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Reduced expression of glutamate receptors and phosphorylation of CREB are responsible for in vivo Delta9-THC exposure-impaired hippocampal synaptic plasticity.

Chronic use of marijuana impairs synaptic plasticity and cognitive function. However, the molecular mechanisms by which marijuana alters long-term synaptic plasticity are largely unknown. Here, we show that repeated in vivo exposures to Delta9-THC for 7 consecutive days significantly impaired hippocampal long-term potentiation (LTP) of excitatory glutamatergic synaptic transmission. The Delta9-THC exposure-induced decrease in LTP was prevented by pharmacological inhibition or deletion of the cannabinoid 1 receptor (CB1R). To determine the molecular mechanisms underlying Delta9-THC-altered LTP, we targeted expression and function of the glutamate receptors (GluR) and phosphorylation status of cAMP-response element-binding protein (CREB). Chronic in vivo exposure to Delta9-THC produced CB1R-dependent decreases in expression of hippocampal GluR1, NR2A, and NR2B, the ratio of alpha-amino-3-hydroxy-5-methylisoxazole-4-propionate (AMPA)NMDA receptor-gated currents, and phosphorylation of CREB. Our results suggest that reduced expression and function of the GluR subunits and phosphorylation of CREB may underlie the impaired long-term synaptic plasticity induced by repeated in vivo exposure to Delta9-THC.

Early candidacy for differentiation into heterocysts in the filamentous cyanobacterium Anabaena sp. PCC 7120.

The filamentous cyanobacterium Anabaena sp. PCC 7120 fixes dinitrogen facultatively. Upon depletion of combined nitrogen, about 10% of vegetative cells within the filaments differentiate terminally into nitrogen-fixing cells. The heterocyst has been studied as a model system of prokaryotic cell differentiation, with major focus on signal transduction and pattern formation. The fate of heterocyst differentiation is determined at about the eighth hour of induction (point of no return), well before conspicuous morphological or metabolic changes occur. However, little is known about how the initial heterocysts are selected after the induction by nitrogen deprivation. To address this question, we followed the fate of every cells on agar plates after nitrogen deprivation with an interval of 4 h. About 10% of heterocysts were formed without prior division after the start of nitrogen deprivation. The intensity of fluorescence of GFP in the transformants of hetR-gfp increased markedly in the future heterocysts at the fourth hour with respect to other cells. We also noted that the growing filaments consisted of clusters of four consecutive cells that we call quartets. About 75% of initial heterocysts originated from either of the two outer cells of quartets at the start of nitrogen deprivation. These results suggest that the future heterocysts are loosely selected at early times after the start of nitrogen deprivation, before the commitment. Such early candidacy could be explained by different properties of the outer and inner cells of a quartet, but the molecular nature of candidacy remains to be uncovered.

Cannabinoids Delta(9)-tetrahydrocannabinol and cannabidiol differentially inhibit the lipopolysaccharide-activated NF-kappaB and interferon-betaSTAT proinflammatory pathways in BV-2 microglial cells.

Cannabinoids have been shown to exert anti-inflammatory activities in various in vivo and in vitro experimental models as well as ameliorate various inflammatory degenerative diseases. However, the mechanisms of these effects are not completely understood. Using the BV-2 mouse microglial cell line and lipopolysaccharide (LPS) to induce an inflammatory response, we studied the signaling pathways engaged in the anti-inflammatory effects of cannabinoids as well as their influence on the expression of several genes known to be involved in inflammation. We found that the two major cannabinoids present in marijuana, Delta(9)-tetrahydrocannabinol (THC) and cannabidiol (CBD), decrease the production and release of proinflammatory cytokines, including interleukin-1beta, interleukin-6, and interferon (IFN)beta, from LPS-activated microglial cells. The cannabinoid anti-inflammatory action does not seem to involve the CB1 and CB2 cannabinoid receptors or the abn-CBD-sensitive receptors. In addition, we found that THC and CBD act through different, although partially overlapping, mechanisms. CBD, but not THC, reduces the activity of the NF-kappaB pathway, a primary pathway regulating the expression of proinflammatory genes. Moreover, CBD, but not THC, up-regulates the activation of the STAT3 transcription factor, an element of homeostatic mechanism(s) inducing anti-inflammatory events. Following CBD treatment, but less so with THC, we observed a decreased level of mRNA for the Socs3 gene, a main negative regulator of STATs and particularly of STAT3. However, both CBD and THC decreased the activation of the LPS-induced STAT1 transcription factor, a key player in IFNbeta-dependent proinflammatory processes. In summary, our observations show that CBD and THC vary in their effects on the anti-inflammatory pathways, including the NF-kappaB and IFNbeta-dependent pathways.

Discriminative stimulus functions of methanandamide and delta(9)-THC in rats tests with aminoalkylindoles (WIN55,212-2 and AM678) and ethanol.

The aim of the study was to characterize in vivo the aminoalkylindoles WIN55,212-2 (WIN) and AM678 (naphthalen-1-yl(1-pentyl-1H-indol-3-yl)methanone) as cannabinoid receptor (CB(1)R) ligands using drug discrimination. Tests also involved delta(9)-tetrahydrocannabinol (THC) and R-()-methanandamide (mAEA), a metabolically stable analog of the endogenous ligand anandamide, as well as the CB(1)R selective antagonistinverse agonist rimonabant tests with ethanol assessed pharmacological specificity. We used two different drug discriminations (mAEA and THC) allowing us to explore potential differences in CB(1)R activation which could be attributed to variations in their respective CB(1)R signaling mechanisms. There were two concurrently trained groups of rats. One group discriminated between i.p. injected vehicle and 10 mgkg mAEA. The other group was trained to discriminate between vehicle and 1.8 mgkg THC. Dose generalization curves for AM678, WIN55,212-2, THC, and mAEA suggested the following rank order of potency AM678 > WIN55,212-2 > or THC > mAEA in both drug discrimination groups. Challenge by 1 mgkg rimonabant resulted in shifts to the right of the generalization curves for the two aminoalkylindoles (4.4-fold for AM678 and 11.3-fold for WIN in the mAEA group, whereas for the THC group, the corresponding values were 13 and 2.6, respectively), suggesting surmountable antagonism. Ethanol did not generalize in either of the two groups, suggesting pharmacological specificity. Data are congruent with the general observation that there is substantial overlap in the discriminative stimulus effects of CB(1)R ligands across different chemical classes. However, the quantitative differences in the interactions between the two aminoalkylindoles and rimonabant in the two discrimination groups suggest subtle variations in the ligand-receptor activation(s).

Novel approach to data analysis in cocaine-conditioned place preference.

Only a subgroup of human drug users progress from initial drug taking to drug addiction. The learned associations between the effects of the drug and the environment in which it is experienced is an important aspect of the progression to continued drug taking and drug seeking. These associations can be modeled using the conditioned place preference (CPP) paradigm, although no current method of CPP analysis allows for the identification of within-group variability among subjects. In this study, we adapted a criterion method of analysis to separate CPP expressing from non-CPP expressing rats to study more directly within-group variability in the CPP paradigm. Male Sprague-Dawley rats were conditioned with cocaine (5, 10, 20 mgkg) or saline in an unbiased three-chamber CPP apparatus in either a single-trial or four-trial CPP procedure. A classification and regression tree analysis of time spent in the cocaine-paired chamber established a time of 324 s spent in the cocaine-paired chamber as the criterion for cocaine CPP expression. This criterion effectively discriminated control from cocaine-conditioned rats and was reliable for rats trained in both single trial and four-trial CPP procedures. The criterion method showed an enhanced ability to detect effective doses of cocaine in the single-trial CPP procedure and a blockade of CPP expression by MK 212 (0.125 mgkg) treatment in a subgroup of rats. These data support the utility of the criterion analysis as an adjunct to traditional methods that compare group averages in CPP.

Role of water in molecular docking simulations of cytochrome P450 2D6.

Active-site water molecules form an important component in biological systems, facilitating promiscuous binding or an increase in specificity and affinity. Taking water molecules into account in computational approaches to drug design or site-of-metabolism predictions is currently far from straightforward. In this study, the effects of including water molecules in molecular docking simulations of the important metabolic enzyme cytochrome P450 2D6 are investigated. The structure and dynamics of water molecules that are present in the active site simultaneously with a selected substrate are described, and based on this description, water molecules are selected to be included in docking experiments into multiple protein conformations. Apart from the parent substrate, 11 similar and 53 dissimilar substrates are included to investigate the transferability of active-site hydration sites between substrates. The role of water molecules appears to be highly dependent on the protein conformation and the substrate.

Neural and cardiac toxicities associated with 3,4-methylenedioxymethamphetamine (MDMA).

(-)-3,4-Methylenedioxymethamphetamine (MDMA) is a commonly abused illicit drug which affects multiple organ systems. In animals, high-dose administration of MDMA produces deficits in serotonin (5-HT) neurons (e.g., depletion of forebrain 5-HT) that have been viewed as neurotoxicity. Recent data implicate MDMA in the development of valvular heart disease (VHD). The present paper reviews several issues related to MDMA-associated neural and cardiac toxicities. The hypothesis of MDMA neurotoxicity in rats is evaluated in terms of the effects of MDMA on monoamine neurons, the use of scaling methods to extrapolate MDMA doses across species, and functional consequences of MDMA exposure. A potential treatment regimen (l-5-hydroxytryptophan plus carbidopa) for MDMA-associated neural deficits is discussed. The pathogenesis of MDMA-associated VHD is reviewed with specific reference to the role of valvular 5-HT(2B) receptors. We conclude that pharmacological effects of MDMA occur at the same doses in rats and humans. High doses of MDMA that produce 5-HT depletions in rats are associated with tolerance and impaired 5-HT release. Doses of MDMA that fail to deplete 5-HT in rats can cause persistent behavioral dysfunction, suggesting even moderate doses may pose risks. Finally, the MDMA metabolite, 3,4-methylenedioxyamphetamine (MDA), is a potent 5-HT(2B) agonist which could contribute to the increased risk of VHD observed in heavy MDMA users.

Involvement of nicotinic receptors in methamphetamine- and MDMA-induced neurotoxicity pharmacological implications.

During the last years, we have focused on the study of the neurotoxic effects of 3,4-methylenedioxymethamphetamine (MDMA) and methamphetamine (METH) on the central nervous system (CNS) and their pharmacological prevention methods. In the process of this research, we have used a semipurified synaptosomal preparation from striatum of mice or rats as a reliable in vitro model to study reactive oxygen species (ROS) production by these amphetamine derivatives, which is well-correlated with their dopaminergic injury in in vivo models. Using this preparation, we have demonstrated that blockade of alpha7 nicotinic receptors with methyllycaconitine (MLA) prevents ROS production induced by MDMA and METH. Consequently, in vivo, MLA significantly prevents MDMA- and METH-induced neurotoxicity at dopaminergic level (mouse striatum), without affecting hyperthermia induced by these amphetamines. Additionally, when neuroprotection was assayed with memantine (MEM), a dual antagonist of NMDA and alpha7 receptors, an effective neuroprotection was obtained also ahead of serotonergic injury induced by MDMA in rats. MEM also prevents MDMA effect on serotonin transporter functionality and METH effect on dopamine transporter (DAT), suggesting that behavioral effects of these psychostimulants can also be modulated by MEM. Finally, we have demonstrated that MEM prevents the impaired memory function induced by MDMA, and also, using binding studies with radioligands, we have characterized the interaction of these substances with nicotinic receptors. Studies at molecular level showed that both MDMA and METH displaced competitively the binding of radioligands with homomeric alpha7 and heteromeric nicotinic acetylcholine receptors (nAChRs), indicating that they can directly interact with them. In all the cases, MDMA displayed higher affinity than METH and it was higher for heteromeric than for alpha7 subtype. Pre-incubation of differentiated PC12 cells with MDMA or METH induces nAChR upregulation in a concentration- and time-dependent manner, as many nicotinic ligands do, supporting their functional interaction with nAChRs. Such interaction expands the pharmacological profile of amphetamines and can account for some of their effects.

Opposite effect of phencyclidine on activity-regulated cytoskeleton-associated protein (Arc) in juvenile and adult limbic rat brain regions.

The psychotomimetic effect of NMDA antagonists such as phencyclidine (PCP) in humans spurred the hypoglutamatergic theory of schizophrenia. This theory is supported by animal studies demonstrating schizophrenia-like behavioral and molecular changes following PCP administration to adult or neonatal animals. However, schizophrenia is believed to develop in part due to neurodevelopmental dysfunction during adolescence. Therefore, the effects of PCP in juvenile animals may better reflect the pathophysiology of schizophrenia. Here, we compare the effect of PCP (5mgkgday for 5 days) on activity-regulated cytoskeleton-associated protein (Arc) and parvalbumin mRNA expression in juvenile and adult rats. Arc is a marker for excitatory neurotransmission. Parvalbumin is a marker for GABAergic neurotransmission, known to be reduced in postmortem brains of schizophrenics. PCP reduced parvalbumin mRNA expression in the medial prefrontal cortex (mPFC), ventrolateral orbitofrontal cortex (VLO) and shell of the nucleus accumbens (ACCshell) in both juvenile and adult rats. Contrarily, PCP produced opposite effects on Arc mRNA expression in the mPFC, VLO and ACCshell, leading to decreased expression in juvenile and increased expression in adult rats. The differential effect of PCP in juvenile and adult rats may be caused by the immature functional state of the prefrontal cortex in juvenile rats. These results demonstrate differences between the effects of PCP in juvenile and adult rats. The decrease in Arc mRNA in juvenile rats corresponds best with the proposed hypofrontality in schizophrenia, suggesting the merits of using PCP in juvenile animals as a model for schizophrenia, as this would relate better to the typical onset and clinical features of schizophrenia.

An item response theory analysis of DSM-IV criteria for hallucinogen abuse and dependence in adolescents.

This study applied both item response theory (IRT) and multiple indicators-multiple causes (MIMIC) methods to evaluate item-level psychometric properties of diagnostic questions for hallucinogen use disorders (HUDs), differential item functioning (DIF), and predictors of latent HUD. Data were drawn from 2004-2006 National Surveys on Drug Use and Health. Analyses were based on 1548 past-year hallucinogen users aged 12-17 years. Substance use and symptoms were assessed by audio computer-assisted self-interviewing methods. Abuse and dependence criteria empirically were arrayed along a single continuum of severity. All abuse criteria indicated middle-to-high severity on the IRT-defined HUD continuum, while dependence criteria captured a wider range from the lowest (tolerance and time spent) to the highest (taking larger amounts and inability to cut down) severity levels. There was indication of DIF by hallucinogen users age, gender, raceethnicity, and ecstasy use status. Adjusting for DIF, ecstasy users (vs. non-ecstasy hallucinogen users), females (vs. males), and whites (vs. Hispanics) exhibited increased odds of HUD. Symptoms of hallucinogen abuse and dependence empirically do not reflect two discrete conditions in adolescents. Trends and problems related to hallucinogen use among girls and whites should be examined further to inform the designs of effective gender-appropriate and culturally sensitive prevention programs.

Multicenter, double-blind, randomized, placebo-controlled, parallel-group study of the efficacy, safety, and tolerability of THCCBD extract and THC extract in patients with intractable cancer-related pain.

This study compared the efficacy of a tetrahydrocannabinolcannabidiol (THCCBD) extract, a nonopioid analgesic endocannabinoid system modulator, and a THC extract, with placebo, in relieving pain in patients with advanced cancer. In total, 177 patients with cancer pain, who experienced inadequate analgesia despite chronic opioid dosing, entered a two-week, multicenter, double-blind, randomized, placebo-controlled, parallel-group trial. Patients were randomized to THCCBD extract (n 60), THC extract (n 58), or placebo (n 59). The primary analysis of change from baseline in mean pain Numerical Rating Scale (NRS) score was statistically significantly in favor of THCCBD compared with placebo (improvement of -1.37 vs. -0.69), whereas the THC group showed a nonsignificant change (-1.01 vs. -0.69). Twice as many patients taking THCCBD showed a reduction of more than 30% from baseline pain NRS score when compared with placebo (23 43% vs. 12 21%). The associated odds ratio was statistically significant, whereas the number of THC group responders was similar to placebo (12 23% vs. 12 21%) and did not reach statistical significance. There was no change from baseline in median dose of opioid background medication or mean number of doses of breakthrough medication across treatment groups. No significant group differences were found in the NRS sleep quality or nausea scores or the pain control assessment. However, the results from the European Organisation for Research and Treatment of Cancer Quality of Life Cancer Questionnaire showed a worsening in nausea and vomiting with THCCBD compared with placebo (P 0.02), whereas THC had no difference (P 1.0). Most drug-related adverse events were mildmoderate in severity. This study shows that THCCBD extract is efficacious for relief of pain in patients with advanced cancer pain not fully relieved by strong opioids.

Risk profile of male college athletes who use performance-enhancing substances.

There is a general perception that use of performance-enhancing substances (PESs) does not fit the standard profile of substance use. This study sought to determine whether users of PESs report high-risk patterns of alcohol and other drug use and demonstrate risk behaviors associated with problematic substance use. Anonymous self-report questionnaires were administered to a sample of 234 male student athletes. PES users were defined as college athletes who reported past-year use of a broad array of PESs (including stimulants, hormone precursors, and nutritional supplements). Male athlete PES users (n 73) compared with nonusers (n 160) reported more problematic alcohol-use behaviors and more alcohol- and drug-use-related problems. The former compared with the latter was also more likely to report past-year use of tobacco products, marijuana, cocaine, psychedelics, and prescription drugs without a prescription. In addition, PES users demonstrated higher sensation seeking, and greater coping and enhancement motivations for drinking and marijuana use than non-PES users. Although banned PESs are not typically viewed as having a high addiction potential, male athletes who use these drugs may be more likely to participate in other problematic substance-use behaviors. Importantly, the male athletes in this study who reported PES use also participated in substance-use behaviors that can have profound negative effects on athletic performance. More research on the use of PESs in college athletes is needed.

Investigating the role of serotonin in visual orientation processing using an ecstasy (MDMA)-based research model.

A growing body of evidence suggests that regular ecstasy (3,4-methylenedioxymethamphetamine) use causes lasting changes to central serotonergic functioning in humans, including in the occipital lobe. Serotonin may play a role in visual orientation processing, mediated in the occipital lobe, via lateral inhibition. The tilt aftereffect is an illusion apparent following adaptation to stimuli angled 5-50 degrees from vertical and thought to be affected by lateral inhibition between occipital neurons. A recent study identified an enhanced tilt aftereffect among ecstasy users, but only in a subset that were recently abstinent from amphetamines. The current study examined the effects of ecstasy use, cannabis use and their interacting effect on the magnitude of the tilt aftereffect among participants with no recent history of amphetamine consumption. Eleven ecstasy users, 15 cannabis users, 15 ecstasy plus cannabis users and 15 drug-naïve controls were compared on the magnitude of the tilt aftereffect elicited following adaptation to stimuli angled 15, 30, 40 or 60 degrees from vertical. At a 40 degrees adaptation condition, ecstasy users had a greater magnitude of the tilt aftereffect compared to those that had not taken the drug. Additionally, the extent of ecstasy use was positively associated with the magnitude of the tilt aftereffect generated following 15, 30 and 40 degrees adaptation conditions, but not at 60 degrees. Given that lateral inhibition mediates the tilt aftereffect following adaptation to 5-50 degrees , the findings of a relationship between ecstasy use and tilt magnitude at the 15-40 degrees but not 60 degrees adaptation conditions support a role for serotonin in visual orientation processing via lateral inhibition.

Weekly gamma-hydroxybutyrate exposure sensitizes locomotor hyperactivity to low-dose 3,4-methylenedioxymethamphetamine in rats.

Users of the popular party drug 3,4-methylenedioxymethamphetamine (MDMA) sometimes report combining MDMA with gamma-hydroxybutyrate (GHB) to enhance the pleasurable effects of both drugs. However, very few studies have examined the influences of this drug combination. The present study investigated the development of locomotor sensitization in laboratory rats given 7 once-weekly exposures to either MDMA, GHB or their combination (MDMAGHB). The drugs were administered at a high ambient temperature (28 degrees C) to mimic nightclub conditions. MDMA (5 mgkg), given once weekly, produced a progressively greater locomotor and hyperthermic response over time. In contrast, GHB (500 mgkg) administered weekly produced consistent low levels of locomotor activity and few changes in body temperature. Rats receiving the mixture of MDMA (5 mgkg) and GHB (500 mgkg) showed asymptotic levels of sensitized locomotor activity similar to those seen in rats given MDMA alone, but the development of locomotor sensitization was delayed by coadministered GHB. GHB also delayed the development of MDMA-induced hyperthermia. After a washout period of 5 weeks, rats pre-exposed to MDMA, GHB and MDMAGHB showed no hyperactivity when tested drug-free in the context in which they had previously received drugs, but displayed a sensitized locomotor response to a low challenge dose of MDMA (2.5 mgkg). The response to a low dose of methamphetamine (0.5 mgkg) did not differ among groups. Neurochemical analysis using high-performance liquid chromatography revealed few lasting changes in serotonin, dopamine or their metabolites in the striatum or prefrontal cortex of MDMA- or GHB-pre-exposed rats. These results indicate that GHB modulates the locomotor and hyperthermic response to acute MDMA and that pre-exposure to GHB can sensitize the locomotor response to low doses of MDMA.

Interactions between 3,4-methylenedioxymethamphetamine and ethanol in humans and rodents.

Methylenedioxymethamphetamine (MDMA, ecstasy) is a widely used recreational drug, often associated with dance parties. Users self-report euphoria, a sense of well-being and increased feelings of affiliation. In experimental animals, MDMA produces an acute, rapid release of serotonin and, to a lesser extent, dopamine and norepinephrine in the brain. It can also produce a dose-dependent, life-threatening hyperthermia in rodents, primates and humans. Moreover, there is evidence of long-term neurological and psychological effects in heavy users. In rats, MDMA increases the locomotor activity. When used recreationally, MDMA is often taken with other drugs including amphetamine, cannabis, cocaine or ethanol (EtOH). Epidemiological data suggest that MDMA-EtOH is one of the most common combinations. In rats, EtOH potentiates MDMA-induced hyperactivity but may attenuate its hyperthermic effect, depending on the ambient temperature. The possibility that EtOH may modify the pharmacokinetics and pharmadynamics of MDMA is of concern in terms of liability for misuse abuse. In this short review, we focus on the known interactions between MDMA and EtOH in humans and rodents.

The epidemiology of ecstasy use and harms in Australia.

This paper examines the epidemiology of ecstasy use and harm in Australia using multiple data sources. The data included (1) Australian Customs Service 3,4-methylenedioxymethamphetamine (MDMA) detections (2) the National Drug Strategy Household and Australian Secondary Student Alcohol and Drug Surveys (3) data from Australias ecstasy and Related Drugs Reporting System (4) the number of recorded police incidents for ecstasy possession and distribution collated by the N.S.W. Bureau of Crime Statistics and Research (5) the number of calls to the Alcohol and Drug Information Service and Family Drug Support relating to ecstasy (6) the Alcohol and Other Drug Treatment Services National Minimum Dataset on number of treatment episodes for ecstasy, and (7) N.S.W. Division of Analytical Laboratories toxicology data on number of deaths where MDMA was detected. Recent ecstasy use among adults in the general population has increased, whereas among secondary students it has remained low and stable. The patterns of ecstasy consumption among regular ecstasy users have changed over time. Polydrug use and use for extended periods of time (>48 h) remain common among this group. Frequent ecstasy use is associated with a range of risk behaviours and other problems, which tend to be attributed to a number of drugs along with ecstasy. Few ecstasy users present for treatment for problems related to their ecstasy consumption. Messages and interventions to reduce the risks associated with polydrug use and patterns of extended periods of use are clearly warranted. These messages should be delivered outside of traditional health care settings, as few of these users are engaged with such services.

Executive working memory deficits in abstinent ecstasyMDMA users a critical review.

This review examined studies of executive functioning in abstinent ecstasy (3,4-methylenedioxymethamphetamine, MDMA) users on tasks which had been empirically mapped onto updating, shifting, inhibition and accessing long-term memory executive processes. Studies of some aspects of visuospatial memory performance were also included because of the investment of executive resources in such tasks. Thirty-three studies were identified for the review following searches of the Psychinfo and Medline databases. Inclusion criteria were the reporting of new empirical findings from participants drug free at the time of testing, in peer-reviewed journals in the English language. Evidence for ecstasy-related performance deficits was strongest for the updating of verbal material, and for visuospatial memory tasks requiring additional processing beyond storage and retrieval. Such processing suggested that the overall level of executive demand was an important consideration. Executive shifting showed little evidence of ecstasy-related impairment, whilst examination of inhibition and long-term memory access presented an unclear picture. All but one of the studies had a cross-sectional design. Although this is a potential weakness with regard to confounds, the necessity of such designs was acknowledged. Studies were generally aware of the need to control for potential confounds, especially the effects of other drugs, through a mixture of group designs and statistical techniques. It was recommended that future studies of executive functioning in ecstasy users should detail the relationship of the tasks and dependent variables reported to specific executive processes and consider the level of executive demand imposed by such tasks.

Cortisol and 3,4-methylenedioxymethamphetamine neurohormonal aspects of bioenergetic stress in ecstasy users.

3,4-Methylenedioxymethamphetamine (MDMA) can affect both neurotransmitter and neurohormonal activity. This review will debate the role of the metabolic activation hormone cortisol for the psychobiological effects of ecstasyMDMA. The empirical literature on cortisol release following acute MDMA administration and cortisol functioning in drug-free recreational ecstasyMDMA users will be reviewed. This will be followed by an overview of cortisol as a bioenergetic stress neurohormone, and a debate on how it could be modulating the acute and chronic psychobiological effects of MDMA. Cortisol release is increased by stimulatory factors, including physical activity, thermal stress and stimulant drugs. In laboratory studies MDMA leads to an acute cortisol increase of around 150% in sedentary humans. In MDMA-using dance clubbers, the cortisol levels are increased by around 800%, possibly due to the combined factors of stimulant drug, physical exertion and psychosocial stimulation. Regular ecstasyMDMA users also demonstrate changes in baseline cortisol levels and cortisol reactivity, with compromised hypothalamic-pituitary-adrenal activity. Nonpharmacological research has shown how cortisol is important for psychological aspects such as memory, cognition, sleep, impulsivity, depression and neuronal damage. These same functions are often impaired in recreational ecstasyMDMA users, and cortisol may be an important modulatory co-factor. The energizing hormone cortisol is involved in the psychobiology of MDMA, probably via its effects on energy metabolism. Acute cortisol release may potentiate the stimulating effects of MDMA in dance clubbers. Chronically, cortisol may contribute to the variance in functional and structural consequences of repeated ecstasy usage.

Can the severity of dependence scale be usefully applied to ecstasy

Although use of ecstasy (drugs sold as containing 3,4-methylenedioxymethamphetamine) is prevalent, it is typically infrequent, and treatment presentations involving ecstasy as a principal problem drug are relatively rare. Human case reports and animal literature suggest dependence potential, although there may be some unique aspects to this syndrome for ecstasy in comparison to other substances. The Severity of Dependence Scale (SDS) was examined to determine whether this could usefully identify dependent ecstasy consumers. We conducted a cross-sectional survey of 1,658 frequent (at least monthly) ecstasy consumers across Australia, assessing drug use, associated harms and risk behaviours. Dependence was evaluated with the SDS, using a cut-off of > or 4 to identify potential dependence. One fifth of the participants were screened as potentially dependent. These individuals used ecstasy more frequently, in greater amounts, engaged more extensively in risk behaviours and reported greater role interference than other participants. These findings were independent of methamphetamine use or dependence. The underlying structure of the ecstasy SDS was bifactorial. The SDS has demonstrated construct validity as a screening tool to identify ecstasy users at elevated risk of experiencing adverse consequences, including features of dependence. The underlying structure of dependence symptoms differs for ecstasy compared to other drug classes, and some dependent consumers use the drug infrequently. The unique neurotoxic potential and entactogenic effects of ecstasy may require a distinct nosological classification for the experience of dependence associated with the drug.

MDMA self-administration in laboratory animals a summary of the literature and proposal for future research.

The prevalence of 3,4-methylenedioxymethamphetamine (MDMA) use has increased globally and the pattern of consumption has changed considerably. Previously, a subculture of MDMA users was fairly restricted to the dance club scene. More recently, use has spread outside of this subculture and now many users consume MDMA frequently and in large amounts and some meet criteria for drug abuse andor dependence. Because of confounds associated with studying drug users and abusers, animal models have been employed to investigate potential consequences of drug-taking. Among these, self-administration by laboratory animals has been shown to have excellent predictive validity. There have, however, been mixed results with respect to the ability of MDMA to support and maintain self-administration by laboratory animals. This paper reviews the literature on MDMA self-administration in laboratory primates and rodents. Most of the studies in laboratory animals suggest that only low levels of MDMA are self-administered on a daily basis but some have indicated high levels of self-administered MDMA in certain subjects. Differences might be dependent upon the number of test sessions, prior training conditions or other paradigmatic variables. In most cases, MDMA was found to be a lower efficacy reinforcer than other drugs of abuse. It is suggested that the MDMA self-administration develops following a decrease in MDMA-produced serotonin release that occurs with repeated exposure over an extended period of testing. It is hypothesized that the deficit in central serotonin increases the relative MDMA-produced dopamine responses and that this increase in the dopamine response underlies the development and maintenance of MDMA self-administration.

On the mechanisms underlying 3,4-methylenedioxymethamphetamine toxicity the dilemma of the chicken and the egg.

Administration of 3,4-methylenedioxymethamphetamine (MDMA, ecstasy) to various experimental animals has been shown to induce a selective damage to serotonergic axon terminals. While a great consensus appears to exist regarding the causative role of reactive oxygen species (ROS) in the mechanisms underlying MDMA toxicity, the source of free radicals is still a matter of debate. While some authors support dopamine metabolismoxidation inside 5-hydroxytryptamine (5-HT) terminals as the key factor responsible for ROS formation and final 5-HT terminal degeneration, others believe it is MDMA metabolism into pro-oxidant compounds. Although at first sight both hypotheses appear to contend with each other, it may not be the case. This mini-review was therefore undertaken to try to reconcile both hypotheses and to address the dilemma of the causality of MDMA neurotoxicity.

Behavioral, thermal and neurochemical effects of acute and chronic 3,4-methylenedioxymethamphetamine (Ecstasy) self-administration.

3,4-Methylenedioxymethamphetamine (MDMA) is a popular methamphetamine derivative associated with young adults and all-night dance parties. However, the enduring effects of MDMA at voluntary intake levels have not been extensively investigated. In this study, MDMA-influenced behaviors and core temperatures were assessed over the course of 20 daily MDMA self-administration sessions in rats. In vivo microdialysis techniques were used in a subsequent MDMA challenge test session to determine extracellular nucleus accumbens dopamine (NAcc DA) and serotonin (5-HT) levels in MDMA-experienced and naïve animals before and after a self-administered MDMA injection (3.0mgkg, i.v.). During self-administration sessions, gradual and significant increases in MDMA intake and MDMA-stimulated locomotor activity were observed across sessions. Core temperature significantly decreased during initial MDMA sessions, but was unaltered by the last 10 sessions. In the MDMA challenge test, MDMA-naïve rats showed significantly higher NAcc 5-HT responses compared to MDMA-experienced rats, though MDMA experience did not affect the magnitude of NAcc DA response. The overall findings suggest that changes in MDMA-induced responses over the course of increasing levels of drug exposure may reflect the development of tolerance to a number of MDMA effects.

Selective natural kappa opioid and cannabinoid receptor agonists with a potential role in the treatment of gastrointestinal dysfunction.

Salvinorin A is the major active ingredient of Salvia divinorum, a plant used by the Mazatec Indians of Mexico for spiritual and medical purposes. Different preparations from S. divinorum are also used in traditional healing practices to treat gastrointestinal disorders, including diarrhea. The recent extensive research on salvinorin A and other neoclerodane diterpenoids derived from S. divinorum resulted in a large number of reports describing their isolation, synthesis and structure-activity relationship. This review summarizes the present knowledge on salvinorin A and its analogues, with a focus on the effects on gastrointestinal tissues. We furthermore discuss structural changes in salvinorin A that may facilitate future use of its derivatives in human disease.

Sleep deprivation differentially impairs cognitive performance in abstinent methylenedioxymethamphetamine (Ecstasy) users.

Methylenedioxymethamphetamine (MDMA Ecstasy) is a popular recreational drug and brain serotonin (5-HT) neurotoxin. Neuroimaging data indicate that some human MDMA users develop persistent deficits in brain 5-HT neuronal markers. Although the consequences of MDMA-induced 5-HT neurotoxicity are not fully understood, abstinent MDMA users have been found to have subtle cognitive deficits and altered sleep architecture. The present study sought to test the hypothesis that sleep disturbance plays a role in cognitive deficits in MDMA users. Nineteen abstinent MDMA users and 21 control subjects participated in a 5 d inpatient study in a clinical research unit. Baseline sleep quality was measured using the Pittsburgh Sleep Quality Inventory. Cognitive performance was tested three times daily using a computerized cognitive battery. On the third day of admission, subjects began a 40 h sleep deprivation period and continued cognitive testing using the same daily schedule. At baseline, MDMA users performed less accurately than controls on a task of working memory and more impulsively on four of the seven computerized tests. During sleep deprivation, MDMA users, but not controls, became increasingly impulsive, performing more rapidly at the expense of accuracy on tasks of working and short-term memory. Tests of mediation implicated baseline sleep disturbance in the cognitive decline seen during sleep deprivation. These findings are the first to demonstrate that memory problems in MDMA users may be related, at least in part, to sleep disturbance and suggest that cognitive deficits in MDMA users may become more prominent in situations associated with sleep deprivation.

Rapid modulation of spine morphology by the 5-HT2A serotonin receptor through kalirin-7 signaling.

The 5-HT(2A) serotonin receptor is the most abundant serotonin receptor subtype in the cortex and is predominantly expressed in pyramidal neurons. The 5-HT(2A) receptor is a target of several hallucinogens, antipsychotics, anxiolytics, and antidepressants, and it has been associated with several psychiatric disorders, conditions that are also associated with aberrations in dendritic spine morphogenesis. However, the role of 5-HT(2A) receptors in regulating dendritic spine morphogenesis in cortical neurons is unknown. Here we show that the 5-HT(2A) receptor is present in a subset of spines, in addition to dendritic shafts. It colocalizes with PSD-95 and with multiple PDZ protein-1 (MUPP1) in a subset of dendritic spines of rat cortical pyramidal neurons. MUPP1 is enriched in postsynaptic density (PSD) fractions, is targeted to spines in pyramidal neurons, and enhances the localization of 5-HT(2A) receptors to the cell periphery. 5-HT(2A) receptor activation by the 5-HT(2) receptor agonist DOI induced a transient increase in dendritic spine size, as well as phosphorylation of p21-activated kinase (PAK) in cultured cortical neurons. PAK is a downstream target of the neuronal Rac guanine nucleotide exchange factor (RacGEF) kalirin-7 that is important for spine remodeling. Kalirin-7 regulates dendritic spine morphogenesis in neurons but its role in neuromodulator signaling has not been investigated. We show that peptide interference that prevents the localization of kalirin-7 to the postsynaptic density disrupts DOI-induced PAK phosphorylation and spine morphogenesis. These results suggest a potential role for serotonin signaling in modulating spine morphology and kalirin-7s function at cortical synapses.

Brain-derived neurotrophic factor prevents phencyclidine-induced apoptosis in developing brain by parallel activation of both the ERK and PI-3KAkt pathways.

Phencyclidine is an N-methyl d-aspartate receptor (NMDAR) blocker that has been reported to induce neuronal apoptosis during development and schizophrenia-like behaviors in rats later in life. Brain-derived neurotrophic factor (BDNF) has been shown to prevent neuronal death caused by NMDAR blockade, but the precise mechanism is unknown. This study examined the role of the phosphatidylinositol-3 kinase (PI3K)Akt and extracellular signal-regulated kinase (ERK) pathways in BDNF protection of PCP-induced apoptosis in corticostriatal organotypic cultures. It was observed that BDNF inhibited PCP-induced apoptosis in a concentration-dependent fashion. BDNF effectively prevented PCP-induced inhibition of the ERK and PI-3KAkt pathways and suppressed GSK-3beta activation. Blockade of either PI-3KAkt or ERK activation abolished BDNF protection. Western blot analysis revealed that the PI-3K inhibitor LY294002 prevented the stimulating effect of BDNF on the PI-3KAkt pathway, but had no effect on the ERK pathway. Similarly, the ERK inhibitor PD98059 prevented the stimulating effect of BDNF on the ERK pathway, but not the PI-3KAkt pathway. Co-application of LY294002 and PD98059 had no additional effect on BDNF-evoked activation of Akt or ERK. However, concurrent exposure to PD98059 and LY294002 caused much greater inhibition of BDNF-evoked phosphorylation of GSK-3beta at serine 9 than did LY294002 alone. Finally, either BDNF or GSK-3beta inhibition prevented PCP-induced suppression of cyclic-AMP response element binding protein (CREB) phosphorylation. These data demonstrate that the protective effect of BDNF against PCP-induced apoptosis is mediated by parallel activation of the PI-3KAkt and ERK pathways, most likely involves inhibition of GSK-3beta and activation of CREB.

Interaction of drugs of abuse and maintenance treatments with human P-glycoprotein (ABCB1) and breast cancer resistance protein (ABCG2).

Drug interaction with P-glycoprotein (P-gp) and breast cancer resistance protein (BCRP) may influence its tissue disposition including blood-brain barrier transport and result in potent drug-drug interactions. The limited data obtained using in-vitro models indicate that methadone, buprenorphine, and cannabinoids may interact with human P-gp but almost nothing is known about drugs of abuse and BCRP. We used in vitro P-gp and BCRP inhibition flow cytometric assays with hMDR1- and hBCRP-transfected HEK293 cells to test 14 compounds or metabolites frequently involved in addiction, including buprenorphine, norbuprenorphine, methadone, ibogaine, cocaine, cocaethylene, amphetamine, N-methyl-3,4-methylenedioxyamphetamine, 3,4-methylenedioxyamphetamine, nicotine, ketamine, Delta9-tetrahydrocannabinol (THC), naloxone, and morphine. Drugs that in vitro inhibited P-gp or BCRP were tested in hMDR1- and hBCRP-MDCKII bidirectional transport studies. Human P-gp was significantly inhibited in a concentration-dependent manner by norbuprenorphine>buprenorphine>methadone>ibogaine and THC. Similarly, BCRP was inhibited by buprenorphine>norbuprenorphine>ibogaine and THC. None of the other tested compounds inhibited either transporter, even at high concentration (100 microm). Norbuprenorphine (transport efflux ratio approoximately 11) and methadone (transport efflux ratio approoximately 1.9) transport was P-gp-mediated however, with no significant stereo-selectivity regarding methadone enantiomers. BCRP did not transport any of the tested compounds. However, the clinical significance of the interaction of norbuprenorphine with P-gp remains to be evaluated.

Effect of adolescent exposure to WIN 55212-2 on the acquisition and reinstatement of MDMA-induced conditioned place preference.

The present study employs a conditioned place preference procedure (CPP) to examine the effects of exposure to the cannabinoid agonist WIN 55212-2 (WIN) (0.1 and 0.5mgkg) during adolescence on the reinforcing properties of -3,4-methylenedioxymetamphetamine hydrochloride (MDMA) (1.25 and 2.5mgkg) in mice. On postnatal day (PD) 27, animals received a daily injection of the assigned treatment on 5 consecutive days, and three days later the place conditioning procedure was initiated (PD 35). The results suggest that pre-exposure to cannabinoids strengthens the properties of MDMA and favors reinstatement of the craving for the drug, which endorses the gateway hypothesis.

Cancer-induced cachexia a guide for the oncologist.

Cancer-induced cachexia (CIC) is a paraneoplastic syndrome that may account for up to 20% of deaths in cancer patients. Cachexia includes distinct metabolic changes that are the result of an acute-phase response (APR) mounted by the host as a reaction to tumor cells. These changes include increased muscle proteolysis, increased fat lipolysis, and increased hepatic production of acute-phase proteins such as C-reactive protein and fibrinogen. This APR pathogenesis is an important consideration in trying to treat cachectic patients as most therapies do not target the APR and its subsequent metabolic effects. Although there is currently no cure for CIC, the oncologist frequently encounters cachectic patients in practice, and evidence-based management is needed. We review the current data for assessment of starvation and cachexia, providing guidelines for management that include serum markers and functional assessment. In addition, a review of current therapies is provided, including hypercaloric feeding and nutritional intervention to address starvation, as well as data on appetite stimulants such as corticosteroids and megestrol acetate. Experimental therapies are also discussed, including nonsteroidal antiinflammatory drugs, tumor necrosis factor alpha antagonists, tetrahydrocannabinol, growth hormone, ghrelin, oxandrolone, and omega-3 fatty acids.

Caffeine enhances astroglia and microglia reactivity induced by 3,4-methylenedioxymethamphetamine (ecstasy) in mouse brain.

Several reports suggest that 3,4-methylenedioxymethamphetamine (MDMA) induces neurotoxic effects and gliosis. Since recreational use of MDMA is often associated with caffeinated beverages, we investigated whether caffeine interferes with MDMA-induced astroglia and microglia activation, thus facilitating its neurotoxicity. MDMA (4 x 20 mgkg) was acutely administered to mice alone or in combination with caffeine (10 mgkg). CD11b and GFAP immunoreactivity were evaluated as markers of microglia and astroglia activation in the substantia nigra pars-compacta (SNc) and striatum. MDMA was associated with significantly higher CD11b and GFAP immunoreactivity in striatum, whereas only CD11b was significantly higher than vehicle in SNc. Caffeine potentiated the increase in CD11b and GFAP in the striatum but not in the SNc of MDMA-treated mice. The abuse of MDMA is a growing worldwide problem the results of this study suggest that combination of MDMA plus caffeine by increasing glial activation might have harmful health consequences.

Effect of D1-like and D2-like receptor antagonists on methamphetamine and 3,4-methylenedioxymethamphetamine self-administration in rats.

It has been suggested that activation of dopamine D1-like and D2-like receptors contribute equally to the maintenance of drug self-administration. This study compared the contribution of these receptor subtypes to 3,4-methylenedioxymethamphetamine (MDMA) and methamphetamine (MA) self-administration. Effects of pretreatment with the D2-like receptor antagonist, eticlopride (0.0, 0.0125, 0.025 or 0.05 mgkg, intraperitoneal), on responding maintained by several doses of MDMA (0.5, 1.0 and 2.0 mgkginfusion) and MA (0.05, 0.1 and 0.2 mgkginfusion) were determined. As we have published data showing the effects of the D1-like receptor antagonist, SCH23390 (0.0, 0.01 or 0.02 mgkg, subcutaneous), on MDMA self-administration, effects of this dose range on the MA dose-response curve were determined. In our previous study, 0.02 mgkg SCH23390 produced a rightward shift in the MDMA dose response curve, whereas in the present results, this dose decreased responding maintained by most doses of MA. Eticlopride increased the responding maintained by most doses of MDMA but failed to alter MA self-administration. The present results suggest that both D1-like and D2-like receptors contribute to the maintenance of MDMA self-administration, whereas MA self-administration was more sensitive to D1-like receptor blockade.

Modifiable risk factors of ecstasy use risk perception, current dependence, perceived control, and depression.

Risk perception, perceived behavioral control of obtaining ecstasy (PBC-obtaining), current ecstasy dependence, and recent depression have been associated with past ecstasy use, however, their utility in predicting ecstasy use has not been demonstrated. This study aimed to determine whether these four modifiable risk factors could predict ecstasy use after controlling for socio-demographic covariates and recent polydrug use. Data from 601 ecstasy users in the National Institute on Drug Abuse-funded TriCity Study of Club Drug Use, Abuse and Dependence were analyzed using multivariate logistic regression. Participants were interviewed twice within a 2-week period using standardized instruments. Thirteen percent (n 80) of the participants reported using ecstasy between the two interviews. Low risk perception, high PBC-obtaining (an estimated ecstasy procurement time < 24h), and current ecstasy dependence were statistically associated with ecstasy use between the two interviews. Recent depression was not a significant predictor. Despite not being a target predictor, recent polydrug use was also statistically associated with ecstasy use. The present findings may inform the development of interventions targeting ecstasy users.

Morbidity involving the hallucinogenic designer amines MDA and 2C-I.

A case is presented of a 39-year-old woman who suffered severe debilitation because of a hemorrhagic stroke in the context of substance abuse. The patient presented to the emergency room with rapidly diminishing mental status, hypertension, and vasoconstriction her friends provided a history of ingestion of cocaine, 3,4-methylenedioxymethamphetamine (MDMA), and 2C-I, a novel designer amine. A multi-targeted LC-MSMS method for sympathomimetic amines and related drugs in urine detected and quantified 2C-I and MDA, while ruling out MDMA. The cause of the stroke was determined to be an underlying cerebrovascular abnormality called Moyamoya, secondary to substance abuse. In clinical laboratories, gas chromatography-mass spectrometry or liquid chromatography-tandem mass spectrometry (LC-MSMS) confirmation of a positive amphetamine immunoassay is usually directed only towards amphetamine, methamphetamine, MDMA and MDA. This report demonstrates the utility of testing for a wider menu of compounds using LC-MSMS in order to better characterize the prevalence and toxicities of novel amines such as 2C-I.

Single exposure to cocaine or ecstasy induces DNA damage in brain and other organs of mice.

We evaluated the overall genetic damage induced by different doses of cocaine and MDMA (3,4-Methylenedioxymethamphetamine) in several organs. One hour after intraperitoneal drug administration, mice were euthanized peripheral blood, liver and brain were collected, and the cellular suspensions were used for the single cell gel (comet) assay. We determined that all doses of cocaine and MDMA tested were able to induce DNA damage in blood cells. Extensive genotoxic damage was induced by cocaine or MDMA at the highest doses used in liver cells. Brain cells were affected by all doses administrated. These findings demonstrate that cocaine and MDMA are potent genotoxins.

5-HTTLPR polymorphism, mood disorders and MDMA use in a 3-year follow-up study.

A 3-year longitudinal prospective study was conducted to compare the incidence of substance use disorders (SUD) and non-substance use disorders (NSUD) among ecstasy users and two control groups one of cannabis users and the other of non-drug users. The 5-HTTLPR polymorphism related to NSUD was also studied. A total of 94 subjects were included 37 ecstasy users, 23 cannabis users and 34 non-drug users. SUD and NSUD disorders were diagnosed according to the fourth edition of the Diagnostic and Statistical Manual of Mental Health Disorders criteria using the Psychiatric Research Interview for Substance and Mental Disorders. Incidence Rates (IR) are presented. The 5-HTTLPR polymorphism was analyzed. Hardy-Weinberg equilibrium was studied. The results of the study showed that the highest IR of SUD was cannabis abusedependence in both the ecstasy (IR 48.6100 person-year) and cannabis (IR 2.5100 person-year) groups. There were no new cases of SUD in non-drug users at follow-up. The highest IR of NSUD was primary mood disorder in both the ecstasy (IR 4.2100 person-year) and in the non-drug (IR 1.3100 person-year) groups (P < 0.282). There were no new cases of NSUD in the cannabis group at follow-up. 5-HTTLPR polymorphism was associated with lifetime of primary mood disorders in ecstasy group (P 0.018). Ecstasy use was associated with a higher rate of cannabis abusedependence disorders and mood disorders than cannabis use. In the ecstasy users, 5-HTTLPR polymorphism may result in a high vulnerability to primary mood disorders.

Neurotoxicity of drugs of abuse--the case of methylenedioxyamphetamines (MDMA, ecstasy), and amphetamines.

Ecstasy (MDMA, 3,4-methylendioxymethamphetamine) and the stimulants methamphetamine (METH, speed) and amphetamine are popular drugs among young people, particularly in the dance scene. When given in high doses both MDMA and the stimulant amphetamines are clearly neurotoxic in laboratory animals. MDMA causes selective and persistent lesions of central serotonergic nerve terminals, whereas amphetamines damage both the serotonergic and dopaminergic systems. In recent years, the question of ecstasy-induced neurotoxicity and possible functional sequelae has been addressed in several studies in drug users. Despite large methodological problems, the bulk of evidence suggests residual alterations of serotonergic transmission in MDMA users, although at least partial recovery may occur after long-term abstinence. However, functional sequelae may persist even after longer periods of abstinence. To date, the most consistent findings associate subtle cognitive impairments with ecstasy use, particularly with memory. In contrast, studies on possible long-term neurotoxic effects of stimulant use have been relatively scarce. Preliminary evidence suggests that alterations of the dopaminergic system may persist even after years of abstinence from METH, and may be associated with deficits in motor and cognitive performance. In this paper, we will review the literature focusing on human studies. El éxtasis (MDMA, 3,4 metilendioximetanfetamina), y los estimulantes metanfetaminkos (METH, “speed”) y anfetaminicos son drogas frecuentes entre los jóvenes, especialmenie en lugares de baile, Cuando el MDMA los estimulantes anfetamínicos son administradas en alias dosis a animales de laboratorio resultan claramente neurotóxicas. El MDMA produce lesiones selectivas y persistentes de los terminales nerviosos serotoninérgicos centrales, mientras que las anfetaminas dañan tanio los sisiemas serotoninérgicos como dopaminérgicos. En los últimos años la pregunia acerca de la neurotoxicidad inducida por el éxtasis y las posibles secuelas funcionales ha sido tema de algunos estudios con usuarios de drogas, A pesar de los grandes problemas metodológicos, la amplia evidencia sugiere que exisien alteraciones residuales de la transmisión serotoninérgica en usuarios de MDMA, aunque puede conseguirse cieria recuperatión parcial después de una abstinencia prolongada. Sin embargo, las secuelas funcionales pueden persistir aun después de largos périodes de abstinencia. A la fecha, los hallazgos más consisienies asocian los deierioros cognitivos levés, especialmente las alteraciones de memoria con el uso de éxtasis. En contraste, los estudios acerca de los posibles efectos neurotóxicos a largo plazo por el uso de estimulantes han sido relativamente escasos, La evidencia preliminar sugiere que las alteraciones del sistema dopaminérgico pueden persistir aun después de años de abstinencia de METH y pueden asociarse con déficit en el rendimiento motor y cognitivo. En este artículo se revisará la literatura dedicada a estudios en humanos. Lecstasy (MDMA, 3,4-méthylènedioxyméthamphétamine) et les stimulants méthamphêtaminiques (METH, speed) et amphétaminiques sont des drogues courantes chez les jeunes, surtout dans les milieux festifs. Administrées à dose élevée à des animaux de laboratoire, la MDMA et les amphétamines stimulantes sont clairement neurotoxiques, La MDMA provoque des lésions sélectives et persistantes des terminaisons nerveuses centrales sérotoninergiques et les amphétamines lèsent à la fois les systèmes sérotoninergique et dopaminergique. Ces dernières années, plusieurs études ont traité la question de la neurotoxicité de lecstasy et des séquelles éventuelles chez les consommateurs de cette drogue. Malgré dimportants problèmes méthodologiques, lessentiel des arguments sont en faveur de modifications résiduelles de la transmission sérotoninergique chez les consommateurs de MDMA, récupérables en partie après une longue abstinence. Cependant, même après un arrêt prolongé, des séquelles fonctionnelles peuvent persister. Aujourdhui les résultais les plus constants font état de déficits cognitifs subtils avec lecstasy, surtout mnésiques, À lopposé, les études sur les éventuels effets neuroioxiques à long terme de la consommation de stimulant sont relativement rares. Des résultais préliminaires montrent que les modifications du système dopaminergique persistent même après des années dabstinence de METH, persistance qui peut être associée à des déficits des performances motrices et cognitives. Dans cet article, nous passons en revue la littérature centrée sur les études chez lhomme.

5-hydroxytryptamine receptor stimulation of mitochondrial biogenesis.

Mitochondrial dysfunction is both a cause and target of reactive oxygen species during ischemia-reperfusion, drug, and toxicant injury. After injury, renal proximal tubular cells (RPTC) recover mitochondrial function by increasing the expression of the master regulator of mitochondrial biogenesis, peroxisome-proliferator-activated-receptor-gamma-coactivator-1alpha (PGC-1alpha). The goal of this study was to determine whether 5-hydroxytryptamine (5-HT) receptor agonists increase mitochondrial biogenesis and accelerate the recovery of mitochondrial function. Reverse transcription-polymerase chain reaction analysis confirmed the presence of 5-HT2A, 5-HT2B, and 5-HT2C receptor mRNA in RPTC. The 5-HT2 receptor agonist 1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane hydrochloride (DOI 3-10 microM) increased PGC-1alpha levels, expression of mitochondrial proteins ATP synthase beta and NADH dehydrogenase (ubiquinone) 1beta subcomplex 8 (NDUFB8), MitoTracker Red staining intensity, cellular respiration, and ATP levels through a 5-HT receptor and PGC-1alpha-dependent pathway. Similar effects were observed with the 5-HT2 agonist m-chlorophenylpiperazine and were blocked by the 5-HT2 antagonist 8-3-(4-fluorophenoxy) propyl-1-phenyl-1,3,8-triazaspiro4,5decan-4-one (AMI-193). In addition, DOI accelerated the recovery of mitochondrial function after oxidant-induced injury in RPTC. This is the first report to demonstrate 5-HT receptor-mediated mitochondrial biogenesis, and we suggest that 5-HT-agonists may be effective in the treatment of mitochondrial and cell injury.

A fatality from an oral ingestion of methamphetamine.

The case presented is of a 49-year-old white male decedent who admitted to oral ingestion of methamphetamine. He believed he was being followed by the police while walking his daughter to school in the morning and swallowed the 8-ball of meth, which is known to be one-eighth of an ounce or the equivalent of about 3 g. The following autopsy specimens were analyzed for the presence of methamphetamine and amphetamine by gas chromatography-mass spectrometry femoral blood, urine, bile, vitreous fluid, brain, liver, and gastric contents. Blood drawn at the hospital approximately 12 h after ingestion was also analyzed. The methamphetamine concentration in the hospital blood was 3.0 mgL, and the concentration in the femoral blood from autopsy was 30 mgL. Other drugs confirmed included tramadol, lorazepam, and 11-carboxy-Delta(9)-tetrahydrocannabinol. The pathologist ruled the cause of death to be cardiac dysrhythmia due to excited delirium as a result of methamphetamine drug effects. Discussion of the timeline from ingestion to death and the clinical presentation of the decedent are included.

Implications of plasma Delta9-tetrahydrocannabinol, 11-hydroxy-THC, and 11-nor-9-carboxy-THC concentrations in chronic cannabis smokers.

Delta(9)-Tetrahydrocannabinol (THC) is commonly found in toxicological specimens from driving under the influence and accident investigations. Plasma cannabinoid concentrations were determined in 18 long-term heavy cannabis smokers residing on an in-patient research unit for seven days of monitored abstinence. THC, 11-hydroxy-THC, and 11-nor-9-carboxy-THC (THCCOOH) were quantified by two-dimensional gas chromatography-mass spectrometry with cryofocusing. THC concentrations were > 1 ngmL in nine (50.0%) participants (1.2-5.5 ngmL) on abstinence day 7. THCCOOH was detected (2.8-45.6 ngmL) in all participants on study day 7. THC and THCCOOH median percent concentration decreases (n 18) were 39.5% and 72.9% from day 1 to 7, respectively. Most (88.9%) of the participants had at least one specimen with increased THC compared to the previous day. Cannabis use duration and plasma THCCOOH concentrations were positively correlated on days 1-3 (R 0.584-0.610 p 0.007-0.011). There were no significant correlations between THC concentrations > 0.25 ngmL and body mass index on days 1-7 (R -0.234-0.092 p 0.350-0.766). Measurable THC concentrations after seven days of abstinence indicate a potential mechanism for residual neurocognitive impairment observed in chronic cannabis users. THCs presence in plasma for seven days of abstinence suggests its detection may not indicate recent use in daily cannabis users.

Solid-phase extraction and analysis of THC and carboxy-THC from whole blood using a novel fluorinated solid-phase extraction sorbent and fast liquid chromatography-tandem mass spectrometry.

In this study, solid-phase extraction (SPE) is described using a novel fluorinated heptadecafluorotetrahydrodecyl (C(10)H(4)F(17)) phase to isolate THC and its primary metabolite carboxy-THC from whole blood samples. SPE was performed in hydrophobic mode after samples of whole blood were precipitated with acetonitrile. After applying the sample to the SPE column in aqueous phosphate buffer (pH 7), the sorbent was washed with deionized water and phosphate buffer (pH 7) and dried. The SPE column was eluted with a solvent consisting of ethyl acetatehexanes (5050) containing 2% acetic acid. The eluate was collected, evaporated to dryness, and dissolved in mobile phase (50 microL) for analysis by fast liquid chromatography-tandem mass spectrometry in positivenegative multiple reaction monitoring mode. Chromatography was performed in gradient mode employing a C(18) column and a mobile phase consisting of acetontitrile (containing 0.1% formic acid) and 0.1% aqueous formic acid. The total run time for each analysis was less than 5 min. The limits of detectionquantification for this method were determined to be 0.1 and 0.25 ngmL, respectively. The method was found to be linear from 0.25 to 50 ngmL (r(2) >or 0.995). Recoveries of the individual cannabinoids were found to be greater than 85%. In this report, results of authentic samples analyzed for THC and carboxy-THC are reported using this new methodology.

Automated solid-phase extraction-liquid chromatography-tandem mass spectrometry analysis of 11-nor-Delta9-tetrahydrocannabinol-9-carboxylic acid in human urine specimens application to a high-throughput urine analysis laboratory.

An automated solid-phase extraction coupled with liquid chromatography and tandem mass spectrometry (SPE-LC-MS-MS) method for the analysis of 11-nor-Delta(9)-tetrahydrocannabinol-9-carboxylic acid (THC-COOH) in human urine specimens was developed. The method was linear (R(2) 0.9986) to 1000 ngmL with no carryover evidenced at 2000 ngmL. Limits of quantification and detection were found to be 2 ngmL. Interrun precision was evaluated at the 15 ngmL level over nine batches spanning 15 days (n 45). The coefficient of variation (%CV) was found to be 5.5% over the course of the validation. Intrarun precision of a 15 ngmL control (n 5) ranged from 0.58% CV to 7.4% CV for the same set of analytical batches. Interference was tested using (-)-11-hydroxy-Delta(9)-tetrahydrocannabinol, cannabidiol, (-)-Delta(8)-tetrahydrocannabinol, and cannabinol. One hundred and nineteen specimens previously found to contain THC-COOH by a previously validated gas chromatographic mass spectrometry (GC-MS) procedure were compared to the SPE-LC-MS-MS method. Excellent agreement was found (R(2) 0.9925) for the parallel comparison study. The automated SPE procedure eliminates the human factors of specimen handling, extraction, and derivatization, thereby reducing labor costs and rework resulting from human error or technique issues. Additionally, method runtime is greatly reduced (e.g., during parallel studies the SPE-LC-MS-MS instrument was often finished with analysis by the time the technician finished the offline SPE and derivatization procedure prior to the GC-MS analysis).

Urinary MDMA, MDA, HMMA, and HMA excretion following controlled MDMA administration to humans.

3,4-Methylenedioxymethamphetamine (MDMA), or ecstasy, is excreted as unchanged drug, 3,4-methylenedioxyamphetamine (MDA), and free and glucuronidatedsulfated 4-hydroxy-3-methoxymethamphetamine (HMMA), and 4-hydroxy-3-methoxyamphetamine (HMA) metabolites. The aim of this paper is to describe the pattern and timeframe of excretion of MDMA and its metabolites in urine. Placebo, 1.0 mgkg, and 1.6 mgkg oral MDMA doses were administered double-blind to healthy adult MDMA users on a monitored research unit. All urine was collected, aliquots were hydrolyzed, and analytes quantified by gas chromatography-mass spectrometry. Median C(max), T(max), ratios, first and last detection times, and detection rates were determined. Sixteen participants provided 916 urine specimens. After 1.6 mgkg, median C(max) were 21,470 (MDMA), 2229 (MDA), 20,793 (HMMA), and 876 ngmL (HMA) at median T(max) of 13.9, 23.0, 9.2 and 23.3 h. In the first 24 h, 30.2-34.3% total urinary excretion occurred. HMMA last detection exceeded MDMAs by more than 33 h after both doses. Identification of HMMA as well as MDMA increased the ability to identify positive specimens but required hydrolysis. These MDMA, MDA, HMMA, and HMA pharmacokinetic data may be useful for interpreting workplace, drug treatment, criminal justice, and military urine drug tests. Measurement of urinary HMMA provides the longest detection of MDMA exposure yet is not included in routine monitoring procedures.

The variety of ecstasyMDMA users results from the National Epidemiologic Survey on alcohol and related conditions.

This study investigates the potential heterogeneity of ecstasy or MDMA (3,4-methylenedioxy-N-methylamphetamine) users. Data came from the 2001-2002 National Epidemiologic Survey on Alcohol and Related Conditions (NESARC). Latent class analysis (LCA) and multinomial logistic regression procedures were used to identify subtypes of ecstasy users. Approximately 1.6% (n 562) of adult participants (N 43,093) reported lifetime ecstasy use. LCA identified three subtypes of ecstasy users. Class 1 exhibited pervasive use of most drug classes (ecstasy-polydrug users, 37%). Class 2 reported a high rate of use of marijuana and cocaine and a moderate use of amphetamines (ecstasy-marijuana-stimulant users, 29%). Class 3 was characterized by a high rate of use of marijuana and a low use of primarily prescription-type drugs (ecstasy-marijuana users, 34%). Subtypes were distinguished by family income, history of substance abuse treatment, and familial substance abuse. Class 1 exhibited the highest prevalence of disorders related to the use of marijuana (77%), tobacco (66%), amphetamines (36%), opioids (35%), sedatives (31%), and tranquilizers (30%). The recent resurgence in ecstasy use among adults underscores the need to monitor trends in its use.

Sleep impairment in ecstasypolydrug and cannabis-only users.

The present study investigated aspects of sleep quality in ecstasy and cannabis users. Two-hundred and twenty seven participants (117 ecstasypolydrug users, 53 cannabis users and 57 drug naive participants) took part. The participants completed measures of daytime sleepiness, and indicators of sleep quality. The results demonstrated that ecstasypolydrug users viewed themselves as being more evening types and having poorer sleep quality than cannabis users and drug naive participants. They were also more likely to have missed a nights sleep. The reported differences in sleep type may reflect ecstasy-related serotonergic dysfunction resulting in problems with shifting circadian rhythms.

Attenuation of phencyclidine-induced object recognition deficits by the combination of atypical antipsychotic drugs and pimavanserin (ACP 103), a 5-hydroxytryptamine(2A) receptor inverse agonist.

Subchronic administration of the N-methyl-d-aspartate receptor antagonist, phencyclidine (PCP), in rodents has been shown to produce impairment in novel object recognition (NOR), a model of visual learning and memory. We tested the hypothesis that the selective 5-HT(2A) inverse agonists, pimavanserin and (R)-()-alpha-(2,3-dimethoxyphenyl)-1-2-(4-fluorophenylethyl-4-piperidinemethanol (M100907), would potentiate subeffective doses of atypical antipsychotic drugs (APDs) to reverse the NOR deficits. Female rats received vehicle or PCP (2 mgkg b.i.d.) for 7 days, followed by a 7-day washout. Pimavanserin (3 mgkg) or M100907 (1 mgkg) alone, or four atypicial APDs, risperidone (0.05-0.1 mgkg), melperone (1-3 mgkg), olanzapine (1-2 mgkg), or N-desmethylclozapine (1-2 mgkg), and the typical APD, haloperidol (0.05-0.1 mgkg), were administered alone, or in combination with pimavanserin or M100907, before NOR testing. The exploration times of objects during 3-min acquisition and retention trials, separated by a 1-min interval, were compared by analysis of variance. Vehicle-, but not PCP-treated, animals, explored the novel object significantly more than the familiar in the retention trial (p < 0.05-0.01). Pretreatment with the higher doses of the atypical APDs, but not pimavanserin, M100907, or haloperidol alone, reversed the effects of PCP. The effect of risperidone was blocked by haloperidol pretreatment. Coadministration of pimavanserin or M100907, with ineffective doses of the atypical APDs, but not haloperidol, also reversed the PCP-induced deficit in NOR. These results support the importance of 5-hydroxytryptamine(2A) receptor blockade relative to D(2) receptor blockade in the ability of atypicals to ameliorate the effect of subchronic PCP, a putative measure of cognitive dysfunction in schizophrenia.

Intrauterine cannabis exposure affects fetal growth trajectories the Generation R Study.

Cannabis is the most commonly consumed illicit drug among pregnant women. Intrauterine exposure to cannabis may result in risks for the developing fetus. The importance of intrauterine growth on subsequent psychological and behavioral child development has been demonstrated. This study examined the relation between maternal cannabis use and fetal growth until birth in a population-based sample. Approximately 7,452 mothers enrolled during pregnancy and provided information on substance use and fetal growth. Fetal growth was determined using ultrasound measures in early, mid-, and late pregnancy. Additionally, birth weight was assessed. Maternal cannabis use during pregnancy was associated with growth restriction in mid-and late pregnancy and with lower birth weight. This growth reduction was most pronounced for fetuses exposed to continued maternal cannabis use during pregnancy. Fetal weight in cannabis-exposed fetuses showed a growth reduction of -14.44 gweek (95% confidence interval -22.94 to -5.94, p .001) and head circumference (-0.21 mmweek, 95% confidence interval -0.42 to 0.02, p .07), compared with nonexposed fetuses. Maternal cannabis use during pregnancy resulted in more pronounced growth restriction than maternal tobacco use. Paternal cannabis use was not associated with fetal growth restriction. Maternal cannabis use, even for a short period, may be associated with several adverse fetal growth trajectories.

Cannabinoid receptor type 1- and 2-mediated increase in cyclic AMP inhibits T cell receptor-triggered signaling.

The aim of this study was to characterize inhibitory mechanisms on T cell receptor signaling mediated by the cannabinoid receptors CB1 and CB2. Both receptors are coupled to G(io) proteins, which are associated with inhibition of cyclic AMP formation. In human primary and Jurkat T lymphocytes, activation of CB1 by R()-methanandamide, CB2 by JWH015, and both by Delta9-tetrahydrocannabinol induced a short decrease in cyclic AMP lasting less than 1 h. However, this decrease was followed by a massive (up to 10-fold) and sustained (at least up to 48 h) increase in cyclic AMP. Mediated by the cyclic AMP-activated protein kinase A and C-terminal Src kinase, the cannabinoids induced a stable phosphorylation of the inhibitory Tyr-505 of the leukocyte-specific protein tyrosine kinase (Lck). By thus arresting Lck in its inhibited form, the cannabinoids prevented the dephosphorylation of Lck at Tyr-505 in response to T cell receptor activation, which is necessary for the subsequent initiation of T cell receptor signaling. In this way the cannabinoids inhibited the T cell receptor-triggered signaling, i.e. the activation of the zeta-chain-associated protein kinase of 70 kDa, the linker for activation of T cells, MAPK, the induction of interleukin-2, and T cell proliferation. All of the effects of the cannabinoids were blocked by the CB1 and CB2 antagonists AM281 and AM630. These findings help to better understand the immunosuppressive effects of cannabinoids and explain the beneficial effects of these drugs in the treatment of T cell-mediated autoimmune disorders like multiple sclerosis.

Inhibition of the histone demethylase LSD1 blocks alpha-herpesvirus lytic replication and reactivation from latency.

Reversible methylation of histone tails serves as either a positive signal recognized by transcriptional assemblies or a negative signal that result in repression. Invading viral pathogens that depend upon the host cells transcriptional apparatus are also subject to the regulatory impact of chromatin assembly and modifications. Here we show that infection by the alpha-herpesviruses, herpes simplex virus (HSV) and varicella zoster virus (VZV), results in the rapid accumulation of chromatin bearing repressive histone H3 Lys9 methylation. To enable expression of viral immediate early (IE) genes, both viruses use the cellular transcriptional coactivator host cell factor-1 (HCF-1) to recruit the lysine-specific demethylase-1 (LSD1) to the viral immediate early promoters. Depletion of LSD1 or inhibition of its activity with monoamine oxidase inhibitors (MAOIs) results in the accumulation of repressive chromatin and a block to viral gene expression. As HCF-1 is a component of the Set1 and MLL1 histone H3 Lys4 methyltransferase complexes, it thus coordinates modulation of repressive H3 Lys9 methylation levels with addition of activating H3 Lys4 trimethylation marks. Strikingly, MAOIs also block the reactivation of HSV from latency in sensory neurons, indicating that the HCF-1 complex is a crucial component of the reactivation mechanism. The results support pharmaceutical control of histone modifying enzymes as a strategy for controlling herpesvirus infections.

Cannabinoid self-administration attenuates PCP-induced schizophrenia-like symptoms in adult rats.

Although considerable attention has been paid to the relationship between Cannabis use and schizophrenia, there is a significant uncertainty regarding the role of Cannabis consumption in the pathoetiology of the disorder. We investigated the correlation between voluntary cannabinoid consumption and behavioral traits in an animal model of schizophrenia. Male rats were trained to intravenously self-administer the cannabinoid CB1 receptor agonist WIN55,212-2 (WIN 12.5 microgkginfusion) or vehicle they subsequently received acute or chronic-intermittent intraperitoneal administration of the non-competitive N-methyl-d-aspartate receptor antagonist phencyclidine (PCP 2.5mgkg) or saline. We report that WIN self-administration attenuates PCP-induced deficits in (i) prepulse inhibition (PPI) of the acoustic startle reflex, (ii) cognitive skills, and (iii) sociability, suggesting that cannabinoid consumption can ameliorate the schizophrenia-like behavioral alterations caused by PCP. A parallel study performed in animals receiving WIN on a non-voluntary basis (experimenter-given) confirmed an ameliorating effect of cannabinoid administration on the symptoms of schizophrenia induced by PCP.

Altered adult locomotor activity in rats from phencyclidine treatment on postnatal days 7, 9 and 11, but not repeated ketamine treatment on postnatal day 7.

Neonatal ketamine (KET) or phencyclidine (PCP) treatment can trigger apoptotic neurodegeneration in rodents. Previously, we described KET- and PCP-induced altered body weight and home cage, slant board and forelimb hang behaviors in preweaning rats (Boctor et al., 2008). In that study, L-carnitine (LC) attenuated the KET-induced behavioral alterations and body weight decrements. The four subcutaneous (sc) treatment groups were (1) saline (2) 10 mgkg PCP on PNDs 7, 9 and 11 (3) 20 mgkg KET (6 injections one every 2h on PND 7) or (4) a regimen of KET and 250 mgkg LC (KLC) both administered on PND 7, with additional 250 mgkg doses of LC on PNDs 8-11. A portion of each treatment group was evaluated for postweaning behaviors which included grip strength and motor coordination (postnatal days (PNDs) 22 or 71), locomotor sensitization (PND 42), spatial alternation (PNDs 22-70) and residential running wheel activity (PNDs 72-77). On PND 42 or 78, whole and regional brain weights were measured. Grip strength and motor coordination were unaffected at either age by neonatal treatment. On PND 42, neonatally treated KET- or KLC-treated rats responded to a challenge of 5mgkg KET with activity similar to controls that received the same challenge. Neonatal PCP treatment, however, induced significant sensitization to a challenge of 3mgkg PCP on PND 42 relative to controls that received the same challenge, causing increased activity which was especially profound in females. Performance on a continuous spatial alternation task requiring a win-shift, lose-stay strategy appeared unaffected by neonatal KET or KLC treatment. PCP treatment, however, caused significantly increased random responding and shorter choice latencies. In addition, neonatal PCP treatment elevated light and dark period running wheel activity and reduced PND 42 and 78 body and whole brain weights. These findings provide further evidence that PCP treatment on PNDs 7, 9, and 11 causes subtle cognitive deficits and long-term alterations in activity that are unrelated to deficits in grip strength or motor coordination. Further, repeated KET treatment on PND 7 does not appear to result in severe behavioral modifications.

Induction of energy metabolism related enzymes in yeast Saccharomyces cerevisiae exposed to ibogaine is adaptation to acute decrease in ATP energy pool.

Ibogaine has been extensively studied in the last decades in relation to its anti-addictive properties that have been repeatedly reported as being addiction interruptive and craving eliminative. In our previous study we have already demonstrated induction of energy related enzymes in rat brains treated with ibogaine at a dose of 20mgkg i.p. 24 and 72 h prior to proteomic analysis. In this study a model organism yeast Saccharomyces cerevisiae was cultivated with ibogaine in a concentration of 1mgl. Energy metabolism cluster enzymes glyceraldehyde-3-phosphate dehydrogenase, phosphoglycerate kinase, enolase and alcohol dehydrogenase were induced after 5h of exposure. This is a compensation of demonstrated ATP pool decrease after ibogaine. Yeast in a stationary growth phase is an accepted model for studies of housekeeping metabolism of eukaryotes, including humans. Study showed that ibogaines influence on metabolism is neither species nor tissue specific. Effect is not mediated by binding of ibogaine to receptors, as previously described in literature since they are lacking in this model.

Degradation of N-hydroxy-3,4-methylenedioxymethamphetamine in aqueous solution and its prevention.

N-Hydroxy-3,4-methylenedioxymethamphetamine (N-OH-MDMA) is a lesser known psychedelic drug that has recently circulated in the Japanese illicit drug market. From the instability of the similarly structured N-hydroxy-3,4-methylenedioxyamphetamine (N-OH-MDA) in neutral-to-basic aqueous solution, it was presumed that N-OH-MDMA would also degrade in aqueous solution. The aims of this study were (i) investigation of the degradation of N-OH-MDMA in aqueous solution and its prevention, (ii) identification of the degradation products, (iii) determination of the pKa for the conjugate acid of N-OH-MDMA, and (iv) evaluation of liquid-liquid extraction recovery. N-OH-MDA was also included in some of these studies. N-OH-MDMA degraded to 14.9% of initial concentration after 2 h storage in pH 10 buffer solution at 22 degrees C. This degradation was completely inhibited at least for 2 h by addition of L-ascorbic acid, a strong reactive oxygen scavenger. These findings indicate that reactive oxygen species in alkaline solution were involved in N-OH-MDMA degradation. N-OH-MDA, alpha-methyl-(N-methylene)-3,4-methylenedioxybenzeneethanamine and 3,4-methylenedioxyphenyl-2-propanone oxime were found as degradation products of N-OH-MDMA in alkaline solution. The pKa for the conjugate acid of N-OH-MDMA was determined by titration to be 5.52, which was much lower than that reported for 3,4-methylenedioxymethamphetamine (pKa10.38). Excellent recoveries for N-OH-MDMA and N-OH-MDA (>98%) were achieved by extraction with ethyl acetate or chloroform from a basic buffer (pH 10) solution containing 0.1% L-ascorbic acid.

Sertindole improves sub-chronic PCP-induced reversal learning and episodic memory deficits in rodents involvement of 5-HT(6) and 5-HT (2A) receptor mechanisms.

This study examined the efficacy of sertindole in comparison with a selective 5-HT(6) and a 5-HT(2A) receptor antagonist to reverse sub-chronic phencyclidine (PCP)-induced cognitive deficits in female rats. In the first test, adult female hooded Lister rats were trained to perform an operant reversal learning task to 90% criterion. After training, rats were treated with PCP at 2 mgkg (i.p.) or vehicle twice daily for 7 days, followed by 7 days washout. For the second test, novel object recognition (NOR), a separate batch of rats, had the same sub-chronic PCP dosing regime and washout period. In reversal learning, rats were treated acutely with sertindole, the selective 5-HT(2A) receptor antagonist M100.907 or the selective 5-HT(6) receptor antagonist SB-742457. The PCP-induced selective reversal learning deficit was significantly improved by sertindole, M100.907 and SB-742457. Sertindole also significantly improved the sub-chronic PCP-induced deficit in NOR, a test of episodic memory following a 1 min and 1 h inter-trial interval. In vivo binding studies showed that the dose-response relationship for sertindole in this study most closely correlates with affinity for 5-HT(6) receptor in vivo binding in striatum, although contribution from binding to 5-HT(2A) receptors in vivo in cortex may also provide an important mechanism. The efficacies of selective 5-HT(2A) and 5-HT(6) receptor antagonists suggest potential mechanisms mediating the effects of sertindole, which has high affinity for these 5-HT receptor subtypes. The sertindole-induced improvement in cognitive function in this animal model suggests relevance for the management of cognitive deficit symptoms in schizophrenia.

Mice lacking multidrug resistance protein 1a show altered dopaminergic responses to methylenedioxymethamphetamine (MDMA) in striatum.

Multidrug resistance protein 1a (MDR1a) potentiated methylenedioxymethamphetamine (MDMA)-induced decreases of dopamine (DA) and dopamine transport protein in mouse brain one week after MDMA administration. In the present study, we examined if mdr1a wild-type (mdr1a ) and knock-out (mdr1a --) mice differentially handle the acute effects of MDMA on the nigrostriatal DA system 0-24 h following a single drug injection. 3-way ANOVA revealed significant 2-way interactions of strain x time (F (5,152) 32.4, P < 0.001) and strain x dose (F (3,152) 25.8, P < 0.001) on 3,4-dihydroxyphenylacetic acid (DOPAC)DA ratios in mdr1a and -- mice. 0.3-3 h after 10 mgkg MDMA, DOPACDA ratios were increased in mdr1a mice, but decreased 0.3-1 h after MDMA in mdr1a -- mice. Twenty-four hours after 10 mgkg MDMA, DOPACDA ratios were increased 600% in mdr1a mice compared to saline-treated control mice, while in mdr1a -- mice DOPACDA ratios were unchanged. Striatal MDMA and its metabolite, methylenedioxyamphetamine, concentrations by gas chromatography-mass spectrometry were similar in both strains 0.3-4 h after MDMA, discounting the role of MDR1a-facilitated MDMA transport in observed inter-strain differences. Increased DOPACDA turnover in mdr1a mice following MDMA is consistent with the previous report that MDMA neurotoxicity is increased in mdr1a mice. Increased DA turnover via monoamine oxidase in mdr1a vs -- mice might increase exposure to neurotoxic reactive oxygen species.

A pilot study using nabilone for symptomatic treatment in Huntingtons disease.

Pilot study of nabilone in Huntingtons disease (HD). Double-blind, placebo-controlled, cross-over study of nabilone versus placebo. Primary outcome, Unified Huntingtons Disease Rating Scale (UHDRS) total motor score. Secondary measures UHDRS subsections for chorea, cognition and behavior, and neuropsychiatric inventory (NPI). 44 randomized patients received either nabilone (1 or 2 mg) followed by placebo (n 22), or placebo followed by nabilone (n 22). Recruiting was straightforward. Nabilone safe and well tolerated, no psychotic episodes. Assessment of either dose of nabilone versus placebo showed a treatment difference of 0.86 (95% CI -1.8 to 3.52) for total motor score 1.68 (95% CI 0.44 to 2.92) for chorea 3.57 (95% CI -3.41 to 10.55) for UHDRS cognition 4.01 (95% CI -0.11 to 8.13) for UHDRS behavior, and 6.43 (95% CI 0.2 to 12.66) for the NPI. Larger longer RCT of nabilone in HD is feasible and warranted.

Serotonergic mechanisms on breathing modulation in the rat locus coeruleus.

The locus coeruleus (LC) is a noradrenergic nucleus that plays an important role in the ventilatory response to hypercapnia. This nucleus is densely innervated by serotonergic fibers and contains high density of serotonin (5-HT) receptors, including 5-HT(1A) and 5-HT(2). We assessed the possible modulation of respiratory response to hypercapnia by 5-HT, through 5-HT(1A) and 5-HT(2) receptors, in the LC. To this end, we determined the concentrations of 5-HT and its metabolite 5-hydroxyindole-3-acetic acid (5-HIAA) in the LC after hypercapnic exposure. Pulmonary ventilation (VE , plethysmograph) was measured before and after unilateral microinjection (100 nL) of WAY-100635 (5-HT(1A) antagonist, 5.6 and 56 mM), 8-OHDPAT (5-HT(1A7) agonist, 7 and 15 mM), Ketanserin (5-HT(2A) antagonist, 3.7 and 37 mM), or (-)-2,5-dimethoxy-4-iodoamphetaminehydrochloride (DOI 5-HT(2A) agonist, 6.7 and 67 mM) into the LC, followed by a 60-min period of 7% CO(2) exposure. Hypercapnia increased 5-HTIAA levels and 5-HIAA5-HT ratio within the LC. WAY-100635 and 8-OHDPAT intra-LC decreased the hypercapnic ventilatory response due to a lower tidal volume. Ketanserin increased CO(2) drive to breathing and DOI caused the opposite response, both acting on tidal volume. The current results provide evidence of increased 5-HT release during hypercapnia in the LC and that 5-HT presents an inhibitory modulation of the stimulatory role of LC on hypercapnic ventilatory response, acting through postsynaptic 5-HT(2A) receptors in this nucleus. In addition, hypercapnic responses seem to be also regulated by presynaptic 5-HT(1A) receptors in the LC.

DYRK1A, a novel determinant of the methionine-homocysteine cycle in different mouse models overexpressing this Down-syndrome-associated kinase.

Hyperhomocysteinemia, characterized by increased plasma homocysteine level, is associated with an increased risk of atherosclerosis. On the contrary, patients with Down syndrome appear to be protected from the development of atherosclerosis. We previously found a deleterious effect of hyperhomocysteinemia on expression of DYRK1A, a Down-syndrome-associated kinase. As increased expression of DYRK1A and low plasma homocysteine level have been associated with Down syndrome, we aimed to analyze the effect of its over-expression on homocysteine metabolism in mice. Effects of DYRK1A over-expression were examined by biochemical analysis of methionine metabolites, real-time quantitative reverse-transcription polymerase chain reaction, and enzyme activities. We found that over-expression of Dyrk1a increased the hepatic NAD(P)Hquinone oxidoreductase and S-adenosylhomocysteine hydrolase activities, concomitant with decreased level of plasma homocysteine in three mice models overexpressing Dyrk1a. Moreover, these effects were abolished by treatment with harmine, the most potent and specific inhibitor of Dyrk1a. The increased NAD(P)Hquinone oxidoreductase and S-adenosylhomocysteine hydrolase activities were also found in lymphoblastoid cell lines from patients with Down syndrome. Our results might give clues to understand the protective effect of Down syndrome against vascular defect through a decrease of homocysteine level by DYRK1A over-expression. They reveal a link between the Dyrk1a signaling pathway and the homocysteine cycle.

Administration of neurotoxic doses of MDMA reduces sensitivity to ethanol and increases GAT-1 immunoreactivity in mice striatum.

Mice with reduced dopamine activity following neurotoxic doses of 3,4-methylenedioxymethamphetamine (MDMA, ecstasy) consume more ethanol (EtOH) and show greater preference for EtOH. In keeping with human studies and other animal models where alcohol consumption and preference are also high, MDMA treatment will reduce sensitivity to certain physiological effects of EtOH. We have examined the sensitivity to the acute effects of EtOH in MDMA-lesioned mice and the effects of EtOH on striatal gamma-aminobutyric acid (GABA) accumulation and expression of GABA subtype-1 transporter (GAT-1). C57BL6J mice were injected with neurotoxic MDMA (30 mgkg, three times, every 3 h, i.p.). Seven days later, mice were given EtOH (3 gkg, i.p.) to determine the loss of righting response and the development of rapid tolerance to the hypothermic effect of EtOH. The effect of EtOH on the striatal accumulation of GABA after inhibiting GABA transaminase and on GAT-1 immunoreactivity was also determined. Mice pre-treated with a neurotoxic dose of MDMA were less sensitive to the sedative-hypnotic effect of acute EtOH and exhibited alterations in the development of rapid tolerance to the hypothermic effect of EtOH. These animals showed an increase in striatal GAT-1 immunoreactivity. EtOH reduced GABA concentration in the striatum of non-lesioned mice, an effect not observed in MDMA-lesioned mice. These findings indicate that mice with a MDMA-induced dopaminergic lesion show increased expression of striatal GAT-1 that may contribute to the lower sensitivity to EtOH-induced sedative effects and the resistance to the development of rapid tolerance to hypothermia produced by EtOH.

Acute SSRI-induced anxiogenic and brain metabolic effects are attenuated 6 months after initial MDMA-induced depletion.

To assess the functional state of the serotonergic system, the acute behavioural and brain metabolic effect of SSRI antidepressants were studied during the recovery period after MDMA-induced neuronal damage. The effects of the SSRI fluoxetine and the serotonin receptor agonist meta-chloro-phenylpiperazine (m-CPP) were investigated in the social interaction test in Dark Agouti rats, 6 months after treatment with a single dose of MDMA (15 or 30 mg kg(-1), i.p.). At earlier time points these doses of MDMA have been shown to cause 30-60% loss in axonal densities in several brain regions. Densities of the serotonergic axons were assessed using serotonin-transporter and tryptophan-hydroxylase immunohistochemistry. In a parallel group of animals, brain function was examined following an acute challenge with either fluoxetine or citalopram, using 2-deoxyglucose autoradiographic imaging. Six months after MDMA treatment the densities of serotonergic axons were decreased in only a few brain areas including hippocampus and thalamus. Basal anxiety was unaltered in MDMA-treated animals. However, the acute anxiogenic effects of fluoxetine, but not m-CPP, were attenuated in animals pretreated with MDMA. The metabolic response to both citalopram and fluoxetine was normal in most of the brain areas examined with the exception of ventromedial thalamus and hippocampal sub-fields where the response was attenuated. These data provide evidence that 6 months after MDMA-induced damage serotonergic axons show recovery in most brain areas, but serotonergic functions to challenges with SSRIs including anxiety and aggression remain altered.

Acute nicotine and phencyclidine increase locomotor activity of the guinea pig with attenuated potencies relative to their effects on rat or mouse.

Behavioral assays of the responses to psychomotor stimulants can be used to model certain aspects of CNS pathologies such as psychosis and addiction. However, species-dependent differences in the effects of neuromodulators in these assays can confound the interpretation of the results. The goal of this study was to determine the utility of the guinea pig as a model for assessing the behavioral actions of nicotinic receptor agonists and NMDA receptor antagonists. In the present study, the locomotor activity of adult male guinea pigs was measured, prior to and following an acute injection of nicotine, MK-801 or phencyclidine. Each animal received a single dose of the drug. Nicotine produced a dose-dependent increase in activity with an ED(50) of 1.5mgkg. Phencyclidine also increased activity, with an ED(50) of 3.4 mgkg. Nicotine produced increases in locomotion in all individual subjects tested, whereas at the maximally-effective dose of phencyclidine, only a fraction of the animals had locomotor activation. There was no change in activity in response to a single dose of MK-801 (0.5mgkg). Haloperidol had a significant inhibitory effect on locomotor activity independent of the stimulant administered. Thus, both phencyclidine and nicotine are psychomotor stimulants when given to guinea pigs, although the intensity of the response and the potencies of these drugs are lower than in mice or rats under otherwise similar conditions.

Involvement of cannabinoid-1 and cannabinoid-2 receptors in septic ileus.

BACKGROUND Cannabinoid (CB) receptors are involved in the regulation of gastrointestinal (GI) motility under physiological and pathophysiological conditions. We aimed to characterize the possible influence of CB(1) and CB(2) receptors on motility impairment in a model of septic ileus. METHODS Lipopolysaccharide (LPS) injections were used to mimic pathophysiological features of septic ileus. Spontaneous jejunal myoelectrical activity was measured in rats in vivo, and upper GI transit was measured in vivo by gavaging of a charcoal marker into the stomach of mice, in absence or presence of LPS, and CB(1) and CB(2) receptor agonists and antagonists. Tumour necrosis factor (TNF)-alpha and interleukin (IL)-6 levels were measured using enzyme-linked immunosorbent assay. Histology was performed with haematoxylin-eosin staining. KEY RESULTS Lipopolysaccharide treatment significantly reduced amplitude and frequency of myoelectric spiking activity and GI transit in vivo in a dose-dependent manner. TNF-alpha and IL-6 were increased in LPS-treated animals and histology showed oedema and cell infiltration. Both, the CB(1) agonist HU210 and the CB(2) agonist JWH133 reduced myoelectrical activity whereas the CB(1) antagonist AM251 caused an increase of myoelectrical activity. Pretreatment with AM251 or AM630 prevented against LPS-induced reduction of myoelectrical activity, and also against the delay of GI transit during septic ileus in vivo. CONCLUSIONS INFERENCES The LPS model of septic ileus impairs jejunal myoelectrical activity and delays GI transit in vivo. Antagonists at the CB(1) receptor or the CB(2) receptor prevent the delay of GI transit and thus may be powerful tools in the future treatment of septic ileus.

Adverse health effects of non-medical cannabis use.

For over two decades, cannabis, commonly known as marijuana, has been the most widely used illicit drug by young people in high-income countries, and has recently become popular on a global scale. Epidemiological research during the past 10 years suggests that regular use of cannabis during adolescence and into adulthood can have adverse effects. Epidemiological, clinical, and laboratory studies have established an association between cannabis use and adverse outcomes. We focus on adverse health effects of greatest potential public health interest-that is, those that are most likely to occur and to affect a large number of cannabis users. The most probable adverse effects include a dependence syndrome, increased risk of motor vehicle crashes, impaired respiratory function, cardiovascular disease, and adverse effects of regular use on adolescent psychosocial development and mental health.

The development- and phencyclidine-regulated induction of synapse-associated protein-97 gene in the rat neocortex.

Using the RNA arbitrarily-primed PCR and the competitive RT-PCR, we have isolated the neocortical transcripts that are upregulated and unchanged in the adult and infant rats, respectively, after a systemic injection of an N-methyl-d-aspartate (NMDA) receptor antagonist phencyclidine (PCP), and found them identical to the synapse-associated protein-97 (SAP97) gene mRNAs. The upregulation of the SAP97 transcripts in the adult neocortex after the acute PCP injection was mimicked by another NMDA antagonist, dizocilpine, but not by the indirect dopamine agonists, methamphetamine and cocaine, a selective D1 receptor antagonist SCH23390, a D2 receptor-preferring antagonist haloperidol and a GABAergic anesthetic pentobarbital. Moreover, the pretreatment with a typical antipsychotic haloperidol failed to antagonize the increased neocortical SAP97 gene expression by PCP. These findings suggest that SAP97 might be involved in the molecular basis of the development-dependent onset of the non-dopaminergic symptoms seen in schizophrenia and the schizophrenia-like psychosis induced by NMDA receptor blocking.

Is ecstasy a drug of dependence

This paper examines the evidence for an MDMA or ecstasy dependence syndrome. Animal evidence suggests that MDMA may be a less potent reinforcer than other drugs, but that it does have dependence potential. This suggests that (a) ecstasy dependence might be less likely than dependence upon other drugs and (b) factors related to the behavioural and psychological aspects of reward and dependence may make a relatively greater contribution for ecstasy than for other drugs, where physically centred (and better understood) features of dependence may be more salient. Human evidence supports this proposition. Some people report problems with their use, but the literature suggests that physical features play a more limited role than psychological ones. Tolerance is apparent, and withdrawal is self-reported, but it is unclear whether these reports distinguish sub-acute effects of ecstasy intoxication from symptoms reflective of neuroadaptive processes underlying a true withdrawal syndrome. Studies examining the structure of dependence upon ecstasy suggest it may be different from drugs such as alcohol, methamphetamine and opioids. Consistent with studies of hallucinogens, a two-factor structure has been identified with factors suggestive of compulsive use and escalating use. Regardless of the nature of any dependence syndrome, however, there is evidence to suggest that a minority of ecstasy users become concerned about their use and seek treatment. Further controlled studies are required to investigate this phenomenon.

Prenatal exposure to phencyclidine produces abnormal behaviour and NMDA receptor expression in postpubertal mice.

Several studies have shown the disruptive effects of non-competitive N-methyl-d-aspartate (NMDA) receptor antagonists on neurobehavioural development. Based on the neurodevelopment hypothesis of schizophrenia, there is growing interest in animal models treated with NMDA antagonists at developing stages to investigate the pathogenesis of psychological disturbances in humans. Previous studies have reported that perinatal treatment with phencyclidine (PCP) impairs the development of neuronal systems and induces schizophrenia-like behaviour. However, the adverse effects of prenatal exposure to PCP on behaviour and the function of NMDA receptors are not well understood. This study investigated the long-term effects of prenatal exposure to PCP in mice. The prenatal PCP-treated mice showed hypersensitivity to a low dose of PCP in locomotor activity and impairment of recognition memory in the novel object recognition test at age 7 wk. Meanwhile, the prenatal exposure reduced the phosphorylation of NR1, although it increased the expression of NR1 itself. Furthermore, these behavioural changes were attenuated by atypical antipsychotic treatment. Taken together, prenatal exposure to PCP produced long-lasting behavioural deficits, accompanied by the abnormal expression and dysfunction of NMDA receptors in postpubertal mice. It is worth investigating the influences of disrupted NMDA receptors during the prenatal period on behaviour in later life.

Delta9-tetrahydrocannabinol (THC), 11-hydroxy-THC, and 11-nor-9-carboxy-THC plasma pharmacokinetics during and after continuous high-dose oral THC.

Delta(9)-tetrahydrocannabinol (THC) is the primary psychoactive constituent of cannabis and an active cannabinoid pharmacotherapy component. No plasma pharmacokinetic data after repeated oral THC administration are available. Six adult male daily cannabis smokers resided on a closed clinical research unit. Oral THC capsules (20 mg) were administered every 4-8 h in escalating total daily doses (40-120 mg) for 7 days. Free and glucuronidated plasma THC, 11-hydroxy-THC (11-OH-THC), and 11-nor-9-carboxy-THC (THCCOOH) were quantified by 2-dimensional GC-MS during and after dosing. Free plasma THC, 11-OH-THC, and THCCOOH concentrations 19.5 h after admission (before controlled oral THC dosing) were mean 4.3 (SE 1.1), 1.3 (0.5), and 34.0 (8.4) microgL, respectively. During oral dosing, free 11-OH-THC and THCCOOH increased steadily, whereas THC did not. Mean peak plasma free THC, 11-OH-THC, and THCCOOH concentrations were 3.8 (0.5), 3.0 (0.7), and 196.9 (39.9) mugL, respectively, 22.5 h after the last dose. Escherichia coli beta-glucuronidase hydrolysis of 264 cannabinoid specimens yielded statistically significant increases in THC, 11-OH-THC, and THCCOOH concentrations (P < 0.001), but conjugated concentrations were underestimated owing to incomplete enzymatic hydrolysis. Plasma THC concentrations remained >1 mugL for at least 1 day after daily cannabis smoking and also after cessation of multiple oral THC doses. We report for the first time free plasma THC concentrations after multiple high-dose oral THC throughout the day and night, and after Escherichia coli beta-glucuronidase hydrolysis. These data will aid in the interpretation of plasma THC concentrations after multiple oral doses.

A cannabinoid CB(1) receptor antagonist ameliorates impairment of recognition memory on withdrawal from MDMA (Ecstasy).

(-)-3,4-Methylenedioxymethamphetamine (MDMA, Ecstasy) abusers have persistent neuropsychiatric deficits including memory impairments after the cessation of abuse. On the other hand, cannabinoid CB(1) receptors have been implicated in learningmemory, and are highly expressed in the hippocampus, a region of the brain believed to have an important function in certain forms of learning and memory. In this study, we clarified the mechanism underlying the cognitive impairment that develops during MDMA withdrawal from the standpoint of the cannabinoid CB(1) receptors. Mice were administered MDMA (10 mgkg, i.p.) once a day for 7 days. On the 7th day of withdrawal, a novel object recognition task was performed and the amount of cannabinoid CB(1) receptor protein was measured with western blotting. Recognition performance was impaired on the 7th day of withdrawal. This impairment was blocked by AM251, a cannabinoid CB(1) receptor antagonist, administered 30 min before the training trial or co-administered with MDMA. At this time, the level of cannabinoid CB(1) receptor protein increased significantly in the hippocampus but not the prefrontal cortex or striatum. This increase of CB(1) receptor protein in the hippocampus was also blocked by the co-administration of AM251. Furthermore, CB(1) receptor knockout mice showed no impairment of recognition performance on the withdrawal from MDMA. The impairment of recognition memory during withdrawal from MDMA may result from the activation of cannabinoid CB(1) receptors in the hippocampus.

The psychotomimetic effects of short-term sensory deprivation.

People experiencing sensory deprivation often report perceptual disturbances such as hallucinations, especially over extended periods of time. However, there is little evidence concerning short-term sensory deprivation and whether its effects differ depending on the individual concerned, and in particular their proneness to psychosis. This study explored whether perceptual disturbances could be elicited by a brief period of complete isolation from sound and vision in both highly hallucination prone and nonhallucination prone groups. Greater psychotomimetic experiences taking the form of perceptual disturbances, paranoia, and anhedonia were found across both groups when under sensory deprivation. In addition, hallucination-prone individuals experienced more perceptual disturbances when placed in short-term sensory deprivation than nonprone individuals. This result is discussed in terms of difficulties in source monitoring as a possible mechanism involved in proneness to hallucinations.

A fatal case of myocardial damage due to misuse of the designer drug MDMA.

A 39-year-old woman collapsed after oral intake of 3,4-methylenedioxymethyl-amphetamine (MDMA, ecstasy). After ingestion of the drug, she had felt persistent discomfort in her anterior chest area, and lost consciousness for a few minutes on the following morning. She was transported to a hospital and died seven days after collapse. A serum sample obtained on admission revealed an MDMA concentration of 1.2mgL, but no evidence of any other drug including amphetamine, methamphetamine, or other ring-substituted amphetamines. Microscopic examination at autopsy revealed striking changes in the heart, including small foci of myocyte necrosis with a surrounding macrophage inflammatory response, foci of fibrosis, and calcification accompanied by myocyte necrosis, these changes being predominant in the right ventricle. In the liver, hepatic necrosis was observed with fatty degeneration accompanied by inflammation. Myoglobinuria was demonstrated in the kidney by immunohistochemistry. Degeneration of neurons throughout the whole brain was also evident, in addition to haemorrhagic foci in the pons and medulla. Serious bronchopneumonia was also found in the right lung. These findings provide evidence that oral intake of MDMA can result in cardiotoxicity, inducing cardiac arrhythmia and cardiovascular collapse. As a consequence of the compromised blood supply, brain necrosis may occur, followed by severe bronchopneumonia. Ingestion of MDMA could also lead to liver damage as well as myoglobinuria resulting from rhabdomyolysis. These data suggest that death in this case had been caused largely by MDMA intoxication.

MDMA pretreatment leads to mild chronic unpredictable stress-induced impairments in spatial learning.

3,4-Methylenedioxymethamphetamine (MDMA) is a drug of abuse worldwide and a selective serotonin (5-HT) neurotoxin. An important factor in the risk of drug abuse and relapse is stress. Although multiple parallels exist between MDMA abuse and stress, including effects on 5-HTergic neurotransmission, few studies have investigated the consequences of combined exposure to MDMA and chronic stress. Therefore, rats were pretreated with MDMA and exposed 7 days later to 10 days of mild chronic unpredictable stress (CUS). MDMA pretreatment was hypothesized to enhance the effects of CUS leading to enhanced 5-HT transporter (SERT) depletion in the hippocampus and increased anxiety and cognitive impairment. Whereas MDMA alone increased anxiety-like behavior on the elevated plus maze, CUS alone or in combination with MDMA pretreatment did not increase anxiety-like behavior. In contrast, MDMA pretreatment led to CUS-induced learning impairment in the Morris water maze but not an enhanced depletion of hippocampal SERT protein. These results show that prior exposure to MDMA leads to stress-induced impairments in learning behavior that is not otherwise observed with stress alone and appear unrelated to an enhanced depletion of SERT.

Electrospray MS-based characterization of beta-carbolines--mutagenic constituents of thermally processed meat.

The beta-carbolines 1-methyl-9H-pyrido 3,4-bindole and 9H-pyrido3,4bindole have been implicated as having causative roles in a number of human diseases, such as Parkinsons disease and cancer. As they can be formed during the heating of protein-rich food, a number of analytical methodologies have been proposed for their detection and quantification in foodstuff. For this purpose, LC-MS and LC-MSMS have emerged as the most specific analytical methods, and the quantification is based on the occurrence of unusual ions, such as MH-(H()) and MH-2H. In this study, we have investigated the formation of these ions by accurate-mass electrospray MSMS and demonstrated that these ions are formed from gas-phase ion-molecule reactions between water vapor present in the collision cell and the protonated molecule of 1-methyl-9H-pyrido 3,4-bindole and 9H-pyrido3,4bindole. Although this reaction has been previously described for heterocyclic amine ions, it has been overlooked in the most of recent LC-MS and LC-MSMS studies, and no complete data of the fragmentation are reported. Our results demonstrate that additional attention should be given with respect to eliminating water vapor residues in the mass spectrometer when analysis of beta-carbolines is performed, as this residue may affect the reliability in the results of quantification.

Long-term neuropsychological effects of ecstasy in middle-aged ecstasypolydrug users.

Studies reporting ecstasy-induced serotonin-toxicity and (neuro)psychological dysfunctions have been conducted in young adults. Little is known about ecstasy effects later in life, when serotonin levels and cognition decrease as a consequence of normal ageing. This study aimed to assess whether harmful effects of ecstasy only add to or also interact with age-related neuropsychological decline. Attention, verbal and visual memory, visuospatial ability, self-reported depression, sensation-seeking and impulsivity were assessed in middle-aged moderate to heavy ecstasypolydrug users (n 17) and compared with none or very mild ecstasy using polydrug users (matched for age, gender, intelligence and other drugs n 16) and a group of drug-naive controls (n 20). Moderate to heavy ecstasypolydrug users performed significantly worse on a verbal memory task than none or very mild ecstasy using polydrug users and drug naives. Moderate and heavy ecstasypolydrug users also differed significantly from drug-naives on measures of depression, sensation-seeking and impulsivity but not from none or very mild ecstasy-using polydrug users. This study in middle-aged ecstasypolydrug users replicated findings of studies in younger ecstasy users, showing a harmful effect of ecstasy on verbal memory. There was no clear support for an interaction between harmful effects of ecstasy use and age-related memory decline or mid-life depression.

Positive modulation of alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptors reverses sub-chronic PCP-induced deficits in the novel object recognition task in rats.

Cognitive deficits are a major clinical unmet need in schizophrenia. The psychotomimetic drug phencyclidine (PCP) is widely applied in rodents to mimic symptoms of schizophrenia, including cognitive deficits. Previous studies have shown that sub-chronic PCP induces an enduring episodic memory deficit in female Lister Hooded rats in the novel object recognition (NOR) task. Here we show that positive modulation of AMPA receptor (AMPAR) mediated glutamate transmission alleviates cognitive deficits induced by sub-chronic PCP treatment. Female Lister hooded rats were treated sub-chronically with either vehicle (0.9% saline) or PCP (2mgkg two doses per day for 7 days), followed by a 7 days washout period. 30 min prior to the acquisition trial of the NOR task animals were dosed with either vehicle, CX546 (10, 40 or 80 mgkg) or CX516 (0.5, 2.5, 10, 40 or 80 mgkg). Our results show that sub-chronic PCP treatment induced a significant decrease in the discrimination index (DI) and both ampakines CX546 and CX516 were able to reverse this disruption of object memory in rats in the novel object recognition task. These data suggest that positive AMPAR modulation may represent a mechanism for treatment of cognitive deficits in schizophrenia.

Cannabinoids for Tourettes Syndrome.

Gilles de la Tourette Syndrome (GTS) is a developmental neuropsychiatric disorder characterised by the presence of chronic motor and phonic tics. Drugs currently used in the treatment of GTS either lack efficacy or are associated with intolerable side effects. There is some anecdotal and experimental evidence that cannabinoids may be effective in treating tics and compulsive behaviour in patients with GTS. There are currently no systematic Cochrane reviews of treatments used in GTS. There is one other Cochrane review being undertaken at present, on the use of fluoxetine for tics in GTS. To evaluate the efficacy and safety of cannabinoids as compared to placebo or other drugs in treating tics, premonitory urges and obsessive compulsive symptoms (OCS), in patients with GTS. We searched the Cochrane Central Register of Controlled Trials (CENTRAL) (in The Cochrane Library Issue 4 2008) , MEDLINE (January 1996 to date), EMBASE (January 1974 to date), PsycINFO (January 1887 to date), CINAHL (January 1982 to date), AMED (January 1985 to date), British Nursing Index (January 1994 to date) and DH DATA (January 1994 to date). We also searched the reference lists of located trials and review articles for further information. We included randomised controlled trials (RCTs) comparing any cannabinoid preparation with placebo or other drugs used in the treatment of tics and OCS in patients with GTS. Two authors abstracted data independently and settled any differences by discussion. Only two trials were found that met the inclusion criteria. Both compared a cannabinoid, delta-9-Tetrahydrocannabinol (Delta(9)THC), either as monotherapy or as adjuvant therapy, with placebo. One was a double blind, single dose crossover trial and the other was a double blind, parallel group study. A total of 28 different patients were studied. Although both trials reported a positive effect from Delta(9)THC, the improvements in tic frequency and severity were small and were only detected by some of the outcome measures. Not enough evidence to support the use of cannabinoids in treating tics and obsessive compulsive behaviour in people with Tourettes syndrome.

Pain management and substance abuse a national dilemma.

Pain is the most common reason for medical care in the United States (Vukmir, 2004), with 32.8% of the U.S. population experiencing persistent or chronic pain symptoms. Undertreatment of pain is pervasive, as Congress declared 2001 to 2010 the decade of pain control in research and pharmaceutical companies responded with more effective medications to fight pain. However, nonmedical use and abuse of prescription pain medication has risen at an alarming rate in the United States within the past 5 years, especially among adolescents. The number of those abusing prescription drugs exceeds even combined numbers of those abusing cocaine, heroin, inhalants, and hallucinogens. This article examines the intersection of policies addressing nonmedical use and abuse of prescription pain medications and effects of enforcement strategies and policy direction on pain patients. Alternative policy direction and enforcement strategies are discussed.

Delta(9)-tetrahydrocannabinol regulates the p53 post-translational modifiers Murine double minute 2 and the Small Ubiquitin MOdifier protein in the rat brain.

The phytocannabinoid Delta(9)-Tetrahydrocannabinol (Delta(9)-THC), the main psychoactive cannabinoid in cannabis, activates a number of signalling cascades including p53. This study examines the role of Delta(9)-THC in regulating the p53 post-translational modifier proteins, Murine double minute (Mdm2) and Small Ubquitin-like MOdifier protein 1 (SUMO-1) in cortical neurons. Delta(9)-THC increased both Mdm2 and SUMO-1 protein expression and induced the deSUMOylation of p53 in a cannabinoid receptor type 1 (CB(1))-receptor dependent manner. We demonstrate that Delta(9)-THC decreased the SUMOylation of the CB(1) receptor. The data reveal a novel role for cannabinoid receptor activation in modulating the SUMO regulatory system.

Hollow fibre-supported liquid membrane extraction and LC-MSMS detection for the analysis of heterocyclic amines in urine samples.

Heterocyclic amines (HCAs) are potent mutagenscarcinogens to which humans are frequently exposed through the consumption of cooked meat and fish food. The effect of normal intake of HCAs and their role in the aetiology of human cancer is unknown. To some extent, limitations of the existing analytical methods in monitoring the low levels of HCAs in biological samples have hindered obtaining conclusive results. In this study, a method for the analysis of HCAs in human urine has been studied to detect HCAs and metabolites at levels resulting from consumption of food cooked at ordinary conditions. The analytical method consisted of extraction and clean-up by the novel technique liquid-phase microextraction combined with LC-MSMS. The effect of pH during the extraction and hydrolysis step was examined. High sensitivity was achieved when the extraction was performed in raw urine adjusted to pH 5.5, 2-amino-1-methyl-6-phenylimidazo4,5-bpyridine being detected from 2 pgg urine, levels comparable with a normal exposure. Good reproducibility and repeatability was obtained for 2-amino-1-methyl-6-phenylimidazo4,5-bpyridine and 2-amino-3,8-dimethylimidazo4,5-fquinoxaline, below 9% using isotopic dilution. The performance of the method on 9H-pyrido3,4-bindole, 2-amino-1-methyl-6-(4-hydroxyphenyl)imidazo4,5-bpyridine and 2-amino-1-methyl-6-(5-hydroxy)phenylimidazo4,5-bpyridine was also studied.

Randomized placebo-controlled double-blind clinical trial of cannabis-based medicinal product (Sativex) in painful diabetic neuropathy depression is a major confounding factor.

To assess the efficacy of Sativex, a cannabis-based medicinal extract, as adjuvant treatment in painful diabetic peripheral neuropathy (DPN). In this randomized controlled trial, 30 subjects with painful DPN received daily Sativex or placebo. The primary outcome measure was change in mean daily pain scores, and secondary outcome measures included quality-of-life assessments. There was significant improvement in pain scores in both groups, but mean change between groups was not significant. There were no significant differences in secondary outcome measures. Patients with depression had significantly greater baseline pain scores that improved regardless of intervention. This first-ever trial assessing the efficacy of cannabis has shown it to be no more efficacious than placebo in painful DPN. Depression was a major confounder and may have important implications for future trials on painful DPN.

Do Delta9-tetrahydrocannabinol concentrations indicate recent use in chronic cannabis users

To quantify blood Delta(9)-tetrahydrocannabinol (THC) concentrations in chronic cannabis users over 7 days of continuous monitored abstinence. Twenty-five frequent, long-term cannabis users resided on a secure clinical research unit at the US National Institute on Drug Abuse under continuous medical surveillance to prevent cannabis self-administration. Whole blood cannabinoid concentrations were determined by two-dimensional gas chromatography-mass spectrometry. Nine chronic users (36%) had no measurable THC during 7 days of cannabis abstinence 16 had at least one positive THC > or 0.25 ngml, but not necessarily on the first day. On day 7, 6 full days after entering the unit, six participants still displayed detectable THC concentrations mean - standard deviation (SD), 0.3 - 0.7 ngml and all 25 had measurable carboxy-metabolite (6.2 - 8.8 ngml). The highest observed THC concentrations on admission (day 1) and day 7 were 7.0 and 3.0 ngml, respectively. Interestingly, five participants, all female, had THC-positive whole blood specimens over all 7 days. Body mass index did not correlate with time until the last THC-positive specimen (n 16 r -0.2 P 0.445). Substantial whole blood THC concentrations persist multiple days after drug discontinuation in heavy chronic cannabis users. It is currently unknown whether neurocognitive impairment occurs with low blood THC concentrations, and whether return to normal performance, as documented previously following extended cannabis abstinence, is accompanied by the removal of residual THC in brain. These findings also may impact on the implementation of per se limits in driving under the influence of drugs legislation.

Content of ecstasy in the Netherlands 1993-2008.

The present paper outlines the results of analyses carried out on the content of tablets sold as ecstasy, collected in the Netherlands by the Drugs Information Monitoring System (DIMS) from January 1993 to December 2008. During a period of 16 years, the DIMS analysed the content of 33 006 tablets sold as ecstasy that were handed in by numerous individual (potential) substance users. The DIMS results were compared with the results from various seized tablets to determine whether the DIMS is a monitor of the ecstasy consumer market. The DIMS system appears to be a market monitor that gives an accurate reflection of what is actually available on the hidden Dutch ecstasy market. During 16 years of monitoring, the purity tablets containing only 3,4-methylenedioxymethamphetamine (MDMA) was lowest around 1997. During this time-period many tablets contained other substances in addition to or instead of MDMA e.g. 3,4-methylene-dioxyamphetamine (MDA), 3,4-methylene-dioxyethylamphetamine (MDEA) and N-methyl-a-(1,3-benzodixol-5-yl)-2-butamine (MBDB), amphetamine and caffeine. From 1998 to 2008, the number of high-dose tablets (> or 106 mg MDMA per tablet) gradually increased. The same holds true for the proportion of tablets that contained only MDMA, reaching the highest levels in 2000 and 2004. After 2004, the purity of ecstasy tablets decreased again, caused mainly by a growing proportion of tablets containing meta-chlorophenylpiperazine (mCPP). The DIMS results provide valuable qualitative information on the content of ecstasy tablets in the Netherlands, and its changes throughout the years. Moreover, the results were used for national and international risk assessments and important warning and prevention activities.

Extending drug ethno-epidemiology using agent-based modelling.

To show how the inclusion of agent-based modelling improved the integration of ethno-epidemiological data in a study of psychostimulant use and related harms among young Australians. Agent-based modelling, ethnographic fieldwork, in-depth interviews and epidemiological surveys. Melbourne, Perth and Sydney, Australia. Club drug users in Melbourne, recreational drug users in Perth and street-based injecting drug users in Sydney. Participants were aged 18-30 years and reported monthly or more frequent psychostimulant use. Agent-based modelling provided a specific focus for structured discussion about integrating ethnographic and epidemiological methods and data. The modelling process was underpinned by collective and incremental design principles, and produced SimAmph, a data-driven model of social and environmental agents and the relationships between them. Using SimAmph, we were able to test the probable impact of ecstasy pill-testing on the prevalence of harms--a potentially important tool for policy development. The study also navigated a range of challenges, including the need to manage epistemological differences, changes in the collective design process and modelling focus, the differences between injecting and non-injecting samples and concerns over the dissemination of modelling outcomes. Agent-based modelling was used to integrate ethno-epidemiological data on psychostimulant use, and to test the probable impact of a specific intervention on the prevalence of drug-related harms. It also established a framework for collaboration between research disciplines that emphasizes the synthesis of diverse data types in order to generate new knowledge relevant to the reduction of drug-related harms.

The association between the negative effects attributed to ecstasy use and measures of cognition and mood among users.

In self reports, abstinent ecstasypolydrug users claim that they experience certain ongoing affective and psychological changes including elevated anxiety, arousal, and depression. In addition, various aspects of cognition (e.g., everyday memory, reasoning, executive functioning) appear to be affected. The present paper investigated the link between these two psychological sequelae. Ninety-five ecstasypolydrug users completed tests of reasoning, intelligence, information processing speed, executive functioning, and everyday memory. Affect was measured via a mood adjective checklist. Adverse effects attributed to ecstasy were measured via responses to adjectives reflecting changes in users since they started using the drug. In addition, indicators of sleep quality and daytime sleepiness were obtained. Users attributed a number of adverse effects to ecstasy, namely heightened irritability, depression, paranoia, and deteriorating health. Adverse effects were significantly and negatively correlated with aspects of intelligence, everyday memory, and sleep quality. Length of use of ecstasy use was positively correlated with adverse effects. While many users attribute a number of adverse affects to their use of ecstasy, it remains unclear whether these self-perceptions are a corollary of the psychopharmacological effects of the drug or reflect factors which in fact predate its use.

Emerging treatments for PTSD.

Recent innovations in posttraumatic stress disorder (PTSD) research have identified new treatments with significant potential, as well as novel enhancements to empirically-validated treatments. This paper reviews emerging psychotherapeutic and pharmacologic interventions for the treatment of PTSD. It examines the evidence for a range of interventions, from social and family-based treatments to technological-based treatments. It describes recent findings regarding novel pharmacologic approaches including propranolol, ketamine, prazosin, and methylenedioxymethamphetamine. Special emphasis is given to the description of virtual reality and D-cycloserine as enhancements to prolonged exposure therapy.

Mifepristone and spironolactone differently alter cocaine intravenous self-administration and cocaine-induced locomotion in C57BL6J mice.

Corticosterone, the main glucorticoid hormone in rodents, facilitates behavioral responses to cocaine. Corticosterone is proposed to modulate cocaine intravenous self-administration (SA) and cocaine-induced locomotion through distinct receptors, the glucocorticoid receptor (GR) and the mineralocorticoid receptor (MR), respectively. However, this remains debatable. On one hand, modulation of both responses by the GR was tested in different experimental conditions, i.e. light versus dark nycthemeral phase and naïve versus cocaine-experienced animals. On the other hand, modulation of both responses by the MR was never tested directly but only inferred based on the ability of low plasma corticosterone levels (those for which corticosterone almost exclusively binds the MR) to compensate the effects of adrenalectomy. Our goal here was to test the involvement of the GR and the MR in cocaine-induced locomotor and reinforcing effects in the same experimental conditions. C57Bl6J mice were trained for cocaine (1 mgkginfusion) intravenous SA over 40 SA sessions. The animals were then administered with mifepristone (30 mgkg i.p.), a GR antagonist, or with spironolactone (20 mgkgi.p.), an MR antagonist, 2 hours before either cocaine intravenous SA or cocaine-induced locomotion. In a comparable nycthemeral period and in similarly cocaine-experienced animals, a blockade of the GR decreased cocaine-induced reinforcing effects but not cocaine-induced locomotion. A blockade of the MR decreased both cocaine-induced reinforcing (but to a much lesser extent than the GR blockade) and locomotor effects. Altogether, our results comforted the hypothesis that the GR modulates cocaine-related operant conditioning, while the MR would modulate cocaine-related unconditioned effects. The present data also reveal mifepristone as an interesting tool for manipulating the impact of corticosterone on cocaine-induced reinforcing effects in mice.

Chemical interactions with pyramidal neurons in layer 5 of the cerebral cortex control of pain and anxiety.

Pyramidal neurons in layer 5 of the cerebral cortex are involved in learning and memory and have complex connections with other neurons through a very large array of dendrites. These dendrites can switch between long term depression and long term potentiation depending on global summation of various inputs. The plasticity of the input into pyramidal neurons makes the neuronal output variable. Many interneurons in the cerebral cortex and distant neurons in other brain regions are involved in providing input to pyramidal neurons. All of these neurons and interneurons have neurotransmitters that act through receptors to provide input to pyramidal neurons. Serotonin is one of the important neurotransmitters involved with pyramidal neurons and has been implicated in psychosis, psychedelic states and what are called sacred dreams. This review will discuss the various chemicals and receptors that are important with pyramidal neurons including opioids, nicotine, scopolamine, psilocybin, LSD, mescaline, ergot alkaloids, salvinorin A, ergine and other compounds that interact with opioid, nicotinic, muscarinic and serotonergic receptors. The natural compounds provide clues to structure activity relationships with the receptors. It has been postulated that each receptor in the body has a natural agonist and antagonist, in addition to the normal neurotransmitters. It is common for natural antagonists and agonists to be peptides. Various possible peptide structures will be proposed for natural antagonists and agonists at each receptor. Natural antagonists and agonists may provide new ways to explore the functions of pyramidal neurons in normal health and pain management.

Enzymatic-nonenzymatic cellular antioxidant defense systems response and immunohistochemical detection of MDMA, VMAT2, HSP70, and apoptosis as biomarkers for MDMA (Ecstasy) neurotoxicity.

3,4-Methylenedioxymethamphetamine (MDMA)-induced neurotoxicity leads to the formation of quinone metabolities and hydroxyl radicals and then to the production of reactive oxygen species (ROS). We evaluated the effect of a single dose of MDMA (20 mgkg, i.p.) on the enzymatic and nonenzymatic cellular antioxidant defense system in different areas of rat brain in the early hours (<6 hr) of the administration itself, and we identified the morphological expressions of neurotoxicity induced by MDMA on the vulnerable brain areas in the first 24 hr. The acute administration of MDMA produces a decrease of reduced and oxidized glutathione ratio, and antioxidant enzyme activities were significantly reduced after 3 hr and after 6 hr in frontal cortex. Ascorbic acid levels strongly increased in striatum, hippocampus, and frontal cortex after 3 and 6 hr. High levels of malonaldehyde with respect to control were measured in striatum after 3 and 6 hr and in hippocampus and frontal cortex after 6 hr. An immunohistochemical investigation on the frontal, thalamic, hypothalamic, and striatal areas was performed. A strong positive reaction to the antivesicular monoamine transporter 2 was observed in the frontal section, in the basal ganglia and thalamus. Cortical positivity, located in the most superficial layer was revealed only for heat shock protein 70 after 24 hr.

Simultaneous screening and quantification of 29 drugs of abuse in oral fluid by solid-phase extraction and ultraperformance LC-MSMS.

The European DRUID (Driving under the Influence of Drugs, Alcohol And Medicines) project calls for analysis of oral fluid (OF) samples, collected randomly and anonymously at the roadside from drivers in Denmark throughout 2008-2009. To analyze these samples we developed an ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MSMS) method for detection of 29 drugs and illicit compounds in OF. The drugs detected were opioids, amphetamines, cocaine, benzodiazepines, and Delta-9-tetrahydrocannabinol. Solid-phase extraction was performed with a Gilson ASPEC XL4 system equipped with Bond Elut Certify sample cartridges. OF samples (200 mg) diluted with 5 mL of ammonium acetatemethanol (volvol 9010) buffer were applied to the columns and eluted with 3 mL of acetonitrile with aqueous ammonium hydroxide. Target drugs were quantified by use of a Waters ACQUITY UPLC system coupled to a Waters Quattro Premier XE triple quadrupole (positive electrospray ionization mode, multiple reaction monitoring mode). Extraction recoveries were 36%-114% for all analytes, including Delta-9-tetrahydrocannabinol and benzoylecgonine. The lower limit of quantification was 0.5 mugkg for all analytes. Total imprecision (CV) was 5.9%-19.4%. With the use of deuterated internal standards for most compounds, the performance of the method was not influenced by matrix effects. A preliminary account of OF samples collected at the roadside showed the presence of amphetamine, cocaine, codeine, Delta-9-tetrahydrocannabinol, tramadol, and zopiclone. The UPLC-MSMS method makes it possible to detect all 29 analytes in 1 chromatographic run (15 min), including Delta-9-tetrahydrocannabinol and benzoylecgonine, which previously have been difficult to incorporate into multicomponent methods.

Methylenedioxymethamphetamine-related deaths in Taiwan 2001-2008.

Methylenedioxymethamphetamine (MDMA) has been one of most popular drugs in the club scene in Taiwan. This epidemic was studied through the examination of toxicological data obtained from the 59 fatalities tested positive for MDMA during the period of January 2001 to December 2008. Ketamine was found in 28 of these cases, signifying the popularity of this drug in Taiwan. The annual number of deaths in each of the 8 years in this period was 4, 7, 9, 14, 8, 9, 2, and 6, respectively. Among these 59 deaths, 39 (66.1%) were men, and the mean, median, and range of ages were 24.6, 23, and 14-46, respectively. Causes of death ruled by the attending pathologists and the distributions for these fatalities were acute intoxication, 40 and mechanical injury, 19, including 3 hanging and 2 drowning. The manners of death were ruled as accidental, 44 homicidal, 6 suicidal, 7 and undetermined, 2. In this study, postmortem whole blood was analyzed by gas chromatography-mass spectrometry with a limit of quantitation at 0.05 microgmL for both MDMA and MDA. The mean, median, and range of MDMA concentrations in the cases, where MDMA acute intoxication was ruled as the cause of death, were 4.75, 2.60, and 0.12-40.41 microgmL. MDA was found in 30 of these 40 cases with the following mean, median, and range data 0.19, 0.13, and 0.05-1.81 microgmL. The corresponding data of MDMA and MDA in the remaining 19 MDMA-related deaths were significantly lower 1.25, 0.97, 0.08-3.05 and 0.11, 0.09, 0.06-0.24 microgmL, respectively.

Comprehensive analysis of drugs of abuse in urine using disposable pipette extraction.

The extraction of basic, acidic, and neutral drugs of abuse from a low volume of urine (0.2 mL) using disposable pipette extraction (DPX) is described. DPX is a solid-phase extraction device that uses loosely contained sorbent inside a pipette tip fitted with a screen. This device provides faster extraction times because conditioning steps are not required. In this study, the DPX used a modified divinyl benzene sorbent containing both cation-exchange and reversed-phase mechanisms that facilitates the retention of basic and acidicneutral drugs, respectively. With this device, a comprehensive method of analysis was developed for a diverse group of drugs and drug classes in urine including amphetamines, opiates, cocaine and its metabolites, tetrahydrocannabinol metabolite, tricyclic antidepressants, meperidine, methadone, and phencyclidine. Recoveries of the majority of drugs analyzed were 90% or greater with relative standard deviations of less than 10%. Additional validation involved the analysis of urine specimens previously analyzed by a local forensic toxicology laboratory.

Harmine specifically inhibits protein kinase DYRK1A and interferes with neurite formation.

DYRK1A is a dual-specificity protein kinase that autophosphorylates a conserved tyrosine residue in the activation loop but phosphorylates exogenous substrates only at serine or threonine residues. Tyrosine autophosphorylation of DYRKs is a one-off event that takes place during translation and induces the activation of the kinase. Here we characterize the beta-carboline alkaloid harmine as a potent and specific inhibitor of DYRK1A both in vitro and in cultured cells. Comparative in vitro assays of four kinases of the DYRK family showed that harmine inhibited substrate phosphorylation by DYRK1A more potently than it inhibited substrate phosphorylation by the closely related kinase DYRK1B half maximal inhibitory concentrations (IC(50)) of 33 nm versus 166 nm, respectively and by the more distant members of the family, DYRK2 and DYRK4 (1.9 microm and 80 microm, respectively). Much higher concentrations of harmine were required to suppress tyrosine autophosphorylation of the translational intermediate of DYRK1A in a bacterial in vitro translation system (IC(50) 1.9 microm). Importantly, harmine inhibited the phosphorylation of a specific substrate by DYRK1A in cultured cells with a potency similar to that observed in vitro (IC(50) 48 nm), without negative effects on the viability of the cells. Overexpression of the DYRK1A gene on chromosome 21 has been implicated in the altered neuronal development observed in Down syndrome. Here, we show that harmine interferes with neuritogenesis in cultured hippocampal neurons. In summary, our data show that harmine inhibits DYRK1A substrate phosphorylation more potently than it inhibits tyrosine autophosphorylation, and provide evidence for a role of DYRK1A in the regulation of neurite formation.

Knowledge of Dentistry, Medicine, and Pharmacy Students about Psychedelic Drugs in Kerman University of Medical Sciences.

Psychedelic drugs can cause one to get out of normal status and permanent cerebral defects, via affecting central nervous system. Consumption of theses drugs seems to be increasing nowadays especially among the youth and university educated population. We conducted a study to evaluate the awareness of medical science students of Kerman University of medical science who are going to be the future medical population. This cross-sectional study was carried out on 471 of students of medicine, dentistry and pharmacy which were in the first to forth year of their education about psychedelic drugs (Ecstasy, LSD, Ice, crack and Yaba). To evaluate the students awareness of drugs we used questionnaire with reliability and validity proven via pilot study. Statistics analysis was performed using SPSS13 software. Average of their age was 3.2 ± 20.4. Overall among the students, 56.7% were in the low level of insight, 34.3% in medium and 6.9% in good level and 2.2% had best insight of the drugs. Also only 32.2% of students had the full information about the name of drug, 25.7 % had information about the form of them, 24% about the addiction with them, 7% about their complication and only 5% about the origin of drugs. The information about all psychedelic drugs was higher among pharmacy students, students of the third year and males. Our study showed a low insight about psychedelic drugs like Ecstasy, LSD, Ice, Crack, and Yaba among the students. According to this lack of information of these groups, it is suggested that educational courses about the complication, signs and symptoms of these drugs be held.

Amphetamine and its analogues determination in urine from patients hospitalized in the Department of Toxicology Jagiellonian University Collegium Medicum in Krakow.

The most popular stimulant drugs in Poland are amphetamine and its analogues ecstasy (MDMA) and methamphetamine (MA). The substances are usually determined by immunoassays (EMIT, FPIA, Rapid Tests) in medical, toxicological laboratories. The methods are not useful for identification of the substances. Chromatography methods are reference, confirmation methods that identify stimulant drugs in biological samples. The aim of the study was investigation of kinds of amphetamine derivatives abused by poisoned patients treated in the Clinic of Toxicology of the Jagiellonian University Collegium Medicum in Krakow. Materials for the study were urine samples collected from 46 patients. All the samples were positive for amphetamine presence by EMIT (cut off 300nn mL). Amphetamine analogues were identified by high performance liquid chromatography with diode array detector (HPLC-DAD). Amphetamine alone was present in 54.3% samples amphetamine and its analogues (MDMA, MDA, MA) were present in 26.0% MDA and MDMA (no amphetamine) were present in 19.5% samples. In one case no amphetamine neither its analogues were detected in the sample. The positive amphetamine EMIT result was caused by fenfluramine--the substance present in slimming product. the most often determined stimulant drug in the group of poisoned patients was amphetamine. The frequency of MDMA and MA intoxications were occasional and with probably increase. Immunoassay tests are useful for routine analysis in clinic laboratories but in some cases when the history is unreliable, confirmation analysis by reference method should be performed to identify the abused substance.

A behavioural comparison of acute and chronic Delta9-tetrahydrocannabinol and cannabidiol in C57BL6JArc mice.

Cannabis contains over 70 unique compounds and its abuse is linked to an increased risk of developing schizophrenia. The behavioural profiles of the psychotropic cannabis constituent Delta9-tetrahydrocannabinol (Delta9-THC) and the non-psychotomimetic constituent cannabidiol (CBD) were investigated with a battery of behavioural tests relevant to anxiety and positive, negative and cognitive symptoms of schizophrenia. Male adult C57BL6JArc mice were given 21 daily intraperitoneal injections of vehicle, Delta9-THC (0.3, 1, 3 or 10 mgkg) or CBD (1, 5, 10 or 50 mgkg). Delta9-THC produced the classic cannabinoid CB1 receptor-mediated tetrad of hypolocomotion, analgesia, catalepsy and hypothermia while CBD had modest hyperthermic effects. While sedative at this dose, Delta9-THC (10 mgkg) produced locomotor-independent anxiogenic effects in the open-field and light-dark tests. Chronic CBD produced moderate anxiolytic-like effects in the open-field test at 50 mgkg and in the light-dark test at a low dose (1 mgkg). Acute and chronic Delta9-THC (10 mgkg) decreased the startle response while CBD had no effect. Prepulse inhibition was increased by acute treatment with Delta9-THC (0.3, 3 and 10 mgkg) or CBD (1, 5 and 50 mgkg) and by chronic CBD (1 mgkg). Chronic CBD (50 mgkg) attenuated dexamphetamine (5 mgkg)-induced hyperlocomotion, suggesting an antipsychotic-like action for this cannabinoid. Chronic Delta9-THC decreased locomotor activity before and after dexamphetamine administration suggesting functional antagonism of the locomotor stimulant effect. These data provide the first evidence of anxiolytic- and antipsychotic-like effects of chronic but not acute CBD in C57BL6JArc mice, extending findings from acute studies in other inbred mouse strains and rats.

Recreational use of benzydamine as a hallucinogen among street youth in Brazil.

To describe the recreational use of benzydamine, an anti-inflammatory drug, among street youth in Brazil. a descriptive, cross sectional survey. 93 welfare services for the street youth in 27 Brazilian capitals. 2807 street youth, 10 to 18 years old. demographic characteristics, drug use pattern (lifetime use, use in the past 30 days, frequency, and characteristics of use in the past month) and effects of benzydamine through the use of a questionnaire. 78 reported lifetime recreational benzydamine use (67 cases identified only in three capitals). Among the 30 respondents reporting drug use in the last month (the month preceding the survey), 66.7% (n 20) used the drug on 4 or more days (in the month preceding the survey). The most frequently (50%) pleasure effects reported were hallucination and nonspecific sensory changes described as trips. Unwanted effects were reported by 75% of respondents, they were especially nausea and vomiting (21.4%). In the majority of the cases, drug was obtained from drugstores without a medical prescription. This study identifies the recreational use of benzydamine among street youth, mainly in the Northeast of Brazil, and also indicates the need for special controls on the dispensation of this substance.

Methamphetamine- and 3,4-methylenedioxymethamphetamine-induced behavioral changes in histamine H3-receptor knockout mice.

Histamine H(3) receptors inhibit the release of not only histamine itself, but also other neurotransmitters including dopamine. Previous papers have reported that histaminergic neurons inhibit psychostimulant-induced behavioral changes. To examine whether deficiency in histamine H(3) receptors influences psychostimulant-induced behavioral sensitization and reward, we examined locomotor activity, conditioned place preference (CPP), and c-Fos expression in histamine H(3) receptor-gene knockout mice (H3KO) and their wild-type (WT) counterparts before and after treatment with methamphetamine (METH) and 3,4-methylenedioxymethamphetamine (MDMA). The increase in locomotion induced by treatment with METH or MDMA was lower in histamine H3KO mice than in WT mice, while the locomotor sensitization was developed by METH or MDMA in both strains. However, no significant difference in METH- and MDMA-induced preference scores of CPP between histamine H3KO mice and WT mice was observed. Following treatment with METH, the number of c-Fos-positive neurons in the the caudate-putamen of histamine H3KO mice was lower than that in the caudate-putamen of WT mice. In contrast, there was no significant difference in the number of the psychostimulant-induced c-Fos-positive cells in the nucleus accumbens between the two strains of mice. These findings suggest that deficiency in histamine H(3) receptors may have inhibitory effects on psychostimulant-induced increase in locomotion, but insignificant effects on the reward.

Daily marijuana users with past alcohol problems increase alcohol consumption during marijuana abstinence.

Drug abuse treatment programs typically recommend complete abstinence because of a fear that clients who stop use of one drug will substitute another. A within-subjects study investigated whether consumption of alcohol and other substances changes during marijuana abstinence. Twenty-eight daily marijuana users who were not trying to stop or reduce their marijuana consumption completed an 8-day baseline period in which they used marijuana and other drugs as usual, a 13-day marijuana abstinence period, and a 7-day return-to-baseline period. Participants provided self-report of substance use daily and submitted urine samples twice weekly to verify marijuana abstinence. A diagnosis of past alcohol abuse or dependence significantly moderated the alcohol increase from baseline to marijuana abstinence (p<0.01), such that individuals with this diagnosis significantly increased alcohol use (52% increase) but those without this history did not (3% increase). Increases in marijuana withdrawal discomfort scores and alcohol craving scores from baseline to marijuana abstinence significantly and positively correlated with increases in alcohol use. Increases in cigarettes, caffeine, and non-marijuana illicit drugs did not occur. This study provides empirical validation of drug substitution in a subgroup of daily marijuana users, but results need to be replicated in individuals who seek treatment for marijuana problems.