Werner Pogoda

Criminal poisoning of commuters in Bangladesh: Prospective and retrospective study

Travel-related poisoning is an emerging social and public health emergency in Bangladesh but its cause and significance have not been determined. To investigate this syndrome we performed a prospective clinical study and retrospective analysis of hospital records in a general medicine unit of a public tertiary care teaching hospital in Dhaka, Bangladesh, using toxicological analysis by fluorescence polarization immunoassay (FPIA) and liquid chromatography coupled to time-of-flight mass spectrometry (LC-TOF MS). The participants of the prospective study were 130 consecutive patients aged 16-80 years who were admitted with central nervous system depression (Glasgow Coma Score 3-14) after using public transportation, in the absence of other abnormalities, from January through June 2004, and a convenience sample of 15 such patients admitted during 3 days in May 2006. In 2004-2006, travel-related poisoning increased from 6.1 to 9.5% of all admissions (210-309 of 3266-3843 per year), representing 46.6-55.7% of all admitted poisoning cases. Incidents were associated with bus (76%), taxi, train, and air travel, or local markets; 98% of patients remembered buying or accepting food or drinks before losing consciousness. Direct financial damage (missing property) was diverse and frequently existential. Among 94 urine samples analyzed by FPIA, 74% tested positive for benzodiazepines. Among 15 urine samples analyzed by LC-TOF MS, lorazepam was detected in all; five also contained diazepam or metabolites; nitrazepam was present in three. FPIA results obtained for these 15 samples were below the recommended cut-off in eight (53%; lorazepam only). Our findings show that the massive medicosocial emergency of travel-related poisoning in Bangladesh is the result of drug-facilitated organized crime and that benzodiazepine drugs are used to commit these crimes, suggesting modifications to the local emergency management of the victims of this type of poisoning. They also highlight the need for more research in the neglected field of acute poisoning in Bangladesh, and for criminal investigations of the use of benzodiazepine drugs in this country.

Retrospective analysis of synthetic cannabinoids in serum samples – epidemiology and consumption patterns

Herbal mixtures contain synthetic cannabinoids, which can cause severe intoxications. Due to the great variety and the changing spectrum of substances on the drug market, prevalence data are limited, and data on prevalence rates of synthetic cannabinoids in forensic cases are not available. The present study was performed to survey the prevalence of synthetic cannabinoids in cases of traffic and criminal offences in the German state Hesse in 2010. The applied analytical method covered all synthetic cannabinoids on the drug market at that time, and with 20% of the blood samples (422 out of 2201) a representative number was reanalyzed. In twelve samples synthetic cannabinoids were identified and a prevalence of 2.8% was estimated. Consumption patterns showed predominantly cases of multi-drug consumption (10 cases); the combination with cannabis or alcohol was frequent (four cases each). The observed deficits were moderate with the exception of aggravation of paranoia in one case. The symptoms were either compatible with the effects of cannabinoid agonists or attributable to alcohol or other drugs found in the blood samples. Our current analytical strategy is to perform such analyses only in cases where use is suspected or where symptoms are not explained by routine toxicological analyses. Hence, the positive rate is rather low highlighting the need to keep up with the developments on the drug market and to establish sensitive screening methods covering a broad range of substances that can be updated fast, e.g., relying on collections of mass spectrometric reference data.

Pharmacokinetic properties of the synthetic cannabinoid JWH-018 and of its metabolites in serum after inhalation

&NA; Each year, synthetic cannabinoids are occurring in high numbers in the illicit drug market, but data on their pharmacology and toxicology are scarcely available. Therefore, a pilot study was performed to assess adverse effects of JWH‐018, which is one of the oldest and best known synthetic cannabinoids. Six subjects inhaled smoke from 2 and 3 mg JWH‐018. The drug and nine of its metabolites were analyzed in their blood samples taken during the following 12 h by liquid chromatography–mass spectrometry (LC–MSMS). The maximum concentration of JWH‐018 reached 2.9–9.9 ng/ml after inhalation and markedly decreased during the next 1.5 h, followed by a multiexponential decline (t1/2 in median 1.3 h and 5.7 h). The concentration of the pentanoic acid metabolite was slightly higher than that of the 3‐, 4‐ and 5‐hydroxypentyl metabolites and of the 6‐hydroxyindol metabolite. The data also suggest a multiexponential decline and slow terminal elimination of JWH‐018 and all metabolites. The detection of JWH‐018 and of its metabolites in serum requires high analytical sensitivity. The pharmacokinetic properties of inhaled JWH‐018 are similar to that of THC. A slow terminal elimination of drug and metabolites may lead to accumulation in chronic users. HighlightsAfter inhalation of JWH‐018, serum concentrations are maximal within minutes.During the 12 h period a multicompartmental distribution/elimination is obvious.Pentanoic acid, 3‐, 4‐ and 5‐hydroxypentyl and 6‐hydroxyindol met. are detected.Results indicate a novel metabolite, an isomer of the pentanoic acid metabolite.A slow terminal elimination may lead to accumulation in chronic users.

Poor alkaloid sequestration by arrow poison frogs of the genus Phyllobates from Costa Rica

Frogs of the genus Phyllobates from Colombia are known to contain the highly toxic alkaloid batrachotoxin, but species from Central America exhibit only very low levels or are entirely free of this toxin. In the present study alcohol extracts from 101 specimens of Phyllobates lugubris and Phyllobates vittatus and 21 of three sympatric species (Dendrobates pumilio, Dendrobates auratus, Dendrobates granuliferus) from Costa Rica were analyzed by gas chromatography-mass spectrometry. Whereas the extracts of the Dendrobates species exhibited typical profiles of toxic alkaloids, those of the two Phyllobates species contained low levels of few alkaloids only, batrachotoxin was not detected. Although the feeding pattern of the Dendrobates and Phyllobates species are similar as revealed by examination of their stomach content (mainly ants and mites), the Phyllobates species are poorly sequestering alkaloids from their food source in contrast to the Dendrobates frogs.

Estimating the age of the adult stages of the blow flies Lucilia sericata and Calliphora vicina (Diptera: Calliphoridae) by means of the cuticular hydrocarbon n-pentacosane

Age estimation of insects like blow flies plays an important role in forensic entomology and can answer questions in regard to time of death. So far the focus is on the immature stages of these insects, but recently the adult fly became a target of interest. It has been established that the profile of specific cuticular hydrocarbons (CHCs) changes in a consistent pattern as adult insects age; thus, their analysis could be a promising tool for the age estimation of adult insects. We investigated the CHC n-pentacosane (nC25) on the legs of the adult blow flies Lucilia sericata and Calliphora vicina with gas chromatography-mass spectrometry. The flies were kept at room temperature (17°C±2°C) and 12:12 L:D from Day 1 to Day 20 post-emergence. For each of five flies per species, the amount of nC25 on all legs was determined daily. The amounts of nC25 on C. vicina increased linearly (R2=0.949). No significant difference between sexes could be detected. While L. sericata showed the same linear increase in general, we found significant (p<0.001) differences in the amount of nC25 between males and females. Although the amounts of nC25 increased linearly for both sexes (males: R2=0.948; females: R2=0.920), female L. sericata produced more nC25 than males. An equation for the prediction of fly age is constructed from these data. Although the influence of various environmental factors, e.g., fluctuating temperatures, still needs to be tested, nC25 seems to be a promising tool for the age estimation of adult flies.

Excretion of metabolites of the synthetic cannabinoid JWH-018 in urine after controlled inhalation

Each year, synthetic cannabinoids are occurring in high numbers on the illicit drug market but data obtained after controlled application are rare. The present study on pharmacokinetics in urine is part of a pilot study on adverse effects of JWH-018, which is one of the oldest and best known synthetic cannabinoids. Six subjects inhaled smoke from 2 and 3mg JWH-018. The drug and ten potential metabolites were analyzed in urine samples collected during 12h after inhalation by liquid chromatography-mass spectrometry (LC-MS/MS) without and with conjugate cleavage. The parent compound was not detectable, but 13 of its metabolites, all of which were conjugated. Concentrations of the predominant metabolite, JWH-018 pentanoic acid, were less than 5ng/ml, but in two subjects it was still detected up to 4 weeks after ingestion. Other major metabolites were 5- and 4-HOpentyl-JWH-018, JWH-073 butanoic acid and a hypothetically dihydroxylated and dehydrogenated metabolite of JWH-018. Occasionally, further hydroxylated metabolites were found. Generally, hydroxylated metabolites were detected in concentrations lower than 1ng/ml already 10h after inhalation. All concentrations were much lower than reported for urine samples of authentic JWH-018 users. The formation of the metabolite JWH-018 pentanoic acid was found to be slightly delayed, but its rather high concentrations and detection over several weeks after single dosing makes it a useful target for urine analysis. The different excretion of carboxylic acid and hydroxylated metabolites may aid in evaluation of time of use.

Pharmacokinetic properties of the synthetic cannabinoid JWH-018 in oral fluid after inhalation

Each year, synthetic cannabinoids occur in high numbers on the illicit drug market, but data on their detectability are rarely available. A pilot study was performed to assess adverse effects of JWH-018, which is one of the oldest and best known synthetic cannabinoids. Oral fluid has been evaluated as a specimen for drug monitoring. Six subjects inhaled smoke derived from 2 and 3 mg JWH-018. The drug and 10 of its metabolites were analyzed in oral fluid samples collected during the following 12 hours using the Quantisal collection device by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Maximum concentrations of JWH-018 reached 2.2-2036 (median 25.7) ng/mL after inhalation and decreased during the next hour to only 0.08-8.42 (median 0.89) ng/mL. Metabolites were not found. During the elimination phase (median half-life 1.69 hours), detection of the drug over 6-12 hours (median 8 hours) after inhalation was achieved (0.024 ng/mL limit of quantification). Oral fluid/serum ratios varied considerably intra- and inter-individually in a range of 0.05-555 (median 1.38). The detection of JWH-018 in oral fluid requires high analytical sensitivity even 1 hour after inhalation. The pharmacokinetic properties of inhaled JWH-018 are similar to those of THC. Times for detection are typically less than 12 hours. High variability of the oral fluid/serum ratio precludes extrapolation of oral fluid concentrations to blood.

Feeding on toxic prey. The praying mantis (Mantodea) as predator of poisonous butterfly and moth (Lepidoptera) caterpillars

ABSTRACT Caterpillars of the monarch butterfly, Danaus plexippus, feed on milkweed plants, Asclepias spp. (Apocynaceae), and sequester their toxic cardenolides aimed at deterring predators. Nevertheless, Chinese praying mantids, Tenodera sinensis, consume these caterpillars after removing the midgut (“gutting”) including its plant content. In the present study, monarch caterpillars raised on A. curassavica, and those of the deaths‐head hawkmoth, Acherontia atropos, raised on Atropa belladonna containing atropine, were fed to mantids, Hierodula membranacea, which removed the gut of both species discarding about 59% of cardenolides and more than 90% of atropine, respectively. The ingestion of these compounds produced no apparent ill effects in the mantids and both were excreted with faeces. On the other hand, when mantids were fed with larvae of two moth species, Amata mogadorensis and Brahmaea certia, raised on non‐poisonous host plants, the mantids showed the same gutting behaviour, thereby discarding indigestible plant material. As polar compounds, e.g. cardenolides and atropine, are not absorbed from the mantids midgut and do not pass the gut membrane, this enables the mantids to feed on toxic prey. HIGHLIGHTSThe praying mantis feeds on toxic caterpillars and discards the intestines.However, the mantis still ingests toxins, which are secreted later with faeces.But the mantis shows the same behaviour when feeding on non‐toxic caterpillars.To get rid of plant material in the intestines seems to be the main reason.

Loss of skin alkaloids in poison toads, Melanophryniscus klappenbachi (Anura: Bufonidae) when fed alkaloid-free diet

Toads of the genus Melanophryniscus contain toxic alkaloids sequestered from a diet of mainly ants and mites. Wild-caught specimens of M. klappenbachi were fed an alkaloid-free diet and their alkaloid profile was analyzed during 38 months in captivity. Individual alkaloid patterns varied considerably. Over time, the concentration of two alkaloids, pumiliotoxin 251D and 3,5-disubstituted indolizidine 195B, significantly declined, suggesting that the toads depend on continuous access to alkaloid-containing prey to maintain natural levels of their chemical defense.

Lack of alkaloids and tetrodotoxin in the neotropical frogs Allobates spp. (Aromobatidae) and Silverstoneia flotator (Dendrobatidae)

Frogs of the family Dendrobatidae are known to contain toxic alkaloids in their skin secretion, but Allobates species of the closely related Aromobatidae family are considered to lack toxic secretions. However, contradictory results have been reported. Analyses of alcohol extracts from three different Allobates species from South-America (Guiana Shield), Central America (Costa Rica), and from the dendrobatid frog Silverstoneia flotator confirm the absence of alkaloids and tetrodotoxin in aromobatids and in a dendrobatid of the subfamily Colostethinae.