Carrier-mediated cocaine transport at the blood-brain barrier as a putative mechanism in addiction liability.

BACKGROUND: The rate of entry of cocaine into the brain is a critical factor that influences neuronal plasticity and the development of cocaine addiction. Until now, passive diffusion has been considered the unique mechanism known by which cocaine crosses the blood-brain barrier. METHODS: We reassessed mechanisms of transport of cocaine at the blood-brain barrier using a human cerebral capillary endothelial cell line (hCMEC/D3) and in situ mouse carotid perfusion. RESULTS: Both in vivo and in vitro cocaine transport studies demonstrated the coexistence of a carrier-mediated process with passive diffusion. At pharmacological exposure level, passive diffusion of cocaine accounted for only 22.5% of the total cocaine influx in mice and 5.9% in hCMEC/D3 cells, whereas the carrier-mediated influx rate was 3.4 times greater than its passive diffusion rate in vivo. The functional identification of this carrier-mediated transport demonstrated the involvement of a proton antiporter that shared the properties of the previously characterized clonidine and nicotine transporter. The functionnal characterization suggests that the solute carrier (SLC) transporters Oct (Slc22a1-3), Mate (Slc47a1) and Octn (Slc22a4-5) are not involved in the cocaine transport in vivo and in vitro. Diphenhydramine, heroin, tramadol, cocaethylene, and norcocaine all strongly inhibited cocaine transport, unlike benzoylecgonine. Trans-stimulation studies indicated that diphenhydramine, nicotine, 3,4-methylenedioxyamphetamine (ecstasy) and the cathinone compound 3,4-methylenedioxypyrovalerone (MDPV) were also substrates of the cocaine transporter. CONCLUSIONS: Cocaine transport at the BBB involves a proton-antiporter flux that is quantitatively much more important than its passive diffusion. The molecular identification and characterization of this transporter will provide new tools to understand its role in addictive mechanisms.

Simultaneous determination of cotinine and trans-3-hydroxycotinine in urine by automated solid-phase extraction using gas chromatography-mass spectrometry.

A gas chromatography-mass spectrometry method was developed and validated for the simultaneous automated solid-phase extraction and quantification of cotinine and trans-3-hydroxycotinine in human urine. Good linearity was observed over the concentration ranges studied (R(2) > 0.99). The limit of quantification was 10 ng/mL for both analytes. The limits of detection were 0.06 ng/mL for cotinine (COT) and 0.02 ng/mL for trans-3-hydroxycotinine (OH-COT). Accuracy for COT ranged from 0.98 to 5.28% and the precision ranged from 1.24 to 8.78%. Accuracy for OH-COT ranged from -2.66 to 3.72% and the precision ranged from 3.15 to 7.07%. Mean recoveries for cotinine and trans-3-hydroxycotinine ranged from 77.7 to 89.1%, and from 75.4 to 90.2%, respectively. This analytical method for the simultaneous measurement of cotinine and trans-3-hydroxycotinine in urine will be used to monitor tobacco smoking in pregnant women and will permit the usefulness of trans-3-hydroxycotinine as a specific biomarker of tobacco exposure to be determined.

Determination of levamisole in the plasma of patients with falciparum malaria using high-performance liquid chromatography.

BACKGROUND: The oral antihelmintic drug levamisole reduces sequestration of late stage parasites in falciparum malaria. Levamisole has been also identified as a cocaine adulterant. In the present study, authors developed a sensitive and selective HPLC-assay for the determination of levamisole in the plasma from patients with falciparum malaria. METHODS: Chromatographic separation was achieved by using a C18 column and with an isocratic elution system comprising phosphate buffer and acetonitrile. The eluate was monitored at 235 nm by diode array detection. RESULTS: The calibration curve for levamisole was linear in the range from 50 to 2000 ng/mL (r2 > 0.999). The limit of quantification was 28 ng/mL and the inter- and intraday coefficients of variation were less than 7%. No interference from commonly prescribed malaria treatments was observed. CONCLUSIONS: The HPLC method is simple, rapid, and robust and is suited for monitoring levamisole patients in routine or toxicological studies.

Respiratory toxicity of buprenorphine results from the blockage of P-glycoprotein-mediated efflux of norbuprenorphine at the blood-brain barrier in mice.

OBJECTIVES: Deaths due to asphyxia as well as following acute poisoning with severe respiratory depression have been attributed to buprenorphine in opioid abusers. However, in human and animal studies, buprenorphine exhibited ceiling respiratory effects, whereas its metabolite, norbuprenorphine, was assessed as being a potent respiratory depressor in rodents. Recently, norbuprenorphine, in contrast to buprenorphine, was shown in vitro to be a substrate of human P-glycoprotein, a drug-transporter involved in all steps of pharmacokinetics including transport at the blood-brain barrier. Our objectives were to assess P-glycoprotein involvement in norbuprenorphine transport in vivo and study its role in the modulation of buprenorphine-related respiratory effects in mice. SETTING: University-affiliated research laboratory, INSERM U705, Paris, France. SUBJECTS: Wild-type and P-glycoprotein knockout female Friend virus B-type mice. INTERVENTIONS: Respiratory effects were studied using plethysmography and the P-glycoprotein role at the blood-brain barrier using in situ brain perfusion. MEASUREMENTS AND MAIN RESULTS: Norbuprenorphine(≥ 1 mg/kg) and to a lesser extent buprenorphine (≥ 10 mg/kg) were responsible for dose-dependent respiratory depression combining increased inspiratory (TI) and expiratory times (TE). PSC833, a powerful P-glycoprotein inhibitor, significantly enhanced buprenorphine-related effects on TI (p < .01) and TE (p < .05) and norbuprenorphine-related effects on minute volume (VE, p < .05), TI, and TE (p < .001). In P-glycoprotein-knockout mice, buprenorphine-related effects on VE (p < .01), TE (p < .001), and TI (p < .05) and norbuprenorphine-related effects on VE (p < .05) and TI (p < .001) were significantly enhanced. Plasma norbuprenorphine concentrations were significantly increased in PSC833-treated mice (p < .001), supporting a P-glycoprotein role in norbuprenorphine pharmacokinetics. Brain norbuprenorphine efflux was significantly reduced in PSC833-treated and P-glycoprotein-knockout mice (p < .001), supporting P-glycoprotein-mediated norbuprenorphine transport at the blood-brain barrier. CONCLUSIONS: P-glycoprotein plays a key-protective role in buprenorphine-related respiratory effects, by allowing norbuprenorphine efflux at the blood-brain barrier. Our findings suggest a major role for drug-drug interactions that lead to P-glycoprotein inhibition in buprenorphine-associated fatalities and respiratory depression.

Stability of levamisole oral solutions prepared from tablets and powder.

PURPOSE: To study the stability of levamisole oral solutions (25 mg/mL) prepared from powder and tablets stored at 4 +/- 3 degrees C and 23 +/- 2 degrees C in amber glass prescription bottles. METHODS: Levamisole 25 mg/mL solutions were prepared from commercially available 50-mg tablets or from pure powder in sterile water. Levamisole concentrations were determined in duplicate by a stability-indicating HPLC method at 0, 1, 2, 3, 4, 7, 14, 30, 60 and 90 days. The initial and final pHs of solutions were measured. RESULTS: The recovery of levamisole from tablets was 100 +/- 2.1%. No color or odour changes were observed during the study period. The oral solutions prepared from powder were stable at least 90 days stored at 4 and 23 degrees C. The oral solutions prepared from tablets were stable at least 90 days at 4 degrees C and 15 days when stored at 23 degrees C. The initial pH of solutions prepared from powder and tablets were 5.30 and 4.55, respectively. Initial and final pH values were significantly different (p<0.001) for the two solutions. CONCLUSIONS: Levamisole 25 mg/mL oral solutions can be prepared from tablets or powder with sterile water for irrigation and stored for 90 days under refrigeration, taking account of the lack of microbiological contamination.

Determination and Validation of a Solid-phase Extraction Gas Chromatography-mass Spectrometry for the Quantification of Methadone and Its Principal Metabolite in Human Plasma.

This study aimed to develop a solid-phase extraction gas chromatography-selected ion monitoring-mass spectrometry method for the determination of methadone (MDN) and 2-ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine (EDDP) in human plasma. The linear response was obtained over the concentration range from 10 to 2000 ng/mL for MDN and EDDP. The absolute recoveries of MDN and EDDP were 95.9%-98.9% and 94.8%-102.4%, with relative standard deviation (RSD) ranging from 1.8% to 2.7% and 1.8% to 3.9%, respectively. The intra- and interday precisions were found to be less than 5% for both analytes. The limits of detection of MDN and EDDP were 4 and 5 ng/mL, respectively. The presented method was convenient for therapeutic drug monitoring and pharmacokinetic studies in patients on heroin-assisted MDN therapy.

Stability of sulfadiazine oral liquids prepared from tablets and powder.

PURPOSE: To assess the stability of sulfadiazine (SDZ) oral liquids prepared from tablets and powder at two temperatures. METHODS: Solutions of SDZ 200 mg/mL were prepared from commercially available 500 mg tablets and powder in sterile water for irrigation. They were stored in amber glass bottles at 4 degrees C and 23 degrees C. The concentrations of SDZ were determined in duplicate by high-performance liquid chromatography at 0, 1, 3, 7 and 14 days. The initial and final pH of solutions was compared. The recovery of SDZ from tablets was determined. A loss exceeding 10% of the initial concentration of SDZ was considered excessive degradation. RESULTS: The recovery of SDZ from tablets was 100 +/- 3%. The initial pH values were significantly different between solutions prepared from tablets and powder, 6.9 and 9.8 respectively. No significant difference was found between initial and final pH values for the two all formulations. Detectable change in odor was observed for the solutions stored at 23 degrees C. The solution prepared from powder was stable 3 days stored at 4 degrees C. Other formulations lost over 10% of the initial SDZ concentration within 2 days. CONCLUSIONS: SDZ 200 mg/mL oral solution prepared from powder could be used to facilitate drug administration to very young children by nurses but by taking account of its fast degradation.

Buprenorphine and norbuprenorphine determination in mice plasma and brain by gas chromatography-mass spectrometry.

A gas chromatography tandem mass spectrometry method for quantification of buprenorphine (BUP) and norbuprenorphine (NBUP) in brain and plasma samples from mice was developed and validated. Analytes were extracted from the brain or plasma by solid phase extraction and quantified within 20 minutes. Calibration was achieved by linear regression with a 1/x weighting factor and d4-buprenorphine internal standard. All products were linear from 1 to 2000 ng/mL with a correlation of determination >0.99. Assay accuracy and precision of back-calculated standards were within ±10%. The lower limit of quantification for both BUP and NBUP from the brain and plasma was 1 ng/mL. This sensitive and specific method can be used for the investigation of BUP mechanism of action and clinical profile.

Impact of P-glycoprotein at the blood-brain barrier on the uptake of heroin and its main metabolites: behavioral effects and consequences on the transcriptional responses and reinforcing properties.

RATIONALE: Transport across the BBB is a determinant of the rate and extent of drug distribution in the brain. Heroin exerts its effects through its principal metabolites 6-monoacetyl-morphine (6-MAM) and morphine. Morphine is a known substrate of P-glycoprotein (P-gp) at the blood-brain-barrier (BBB) however, little is known about the interaction of heroin and 6-MAM with P-gp. OBJECTIVE: The objective of this paper is to study the role of the P-gp-mediated efflux at the BBB in the behavioral and molecular effects of heroin and morphine. METHODS: The transport rates of heroin and its main metabolites, at the BBB, were measured in mice by in situ brain perfusion. We then examined the effect of inhibition of P-gp on the acute nociception, locomotor activity, and gene expression modulations induced by heroin and morphine. The effect of P-gp inhibition during the acquisition of morphine-induced place preference was also studied. RESULTS: Inhibition of P-gp significantly increased the uptake of morphine but not that of heroin nor 6-MAM. Inhibition of P-gp significantly increased morphine-induced acute analgesia and locomotor activity but did not affect the behavioral effects of heroin; in addition, acute transcriptional responses to morphine were selectively modulated in the nucleus accumbens. Increasing morphine uptake by the brain significantly increased its reinforcing properties in the place preference paradigm. CONCLUSIONS: The present study demonstrated that acute inhibition of P-gp not only modulates morphine-induced behavioral effects but also its transcriptional effects and reinforcing properties. This suggests that, in the case of morphine, transport across the BBB is critical for the development of dependence.

A Rapid Stability-Indicating RP-HPLC Method for the Determination of Betaxolol Hydrochloride in Pharmaceutical Tablets.

A stability-indicating reversed-phase high performance liquid chromatography (RP-HPLC) method was developed for the determination of betaxolol hydrochloride, a drug used in the treatment of hypertension and glaucoma. The desired chromatographic separation was achieved on a Nucleosil C18, 4 µm (150 × 4.6 mm) column, using isocratic elution at a 220 nm detector wavelength. The optimized mobile phase consisted of a 0.02 M potassium dihydrogen phosphate: methanol (40:60, v/v, pH 3.0 adjusted with o- phosphoric acid) as solvent. The flow rate was 1.6 mL/min and the retention time of betaxolol hydrochloride was 1.72 min. The linearity for betaxolol hydrochloride was in the range of 25 to 200 µg/mL. Recovery for betaxolol hydrochloride was calculated as 100.01%-101.35%. The stability-indicating capability was established by forced degradation experiments and the separation of unknown degradation products. The developed RP-HPLC method was validated according to the International Conference on Harmonization (ICH) guidelines. This validated method was applied for the estimation of betaxolol hydrochloride in commercially available tablets.

Police custody following driving under the influence of cannabis: a prospective study.

Traffic offences are a common cause of detention in police custody. We hypothesized that drug intoxication while driving could correspond to specific medical conditions of the detainees. Our objective was to evaluate medical features and addictive behaviours of suspected drug drivers and to collect data regarding assaults or injuries in these individuals. We conducted a prospective study (April 2010-December 2011) of suspected drug driving arrestees, who were compared to drink drivers or persons aged over 18 detained for other reasons. Data collected concerned persons' characteristics, reported assaults, and observed injuries. A total of 205 drivers were tested positive for drugs in blood, 116 were either positive for drugs in urine or saliva and negative in blood, or negative in urine. Cannabis-only users accounted for 201 of 205 drug drivers (98%). Suspected drug driving arrestees had good overall health rating. Drug drivers were younger than controls and requested more rarely medical examination (12% vs. 44%, P<0.0001). They were rarely involved in addiction treatment (3%) and reported assaults or presented traumatic injuries less often than drink drivers and controls (8% vs. 38% and 25%, P<0.0001). Drug drivers were less often alcohol abusers than controls. Their opinion on custody was better than that of controls and they were considered unconditionally fit for detention more frequently (99% vs. 77%, P<0.0001). We conclude that arrested drug drivers were young, healthy, and infrequently reported assaults or presented traumatic injuries, which does not put them in a high risk medical condition. Medical care could include brief interventions on addictive behaviours.

Gender and strain contributions to the variability of buprenorphine-related respiratory toxicity in mice.

While most deaths from asphyxia related to buprenorphine (BUP) overdose have been reported in males, higher plasma concentrations of BUP and its toxic metabolite norbuprenorphine (NBUP) have been observed in females. We previously demonstrated that P-glycoprotein (P-gp) modulation at the blood-brain barrier (BBB) contributes highly to BUP-related respiratory toxicity, by limiting NBUP entrance into the brain. In this work, we sought to investigate the role of P-gp-mediated transport at the BBB in gender and strain-related variability of BUP and NBUP-induced respiratory effects in mice. Ventilation was studied using plethysmography, P-gp expression using western blot, and transport at the BBB using in situ cerebral perfusion. In male Fvb and Swiss mice, BUP was responsible for ceiling respiratory effects. NBUP-related reduction in minute volume was dose-dependent but more marked in Fvb (p<0.01 at 1mg/kg NBUP and p<0.001 at 3 and 9mg/kg NBUP) than in Swiss mice (p<0.001 at 9mg/kg NBUP). Female Fvb mice were more susceptible to BUP than males with significantly increased inspiratory time (p<0.05) and to NBUP with significantly increased expiratory time (p<0.01). Following BUP administration, plasma BUP concentrations were significantly higher (p<0.01) and plasma NBUP concentrations significantly lower (p<0.001) in Fvb mice compared to Swiss mice. Plasma BUP concentrations were significantly higher (p<0.05) and plasma NBUP concentrations significantly lower (p<0.01) in male compared to female Fvb mice. In contrast, following NBUP administration, comparable plasma NBUP concentrations were observed in both genders and strains. No differences in P-gp expression or BUP and NBUP transport across the BBB were observed between male and female Fvb mice as well as between Swiss and Fvb mice. Our results suggest that P-gp-mediated transport across the BBB does not play a key-role in gender and strain-related variability in BUP and NBUP-induced respiratory toxicity in mice. Both gender- and strain-related differences in respiratory effects of BUP could be attributed to BUP itself rather than to its metabolite, NBUP.

[Spice in France: mixed herbs containing synthetic cannabinoids]. / Le spice en France: mélange d'herbes contenant des cannabinoïdes de synthèse.

Smokable herbal mixtures under the brand name Spice were first sold on the Internet and in various specialised shops in 2006 or earlier. When smoked, the Spice products have effects similar to those of cannabis. Forensic investigations were undertaken by German and Austrian authorities in order to identify the psychoactive ingredients of Spice. A new psychoactive substance JWH-018 has been identified in Spice products. JWH-018 was first synthesized in 1995 and produces effects similar to those THC. Subsequently, the synthetic cannabinoid CP 47,497 was also identified. Outside of Europe, the United States Drug Enforcement Administration reported that another potent synthetic cannabinoid, HU-210, had been found. Since 2009, other synthetic cannabinoids were identified in Europe. None of the above-mentioned synthetic cannabinoids is internationally controlled as a drug and there is no information on any of them having been authorised as a medicinal product in the European Union. There are no officially published safety data and little is known about their effects in humans. Some of the characteristics of these compounds, e.g. volatility and activity in small doses, are likely to present further analytical and toxicological challenges. Responding to potential health concerns, Europe has taken legal actions to ban or otherwise control Spice products and related compounds.

Police custody following drink-driving: a prospective study.

BACKGROUND: Drink-driving is a crime and traffic offences are a common cause of detention in police custody. Legal assessment of alcohol intoxication is based on breath or blood testing. We hypothesize that refusal of breath alcohol testing or inability to perform it can correspond to singular medical characteristics of the detainee, possibly assaulted or injured during the arrest. Our objective was to determine medical characteristics of detainees held in custody for drink-driving. METHODS: Prospective monocentric study (April-October, 2010) of drink-drive arrestees. Controls were persons aged over 18 detained for other reasons than drink-driving. Data collected concerned persons' characteristics and reported assaults or observed injuries. RESULTS: 223 drivers were tested positive for breath alcohol level and 55 suspected drink-drivers refused or were not able to complete breath test. 2212 consecutively examined persons served as controls. Drink-drive arrestees requested medical examination more rarely (18% and 7%, vs. 43%, P<0.0001) and drivers tested positive for breath alcohol were more frequently alcohol abusers (25% vs. 14%, P<0.0001) than controls. Drivers who did not complete breath test more often reported assaults than those tested positive for breath alcohol (22% vs. 8%, P=0.007). They had more frequent traumatic injuries than those tested positive and than controls (29% vs. 11% and 17%, P=0.003 and 0.02). Only 1% of drink drivers were unfit for detention after medical examination. CONCLUSION: Physicians need to give attentive care to detained drink-drivers. Special attention should be paid to drink-drivers who refused or were not able to complete breath alcohol measurement.

[Crystal meth: a particular form of methamphetamine]. / Crystal meth : une forme de méthamphétamine.

UNLABELLED: Crystal Meth: is a synthesis drug whose consumption developed with the beginning of the year 2000 in Europe. Crystal can be swallowed, crunched, smoked, injected or inserted by rectal way. Required effects: "crystal" removes tiredness, brings a feeling of power and of self-control, makes trustful, sexy and merry. Clinical effects: by its sympathomimetic action-like, "crystal" causes hypertension and tachycardia. It crosses the placenta and and is excreted in the mother's milk. It brings to a dependence similar to that of cocaine. COMPLICATIONS: "crystal" can generate cardiopulmonary complications, can reveal cognitive and psychological disorders. It leads the consumers to have not protected and repeated sexual relations. CONCLUSION: in France, the consumption of "crystal" remains marginal. Plan 2007-2011 of assumption of responsibility and prevention of the addictions integrates the elements such as the prevention to fight against the extension of this drug.

[Misoprostol for treating postpartum hemorrhages]. / Le misoprostol dans le traitement curatif des hémorragies du post-partum.

Postpartum hemorrhage is defined by bleeding > 500 mL through the vagina. It is one of the obstetrical complications that obstetricians fear most. It is the leading cause of maternal mortality in the world, especially in developing countries. The reference treatments in France are parenteral oxytocin and sulprostone. Sulprostone involves sometimes fatal side effects, and must be administered only in appropriate health care facilities. It also has the major disadvantage of requiring refrigeration. Misoprostol has uterotonic properties that have led to its occasional off-label use in the treatment of postpartum hemorrhage, by rectal or sublingual administration, as an alternative to sulprostone. A careful review of the literature on this particular use of misoprostol is essential.