The Incidence of Dirofilaria immitis in Shelter Dogs and Mosquitoes in Austria.

To estimate the incidence of Dirofilaria immitis in Austrian shelter dogs and mosquitoes trapped in their proximity, 115 shelter dogs from fourteen animal shelters located in five different Austrian states were examined. Blood samples were screened for D. immitis using ELISA antigen-testing, PCR and microscopical examination for microfilariae. In total, 91% of the dogs originated from countries endemic for dirofilariosis. Eleven dogs (9.6%), all originating from Hungary, tested positive for D. immitis. None of the dogs examined showed microfilaremia. Eight dogs showed no or only mild clinical signs (e.g., infrequent coughing), and three dogs showed frequent coughing, dyspnea, exercise intolerance, blunt fur or weight loss. In total, 205 Mosquitoes of ten different species were caught at five different shelter sites in four different Austrian states, using CO2-baited mosquito traps set once a month (June-September 2019) for 24 h. All 205 mosquitoes tested negative for Dirofilaria spp. via PCR. The risk of endemisation of D. immitis in Austria (and other non-endemic countries in a similar situation) is very serious and its zoonotic potential should be communicated more strongly. To monitor a possible transmission of microfilariae from untreated or even untested positive dogs, e.g., in animal shelters, to mosquitoes in the near surroundings, frequent screening for Dirofilaria in mosquitoes should be used more intensively. Current knowledge on D. immitis should be integrated into daily veterinary practice and dog owners should be proactively educated, especially before traveling to endemic areas or adopting dogs from endemic countries. Animal shelters and animal welfare organizations should be provided with appropriate education and veterinary guidance regarding the testing and treatment of dogs imported from high-risk areas.

Scheduling of new psychoactive substance the Swiss way: A review and critical analysis.

Since the introduction of the European Early Warning System in 2005, >700 new psychoactive substances (NPS) have been listed. This review article presents for the first time the Swiss narcotic law in perspective of scheduling of NPS, and compares it to the regulations of the German speaking neighbours Austria and Germany. The Swiss way is a fast and effective way for scheduling NPS, with the purpose to restrict drug trafficking and for controlling the NPS drug market: the legal basis for scheduling substances of abuse is the "Law about narcotics and psychotropic substances" (BetmG, SR 812.121), which includes the "narcotic law directory (BetmVV-EDI, SR 812.121.11) suitable for listing all controlled substances. The BetmVV-EDI, SR 812.121.11 contains seven indices, with index e specifically designed for the fast scheduling of NPS. Newly appearing NPS can either be controlled under a structure analogues definition or by listing single substances. The list of single substances is updated at least once per year, and structure analogues definitions can be implemented, in order to keep track with new developments on the NPS market. The latest version from November 30th 2018 contains ten different structure analogue definitions and 207 single substances. Requirements to list NPS are their appearance on the NPS market, suspected psychotropic effects and their suggestions by Forensic professionals. As soon as substances are newly placed, on Schedule I of the 1961 Convention or Schedule II of the 1971 Convention by the Commission on Narcotic Drugs of the World Health Organization they can easily be transferred from index e to index a-d of the BetmVV-EDI, SR 812.121.11. The Austrian law uses a structure analogue and single substances approach (introduced in 2012, one update in 2016), whereas the German NPS law (established in 2016, no update yet) only lists two structure-analogue-definitions. All three legislations have defined which core structures, kinds and sites of substitutions are regulated.

Minimal Physiologically Based Pharmacokinetic Model of Intravenously and Orally Administered Delta-9-Tetrahydrocannabinol in Healthy Volunteers.

BACKGROUND AND OBJECTIVES: Lack of information on the pharmacokinetics of the active moiety of Cannabis or the metabolites of delta-9-tetrahydrocannabinol (THC) does not seem to be discouraging medical or recreational use. Cytochrome P450 (CYP) 2C9, the primary enzyme responsible for THC metabolism, has two single nucleotide polymorphisms-Arg144Cys (*2) and Ile359Leu (*3). In the Caucasian population, allelic frequency is between 0.08 and 0.14 for CYP2C9*2 and between 0.04 and 0.16 for CYP2C9*3. In vitro data suggest that metabolic capacity for the variants CYP2C9*2 and CYP2C9*3 is about one-third compared to wild-type CYP2C9. Previous work has suggested exposure to the terminal metabolite is genetically determined. We therefore sought to characterize the pharmacokinetics of THC and its major metabolites 11-hydroxy-delta-9-tetrahydrocannabinol (THC-OH) and 11-nor-9-carboxy-delta-9-tetrahydrocannabinol (THC-COOH) in healthy volunteers with known CYP2C9 status by non-compartmental analysis (NCA), compartmental modeling (CM) and minimal physiologically based pharmacokinetic (mPBPK) modeling. METHODS: Blood samples drawn for THC, THC-OH and THC-COOH after a single intravenous (IV) bolus of 0.1 mg/kg (0.32 µM/kg) THC were analyzed using a validated LC-MS/MS method. NCA generated initial estimates and CM and the mPBPK model were then fit to plasma concentration data using non-linear mixed-effects modeling. Blood samples from orally dosed (10, 25 and 50 mg) THC brownies were added to validate the model. RESULTS: THC can be described as a high hepatic extraction ratio drug with blood flow-dependent metabolism not restricted by protein binding. THC hepatic clearance is dependent on the CYP2C9 genetic variant in the population. High extraction drugs display route-dependent metabolism. When administered via the IV or inhalation routes, induction or inhibition of CYP2C9 should be non-contributory as the elimination of THC is dependent only on liver blood flow. THC-OH is also a high extraction ratio drug, but its hepatic clearance is significantly impacted by the hepatic diffusional barrier that impedes its access to hepatic CYP2C9. THC-COOH is glucuronidated and renally cleared; subjects homozygous for CYP2C9*3 have reduced exposure to this metabolite as a result of the polymorphism reducing THC production, the hepatic diffusional barrier impeding egress from the hepatocyte, and increased renal clearance. CONCLUSION: It has recently been reported that the terminal metabolite THC-COOH is active, implying the exposure difference in individuals homozygous for CYP2C9*3 may become therapeutically relevant. Defining the metabolism of THC in humans is important, as it is increasingly being used as a drug to treat various diseases and its recreational use is also rising. We have used NCA, CM, and mPBPK modeling of THC and its metabolites to partially disentangle the complexity of cannabis disposition in humans.

Simultaneous quantification of delta-9-THC, THC-acid A, CBN and CBD in seized drugs using HPLC-DAD.

An HPLC-DAD method for the quantitative analysis of Δ(9)-tetrahydrocannabinol (THC), Δ(9)-tetrahydrocannabinolic acid-A (THCA-A), cannabidiol (CBD), and cannabinol (CBN) in confiscated cannabis products has been developed, fully validated and applied to analyse seized cannabis products. For determination of the THC content of plant material, this method combines quantitation of THCA-A, which is the inactive precursor of THC, and free THC. Plant material was dried, homogenized and extracted with methanol by ultrasonication. Chromatographic separation was achieved with a Waters Alliance 2695 HPLC equipped with a Merck LiChrospher 60 RP-Select B (5µm) precolumn and a Merck LiChroCart 125-4 LiChrospher 60 RP-Select B (5µm) analytical column. Analytes were detected and quantified using a Waters 2996 photo diode array detector. This method has been accepted by the public authorities of Switzerland (Bundesamt für Gesundheit, Federal Office of Public Health), and has been used to analyse 9092 samples since 2000. Since no thermal decarboxylation of THCA-A occurs, the method is highly reproducible for different cannabis materials. Two calibration ranges are used, a lower one for THC, CBN and CBD, and a higher one for THCA-A, due to its dominant presence in fresh plant material. As provider of the Swiss proficiency test, the robustness of this method has been tested over several years, and homogeneity tests even in the low calibration range (1%) show high precision (RSD≤4.3%, except CBD) and accuracy (bias≤4.1%, except CBN).

Quantification of anandamide and 2-arachidonoylglycerol plasma levels to examine potential influences of tetrahydrocannabinol application on the endocannabinoid system in humans.

The effects of tetrahydrocannabinol (THC) and endogenous cannabinoids (endocannabinoids, ECs) are both mediated by activation of the cannabinoid receptors CB1 and CB2. Exogenous activation of these receptors by THC could therefore alter EC levels. We tested this hypothesis in healthy volunteers (n = 25) who received a large intravenous dose of THC (0.10 mg/kg). Effects on the EC system were quantified by serial measurements of plasma ECs after THC administration. Eleven blood samples were drawn during the first 5 h after THC administration and two more samples after 24 and 48 h. THC, its metabolites THC-OH (biologically active) and THC-COOH (non-active), and the ECs anandamide and 2-arachidonoylglycerol (2-AG) were quantified by liquid chromatography-mass spectrometry. EC-plasma levels showed a biphasic response after THC injection reaching maximal values at 30 min. Anandamide increased slightly from 0.58 ± 0.21 ng/ml at baseline to 0.64 ± 0.24 ng/ml (p < 0.05) and 2-AG from 7.60 ± 4.30 ng/ml to 9.50 ± 5.90 ng/ml (p < 0.05). After reaching maximal concentrations, EC plasma levels decreased markedly to a nadir of 300 min after THC administration (to 0.32 ± 0.15 ng/ml for anandamide and to 5.50 ± 3.01 ng/ml for 2-AG, p < 0.05). EC plasma concentrations returned to near baseline levels until 48 h after the experiment. THC (0.76 ± 0.16 ng/ml) and THC-OH (0.36 ± 0.17 ng/ml) were still measurable at 24 h and remained detectible until 48 h after THC administration. Although the underlying mechanism is not clear, high doses of intravenous THC appear to influence endogenous cannabinoid concentrations and presumably EC-signalling.

Trends and forecasts of hospital admissions for acute and chronic pancreatitis in the Netherlands.

BACKGROUND: The incidence and prevalence of acute and chronic pancreatitis have increased in Western countries. It is likely, the number of hospital admissions has increased correspondingly. AIMS: To analyze the trends in hospital admissions in the Netherlands for acute and chronic pancreatitis from 1992 to 2004 and to forecast the number of admissions up to 2010. METHODS: Analysis of hospital admissions for acute and chronic pancreatitis accumulated in a nationwide database. Curve fitting regression models were used to explore future trends. RESULTS: The number of acute pancreatitis admissions rose in 1992-2004 from 1,785 to 3,120 (74.8% increase). The overall 'annual number' of acute pancreatitis admissions increased from 11.8 to 19.2 per 100,000 person-years. The linear regression model predicted 3,205 [95% confidence intervals (CI), 3,111-3,299] and 3,537 (95% CI, 3,429-3,645) admissions for 2007 and 2010, respectively, a further increase of at least 9.9% in 2010 compared with 2004. In the 12-year time period, chronic pancreatitis admissions showed an increase of 75.4% (from 790 to 1,386). The overall 'annual number' of chronic pancreatitis admissions increased from 5.2 to 8.5 per 100,000 person-years. The cubic regression model predicted 1868 (95% CI, 1,619-2,117) and 3,173 (95% CI, 2,456-3,890) admissions for 2007 and 2010, respectively, an additional increase of 77.2% in 2010 compared with 2004. CONCLUSION: Hospital admissions for acute and chronic pancreatitis have increased substantially from 1992-2004. This trend will most likely continue for the near future and the burden and costs to the Dutch health care system will increase accordingly.

Application of headspace solid phase microextraction to qualitative and quantitative analysis of tobacco additives in cigarettes.

Cigarettes may contain up to 10% by weight additives which are intended to make them more attractive. A fast and rugged method for a cigarette-screening for additives with medium volatility was developed using automatic headspace solid phase microextraction (HS-SPME) with a 65 microm carbowax-divinylbenzene fiber and gas chromatography-mass spectrometry (GC-MS) with standard electron impact ionisation. In three runs, each cigarette sample was extracted in closed headspace vials using basic, acidic and neutral medium containing 0.5 g NaCl or Na2SO4. Furthermore, the method was optimized for quantitative determination of 17 frequently occurring additives. The practical applicability of the method was demonstrated for cigarettes from 32 brands.

Autoerotic accident by inhalation of propane-butane gas mixture.

We present a case of an accidental autoerotic death involving the inhalation of a propane-butane gas mixture, also known as LPG (liquefied petroleum gas). A 19-year-old male was found dead in supine position in his bed in a residential accommodation one day after he was last seen alive. On a personal computer at the end of the bed, a pornographic movie was still running. On his left shoulder, an empty rubber balloon and on the bedside 2 empty "Kisag-Gas" cartridges were found. Toxicologic investigations revealed an intoxication with propane and butane, together with a recent consumption of cannabis. This case report compares the toxicologic findings with other recently published cases, and the theories of the toxic effects are discussed.

Detection of cannabis use in drivers with the drugwipe device and by GC-MS after Intercept device collection.

Saliva or "oral fluid" has been presented as an alternative matrix in the establishment of drug exposure. The noninvasive collection of a saliva sample, which is relatively easy to perform and can be achieved under close supervision, is one of the most important benefits in a driving under the influence situation. Moreover, the presence of delta9-tetrahydrocannabinol (THC) in oral fluid is a better indication of recent use than when the drug is detected in urine, so there is a higher probability that the subject is experiencing pharmacological effects at the time of sampling. At 3 check points organized by the Swiss police in Bern, 61 drivers were tested for the presence of drugs of abuse using the Drugwipe 5 device. In parallel, oral fluid was collected with the Intercept DOA Oral Specimen Collection device and tested by gas chromatography-mass spectrometry (GC-MS) after methylation of THC (limit of quantitation 1 ng/mL). The Drugwipe device identified 1 exposed driver, but with GC-MS, 18 drivers tested positive. THC concentrations in the Intercept buffer ranged from 2.1 to 205.1 ng/mL. These concentrations represent about 1/2 to 1/3 the authentic THC concentrations in oral fluid because of the dilution by the blue liquid of the device. Two main limitations of oral fluid were 1. the amount of matrix collected is smaller when compared to urine and 2. the levels of drugs in urine are higher than in oral fluid. A current limitation of the use of this specimen for roadside testing is the absence of a suitable immunoassay that detects the parent compound in sufficiently low concentrations.

Advances in the use of mass spectral libraries for forensic toxicology.

Gas chromatography in combination with mass spectrometry (GC-MS) plays an important role in the field of analytical toxicology. The identification of unknown compounds is very frequently undertaken with GC-MS and utilizing mass spectral libraries. Currently available libraries for analytical toxicology were compared for overlapping and uniqueness of their entries. Furthermore, the widely known Pfleger-Maurer-Weber-Drugs-and-Pesticides-Library for toxicology (PMW_tox2) was used to compare the search algorithms PBM (Probability Based Matching, Agilent Technologies), INCOS (Finnigan/Thermoquest), and MassLib (Max Planck Institute). To our knowledge, direct comparisons of mass spectral libraries and search programs for analytical toxicology have not been published previously. The capabilities and necessities of modern MS technology in the field of general unknown analysis are revealed, and some of the potential pitfalls are described.