Band 37, Heft 4

Detection and quantification of synthetic cannabinoids (synonym: cannabimimetics) used as a substitute for natural cannabis has been a real toxicological and forensic issue since 2008. On the basis of a short overview of the pharmacological principle, chemical classification, and legal situation in Germany, the development of several analytical and screening approaches is presented. The paper further describes and validates a novel method for the simultaneous identification and quantification of JWH-018, JWH-019, JWH-073, JWH-081, JWH-122, JWH-200, JWH-210, JWH-250, WIN48,098, WIN55,212-2, AM-694 and CP47,497 by means of liquid chromatography-tandem mass spectrometry (LC-MS/MS) in human serum and hair using JWH-018-d 11 and THC-d 3 as internal standards. Detection limits ranging from 0.018 to 0.192 ng/mL for serum and from 0.140 to 0.820 pg/mg for hair, as well as very short retention times (up to 2.4 min), qualify this method, which presently includes 11 further compounds on a semi-quantitative basis, for rapid screening. The same method, using the deuterated 3-hydroxybutyl derivative of JWH-073, was applied for detection of 14 urinary metabolites after enzymatic hydrolysis of conjugates. Observation from routine analysis of 359 serum and 84 hair specimens reveals that concentrations of cannabimimetics in positive samples may differ by factors of at least 400 and 3000 with median values of 0.5 ng/mL and 9 pg/mg for serum and hair, respectively. Conclusions regarding consumption behaviors and strategies to decipher metabolic routes of these substances and possible implications are also discussed. Zusammenfassung: Der Nachweis und die Quantifizierung synthetischer Cannabinoide (Synonym: Cannabimimetika), die in Form von Räuchermischungen als Cannabisersatz in Gebrauch sind, ist seit 2008 ein aktuelles Thema der toxikologischen und forensischen Analytik. Auf der Grundlage einer kurzen Übersicht über das pharmakologische Prinzip, die chemische Klassifikation und die gesetzliche Situation in Deutschland wird die Entwicklung verschiedener Screeningmethoden dargestellt. Es wird eine neue Methode zur simultanen Quantifizierung von JWH-018, JWH-019, JWH-073, JWH-081, JWH-122, JWH-200, JWH-210, JWH-250, WIN48,098, WIN55,212-2, AM-694 und CP47,497 in Serum und Haaren mittels Flüssigchromatographie-Tandem-Massenspektrometrie (LC-MS/MS) mit JWH-018-d 11 und THC-d 3 als internen Standards beschrieben und validiert, die aufgrund der Nachweisgrenzen von 0.018 bis 0.192 ng/mL im Serum und 0.140 bis 0.820 pg/mg Haar sowie der besonders kurzen Retentionszeiten (maximal 2,4 Minuten) als Screeningmethode geeignet ist. Die Methode inkludiert derzeit 11 weitere Substanzen auf semiquantitativer Basis und ist nach enzymatischer Hydrolyse der Glucuronide unter Verwendung des deuterierten 3-Hydroxybutyl-Metaboliten von JWH-073 als internem Standard für die Suche nach 14 Urinmetaboliten verwendbar. Eine konsekutive Analytik der Routineanalytik von 359 Serum- und 84 Haarproben ergibt, dass die gemessenen Konzentrationen in positiven Proben um einen Faktor von 400 (Serum; Median 0.5 ng/mL) bzw. 3000 (Haare; Median 9 pg/mg) differieren können. Mögliche Rückschlüsse auf das Konsumverhalten und Ansätze zur Aufklärung von Stoffwechselrouten und ihre Bedeutung werden abschlieβend diskutiert.

Since the discovery of ANCA 30 years ago, there has been a substantial improvement in this detection technique. Current state of the art is indirect immunofluorescence (IFT) followed by antigen-specific assays. The performance of the available Proteinase 3- and Myeloperoxidase- (PR3- and MPO-) ANCA assays concerning sensitivity and specificity has been continuously improved, especially due to the use of capture and anchor ELISA techniques. However, the quality of several commercially available ANCA assays is low when tested with an international standard. The sole use of ELISA and abolition of IFT is not recommended. PR3- and MPO-ANCA have well-established significance in the diagnosis of ANCA-associated vasculitides. Despite the achieved increase in sensitivity and specificity of ANCA testing, increasing unselected and broad clinical use leads to a rise in “false” positive results. New detection methods, that might replace the currently used combination of IFT and ELISA, are under development but will need evaluation in international multicenter trials.

Zusammenfassung: Phäochromozytome, Paragangliome und Neuroblastome sind seltene neuroendokrine Tumoren, die durch eine erhöhte Katecholaminproduktion charakterisiert sind. Die Katecholamine werden noch in den Tumorzellen zu Metanephrinen O-methyliert. Diese Tatsache erklärt die hohe diagnostische Sensitivität und Spezifität der Katecholaminmetabolite zur Diagnostik von Phäochromozytomen und Paragangliomen. Altersangepasste Referenzwerte von Normetanephrin, dem O-methylierten Metabolit von Noradrenalin, erhöhen hierbei noch die diagnostische Spezifität. Gemessen an der höheren diagnostischen Sensitivität sind die auf Flüssigkeitschromatographie beruhenden Methoden, Metanephrine im Patientenplasma zu messen, immunologischen Verfahren überlegen. Für das Neuroblastom-Screening bleibt die Messung der Tumormarker Vanillinmandelsäure und Homovanillinsäure der am häufigsten benutzte Test, obwohl sie eine eingeschränkte diagnostische Sensitivität haben. Für die Diagnose solcher pädiatrischen Tumoren ist die Suche nach neuen, verbesserten Biomarkern notwendig. Abstract: Pheochromocytoma, paraganglioma, and neuroblastoma are rare neuroendocrine tumors characterized by elevated production of catecholamines. The catecholamines are O-methylated to metanephrines in tumor cells, explaining their high diagnostic sensitivity and specificity for diagnosis of pheochromocytoma and paraganglioma. Age-adjusted reference values of normetanephrine, the O-methylated metabolite of norepinephrine, are useful for improving diagnostic specificity. Among available methods for measuring plasma metanephrines, liquid chromatographic based analyses are the most accurate and precise. For screening of neuroblastoma, measurements of vanillylmandelic acid and homovanillic acid in urine remain the most commonly used tests, but have limited diagnostic sensitivity. For such childhood tumors there is a need for new and improved biomarkers for diagnosis.

Accelerated vascular calcification (VC) is one of the driving forces of high morbidity and mortality in patients with severe chronic kidney disease (CKD) or end-stage renal failure. VC is a hallmark of chronic cardiorenal syndrome and an integral part of CKD-mineral and bone disorder. Analyzing the pathogenesis of cardiovascular calcification in uremic vasculopathy has been the focus of research for years in nephrology. Unfortunately, many questions regarding prevention and treatment are still unanswered. Overall, VC is regarded as an actively regulated process. Numerous biomarkers have been defined, which play a role in the development of VC. Theoretically, laboratory parameters associated with ectopic calcification may serve as biomarkers, risk factors for VC, and/or monitoring tools regarding treatment effects. Interestingly, none of the currently known parameters are “perfect” in terms of optimal risk stratification, diagnosis, and management of patients with cardiorenal syndrome and VC. The aim of the present review is to give an overview on the current status of our understanding of uremic VC. Moreover, it presents, in detail, data on some selected biomarkers/risk factors in the field (fetuin-A, magnesium, alkaline phosphatase, matrix Gla protein). Well-substantiated data point towards the existence of a “bone-vascular axis” meaning that a reciprocal association exists between cardiovascular and bone disease in CKD patients, which most likely influence each other.

Background: Two months after a vacation in Thailand, a 54-year-old alcohol-addicted diabetes patient was admitted to hospital presenting with a febrile, therapy- refractory urinary tract infection. Burkholderia pseudomallei was cultured from urine but misidentified as Burkholderia cepacia . Application of inappropriate chemotherapy provoked systemic spread of bacteria from urinary tract abscesses. Septic shock and respiratory failure resulted in a fatal outcome. Results: Correct identification at the species level was achieved by 16S rRNA gene sequencing. Postmortem, B. pseudomallei was additionally detected in prostatic tissue by PCR and fluorescence in situ hybridization. Conclusions: Biochemical differentiation of bacteria of the genus Burkholderia is not reliable at the species level. Molecular approaches should be added if an infection with B. pseudomallei is probable according to clinical anamnesis. The recommended dosage of antibiotic drugs should be at the upper limit if abscess formation is likely.

Background: Two months after a vacation in Thailand, a 54-year-old alcohol-addicted diabetes patient was admitted to hospital presenting with a febrile, therapy-refractory urinary tract infection. Burkholderia pseudomallei was cultured from urine but misidentified as Burkholderia cepacia . Application of inappropriate chemotherapy provoked systemic spread of bacteria from urinary tract abscesses. Septic shock and respiratory failure resulted in a fatal outcome. Results: Correct identification at the species level was achieved by 16S rRNA gene sequencing. Postmortem, B. pseudomallei was additionally detected in prostatic tissue by PCR and fluorescence in situ hybridization. Conclusions: Biochemical differentiation of bacteria of the genus Burkholderia is not reliable at the species level. Molecular approaches should be added if an infection with B. pseudomallei is probable according to clinical anamnesis. The recommended dosage of antibiotic drugs should be at the upper limit if abscess formation is likely.