Hair analysis in order to evaluate drug abuse in driver's license regranting procedures.

In Italy, driving under the influence of drugs determines the suspension of the offender's driver's license. To regain the license the person must be drug free during an observation period. People whose license has been revoked or suspended can obtain, or re-obtain their driver's license subject to the judgment of a medical commission. The exclusion of illicit drug use is determined by means of toxicological analysis, mainly on urine or hair matrices. We reported the results of several years of experience of the forensic toxicology laboratory of the University of Macerata in the use of hair analysis for the assessment of past exposure to drugs in people suspected of driving under the influence of drugs. From 2004 to 2013, 8612 hair samples, were analyzed for opiates, cocaine and delta-9-tetrahydrocannabinol (Δ(9)-THC) using gas chromatography/mass spectrometry (GC/MS) method. We used a cutoff (SoHT or national guidelines) to determine the positive data, regardless of the hair sample concentrations. 1213 samples resulted positive, 71.7% were positive for cocaine and metabolites, 19.8% for morphine and metabolites, 8.5% for Δ(9)-THC. We also studied the timeframe of the abuse, as well as gender and age distribution of positive subjects. Moreover, we analyzed the possible deterrent effect of the hair analysis on driving under the influence of psychoactive substances.

Preliminary gas chromatography with mass spectrometry determination of 3,5-dimethoxyphenol in biological specimens as evidence of taxus poisoning.

Taxus baccata is a widely distributed yew often associated with cases of fatal intoxication, which is related to the high amounts of cardiotoxic alkaloids, taxine A and taxine B, contained in its leaves. In this paper, a case of Taxus fatal poisoning, hypothesized by the forensic autopsy, has been confirmed by the application of both gas chromatography-mass spectrometry (GC-MS) and liquid chromatography-tandem mass spectrometry (LC-MS-MS) techniques. A GC-MS method was used for the determination of the derivatized 3,5-dimethoxyphenol, a cyanogenic aglylactone considered as a marker of Taxus poisoning, being present in all species of Taxus. The detection by LC-MS-MS of taxine B and isotaxine B in the biological specimens confirmed the absorption of cardiotoxic alkaloids and allowed to established the cause of death as the ingestion of Taxus baccata leaves.

A case of suicide by ingestion of sodium nitroprusside.

The study reports a case of suicide by ingestion of sodium nitroprusside which resulted in acute cyanide poisoning. Analyses carried out on body fluid yielded the quantitation of total (5.00 mg/L) and free (3.30 mg/L) cyanide in blood and of methemoglobin (blood = 10.5%). At the scene, some solid reddish-brown material was found in a glass, which on toxicological analysis was found to contain sodium nitroprusside; about 9 g of the same substance was identified in stomach contents. The detection and quantification of cyanide and methemoglobin in biological samples from the case indicated that the lethal effect was due to both metabolic products (cyanide and methemoglobin).

Effects of a methanolic extract and a hyperforin-enriched CO2 extract of Hypericum perforatum on alcohol intake in rats.

Hypericum perforatum extracts (HPE) inhibit ethanol intake in rats. Hypericin and hyperforin have been proposed as major active principles of HPE. The present study compared the effect on ethanol intake in alcohol-preferring rats of two Hypericum perforatum extracts: a methanolic extract containing 0.3% hypericin and 3.8% hyperforin (HPE1) and a CO2 extract (HPE2) with 24.33% hyperforin and very low hypericin content. Freely feeding and drinking rats were offered 10% ethanol 2 h/day and HPE were given intragastrically 1 h before access to ethanol. Both extracts dose-dependently reduced ethanol intake, HPE2 being about eight times more potent than HPE1. Food and water intakes were not affected by doses that reduced ethanol intake. HPE2, unlike HPE1, reduced blood-alcohol levels (BAL) at doses of > or = 31.2 mg/kg, whereas the dose of 15.6 mg/kg, which reduced ethanol intake, did not significantly modify BAL; blood-acetaldehyde levels were never increased. As previously observed for HPE1, intracerebroventricular pretreatment with 5,7-dihydroxytryptamine (150 microg/rat) did not affect attenuation of ethanol intake induced by HPE2, but reduced its effect in the forced swimming test (FST). Intraperitoneal pretreatment with the sigma-1 receptor antagonist NE-100 (0.25 mg/kg) did not affect inhibition of ethanol intake induced by HPE1 (250 mg/kg) or HPE2 (125 mg/kg), but abolished the effect of both extracts in the FST. In conclusion, the present results indicate that HPE2 inhibits ethanol intake more potently than HPE1; the higher potency of HPE2 parallels the hyperforin content, suggesting that hyperforin may have an important role in reducing ethanol intake. Moreover, different neurochemical mechanisms are apparently responsible for the reduction of ethanol intake and for the antidepressant-like effect of HPE.

Detection and quantitation of morphine in fixed tissues and formalin solutions.

The study reports the results of the detection and quantitation of morphine in tissues fixed in formalin and in the formalin solutions in which the same tissues were fixed. Toxicological analyses were performed on formalin-fixed liver and kidney samples from five cases of opiate poisoning and on the formalin solutions (10% buffered pH 7) in which the samples were preserved. Analyses carried out at the time of autopsy on body fluids and tissues allowed identification of opiate as the cause of death and its quantitation. Tissue samples were preserved in formalin solutions for 12 weeks before analysis. The mean levels of recovery of morphine in fixed tissues were 36.29% in liver, 29.41% in kidney, 74.93% in formalin from liver, and 42.17% in formalin from kidney. Results indicated that this particular toxic substance shows good stability even in biological specimens subjected to chemical fixation.

[Poisons and men: historical, legal and toxicological aspects]. / Il veleno e l'uomo: aspetti storici, giuridici e tossicologici.

Humanity has always practised the poisoning. At first, magic and mistery characterized this ritual. Then, thanks to a more careful legislation and to empirical research it became subject of toxicology.

Conditioned taste aversion induced by ethanol in alcohol-preferring rats: influence of the method of ethanol administration.

A recent study of our group has shown that ethanol evokes conditioned place preference (CPP) in Marchigian Sardinian alcohol-preferring (msP) rats following intragastric (IG) administration by means of an indwelling IG catheter, but not following administration by gavage or by intraperitoneal (IP) injection. The present study evaluated in ethanol-naive msP rats the influence of the method of administration (IG injection by indwelling catheter vs. IP injection) on ethanol-induced conditioned taste aversion (CTA). The dose of 0.35 g/kg of ethanol did not evoke aversion either by IG or by IP administration. Following IG injection, 0.7 g/kg of ethanol, the amount that msP rats voluntarily ingest in a short (2-5 min) drinking episode, did not evoke CTA, and 1.5 g/kg induced a modest CTA. On the other hand, IP injection of 0.7 g/kg of ethanol evoked CTA, and 1.5 g/kg induced a very pronounced CTA. These findings show that the aversive properties of ethanol in msP rats are influenced by the method of administration, and suggest that the IG injection by catheter may reveal more faithfully than the IP injection the motivational properties of amounts of ethanol that alcohol-preferring rats voluntarily ingest.

Antidepressant-like effect of ethanol revealed in the forced swimming test in Sardinian alcohol-preferring rats.

RATIONALE: A large body of evidence indicates high comorbidity between depression and alcohol abuse. The self-medication hypothesis proposes that depressed subjects may abuse ethanol because it reduces the symptoms of depression. The present study evaluated whether ethanol may exert an antidepressant-like action in genetically selected alcohol-preferring rats, either Sardinian alcohol-preferring (sP) or Marchigian Sardinian alcohol-preferring (msP) rats, and for comparison in Sardinian alcohol-non-preferring (sNP) rats. METHODS: The forced swimming test (FST) was used to evaluate the antidepressant-like action of ethanol; in this test the effect of ethanol ingestion on the immobility time was determined. RESULTS: Ethanol-naive sP rats exhibited a longer period of immobility in comparison to sNP rats. Both in ethanol-naive sP and msP rats, voluntary ethanol drinking reduced the immobility time. A similar effect was obtained when repeated (five or nine) intragastric administrations of 0.7 g/kg ethanol were given during the 24 h prior to the test in msP and in sP, but not in sNP rats. Desipramine, like ethanol, sharply reduced immobility at doses of 5 or 20 mg/kg, given 3 times in the 24 h before the test in msP rats. The reduced immobility induced by ethanol in msP rats was apparently not the consequence of a general motor activation, because 9 IG administrations of ethanol, 0.7 g/kg, failed to alter locomotor activity in the open field test. Moreover, blood alcohol levels and rectal temperature of msP, sP and sNP after IG ethanol administration were not statistically different. CONCLUSIONS: The present results provide evidence for an antidepressant-like action of ethanol in sP and msP rats and suggest that this action may contribute to sustain their high ethanol drinking.

Analytical detection and quantitation of strychnine in chemically fixed organ tissues.

This study reports the results of the detection and quantitation of strychnine in formalin-fixed tissues and in the formalin solutions in which the tissues were fixed. The toxicological analyses were performed on formalin-fixed liver and kidney samples and formalin solutions (10% buffered pH 7) in which the same samples from a case of acute strychnine poisoning were preserved. The analyses carried out at the time of autopsy on body fluid and tissues (bile, 2.40 mg/L; stomach contents, 14.2 mg; liver, 6.68 mg/kg; kidney, 2.68 mg/kg) allowed the identification of this substance as cause of death. The tissue samples were preserved in formalin solutions for 8 weeks. The analyses performed on formalin-fixed tissues (liver and kidney) and on formalin solutions, in which the same tissues were preserved, permitted the detection and quantitation of strychnine (liver, 1.59 mg/kg; formalin from the liver, 1.80 mg/L; kidney, 0.98 mg/kg; formalin from the kidney, 1.11 mg/L). The results indicate that this particular toxic substance also shows good stability in biological specimens subjected to chemical fixation.

Ethanol induces conditioned place preference in genetically selected alcohol-preferring rats.

The study of the biological mechanisms of ethanol reward has greatly suffered from problems to obtain ethanol-induced conditioned place preference (CPP) in rats. In the present study, CPP was obtained in genetically selected Marchigian Sardinian alcohol-preferring (msP) rats, derived from Sardinian alcohol-preferring rats, following intragastric (IG) ethanol administration by means of a permanent IG catheter, but not after intraperitoneal (IP) injection or IG gavage. Rats with permanent IG catheter, received IG administration of 0.35, 0.7, 1.5 or 2.8 g/kg ethanol, as a 10% v/v solution. In ethanol-experienced rats 0.7 or 1.5, but not 0.35 or 2.8 g/kg ethanol significantly increased in comparison to controls the time spent in the ethanol-associated previously non-preferred compartment, which became preferred in the post-conditioning test. In ethanol-naive rats, only 0.7 g/kg ethanol significantly increased the time spent in the ethanol-associated compartment. On the other hand, no effect was observed in alcohol-experienced rats following IG gavage, or IP injection of 0.35, 0.7 or 1.5 g/kg ethanol. The present results provide evidence that ethanol possesses postingestive rewarding properties in msP rats, and that it can reliably induce CPP in them, provided that an appropriate method of administration is adopted.

Mechanism of action for reduction of ethanol intake in rats by the tachykinin NK-3 receptor agonist aminosenktide.

Intracerebroventricular (i.c.v.) injection of tachykinin (TK) NK-3 receptor agonists inhibits alcohol intake in genetically selected alcohol-preferring rats. The present study investigated the mechanism of action by which the selective TK NK-3 receptor agonist aminosenktide (NH2-SENK) attenuates ethanol intake in Marchigian Sardinian alcohol-preferring (msP) rats. The effect of NH2-SENK was studied by i.c.v. injection in the conditioned taste aversion (CTA) and in the conditioned place preference (CPP) paradigms; moreover, the effect of NH2-SENK on blood alcohol levels (BAL) following intragastric ethanol administration was investigated. The i.c.v. dose of 125 ng/rat of NH2-SENK, that markedly reduces ethanol intake, did not modify BAL, nor did it increase the CTA induced by intraperitoneal injection of ethanol, 1 g/kg body weight. These findings suggest that the effect of NH2-SENK on alcohol consumption is not related to modification of the pharmacokinetics of ethanol, nor to increase of the aversive properties of ethanol. On the other hand, the same i.c.v. dose of NH2-SENK evoked a pronounced and statistically significant CPP. This finding indicates that the TK NK-3 receptor agonist NH2-SENK possesses rewarding properties in msP rats and suggests that its inhibitory effect on ethanol consumption may be due to substitution of the rewarding properties of ethanol, thus making its consumption redundant.

Possible mechanism of action for the attenuation of ethanol intake induced by ritanserin in rats.

The 5-HT2 receptor antagonist, ritanserin, reduces alcohol intake in rats and the nucleus accumbens (NAC) has been proposed as a site of action for the drug. Recent microdialysis studies have shown that acute subcutaneous (SC) administration of ritanserin increases extracellular 5-HT levels in the NAC. The present study evaluated, in genetically heterogeneous rats with developed preference for 3% ethanol, whether the attenuation of ethanol intake induced by ritanserin might be related to its effect on the synaptic availability of 5-HT in the NAC. Damaging 5-HTergic neurons by intracerebroventricular infusion of 5,7-dihydroxytryptamine (5,7-DHT) abolished the effect of ritanserin on ethanol consumption. Injections of the 5-HT3 receptor antagonist MDL 72222 into the NAC significantly reduced the inhibitory effect of SC injection of ritanserin, 1 mg/kg, and completely abolished the effect of ritanserin, 0.1 mg/kg. Subcutaneous injections of MDL 72222, 0.3 mg/kg 3 times/day, suppressed the effect of SC ritanserin, 0.1 mg/kg. The present findings, together with those of previous experiments showing that the tryptophan hydroxylase inhibitor p-chlorophenylalanine abolishes the effect of ritanserin, support the hypothesis that its effect on ethanol intake may be due to increased synaptic availability of 5-HT into the NAC.