Longitudinal visibility of MRI findings in living victims of strangulation.

Initial experiences with magnetic resonance imaging (MRI) of living strangulation victims demonstrated additional findings of internal injuries compared to the standard clinical forensic examination. However, existing studies on the use of MRI for this purpose mostly focused on the first 48 h after the incident. The aims of this study were (a) to evaluate the longitudinal visibility of MRI findings after violence against the neck by performing two MRI examinations within 12 days and a minimum of four days between both MRI scans and (b) to assess which MRI sequences were most helpful for the detection of injuries. Twenty strangulation victims participated in this study and underwent one (n = 8) or two (n = 12) MRI scans. The first MRI examination was conducted during the first five days, the second five to 12 days after the incident. Two blinded radiologists assessed the MRI data and looked for lesions in the structures of the neck. In total, 140 findings were reported in the 32 MRI examinations. Most of the findings were detected in the thyroid and the muscles of the neck. T2-weighted SPACE with fat suppression, T1-weighted TSE and T1-weighted MPRAGE were rated as the most helpful MRI sequences. Subjects who showed findings in the initial scan also demonstrated comparable results in the second scan, which was performed on average 8.4 days after the incident. Our results show that even up to 12 days after the incident, the criminal proceeding of strangulation cases may greatly profit from the information provided by an MRI examination of the neck in addition to the standard clinical forensic examination.

DEPArray™ single-cell technology: A validation study for forensic applications.

In forensics investigations, it is common to encounter biological mixtures consisting of homogeneous or heterogeneous components from multiple individuals and with different genetic contributions. One promising mixture deconvolution strategy is the DEPArray™ technology, which enables the separation of cell populations before genetic analysis. While technological advances are fundamental, their reliable validation is crucial for successful implementation and use for casework. Thus, this study aimed to 1) systematically validate the DEPArray™ system concerning specificity, sensitivity, repeatability, and contamination occurrences for blood, epithelial, and sperm cells, and 2) evaluate its potential for single-cell analysis in the field of forensic science. Our findings confirmed the effective identification of different cell types and the correct assignment of successfully genotyped single cells to their respective donor(s). Using the NGM Detect™ Amplification Kit, the average profile completeness for diploid cells was approximately 80%, with ∼ 290 RFUs. In contrast, haploid sperm analysis yielded an average completeness of 51% referring to the haploid reference profile, accompanied by mean peak heights of ∼ 176 RFUs. Although certain alleles of heterozygous loci in diploid cells showed strong imbalances, the overall peak balances yielded acceptable values above ≥ 60% with a mean value of 72% ± 0.21, a median of 77%, but with a maximum imbalance of 9% between heterozygous peaks. Locus dropouts were considered stochastic events, exhibiting variations among donors and cell types, with a notable failure incidence observed for TH01. Within the wet-lab experimentation with >500 single cells for the validation, profiling was performed using the consensus approach, where profiles were selected randomly from all data to better mirror real casework results. Nevertheless, complete profiles could be achieved with as few as three diploid cells, while the average success rate increased to 100% when using profiles of 6-10 cells. For sperms, however, a consensus profile with completeness >90% of the autosomal diploid genotype could be attained using ≥15 cells. In addition, the robustness of the consensus approach was evaluated in the absence of the respective reference profile without severe deterioration. Here, increased stutter peaks (≥ 15%) were found as the main artifact in single-cell profiles, while contamination and drop-ins were ascertained as rare events. Lastly, the technique's potential and limitations are discussed, and practical guidance is provided, particularly valuable for cold cases, multiple perpetrator rapes, and analyses of homogeneous mixed evidence.

Automated detection of fatal cerebral haemorrhage in postmortem CT data.

During the last years, the detection of different causes of death based on postmortem imaging findings became more and more relevant. Especially postmortem computed tomography (PMCT) as a non-invasive, relatively cheap, and fast technique is progressively used as an important imaging tool for supporting autopsies. Additionally, previous works showed that deep learning applications yielded robust results for in vivo medical imaging interpretation. In this work, we propose a pipeline to identify fatal cerebral haemorrhage on three-dimensional PMCT data. We retrospectively selected 81 PMCT cases from the database of our institute, whereby 36 cases suffered from a fatal cerebral haemorrhage as confirmed by autopsy. The remaining 45 cases were considered as neurologically healthy. Based on these datasets, six machine learning classifiers (k-nearest neighbour, Gaussian naive Bayes, logistic regression, decision tree, linear discriminant analysis, and support vector machine) were executed and two deep learning models, namely a convolutional neural network (CNN) and a densely connected convolutional network (DenseNet), were trained. For all algorithms, 80% of the data was randomly selected for training and 20% for validation purposes and a five-fold cross-validation was executed. The best-performing classification algorithm for fatal cerebral haemorrhage was the artificial neural network CNN, which resulted in an accuracy of 0.94 for all folds. In the future, artificial neural network algorithms may be applied by forensic pathologists as a helpful computer-assisted diagnostics tool supporting PMCT-based evaluation of cause of death.

Effect of vaporizing cannabis rich in cannabidiol on cannabinoid levels in blood and on driving ability - a randomized clinical trial.

The aim of this prospective, placebo-controlled, double-blind, randomized, cross-over study was to determine cannabinoid levels in blood and driving-related ability after single (S1) and repetitive (S2) vaporization of cannabis rich in cannabidiol (CBD) containing < 1% Δ9-etrahydrocannabinol (THC). Healthy adult volunteers (Nsingle = 27, Nrepetitive = 20) with experience in smoking vapor-inhaled two low-THC/CBD-rich cannabis products both with < 1% THC (product 1: 38 mg CBD, 1.8 mg THC; product 2: 39 mg CBD, 0.6 mg THC) and placebo. Main outcomes were THC- and CBD-levels in whole blood and overall assessment of driving-related ability by computerized tests. Among 74 participants included, 27 (mean age ± SD, 28.9 ± 12.5 years) completed S1, and 20 (25.2 ± 4.0) completed S2. Peak concentrations and duration of detectability depended on the THC-content of the product. After single consumption THC dropped below 1.5 µg/L after 1.5 h, but was detected in some participants up to 5 h. Pairwise comparison of driving-related ability revealed no significant differences between low-THC/CBD-rich products (P1, P2) and placebo. Detection of THC after consumption of low-THC/CBD-rich cannabis might have legal consequences for drivers. Regarding overall driving-related ability, no significant differences were observed between the interventional products. This trial was registered with the German Clinical Trials Register (DRKS00018836) on 25.10.2019 and with the Coordination Office for Human Research (kofam) which is operated by the Federal Office of Public Health (FOPH) (SNCTP000003294).

A systematic approach to improve downstream single-cell analysis for the DEPArray™ technology.

Most commercially available STR amplification kits have never been fully validated for low template DNA analysis, highlighting the need for testing different PCR kits and conditions for improving single-cell profiling. Here, current strategies rely mainly on adjusting PCR cycle number and analytical threshold settings, with a strong preference for using 30 amplification cycles and thresholds at 30-150 RFU for allele detection. This study aimed to (1) determine appropriate conditions for obtaining informative profiles utilizing a dilution series, and (2) test the outcome on single cells using the DEPArray™ technology. Four routinely applied forensic STR kits were compared by using three different amplification volumes and DNA dilutions down to 3.0 pg, while two well-performing kits were used for single/pooled leucocyte and sperm cell genotyping. Besides reduced costs, the results demonstrate that a 50%-75% PCR volume reduction was beneficial for peak height evaluation. However, this was counteracted by an increased artifact generation in diluted DNA volumes. Regarding profile completeness, the advantage of volume reduction was only prominent in samples processed with Fusion 6C. For single and pooled cells, ESIFast and NGMDetect provided a solid basis for consensus profiling regarding locus failure, although locus dropouts were generally observed as stochastic events. Amplification volume of 12.5 µL was confirmed as appropriate in terms of peak heights and stutter frequencies, with increased stutter peaks being the main artifact in single-cell profiles. Limitations associated with these analyses are discussed, providing a solid foundation for further studies on low template DNA.

Temperature correction of post mortem quantitative magnetic resonance imaging using real-time forehead temperature acquisitions.

Performing magnetic resonance imaging (MRI) of deceased is challenging due to altered body temperatures compared to in vivo temperatures and, hence, requires a temperature correction. This study investigates the possibility to correct brain MRI parameters real-time and non invasively based on the forehead temperature. 17 post mortem cases were included and their forehead temperatures were measured continuously during the in situ brain MRI protocol consisting of a diffusion tensor imaging, multi-contrast spin echo, multi-echo gradient echo and inversion recovery spin echo sequence. Linear models were fitted to the quantitative MRI parameters in a forensically interesting temperature range for white matter, cerebral cortex and deep gray matter, separately, and the influence of the forehead temperature on the MRI parameters was determined. A statistically significant temperature sensitivity was found for T2 and mean diffusivity in white matter, for T1 in cerebral cortex, as well as for T1 and mean diffusivity in deep gray matter. Linear models were computed to temperature correct these MRI parameters in in situ post mortem scans to allow their comparison regardless of temperature. The here presented real-time and non invasive temperature correction method for the brain presents a crucial precondition for quantitative in situ post mortem MRI.

Investigation of post mortem brain, rectal and forehead temperature relations.

It is well known that magnetic resonance (MR) imaging is temperature sensitive, which is highly relevant for post mortem examinations. Therefore, the determination of the exact temperature of the investigated body site, e.g. the brain, is crucial. However, direct temperature measurements are invasive and inconvenient. Thus, in view of post mortem MR imaging of the brain, this study aims at investigating the relation between the brain and the forehead temperature for modelling the brain temperature based on the non-invasive forehead temperature. In addition, the brain temperature will be compared to the rectal temperature. Brain temperature profiles measured in the longitudinal fissure between the brain hemispheres, as well as rectal and forehead temperature profiles of 16 deceased were acquired continuously. Linear mixed, linear, quadratic and cubic models were fitted to the relation between the longitudinal fissure and the forehead and between the longitudinal fissure and the rectal temperature, respectively. Highest adjusted R2 values were found between the longitudinal fissure and the forehead temperature, as well as between the longitudinal fissure and the rectal temperature using a linear mixed model including the sex, environmental temperature and humidity as fixed effects. The results indicate that the forehead, as well as the rectal temperature, can be used to model the brain temperature measured in the longitudinal fissure. Comparable fit results were observed for the longitudinal fissure-forehead temperature relation and for the longitudinal fissure-rectal temperature relation. Combined with the fact that the forehead temperature overcomes the problem of measurement invasiveness, the results suggest using the forehead temperature for modelling the brain temperature in the longitudinal fissure.

Fully automated radiologic identification focusing on the sternal bone.

A crucial task in forensic investigations is the identification of unknown deceased. In general, secure identification methods rely on a comparison of ante mortem (AM) with post mortem (PM) data. However, available morphologic approaches are often dependent on the expertise and experience of the examiner, and often lack standardisation and statistical evidence. The objective of this study was therefore to overcome the current challenges via developing a fully automated radiologic identification (autoRADid) method based on the sternal bone. An anonymised AM data set consisting of 91 chest computed tomography (CT) scans, as well as an anonymised PM data set of 42 chest CT scans were included in this work. Out of the 91 available AM CT data sets, 42 AM scans corresponded to the 42 PM CT scans. For the fully automated identification analysis, a custom-made python pipeline was developed, which automatically registers AM data to the PM data in question using a two-step registration method. To evaluate the registration procedure and subsequent identification success, the image similarity measures Jaccard Coefficient, Dice Coefficient, and Mutual Information were computed. The highest value for each metric was retrieved in order to analyse the correspondence between AM and PM data. For all three similarity measures, 38 out of the 42 cases were matched correctly. This corresponds to an accuracy of 91.2%. The four unsuccessful cases incorporated surgical interventions taking place between the AM and the PM CT acquisition or poor CT scan quality preventing robust registration results. To conclude, the presented autoRADid method seems to be a promising fully automated tool for a reliable and facile identification of unknown deceased. A final pipeline combining all three similarity measures is open source and publicly available for efficient future identifications of unknown deceased.

The DNA-Buster: The evaluation of an alternative DNA recovery approach.

Touch DNA recovery techniques can have limitations, as their effectiveness depends on the substrate on which the DNA of a person of interest can be found. In this study, an in-house dry-vacuuming device, the DNA-Buster, was compared to traditional methods for its DNA recovery performance from items typically examined in forensic casework. The aim was to evaluate whether this dry-vacuuming approach can recover DNA efficiently, potentially complementing the well-established recovery strategies. For this, the performances of swabbing, taping, wet- (M-Vac®) and dry-vacuuming (DNA-Buster) were investigated quantitatively and qualitatively for touch DNA deposited on carpet, cotton sweater, stone, tile and wood. For the sweater, both vacuuming methods outperformed the other collection tools quantitatively. While the highest DNA amounts for the carpet were yielded by swabbing and taping, dry-vacuuming was equally good in reaching full DNA profiles, whereas less complete profiles were observed for the M-Vac®. For stone and tile, swabbing was optimal, whereas dry-vacuuming clearly underperformed for these substrates. Taping was the best recovery method for wood. Despite applying single donor DNA after thoroughly cleaning the items, undesired DNA mixtures were detected for all recovery techniques and all substrates. The overall research findings show first that the novel dry-vacuuming method is suited for DNA recovery from textiles. Secondly, they indicate that more attention should be paid to the substrate-collection dependency to ensure best practices in recovering genetic material in a precise, confident and targeted manner from the variety of forensic casework material.

Recommendations for the successful identification of altered human remains using standard and emerging technologies: Results of a systematic approach.

Successful DNA-based identification of altered human remains relies on the condition of the corpses and varies between tissue types. Therefore, the aim of this prospective multicenter study was to generate evidence-based recommendations for the successful identification of altered remains. For this, 19 commonly used soft and hard tissues from 102 altered human bodies were investigated. The corpses' condition was categorized into three anatomical regions using a practical scoring system. Besides other data, DNA yields, degradation indices, and short tandem repeat (STR) profile completeness were determined in 949 tissue samples. Additionally, varying degrees of alteration and tissue-specific differences were evaluated using the Next Generation Sequencing (NGS) platform MiSeq FGx™. Selected challenging samples were sequenced in parallel with the Ion S5™ platform to assess platform-specific performances in the prediction of the deceased's phenotype and the biogeographic ancestry. Differences between tissue types and DNA extraction methods were found, revealing, for example, the lowest degradation for vertebral disc samples from corpses with initiating, advanced and high degrees of decomposition. With respect to STR profile completeness, blood samples outperformed all other tissues including even profoundly degraded corpses. NGS results revealed higher profile completeness compared to standard capillary electrophoresis (CE) genotyping. Per sample, material and degradation degree, a probability for its genotyping success, including the "extended" European Standard Set (eESS) loci, was provided for the forensic community. Based on the observations, recommendations for the alteration-specific optimal tissue types were made to improve the first-attempt identification success of altered human remains for forensic casework.

Technical note: Quantitative optimization of the FLAIR sequence in post mortem magnetic resonance imaging.

The fluid attenuated inversion recovery (FLAIR) magnetic resonance imaging (MRI) sequence aims at suppressing the signal of the cerebrospinal fluid (CSF) by acquiring images at the time point at which the longitudinal magnetization and therefore the signal of CSF is zero. This time point is also called the null point inversion time (TInull). However, the FLAIR sequence is impaired by the temperature dependency of TInull in post mortem MRI due to the lower body temperature of the deceased subject. Therefore, the temperature correction of TInull is crucial for correctly suppressing the CSF signal in post mortem FLAIR imaging. Thus, the goal of this study was to determine the temperature effect on post mortem TInull for achieving a robust suppression of the CSF signal in in situ post mortem MRI using the FLAIR sequence. For this purpose, nine deceased subjects underwent an in situ MRI brain examination on a 3 T MRI scanner. TInull of CSF was determined quantitatively based on different FLAIR acquisitions using varying inversion times. The brain and rectal temperatures were determined prior to the MRI scan. A significant positive linear relation was found between TInull of CSF and the brain temperature, as well as between TInull of CSF and the rectal temperature. The found linear relations between TInull and both brain and rectal temperatures allow correcting TInull for varying temperatures of the deceased. This in turn enables an optimal suppression of the CSF signal in future post mortem FLAIR MRI acquisitions.

Post mortem evaluation of brain edema using quantitative MRI.

The post mortem assessment of brain edema is routinely performed during autopsy by forensic pathologists who evaluate the macroscopically visible signs. In this study, the suitability of magnetic resonance imaging (MRI) for a differentiation between edematous and nonedematous brains was examined as an objective, noninvasive and quantitative rating method. In this study, 22 deceased underwent post mortem in situ MRI prior to regular autopsy. Acquired MRI sequences allowed the computation of the quantitative MRI parameters T1, T2, T2*, fractional anisotropy (FA) and mean diffusivity for the cortex, white matter and deep gray matter separately. Beside the autopsy results, which represent the gold standard for rating brain edema, also the normalized cerebral weight (NCW) was determined by dividing the brain weight by the intracranial volume as developed by Bauer et al. [10]. For further examination of the relation of MRI parameters with the NCW, linear regression models were calculated. The results revealed highly significant correlations of the MRI parameters T2 and FA with the NCW in the cortex. These combinations additionally showed the best fitting results of the linear regression models. In conclusion, quantitative MRI is suitable for differentiating edematous from nonedematous brains by calculating T2 and FA in the cortex. A post mortem in situ MRI scan of the brain can, besides delivering morphological information, add relevant and objective information on the edema status of the brain prior to autopsy or when no autopsy is ordered.

Determination of time of death by blinded post-mortem interrogation of cardiac implantable electrical devices.

Postmortal interrogation of cardiac implantable electrical devices (CIED) may contribute to the determination of time of death in forensic medicine. Recent studies aimed to improve estimation of time of death by combining findings from autopsy, CIED interrogation and patients´ medical history. CIED from deceased undergoing forensic autopsy were included, if time of death remained unclear after forensic assessment. CIED explanted from deceased with known time of death were analysed as a control cohort. CIED were sent to our device interrogation lab and underwent analysis blinded for autopsy findings, medical history and police reports. The accuracy of time of death determination and the accuracy of time of death in the control cohort served as primary outcome. A total of 87 CIED were analysed. The determination of time of death was possible in 54 CIED (62%, CI 52-72%). The accuracy of the estimated time of death was 92.3% in the control cohort. Certain CIED type and manufacturers were associated with more successful determination. Blinded postmortal analysis enables a valid determination of the time of death in the majority of CIED. Analysis of explanted CIED in a cardiological core lab is feasible and should be implemented in forensic medicine.

Beyond Δ9-tetrahydrocannabinol and cannabidiol: chemical differentiation of cannabis varieties applying targeted and untargeted analysis.

Cannabis sativa (C. sativa) is commonly chemically classified based on its Δ9-tetrahydrocannabinol (THC) and cannabidiol (CBD) content ratios. However, the plant contains nearly 150 additional cannabinoids, referred to as minor cannabinoids. Minor cannabinoids are gaining interest for improved plant and product characterization, e.g., for medical use, and bioanalytical questions in the medico-legal field. This study describes the development and validation of an analytical method for the elucidation of minor cannabinoid fingerprints, employing liquid chromatography coupled to high-resolution mass spectrometry. The method was used to characterize inflorescences from 18 different varieties of C. sativa, which were cultivated under the same standardized conditions. Complementing the targeted detection of 15 cannabinoids, untargeted metabolomics employing in silico assisted data analysis was used to detect additional plant ingredients with focus on cannabinoids. Principal component analysis (PCA) was used to evaluate differences between varieties. The overall purpose of this study was to examine the ability of targeted and non-targeted metabolomics using the mentioned techniques to distinguish cannabis varieties from each other by their minor cannabinoid fingerprint. Quantitative determination of targeted cannabinoids already gave valuable information on cannabinoid fingerprints as well as inter- and intra-variety variability of cannabinoid contents. The untargeted workflow led to the detection of 19 additional compounds. PCA of the targeted and untargeted datasets revealed further subgroups extending commonly applied phenotype classification systems of cannabis. This study presents an analytical method for the comprehensive characterization of C. sativa varieties.

Validation and beyond: Next generation sequencing of forensic casework samples including challenging tissue samples from altered human corpses using the MiSeq FGx system.

The proceeding developments in next generation sequencing (NGS) technologies enable increasing discrimination power for short tandem repeat (STR) analyses and provide new possibilities for human identification. Therefore, the growing relevance and demand in forensic casework display the need for reliable validation studies and experiences with challenging DNA samples. The presented validation of the MiSeq FGx system and the ForenSeq™ DNA Signature Prep Kit (1) investigated sensitivity, repeatability, reproducibility, concordance, pooling variations, DNA extraction method variances, DNA mixtures, degraded, and casework samples and (2) optimized the sequencing workflow for challenging samples from human corpses by testing additional PCR purification, pooling adjustments, and adapter volume reductions. Overall results indicate the system's reliability in concordance to traditional capillary electrophoresis (CE)-based genotyping and reproducibility of sequencing data. Genotyping success rates of 100% were obtained down to 62.5 pg DNA input concentrations. Autosomal STR (aSTR) profiles of artificially degraded samples revealed significantly lower numbers of locus and allelic dropouts than CE. However, it was observed that the system still exposed drawbacks when sequencing highly degraded and inhibited samples from human remains. Due to the lack of studies evaluating the sequencing success of samples from decomposed or skeletonised corpses, the presented optimisation studies provide valuable recommendations such as an additional PCR purification, an increase in library pooling volumes, and a reduction of adapter volumes for samples with concentrations ≥31.2 pg. Thus, this research highlights the importance of all-encompassing validation studies for implementing novel technologies in forensic casework and presents recommendations for challenging samples.

Quantitative determination of five cannabinoids in blood and urine by gas chromatography tandem mass spectrometry applying automated on-line solid phase extraction.

Cannabis is the most frequently consumed illegal substance worldwide. More recently, an increasing number of legal cannabis products low in psychoactive Δ9 -tetrahydrocannabinol (THC) but high in non-intoxicating cannabidiol (CBD) are being more widely consumed. While the detection and quantification of THC and its metabolites in biological matrices is an important forensic-toxicological task, additional detection of CBD is also important, for example, when examining the plausibility of consumer's statements. This report describes the method validation for the quantitative determination of THC and its two major metabolites, 11-hydroxy-THC (OH-THC) and 11-nor-9-carboxy-THC (THC-COOH), as well as CBD and cannabinol (CBN) in whole blood and urine. The method employs automated on-line solid phase extraction coupled to gas chromatography tandem mass spectrometry (GC-MS/MS). The method was fully validated according to guidelines of the Swiss Society of Legal Medicine (SGRM) and the Society of Toxicological and Forensic Chemistry (GTFCh). The method fulfilled the validation criteria regarding analytical limits, accuracy and precision, extraction efficacy, and sample stability. The limits of detection (LODs) in whole blood and urine were 0.15 ng/mL for THC, OH-THC and CBD, 0.1 ng/mL for CBN, and 1.0 ng/mL for THC-COOH. The limits of quantification (LOQ) in whole blood and urine were 0.3 ng/mL for THC, OH-THC and CBD, 0.2 ng/mL for CBN, and 3.0 ng/mL for THC-COOH. The fully validated and automated method allows sensitive and robust measurement of cannabinoids in whole blood and urine. Detection of CBD provides additional information regarding consumed products.

Adulteration of low-delta-9-tetrahydrocannabinol products with synthetic cannabinoids: Results from drug checking services.

Since late 2019, low-delta-9-tetrahydrocannabinol (THC) preparations adulterated with synthetic cannabinoids (SCs) have been frequently observed in Switzerland. The unawareness of users concerning the presence of SCs and the typically higher potency and toxicity of SCs, when compared with THC, can result in increased health risks. In Switzerland, low-THC (<1%) cannabis products, except hashish, are legal. These products can act as carrier materials for SCs. In this study, cannabis samples and user self-reports received through three drug checking services were collected and analysed, to gain deeper insight into this new phenomenon. Samples were collected from January 2020 to July 2021. Liquid chromatography coupled with high-resolution mass spectrometry was used for the qualitative screening and semi-quantification of SCs, while gas chromatography with flame ionization detector was applied for the quantification of THC and cannabidiol levels. Reported adverse effects were compared between users who consumed adulterated (SC-group) and non-adulterated (THC-group) products. Of a total 94 samples, 50% contained up to three different SCs. MDMB-4en-PINACA was most often detected. All adulterated cannabis flowers contained ≤1% THC. Adulterated hashish also typically presented low THC-levels (median: 0.8%). The SC-group was associated with higher numbers of adverse events (p = 0.041). Furthermore, psychologic (p = 0.0007) and cardiologic (p = 0.020) adverse effects were more profound in the SC-group than in the THC-group. Drug checking services enabled the timely detection and monitoring of new and potentially dangerous trends. Furthermore, due to user-reports, additional valuable information was gained on adverse events associated with the consumption of novel SCs.

Post mortem brain temperature and its influence on quantitative MRI of the brain.

OBJECTIVE: MRI temperature sensitivity presents a major issue in in situ post mortem MRI (PMMRI), as the tissue temperatures differ from living persons due to passive cooling of the deceased. This study aims at computing brain temperature effects on the MRI parameters to correct for temperature in PMMRI, laying the foundation for future projects on post mortem validation of in vivo MRI techniques. MATERIALS AND METHODS: Brain MRI parameters were assessed in vivo and in situ post mortem using a 3 T MRI scanner. Post mortem brain temperature was measured in situ transethmoidally. The temperature effect was computed by fitting a linear model to the MRI parameters and the corresponding brain temperature. RESULTS: Linear positive temperature correlations were observed for T1, T2* and mean diffusivity in all tissue types. A significant negative correlation was observed for T2 in white matter. Fractional anisotropy revealed significant correlations in all gray matter regions except for the thalamus. DISCUSSION: The linear models will allow to correct for temperature in post mortem MRI. Comparing in vivo to post mortem conditions, the mean diffusivity, in contrast to T1 and T2, revealed additional effects besides temperature, such as cessation of perfusion and active diffusion.

Which tissue to take? A retrospective study of the identification success of altered human remains.

In forensic medicine, deceased are usually identified by comparing ante- and post-mortem dental or radiological features. However, in severe putrefaction, burning or absent reference data, the remaining tool for identifying human remains is DNA genotyping. But even a DNA-based identification can be challenging when confronted with a high post-mortem interval or heat impacts because it can lead to undesirable degradation of the DNA that varies among tissue types. This retrospective study investigated the identification success in 402 altered human corpses over seven years by comparing the examined tissue types from decomposed, skeletonised and burnt corpses as well as bodies found in water. For each tissue, the STR genotyping results and the number of additional or parallel genetic analyses were evaluated. By comparing the amplification success in samples from altered and unaltered remains, condition-based and tissue-specific differences were observed. With a mean number of 1.6 additional amplifications in cases with well-preserved corpses and 4.5 in altered corpses, the results showed significantly more DNA analyses for altered remains. In 83% of the cases, extra amplifications were performed to identify the corpse. The tissue-specific differences revealed an uncertainty in choosing suitable material from altered corpses for a successful DNA profile. Especially for bone and muscle samples, the genotyping success was the most unpredictable. Furthermore, comparing the retrospective outcome with other research findings, a remarkable variety of recommendations for the "best tissue choice" exists in the forensic community. Thus, our survey highlights the advantages of a broader and systematic approach on hard and soft tissues for successful DNA-based identification of altered human remains at first attempt.

Phase I In Vitro Metabolic Profiling of the Synthetic Cannabinoid Receptor Agonists CUMYL-THPINACA and ADAMANTYL-THPINACA.

Synthetic cannabinoid receptor agonists (SCRAs) remain popular drugs of abuse. As many SCRAs are known to be mostly metabolized, in vitro phase I metabolic profiling was conducted of the two indazole-3-carboxamide SCRAs: CUMYL-THPINACA and ADAMANTYL-THPINACA. Both compounds were incubated using pooled human liver microsomes. The sample clean-up consisted of solid phase extraction, followed by analysis using liquid chromatography coupled to a high resolution mass spectrometer. In silico-assisted metabolite identification and structure elucidation with the data-mining software Compound Discoverer was applied. Overall, 28 metabolites were detected for CUMYL-THPINACA and 13 metabolites for ADAMATYL-THPINACA. Various mono-, di-, and tri-hydroxylated metabolites were detected. For each SCRA, an abundant and characteristic di-hydroxylated metabolite was identified as a possible in vivo biomarker for screening methods. Metabolizing cytochrome P450 isoenzymes were investigated via incubation of relevant recombinant liver enzymes. The involvement of mainly CYP3A4 and CYP3A5 in the metabolism of both substances were noted, and for CUMYL-THPINACA the additional involvement (to a lesser extent) of CYP2C8, CYP2C9, and CYP2C19 was observed. The results suggest that ADAMANTYL-THPINACA might be more prone to metabolic drug-drug interactions than CUMYL-THPINACA, when co-administrated with strong CYP3A4 inhibitors.