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In the last decade, the market for new psychoactive substances has been enriched by numerous psychedelic phenethylamines, which mimic the psychoactive effect of lysergic acid diethylamide (LSD). In particular, the -NBOMe series, which are more potent than their 2C compounds analogs, are considered worthy substitutes for LSD by users. The purpose of this study was to assess the effects of 25

Fentanyl derivatives (FENS) belongs to the class of Novel Synthetic Opioids that emerged in the illegal drug market of New Psychoactive Substances (NPS). These substances have been implicated in many cases of intoxication and death with overdose worldwide. Therefore, the aim of this study is to investigate the pharmaco-dynamic profiles of three fentanyl (FENT) analogues: Acrylfentanyl (ACRYLF), Ocfentanyl (OCF) and Furanylfentanyl (FUF). In vitro, we measured FENS opioid receptor efficacy, potency, and selectivity in calcium mobilization studies performed in cells coexpressing opioid receptors and chimeric G proteins and their capability to promote the interaction of the mu receptor with G protein and β-arrestin 2 in bioluminescence resonance energy transfer (BRET) studies. In vivo, we investigated the acute effects of the systemic administration of ACRYLF, OCF and FUF (0.01-15 mg/kg i.p.) on mechanical and thermal analgesia, motor impairment, grip strength and cardiorespiratory changes in CD-1 male mice. Opioid receptor specificity was investigated in vivo using naloxone (NLX; 6 mg/kg i.p) pre-treatment. In vitro, the three FENS were able to activate the mu opioid receptor in a concentration dependent manner with following rank order potency: FUF FENT=OCF ACRYLF. All compounds were able to elicit maximal effects similar to that of dermorphin, with the exception of FUF which displayed lower maximal effects thus behaving as a partial agonist. In the BRET G-protein assay, all compounds behaved as partial agonists for the β-arrestin 2 pathway in comparison with dermorphin, whereas FUF did not promote β-arrestin 2 recruitment, behaving as an antagonist. In vivo, all the compounds increased mechanical and thermal analgesia with following rank order potency ACRYLF = FENT FUF OCF and impaired motor and cardiorespiratory parameters. Among the substances tested, FUF showed lower potency for cardiorespiratory and motor effects. These findings reveal the risks associated with the use of FENS and the importance of studying the pharmaco-dynamic properties of these drugs to better understand possible therapeutic interventions in the case of toxicity.

N-methyl-2-pyrrolidone (NMP) and γ-hydroxybutyrate acid (GHB) are synthetic solvents detected in the recreational drug market. GHB has sedative/hypnotic properties and is used for criminal purposes to compromise reaction ability and commit drug-facilitated sexual assaults and other crimes. NMP is a strong solubilizing solvent that has been used alone or mixed with GHB in case of abuse and robberies. The aim of this experimental study is to compare the acute pharmaco-toxicological effects of NMP and GHB on neurological signs (myoclonia, convulsions), sensorimotor (visual, acoustic, and overall tactile) responses, righting reflex, thermoregulation, and motor activity (bar, drag, and accelerod test) in CD-1 male mice. Moreover, since cardiorespiratory depression is one of the main adverse effects related to GHB intake, we investigated the effect of NMP and GHB on cardiorespiratory changes (heart rate, breath rate, oxygen saturation, and pulse distension) in mice. The present study demonstrates that NMP inhibited sensorimotor and motor responses and induced cardiorespiratory depression, with a lower potency and efficacy compared to GHB. These results suggest that NMP can hardly be used alone as a substance to perpetrate sexual assault or robberies.

Introduction: The anterior cingulate cortex (ACC) has shown to play a role in impulsivity, fear, and anxiety. Considering, its high glutamate receptor density, it was chosen as a region of interest to investigate the role of glutamate transmission in drug dependance. We investigated the correlations between personality trait scores and glutamate-to-glutamine (Glx) ratio concentrations in the ACC in order to evaluate if (1) personality traits may increase the probability of drug use and (2) drug use can modify cerebral metabolic pattern contributing to addictive behaviors. Materials and Methods: Glx ratio concentrations in the ACC region were measured with high-resolution multivoxel proton magnetic resonance spectroscopy (1H-MRS). Personality traits were evaluated utilizing Cloninger's TCI-revised test. Bivariate correlations between personality scores of 28 teens cannabis users (males, mean age = 18.54 ± 2.80) were evaluated. Results: In the ACC, we observed negative correlation between GG concentrations (r = −0.44, P = 0.05) and co-operativeness values (CO), choline (cho), and novelty seeking (NS) values (r = −0,45, P = 0.05). Low levels of glutamate and high levels of cho in the ACC were closely related to the CO and NS personality traits. Conclusions: Metabolic and personality patterns seems to be related to the risk of substance predisposition in adolescents. Our data contribute a possible support to the “top-down” control of the ACC on brain metabolism, due to the particular cerebral metabolic pattern found in “drug-using” adolescents.

MT-45 is a synthetic opioid that was developed in the 1970s as an analgesic compound. However, in recent years MT-45 has been associated with multiple deaths in Europe and has been included in the class of novel psychoactive substances known as novel synthetic opioids (NSOs). Little is known about the pharmaco-toxicological effects of MT-45. Therefore, we used a dynamic mass redistribution (DMR) assay to investigate the pharmacodynamic profile of this NSO in vitro compared with morphine. We then used in vivo studies to investigate the effect of the acute systemic administration of MT-45 (0.01–15 mg/kg i.p.) on motor and sensorimotor (visual, acoustic and tactile) responses, mechanical and thermal analgesia, muscle strength and body temperature in CD-1 male mice. Higher doses of MT-45 (6–30 mg/kg i.p.) were used to investigate cardiorespiratory changes (heart rate, respiratory rate, SpO2 saturation and pulse distention). All effects of MT-45 were compared with those of morphine. In vitro DMR assay results demonstrated that at human recombinant opioid receptors MT-45 behaves as a potent selective mu agonist with a slightly higher efficacy than morphine. In vivo results showed that MT-45 progressively induces tail elevation at the lowest dose tested (0.01 mg/kg), increased mechanical and thermal antinociception (starting from 1 to 6 mg/kg), decreased visual sensorimotor responses (starting from 3 to 6 mg/kg) and reduced tactile responses, modulated motor performance and induced muscle rigidity at higher doses (15 mg/kg). In addition, at higher doses (15–30 mg/kg) MT-45 impaired the cardiorespiratory functions. All effects were prevented by the administration of the opioid receptor antagonist naloxone. These findings reveal the risks associated with the ingestion of opioids and the importance of studying these drugs and undertaking more clinical studies of the current molecules to better understand possible therapeutic interventions in the case of toxicity.

Background Reports concerning the causal link between aggressive behaviour and use and abuse of different substances (i.e., alcohol, MDPV) can be found in the literature. Nonetheless, the topic concerning the effects of acute ethanol administration on MDPV and cocaine induced aggressive behaviour has yet to be thoroughly investigated. The aim of this study was to investigate such synergistic effects. Materials and methods A total of 360 male mice were employed in the study. Ethanol was diluted with saline solution and administered 10 minutes before MDPV or cocaine injection via oral gavage needles. Similarly, MDPV and cocaine were dissolved in saline solution and administered by intraperitoneal injection. Different associations of specific drug doses were then tested. To investigate the acute effects of MDPV and cocaine and their interaction with ethanol on aggression in mice, a resident–intruder test was used. Results Ethanol alone was ineffective at dosages of 0.05 g/kg and 0.25 g/kg but increased the aggressiveness of the mice at 0.125 g/kg. Similarly, the injection of both cocaine alone and MDPV alone did not significantly increase the aggressiveness of the mice; conversely, the combination of ethanol and cocaine and ethanol and MDPV enhanced aggression at specific ethanol dosages (0.05 g/kg and 0.125 g/kg). Conclusion This study demonstrated that acute ethanol administration enhances MDPV- and cocaine-induced aggressive behaviour in mice. This aggressive response is particularly enhanced when MDVP and cocaine are coupled with specific ethanol dosages, proving that psychostimulant drugs may act synergistically under certain conditions.

MDPV is a synthetic cathinone illegally marketed and consumed for its psychostimulant effects, which are similar to those produced by cocaine, amphetamines, and MDMA. Clinical reports indicate that MDPV produces euphoria, increases alertness, and at high doses causes agitation, psychosis, tachycardia and hypertension, hallucinations, delirium, hyperthermia, rhabdomyolysis, and even death. In rodents, MDPV reproduces the typical physiological effects of psychostimulant drugs, demonstrating greater potency than cocaine. Nevertheless, its role in aggressive behavior has been reported but not yet experimentally confirmed. Therefore, the aim of this study was to evaluate the effects of acute and repeated MDPV (0.01-10 mg/kg i.p.) administration on aggressive behavior in mice and to compare them with those of cocaine (0.01-10 mg/kg i.p.) administration. To this purpose, the resident-intruder test in isolated mice and the spontaneous and stimulated aggressiveness tests for group-housed mice were employed. The present study shows for the first time that MDPV enhances aggressive behavior and locomotion in mice with greater potency and efficacy than cocaine treatment. Moreover, the aggressive and locomotor responses are enhanced after repeated administration, indicating that a sensitization mechanism comes into play. These results, although from preclinical investigation, are suggestive that human MDPV intake could be a problem for public health and the criminal justice system. Thus, investigation by police officers and medical staff is needed to prevent interpersonal violence induced by the consumption of synthetic cathinones.

JWH-018-Cl, JWH-018-Br and AM-2201 (JWH-018 halogenated-derivatives; JWH-018-R compounds) are synthetic cannabinoid agonists illegally marketed as “Spice”, “K2”, “herbal blend” and research chemicals for their cannabis-like psychoactive effects. In rodents, JWH-018 and its halogenated derivatives reproduce the typical effects of Δ9-tetrahydrocannabinol (Δ9-THC), i.e. hypothermia, analgesia, hypolocomotion and akinesia. Yet, the effects of JWH-018-R compounds on sensorimotor functions are still unknown. This study was designed to investigate the effect of an acute intraperitoneal (i.p.) administration of JWH-018-R compounds (0.01–6 mg/kg) on sensorimotor functions in mice and to compare them to those caused by the reference compound JWH-018 and Δ9-THC. A well validated battery of behavioral tests was used to investigate the effects of these synthetic cannabinoids on the visual, auditory and tactile responses in mice, while the pre-pulse inhibition (PPI) test was used to investigate their effect on sensorimotor gating. The effect of the synthetic cannabinoids on spontaneous locomotion was also measured by a video tracking analysis to assess potential cannabinoid-induced motor impairment. Results showed that, similarly to JWH-018, systemic administration of JWH-018-R compounds inhibits sensorimotor and PPI responses at lower doses (0.01-0.1 mg/kg) and reduced spontaneous locomotion at intermediate/high doses (1–6 mg/kg). All effects were prevented by the administration of the selective cannabinoid CB1 receptor antagonist/inverse agonist AM-251 thus confirming a CB1 receptor-mediated action. Finding that lower doses of JWH-018-R compounds selectively impair sensorimotor and PPI responses without affecting locomotion should be carefully considered to better understand the potential danger that halogenated-derivatives of JWH-018 may pose to public health, with particular reference to decreased performance in driving and hazardous works.

MDPV is a synthetic cathinone illegally marketed and consumed for its psychostimulant effects, which are similar to those produced by cocaine, amphetamines, and MDMA. Clinical reports indicate that MDPV produces euphoria, increases alertness, and at high doses causes agitation, psychosis, tachycardia and hypertension, hallucinations, delirium, hyperthermia, rhabdomyolysis, and even death. In rodents, MDPV reproduces the typical physiological effects of psychostimulant drugs, demonstrating greater potency than cocaine. Nevertheless, its role in aggressive behavior has been reported but not yet experimentally confirmed. Therefore, the aim of this study was to evaluate the effects of acute and repeated MDPV (0.01–10 mg/kg i.p.) administration on aggressive behavior in mice and to compare them with those of cocaine (0.01–10 mg/kg i.p.) administration. To this purpose, the resident–intruder test in isolated mice and the spontaneous and stimulated aggressiveness tests for group-housed mice were employed. The present study shows for the first time that MDPV enhances aggressive behavior and locomotion in mice with greater potency and efficacy than cocaine treatment. Moreover, the aggressive and locomotor responses are enhanced after repeated administration, indicating that a sensitization mechanism comes into play. These results, although from preclinical investigation, are suggestive that human MDPV intake could be a problem for public health and the criminal justice system. Thus, investigation by police officers and medical staff is needed to prevent interpersonal violence induced by the consumption of synthetic cathinones.

Clinical reliability of self-reported data for alcohol, caffeine, and nicotine consumptions is lacking, particularly in adolescents.To compare a self-report questionnaire and hair analysis to assess the reliability and effectiveness of the self-report.A cross-sectional study on 14-15-year-old Italian students (n = 874, 38% males, 62% females) was performed comparing self-reported data to hair analysis. The latter quantified hair concentrations of caffeine, nicotine, cotinine, ethyl glucuronide (EtG), and fatty acid ethyl esters (FAEEs) using mass spectrometry.Concordance between self-report and hair testing ranged from good to poor across substances and levels of use: poor for heavy alcohol intake (EtG: k = 0.36, 20 positive cases by hair analysis, false negative by self-report, 2.3% of total sample; FAEE k = 0.31, 25 positive cases, 2.9% of total sample); fair to poor for active smokers (k = 0.40, 125 positive cases, 14.3% of total sample); and moderate for caffeine (k = 0.57, 56 positive cases, 6.4% of total sample).Epidemiological studies on alcohol, caffeine, and nicotine consumption in adolescents may benefit from the inclusion of toxicological analysis on hair samples to overcome the under-reporting phenomenon of questionnaires and detect more cases of problematic substance use.

JWH-250 and JWH-073 are two synthetic cannabinoid agonists with nanomolar affinity at CB1 and CB2 receptors. They are illegally marketed within "herbal blend" for theirs psychoactive effects greater than those produced by Cannabis. Recently, we analyzed an "herbal" preparation containing a mixture of both JWH-250 and JWH-073. The present study was aimed at investigating the in vitro and in vivo pharmacological activity of JWH-250 and JWH-073 in male CD-1 mice. In vitro competition binding experiments performed on mouse and human CB1 and CB2 receptors revealed a nanomolar affinity and potency of the JWH-250 and JWH-073. In vivo studies showed that JWH-250 and JWH-073, administered separately, induced a marked hypothermia, increased pain threshold to both noxious mechanical and thermal stimuli, caused catalepsy, reduced motor activity, impaired sensorimotor responses (visual, acoustic and tactile), caused seizures, myoclonia, hyperreflexia and promote aggressiveness in mice. Moreover, microdialysis study in freely moving mice showed that systemic administration of JWH-250 and JWH-073 stimulated dopamine release in the nucleus accumbens in a dose-dependent manner. Behavioral, neurological and neurochemical effects were fully prevented by the selective CB1 receptor antagonist/inverse agonist AM 251. Co-administration of ineffective doses of JWH-250 and JWH-073 impaired visual sensorimotor responses, improved mechanical pain threshold and stimulated mesolimbic DA transmission in mice, living unchanged all other behavioral and physiological parameters. For the first time the present study demonstrates the overall pharmacological effects induced by the administration of JWH-250 and JWH-073 in mice and it reveals their potentially synergistic action suggesting that co-administration of different synthetic cannabinoids may potentiate the detrimental effects of individual compounds increasing their dangerousness and abuse potential.

This is a protocol for a Cochrane Review (Intervention). The objectives are as follows: To assess the clinical effectiveness and safety of the combined Antiretroviral Treatment containing ABC relative to ABC‐sparing cART. A subgroup comparison will be performed in studies comparing Antiretroviral Treatment containing ABC relative to TDF containing cART.

A sensitive and selective HPLC–MS/MS method has been developed for the first time for the anal- ysis of Δ9-tetrahydrocannabinol (the most important active cannabinoid) and its hydroxylated and carboxylated metabolites in human Dried Blood Spots (DBSs). The simultaneous determination of A sensitive and selective HPLC–MS/MS method has been developed for the first time for the anal- ysis of Δ9-tetrahydrocannabinol (the most important active cannabinoid) and its hydroxylated and carboxylated metabolites in human Dried Blood Spots (DBSs). The simultaneous determination of Δ9- tetrahydrocannabinol and its two main metabolites allows assessing the time elapsed after the drug intake and distinguishing between acute or former consumption. This is an important information in specific contexts such as “on street” controls by police forces. DBSs have been chosen as the optimal biological matrix for this kind of testing, since they provide information on the actual state of intoxica- tion, without storage and transportation problems usually associated with classical blood testing. The analysis is carried out on a C8 reversed phase column with a mobile phase composed of 0.1% formic acid in a water/methanol mixture and an electrospray ionisation (ESI) source, coupled to a triple quadrupole mass spectrometer. The method was validated according to international guidelines, with satisfactory results in terms of extraction yields, precision, stability and accuracy. Application to real DBS samples from Cannabis abusers gave reliable results, thus confirming the methodology suitability for roadside testing 9- tetrahydrocannabinol and its two main metabolites allows assessing the time elapsed after the drug intake and distinguishing between acute or former consumption. This is an important information in specific contexts such as “on street” controls by police forces. DBSs have been chosen as the optimal biological matrix for this kind of testing, since they provide information on the actual state of intoxica- tion, without storage and transportation problems usually associated with classical blood testing. The analysis is carried out on a C8 reversed phase column with a mobile phase composed of 0.1% formic acid in a water/methanol mixture and an electrospray ionisation (ESI) source, coupled to a triple quadrupole mass spectrometer. The method was validated according to international guidelines, with satisfactory results in terms of extraction yields, precision, stability and accuracy. Application to real DBS samples from Cannabis abusers gave reliable results, thus confirming the methodology suitability for roadside testing.

Objectives: The efficacy of abacavir/lamivudine has been reported to be inferior to tenofovir/emtricitabine. Several randomized clinical trials (RCTs) investigated the effectiveness and safety of abacavir/lamivudine and tenofovir/emtricitabine combined antiretroviral treatment (cART) and we have reviewed the available evidence. Design: Systematic review and meta-analysis of RCTs using standard Cochrane Collaboration methodologies. Methods: We calculated risk ratios (RRs) with 95% CIs. The primary outcome was the rate of patients with viral load (VL) below the pre-defined cut-off at 48 weeks and/or at 96 weeks. Where available, results were analysed according to VL screening levels (,100000 or .100000 copies/mL) with conventional meta-analytical pooling by subgroups and meta-regression. Results:Meta-analytical pooling of RCTs with a direct comparisonof abacavir/lamivudine and tenofovir/emtricitabine according to baseline VL at 48 weeks (six trials, 4118 patients) showed that the proportions of subjects with VL ,50 copies/mL were similar in the overall comparison (RR 0.98; 95% CI 0.94‐1.03), in the low baseline VL strata (RR 1.01; 95% CI 0.99‐1.03) and in the high baseline VL strata (RR 0.96; 95% CI 0.90‐1.03). Meta-regression analysis at 48 weeks confirms the results of subgroup analysis. Similar virological results were found at 96 weeks (four trials, 2003 patients). Differences in the occurrence of adverse events requiring discontinuation of treatment favoured tenofovir recipients (RR 1.26; 95% CI 0.99‐1.61), but this difference, mostly related to suspected abacavir hypersensitivity reaction, was not statistically significant. Conclusions: Ourcumulative, cross-sectional data suggest a similar virological efficacy of abacavir/lamivudine and tenofovir/emtricitabine regardless of the baseline VL.

RATIONALE New psychoactive substances (NPSs) are rapidly spreading worldwide, and forensic laboratories are often requested to identify new substances for which no reference standards or analytical data are available. This article describes an analytical approach that was adopted in Italy by a few collaborative centres of the Italian Early Warning System for Drugs, which has contributed many alerts for the identification of different classes of NPSs in the last 24 months. METHODS Seized crystals and powders were initially analysed via single quadrupole gas chromatography/mass spectrometry (GC/MS), followed by liquid chromatography/high-resolution mass spectrometry (LC/HRMS) in the positive electrospray ionisation (ESI) mode at 100 000 full width at half maximum resolution (FWHM) without fragmentation to elucidate the elemental compositions of unknown molecules. Different fragmentation voltages during LC/HRMS were applied to study the accurate masses of the obtained characteristic fragments. Nuclear magnetic resonance (NMR) analyses were performed to identify specific isomers when necessary. RESULTS Some interesting examples of unknown NPSs from seizures later identified in our laboratories are reported, with special focus on those cases where analytical standards were not available during analyses. These cases include cathinones, such as 3-methylmethcathinone (3-MMC), methylone, bk-MBDB (butylone), 4-methylethcathinone (4-MEC), flephedrone, methylenedioxypyrovalerone (MDPV) and pentedrone, methoxetamine, apinaca or AKB48, benzydamine, meta-chlorophenylpiperazine (m-CPP), 5-MeO-N,N-dialkyl tryptamines, such as 5-MeO-DALT and 5-MeOMIPT, benzofurans, such as 6-APB and 4-APB, and diphenidine (identified for the first time in Europe). CONCLUSIONS The identification of NPSs in confiscated materials was successfully achieved via GC/MS coupled with LC/HRMS and, in a few cases, NMR analyses. The availability of GC/MS libraries is of great assistance in the identification of new drugs. Alternatively, the study of characteristic molecule fragments combined with the determination of their accurate masses can be a useful approach to identify unknown samples not previously analysed. Copyright © 2014 John Wiley & Sons, Ltd.

Cannabis derivatives are among the most widely used illicit substances among young people. The addictive potential of delta-9-tetrahydrocannabinol (THC), the major active ingredient of cannabis is well documented in scientific literature. However, the consequence of THC exposure during adolescence on occurrence of addiction for other drugs of abuse later in life is still controversial. To explore this aspect of THC pharmacology, in the present study, we treated adolescent rats from postnatal day (PND) 35 to PND-46 with increasing daily doses of THC (2.5-10mg/kg). One week after intoxication, the rats were tested for anxiety-like behavior in the elevated plus maze (EPM) test. One month later (starting from PND 75), rats were trained to operantly self-administer heroin intravenously. Finally, following extinction phase, reinstatement of lever pressing elicited by the pharmacological stressor, yohimbine (1.25mg/kg) was evaluated. Data revealed that in comparison to controls, animals treated with chronic THC during adolescence showed a higher level of anxiety-like behavior. When tested for heroin (20μg per infusion) self-administration, no significant differences were observed in both the acquisition of operant responding and heroin intake at baseline. Noteworthy, following the extinction phase, administration of yohimbine elicited a significantly higher level of heroin seeking in rats previously exposed to THC. Altogether these findings demonstrate that chronic exposure to THC during adolescence is responsible for heightened anxiety and increased vulnerability to drug relapse in adulthood.

Background: Monitoring consumption by population surveys (PS) is an important way to challenge the spread of illicit drugs (ID). To improve the information, we explored a complementary method, particularly wastewater analysis (WWA). Methods: We estimated the prevalence of use by PS, and the consumption by WWA, of cocaine, opioids, cannabis, methamphetamine and MDMA (ecstasy) from 2010 to 2014 in Italy and compared the results. Results: According to PS, cannabis and cocaine were the ID most used in Italy (last month prevalence 3.0% and 0.43% respectively in 2010) followed by opioids (0.17%) and amphetamines (0.14%). WWA gave similar findings, with cannabis consumption (4.35 g THC/day/1000 inhabitants) exceeding cocaine (0.78 g), heroin (0.092 g), methamphetamine and MDMA (0.103 g). The time trend investigated by PS showed significant decreases for all ID from 2010 to 2012. WWA also indicated a reduction of consumption for methamphetamine (p

Drug abuse assessment methods based on measuring illicit substances in waste waters are consolidated. The approach of ambient air monitoring looks questionable, nonetheless it can be explored if the variables determining the drug burdens are accounted for, or suitable co-contaminants are adopted to normalize concentrations to environmental and human contours. The general approach linking the airborne drug concentrations to consumption is presented and the case of cocaine is discussed according to measurements conducted in Italy. The cocaine/nicotine concentration ratio, identified as the most suitable tool, fitted well with anti-drug Police operations and people noticed for drug-related crimes, and with the abuse prevalence estimated in the cities investigated. According to that, the conversion factors of drug concentrations into prevalence estimates seem assessable, provided sufficient databases over space and time are collected. Further investigations are necessary to understand if airborne drugs cause adverse sanitary effects.© 2013 Elsevier Ltd. All rights reserved.

New psychoactive substances (NPS) are a heterogeneous and rapidly evolving class of molecules available on the global illicit drug market (e.g smart shops, internet, “dark net”) as a substitute for controlled substances. The use of NPS, mainly consumed along with other drugs of abuse and/or alcohol, has resulted in a significantly growing number of mortality and emergency admissions for overdoses, as reported by several poison centers from all over the world. The fact that the number of NPS have more than doubled over the last 10 years, is a critical challenge to governments, the scientific community, and civil society (UNODC, World Drug Report, 2014; EMCDDA, European Drug Report 2014: Trends and developments). The chemical structure (phenethylamines, piperazine, cathinones, tryptamines, synthetic cannabinoids) of NPS and their pharmacological and clinical effects (hallucinogenic, anesthetic, dissociative, depressant) help classify them into different categories. In the recent past, 50% of newly identified NPS have been classified as synthetic cannabinoids followed by new phenethylamines (17%)(WDR, 2014). Besides peripheral toxicological effects, many NPS seem to have addictive properties. Behavioral, neurochemical, and electrophysiological evidence can help in detecting them. This manuscript will review existing literature about the addictive and rewarding properties of the most popular NPS classes: cannabimimetics (JWH, HU, CP series) and amphetamine-like stimulants (amphetamine, methamphetamine, methcathinone and MDMA analogues). Moreover, the review will include recent data from our lab which links JWH-018, a CB1 and CB2 agonist more potent than Δ9-THC, to other cannabinoids with known abuse potential, and to other classes of abused drugs that increase dopamine signaling in the Nucleus Accumbens (NAc) shell. Thus the neurochemical mechanisms that produce the rewarding properties of JWH-018, which most likely contributes to the greater incidence of dependence associated with “Spice” use, will be described (De Luca et al., 2015a). Considering the growing evidence of a widespread use of NPS, this review will be useful to understand the new trends in the field of drug reward and drug addiction by revealing the rewarding properties of NPS, and will be helpful to gather reliable data regarding the abuse potential of these compounds.

It is well known that an impairment of learning and memory function is one of the major physiological effects caused by natural or synthetic cannabinoid consumption in rodents, nonhuman primates and in humans. JWH-018 and its halogenated derivatives (JWH-018-Cl and JWH-018-Br) are synthetic CB 1 /CB 2 cannabinoid agonists, illegally marketed as “Spice” and “herbal blend” for their Cannabis-like psychoactive effects. In the present study the effects of acute exposure to JWH-018, JWH-018-Cl, JWH-018-Br (JWH-018-R compounds) and Δ 9 -THC (for comparison) on Novel Object Recognition test (NOR) has been investigated in mice. Moreover, to better characterize the effects of JWH-018-R compounds on memory function, in vitro electrophysiological and neurochemical studies in hippocampal preparations have been performed. JWH-018, JWH-018-Cl and JWH-018-Br dose-dependently impaired both short- and long-memory retention in mice (respectively 2 and 24 h after training session). Their effects resulted more potent respect to that evoked by Δ 9 -THC. Moreover, in vitro studies showed as JWH-018-R compounds negatively affected electrically evoked synaptic transmission, LTP and aminoacid (glutamate and GABA) release in hippocampal slices. Behavioral, electrophysiological and neurochemical effects were fully prevented by CB 1 receptor antagonist AM251 pretreatment, suggesting a CB 1 receptor involvement. These data support the hypothesis that synthetic JWH-018-R compounds, as Δ 9 -THC, impair cognitive function in mice by interfering with hippocampal synaptic transmission and memory mechanisms. This data outline the danger that the use and/or abuse of these synthetic cannabinoids may represent for the cognitive process in human consumer.

A key issue in the prevention and control of Sexually Transmitted Diseases (STD) is to provide access to health centres, and in diagnosing and treating STD. The present study is aimed to assess the prevalence of sexually transmitted diseases (STDs) and Hepatitis C virus (HCV) in a population of immigrant female sex workers (FSWs). We conducted a cross sectional survey of FSWs working in Verona, North-eastern Italy. Screening test included serology for STDs [including Human Immunodeficiency Virus (HIV), syphilis and Hepatitis B virus (HBV)] and hepatitis C virus (HCV). Sixteen out of 345 (4.6%) street FSWs screened during 1999-2007 resulted positive for HIV, 12 (3.5%) were positive for HBsAg, 7 (2.0%) were positive for syphilis serological test, and 3 (0.9%) were positive for HCV. Comparison of the prevalence data between women from Africa (286/345, 82.8%) and other countries showed no statistical difference for HIV infection (R.R. 1.44; 95% CI, 0.34-6.19) and for presence of HBsAg (R.R. 2.27; 95% CI, 0.30-17.24). The positivity of syphilis serologic tests had a lower prevalence among African FSWs (mostly coming from Nigeria) than among FSWs from Eastern Europe (57/345, 16.5%). This difference was statistically significant (R.R. 0.03; 95% CI, 0.00-0.28). The prevalence of HIV infection increased with age (p=0.04, by chi2 for trend analysis), but not with the time worked as sex workers in Italy. Moreover, the presence of any of the screened infections was predictable by both age and earlier time of immigration by way of logistic multivariable regression. The prevalence of HIV and HBsAg was higher in the whole analyzed cohort compared to the general population; prevalence of syphilis was significantly higher in FSWs from Eastern Europe than in FSWs from Africa. HCV prevalence remains low among non intravenous drug abuser FSWs. The data offers a starting point to address targeted intervention that would prevent FSWs acquiring and transmitting STDs.

A B S T R A C T A novel method based on microemulsion electrokinetic chromatography (MEEKC) with diode array detection (DAD) for rapid determination of caffeine in commercial and clandestine stimulants, known as ‘‘energy drinks’’ and ‘‘smart drugs’’, is described. Separations were carried out in 50 cm � 50 mm (ID) uncoated fused silica capillaries. The optimized buffer electrolyte was composed of 8.85 mM sodium tetraborate pH 9.5, SDS 3.3% (w/v), n-hexane 1.5% (v/v) and 1-butanol 6.6% (v/v). Separations were performed at a voltage of 20 kV. Sample injection conditions were 0.5 psi, 3 s. Diprofilline was used as internal standard. The determination of the analytes was based on the UV signal recorded at 275 nm, corresponding to the maximum wavelength of absorbance of caffeine, whereas peak identification and purity check was performed on the basis of the acquisition of UV radiation between 200 and 400 nm wavelengths. Under the described conditions, the separation of the compounds was achieved in 6 min without any interference from the matrix. Linearity was assessed within a caffeine concentration range from 5 to 100 mg/mL. The intra-day and inter-day precision values were below 0.37% for migration times and below 9.86% for peak areas. The present MEEKC method was successfully applied to the direct determination of caffeine in smart drugs and energy drinks.

The epidemiology of Candida infection in intensive care units (ICUs) and the management strategies for such infections in non-neutropenic intensive care patients are discussed in this review. Candida species are one of the leading causes of nosocomial bloodstream infections and a significant cause of morbidity in patients admitted to the ICU. Prophylactic, pre-emptive and empiric treatment strategies for Candida infections have been explored in ICU patients. Routine prophylaxis should not be administered to the whole population of ICU patients, because the concerns about the selection of azole-resistant Candida strains or the induction of resistance are justified. Treatment of fungal infections is now possible with newer antifungal agents, including newer azoles (e.g., voriconazole, posaconazole) and echinocandins (e.g., micafungin, anidulafungin). However, there is a critical need for improvement in diagnosis of invasive Candida infection in order to provide clinicians the opportunity to intervene earlier in the diseases course.

Although home Internet access has continued to increase, little is known about actual usage patterns in homes. This nationally representative study of over 4,700 Italian households with children measured computer and Internet use of each family member across 3 months. Data on actual computer and Internet usage were collected by Nielsen//NetRatings service and provide national baseline information on several variables for several age groups separately, including children, adolescents, and adult men and women. National averages are shown for the average amount of time spent using computers and on the Web, the percentage of each age group online, and the types of Web sites viewed. Overall, about one-third of children ages 2 to 11, three-fourths of adolescents and adult women, and over four-fifths of adult men access the Internet each month. Children spend an average of 22 hours/month on the computer, with a jump to 87 hours/month for adolescents. Adult women spend less time (about 60 hours/month), and adult men spend more (over 100). The types of Web sites visited are reported, including the top five for each age group. In general, search engines and Web portals are the top sites visited, regardless of age group. These data provide a baseline for comparisons across time and cultures.

The efficacy of granulocyte macrophage colony-stimulating factor (GM-CSF) to enhance the immune response to hepatitis B virus vaccine has been object of several reports. We searched for randomized controlled clinical trials comparing GM-CSF given concomitantly to hepatitis B virus vaccine to vaccine given alone or with placebo. Data on rates of seroconversion (anti-HBs titers >10 IU/ml) from 13 studies (734 subjects) produced combined estimates that favored GM-CSF as compared to controls: rate ratio after a single immunization was 1.54 [95% confidence interval (CI), 1.04-2.27] and 1.20 (95% CI, 1.02-1.42) at the end of the vaccination cycle. Using a logistic approach a significant dose/response effect of GM-CSF was seen. Moreover, in renal failure patients who have responded to the vaccine, GM-CSF increased anti-HBs titers. Our findings suggest that GM-CSF induced a significant effect in terms of response rate and achievement of an earlier seroconversion to the vaccine in the overall populations examined, in renal failure patients and in healthy individuals.

γ-Hydroxybutyric acid (GHB) spread for recreational purposes or as "rape drug" represents a hard issue for forensic toxicologists due to its endogenous nature. It is clear that an actual and reliable discrimination between basal and exogenous levels is mandatory to achieve a correct evaluation of conscious/unconscious administration. This research aimed to study the GHB baseline in hair samples, collected from 150 volunteers, non-consumers of any drugs of abuse, in order to evaluate if a generic cut-off value could be accepted, also focusing on potential influences of gender and age. The analysis consisted of an overnight incubation with NaOH at 56 °C, liquid-liquid extraction with ethylacetate and trimethylsylil derivatization. Detection was carried out through gas chromatography-mass spectrometry in single ion monitoring (m/z 233, 234, 147 for GHB; m/z 239, 240 and 147 for GHB-d6). The endogenous amount in "blank" hair was estimated by the standard addition method. Concentration range was 0.279-2.839 ng/mg. In males, the average GHB levels were higher than in females (0.829 vs 0.596 ng/mg, respectively), especially in the first age category (30 years, 1.008 vs 0.606 ng/mg, respectively). Age influences on GHB levels seemed to be different among the two sexes: in male population concentrations were higher30 (1.008 ng/mg) and similar in the other age ranges (0.762 ng/mg, 30-50; 0.763 ng/mg,50); in female, quite similar levels were registered throughout all the age categories (0.606 ng/mg,30; 0.536 ng/mg, 30-50; 0.691 ng/mg,50). Further study should be performed on GHB physiology in order to better understand these differences among ages and genders. Moreover, we demonstrated that for hair analysis a cut-off reference value is not strictly mandatory, underlining the great interpretative valence of segmental analysis.

The “3R concept”, first introduced by Russell and Burch's 1959 book “The principles of humane experimental technique”, is a widely accepted ethical framework for conducting scientific experiments on animals: Replacement (use of non-animal methods), Reduction (methods which reduce the number of animals used) and Refinement (methods which improve animal welfare). In particular, the refinement priciple refers to the improvement of breeding and of experimental procedures, aiming to minimise actual or potential pain and to improve animal welfare in those situations where animal usage is unavoidable. In animal experimental studies high volumes of blood or plasma are needed for an accurate analysis of the compounds of interest. Because of this, often the animal sacrifice is necessary for small ones, such as rats and mice, which are also the most used in all laboratories. A possible alternative and innovative sampling in this research field is represented by the Dried Blood Spot (DBS) technique, that needs very small amount of blood. The extension of the DBS procedure to the animal model testing, sampling only a few blood microliters from the rat or mouse tail, would leave the animal alive, allowing to perform serial samplings on the same subject. Other main advantages are represented by a minimally invasive withdrawal, easier storage and transportation issues, a better stability of the analysed compounds and more feasibility of post-sampling procedures. The purpose of this work is the development and validation of a rapid and reliable analytical strategy for the determination of cannabinoids in DBS, obtained from mice treated with Δ9-tetrahydrocannabinol within an immunological research study.

An advanced analytical method based on liquid chromatography–tandem mass spectrometry (LC–MS/MS), has been developed for the identification and determination in hair of Δ9- tetrahydrocannabinol together with its major metabolite 11-nor-9-carboxy-Δ9-tetrahydrocannabinol. Since the latter is formed endogenously, it allows the assessment of chronic use excluding passive exposure to Cannabis. The sample pre-treatment procedure is based on a feasible incubative extrac- tion followed by a liquid–liquid extraction step. Chromatographic separation was performed using a reversed-phase column and gradient elution with a formic acid/acetonitrile/water mobile phase. The lim- its of quantitation and of detection were 3 pg/mg and 1 pg/mg, respectively, for both analytes. The method was successfully applied to the analysis of hair samples from Cannabis abusers; the analyte concentrations found ranged from 55 to 100 pg/mg for Δ9 -tetrahydrocannabinol and from 5 to 10 pg/mg for 11-nor-9- carboxy-Δ9 -tetrahydrocannabinol. Accuracy studies also gave satisfactory results (recovery > 87%), thus confirming the suitability of the assay for chronic consumption monitoring.

Ketamine (2-(2-chlorophenyl)-2-(methylamino)cyclohexanone) is a human and veterinary anaesthetic and analgesic with hallucinogenic properties. For this reason it is illegally used for recreational purposes and included in the controlled substance schedules of most countries. The compound possesses a stereogenic asymmetric carbon atom and can exist as a pair of enantiomers. Pharmacological studies have demonstrated that the S-(+)-enantiomer of ketamine is about three times more potent than the R-(–)-enantiomer as an analgesic. Currently, most ketamine formulations are racemic, however enantiopure S-ketamine is also available on the market under the brand name "Ketanest S". On the other hand, even if ketamine hallucinogenic potency is still largely unclear, some authors claim that the R-enantiomer is the most potent. Therefore it is interesting and useful to investigate the exact enantiomeric composition of ketamine, both when used as a conventional medicine and when abused as an illicit drug. Ketamine eudysmic ratio could be also studied as a marker of origin to discriminate between authorised industrial manufacturing and illicit “home made” synthesis. Moreover, ketamine chiral analysis in biological fluids can provide information on the intended use of the substance, considering that its racemisation in the body seems to be very low. The aim of this study is to test different polysaccharide-based stationary phases [1] and different chromatographic conditions to obtain a good ketamine enantioseparation by liquid chromatography with native fluorimetric detection (HPLC-FL). The method is under validation and promising to be applied for the chiral analysis of ketamine in different matrices, such as prescribed drugs, police confiscations and biological fluids, with the purpose of quality control analyses, detecting illicit use and contributing to the identification of the source of seized ketamine batches.

Herbal incense mixtures, of which “n-Joy” is perhaps one of the best known examples, have been available globally for the past two or three years. Although declared as incense and not for human consumption, herbal-type products are smoked as an apparently legal alternative to Cannabis to deliver the so-called “herbal high”. Several synthetic cannabinoid receptor agonist (cannabinomimetic) compounds have been identified in herbal-type products to date. The detected substances include the JWH series, developed as test leads in the investigation of drug-receptor interactions. A few of these compounds have been designated as controlled drugs in some countries. It is apparent, however, that as one set of compounds is claimed illegal, more variants that fall outside the current legislation take their place. Thus, there is an urgent need for comprehensive methods suitable for the analysis of synthetic cannabinoids, particularly in the fields of clinical and forensic toxicology. The present study deals with the analysis of illicit materials seized by police forces, in order to verify the presence of synthetic cannabinoids belonging to the JWH series, by structure identification strategies. A suspect sample of a herbal mix, named “WWW Aromatic Potpourri”, was preliminary analysed by Scientific Police of Bologna and a synthetic cannabinoid of the JWH family was identified by means of GC-MS techniques. Moreover, a MS/MS method optimised on a triple quadrupole mass spectrometer was implemented for the determination of the identified compound and possibly of other illicit substances. Mass spectrum data have been acquired in order to investigate the structural hypothesis and to provide m/z transitions, required for the development of an advanced LC-MS/MS analytical system, able to carry out also a quantitative study.

Ketamine ((R,S-2-(2-chlorophenyl)-2-methylamino)cyclohexanone) is a phencyclidine derivative used in human and veterinary clinical practice since 1970. It is a dissociative anaesthetic agent that induces also analgesia by non-opioid mechanisms. Ketamine causes a state of "dissociative anaesthesia", for this reason is used as a recreational drug and has been included in the controlled substance schedules of most countries. As an anaesthetic drug, Ketamine is commercially available as a racemic mixture however, (R)-Ketamine and (S)-Ketamine have significantly different pharmacodynamic activities: the therapeutic potency of (S)-Ket is 2-4 times greater than the that of the (R)-enantiomer, as regards anaesthetic and analgesic effects. Moreover, the post-hypnotic stimulatory properties and agitated behaviour are associated with (R)-Ket. On the other hand, even if Ketamine hallucinogenic potency is still largely unclear, some authors claim that the (R)-enantiomer is the most potent one. It has also been reported that (R)- and (S)-Ket have significantly different pharmacokinetic profiles. Thus, it is evident the need to provide analytical methods able to discriminate and simultaneously quantify both Ketamine enantiomers in biological matrices for pharmacokinetic, toxicological and forensic purposes. The aim of this study is the development of an analytical method for Ketamine chiral analysis in alternative and innovative biological fluids and tissues, such as Dried Blood Spots (DBSs), saliva and hair. Different chromatographic setups were tested to obtain a good enantioseparation by liquid chromatography with fluorimetric detection (HPLC-F). The assays are currently under validation and seem to be promising for a successfull Ketamine enantiomeric resolution and determination, in order to be applied to real biological samples.

Propofol is a short-acting hypnotic agent that is commonly used to induce and maintain anesthesia. Propofol abuse and its involvement in suicide deaths have increased in recent years, especially among healthcare personnel. An example is the suicide of a 61-year-old nurse found with a propofol drip in his left arm. We describe the postmortem concentration of propofol in various tissues (femoral and cardiac blood, bile, urine, brain, and liver) and in the drip. The toxicological analyses were performed through two analytical methods, differing in derivatization reaction and in instrumentation: silylation for gas chromatograph–mass spectrometer (GC–MS), as routinely performed in our laboratory for this kind of analyses (lower limits of quantification–LLOQ–in urine and blood: 0.3 and 5 ng/ml); for liquid chromatograph–tandem mass spectrometer (LC–MS/MS) an innovative azo-coupling derivatization (LLOQ: 0.0004 and 0.1 ng/ml). This latter produces an azo-derivative (molecular composition: C 18 H 22 ON 2 ; molecular weight: 282 Da) highly ionizable in electro-spray ion source, both in negative and positive ionizations. These two methods were compared to evaluate the effectiveness of this new LC–MS/MS analysis. An acidic hydrolysis (HCl 6 N, 100 °C, and 1 h) was performed for the biological samples (1 ml or 1 g) irrespective of the analytical method applied. The drip content was extracted adding phosphate buffer (pH 8) and a dichloromethane/ethylacetate 8:2 (v:v) mixture. Derivatization steps were: silylation with N , O -bis(trimethylsilyl)trifluoroacetamide (BSTFA) + tetramethylammonium hydroxide (TMAH) for GC–MS; regarding LC–MS/MS, azo-coupling reaction with the aryl-diazonium salt (0–5 °C, and 30 min). The analyses were achieved in selected-ion monitoring for GC–MS ( m/z , 235,250,73 propofol”; m/z , 252,267,27 propofol-d17) and in multiple reaction monitoring ([M−H] − : m/z 283→241,77, azo-propofol; m/z 299→251,77, azo-propofol-d17) for LC–MS/MS. Autopsy showed no significant findings. Propofol concentrations were (LC–MS/MS vs GC–MS, respectively): 15.1 vs 14.5 mg/ml, drip content; 7.11 vs 6.07 μg/ml, cardiac blood; 9.50 vs 7.19 μg/ml, femoral blood; 0.64 vs 1.07 μg/ml, bile; 0.042 vs 0.051 μg/ml urine; 4.93 vs 5.89 μg/g, brain; and 7.88 vs 6.80 μg/g, liver. These values are comparable with the ones described in literature for death by acute propofol intoxication; the drip content is compatible with a diluted formulation of propofol available in Italy (20 mg/ml injectable emulsion). The comparison shows an excellent fitting of the data (R 2 : 0.9362). Toxicological results proved the cause of death as acute propofol intoxication. Furthermore, the new LC–MS/MS method showed an excellent effectiveness and reliability when compared to the routinely used GC–MS method.

To better understand why some drug abuse treatment programs are more effective than others, USA research about organizational functioning and its role in the provision of treatment services was extended through a study of a delivery system in another country. The Texas Christian University (TCU) organizational functioning and readiness for change instrument (ORC) was translated into Italian and administered to 405 treatment program directors and staff from both public and private sectors in the Veneto Region of Northern Italy. Results indicated that the psychometric properties of the ORC in the USA and Italy are consistent. Some general differences in staff attributes were found between USA and Italian programs, but organizational climates were remarkably similar.

Cannabis is the most widely used illicit drug around the world and among the drugs of abuse (DoA) most frequently involved in street accidents. Therefore, it is important to strictly monitor Cannabis abuse. Until now, the most reliable biological matrix for this purpose is represented by blood (whole blood/plasma), which can provide useful information about drug consumption. However, its sampling and storage require complicated and time consuming precautions, such as centrifugation and refrigeration or freezing, in order to preserve analyte stability. To overcome these disadvantages the use of Dried Blood Spots (DBSs) represents a valid alternative to the “classical” blood sampling. This innovative biological matrix reproduces the composition of whole blood but its pre-treatment procedure is faster and more feasible. DBSs can be stored for months at room temperature without any appreciable sample degradation, since most enzymatic and non-enzymatic reactions are stopped by the loss of water. The aim of this work is the specific evaluation of cannabinoid stability in DBSs, namely of Δ9 tetrahydrocannabinol (THC), the most important psychoactive cannabinoid, and of its two main metabolites 11-hydroxy-Δ9-tetrahydrocannabinol (THC-OH) and 11-nor-9-carboxy-Δ9-tetrahydrocannabinol (THC-COOH). For this study, blank spiked DBSs were stored at room temperature for up to 3 months after sampling and then analysed by ESI-LC-MS/MS. The sample pre-treatment, based on solvent extraction, provides good extraction yields and selectivity. Preliminary results are very satisfactory, since only minimal differences were observed between the samples analysed immediately after spotting and those analysed after storage. Further assays are in progress to confirm these results and to assess cannabinoid stability in DBSs over longer periods of time.

Driving under the influence of Cannabis (DUI) has become a growing concern. Studies investigating the impact of DUI on traffic safety have shown evidence that, during the acute period of Cannabis intoxication, Cannabis diminishes driving faculties greatly increasing the risk of car accidents. However, the actual state of Cannabis intoxication is not easily assessed, particularly in specific contexts such as roadside testing. Until now, the most reliable biological matrix for this purpose in represented by blood but its sampling is invasive and requires a sanitary environment and subsequent treatments or storage precautions, such as centrifugation, refrigeration or freezing. The time lapse between the individuation of a possible intoxication and the blood sampling is very important, since a long delay can mean that, in the meantime, the drug blood levels physiologically decrease under the cut-off value. To overcome this disadvantage, Dried Blood Spots (DBSs) can be a valid alternative to the normal blood sampling by venipuncture. In fact this innovative biological matrix can be obtained after a fast and easy sampling, does not need any particular storage nor transportation precaution and is stable over time. To strictly monitor Cannabis consumption, an original LC-MS/MS method has been developed for the analysis of Cannabinoids in DBSs. Attention has been paid to the determination of Δ9- tetrahydrocannabinol (or THC, Figure 1a), the main psychoactive compound whose presence in blood can be taken as a marker of recent exposure, and its two main metabolites 11-hydroxy-Δ9-tetrahydrocannabinol (THC-OH, Figure 1b) and 11-nor-9-carboxy-Δ9-tetrahydrocannabinol. In particular the intermediate hydroxylated metabolite (THC-OH) is pharmacologically active and, at the same time, it is a marker of recent Cannabis intake, since it appears in blood about 13 minutes after consumption and has a relatively short half-life. The carboxylated metabolite (THC-COOH) is not psychoactive and has a very long half-life up to one month for chronic users. For this reason the presence of this latter compound alone in blood cannot be taken as a marker of recent exposure. Is evident, therefore, that the simultaneous monitoring of all the three analytes is necessary to outline pharmacokinetic and toxicokinetic state of abusers and contribute to the assessment of psychophysical state, distinguishing between acute or former consumption. The analytical method developed has been fully validated and applied to real DBS samples from Cannabis abusers with satisfactory results, thus confirming the methodology suitability for roadside testings.

Drug abuse is a severe social problem, expecially among young people, which can cause devastating impacts on crime rate and healthcare expenses. Ketamine (2-(2-chlorophenyl)-2-(methylamino)cyclohexan-1-one) is an emerging drug of abuse often easily accessed in the black market and erroneously perceived by young subjects as a “safe” and “clean” drug. Ketamine was marketed as an anaesthetic drug and at sub-therapeutic doses possesses psychedelic properties including cataleptic state with stupor, sedation and possible amnesia, which can lead to drug-facilitated sexual assaults. This, together with the recent increase in ketamine abuse and ketamine-related death reports, is arousing the attention of the authorities as well as that of the scientific community. Detection and quantitation of residual illicit drugs by the analysis of blood, urine and oral fluid are often employed in forensic and clinical settings; on the other hand, hair monitoring can reveal a retrospective reflection of drug abuse, providing a wider detection window, therefore this strategy can be a useful tool in chronic drug abuse monitoring. The goal of this study is the simultaneous analysis of ketamine and its major active metabolite norketamine (2-amino-2-(2-chlorophenyl)cyclohexan-1-one) in hair samples by means of HPLC separation, using a C18 column as the stationary phase and a mobile phase consisting of an acetonitrile/acidic aqueous buffer mixture. Sample preparation includes an acidic extractive incubation, followed by L/L extraction with organic solvents. Quantitative determination is achieved using detection means such as spectrofluorimetry (F), that exploits the native fluorescence of the analytes, and tandem mass spectrometry (MS/MS). Since the obtained results are promising, after validation the method will be applied for the monitoring in hair of chronic ketamine abusers.

In a meta-analysis of 10 studies, the BACTEC 960/MGIT and BACTEC 460 systems showed a sensitivity and specificity in detecting mycobacteria (1,381 strains from 14,745 clinical specimens) of 81.5 and 99.6% and 85.8 and 99.9%, respectively. Combined with solid media, the sensitivity of the two systems increased to 87.7 and 89.7%, respectively.

Purpose: Past reports and meta-analyses indicate that fluoroquinolones are highly effective in preventing Gram-negative infections in neutropenic cancer patients, but offer inadequate coverage for Gram-positive infections. We evaluated by meta-analysis the efficacy of the addition of antimicrobial agents with enhanced Gram-positive activity to prophylaxis with quinolones. Materials and Methods: Randomized trials comparing fluoroquinolones alone (ciprofloxacin, ofloxacin, pefloxacin, or norfloxacin) with fluoroquinolone in combination with Gram-positive prophylaxis (rifampin, vancomycin, amoxicillin, roxithromycin, or penicillin) were retrieved. We pooled relative risks (RRs) using a fixed-effects model. Results: Nine trials (1,202 patients) published between 1993 and 2000 meet inclusion criteria. Compared with fluoroquinolone alone, Gram-positive prophylaxis reduced total bacteremic episodes (RR, 1.54; 95% CI, 1.26 to 1.88), streptococcal infections (RR, 2.20; 95% CI, 1.44 to 3.37), coagulase-negative staphylococcal infections (RR, 1.46; 95% CI, 1.04 to 2.04), and rate of febrile patients (RR 1.08; 95% CI, 1.00 to 1.16). Occurrence of clinically documented infections, unexplained fever, and infectious mortality was similar in the two groups. The addition of Gram-positive prophylaxis, however, significantly increased side effects (RR, 0.46; 95% CI, 0.28 to 0.76). Rifampin use resulted in a higher incidence of undesirable effects. Conclusion: Considering the lack of cut-clear benefit on some parameters of morbidity and mortality, routine use of Gram-positive prophylaxis is not advisable. This strategy, however, should be particularly valuable in subgroups of patients at high risk of streptococcal infection (eg, those with severe and prolonged neutropenia or mucositis, and those receiving cytarabine). Problems of tolerability and the potential for the emergence of resistant microorganisms should be considered when prescribing prophylaxis with enhanced Gram-positive activity to neutropenic patients.

Sputum induction is a simple and noninvasive procedure for Pneumocystis carinii pneumonia (PCP) diagnosis in human immunodeficiency virus-1-positive patients, although less sensitive than bronchoalveolar lavage (BAL). In order to obtain an overview of the diagnostic accuracy of sputum induction, a systematic review and meta-analysis of studies reporting the comparative sensitivity and specificity of BAL (the "gold standard") and sputum induction was performed. The odds ratio and related 95% confidence interval were calculated using summary receiving operating characteristic curves as well as fixed-effect and random-effect models. Based on pooled data, the negative and positive predictive values were calculated for a range of PCP prevalence using a Bayesian approach. Seven prospective studies assessed the comparative accuracy of BAL and sputum induction. On the whole, sputum induction demonstrated 55.5% sensitivity and 98.6% specificity. The sensitivity of sputum induction was significantly higher with immunofluorescence than with cytochemical staining (67.1 versus 43.1%). In settings of 25-60% prevalence of PCP, the positive and negative predictive values ranged 86-96.7 and 66.2-89.8, respectively, with immunofluorescence, and 79-94.4 and 53-83.5% with cytochemical staining. In conclusion, in a setting of low prevalence of Pneumocystis carinii pneumonia, sputum induction, particularly with immunostaining, appears to be adequate for clinical decision-making.

Background and Objectives Adolescence is a time during which not only gambling, but also tobacco, alcohol, and illicit drug use, usually begin. The purpose of this paper is to provide an updated review of the literature on gambling and its associations with tobacco, alcohol, and illicit drug use among US youth. Methods An electronic literature search of PubMed and PsycInfo was conducted for studies since 2000 using the keywords “smoking,” “tobacco,” “nicotine,” “cigarette,” “gambling,“ “adolescence,” “adolescent,“ “alcohol,” and “substance use.” Ten articles with unique adolescent samples were located. Because the articles varied in regard to definitions of gambling, tobacco, alcohol, and drug use, we provide a qualitative review of included studies. Results Gambling prevalence rates ranged from 44.3% to 68% in national telephone-based surveys, from 24.4% to 86% among students in school-based surveys, and from 22.5% to 47.4% in surveys of convenience samples. Significant associations were reported between gambling and tobacco use (4/7 articles), gambling and alcohol use (7/8 articles), and gambling and illicit drug use (7/8 articles). Conclusions The wide range in rates of gambling and problem gambling may be due in part to differences among the studies in participant samples, sampling techniques, assessment time frames, and definitions of gambling. Despite methodological differences, most studies showed significant associations of gambling with tobacco, alcohol, and other illicit drug use. Scientific Significance As accessibility to gambling increases, more research is needed to inform prevention efforts and identify youth at-risk for gambling and other high-risk behaviors. (Am J Addict 2015;24:206–216)

JWH-018 is a synthetic CB1 and CB2 agonist illegally marketed as products named “Spice” or “herbal blend” for its psychoactive effects which are much higher than those produced by cannabis. In the last year, the European Monitoring Centre for Drugs and Drug Addiction reported to the Italian National Early Warning System the seizure of plant material containing new halogenated derivatives of JWH-018 (JWH-018 Cl and JWH-018 Br). The present study aimed to investigate the in vitro and in vivo activity of these two new synthetic cannabinoids in mice. In vitro competition binding experiments performed on mouse and human CB1 receptors revealed a high affinity and potency of the halogenated compounds. Synthetic cannabinoids (0.01–6 mg/kg i.p.) impaired motor activity and induced catalepsy in mice and their effects were more severe with respect to those evoked by Δ9-THC. Moreover, they increased the mechanical and thermal pain threshold and induced a marked hypothermia. It is interesting to note that whereas high doses of JWH-018 cause seizures, myoclonia and hyperreflexia, the halogenated compounds, in particular JWH-018Br, were less effective. Behavioral and neurological changes were prevented by the selective CB1 receptor antagonist AM 251. These data demonstrate for the first time that JWH-018 Cl and JWH-018 Br act similarly to JWH-018 while inducing less convulsive episodes and myoclonias. These data support the hypothesis that the halogenated compounds may have been introduced onto market to produce similar intoxicating effects as JWH-018 while causing less side effects.

The paper describes a liquid chromatography/high resolution mass spectrometry LC/HRMS method for the simultaneous identification and quantification of stimulants (ephedrines, caffeine, anorectic drugs such as phentermine, phendimetrazine, phenmetrazine, fenfluramine, benfluorex, mephentermine, fencanfamine, sibutramine) and PDE5I (sildenafil, vardenafil and tadalafil) in food supplements using a benchtop Orbitrap mass spectrometer. The mass detector, with a nominal resolving power of 100,000 (FWHM at m / z 200), operated in full scan mode in ESI positive ionization mode. Analytes were identified by retention times, accurate masses and correspondence of experimental and calculated isotopic patterns. The limits of detection (LOD) obtained varied from 1 to 25 ng g −1 and limits of quantification (LOQ) were 50 ng g −1 for all compounds. The method was linear for all the analytes in the ranges from 50 to 2000 ng g −1 , giving correlation coefficients > 0.99. Accuracy (intended as %E) and repeatability (% CV) were always lower than 15%. The method was applied to the analysis of 36 dietary supplements, revealing the presence of ephedrine and/or pseudoephedrine in four of them, caffeine in eight of them and sildenafil in four of them. In one case, ephedrine was not reported on the label of the dietary supplement, as well as for caffeine in other two cases. A further confirmation of the analytes identity in positive samples was obtained through in-source fragmentation and comparison of the obtained fragments and their relative abundances with those from certified standards. As the acquisition mode is full scan, it would be also possible to re-process a previously acquired datafile for the investigation of untargeted analytes.