Determination of cortisol in hair using UHPLC–MS/MS: application to patients admitted for ethanol dependence treatment

Aim: Cortisol hair levels can be used to evaluate chronic stress status. In this context, an improved UHPLC–MS/MS assay for the determination of cortisol in hair was developed and validated. Materials & methods: Hair was extracted with methanol for 4 h at 25°C. Chromatographic run time was 5.5 min. The assay was linear in the range of 1–250 pg mg-1. Precision was 3.6–12.2% and accuracy 97.1–103.8%. The method was applied in hair from 19 volunteers admitted at a rehabilitation clinic, with ethanol consumption classified using ethyl glucuronide hair levels. Conclusion: Abstinent/chronic moderate ethanol consumers had significantly lower cortisol hair levels than chronic excessive consumers. This is the first study evaluating cortisol hair levels in ethanol abuse patients using an objective marker for ethanol consumption.

SPECIFICS OF ALCOHOL USE DISORDER IN PRE-HOSPITAL PRACTICES AND EMERGENCY UNIT

Ethanol intoxication is analyzed from the point of ethanol interaction with receptors and enzymes. Ethanol dependence is based on neurochemical misbalance, which is different in acute ethanol intoxication and withdrawal. It is shown that the clinical manifestation of ethanol intoxication and its complications depends on ligand-receptor and enzyme imbalances. Pharmacological approaches to the treatment of acute ethanol intoxication and its complications at the pre-hospital stage and in the emergency, unit are discussed.

Evaluation of anxiety-like behaviors following ethanol withdrawal in mice: effects of cannabidiol

The abrupt interruption of ethanol consumption increases anxiety-like behaviors in rodents and may reflect different aspects of ethanol dependence in humans. Measuring emotional behaviors resulting from ethanol withdrawal may aid in testing potential pharmacological agents for the treatment of ethanol dependence. In the present study, we used forced expositon to ethanol 20% during 10 days to mice, followed by abrupt withdrawal of the substance. The animals were evaluated 7, 24 and 35 h after ethanol withdrawal in three different behavioral paradigms, i.e., the open field (OF), light dark (LD) transition and elevated plus maze (EPM), tests usually used to measure anxiety-like behaviors. This was done with the aim of identifying the best interval as well as the most appropriate behavioral test to detect the effects of drugs that can relieve anxiety induced by ethanol withdrawal in mice. We also evaluated the effect of cannabidiol (CBD 10, 30 and 60 mg/kg) in ethanol withdrawn mice because it has been shown to alliviate drug addicton and present anti-anxiety effects. Our results show significant behavioral changes at 24 h following ethanol withdrawal. Diazepam (4 mg/kg), used as a positive control, counteracted the effects of ethanol withdrawal in OF, LD box and EPM. Cannabidiol attenuated anxiety-like behavior produced at 24 h after abstinence from ethanol exposure only in the EPM test.

Inhibition of the Ventral Hippocampus Increases Alcohol Drinking

The function of the ventral hippocampus (vHC) supports many behaviors, including those related to reward seeking behaviors and drug addiction. We used a mouse model of alcohol dependence and relapse to investigate the role of the vHC in alcohol (ethanol) drinking. One experiment used a chemogenetic approach to inhibit vHC function while ethanol dependent and non-dependent mice had access to ethanol. Interestingly, the non-dependent mice expressing an inhibitory DREADD in the VHC showed a significant increase in ethanol drinking (∼30%) after hM4Di DREADD activation with clozapine-n-oxide (CNO; 3 mg/kg, ip.) compared to vehicle. On the other hand, ethanol dependent mice, which were already drinking significantly more ethanol than non-dependent mice, only had a slight, non-significant increase in drinking after CNO challenge. In a separate group of dependent and non-dependent mice, GCaMP6f calcium-dependent activity was recorded in the vHC while mice were actively drinking ethanol. These data showed that once mice were rendered ethanol dependent and were drinking more ethanol than the non-dependent mice, calcium signaling in the ventral hippocampus decreased (∼45%) in the ethanol dependent mice compared to the non-dependent mice. Together, these findings suggest that ethanol dependence reduces activity of the ventral hippocampus and that reduced function of this brain region contributes to increased ethanol drinking.