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Continuous development and rapid turnover of drug market of new psychoactive substances (NPS) make it difficult to obtain up-to-date analytical methods for efficient detection of intoxication cases with new substances: no analytical data and no previously published concentration values in biological samples are indeed available. In this context, we aim to report the first fatal case involving two newly emerging arylcyclohexylamine derivatives (a group of dissociative ketamine-based substances): 2-fluoro-deschloroketamine (2F-DCK) and 3-methoxyeticyclidine (3-MeO-PCE). A 42-year-old man was found dead at his home with three plastic bags of “research chemicals” powders near him. Comprehensive screenings of drugs and toxic compounds as well as more selective assays (performed using NMR, HS-GC-FID, LC-MS/MS and LC-HRMS methods) allowed (1) to identify the three unknown powders, 2F-DCK, 3-MeO-PCE, and 5-methoxy-N,N-dimethyltryptamine (5-MeO-DMT, a hallucinogenic tryptamine-related NPS), with purity above 95%, and (2) to determine peripheral blood (1780, 90, and 52 µg/L), urine (6.1, 6.3, and 2.2 mg/L), bile (12, 3.5, and 1.7 mg/L), and vitreous humour (1500, 66 and 155 µg/L) concentrations of 2F-DCK, 3-MeO-PCE and 5-MeO-DMT, respectively. In addition, toxicological results also revealed recent use of cannabis, cocaine, and amphetamine by the victim, and hair analysis draw pathway of addiction (including experiments with various other NPS) for several months before death. This fatality was considered as the consequence of respiratory depression in a poly-drug user due to a “cocktail effect” of concurrent intakes of 2F-DCK (mainly), 3-MeO-PCE, 5-MeO-DMT, amphetamine, and cocaine. In addition, this case report provides analytical data that could support subsequent toxicological result interpretation in forensic cases involving such arylcyclohexylamine derivatives.

The diagenesis of a bone in the postmortem period causes an identifiable deterioration in histology. This degradation is characterized by a collagenous alteration, which can be observed very early. In order to develop a method for determining a postmortem interval for medico-legal use, two ribs collected from six human bodies were studied prospectively over 2 years. Each bone was studied after staining with Sirius red to demonstrate the degradation of collagen as a function of time. This study demonstrated a time-based bone alteration characterized by the architectural degradation of the lamellar bone, without any microbial influence in this postmortem period. The staining was carried out by using Sirius red and correlated this alteration with a collagenic degradation by chemical hydrolysis owing to the affinity of this dye to the amino acids lysine, hydroxylysine, and arginine. Our work asserts that human bone samples that were studied in a controlled environment and analyzed for 24 months underwent a diagenetic trajectory whose main element was collagen hydrolysis.

•Bilirubinometer measurements on bruises are different from those on healthy skin.•Measurements on bruises showed increasing then decreasing values over time.•Peak values on bruises are mostly observed on the 4th or 5th day after injury.•No peak value was observed on bruises before the 3rd day after injury. The dating of bruises can be of crucial interest in clinical forensic medicine, but the macroscopic or photographic evaluation of a bruise has not been regarded as reliable. Several methods have thus been regularly evaluated for this purpose, with reflectance spectrometry yielding promising results. The objective of this study is to evaluate the value of a bilirubinometer as a tool for dating bruises on living victims. A clinical follow-up was carried out on patients presenting a bruise with a known onset. Using a bilirubinometer, we obtained daily measurements of the bruise (bili-bruise) and of healthy skin (bili-skin). Potential confounding factors were collected: age, sex, body mass index, trauma mechanism and Fitzpatrick skin phototype. We followed 20 patients for a total of 88 measurements of bruises. Bili-skin values showed significant differences according to skin phototype. Differences between the bili-bruise and bili-skin values (Δ-bili) followed an increase phase, peaking between 3 and 5 days, and then there was a decrease phase. No significant Δ-bili value differences were observed based on the suspected confounding factors. Our results are in favour of a peak Δ-bili value generally at day 4 or 5 post-trauma. Notably, decreasing values were not observed before day 3. Decreasing Δ-bili values would then indicate a bruise resulting from an injury formed at least 3 days before the first measurement. Complementary work confirming such data would enable improvement of the performance of bruise dating in forensic medicine.

The evaluation of the postmortem interval (PMI) is an important consideration in forensic medicine. The time between death and the discovery thereof is extremely important, not only in terms of the legal process, but also for the course of the investigation. At the time of the discovery of human bones, the evaluation of the PMI is extremely difficult. To date, there is no really effective tool for this evaluation. The objective of our study is to characterize the architectural modifications of human bones using micro-tomography (μCT) in the postmortem period. Ten bone samples were extracted from a single fresh human cranial vault (no freezing, no embalming) from a body donation. The samples were conserved in a controlled environment (a laboratory hood at 20 °C with 60% humidity) for 10 weeks. Every 2 weeks after the death and the sample extraction from the vault, the samples were analyzed with μCT (resolution 10 μm, Bruker HR1172 ®). The 3D analysis focused on general 3D data related to the trabeculae: the total volume (TV), the bone volume (BV), and the bone surface (BS), as well as specific trabeculae data; namely: the number of trabeculae (TbN), trabecular thickness (TbTh), the average distance between the trabeculae (TbSp), and the trabecular pattern factor (TbPf). Each sample was observed separately and compared to itself over time; no comparisons among samples were made. We used a linear mixed model for repeated measures (an unstructured covariance pattern model) and post hoc comparisons between each consecutive time using linear contrast with an SAS software package, release 9.3 (SAS Institute, Cary, NC). We showed a statistical difference for BS/BV (p = 0.046) over time, with a significant decrease during the first 2 weeks (p = 0.003; mean decrease, − 0.99, 95% CI [− 1.61; − 0.36]), while TbSp (p < 0.0001) showed a significant increase during the first 2 weeks (p < 0.0001; mean increase, 0.022, 95% CI, [− 0.016; − 0.028]).Our study highlighted architectural trabecular modifications on postmortem bones using the μCT. These modifications occurred at an early stage, during the first 2 weeks of conservation. Complementary work will need to be performed to allow for better understanding of the biological phenomena observed and to create databases of osseous taphonomy, starting with various individuals, mediums of conservation, types of bone, and with varying techniques for analysis.

Post-mortem interval (PMI) estimation is an important issue in forensic medicine, particularly for criminal purposes and legal limitation periods. The goal of the present study is to examine the evolution of the trabecular cranial vault bone after 4 weeks of conservation in a controlled environment with micro-tomography (μCT) analyses. Four bone samples were extracted from a fresh human cranial vault (a donation to science according to the French law) and conserved in an air-controlled environment. The samples were weighed and μCT scanned at a 10-μm resolution every week after death for a month. The μCT features were identical for every sample. Each set of data from the μCTs was reconstructed, registered, and analyzed in terms of the total volume, bone volume, bone surface, number of trabeculae, trabeculae thickness, and mean distance of the trabeculae. The samples were conserved in a glass box in 20 °C air with 60% humidity in a laboratory hood between each μCT acquisition. Descriptive statistics were determined. Each sample was observed and compared to itself over time. After 1 month of conservation, the mean bone volume (−1.9%), bone surface (−5.1%), and trabecular number (−12.35%) decreased, whereas the mean trabecular separation (+5.55%) and trabecular thickness (+12.7%) increased. Many variations (i.e., increases and decreases) were observed between the extraction of the sample and the end of the 4 weeks of conservation. The present observations may be explained by bone diagenesis. Previous observations have indicated that protein and lipid losses occur with bone weight and volume losses. These diagenesis effects may explain the trabecular modifications observed in the present work. We observed many bone variations with the μCT scans between the beginning and the end of the conservation that had no explanations. Additional studies, particularly studies involving statistics, need to be performed to confirm our observations and explain these results more clearly.

•Micro-tomography is useful for bone architecture analyses.•The precision of the device can be improved with registration.•Registration is not perfect, and residual imprecision can be observed.•A study about the error of precision of the device should be indicated before the use of μCT in forensic analysis. Micro-computed tomography (μCT) provides micrometric 3D images and has been used in forensic studies for anthropology pubis measurement or insect description for post mortem interval estimation. Studies have suggested using registration, a superimposing images method between a reference and a target. This technique avoids positioning bias and increase the precision of μCT. However, no clear study has reported the precision with μCT analysis before or after registration in a forensic field. One fresh post mortem sample of a human cranial vault was collected. Two successive μCT acquisitions (resolution 10μm) of it were performed without repositioning. The data from the second acquisition were copied and registered by two trained operators (operators 1 and 2). Operator 1 performed a second registration process after 1 week (operator 1 bis). The images were analysed. The bone volume (BV), bone surface (BS), number of trabeculae (TbN), trabecular thickness (TbTh) and mean trabecular distance (TbSp) were compared before and after registration. The mean (±SD), the coefficient of variation (%CV), and the precision error of the standard deviation absolute value and of the coefficient of variation between operators 1 and 2 (inter-subject variability) and between operator 1 and 1 bis (intra-subject variability) were calculated. We also collected the second phalanx of the 2nd, 3rd and 4th fingers on the hands of a second individual. Two successive scans (resolution 27μm) were performed without repositioning. A comparison (mean±SD of BV, BS, TbN, TbTh, TbSp) was made between the first and second scans with and without registration, and an ANOVA repeated measures procedure was performed. For the vault, we show that after 30 registrations for each operator (1, 2 and 1 bis), the mean and %CV were very close for each variable and between operators. For BV and BS, the difference in the mean value was approximately 0.01 (mm3 and mm2, respectively). The precision error was higher in the inter-subject registrations for each variable. The precision error magnitude for all variables was very low (<0.01) in absolute value and of %CV. For the fingers, the difference between the first and second scans may be approximately 50% without registration. We found that the second scan without registration is significantly different for BV (p=0.006), BS (p=0.007), TbN (p=0.019) and TbSp (p=0.002). Knowing the precision of the device (with and without registration) is important to ensure that the accuracy of the μCT results.

Post-mortem interval (PMI) estimation is an important issue in forensic medicine, particularly for criminal purposes and legal limitation periods. The goal of the present study is to examine the evolution of the trabecular cranial vault bone after 4 weeks of conservation in a controlled environment with micro-tomography ([mu]CT) analyses. Four bone samples were extracted from a fresh human cranial vault (a donation to science according to the French law) and conserved in an air-controlled environment. The samples were weighed and [mu]CT scanned at a 10-[mu]m resolution every week after death for a month. The [mu]CT features were identical for every sample. Each set of data from the [mu]CTs was reconstructed, registered, and analyzed in terms of the total volume, bone volume, bone surface, number of trabeculae, trabeculae thickness, and mean distance of the trabeculae. The samples were conserved in a glass box in 20 °C air with 60% humidity in a laboratory hood between each [mu]CT acquisition. Descriptive statistics were determined. Each sample was observed and compared to itself over time. After 1 month of conservation, the mean bone volume (-1.9%), bone surface (-5.1%), and trabecular number (-12.35%) decreased, whereas the mean trabecular separation (+5.55%) and trabecular thickness (+12.7%) increased. Many variations (i.e., increases and decreases) were observed between the extraction of the sample and the end of the 4 weeks of conservation. The present observations may be explained by bone diagenesis. Previous observations have indicated that protein and lipid losses occur with bone weight and volume losses. These diagenesis effects may explain the trabecular modifications observed in the present work. We observed many bone variations with the [mu]CT scans between the beginning and the end of the conservation that had no explanations. Additional studies, particularly studies involving statistics, need to be performed to confirm our observations and explain these results more clearly.

Bacterial translocation is a recently developed concept referring to the passage of microorganisms from the gastrointestinal tract to extraintestinal sites, such as the blood. This phenomenon occurs in healthy individuals, is increased in certain diseases, and is even more intense after death. Intestinal bacteria are major agents in postmortem changes, but the timing of bacterial translocation after death is not yet fully understood. This chapter provides information about the different aspects of the intestinal microbiota, the mechanisms involved in this translocation, and the postmortem changes affecting the thanatomicrobiome. The different factors influencing postmortem bacterial translocation are analyzed, as well as its relevance to the forensic practice, such as the identification of infectious agents at autopsy, the postmortem interval estimation, or the postmortem toxicological analysis.