In the case of extensively putrefied bodies, the analysis of entomological samples may support and complement the toxicological results obtained with other alternative matrices.

Entomotoxicological analysis is not part of routine toxicological analysis. This work aims to present two cases to illustrate the potential of entomological samples as complementary matrices to identify substances in cases of advanced putrefaction. (Case#1) A woman wasexhumed after 14 months to ascertain the exact cause of death. She died after six weeks of hospitalization because of intestinal ischemia followed by multiorgan failure. (Case#2) The corpse of a woman, known to have a psychiatric disorder, was discovered in her apartment. The state of decomposition of the body was consistent with a post-mortem period of several weeks (approximately 6 weeks). Toxicological investigations were performed in the biological and entomological samples of case#1 (hair, adipocere, brain, and pupae) and of case#2 (hair, bone, flies, and pupae) using liquid chromatography with high-resolution mass spectrometry and tandem mass spectrometry detection methods. In case#1, several drugs and metabolites were detected. In particular, the pupae analyses allowed the objectification of morphine administration, whereas morphine was only found in adipocere, but not in hair nor in brain. In case#2, the pupae analyses allowed the detection of three metabolites of quetiapine, and the flies analyses allowed the detection of valpromide, which was only detected in hair. In conclusion, the pupae and flies analyses in these two cases complemented the results obtained in the other alternative biological samples, which may guide hypotheses about the possible causes of death. Nevertheless, additional data and case reports would be of benefit to assess the value of entomotoxicology in routine forensic investigations.

Metabolite elucidation of 2-fluoro-deschloroketamine (2F-DCK) using molecular networking across three complementary in vitro and in vivo models.

This work first aims to investigate metabolites of 2-fluoro-deschloroketamine (2F-DCK), a new arylcyclohexylamine derivatives (a group of dissociative ketamine-based substances) using two in vitro experimental approaches, and to compare obtained results by means of molecular networking. Metabolites of 2F-DCK were investigated using both human liver microsomes (HLMs) and hepatic (HepaRG) cell line incubates using molecular networking approach: 2F-DCK pure substance was incubated with HLMs for up to 1 h at two concentrations (100 and 500 µM) and with HepaRG cells for two time periods (8 and 24 h) at one concentration (20 µM). In vitro obtained results were subsequently applied to a 2F-DCK-related fatality case. In vitro-produced metabolites were investigated using high-resolution accurate mass spectrometry using Orbitrap mass analyzer technology. Thirteen metabolites were in vitro produced and several metabolic pathways can be postulated. Seven additional metabolites were found in post-mortem samples (bile and urine) of the case, comprising three Phase II metabolites, which appear to be minor in vivo metabolites. HLMs and HepaRG cell models appear to be complementary and obtained data allowed the identification of several specific 2F-DCK metabolites in biological samples. In practical terms, observed metabolic ratios suggested that nor-2F-DCK (208.1137 m/z) and a hydrogenated metabolite (224.1443 m/z) could be proposed as reliable metabolites to be recorded in HRMS libraries in order to improve detection of 2F-DCK use.

Fatal intoxication related to two new arylcyclohexylamine derivatives (2F-DCK and 3-MeO-PCE).

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.

Medico-legal considerations for Morel-Lavallée lesions.

The Morel-Lavallée lesion is an infrequent traumatic lesion that the forensic physician may need to evaluate during examination of a victim. Using a review of the literature and an illustrative case, the objective of this paper is to discuss the medico-legal issues (accountability, functional recovery and healing times, and aftermath) associated with Morel-Lavallée lesions. We describe the case of a 24-year-old motorcycle driver who was hit by a car. Clinically, the right lumbar region had a large 16 × 15 cm ecchymosis with subcutaneous fluid swelling found by palpation. A body scan revealed a Morel-Lavallée lesion among other lesions. On day 7, its thickness was decreased by half. Morel-Lavallée lesions are specifically induced by shearing force with moderate-to-high kinetics in an anatomical area with an underlying fascia. The associated functional disability is globally moderate, but a large lesion or recurrences can extend the time needed to recover. Apart from some exceptions, care without any delay in diagnosis and treatment allows evolution of the lesion without functional consequences, but these lesions are frequently unnoticed in the initial phase.

Determination of the age of bruises using a bilirubinometer.

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.

Fatal Overdose of Gamma-hydroxybutyrate Acid After Ingestion of 1,4-Butanediol.

We report a case of fatal intoxication from 1,4-butanediol (1,4-BD), which was ingested by a young and "naïve" gamma-hydroxybutyrate (GHB) consumer during a party with the co-ingestion of alcohol, cannabis, and methylene-dioxy-methamphetamine. The following drug concentrations were found using gas chromatography coupled with mass spectrometry on autopsy samples and on a cup and a glass found at the scene: 20,350 mg/L (bottle) for 1,4-BD; 1020 mg/L (femoral blood), 3380 mg/L (cardiac blood), 47,280 mg/L (gastric content), and 570 mg/L (vitreous humor) for GHB. The concentration of GHB is difficult to interpret in forensic cases due to the possibility of an endogenous production of GHB. The variable tolerance of the user may also modify the peri- and postmortem GHB concentrations. This case underscores the need to have many different sources of toxicology samples analyzed to avoid the hypothesis of endogenous production of GHB.

Is bone analysis with µCT useful for short postmortem interval estimation?

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.

Characterization of bone diagenesis by histology in forensic contexts: a human taphonomic study.

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.

Domestic Predation of an Elder: A Fatal Dog Attack Case.

We present the case of a 91-year-old woman lived alone at her home with two domestic dogs, that is,, a Labrador Retriever and a Staffordshire Bull Terrier and found dead. The investigation of the scene revealed that the Bull Terrier's jawbone and chest were covered with blood. The autopsy revealed multiple, histologically confirmed, life-threatening skin and bone lacerations without scavenging marks. The punctures and tearing of each of the wounds on the skin were compatible with bites. A left humeral fracture and multiple fractures of the right facial bones were observed. The death was attributed to external hemorrhages due to several dog bites. A veterinary physical and behavioral examination indicated that the Bull Terrier was involved in the attack. A domestic predation hypothesis was deemed here most likely due to the presence of food supplies at the scene, the dog's previous history of attack, and the breed of the dog.

The precision of micro-tomography in bone taphonomic experiments and the importance of registration.

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.

Technical note: early post-mortem changes of human bone in taphonomy with µCT.

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.

Cyanide Suicide After Deep Web Shopping: A Case Report.

Cyanide is a product that is known for its use in industrial or laboratory processes, as well as for intentional intoxication. The toxicity of cyanide is well described in humans with rapid inhibition of cellular aerobic metabolism after ingestion or inhalation, leading to severe clinical effects that are frequently lethal. We report the case of a young white man found dead in a hotel room after self-poisoning with cyanide ordered in the deep Web. This case shows a probable complex suicide kit use including cyanide, as a lethal tool, and dextromethorphan, as a sedative and anxiolytic substance. This case is an original example of the emerging deep Web shopping in illegal drug procurement.

Effects of the environment on bone mass: A human taphonomic study.

An experiment was designed using human ribs placed in different environments to document how moisture and temperature affect the bone mass according to the postmortem interval. The bones were defleshed, weighed and partially buried, with some ribs being left unburied as controls. The ribs were weighed daily, and the mass loss was monitored over a period of 90days. The results showed that significant differences in bone mass loss exist between environments, where the bone mass loss was significantly faster in an environment with low moisture content. This mass loss is thought to be primarily associated with the desiccation of the body and then for a greater part, with the atmospheric moisture content. However, the loss of bone mass can also be explained by early alterations in the organo-mineral matrix which were highlighted by Raman spectroscopy method.

Homemade Firearm Suicide With Dumbbell Pipe Triggering by an Air-Compressed Gun: Case Report and Review of Literature.

Firearm suicides are frequent and well described in the forensic literature, particularly in Europe and the United States. However, the use of homemade and improvised firearms is less well described. The present case reports a suicide with an original improvised gun created using an air-compressed pellet gun and a dumbbell pipe. The aims of this study were to describe the scene, the external examination of the corpse, the body scan, and the autopsy; to understand the mechanism of death; and to compare the results with a review of the forensic literature to highlight the epidemiology of homemade firearm use, the tools used for homemade and improvised firearms in suicides versus homicides, and the manners in which homemade firearms are used (homicide or suicide, particularly in complex suicide cases).