Loss of control over the ethanol consumption: differential transcriptional regulation in prefrontal cortex.

Alcohol use disorder (AUD) is a complex multifactorial disease with heritability of ∼50% and corresponds to the state in which the body triggers a reinforcement or reward compulsive behavior due to ethanol consumption, even when faced with negative consequences. Although several studies have shown the impact of high ethanol intake on the prefrontal cortex (PFC) gene expression, few have addressed the relationship between the patterns of gene expression underlying the compulsive behaviour associated with relapsing. In this study, we used a chronic three-bottle free-choice mouse model to investigate the PFC transcriptome in three different groups of mice drinkers: 'Light drinkers' (preference for water throughout the experiment); 'Heavy drinkers' (preference for ethanol with a non-compulsive intake), and 'Inflexible drinkers' (preference for ethanol with a compulsive drinking component). Our aim was to correlate the intake patterns observed in this model with gene expression changes in the PFC, a brain region critical for the development and maintenance of alcohol addiction. We found that the Camk2a gene showed a downregulated profile only in the Inflexible when compared to the Light drinkers group, the Camk2n1 and Pkp2 genes showed an upregulated profile only in the Inflexible drinkers when compared to the Control group, and the Gja1 gene showed an upregulated profile in the Light and Inflexible drinkers when compared to the Control group. These different transcription patterns have been associated to the presence of alcohol, in the Camk2n1 and Gja1 genes; to the amount of ethanol consumed, in the Camk2a gene; and to the loss of control in the alcohol consumption, in the Pkp2 gene. Here, we provide, for the first time, the potential involvement of the Pkp2 gene in the compulsivity and loss of control over the voluntary ethanol consumption.

Inflexible ethanol intake: A putative link with the Lrrk2 pathway.

Alcoholism is a complex multifactorial disorder with a strong genetic influence. Although several studies have shown the impact of high ethanol intake on the striatal gene expression, few have addressed the relationship between the patterns of gene expression underlying the compulsive behaviour associated with the two major concerns in addiction: the excessive drug consumption and relapsing. In this study, we used a chronic three-bottle free-choice murine model to address striatal transcript regulation among animals with different ethanol intakes and preferences: Light Drinkers (preference for water throughout the experiment), Heavy Drinkers (preference for ethanol with a non-compulsive intake) and Inflexible Drinkers (preference for ethanol and simultaneous loss of control over the drug intake). Our aim was to correlate the intake patterns observed in this model with gene expression changes in the striatum, a brain region critical for the development of alcohol addiction. We found that the transcripts of the Lrrk2 gene, which encodes a multifunctional protein with kinase and GTPase activities, is upregulated only in Inflexible Drinkers suggesting, for the first time, that the Lrrk2 pathway plays a major role in the compulsive ethanol intake behaviour of addicted subjects.

Development of a Computer-Based Format for the Alcohol, Smoking, and Substance Involvement Screening Test (ASSIST) With University Students.

BACKGROUND: The Alcohol, Smoking, and Substance Involvement Screening Test (ASSIST) is a reliable and valid tool for the early detection of harmful and hazardous drug use in primary care settings when administered by interview in the general population. In university students, substance use is high, so a reliable and feasible screening instrument is needed. OBJECTIVES: To compare the computer-based ASSIST (ASSISTc) with the interview format (ASSISTi). METHODS: A convenience sample with counterbalanced design was used alternating between the ASSISTi and ASSISTc with 15-day interval. Although this is not a traditional test-retest reliability study, the same statistical analysis was used: intraclass correlations (ICC), kappa (κ), and Cronbach's alpha (α) to compare the two formats. A satisfaction questionnaire was applied immediately after the second session. RESULTS: Both formats were completed by the students (n = 809) over 15 days. The scores of involvement with all substances and with tobacco, alcohol, cannabis, and cocaine obtained with the two formats demonstrated excellent ICC (> .77). The level of agreement was considered substantial for tobacco (κ = .69) and cannabis (κ = .70) and moderate for alcohol (κ = .58). The consistency of the ASSISTc was considered satisfactory (α: .85 for tobacco, .73 for alcohol, and .87 for cannabis). The analysis of satisfaction and feasibility showed that the ASSISTi was easier to understand, but the two formats were considered similar when considering acceptability, ease of responding, and degree of intimidation. CONCLUSIONS/IMPORTANCE: The two formats are acceptable, the scores are comparable, and they can be used interchangeably.

Reduction of ethanol intake by corticotropin-releasing factor receptor-1 antagonist in "heavy-drinking" mice in a free-choice paradigm.

RATIONALE: We hypothesized that the corticotropin-releasing factor (CRF) system is hyperresponsive in animals with high ethanol intake, which exhibits a reduction of ethanol intake when administered with a CRF1 receptor antagonist. METHODS: Outbred Swiss mice were subjected to a long-term, three-bottle, free-choice paradigm (5 and 10 % [v/v] ethanol and water) that consisted of four phases: acquisition (AC; 10 weeks), withdrawal (W; 2 weeks), reexposure (RE; 2 weeks), and quinine-adulteration (AD; 2 weeks). Based on individual ethanol intake, the mice were classified into three groups: A group, preference for ethanol and persistently high consumption during AD phase; B group, preference for ethanol and a reduction of ethanol intake in the AD phase; and C group; preference for water during all phases. A control group only had access to water. CRF1 receptor messenger RNA (mRNA) levels in the amygdala and the effect of the CRF1 receptor antagonist CP-154,526 on ethanol and water intake in the subgroups were studied. RESULTS: CRF1 transcript levels were higher in the B group than in the control group. The highest dose of CP-154,526 reduced ethanol intake and preference, with no changes in water consumption, in the A group compared with vehicle. The B group exhibited a reduction of both ethanol and water intake, with no changes in preference. The C group exhibited no changes in response to the CRF1 antagonist. CONCLUSIONS: CRF1 receptors appear to be involved in ethanol consumption in mice with high ethanol consumption, and CRF system-mediated neuroadaptations depend on drinking profiles.

Reducing substance involvement in college students: a three-arm parallel-group randomized controlled trial of a computer-based intervention.

The prevalence of alcohol and other drug use is high among college students. Reducing their consumption will likely be beneficial for society as a whole. Computer and web-based interventions are promising for providing behaviorally based information. The present study compared the efficacy of three interventions (computerized screening and motivational intervention [ASSIST/MBIc], non-computerized screening and motivational intervention [ASSIST/MBIi], and screening only [control]) in college students in Curitiba, Brazil. A convenience sample of 458 students scored moderate and high risk on the ASSIST. They were then randomized into the three arms of the randomized controlled trial (ASSIST/MBIc, ASSIST/MBIi [interview], and assessment-only [control]) and assessed at baseline and 3 months later. The ASSIST involvement scores decreased at follow-up compared with baseline in the three groups, suggesting that any intervention is better than no intervention. For alcohol, the specific involvement scores decreased to a low level of risk in the three groups and the MBIc group showed a positive outcome compared with control, and the scores for each question were reduced in the two intervention groups compared to baseline. For tobacco, involvement scores decreased in the three groups, but they maintained moderate risk. For marijuana, a small positive effect was observed in the ASSIST/MBIi and control groups. The ASSIST/MBIc may be a good alternative to interview interventions because it is easy to administer, students frequently use such computer-based technologies, and individually tailored content can be delivered in the absence of a counselor.

Development of a self-report format of ASSIST with university students.

The Alcohol, Smoking and Substance Involvement Screening Test (ASSIST) is a reliable and valid tool to early detection in the harmful and hazardous drug use in primary care settings when administered by interview in the general population. As the risk of substance related problems in university students is high, it is necessary to have screening instruments that can be used beyond the health care settings. Thus, we compared a self-report adaptation of ASSIST with the validated interview format in a convenience sample of university students. A counter-balance design was chosen with students alternating between the interview and the self-report formats. Both formats were completed by all students (n=170) over 30 days. The scores for total involvement, tobacco, alcohol, cannabis and cocaine obtained from the two formats demonstrated good intra-class correlation coefficient (ICC >0.60). The agreement assessed by kappa between questions of the two formats was considered moderate for tobacco (0.76) and cannabis (0.69) and discrete for alcohol (0.47). The consistency of the self-report questionnaire was also good to moderate (Cronbach's alpha of 0.90 for tobacco, 0.71 for alcohol, 0.86 for cannabis and 0.89 for cocaine) and showed acceptable sensitivity (66.7-100%) and specificity (83.5-97.1%) for tobacco, alcohol, cannabis and cocaine when compared to the ASSIST interview format (gold standard). The findings suggest that self-report version is as acceptable as the interview and that the scores on the two formats are comparable. However, the participants reported more motivation for change behavior and more concern about substance use when they were interviewed.

Early risky drug use detection in primary healthcare: how does it work in the real world?

Despite effectiveness in research, the efficacy of screening and brief intervention (SBI) for risky substance users is not adequately understood in routine clinical practice. Primary healthcare professionals (n = 103) from three cities in a metropolitan area in Brazil were trained and supervised in SBI and then screened 40 patients. One year later, meetings were held in each city to obtain feedback. Twenty professionals who fulfilled the task (Yes [Y]) and 24 who did not (No [N]) were individually interviewed about their SBI experience. Reports were independently interpreted and codified by two researchers. The Y and N groups reported the same barriers and positive beliefs, but only the Y group reported no negative issues. The present study lasted from 2007 to 2009.

A transcriptional study in mice with different ethanol-drinking profiles: possible involvement of the GABA(B) receptor.

Previous studies have suggested that γ-aminobutyric acid-B (GABA(B)) receptor agonists effectively reduce ethanol intake. The quantification using real-time polymerase chain reaction of Gabbr1 and Gabbr2 mRNA from the prefrontal cortex, hypothalamus, hippocampus, and striatum in mice exposed to an animal model of the addiction developed in our laboratory was performed to evaluate the involvement of the GABA(B) receptor in ethanol consumption. We used outbred, Swiss mice exposed to a three-bottle free-choice model (water, 5% v/v ethanol, and 10% v/v ethanol) that consisted of four phases: acquisition (AC), withdrawal (W), reexposure (RE), and quinine-adulteration (AD). Based on individual ethanol intake, the mice were classified into three groups: "addicted" (A group; preference for ethanol and persistent consumption during all phases), "heavy" (H group; preference for ethanol and a reduction in ethanol intake in the AD phase compared to AC phase), and "light" (L group; preference for water during all phases). In the prefrontal cortex in the A group, we found high Gabbr1 and Gabbr2 transcription levels, with significantly higher Gabbr1 transcription levels compared with the C (ethanol-naive control mice), L, and H groups. In the hippocampus in the A group, Gabbr2 mRNA levels were significantly lower compared with the C, L, and H groups. In the striatum, we found a significant increase in Gabbr1 transcription levels compared with the C, L, and H groups. No differences in Gabbr1 or Gabbr2 transcription levels were observed in the hypothalamus among groups. In summary, Gabbr1 and Gabbr2 transcription levels were altered in cerebral areas related to drug taking only in mice behaviorally classified as "addicted" drinkers, suggesting that these genes may contribute to high and persistent ethanol consumption.

GABA(B) receptor agonist only reduces ethanol drinking in light-drinking mice.

Baclofen, a GABA(B) agonist, reduces ethanol intake in animals and humans, but the contrary or no effect was also reported. Our previous study demonstrated that mice characterized as "loss of control over ethanol intake" had different Gabbr1 and Gabbr2 transcription levels, which express, respectively, the GABA(B1) and GABA(B2) subunits in brain areas related to addictive behavior. In the present study, we tested baclofen on ethanol intake in mice exposed to the free-choice paradigm. Adult male Swiss mice, individually housed, had free access to three bottles: ethanol (5% and 10%) and water. The protocol had four phases: acquisition (AC, 10 weeks), withdrawal (W, 4 cycles during 2 weeks of 2 day-free-choice and 2 day-only-water), reexposure (RE, 2 weeks), and adulteration of ethanol solutions with quinine (AD, 2 weeks). Mice characterized as "loss of control" (A, n=11, preference for ethanol in AC and maintenance of ethanol intake levels in AD), heavy (H, n=11, preference for ethanol in AC and reduction of ethanol intake levels in AD), and light (L, n=16, preference for water in all phases) drinkers were randomly distributed into two subgroups receiving either intraperitoneal injections of all doses of baclofen (1.25, 2.5, and 5.0mg/kg, given each dose twice in consecutive days) or saline, being exposed to free-choice. Fluid consumption was measured 24h later. Baclofen reduced ethanol intake in group L. In group H a reduction compared to AC was observed. Group A maintained their high ethanol intake even after baclofen treatment. Activation of the GABA(B) receptor depends on the precise balance between the GABA(B1) and GABA(B2) subunits, so the disproportionate transcription levels, we reported in group A, could explain this lack of response to baclofen. These data highlight the importance to test baclofen in individuals with different ethanol drinking profiles, including humans.

Magnesium sulfate and sodium valproate block methylphenidate-induced hyperlocomotion, an animal model of mania.

Magnesium sulfate (MgSO4) is used to treat and prevent eclamptic seizures, and several anticonvulsant drugs (e.g., sodium valproate) are clinically effective antimanic drugs. Psychostimulant-induced hyperlocomotion has been proposed as an animal model for the study of antimanic drugs. The present study evaluated the effects of MgSO4 and sodium valproate (as a positive control) on hyperlocomotion induced by methylphenidate in mice. Acute MgSO4 (300-400 mg/kg), but not sodium valproate (100-300 mg/kg), prevented the increase in locomotor activity induced by methylphenidate (5.0 mg/kg). In contrast, repeated treatment (14 days) with valproate (300 mg/kg), but not MgSO4 (400 mg/kg), blocked methylphenidate-induced hyperlocomotion. Thus, acute MgSO4 exerted antimanic-like effects in this animal model.

Behavioral and neurochemical studies in distinct animal models of ethanol's motivational effects.

In the last decades, the goal of creating a unique and complete model of alcohol use and alcoholism has been replaced by a myriad of different animal models, each addressing a specific feature of problematic alcohol consumption. This mini-review highlights selected findings in the field of alcohol abuse and dependence, as found through the use of animal models. There are models (e.g., drinking in the dark, drinking after alcohol adulteration or alcohol deprivation) in which animals self-administer alcohol, that are useful to analyze determinants and consequences of binge drinking, progression from casual to problematic alcohol use and relapse or loss of control over alcohol drinking. In other models (e.g., conditioned place preference, conditioned taste aversion, ethanol-induced behavioral sensitization) alcohol dosing is precisely controlled by the experimenter. These models are useful to study motivational (i.e, appetitive, aversive and negative reinforcing) effects of alcohol and neuroadaptive changes that occur after repeated alcohol exposure. The study of age-related differences in reactivity to alcohol provides yet another avenue for analyzing alcohol's acute and chronic consequences. Ethanol interacts with several neurotransmitter (dopaminergic, glutamatergic, opioidergic and cannabinoid) and neuromodulators and these interactions are involved in the development and maintenance of alcohol self-administration. The findings described in the review, however, indicate a key role of the endogenous opioid system, notably in the mediation of alcohol's postitive rewarding effects. The Review also highlights the need to further assess the inter-relationship between different indices of ethanol's motivational effects as well as their association with alcohol intake and preference.

Trait anxiety and ethanol: anxiolysis in high-anxiety mice and no relation to intake behavior in an addiction model.

Anxiety has been proposed to play a role in the development of alcohol addiction, but the exact mechanisms by which this occurs remain unclear. The present study aimed to verify the relationship between basal anxiety levels, the anxiolytic-like effect of ethanol, and ethanol intake in mice exposed to an addiction model. In one experiment Swiss mice were characterized as high-anxiety (HA), medium-anxiety (MA), or non-anxiety (NA) in the elevated plus maze and then received saline or ethanol 2 g/kg acutely and chronically and were again exposed to the same test. NA mice decreased while MA mice maintained anxiety indices over the test days, regardless of treatment. HA ethanol-treated mice showed an anxiolytic-like effect, both acutely and chronically, while the saline-treated ones maintained their basal anxiety levels. In another experiment HA and MA mice were exposed to an addiction model based on a 3-bottle free-choice paradigm (ethanol 5% and 10%, and water) consisting of four phases: acquisition (10 weeks), withdrawal (W, 2 weeks), reexposure (2 weeks), and quinine-adulteration (2 weeks). HA and MA control mice had access only to water. Mice were characterized as addicted, heavy-drinker and light-drinker [Fachin-Scheit DJ, Ribeiro AF, Pigatto G, Goeldner FO, Boerngen-Lacerda R. Development of a mouse model of ethanol addiction: naltrexone efficacy in reducing consumption but not craving. J Neural Transm 2006;113:1305-21.]. No difference was observed between HA and MA mice in their preference for and intake of ethanol. No correlation was observed between ethanol intake, during any phase, and anxiety indices measured in the basal tests and during the W phase. The differences in anxiety indices between HA and MA groups persisted in the test performed during ethanol withdrawal, suggesting a "trait" anxiety profile. The data suggest that despite the fact that high anxiety trait levels are important for the anxiolytic-like effects of ethanol, they are not a determining factor for high ethanol intake, at least not under these experimental conditions.

Non-motor function of the midbrain dopaminergic neurons.

The roles of the nigrostriatal pathway are far beyond the simple control of motor functions. The tonic release of dopamine in the dorsal and ventral striatum controls the choice of proper actions toward a given environmental situation. In the striatum, a specific action is triggered by a specific stimulus associated with it. When the subject faces a novel and salient stimulus, the phasic release of dopamine allows synaptic plasticity in the cortico-striatal synapses. Neurons of different regions of cortical areas make synapses that converge to the same medium spine neurons of the striatum. The convergent associations form functional units encoding body parts, objects, locations, and symbolic representations of the subject's world. Such units emerge in the striatum in a repetitive manner, like a mosaic of broken mirrors. The phasic release of dopamine allows the association of units to encode an action of the subject directed to an object or location with the outcome of this action. Reinforced stimulus-action-outcome associations will affect future decision making when the same stimulus (object, location, idea) is presented to the subject in the future. In the absence of a minimal amount of striatal dopamine, no action is initiated as seen in Parkinson's disease subjects. The abnormal and improper association of these units leads to the initiation of unpurposeful and sometimes repetitive actions, as those observed in dyskinetic patients. The association of an excessive reinforcement of some actions, like drug consumption, leads to drug addiction. Improper associations of ideas and unpleasant outcomes may be related to traumatic and depressive symptoms common in many diseases, including Parkinson's disease. The same can be said about the learning and memory impairments observed in demented and nondemented Parkinson's disease patients.

The elevated T-maze as a measure of two types of defensive reactions: a factor analysis.

Using factor analysis, we investigated whether the defensive reactions seen in the elevated T-maze measure different behaviours. Rats were submitted to the elevated T-maze followed by the open-field test. Avoidance 1 and 2 loaded on the same factor, while escape 2 and 3 loaded on a second factor. Baseline avoidance did not load together with locomotor activity in the open-field. These results indicate that the elevated T-maze generates two different defensive behaviours.

Tratamento farmacológico do transtorno de ansiedade generalizada: perspectivas futuras / Pharmacological treatment of generalized anxiety disorder: future perspectives

O presente artigo apresenta uma visäo atualizada e ampla do tratamento farmacológico do transtorno de ansiedade generalizada (TAG). Säo revistos os medicamentos com eficácia comprovada em estudos controlados e atualmente disponíveis na clínica (benzodiazepínicos, buspirona, antidepressivos, betabloqueadores, antipsicóticos e extrato de kava-kava). A seguir, baseados nesses dados, propõe-se um algoritmo de tratametno do TAG. Säo apresentadas as principais linhas de pesquisa de novos fármacos ansiolíticos, descrevendo os principais achados clínicos e pré-clínicos