The "Mellanby effect" in alcoholised e-scooter drivers.

PURPOSE: Several studies tried to discuss and clarify the so-called Mellanby effect: Similar blood alcohol concentrations (BACs) supposedly lead to more signs of impairment in the phase of alcohol resorption than elimination. To assess this effect for alcoholised e-scooter driving, results of a real-driving fitness study were subanalysed. METHODS: Sixteen subjects (9 females; 7 males) who completed runs at comparable BACs in the phases of alcohol resorption and elimination were chosen to assess a possible "Mellanby effect". The data of the subjects was taken from a prior e-scooter study by Zube et al., which included 63 subjects in total. RESULTS: In the phase of alcohol resorption, the relative driving performance was approx. 92% of the phase of elimination (p value 0.21). Statistically significant more demerits were allocated to the obstacle "narrowing track" in the phase of resorption than elimination. Subjects also needed significantly more time to pass the obstacles "narrowing track", "driving in circles counterclockwise" and "thresholds" in the phase of resorption than elimination. DISCUSSION: The most relevant obstacle to discriminate between the two different states of alcoholisation was the narrowing track. Insofar, measurements of the standard deviation of the lateral position (SDLP) might also be a sensitive component for the detection of central nervous driving impairment during shorter trips with an e-scooter. Additionally, driving slower during the phase of alcohol resorption seems to be the attempt to compensate alcohol-related deficits. CONCLUSION: The results of the study suggest a slight Mellanby effect in e-scooter drivers.

Contribution of cyclooxygenase-1-dependent prostacyclin synthesis to bradykinin-induced dermal extravasation.

BACKGROUND: Non-allergic angioedema is a potentially life-threatening condition caused by accumulation of bradykinin and subsequent activation of bradykinin type 2 receptors (B2). Since COX activity plays a pivotal role in B2 signaling, the aim of this study was to determine which prostaglandins are the key mediators and which COX, COX-1 or COX-2, is predominantly involved. METHODS: We used Miles assays to assess the effects of inhibitors of COX, 5-lipoxygenase, epoxyeicosatrienoic acid generation, cytosolic phospholipase A2α and a variety of prostaglandin receptor antagonists on bradykinin-induced dermal extravasation in C57BL/6 and COX-1-deficient mice (COX-1-/-). In addition, the prostacyclin metabolite 6-keto-PGF1α was quantified by ELISA in subcutaneous tissue from C57BL/6 and human dermal microvascular endothelial cells. In the latter, 6-keto-PGF1α was also quantified and identified by LC-MS/MS. RESULTS: Unspecific COX inhibition by ibuprofen and diclofenac significantly reduced B2-mediated dermal extravasation in C57BL/6 but not COX-1-/-. Likewise, inhibition of cytosolic phospholipase A2α showed similar effects. Furthermore, extravasation in COX-1-/- was generally lower than in C57BL/6. Of the prostaglandin antagonists used, only the prostacyclin receptor antagonist RO1138452 showed a significant reduction of dermal extravasation. Moreover, 6-keto-PGF1α concentrations were increased after bradykinin treatment in subcutaneous tissue from C57BL/6 as well as in human dermal microvascular endothelial cells and this increase was abolished by diclofenac. CONCLUSION: Our findings suggest that COX-1-dependent prostacyclin production is critically involved in dermal extravasation after activation of B2 in small dermal blood vessels. Targeting prostacyclin production and/or signaling appears to be a suitable option for acute treatment of non-allergic angioedema.

E-scooter driving under the acute influence of alcohol-a real-driving fitness study.

PURPOSE: To assess the effects of alcohol on the ability to drive an e-scooter, driving tests reflecting real-life situations accompanied by medical examinations focusing on balance were conducted at different blood alcohol concentrations (BACs). METHODS: Fifty-seven subjects who consumed alcohol (28 female, 29 male) and 6 consistently sober subjects (3 female, 3 male) participated in the study. Alcohol was administered on a fixed schedule, and the individual drinking quantity was individually calculated in advance using the Widmark formula. Repeated runs through a fixed course were performed. Following each ride, a blood sample was taken for BAC determination, and medical tests were performed. RESULTS: Even at low BACs (0.21-0.60 g/kg), subjects showed a significant decrease in driving performance, to approximately 60% of the initial level. Differences in driving performance at different BAC ranges were observed for different obstacles, especially for the narrowing track, gate passage, slalom, and driving in circles obstacles. Furthermore, worse Romberg and Unterberger test results were correlated with worse driving performance. It cannot be assumed that learning effects during the study had a relevant effect, as shown in the comparison of the driving performance of the alcohol-consuming group with that of the control group. Sex-specific differences were not found. DISCUSSION: Significant deteriorations in driving performance at BACs below 1.10 g/kg confirmed alcohol-related risk potential when using e-scooters. At this time, these findings may lead to the assumption of "relative driving impairment" in Germany. The Romberg and Unterberger tests could be considered a complementary investigation method for the assessment of e-scooter driving impairment. CONCLUSION: Even at rather low BACs between 0.21 and 0.40 g/kg, there was a significant deterioration in driving performance under the influence of alcohol compared to sober, which highlights the negative effects of alcohol on e-scooter driving.

Pupillary effects in habitual cannabis consumers quantified with pupillography.

Driving under the influence of alcohol (DUIA) and drugs (DUID) is considered an elevated risk for traffic safety. When assessing a driver's fitness to drive, standardized and objective measurement methods are still required, in order to clarify the question whether an individual is under the influence of substances acting on the central nervous system (CNS). We exposed healthy test subjects (n=41) as well as persons who were under the influence of cannabis after repeated inhalation to multiple light stimuli using infrared technology and measured the pupillary light reflex (PLR). Toxicological tests of blood samples taken from every subject followed. The aims of this study were to assess the differences in pupillography response between cannabis consumers after a washout period and no cannabis consumers as well as the dose related effects on pupillography parameters of cannabis in cannabis consumers. All four pupillary parameters changed according to a weakened pupil function after acute administration of cannabis in all test subjects. Furthermore, it could be observed that habitual cannabis consumers showed an altered pupillary function just before the first dose was taken, suggesting that the long-term effects and addiction also have to be taken into account, when effects of the CNS are discussed. The results of the present study show that almost all pupil parameters could be reliable indicators for the detection of subjects under the acute effect of cannabis.

Ophthalmologic examinations under the acute influence of alcohol.

Alcohol is the most widely used recreational drug in Western countries. It affects the psychophysical performance in different ways, e.g. by reducing cognitive functions, causing coordination disturbances or impairing vision. Visual impairments both concern oculomotor and visual sensory functions, such as decreased mesopic vision, decreased field of vision and an increase of saccadic eye movements. During cycling trials with alcoholised test persons, repeated measurements of (1.) the time needed to read a 50-word text, (2.) the time to perform a swing test by tenfold touching the moving fingertip of the examiner, and (3.) the amplitude of fusion were carried out. The results of these tests were further evaluated to test the hypothesis that impaired vision is significantly correlated to reduced cycling performances of alcoholised persons. In a second step, it was examined which test is most useful to identify alcohol intoxicated cyclists. The ophthalmologic examination results of the groups of best and worst cycling-performing test persons at blood alcohol levels between 0.10% and 0.15% were set into relation to the documented allocated demerits. Additionally, the individual results of these persons were compared to the state of soberness. The time needed to read a 50-word text significantly correlated with the cycling performance. As this is an easy and objective test, it might contribute to a synoptic evaluation of the psychophysical performance of a drunken cyclist.

On the impact of cannabis consumption on traffic safety: a driving simulator study with habitual cannabis consumers.

To contribute to the ongoing discussion about threshold limits of Δ9-tetrahydrocannabinol (THC) in road traffic, a driving simulator study with 15 habitually cannabis consuming test persons was conducted. Probands were tested on different routes after consumption of a maximum of three cannabis joints, each containing 300 µg THC/kg body weight (sober testing as well as testing directly, 3 and 6 h after cannabis consumption). Accompanying the drives, medical examinations including a blood sampling were performed. Driving faults and distinctive features in the medical examinations were allocated certain penalty points, which were then summed up and evaluated using the ANOVA model. The results showed that very high CIF values > 30 as well as serum THC concentrations > 15 ng/ml significantly increased the number of penalty points, but no direct correlation to the THC concentrations in serum and/or CIF values was detected. Instead, the point in time after cannabis consumption seems to play an important role concerning driving safety: significantly more driving faults were committed directly after consumption. Three hours after consumption, no significant increase of driving faults was seen. Six hours after consumption (during the so-called subacute phase), an increase of driving faults could be noted although not significant. Considering the limitation of our study (e.g. small test group, no placebo test persons, long lasting test situation with possible tiredness), further studies focusing on the time dependant impact of cannabis consumption on road traffic are required.

Analysis of cocaine adulterants in human brain in cases of drug-related death.

For different reasons, street cocaine is often diluted with pharmacologically active substances, the so-called adulterants such as levamisole or hydroxyzine. A controversial debate exists currently on the uptake of adulterants from cocaine preparations and drug-related death. Previous research convincingly argues that serious adverse side effects that affect the central nervous and cardiovascular systems can be a consequence of adulterated cocaine. AIMS: Having identified the presence of adulterants in lung tissue and blood, the concentrations of these substances in brain, an important target location, was of interest. This provides an opportunity to assess their role in cases of drug-related deaths. MATERIALS AND METHODS: We developed and validated a method for the analysis of cocaine, two cocaine metabolites and six adulterants, which can typically be found in cocaine preparations, and one adulterant metabolite in brain tissue by gas chromatography-mass spectrometry (GC-MS)1. Ten brain samples which were tested positive for cocaine were analyzed. The homogenized brain tissue was embedded into drying paper for protein precipitation. During a subsequent solid-phase extraction (SPE), the eluate and one of the wash fractions were collected. After derivatization with N-Methyl-N-(trimethylsilyl)trifluoroacetamide (MSTFA) in pyridine and isooctane, the extracts were analyzed by GC-MS. RESULTS AND DISCUSSION: The method was fully validated for cocaine (COC), benzoylecgonine (BZE), ecgonine methyl ester (EME), diltiazem (DIL), hydroxyzine (HYD), and levamisole (LEV) and partly validated for cetirizine (CET), lidocaine (LID), phenacetin (PHE), and procaine (PRO) in brain material. By analyzing post-mortem brain tissue of ten cocaine users, LEV, LID, and HYD as well as PHE were identified in contrast to DIL, PRO, and the HYD metabolite CET. HYD and LEV were found in moderate to high concentrations in some cases. Therefore, it cannot be excluded that they have caused adverse side effects. CONCLUSION: Because adulterants can potentially affect the central nervous and cardiac systems, it is likely that they enhance COC toxicity.

Indole-3-carbinol, a plant nutrient and AhR-Ligand precursor, supports oral tolerance against OVA and improves peanut allergy symptoms in mice.

In general, dietary antigens are tolerated by the gut associated immune system. Impairment of this so-called oral tolerance is a serious health risk. We have previously shown that activation of the ligand-dependent transcription factor aryl hydrocarbon receptor (AhR) by the environmental pollutant 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) affects both oral tolerance and food allergy. In this study, we determine whether a common plant-derived, dietary AhR-ligand modulates oral tolerance as well. We therefore fed mice with indole-3-carbinole (I3C), an AhR ligand that is abundant in cruciferous plants. We show that several I3C metabolites were detectable in the serum after feeding, including the high-affinity ligand 3,3´-diindolylmethane (DIM). I3C feeding robustly induced the AhR-target gene CYP4501A1 in the intestine; I3C feeding also induced the aldh1 gene, whose product catalyzes the formation of retinoic acid (RA), an inducer of regulatory T cells. We then measured parameters indicating oral tolerance and severity of peanut-induced food allergy. In contrast to the tolerance-breaking effect of TCDD, feeding mice with chow containing 2 g/kg I3C lowered the serum anti-ovalbumin IgG1 response in an experimental oral tolerance protocol. Moreover, I3C feeding attenuated symptoms of peanut allergy. In conclusion, the dietary compound I3C can positively influence a vital immune function of the gut.

Fingolimod promotes peripheral nerve regeneration via modulation of lysophospholipid signaling.

BACKGROUND: The lysophospholipids sphingosine-1-phosphate (S1P) and lysophosphatidic acid (LPA) are pleiotropic signaling molecules with a broad range of physiological functions. Targeting the S1P1 receptor on lymphocytes with the immunomodulatory drug fingolimod has proven effective in the treatment of multiple sclerosis. An emerging body of experimental evidence points to additional direct effects on cells of the central and peripheral nervous system. Furthermore, fingolimod has been reported to reduce LPA synthesis via inhibition of the lysophospholipase autotaxin. Here we investigated whether modulation of particular signaling aspects of S1P as well as LPA by fingolimod might propagate peripheral nerve regeneration in vivo and independent of its anti-inflammatory potency. METHODS: Sciatic nerve crush was performed in wildtype C57BL/6, in immunodeficient Rag1 (-/-) and Foxn1 (-/-) mice. Analyses were based on walking track analysis and electrophysiology, histology, and cAMP formation. Quantification of different LPA species was performed by liquid chromatography coupled to tandem mass spectrometry. Furthermore, functional consequences of autotaxin inhibition by the specific inhibitor PF-8380 and the impact of fingolimod on early cytokine release in the injured sciatic nerve were investigated. RESULTS: Clinical and electrophysiological measures indicated an improvement of nerve regeneration under fingolimod treatment that is partly independent of its anti-inflammatory properties. Fingolimod treatment correlated with a significant elevation of axonal cAMP, a crucial factor for axonal outgrowth. Additionally, fingolimod significantly reduced LPA levels in the injured nerve. PF-8380 treatment correlated with improved myelin thickness. Sciatic nerve cytokine levels were not found to be significantly altered by fingolimod treatment. CONCLUSIONS: Our findings provide in vivo evidence for direct effects of fingolimod on cells of the peripheral nervous system that may propagate nerve regeneration via a dual mode of action, differentially affecting axonal outgrowth and myelination by modulating relevant aspects of S1P and LPA signaling.

A sensitive method for the determination of hordenine in human serum by ESI⁺ UPLC-MS/MS for forensic toxicological applications.

We present the determination of the alkaloid hordenine and its forensic relevance as a qualitative and quantitative marker for beer consumption. A simple, rapid and sensitive ultra-performance liquid chromatography (UPLC)-tandem mass spectrometry (MS/MS) method for the determination of hordenine in human serum samples was developed and validated. The application was tested with serum samples after enzymatic cleavage. After addition of the synthesized internal standard hordenine-D 4, a liquid-liquid extraction with dichloromethane and diethyl ether was performed. Chromatographic separation was conducted with a Waters Acquity® UPLC system with gradient elution on an Agilent Eclipse XDB-C18 column (4.6 mm × 150 mm, 5-µm particle size). For quantification, a Waters Acquity® TQ detector (version SNC 627) with a positive electrospray ionization probe and multiple reaction monitoring mode was used. A flow rate of 0.4 ml/min was applied. The retention time for both the analyte and the internal standard was 3.67 min. Linearity was demonstrated from 0.2 to 16 ng/ml (R(2) > 0.999). The lower limit of quantification was 0.3 ng/ml in serum. Matrix effects and extraction recoveries for low and high concentrations were within acceptable limits of 75-125% and 50%, respectively. To the best of our knowledge there is no corresponding method for the determination of hordenine by UPLC-MS/MS in serum. By our drinking studies we demonstrate that beer consumption leads to detectable hordenine concentrations in serum and observed a linear elimination of total hordenine correlating to blood alcohol concentration, which shows that hordenine can be used as a reliable qualitative and quantitative marker for beer consumption. The validated method was successfully applied to serum from actual forensic cases.

Ornithine transcarbamylase deficiency of a male newborn with fatal outcome.

Ornithine transcarbamylase deficiency (OTCD) is the most common malfunction of ureagenesis. The case of a male newborn who died at the age of 2 days for clinically unclear reasons is presented. The post-mortem routine and esoteric testing methods that finally led to the diagnosis of a fatal case of OTCD are outlined here.

The effect of cannabis on regular cannabis consumers' ability to ride a bicycle.

To assess the effects of cannabis on the ability required to ride a bicycle, repetitive practical cycling tests and medical examinations were carried out before and after inhalative consumption of cannabis. A maximum of three joints with body weight-adapted THC content (300 µg THC per kg body weight) could be consumed by each test subject. Fourteen regular cannabis-consuming test subjects were studied (12 males, 2 females). In summary, only a few driving faults were observed even under the influence of very high THC concentrations. A defined THC concentration that leads to an inability to ride a bicycle cannot be presented. The test subjects showed only slight distinctive features that can be documented using a medical test routinely run for persons under suspicion of driving under the influence of alcohol or drugs.

Comparison of venous blood alcohol concentrations and breath alcohol concentrations measured with Draeger Alcotest 9510 DE Evidential.

Most comparisons of blood alcohol concentrations (BAC) and breath alcohol concentrations (BrAC) are either derived from drinking trials with rigid drinking protocols or from investigative authorities' data with considerable time differences between the determination of BAC and BrAC. In general, only comparisons of relatively low BAC-BrAC pairs are available. Therefore, the relationship between BAC and BrAC was examined even for high BAC above 2g/kg. The results of a large-scale drinking test under realistic conditions with 78 test persons and short time intervals between BAC and BrAC measurements are presented. It was shown that the conversion factor Q varies greatly (between 1571:1 and 2394:1) and increases with increasing BAC. A constant conversion factor that is suitable for variable forensic purposes could not be presented.

Abuse of the over-the-counter antispasmodic butylscopolamine for the home synthesis of psychoactive scopolamine.

UNLABELLED: We report on two patients who ingested psychoactive scopolamine that was synthesized at home from butylscopolamine (Buscopan®), which is available as over-the-counter antispasmodic in nearly 100 countries worldwide. Patient 1 presented with severe central anticholinergic toxidrome, while patient 2 suffered from minor symptoms. An empty blister of Buscopan® was found in the patients' home, but initially was not suspected to be causative for the observed central anticholinergic symptoms, as Buscopan® is not able to pass the blood-brain barrier in its native form. Only later, the information by third parties and a Google search helped to identify homemade scopolamine derived from Buscopan® as the responsible agent in these two cases. Retrospectively, scopolamine could be detected in serum and urine of both patients, while it was absent in one control after ingestion of native Buscopan®. CONCLUSION: Over-the-counter drugs can be used to synthesize psychoactives with means that are available in every household. Such knowledge can spread via social media and internet discussion boards long before appearing in medical literature. While typical clinical presentation often enables clinicians to adequately identify and treat specific toxidromes, these sources of information need to be increasingly taken into account by medical professionals for identification of its causative agent. This potential of Buscopan® might gain importance as an easily accessible source of psychoactive scopolamine. WHAT IS KNOWN: • Substances with central anticholinergic effects are known for their hallucinogenic potential and may be used as psychoactives. What is New: • The over-the-counter antispasmodic butylscopolamine (Buscopan®) can be abused to synthesize anticholinergic, psychoactive scopolamine at home with means that are available in every household.

The effect of alcohol hangover on the ability to ride a bicycle.

To investigate the effects of alcohol on the ability to ride a bicycle, practical cycling tests were carried out at different blood alcohol concentrations (BAC). For this purpose, various alcoholic beverages could be consumed from around 2 p.m. until 11 p.m. Afterwards, the test persons spent the night on the trial site and were provided with dormitory sleeping accommodation. On the following morning, beginning at around 8 a.m., a final cycling test was performed. The performances of those test persons who had returned to state of soberness and of those with residual blood alcohol levels were compared to the performances on the day before. The practical ability to ride a bicycle was significantly reduced in the postalcoholic state compared to the rides of the day before. The relative cycling performance in the postalcoholic state was comparable to the rides under the influence of BAC of around 0.30 g/kg. There were no remarkable differences between the groups with and without residual blood alcohol levels regarding the rides on the next morning. Therefore, it can be assumed that the direct influence of residual blood alcohol levels plays a minor role for the ability to ride a bicycle in the postalcoholic state. Instead, the side effects of the high amounts of alcohol that were consumed the night before are crucial.

Differences between male and female cyclists' performances under the acute influence of alcohol.

To examine the effects of alcohol regarding the fitness required to ride a bicycle, practical cycling tests, accompanied by medical examinations, were carried out at different blood alcohol concentrations. Seventy-eight persons were included in the trials (41 males, 37 females). Eighty-three evaluable trials were obtained. Men committed less coordinative driving faults with comparable blood alcohol concentrations. Single highly alcoholized men were able to safely ride their bicycle; however, each of the female test persons had at least one severe driving fault at blood alcohol levels above 1.43 g/kg. Women tended to exhibit signs of alcoholization in the medical examination reports earlier than men.

Drug-related death: adulterants from cocaine preparations in lung tissue and blood.

The abuse of drugs such as street cocaine is known to cause a variety of toxic effects, some of which involve the lungs and often induce lethal complications. While the toxicity of cocaine itself is reviewed well, the influence of toxic effects of its adulterants on the human body is not thoroughly studied. Therefore, we examined heart blood, femoral vein blood and lung tissue from 11 cases for typically used adulterants in cocaine preparations and check whether if the concentrations in the lung tissue are higher than in the blood. The adulterants were isolated using solid-phase (SPE) and liquid-liquid extraction (LLE) and quantified via high-pressure-liquid-chromatography-time-of-flight-mass spectrometry (LC/TOF-MS). Five adulterants, i.e., phenacetin, lidocaine, diltiazem, levamisole and hydroxyzine, were detected. We found out that the concentration of these substances was often higher in the lung than in the analogous analysed body fluids. It should therefore be considered whether - for the determination in the cause of death - the lung should be examined in addition to heart blood, urine or brain tissue.

Regarding the fitness to ride a bicycle under the acute influence of alcohol.

To determine the threshold for the absolute inability to ride a bicycle, practical cycling tests and medical examinations at different blood alcohol concentrations were performed. Special attention was given to additional medical examinations, reaction tests and alcohol consumption under real-life conditions. Seventy-eight test subjects were included in the trials (37 females, 41 males). Five test subjects participated twice; thus, there were a total of 83 evaluable trials. Alcohol-related deficits were already identifiable at very low BACs. A significant increase in gross motoric disturbances compared to the soberness state did not regularly occur until a BAC of at least 0.8 g/kg was reached. At the BAC of 1.4 g/kg and above, no test subjects were able to achieve or surpass their sober driving results. Isolated highly alcoholised test subjects rode the bike in a manner that was not conspicuously different than the other sober test persons. Contrary to the assumptions of current German legal practise, it cannot be stated that all people are 'absolutely impaired' to the point of being incapable of riding bicycle at BACs of at least 1.6 g/kg.

Comprehensive automation of the solid phase extraction gas chromatographic mass spectrometric analysis (SPE-GC/MS) of opioids, cocaine, and metabolites from serum and other matrices.

The analysis of opioids, cocaine, and metabolites from blood serum is a routine task in forensic laboratories. Commonly, the employed methods include many manual or partly automated steps like protein precipitation, dilution, solid phase extraction, evaporation, and derivatization preceding a gas chromatography (GC)/mass spectrometry (MS) or liquid chromatography (LC)/MS analysis. In this study, a comprehensively automated method was developed from a validated, partly automated routine method. This was possible by replicating method parameters on the automated system. Only marginal optimization of parameters was necessary. The automation relying on an x-y-z robot after manual protein precipitation includes the solid phase extraction, evaporation of the eluate, derivatization (silylation with N-methyl-N-trimethylsilyltrifluoroacetamide, MSTFA), and injection into a GC/MS. A quantitative analysis of almost 170 authentic serum samples and more than 50 authentic samples of other matrices like urine, different tissues, and heart blood on cocaine, benzoylecgonine, methadone, morphine, codeine, 6-monoacetylmorphine, dihydrocodeine, and 7-aminoflunitrazepam was conducted with both methods proving that the analytical results are equivalent even near the limits of quantification (low ng/ml range). To our best knowledge, this application is the first one reported in the literature employing this sample preparation system.