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In contrast to cumulative techniques (e.g., tape-lift) for qualitative gunshot residues (GSR) analysis, topographic methods are commonly applied to preserve the integrity of evidence from a shooter's or victim's hand in cases of gun-related crimes. Topographic sampling techniques employing adhesive foils, latex, or the polyvinyl alcohol (PVAL) method enable unambiguous sampling of biological and non-biological trace material while preserving its spatial distribution and relation to each other. The PVAL method in particular allows for a topographically veridic and quantitative conservation of traces of GSR and biological stains that are embedded in the PVAL glove, because it completely removes these traces from the hand. The present study investigated the success rates of STR profiling and the detection of blood and brain-specific gene expression from minimal traces of blood splatter as well as parallel to the positive detection of gunshot residues embedded in 17 PVAL gloves taken from the hands of deceased persons in the context of homicide cases in the period between 1996 and 2003. The water-soluble PVAL matrix is shown to be fully compatible with successful STR profiling and the detection of blood- and brain-specific miRNA expression, even after up to 20 years of storage, demonstrating that this sampling technique offers advantages compared to other more simplistic sampling methods like taping.

Confirmation or exclusion of recent heroin consumption is still one of the major challenges for forensic and clinical toxicologists. A great variety of biomarkers is available for heroin abuse confirmation, including various opium alkaloids (eg, morphine, codeine), street heroin impurities (eg, 6-acetylcodeine [6-AC], noscapine, papaverine) as well as associated metabolites (eg, 6-monoacetylmorphine [6-MAM], morphine glucuronides). However, the presence of most of these biomarkers cannot solely be attributed to a previous heroin administration but can, among other things, also be due to consumption of poppy seed products ('poppy seed defense'), opium preparations or specific medications, respectively. A reliable allocation is of great importance in different contexts, for instance in the case of DUID (driving under the influence of drugs) investigations, in driving licence re-granting processes, in workplace drug testing (WDT), as well as in post-mortem identification of illicit opiate use. Additionally, differentiation between illicit street heroin abuse and pharmaceutical heroin administration is also important, especially within the frame of heroin-assisted treatments. Therefore, analysis of multiple biomarkers is recommended when illicit opiate consumption is assumed to obtain the most reliable results possible. Beyond that, interpretation of positive opiate test results requires a profound insight into the great variety of biomarkers available and their validity regarding the alleged consumption. This paper aims to provide an overview of the wide variety of heroin abuse biomarkers described in the literature and to review them regarding their utility and reliability in daily routine analysis., (Copyright © 2017 John Wiley & Sons, Ltd.)

Purpose: In the present study we characterized a series of synthetic cannabinoids containing various heterocyclic scaffolds that had been identified as constituents of "Spice", a preparation sold on the illicit drug market. All compounds were further investigated as potential ligands of the orphan receptors GPR18 and GPR55 that interact with some cannabinoids., Methods: The compounds were studied in radioligand binding assays to determine their affinity for human cannabinoid CB 1 and CB 2 receptors expressed in CHO cells, and in cAMP accumulation assays to study their functionality., Results: Structure-activity relationships were analyzed. The most potent CB 1 receptor agonist of the present series MDMB-FUBINACA ( 12 ) ( K i  = 98.5 pM) was docked into the human CB 1 receptor structure, and a plausible binding mode was identified showing high similarity with that of the co-crystallized THC derivatives. MDMB-CHMCZCA ( 41 ) displayed a unique profile acting as a full agonist at the CB 1 receptor subtype, but blocking the CB 2 receptor completely. Only a few weakly potent antagonists of GPR18 and GPR55 were identified, and thus all compounds showed high CB receptor selectivity, mostly interacting with both subtypes, CB 1 and CB 2 ., Conclusions: These results will be useful to assess the compounds' toxicological risks and to guide legislation. Further studies on 41 are warranted., Competing Interests: Compliance of ethical standardsThe authors declare that they have no conflict of interest.This article does not contain any studies with human participants or animals performed by any of the authors.

Besides its clinical application, the anaesthetic agent propofol is being increasingly misused, mostly by healthcare professionals, and its abuse potential gained worldwide attention after the tragic death of Michael Jackson in 2009. Due to the short duration of its narcotic effects, propofol abuse is especially easy to hide compared with the use of other recreational drugs. However, propofol possesses a very narrow therapeutic window between the desired effect and potentially fatal toxicity, making abuse of the drug extremely dangerous even in experienced physicians. Consequently, it is important that forensic laboratories possess a sensitive and specific method for the detection of chronic propofol abuse. We present a simple, fast and reliable method to simultaneously extract propofol and its main metabolite propofol glucuronide from hair, followed by sensitive LC-MS/MS analyses, allowing to determine a chronic propofol abuse. Difficulties regarding the detection of propofol using LC-MS/MS were solved by using a derivatization reaction with 2-fluoro-1-methylpyridinium-p-toluene-sulfonate and triethylamine. Reliability of extraction method and subsequent LC-MS/MS analyses was confirmed under consideration of the validation parameters selectivity, linearity, accuracy and precision, analytical limits, processed sample stability, matrix effects and recovery. Appropriate quantification (LLOQ=10pg/mg hair) and detection limits (3.6pg/mg hair for propofol and 7.8 pg/mg hair for propofol glucuronide) could be achieved, enabling to detect even small amounts of both analytes. Applicability of the method was confirmed by analysis of three human hair samples from deceased with suspicion of chronic propofol abuse., (Copyright © 2017 Elsevier B.V. All rights reserved.)

The expression of heat shock proteins (hsps) increases in cases of hyperthermal cellular stress in order to protect cellular structures. Hsps can be visualized with immunohistochemical staining. We examined 48 cases of death from fire and excessive heat and a control group of 100 deaths without any perimortem thermal stress, measuring both the hsp27 and hsp70 expressions in myocardial, pulmonary, and renal tissues. The results revealed a correlation between hsp expression and survival time. Hsps are expressed rapidly within seconds or minutes after exposure to heat stress. In particular, hsp27 is expressed fast in high levels, whereas hsp70 expression is higher in the pulmonary and renal tissue of long-term survivors. In the myocardial tissue, hsp27 expression dominated in both short- and long-term survival. The expression pattern is strongly dependent on the organ structure and the survival time, which should be considered in future postmortem studies on hsps.

Metabolism of the tricyclic azaphenothiazine neuroleptic drug prothipendyl was investigated with in vitro studies using human liver microsomes but also specific isoforms of cytochrome P450 (CYP) enzymes. Identification and analysis of metabolites was done by liquid chromatography (LC) coupled with quadrupole time of flight mass spectrometry (LC-QTOF-MS) as well as triple quadrupole mass spectrometry (LC-QQQ-MS). Results of the herein presented study revealed the proof of various demethylated and oxidized metabolites (-CH 2 , -C 2 H 4 , four derivatives of prothipendyl +O and three derivatives of prothipendyl -CH 2 +O). Metabolic reactions of prothipendyl were mainly catalyzed by CYP enzymes CYP1A2, CYP2D6, CYP2C19 and CYP3A4. N-demethyl-prothipendyl was predominantly formed by isoforms CYP2C19 and CYP1A2, while particularly the CYP isoenzyme 3A4 was responsible for the formation of prothipendyl sulfoxide. To confirm the formation of previously identified metabolites in vivo, cardiac blood samples that were tested positive for prothipendyl during routine drug testing and serum and urine samples, collected after a voluntary intake of prothipendyl, were analyzed by LC-QQQ-MS. All metabolites of prothipendyl were proven in these authentic specimens. Neither in serum samples nor in urine samples, a prolonged detectability of metabolites in comparison to prothipendyl could be demonstrated., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Recently, phase-II-metabolites of γ-hydroxybutyric acid (GHB), namely GHB-β-O-glucuronide and GHB-4-sulfate, were implemented in the scope of drug testing methods The clearance of GHB from the circulation is extremely fast due to its incorporation into the metabolic pathway of the citrate cycle. The elimination half-life of GHB from blood was reported to be dose dependent between 30 and 50min resulting in narrow detection windows of less than 12h after illicit administration or cases of drug facilitated sexual assault regardless of the biological matrix used. As sulfated metabolites tend to show prolonged half-lives and slower elimination kinetics compared to unmodified or glucuronidated drugs, the potential of GHB-4-sulfate in prolonging the detection of GHB administration was assessed. Its urinary concentrations were determined in n=100 samples from athletes and n=50 samples from sport students, and the resulting data were used to calculate a preliminary reference population-based threshold for urinary GHB-sulfate concentration. The threshold was then compared to concentrations found in post-administration urine samples collected from 3 volunteers who administered GHB within the setting of a clinical trial. Due to the large inter-individual variability of concentrations found in the reference population, GHB-4-sulfate itself was not suitable to prolong the detection times for GHB applications, even when specific gravity-corrected values were used. Therefore, a metabolomics-based approach was applied to the reference population samples and evaluated regarding other urinary metabolites that potentially correlate with the urinary excretion of GHB-4-sulfate and GHB-β-O-glucuronide in order to find a suitable marker to normalize urinary concentrations. The most promising candidate was found at a molecular mass of 321.0696 and was preliminarily identified as β-citryl-glutamic acid., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Information about stability of synthetic cannabinoids is important to give recommendations for storage conditions in cases in which use of synthetic cannabinoids is suspected. In this study, freeze-thaw stability (3 cycles at -20 °C at 1.5 ng/mL) and long-term stability at room temperature, 4°C and -20°C (for 150 days) were tested by a validated liquid chromatographic-mass spectrometric method for 80 synthetic cannabinoids. Results demonstrated good freeze-thaw stability for most of the substances. For 5 F-ABICA (72.6% of peak area of time point zero), 5 F- 2 ADB PINACA 2 Isomer (68.7 %), 5Cl-AB-PINACA (84.2 %), AB FUBINACA 2-Isomer (33.5 %), 5 F-PCN (89.7 %), ADB-FUBINACA (78.0 %), EG 018 (88.9 %), and PX-1 (89.4 %) three freeze-thaw cycles led to absolute peak areas < 90% compared to 6 control samples. For 51 of the 84 substances, stability in serum could be demonstrated after 315 days of storage at -20°C. For 82 of the 84 substances, stability in serum could be shown for at least 1 month at -20°C. Long- term stability at 4°C or 20°C showed worse results. Therefore, it is our recommendation to store real serum samples at -20°C until analysis. Copyright © 2016 John Wiley & Sons, Ltd., (Copyright © 2016 John Wiley & Sons, Ltd.)

Gamma-hydroxybutyric acid (GHB) can be used as a knock-out drug in drug facilitated crime (DFC). Due to its rapid metabolism and resulting narrow detection window, uncovering GHB use in DFC still constitutes a problem. In this experiment we determined the GHB and GHB-β-O-glucuronide (GHB-Gluc) concentrations in hair samples after single and chronic GHB exposures. Hair samples of three narcoleptic patients therapeutically taking sodium oxybate (GHB-sodium-salt) were collected. Patients 1 (P1) and 2 (P2) took the medication for nine and six years, respectively. P1 took daily the pharmaceutical Xyrem ® in a total dose of 5.78g GHB at bed time (2.89g) and four hours (2.89g) later. P2 took a dose of 3.10g GHB at bed time and an additional dose of 2.68g GHB four hours later. Patient 3 (P3) was newly diagnosed with narcolepsy and started his therapy with oral dose of 6g (divided in three portions of 2g GHB) within 24h. The hair samples were extracted both with and without forerunning washing steps. GHB and GHB-Gluc were determined by a published ultra-high performance liquid chromatography-tandem mass spectrometry method using GHB-d 6 and GHB-Gluc-d 4 as internal standards. GHB and GHB-Gluc concentrations in unwashed hair samples of P1 and P2 were determined in a range of 0.56-1.30ng/mg and <0.48-0.85ng/mg, respectively. In washed hair samples of P1 and P2 the concentrations were in a range of <0.32-0.68ng/mg and <0.48-1.20ng/mg for GHB and GHB-Gluc, respectively. The determined concentrations were within the published endogenous range. The confirmed results showed that the washing procedure before extraction causes a minor decrease of GHB concentrations in hair (difference: <1ng/mg). The investigations showed that a single GHB exposure might not be determined by hair analysis of GHB and GHB-Gluc. The chronical intake of therapeutic sodium oxybate with doses up to 7g per night was also not confirmed by hair analysis maybe due to hair treatments. Therefore, GHB hair analysis should be assessed critically and determined negative results could not exclude GHB exposures., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Gamma-hydroxybutyric acid (GHB) acts as an agonist of the GABA B receptor, where GHB induces a depressant effect in the central nervous system. Besides its therapeutic application, GHB is also used as a date rape drug. However, the detection of GHB ingestion proves to be difficult due to its narrow detection window. The aim of this pilot study was to assess differential gene expressions after GHB intake to identify potential biomarkers for the detection of GHB intake. To this aim, alteration in gene expression of ALDH5A1, AKR7A2, EREG, and PEA15 was investigated via quantitative PCR (qPCR). Data normalization was based on a previously established and empirically derived normalization strategy. Blood samples of patients (n = 3) therapeutically taking sodium oxybate solution (GHB) and of donors without GHB intake (n = 49) were analyzed and compared. All qPCR procedures and results are reported according to the MIQE guidelines. Investigation of suitable reference genes using established algorithms suggested PPIB and FPGS as best-suited normalizers. Alterations in gene expression relating to GHB intake could not be confirmed to a forensically sufficient degree. However, significant differences in expression of EREG in the control group were observed, when time-point of sample collection was considered, indicating circadian rhythm. The study's main limitation is the small number of study subjects. Herein, we are first to present an empirically derived strategy for a robust normalization of qPCR data from the analysis of GHB-induced gene expression in human blood. We present results of the analysis of differential expression of ALDH5A1, AKR7A2, EREG, and PEA15 in the GHB-negative population. Finally, we report our findings on the effect of GHB intake on the expression of these genes and their presumable potential as GHB biomarkers.

Hypothermia causes systemic cellular stress. The pituitary gland is an endocrine gland and plays an important role in thermoregulation. When the core body temperature drops, the pituitary gland is activated by stimulation of hypothalamic hormones. In this study, we investigated morphological alterations of the pituitary gland in cases of fatal hypothermia. Several morphological alterations of the anterior lobe of the pituitary gland, such as hemorrhage, vacuolization, and hyperemia, have been previously described in fatal hypothermia. However, the diagnostic value of these findings is controversial. We compared 11 cases of fatal hypothermia with 10 cases lacking antemortem hypothermic influences. In the presence of thermal cellular stress, the expression of heat shock proteins increases to protect cellular structures. Therefore, we immunohistochemically analyzed Hsp27 and Hsp70. Hsp27 expression was detected in 27.3% of the cases of fatal hypothermia and in 10.0% of the control cases, whereas Hsp70 expression was not detected in any case. Additionally, Sudan staining was performed to quantify fatty degeneration. A positive reaction was found in 45.5% of the study group and in 10.0% of the control group. This indicates that fatty degeneration might be a valuable marker when other macroscopic signs of hypothermia are absent.

In cases of death caused by hypothermia, histological analysis can be used to determine the cause of death. Certain histological alterations of the pituitary glands in hypothermia have been reported in the literature, including haemorrhage, hyperaemia and cellular vacuolization of cells in the anterior lobe. In the present study, the validity of these morphological alterations as markers for fatal accidental hypothermia was investigated in autopsy material. A total of 34 pituitary glands in cases of verified fatal accidental hypothermia were examined histologically (haematoxylin and eosin, ferric, azan) and immunohistochemically (LCA, ACTH, C5b-9). The findings were compared with 61 cases in a control group. Hyperaemia was found in 50.0% of the study group cases and 59.0% of the control group cases. Cellular vacuolization was observed in one case (2.9%) in the study group and one case (1.6%) in the control group. Acute or recent haemorrhage in the glandular tissue was never detected. In our study, the histopathological characteristics described in the literature as pathognomonic for hypothermia could not be confirmed. Furthermore, histological differences in the pituitary glands between fatal hypothermia cases and control group cases were not observed.

In cases of firearm related fatalities a systematic investigation at the scene of death is indispensable to differentiate between self-inflicted and homicidal gunshot injuries. A common method to preserve gunshot residues (GSR) is their collection using adhesive tapes. However, the biological material gathered at the same time by the tapes would be of special interest if backspatter, ejected from the entrance wound against the direction of fire, could be detected. In the present study we examined the success rate of co-analysis of RNA and DNA recovered from biological traces sampled with adhesive tapes. The material originated from eight cases of fatal gunshots, taken from the hands of suspects or victims, examined 5 to 19 years ago for GSR. For all types of adhesive tapes tested, quantity and quality of the co-extracted nucleic acids was insufficient for successful DNA profiling, but was sufficient for the detection of blood-specific micro RNA (miRNA). In summary, sampling trace evidence from the hands of persons involved in fatal gunshots with adhesive tapes has a long-term detrimental effect on biological traces.

The Internet is flooded with steadily changing synthetic cannabinoids in `Spice` products. In routine forensic work, it is difficult to keep the analytical methods for the detection of these analytes up to date. We describe a liquid chromatography-tandem mass spectrometry method after liquid-liquid extraction for the detection of 93 synthetic cannabinoids in human serum. The method was validated for selectivity and specificity, matrix effects, and analytical limits (<1 ng/mL for 81 substances) for qualitative analysis. A short quantitative validation regarding linearity and precision data was also conducted. The method was applied to 189 serum samples provided by police authorities. Sixty-four samples (33.8%) were found positive for at least one synthetic cannabinoid, whereby MDMB-CHMICA, AB-CHMINACA, and 5 F-PB-22 were the substances most frequently detected. Consumption of these substances and plasma concentrations are linked to symptoms documented by the police. Six case reports are presented. Copyright © 2016 John Wiley & Sons, Ltd., (Copyright © 2016 John Wiley & Sons, Ltd.)

Separation and identification of positional isomers is an important issue in forensic toxicology, particularly in the context of new psychoactive substances (NPS). Despite the structural similarity, positional isomers often show different pharmacological properties and thus can exhibit dramatic differences with respect to their toxicity. Additionally, besides these pharmacological and toxicological effects, the legal status is also of great importance. We present a sensitive and selective LC-MS/MS method to separate the ortho, meta and para isomers of methylmethcathinone (MMC) and methylethcathinone (MEC) using a core-shell biphenyl analytical column. Reliability of the method was confirmed under consideration of the validation parameters selectivity, linearity, accuracy and precision, analytical limits, processed sample stability, matrix effects and recovery. Linearity was demonstrated over the entire calibration range from 5 to 250ng/ml with the use of a 1/x 2 weighting. Appropriate quantification and detection limits (LLOQ=5ng/ml, LOD<2ng/ml) could be achieved. Application of the method to real serum samples collected between June 2014 and August 2016 revealed the proof of a recent MMC or MEC consumption, respectively, in eight cases. Isomers of MMC could be detected in three of these eight cases, of which two were positive for 3-MMC and one was positive for 2-MMC. The other samples were tested positively for 3-MEC. In none of the samples 4-MMC, 2-MEC or 4-MEC could be detected. Only substances that were not governmentally controlled at that time could be detected, reflecting the rapid response of the recreational drug market to newly enacting drug laws., (Copyright © 2017 Elsevier B.V. All rights reserved.)

After contact shots to the head biological traces inside firearm barrels can be found. This study was conducted to simulate and to evaluate such staining. Five current handguns of four inch barrel length in the calibre .22 long rifle, 7.65mm Browning, 9mm Luger and .38 special were used to perform 24 contact shots on silicone coated, gelatine filled box models using the triple contrast method. The staining was documented by endoscopy and swabs gathered from both ends of the barrel were analysed by quantitative PCR. With the exception of the .22 revolver, all firearms showed distinct staining which decreased from the muzzle to the rear end of the barrel. The pattern was varied, showing droplets, elongated forms or stripes. In 14 of 24 shots, staining reached the chamber. The staining results were comparable to real suicide cases., (Copyright © 2017 Elsevier B.V. All rights reserved.)

For the estimation of time since death in the early postmortem interval, a large repertoire of valid methods is available. With increasing time since death the estimation of the postmortem interval becomes more imprecise and unreliable because validated methods are missing. Wehner et al. developed a method to estimate the time since death using immunohistochemical staining. Proteins undergo degradation after death and this reduces their stainability. Previous studies have explored the development of a method to estimate the time since death using immunohistochemical staining. A systematic analysis (Wehner et al., 1999-2002) demonstrated that calcitonin can still be stained 4 days, thyreoglobulin 5 days, glucagon 6 days, and insulin 12 days after death. After 12 days, calcitonin and thyroglobulin can no longer be stained, glucagon after 14 days, and insulin after 29 days. The aim of the present study was to test this original data on independent case material. Included in this control study were 105 cases with known time since death (between several hours and 22 days). Pancreatic tissue was stained for insulin and glucagon, and the thyroid gland for thyreoglobulin and calcitonin. The original findings could be generally confirmed, however for calcitonin and thyreoglobulin we observed earlier negative stainings. Altogether immunohistochemistry may still be useful as an additional method for estimating time since death in forensic cases., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Coronary bypass grafting is a routine procedure in heart surgery, which can now also be performed using a minimally invasive technique on the beating heart. In the presented case, a 55-year-old man died two days after bypass surgery. His condition had deteriorated post-operatively over 2 days until he finally died from multi-organ failure and cardiogenic shock. Autopsy revealed a fatal haemorrhage into the chest cavities from a small tear in the bypass and acute myocardial infarction. The haemorrhage had not been noticed in the hospital. Postoperative haemorrhage is a common complication after this type of surgery, which occurs in about 1-2 % of cases.

Cases in which several persons who died from an unnatural cause are found together are often difficult. It is necessary to exclude homicide committed by another person and to clarify whether the deaths are the result of a homicide-suicide or a joint suicide of persons wishing to die. Two cases in which couples with gunshot wounds to the head had been found lifeless in their homes are presented. In both cases, the deceased were of advanced ages and suffered from severe pre-existing diseases. Due to the circumstances at the scene, the results of the investigations and the autopsies as well as the suicide notes found, a double suicide was assumed in both cases. The husbands killed themselves after shooting their wives. Based on the presented cases the so-called double suicide and the need for a thorough investigation of the death scene with the problem of differentiating it from homicide-suicide and double homicide are discussed.

A male driver was checked during a traffic stop. A blood sample was collected 35min later and contained 7.3ng/mL THC, 3.5ng/mL 11-hydroxy-THC and 44.6ng/mL 11-nor-9-carboxy-THC. The subject claimed to have used two commercially produced products topically that contained 1.7ng and 102ng THC per mg, respectively. In an experiment, three volunteers (25, 26 and 34 years) applied both types of salves over a period of 3days every 2-4h. The application was extensive (50-100cm 2 ). Each volunteer applied the products to different parts of the body (neck, arm/leg and trunk, respectively). After the first application blood and urine samples of the participants were taken every 2-4h until 15h after the last application (overall n=10 urine and n=10 blood samples, respectively, for each participant). All of these blood and urine samples were tested negative for THC, 11-hydroxy-THC and 11-nor-9-carboxy-THC by a GC-MS method (LoD (THC)=0.40ng/mL; LoD (11-hydroxy-THC)=0.28ng/mL; LoD (THC-COOH)=1.6ng/mL;. LoD (THC-COOH in urine)=1.2ng/mL). According to our studies and further literature research on in vitro testing of transdermal uptake of THC, the exclusive application of (these two) topically applied products did not produce cannabinoid findings in blood or urine., (Copyright © 2017 Elsevier Ireland Ltd. All rights reserved.)

Complex suicides represent only a small number of all suicides. The case of a 27-year-old man who drove his car against a tree is presented. The emergency physician diagnosed a stab to the thorax. A kitchen knife was found on the rear seats of the vehicle. Autopsy revealed death by exsanguination due to perforation of the left cardiac ventricle. Blunt force injuries caused by the collision were also found.

Background: Propofol (2,6-diisopropylphenol) is a water-insoluble, intravenous anesthetic that is widely used for the induction and maintenance of anesthesia as well as for endoscopic and pediatric sedation. After admission, propofol undergoes extensive hepatic and extrahepatic metabolism, including direct conjugation to propofol glucuronide and hydroxylation to 2,6-diisopropyl-1,4-quinol. The latter substance subsequently undergoes phase II metabolism, resulting in the formation of further metabolites (1quinolglucuronide, 4quinolglucuronide and 4quinol-sulfate). Further minor phase I propofol metabolites (2-(ω-propanol)-6-isopropylphenol and 2-(ω-propanol)-6-isopropyl-1,4-quinol)) are also described. Due to its chemical structure with the phenolic hydroxyl group, propofol is also an appropriate substrate for sulfation by sulfotransferases., Methods: The existence of propofol sulfate was investigated by liquid chromatography electrospray ionization triple quadrupole mass spectrometry (LCESIQQQ-MS) and liquid chromatography electrospray ionization quadrupole time-of-flight mass spectrometry (LCESI-QTOF-MS). A propofol sulfate reference standard was used for identification and method development, yielding a precursor at m/z 257 (deprotonated propofol sulfate) and product ions at m/z 177 (deprotonated propofol) and m/z 80 ([SO3]-)., Results: Propofol sulfate - a further phase II metabolite of propofol - was verified in urine samples by LC-ESI-QQQ-MS and LC-ESI-QTOF-MS. Analyses of urine samples from five volunteers collected before and after propofol-induced sedation verified the presence of propofol sulfate in urine following propofol administration, whereas ascertained concentrations of this metabolite were significantly lower compared with detected propofol glucuronide concentrations., Conclusions: The existence of propofol sulfate as a further phase II propofol metabolite in humans could be verified by two different detection techniques (LCESIQQQ-MS and LC-ESI-QTOFMS) on the basis of a propofol sulfate reference standard. Evaluation of the quantitative analyses of propofol sulfate imply that propofol sulfate represents a minor metabolite of propofol and is only slightly involved in human propofol clearance.

Discrimination between street heroin consumption and poppy seed ingestion represents a major toxicological challenge in daily routine work. Several difficulties associated with conventional street heroin markers originate from their versatile occurrence in various poppy seed products and medications, respectively, as well as to small windows of detection. A novel opportunity to overcome these hindrances is represented by the new potential street heroin marker acetylated-thebaine-4-metabolite glucuronide (ATM4G), originating from thebaine during street heroin synthesis followed by metabolic reactions after administration. In this study, urine samples after consumption of different German poppy seed products and urine samples from subjects with suspicion of preceding heroin consumption were tested for ATM4G, 6-AC (6-acetylcodeine), papaverine, noscapine, 6-MAM (6-monoacetylmorphine), morphine, and codeine. Neither 6-AC and 6-MAM nor ATM4G but morphine and codeine could be detected in urine samples following poppy seed ingestion. As well, neither papaverine nor noscapine could be observed even after consumption of poppy seeds containing up to 37 µg noscapine and up to 9.8 µg papaverine, respectively. Concerning the urine samples with suspicion of preceding heroin consumption, ATM4G could be detected in 9 of 43 cases. By contrast, evidence of 6-AC and 6-MAM, respectively, could only be seen in 7 urine samples. In conclusion, ATM4G should be measured additionally in cases requiring discrimination of street heroin consumption from poppy seed intake. Copyright © 2016 John Wiley & Sons, Ltd., (Copyright © 2016 John Wiley & Sons, Ltd.)

Analysis of the anesthetic agent propofol in biological samples by LC-MS/MS is a great challenge due to weak fragmentation and poor ionization efficacy of propofol resulting in weak signal intensities. Improvements of the ionization and fragmentation efficacy can be achieved by conversion of propofol to its dimethylimidazolesulfonyl (DMIS) derivative by a derivatization reaction using 1,2-dimethylimidazole-4-sulfonyl chloride (DMISC). This DMIS derivative produced intense [M + H] + ions in positive-ion LC-ESI-MS/MS with the dimethylimidazole moieties representing the most abundant product ions. Derivatization of serum samples is achieved by direct conversion of the acetonitrile supernatant of a protein precipitation with DMISC followed by a double liquid-liquid extraction using n-hexane. Reliability of the method was confirmed under consideration of the validation parameters selectivity, linearity, accuracy and precision, analytical limits, and processed sample stability. Linearity was demonstrated over the whole calibration range from 5 to 1000 ng/ml with the use of a 1/x 2 weighting. Stability of the processed samples was verified for a time period of up to 25 h. Due to its high sensitivity, appropriate quantification and detection limits (LLoQ = 5 ng/ml, LoD = 0.95 ng/ml) for toxicological propofol analyses could be achieved. Applicability of the method to biological samples could be verified by analysis of a human serum sample collected after propofol-induced sedation. Graphical abstract A novel derivatization strategy using 1,2-dimethylimidazole-4-sulfonyl chloride (DMISC) was developed to improve the ionization and fragmentation efficacy of propofol for LC-ESI-MS/MS analysis.

Bite marks are rarely seen in forensic medical examinations. In most cases one can easily differentiate whether a bite mark is of human or animal origin due to the arched shape and crushed character of human bite marks. From a forensic point of view it is more interesting whether a dental imprint has been caused by an adult or a child, because children are often abused for a defensive statement. Two cases of bite injuries in children are presented. In the first case, a two-month-old infant was presented by pediatricians. The second case deals with a one-year-old boy who had allegedly been bitten several times by a two-year-old girl in a nursery school. A comparison could be performed between the bite mark impressions on the victim's skin (based on photographic documentation) and dental imprints of single teeth measuring the tooth width on the one hand and the intercanine distance on the other hand with the help of modeling wax plates. This procedure showed good correlations between the bite marks on the victim and the dental status of the biter. Especially for taking bite impressions on children, modeling wax plates are a simple, fast and cheap alternative to dental impression-taking.

Two rare causes of iatrogenic pericardial effusions are presented. In the first case, a 61-year-old woman who had undergone laparoscopic surgery for a diaphragmatic hernia was resuscitated without success the next day. As cause of death circulatory failure as a result of post-operative pulmonary embolism was reported. Autopsy results showed that the pericardium and the heart had been sewn to the diaphragm. The suture was torn from the tissue, which caused a hemorrhage into the pericardium and the chest cavity, so that death was diagnosed to be due to cardiac tamponade and hemothorax after an iatrogenic heart injury. In the second case, a 62-year-old man who had developed a massive incisional hernia after treatment of an abdominal gunshot wound underwent open herniotomy with mesh repair. Postoperatively, the man complained about increasing pain and shortness of breath. He was transferred to another hospital for further assessment, where a cardiac tamponade was diagnosed. Autopsy results showed that three of the plastic staples used to fix the mesh had perforated the diaphragm and the pericardium thus injuring the adjacent right ventricle with subsequent perforation and development of a hemopericardium.

The precision of death time estimation by vitreous potassium using 5 different formulas is compared on a random sample of 600 cases. Mean differences, standard deviations and 95% limits of confidence between real and extrapolated time since death were calculated. The best results were obtained using equations with "mean" slopes of about 0,17 or 0,19mmol/l per hour. Equations with a steeper or flatter slope reveal already great mean deviations between real and extrapolated time since death and do not allow a reliable death time estimation., (Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.)

Medicolegal death time estimation must estimate the time since death reliably. Reliability can only be provided empirically by statistical analysis of errors in field studies. Determining the time since death requires the calculation of measurable data along a time-dependent curve back to the starting point. Various methods are used to estimate the time since death. The current gold standard for death time estimation is a previously established nomogram method based on the two-exponential model of body cooling. Great experimental and practical achievements have been realized using this nomogram method. To reduce the margin of error of the nomogram method, a compound method was developed based on electrical and mechanical excitability of skeletal muscle, pharmacological excitability of the iris, rigor mortis, and postmortem lividity. Further increasing the accuracy of death time estimation involves the development of conditional probability distributions for death time estimation based on the compound method. Although many studies have evaluated chemical methods of death time estimation, such methods play a marginal role in daily forensic practice. However, increased precision of death time estimation has recently been achieved by considering various influencing factors (i.e., preexisting diseases, duration of terminal episode, and ambient temperature). Putrefactive changes may be used for death time estimation in water-immersed bodies. Furthermore, recently developed technologies, such as H magnetic resonance spectroscopy, can be used to quantitatively study decompositional changes. This review addresses the gold standard method of death time estimation in forensic practice and promising technological and scientific developments in the field.

In suicide cases involving an automobile, the car is mostly used as a tool/instrument for the suicide (intoxication by exhaust fumes, crash against solid objects such as a pylon). However, a car is uncommonly also used only as the location for a suicide. In the present case, a 70-year-old man had been found dead on the passenger seat of his car. According to witnesses, the car had not been parked there at the same time of the previous day. During that period, the weather had been dry, with temperatures ranging from -1⁰ C at night to +5⁰ C during the day. The man had shot himself with a small-caliber revolver (so-called bulldog-revolver) almost horizontally in the left temple (transtemporal shot through the frontal lobes). Obviously, the suicide had retained the capability to act at least for a short time, as the revolver was found under his left bottom. As morphological signs of prolonged agony some findings of hypothermia were observed (e. g. Wischnewski spots of the stomach).

Fatalities caused by cattle occur relatively often in rural areas and mostly show sharp and blunt injuries. In the presented case, a 61-year-old farmer was found dead in a cowshed together with an untied bull. Autopsy revealed severe hemorrhages and massive chest trauma with multiple bilateral rib fractures. The cause of death was severe chest trauma. In the presented case, death was not brought about by the bull's horns but by butts to the man's chest.

Fatal starvation is a rare cause of death in industrialized countries. However, it may have major medicolegal importance if death results from the deliberate withholding of food, especially from infants. In such cases, the task of the forensic pathologist and the medical examiner, respectively, is to clarify the cause of death and give an expert opinion on the degree and duration of starvation. Several classification systems have been developed to estimate protein-energy malnutrition in developing countries. Simpler classifications, such as the Gomez classification, use the weight expected for the respective age group as the standard. However, smaller infants will be lighter, and therefore the classification may not be accurate in this case. Following the Waterlow classification, the extent of stunted growth (referring to growth retardation in cases of chronic malnutrition) is calculated using the ratio of the measured body height to that expected for the age. Using such classification systems, grading of stunting and wasting can be achieved and may greatly help in the assessment of a given child's nutritional status in legal cases. The application of the Waterlow classification to the authors' case material and previously published cases in the literature is herein demonstrated. The Waterlow classification is not only of importance for grading the final stage of fatal starvation, but also for the chronological development of the nutritional status if anthropometrical data have been repeatedly recorded from the affected individual in vivo.

The sudden infant death syndrome (SIDS) is defined as unexpected death of a previously healthy infant with no cause of death being established. Comprehensive histological and immunohistochemical investigations are required subsequent to autopsy. In 18 cases of sudden infant death, histological (H&E, PAS and iron stain) and immunohistochemical investigations (CD68, CD45RO and LCA) were performed regarding a possible role of pancreatitis in SIDS and histomorphological alterations of the pancreatic islets, respectively. In all the investigated cases, no inflammatory changes of the infantile pancreatic tissue and no other significant histomorphological alterations of the pancreatic islets were found.

In a study on alcoholics, diabetics, cases of hypothermia, combinations of alcoholism, diabetes and hypothermia as well as 55 controls, ketone body measurements were performed in femoral vein blood, heart blood, vitreous humor, cerebrospinal fluid and urine. Histological investigations were carried out on the kidneys of the deceased. In addition to HE-staining, the cuts were stained with Sudan and PAS to allow differentiation between lipids and glycogens. The degree of stainability in the Sudan stains was correlated with the ketone body concentrations measured. In those cases in which elevated ketone body concentrations were measured, marked fat deposits in the renal tubular epithelial cells could be demonstrated with the Sudan staining method. The higher the stainability the higher the ketone body concentrations. The ketone body concentrations measured in the various body fluids correlated with the intensity of fat stainability.

Pancuronium(bromide) is used because of its relaxing effect on striated muscles and usually requires artificial respiration. A 52-year-old woman suffered from long-standing "generalized dystonia", which had become resistant to conventional therapy. Therefore, an anesthetist established a permanent medication scheme with pancuronium using a PCA pump. This pump had been controlled by the patient herself ensuring an acceptable quality of life with broad personal autonomy. Finally, the woman was found dead in her flat by a member of a home nursing service. The infusion hose showed a fixed knot and further blocking by a clamp. The autopsy findings were non-specific, except for the presence of opioid tablets in the colon. Toxicological analyses showed 72ng/ml pancuronium and 21 ng/ml oxycodone (therapeutic) in the femoral venous blood. The range of published pancuronium levels varies from approx. 80 to 2,000 ng/ml. Thus it had to be assumed that the pancuronium level was too low (72 ng/ml) so that symptoms of dystonia recurred. Based on extensive literature research, the described case can be qualified as unique. The therapy concept had been innovative, sufficient and effective for more than 10 years. It allowed the patient to enjoy a maximum of autonomy. Ultimately, death was due to the blocked pancuronium infusion. The relatively low pancuronium level had provoked the dystonia to return with generalized spasms also involving the respiratory muscles resulting in respiratory arrest. During the police investigations, two previous suicide attempts came to light.

This study evaluated the normal number of inflammatory cells in the heart in the first year of life using two methods to compare their ability to quantitate physiological myocardial infiltration. Eight endomyocardial samples from both ventricles were obtained at autopsy from 56 structurally normal hearts during the first year of life. In each sample the numbers of leucocytes, T-lymphocytes and macrophages were counted once in 20 randomly chosen high-power fields (400×) as well as in a 10mm(2) area of randomly chosen myocardial tissue (100×) by two independent investigators. Compared to the literature a greater representative proportion of myocardial tissue was analyzed. The results of the enumeration in mm(2) were converted into high-power-fields to compare both methods. The mean numbers and standard deviations for leucocytes, T-lymphocytes and macrophages were calculated. Both counting methods showed similar results with low inflammatory cell counts per single heart and staining. A greater understanding of the physiological myocardial infiltration by leucocytes, T-lymphocytes and macrophages is important for postmortem forensic cases, and for the interpretation of endomyocardial biopsies in infants., (Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.)

Fusobacteria belong to the normal population of the pharyngeal mucosa as well as the mucosa of the upper airways and the gastrointestinal tract. Infections are comparatively rare. The most common causative organism is Fusobacterium necrophorum. A well-known infection caused by this germ is Lemierre's syndrome. In the presented case, a 19-year-old man (123 kg body weight, 186 cm body length) was found dead in his bed in the morning after having complained of muscular fatigue and vomiting the previous day. Autopsy was carried out only two days after death. At that time, the body showed marked putrefaction with partial greenish discoloration and marbling of the skin although it had been stored in a refrigerator at +2 degrees C in the meantime. While the autopsy itself revealed no cause of death, microbiological examination of a smear from the left lower pulmonary lobe demonstrated Staphylococcus aureus and Fusobacterium necrophorum. Toxicological investigations produced negative results throughout. The cause of death was defined as sepsis caused by Fusobacterium necrophorum.

In most cases, bodily harm results from the use of sharp objects or blunt force. This paper deals with a 42-year-old pharmacist who was known to the police and the courts because of several previous convictions for bodily injury. The man had visited a pub just before it closed and was therefore not served any drinks. He got angry about this and returned to his pharmacy (within walking distance) to fetch three disposable syringes which he filled with phosphoric acid (85%). Through the open pub window, he splattered the acid from the syringes on two guests and the host, who were hit on the upper part of their bodies and the arms. All the victims developed dermal alterations such as focal erythema and small blisters (pemphigus-like efflorescences, as already described by Weyrich). At first, the pharmacist denied the use of phosphoric acid and claimed to have used a mixture of urine and water. Examinations of spots on the still unwashed clothes revealed very low pH-values (ca. 2.0; pH-Indicator-Stripes, Merck; Medi-Test, Machery & Nagel). Tests for substances typical of urine produced completely negative results. However, very high phosphate concentrations were found on the spots in question. Thus, the probability that the pharmacist had used phosphoric acid to commit the offence was very high. The pharmacist was sentenced to one year and two months' imprisonment for dangerous bodily harm according to Section 224 German Criminal Code. In accordance with the law, phosphoric acid was classified as "poison", for which application on the skin is sufficient.

The mitochondrial (mt) genome is present in many copies in human cells, and intra-individual variation in mtDNA sequences is known as heteroplasmy. Recent studies found that heteroplasmies are highly tissue-specific, site-specific, and allele-specific, however the functional implications have not been explored. This study investigates variation in mtDNA copy numbers (mtCN) in 12 different tissues obtained at autopsy from 152 individuals (ranging in age from 3 days to 96 years). Three different methods to estimate mtCN were compared: shotgun sequencing (in 4 tissues), capture-enriched sequencing (in 12 tissues) and droplet digital PCR (ddPCR, in 2 tissues). The highest precision in mtCN estimation was achieved using shotgun sequencing data. However, capture-enrichment data provide reliable estimates of relative (albeit not absolute) mtCNs. Comparisons of mtCN from different tissues of the same individual revealed that mtCNs in different tissues are, with few exceptions, uncorrelated. Hence, each tissue of an individual seems to regulate mtCN in a tissue-related rather than an individual-dependent manner. Skeletal muscle (SM) samples showed an age-related decrease in mtCN that was especially pronounced in males, while there was an age-related increase in mtCN for liver (LIV) samples. MtCN in SM samples was significantly negatively correlated with both the total number of heteroplasmic sites and with minor allele frequency (MAF) at two heteroplasmic sites, 408 and 16327. Heteroplasmies at both sites are highly specific for SM, accumulate with aging and are part of functional elements that regulate mtDNA replication. These data support the hypothesis that selection acting on these heteroplasmic sites is reducing mtCN in SM of older individuals.

On April 15th 2014, human skeletal remains were found during digging activities for constructing a new building at Bornheim-Uedorf (Rhein-Sieg-Kreis) near the river Rhine (about 20 meters) in a pit measuring 10 by 10 meters and having a depth of about 150 cm. The skeletal remains were preserved quite well considering the fact that they were located so near to the Rhine, although several skeletal parts were missing. The preserved skeletal remains comprised some skull fragments (including two ear bones of the right side), right scapula, both humeri, left ulna, left radius, left metacarpal bone 2, right metacarpal bones 1, 3 and 4, rib fragments, three thoracic vertebrae, all lumbar vertebrae, one sacral vertebra, pelvis fragments, left femur, proximal part of the left tibial diaphysis, right tibia and diaphysis of both fibulae. The anthropological analysis revealed that the skeletal remains belonged to a 20-to-30-year-old presumably male individual with a body height of about 163 to 173 cm (depending on the formula used for body height estimation). Evidence of intense physical activity and traumatological findings could not be detected on the preserved bones. Periosteal reactions on the bone surface caused by nonspecific bacterial infection were found on the right humerus close to the elbow and on both tibiae, especially the left one. Besides the skeletal remains, metal fragments were recovered--among them an arrowhead, which was typologically classified as an early medieval finding (6th to 7th century). Radiocarbon dating of a bone sample revealed an age of 1561 ± 19 a BP corresponding to a calibrated age of 436 to 540 AD (1 sigma). So, the archaeological classification of the recovered skeleton into the early medieval period was verified. Amongst the human remains, there was also a metatarsal bone of cattle with cut marks. The animal bone as well as the metal fragments indicated that the find was part of an early medieval burial with typical grave goods.

In the last few years, a growing number of cases have been reported in Germany in which vending machines have been blasted by criminals to get at the money. Thus, it was only a question of time for the first fatalities to occur as a consequence of such careless explosions. We report on the death of a 16-year-old boy who died after triggering an explosion by spraying a deodorant into the coin slot of a cigarette machine. Death was caused by severe craniocerebral trauma due to tertiary blast-related injuries when the front plate of the machine hit the victim's cerebral and facial skull.

Purpose: In industrialized countries, sudden infant death is the most common cause of death in young children. Although prone sleeping position is a well-known risk factor, hyperthermia might also be important. Pathognomonic findings of premortem hyperthermia do not exist. During stress, including thermal effects, heat-shock protein (HSP) expression increases. This study investigated hyperthermia as a contributing or pathogenic factor for sudden infant death syndrome (SIDS)., Methods: Immunohistochemical staining for HSP27 and HSP70 in the kidney, heart, and lung from 120 SIDS cases was examined., Results: HSP70 immunostaining was negative in kidney, heart, and lung tissues in all cases and in tissues from the control group. HSP27 staining was positive in the kidney from one case, and was positive in the lungs (respiratory epithelia in 27% of cases; vascular endothelia in 19% of cases) and was negative in the heart. In the control group HSP27 was positive in 8% of renal tubular tissues and in 29% of renal vascular endothelia. Staining for HSP27 in lung tissues was positive in respiratory epithelia in 8% of cases and for vascular endothelia in 29%, whereas tissues from the heart were positive in only 4%., Conclusion: The hypothesis of hyperthermia being a pathogenic factor for SIDS was not supported by immunohistochemical visualization of HSP70 or HSP27.

A very serious case of DFSA (drug facilitated sexual assault) is presented, in which a six-year-old girl died following sedation with γ-hydroxybutyric acid (GHB). She had been sexually abused by a relative. Samples of cardiac blood, bile, vitreous humour, liver, kidney, brain tissues and hair were analysed by a LC-MS/MS method. The following GHB concentrations were determined: cardiac blood: 150 mg/l; bile: 292mg/l; vitreous humour: 58mg/l; liver: 100 mg/kg; kidney: 124.5 mg/kg, brain: 110 mg/kg. Very high GHB levels were found in the proximal part of the hair sample (about 40.9 ng/mg). In distal segments of hair - up to 12 cm distant from the hair scalp - GHB concentrations were higher than the overall found endogenous range of 2-3 ng/mg. Police investigations revealed that the uncle had also administered GHB to the older half-sister. Therefore, a sample of her hair was analysed accordingly, but unremarkable results were obtained. Comparing our toxicological results with police investigations and the offender's statements it can be assumed that the 6-year-old girl had ingested GHB. By exclusion of other causes of death a lethal intoxication with GHB could be confirmed., (Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.)

Estimation of the time of immersion is a common problem in forensic medicine. In Germany since about 50 years a table for estimating the minimum time interval of immersion is used. This table was developed taking into account signs of progressive putrefaction and maceration and the monthly average water temperature. The reliability of this table was already checked some years ago. In the study presented, 33 further cases were evaluated. When the average monthly water temperatures used in the table are compared to the actual values, the temperatures in the river Rhine have risen during the last 40 years. Therefore, always the actual water temperature has to be measured and to be taken into consideration for the estimation of immersion time. Since there may be also fluctuations of the monthly water temperatures in cases of longer lasting immersion, a temperature profile has to be taken and the mean water temperature has to be calculated. Besides, the table of Reh should be adapted to the increased water temperatures.

When a firearm projectile hits a biological target a spray of biological material (e.g. blood and tissue) is ejected from the entrance wound and propelled back into the direction of the firearm. This phenomenon has been termed 'backspatter' and if backspattered biological material reaches the firearm on its backward trajectory it may persist on and be recovered from the firearm's inside surfaces. Molecular genetic analysis of backspatter generated by contact shots and shots from very short distances has already been demonstrated to critically contribute to victim identification and the reconstruction of firearm-related crimes. It is not known, however, up to what shooting distance can backspatter be found on firearms' inside surfaces and what influence the weapon's type and caliber has on backspatter attributes (e.g. reach, amount and distribution). Therefore, the present pilot study investigated the effect of serval combinations of shooting distances and types of firearms and ammunitions on the analyzability of co-extracted DNA and micro-RNA in samples of backspatter collected from interior and exterior surfaces of the firearms after experimental shootings employing standardized ballistic models. We demonstrate the limiting effect of shooting distance and the type of firearm on the yield of nucleic acids recovered from backspatter and the success rates of forensic DNA profiling and RNA based body-fluid and organ tissue identification in experimental shootings., (Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.)

Over one year, 7 autopsy cases with narrow diameters of the descending aorta were seen in the Institute of Forensic Medicine at the University of Bonn. The autopsy results were supplemented by chemical-toxicological, alcohol and histological examinations. In all cases middle-aged men (24-38 years) died suddenly and predominantly in a domestic environment. To some extent the inner aortic diameters ranged significantly below the expected values in relation to body surfaces or were lower than the 5th percentile related to men aged 45 years. In four cases, potentially lethal concentrations of alcohol or intoxications, e.g. with heroin, were noted. In one case, the blood sugar level was significantly elevated. In the other two cases, all further investigations were unremarkable.

In recent years, many synthetic cannabinoid (CB) receptor agonists have appeared on the market as constituents of herbal incense mixtures known as "spice". Contrary to the declared use, they are perorally consumed as a replacement for marijuana to get "high". In many cases, detailed information on the physicochemical and pharmacological properties of the synthetic compounds found in spice preparations is lacking. We have now evaluated a large series of heterocyclic compounds, 1,3-disubstituted indole and 2-azaindole derivatives known or assumed to be CB 1 receptor agonists, many of which have previously been identified in forensic samples. The mainly observed structural variations to circumvent restriction by law were bioisosteric exchanges of functional groups in known CB 1 agonists. We analyzed the structure-activity relationships of compounds at human CB 1 and CB 2 receptors based on affinities obtained in radioligand binding studies, and determined their efficacy in cAMP accumulation assays. Moreover, we investigated the activities of the compounds at the orphan G protein-coupled receptors GPR18 and GPR55 both of which are known to interact with cannabinoids. Most of the investigated compounds behaved as potent full agonists of CB 1 and CB 2 receptors with affinities in the low nanomolar to subnanomolar concentration range. Some compounds were moderately potent GPR55 antagonists, while none interacted with GPR18. Most derivatives were predicted to cross the blood-brain barrier as determined by bioinformatics tools. These data are useful for assessing synthetic cannabinoids and will be helpful for predicting pharmacological properties of novel compounds that appear on the illicit drug market.

Numerous theories on the pathomechanism of SIDS favor a multifactorial genesis whereby infections of the upper respiratory tract and the pulmonary tissue have a trigger function. The aim of the presented study was to prove the presence of inflammatory responses in pulmonary tissue in SIDS cases because various studies revealed contradictory results about the evidence of inflammatory pulmonary responses in SIDS cases. For this purpose, 10 cases with defined causes of death were examined in comparison to 75 cases of sudden infant death syndrome (SIDS) with regard to CD45RO, CD68 and LCA positive cells in pulmonary tissue. Furthermore, the cell adhesion molecules E-selectin, P-selectin and VCAM were investigated. In the study group, the immunohistochemical imaging of CD45RO and LCA positive cells showed mostly negative results. There were no significant differences between study and control group as to the number of cells expressing CD45R0, CD68 and LCA. The same was true for the adhesion molecules E-selectin, P-selectin and VCAM.