile pigs during 120 min after administration of [(18)F]NS10743 under baseline conditions (n = 3) and after blocking of the α7 nAChR with NS6740 (3 mg·kg(-1) bolus + 1 mg·kg(-1)·h(-1) continuous infusion; n = 3). Arterial plasma samples were collected for determining the input function of the unmetabolized tracer. Kinetic analysis of regional brain time-radioactivity curves was performed, and parametric maps were calculated relative to arterial input. RESULTS: Plasma [(18)F]NS10743 passed readily into the brain, with peak uptake occurring in α7 nAChR-expressing brain regions such as the colliculi, thalamus, temporal lobe and hippocampus. The highest SUV(max) was approximately 2.3, whereas the lowest uptake was in the olfactory bulb (SUV(max) 1.53 ± 0.32). Administration of NS6740 significantly decreased [(18)F]NS10743 binding late in the emission recording throughout the brain, except in the olfactory bulb, which was therefore chosen as reference region for calculation of BP(ND). The baseline BP(ND) ranged from 0.39 ± 0.08 in the cerebellum to 0.76 ± 0.07 in the temporal lobe. Pretreatment and constant infusion with NS6740 significantly reduced the BP(ND) in regions with high [(18)F]NS10743 binding (temporal lobe -29%, p = 0.01; midbrain: -35%, p = 0.02), without significantly altering the BP(ND) in low binding regions (cerebellum: -16%, p = 0.2). CONCLUSION: This study confirms the potential of [(18)F]NS10743 as a target-specific radiotracer for the molecular imaging of central α7 nAChRs by PET.

Dose-response assay templates for in vitro assessment of resistance to benzimidazole and nicotinic acetylcholine receptor agonist drugs in human hookworms.

With the implementation of mass drug administration programs for the control of human hookworms, there is a need to monitor for the emergence of drug resistance. We have therefore examined in vitro assays for monitoring sensitivity to benzimidazoles (egg hatch assay) and nicotinic acetylcholine receptor agonist drugs (motility and morphology assays), with a view to developing tools for monitoring drug sensitivity in the field. We have performed assays with Necator americanus and Ancylostoma ceylanicum, and combined this with published data on N. americanus and Ancylostoma caninum, to indicate the breadth of the responses of various hookworm species and isolates in these in vitro assays. This has allowed us to generate assay templates covering the known range of responses, with scope to cover any shift in response that may be indicative of resistance. These assays will have immediate applicability in monitoring for the emergence of drug resistance in human hookworm populations.

Nicotine psychopharmacology: policy and regulatory.

Powerful nerve agent, poison, addictive drug, or wonder medicine of the future? Nicotine has had a long and storied history in pharmacology, physiology, public health and, more recently, in regulatory policy initiatives in the United States and internationally. Psychopharmacology research on nicotine and tobacco came to particular prominence in the latter third of the twentieth century with exploration addressing the role of nicotine in tobacco use, the potential categorization of nicotine as an addictive drug, the pharmacological basis for treatment of tobacco addiction, and the perspective of policy developers seeking to reduce the toll of tobacco use. In fact, the 2005 ratification of the World Health Organization's first global health treaty, the Framework Convention on Tobacco Control, provides further impetus for extending the science foundation for tobacco disease control and policy efforts. Implementation of the treaty's provisions will control tobacco use and reduce the 500 million premature deaths projected to occur in the first half of the twenty-first century from tobacco use. Psychopharmacological research on nicotine and tobacco was important in the rationale and development of the treaty. The public health relevance of psychopharmacology research continues to grow with the realization of the potential of nicotine and related drugs to treat or prevent a diverse range of disorders (e.g., Alzheimer's disease, attention deficit hyperactivity disorder, and pain). Although comprehensive review of the research and implications is beyond the scope of this article, the more modest goal of providing insight into the theoretical, clinical, and policy importance of key psychopharmacology research laboratories over the past few decades is attempted.

An assessment of sex differences in nicotine-induced conditioned taste aversions.

Sex differences in taste aversion learning have been reported for a number of different compounds. It is unknown, however, to what degree, if any, such differences exist when nicotine is the aversion-inducing agent. To address this issue, in the present experiment male and female rats were given limited access to saccharin followed by an intraperitoneal (i.p.) injection of either vehicle or nicotine (0.4, 0.8 or 1.2 mg/kg). Although nicotine induced significant taste aversions in both males and females, the aversions were generally weak at all doses tested. There were no sex differences in the acquisition or strength of the aversions induced by nicotine. The vulnerability to drug abuse has been suggested to be a function of the balance of the rewarding and aversive effects of a drug. Given the relatively weak aversions induced in both sexes and the absence of differences between males and females, it is unlikely that the reported sex difference in the self-administration of nicotine is a function of differences in nicotine's aversive effects. The reported difference in the self-administration of nicotine by males and females is more likely a function of differences in the sensitivity to the rewarding effects of the drug.

Personality traits associated with caffeine intake and smoking.

OBJECTIVES: Some studies find a relationship between certain personality traits, as impulsivity or sensation seeking, and caffeine consumption, but these studies do not consider the potential confounding effect of smoking on caffeine intake, a co-occurrence that has been well demonstrated in epidemiological and clinical studies. The main objective of this cross-sectional study was to analyze the association of personality with caffeine intake controlling for the effects of smoking; a secondary objective was to explore the effect of caffeine intake on the previously known relationship between personality and smoking. METHODS: A sample of 498 adults answered a self-questionnaire including socio-demographic variables, and items regarding consumption of tobacco and caffeine. Personality was measured by the Temperament and Character Inventory (TCI-125). We analyzed the association of personality traits with both caffeine intake and smoking, controlling the possible confounding effects of sex, age and each substance with the other one. RESULTS: Logistic regression analyses showed that the temperamental dimension of novelty seeking was associated with heavy caffeine consumption (>200 mg/day) (adjusted OR=2.0; 95% CI: 1.1-3.9), controlling for the effect of smoking. Moreover, novelty seeking was associated with both smoking (adjusted OR=1.8; 95% CI: 1.1-2.9) and heavy smoking (>20 cigarettes/day) (adjusted OR=1.8; 95% CI: 1.0-3.7), after controlling for the effect of caffeine intake. CONCLUSION: Our study offers an epidemiological evidence of the relationship of novelty seeking, considered to be associated with low basal dopaminergic activity, with both nicotine consumption and heavy caffeine intake, two substances that enhance dopaminergic neurotransmission.

Real-time detection of dopamine released from a nerve model cell by an enzyme-catalyzed luminescence method and its application to drug assessment.

A real-time observation of neurotransmitter release from a nerve cell is a useful method for not only neuroscience research, but also assessing of the influence of chemicals, including drugs, on the human nervous system. In this study, a more simple and sensitive method for real-time monitoring of dopamine release from a nerve model cell was developed. Highly sensitive detection of dopamine was performed by using tyramine oxidase for dopamine oxidation, which was followed by a luminol luminescence reaction. This enzyme-catalyzed luminescence method was applied to observe dopamine release from the PC12 cell as a nerve model cell upon stimulation with acetylcholine and an acetylcholine receptor agonist. The results demonstrated that the real-time monitoring of the activation of the PC12 cell was easily performed by this method. This method possessed many advantages, such as high sensitivity, rapid measurement and no pretreatment for cells. It might be applied to drug screening and the assessment of harmful influences of food additives and pesticides on the nerves.

Involvement of nicotinic acetylcholine receptors in suppression of antimicrobial activity and cytokine responses of alveolar macrophages to Legionella pneumophila infection by nicotine.

Although nicotine is thought to be one of the major immunomodulatory components of cigarette smoking, how nicotine alters the host defense of the lung and, in particular, immune responses of alveolar macrophages, which are critical effector cells in the lung defense to infection, is poorly understood. Nicotinic acetylcholine receptors (nAChRs) are the receptor for nicotine and may be involved in the modulation of macrophage function by nicotine. In this study, therefore, nicotine-induced suppression of antimicrobial activity and cytokine responses of alveolar macrophages mediated by nAChRs to Legionella pneumophila, a causative agent for pneumonia, were examined. The murine MH-S alveolar macrophage cell line cells expressed the messages for alpha4 and beta2 subunits of nAChRs, but not alpha7 subunits, determined by RT-PCR. The nicotine treatment of MH-S alveolar macrophages after infection with L. pneumophila significantly enhanced the replication of bacteria in the macrophages and selectively down-regulated the production of IL-6, IL-12, and TNF-alpha, but not IL-10, induced by infection. These effects were completely blocked by a nonselective antagonist, d-tubocurarine, for nAChRs, but not by a selective antagonist, alpha-bungarotoxin, for alpha7-nAChRs. Furthermore, the stimulation of nAChRs with another agonist, 1,1-dimethyl-4-phenylpiperazinium iodide, showed the same effects, which were blocked by the antagonist d-tubocurarine, on the bacterial replication and cytokine regulation with that of nicotine. Thus, the results revealed that nAChRs, the major exogenous ligands of which are nicotine, are involved in the regulation of macrophage immune function by nicotine and may contribute to the cigarette-induced risk factors for respiratory infections in smokers.