Genetic variation of the dopamine transporter (DAT1) influences the acute subjective responses to cocaine in volunteers with cocaine use disorders.

OBJECTIVE: The aim of this study was to identify gene variants of DAT1 (SLC6A3) that modulate subjective responses to acute cocaine exposure. METHODS: Non-treatment-seeking volunteers (n=66) with cocaine use disorders received a single bolus infusion of saline and cocaine (40 mg, intravenous) in a randomized order. Subjective effects were assessed with visual analog scales administered before (-15 min) and up to 20 min after infusion. Ratings of subjective effects were normalized to baseline, and saline infusion values were subtracted. Data were analyzed using repeated measures analysis of variance. DNA from the participants was genotyped for the DAT1 intron 8 (rs3836790) and 3'-untranslated region (rs28363170) variable number of tandem repeats. RESULTS: Participants were mostly male (∼80%) and African American (∼70%). No differences were found among drug use variables between groups for either polymorphism. Carriers of the 9-allele of the DAT1 3'-untranslated region (9,9 and 9,10) exhibited greater responses to cocaine for 'high', 'any drug effect', 'anxious', and 'stimulated' (all P-values<0.001) compared with individuals homozygous for the 10-allele. For the intron 8 polymorphism, individuals homozygous for the 6-allele exhibited greater responses for 'anxious' compared with carriers of the 5-allele (P<0.001). Individuals possessing the genotype pattern of 10,10 and at least one 5-allele reported lower responses to 'good effects', 'bad effects', 'depressed', and 'anxious' (all P-values<0.01). CONCLUSION: The data presented here show for the first time support for the hypothesis that genetic differences in DAT1 contribute to the variation in subjective responses to cocaine among participants with cocaine use disorders.

Normalized diurnal and between-day trends in illicit and legal drug loads that account for changes in population.

Drug concentrations in composite municipal wastewater samples and census-based estimates of population are used to derive daily loads of illicit substances that are indexed to population. However, such estimates do not provide information on the diurnal trends of substance excretion nor can they account for changes in population. To address these limitations, a series of 1 h composites created by sampling wastewater influent at 6 min intervals was collected over four consecutive days at a single wastewater treatment plant. Creatinine (a urinary indicator), caffeine, methamphetamine, benzoylecgonine (BZE), and cocaine were analyzed by liquid chromatography/tandem mass spectrometry (LC-MS/MS). Diurnal trends and between-day trends were substance specific and related to the number of estimated doses and excretory half-life. Normalization to creatinine yielded trends in substances that differed significantly from non-normalized trends by accounting for changes in population within the municipality studied. Increases in normalized substance excretion observed during early morning hours originate from individuals among the resident population of the municipality due to the absence of commuters.

Analysis of androgenic steroids in environmental waters by large-volume injection liquid chromatography tandem mass spectrometry.

A new method was developed for the analysis of natural and synthetic androgenic steroids and their selected metabolites in aquatic environmental matrixes using direct large-volume injection (LVI) high-performance liquid chromatography (HPLC) tandem mass spectrometry (MS/MS). Method accuracy ranged from 87.6 to 108% for analytes with well-matched internal standards. Precision, quantified by relative standard deviation (RSD), was less than 12%. Detection limits for the method ranged from 1.2 to 360 ng/L. The method was demonstrated on a series of 1 h composite wastewater influent samples collected over a day with the purpose of assessing temporal profiles of androgen loads in wastewater. Testosterone, androstenedione, boldenone, and nandrolone were detected in the sample series at concentrations up to 290 ng/L and loads up to 535 mg/h. Boldenone, a synthetic androgen, had a temporal profile that was strongly correlated to testosterone, a natural human androgen, suggesting its source may be endogenous. An analysis of the sample particulate fraction revealed detectable amounts of sorbed testosterone and androstenedione. Androstenedione sorbed to the particulate fraction accounted for an estimated 5 to 7% of the total androstenedione mass.

Wastewater testing compared with random urinalyses for the surveillance of illicit drug use in prisons.

INTRODUCTION AND AIMS: Illicit drug use is known to occur among inmate populations of correctional (prison) facilities. Conventional approaches to monitor illicit drug use in prisons include random urinalyses (RUA). Conventional approaches are expected to be prone to bias because prisoners may be aware of which days of the week RUAs are conducted. Therefore, we wanted to compare wastewater loads for methamphetamine and cocaine during days with RUA testing and without. DESIGN AND METHODS: We collected daily 24-h composite samples of wastewater by continuous sampling, computed daily loads for 1 month and compared the frequency of illicit drug detection to the number of positive RUAs. Diurnal data also were collected for 3 days to determine within-day patterns of illicit drugs excretion. RESULTS: Methamphetamine was observed in each sample of prison wastewater with no significant difference in daily mass loads between RUA testing and non-testing days. Cocaine and its major metabolite, benzoylecgonine, were observed only at levels below quantification in prison wastewater. Six RUAs were positive for methamphetamine during the month while none were positive for cocaine out of the 243 RUAs conducted. DISCUSSION AND CONCLUSIONS: Wastewater analyses offer data regarding the frequency of illicit drug excretion inside the prison that RUAs alone could not detect.

Using wastewater-based epidemiology to estimate drug consumption-Statistical analyses and data presentation.

AIM: Analysis of wastewater samples can be used to assess population drug use, but reporting and statistical issues have limited the utility of the approach for epidemiology due to analytical results that are below the limit of quantification or detection. Unobserved or non-quantifiable-censored-data are common and likely to persist as the methodology is applied to more municipalities and a broader array of substances. We demonstrate the use of censored data techniques and account for measurement errors to explore distributions and annual estimates of the daily mean level of drugs excreted per capita. MEASUREMENTS: Daily 24-hour composite wastewater samples for 56days in 2009 were obtained using a random sample stratified by day of week and season for 19 municipalities in the Northwest region of the U.S. METHODS: Methamphetamine, benzoylecgonine (cocaine metabolite), 3,4 methylenedioxymethamphetamine (MDMA), methadone, oxycodone and hydrocodone were identified and quantified in wastewater samples. Four statistical approaches (reporting censoring, Maximum Likelihood Estimation, Kaplan-Meier estimates, or complete data calculations) were used to estimate an annual average, including confidence bounds where appropriate, dependent upon the amount of censoring in the data. FINDINGS: The proportion of days within a year with censored data varied greatly by drug across the 19 municipalities, with MDMA varying the most (4% to 94% of observations censored). The different statistical approaches each needed to be used given the levels of censoring of measured drug concentrations. Figures incorporating confidence bounds allow visualization of the data that facilitates appropriate comparisons across municipalities. CONCLUSIONS: Results from wastewater sampling that are below detection or quantification limits contain important information and can be incorporated to create a more complete and valid estimate of drug excretion.

Analysis of Nucleosides in Municipal Wastewater by Large-Volume Liquid Chromatography Tandem Mass Spectrometry.

Nucleosides are components of both DNA and RNA, and contain either a ribose (RNA) or 2deoxyribose (DNA) sugar and a purine or pyrimidine base. In addition to DNA and RNA turnover, modified nucleosides found in urine have been correlated to a diminished health status associated with AIDS, cancers, oxidative stress and age. Nucleosides found in municipal wastewater influent are potentially useful markers of community health status, and as of now, remain uninvestigated. A method was developed to quantify nucleosides in municipal wastewater using large-volume injection, liquid chromatography, and mass spectrometry. Method accuracy ranged from 92 to 139% when quantified by using isotopically labeled internal standards. Precision ranged from 6.1 to 19% of the relative standard deviation. The method's utility was demonstrated by the analysis of twenty-four hour composite wastewater influent samples that were collected over a week to investigate community nucleoside excretion. Nucleosides originating from RNA were more abundant that DNA over the study period, with total loads of nucleosides ranging from 2 to 25 kg/day. Given this relatively high amount of nucleosides found over the study period they present an attractive analyte for the investigation of community health.

The influence of smoking cigarettes on the high and desire for cocaine among active cocaine users.

The primary aim was to evaluate whether nicotine use alters the high or desire for cocaine among active cocaine users who concurrently smoke cigarettes. Participants answered the Fagerstrom Test for Nicotine Dependence (FTND), Nicotine-Stimulant Interaction Questionnaire (NSIQ), and Multiple Drug Use Questionnaire (MDUQ). These questionnaires employ subject recall of participants' drug use habits. The participants that smoked (N=163/188) were primarily African American males who were 45.0±0.5 (mean±S.E.M.) years of age, and used cocaine for 17.9±0.6 years and 19.8±0.6 days out of the last 30. These individuals smoked 14.0±0.8 cigarettes/day (CPD), scored 4.6±0.2 (on a scale of 0-10) on the FTND, and smoked cigarettes for 23.5±0.7 years. Two questions from the MDUQ, which evaluates the interaction between cocaine and nicotine, (-5: reduces effect, 0: no change, +5: increases effect) included "Does nicotine affect the high that you experience from cocaine?" and "Does nicotine affect your desire for cocaine?", and the scores were 1.3±0.2 and 0.8±0.2, respectively. The NSIQ also evaluated interactive effects of nicotine and cocaine, on a scale of 0 to 100 (0: not at all, 100: most ever). Smokers responded most strongly that using cocaine increased both the urge to smoke and cigarette craving. Additional analyses were performed by separating participants into HighCPD vs. LowCPD groups via median split. The HighCPD group smoked 22.7±1.1 CPD while the LowCPD group smoked 6.4±0.3 CPD [F(1,161)=228.4, p<0.0001], and the HighCPD group had a mean FTND score twice that of the LowCPD group. Significant differences emerged between the two groups on multiple items of the NSIQ, but not the MDUQ. The subjective ratings of high and desire for cocaine, and several subjective effects produced by cocaine, were modestly altered by cigarette smoking. Taken together, these data suggest that cigarette smoking may augment the craving and high produced by cocaine.