Driving under the influence of drugs: Prevalence in road traffic accidents in Italy and considerations on per se limits legislation.

OBJECTIVE: The objective of this study was to present the prevalence and concentrations of drugs in blood samples of drivers involved in road traffic accidents (RTAs) and to discuss the effects of adopting different concentration cutoff values proposed or applied in other European countries on the number of driving under the influence of drugs (DUID) offenses. METHODS: Blood samples from drivers involved in RTAs in Padova province from 2014 to 2017 were analyzed for the presence of alcohol and drugs. The prevalence of positive subjects was reported for each substance adopting the limits of quantification (LOQs) of our laboratory and the concentration cutoff values proposed and/or used in other European countries. The reduction of cases of driving under the influence of illicit drugs in applying different cutoffs was calculated. RESULTS: Four thousand four hundred forty-three blood samples were analyzed: 23.7% were positive for alcohol and 19.9% for psychoactive drugs, with prevalences of polydrug and alcohol-drug abuse of 4.5 and 6%, respectively. The most frequently detected drugs were cannabinoids (9.7%) and cocaine (7.2%), followed by benzodiazepines (4.1%), opiates (1.9%), and other opioids (1.7%). Barbiturates, amphetamines, and ketamine were identified in a much smaller number of cases. The overall decrease in DUID cases when adopting different cutoffs with respect to cases above the LOQs was between 8 and 84%. The adoption of high LOQs such as those used in the European Union's research project on Driving Under the Influence of Drugs, Alcohol and Medicines (DRUID) decreases the hypothetical number of DUID offenses by a quarter, and per se limits proposed as broadly equivalent to a blood alcohol concentration (BAC) between 0.2 and 0.8 g/L dramatically reduce the cases of DUID (cocaine -81%, cannabis -79%, opioids -97%, opiates -96%, and amphetamines -77%); no ketamine-positive samples were above the cutoff. CONCLUSIONS: The implementation of high analytical limits or per se limits based on impairing concentrations in the Italian legislation could result in the prosecution of a much lower number of drugged drivers involved in RTAs, with a decrease from 25% to more than 80% depending on the limits.

Occupational exposure to ketamine detected by hair analysis: a retrospective and prospective toxicological study.

Ketamine (KT) is used to induce and maintain general anaesthesia in combination with sedative drugs in human and animals. Because of its dissociative and hallucinogenic effects, KT has become a recreational drug in a variety of social settings and may be included in the panel of drugs of abuse that are controlled in driving under the influence (DUI) ascertainment. In a local driving license re-granting protocol, a case where a veterinary physician was found positive to KT and nor-ketamine (NK) in hair suggested the possibility of a professional exposure in a veterinary setting and prompted an experimental study. Male (7) and female (4) veterinary physicians were recruited on a voluntary base. Detailed information was collected on their habits, use of drugs, professional practice, frequency and mode of using KT injections. Hands and skin were examined. Head hair and pubic hair were collected. Two naïve subjects, starting their professional practice at a local veterinary clinic, were recruited and their hair (head, pubic, axillary, thoracic hair, and beard) and urine were collected before and after usual clinic activity. Hair were cut according to their length, washed, pulverized and 25mg were extracted and analyzed by liquid chromatography coupled to high accuracy, high resolution mass spectrometry. All the hair samples from the veterinary physicians turned to be positive for KT, at a concentration varying from 0.010 to 0.840ng/mg in head hair and from 0.040 to 2.04ng/mg in pubic hair; NK ranged from not detected to 0.080ng/mg in head hair, from not detected to 0.100 in pubic hair; when KT was ≥0.100, NK was always detected. For the two naïve subjects, hair from different body sites were negative before they started their activity, and positive one month later; some urine samples resulted positive and confirmed systemic exposure to KT. The possibility of unaware exposure to KT was demonstrated. The site of absorption is skin, independently from the presence of skin injuries due to hand contact with KT injection solutions and/or animal body fluids by veterinary physicians during clinical activities and animal handling. Possible adverse systemic effects by unaware KT exposure need to be studied and concerns have arisen as to healthy and safe workplace policies. Comparison with a population of subjects undergoing driving license re-granting evidenced a larger range of concentration in this latter setting (0.050-10.0ng/mg for KT, not detected to 0.100ng/mg for NK).

A matrix-assisted laser desorption/ionization mass spectrometry study of the non-enzymatic glycation products of human globins in diabetes.

The molecular species present in globins from healthy and diabetic subjects with and without chronic complications have been analyzed by matrix-assisted laser desorption/ionization mass spectrometry. The technique demonstrated the presence of glycated and glyco-oxidated species of both alpha- and beta-globins. Their abundances show a good linear relationship with respect to HbA1c values and with the mean daily plasma glucose levels over the 6 weeks preceding the investigation. Interestingly, slightly different behaviour is observed in the data from patients with and without chronic complications; the plots of HbA1c vs. the abundance of glycated and glyco-oxidated species show different slopes and different intercepts with the y-axis. To investigate this aspect the mean abundances of glyco-oxidated species from healthy subjects and from diabetic patients with and without complications were calculated. Higher values were found for the two last sets of samples, but no significant difference was found between them. These data could indicate different individual proclivities to oxidation and/or different oxidation kinetics related to behavioural and environmental factors.

The complexity of non-enzymatic glycation product sets of human globins.

AIMS/HYPOTHESIS: Recently an individual variability in the relationships between mean blood glucose levels and HbA1c has been observed among diabetic patients. The aim of this study was to provide an accurate description and evaluation of glycated and glyco-oxidated globins from diabetic subjects and their relationship with HbA1c and plasma glucose values. METHODS: We studied 20 type 2 diabetic and 10 healthy subjects. Plasma samples were analysed by matrix-assisted laser desorption ionisation mass spectrometry. RESULTS: The presence of glycated and glyco-oxidated species of both alpha and beta globin was demonstrated. Values for these showed a good linear relationship with HbA1c values and the mean daily plasma glucose values for the 6 weeks preceding the investigation. Trends differed according to whether patients had chronic complications or not, differences being seen in the slopes of the plots relating HbA1c to the abundance of glycated and glyco-oxidated species. CONCLUSIONS/INTERPRETATION: The data obtained are consistent with the concept that individuals have a different individual proclivity for oxidation and/or that different oxidation kinetics are related to behavioural and environmental factors. Our data are thus relevant to the analysis of phenotype differences in diabetic patients.

Melanogenesis by tyrosinase action on 3,4-dihydroxyphenylalanine (DOPA) in the presence of polyethylene glycol: a matrix-assisted laser desorption/ionization mass spectrometric investigation.

The enzymatic reaction between DOPA and tyrosinase, the enzyme considered to be responsible for melanogenesis, was carried out in the presence of polyethylene glycol (PEG). This choice was made in order to increase the solubility of melanins, since these polymers are highly insoluble. The reaction mixtures were sampled at different times, immediately ultrafiltered to remove the enzyme, lyophilized, and analyzed by matrix-assisted laser desorption/ionization (MALDI) mass spectrometry. The results were very different from those obtained in the absence of PEG. Only a few oligomers of dihydroxyindole (DHI) and dihydroxyindole-2-carboxylic acid (DHICA) were detected in low abundances, whereas new species originating from reaction of PEG with species belonging to the Raper-Mason pattern appeared. The results show that, in the presence of PEG, tyrosinase-catalyzed oligomerization of DOPA exhibits kinetics slower than those observed in the absence of the polymer. However, melanogenesis still takes place in the presence of PEG, as indicated by the formation of black pigments and by the detection of DHI and DHICA oligomers, considered to be the first intermediates in melanin formation.