New psychoactive substances (NPS) continue to proliferate worldwide, and the pharmacological and toxicological profiles of many remain poorly characterized. Arylcyclohexylamines, derivatives of phencyclidine (PCP) that act as potent N-methyl-D-aspartate (NMDA) receptor antagonists, produce profound dissociative and psychotomimetic effects. 3-Methoxyeticyclidine (3-MeO-PCE), an analog of eticyclidine, has recently appeared on the recreational drug market, yet information on its pharmacology, metabolism, and toxicity are scarce, and only one postmortem blood concentration has been reported to date. We report the death of a 33-year-old man found beneath his apartment window after a fall of 5.8 m. Autopsy confirmed death from multiple traumatic injuries consistent with a fall from height. Toxicological analysis of peripheral blood identified 3-MeO-PCE (80 µg/L), its metabolite 3-HO-PCE (13 µg/L), tramadol (674 µg/L), and ethanol (0.18 g/L). Although trauma was the immediate cause of death, the behavioral effects of 3-MeO-PCE—possibly potentiated by tramadol and alcohol—may have contributed to the manner of death. The disordered state of the apartment, together with the absence of any psychiatric history or suicidal indicators, is compatible with acute agitation or dissociation induced by substance use. This case highlights the potential for serious behavioral disturbances associated with 3-MeO-PCE and adds a new peripheral blood concentration to the limited forensic literature on this compound.
Population affinity estimation remains an important component of the biological profile, particularly in demographically complex populations such as South Africa, where morphological overlap exists among groups. This study evaluated the performance of geometric morphometric (GMM) and inter-landmark distance (ILD) classification approaches derived from an automatic landmarking-based workflow for population affinity estimation in a contemporary South African sample. A total of 474 cranial computed tomography scans representing recorded Black, Coloured, Indian, and White South Africans were analysed. Eighteen three-dimensional craniofacial landmarks were automatically transferred from a reference template to individual cranial surfaces using rigid and non-rigid registration, and nine ILDs were calculated. Mean intra- and inter-observer landmark errors were 0.746 mm and 1.672 mm, respectively. GMM analyses identified significant craniofacial shape differences among all recorded population groups, with the greatest separation between Black and White South Africans and the greatest overlap involving Coloured and Indian South Africans. Cross-validated GMM-based discriminant function analysis achieved an overall classification accuracy of 90.08
Heat-induced deaths create complex challenges in forensic anthropology because thermal exposure causes significant structural and chemical changes in skeletal remains. This systematic review aims to examine heat-induced alterations in human bone and evaluate their importance for forensic anthropological examination in heat-induced contexts. A systematic literature search was conducted with PRISMA guidelines using keywords related to burned bone, forensic anthropology, and fire exposure. Studies focusing on thermally altered human skeletal remains were included, while studies involving animals or archaeological material were excluded. This systematic review shows that thermal exposure produces predictable macroscopic and microscopic changes in bone, including progressive color change, shrinkage, warping, and fracture formation. These changes are influenced not only by temperature but also by oxygen availability, combustion duration, and burning conditions. The findings indicate that the effects of thermal alteration vary according to the component of the biological profile. being assessed, the methods employed, and the degree of skeletal preservation. Trauma examination in burned samples remains possible in some cases, although fragmentation and calcination may obscure diagnostic features. Understanding the mechanisms of heat-induced skeletal alteration improves the examination of burned human remains and supports more reliable forensic anthropological analysis in fire-related investigations.
The prediction of phenotypic traits from DNA, known as forensic DNA phenotyping, or externally visible characteristics, represents a disruptive evolution in contemporary forensic genetics. Although various studies have identified molecular markers associated with facial features, the predictive accuracy of these markers varies significantly between populations due to their genetic background and representative sampling of relevant populations. This systematic review aimed to analyze and synthesize existing studies on forensic DNA phenotyping, evaluate the accuracy of different types of markers, and compare their effectiveness in predicting human facial features. The publication screening was based on the 2020 PRISMA and the search was conducted in three different databases: Scopus, PubMed, and Web of Science, published through May 23, 2025, yielding a total of 301 initial records. After article screening, 28 studies were selected for analysis which were published between 2013 and 2025 and were written in English. A total of thirteen facial feature categories were identified among which facial morphology, presence or absence of freckles, and male-pattern baldness stand out for being associated with a greater number of SNPs. The systematic literature review highlights significant progress in genetic appearance prediction, validating both established assays and in-house panels. While common pigmentary traits achieve high predictive accuracies (> 89–90
Anthropological and forensic investigations often rely on skeletonized human remains as the only piece of evidence for lifestyle, age and post-mortem interval predictions. Recent studies have demonstrated that histological staining has the potential to extend or improve the knowledge that can be extracted from the microstructure of bone. However, histological staining has only been scarcely applied and investigated on (taphonomically altered) human bones, leading to a lack of available methodology. In this study, protocols for the preparation of bone slides for light microscopy and for staining with Hematoxylin and Eosin, Sirius Red & Fast Green, Toluidine Blue, Masson's Trichrome and Azocarmine G were adapted for use on undecalcified, unembedded and unfixed human bone. The final methods were applied to fresh, fridge stored and taphonomically altered bones to observe differences between the staining outcomes, as well as what information can be drawn from staining. The successfully stained bone sections revealed information regarding morphology and the collagen content of the bone based on the affinity of the stains to targeted microstructures in the bone or to the collagen molecules respectively. These bone preparation and staining methods are a valuable step towards the adaptation of other stains for human bone, and ultimately towards the development of uniformized histological methods to support forensic-anthropological investigations.
While thermal injuries caused by heat are well-recognized in cases of child maltreatment, injuries resulting from cold exposure (hypothermia and local cold injury) appear to be rare in this context. Although hypothermia and cold injuries related to child abuse and neglect pose a significant threat to affected children, systematic data on this topic are currently lacking. The goal of this article was to better understand the frequency, forms and contexts of child maltreatment involving cold exposure. We conducted a narrative review with systematic elements based on a PubMed search and an additional non-systematic search of German language publications in Google Scholar. Two reviewers independently screened abstracts, applying inclusion and exclusion criteria. Full texts of potentially relevant studies were then assessed to determine final inclusion. The search yielded 192 search results. Additionally, 4 publications were found in a non-systematic German language search. 22 publications were ultimately included in the review. The limited number of publications restricts generalizability. Hypothermia and local cold injuries in child maltreatment mainly affect children under six years and may present in varied forms and scenarios. Underreporting due to non-recognition and potential publication bias should be considered, and differential diagnoses must be carefully assessed in practice.
To evaluate the diagnostic accuracy of postmortem blood procalcitonin (PCT) in identifying sepsis as a cause of death through a systematic review and meta-analysis. A comprehensive literature search of PubMed, Scopus, ScienceDirect and Cochrane library was performed. Studies evaluating postmortem PCT levels were included. Data were extracted from all studies for diagnostic accuracy parameters and study quality was assessed. Pooled sensitivity and specificity were estimated. Diagnostic accuracy was further evaluated using a Reitsma bivariate random-effects model and hierarchical summary receiver operating characteristic (HSROC) analysis. 11 studies met the inclusion criteria, of which eight studies provided sufficient data for pooled analysis. Most studies used peripheral (femoral) blood, while some used different matrices such as pericardial fluid, cerebrospinal fluid, and aqueous humour. The pooled analysis of postmortem blood-based studies demonstrated a sensitivity of 91.0
Forensic anthropology is the application of the knowledge and methods of physical anthropology in a medico-legal context. The unknown skeletal remains are examined to assess and assist in identifying them. When conventional methods fail, facial reconstruction is often the last recourse attempted for identifying an unknown skull. A partial nasal bridge, a small anterior nasal spine, with a large hollowed space are all that are present on a skull representing the nose. Hence, it is a challenging task for a forensic scientist to build a soft nose based on the scant available bony structure. The present study was conducted on computed tomography (CT) images of 409 individuals (226 males and 183 females). Seventeen metrical parameters, including soft tissue depths, distances and angles, were measured. Regression models were constructed to assess the various dimensions of the nose. The study also included 40 morphological attributes to identify the most frequent nasal features in the studied population. The morphological study suggests two methods. In the absence of nasal bones, it recommends using the "most frequent nasal features" method. For partially broken or intact nasal bones, the method of "characteristic association" may be adopted. The nasal framework was constructed with metrical parameters and later refined morphologically. This will increase the likelihood that a reconstructed skull resembles the antemortem face.
Ethanol is one of the most widely used psychoactive substances worldwide and one of the most frequently investigated substances in forensic toxicology laboratories. Choosing the most appropriate biological sample for its screening is extremely important in forensic autopsy, mainly to differentiate alcohol consumed before death from alcohol formed post-mortem, as well as to reduce the risk of ethanol loss through oxidation or metabolism. Therefore, this study presents an integrative review of recent literature on the collection, storage, and availability of biological matrices collected during forensic autopsy for detecting alcohol and its metabolites, considering the post-mortem condition and decomposition status of the body. The Scopus, Web of Science, and PubMed databases were searched, with searches limited to English-language articles from the past 10 years, excluding reviews and meta-analyses. The language and time restrictions were adopted to ensure methodological consistency, linguistic homogeneity, accessibility for an international readership, and focus on recent evidence relevant to current forensic toxicology practice. After excluding duplicates, the initial screening identified 414 articles. After analyzing the titles, abstracts, and full texts, articles with unavailable full text, incorrect study design, incorrect publication type, or off-topic content were excluded. From a detailed analysis of the 54 definitively included articles, data were collected on study objectives, main results, recommendations for collection and storage, and potential biomarkers. In general, it is recommended to analyze multiple matrix types and biomarkers, collect samples in duplicate, refrigerate or freeze them depending on the storage time, use preservatives, and provide adequate staff training. Protocols that provide greater clarity between results and more faithfully reflect real conditions are relevant to investigations and can prevent or detect alterations in ethanol levels in biological samples, thereby ensuring the validity, integrity, and traceability of evidence from its collection to its presentation in court.
Postmortem diagnosis of hypothermia remains challenging even in the era of advanced virtual and molecular diagnostic technologies. Although numerous morphological findings have been described in deaths due to cold, only two—Wischnewsky spots and cold erythema (Keferstein’s sign)—have traditionally been regarded as reliable autopsy indicators of fatal hypothermia. More than a century after their first description, additional supportive findings, including hemorrhages within core skeletal muscles and the inner knee sign, have expanded the spectrum of autopsy features associated with fatal cold exposure. Reactive splenic contraction, characterized by reduced splenic volume and weight, capsular wrinkling, and red-pulp anemization, has so far been reported only sporadically and has never been systematically validated in a larger human series. The present prospective controlled autopsy study investigated the occurrence and diagnostic significance of reactive splenic contraction in 54 fatalities due to hypothermia and 170 controls. The reactive splenic contraction triad was identified in 48 of 54 hypothermia cases (88.9
Due to the nearly identical genomic sequences of monozygotic twins (MZTs), traditional autosomal Short Tandem Repeat (STR) genotyping methods are unable to distinguish between them. Epigenetic factors exhibit potential value in forensic science. Dynamic epigenetic factors may be influenced by genetic traits or postnatal environmental factors. This study focused on long non-coding RNAs (lncRNAs), and aimed to comprehensively screen for differentially expressed lncRNAs between MZTs using next-generation sequencing (NGS). Additionally, the consistency of these differentially expressed lncRNAs across multiple twin pairs was evaluated by quantitative PCR (qPCR). Through lncRNA sequencing (lncRNA-seq), a total of 12,547 expressed lncRNAs were identified in four pairs of MZTs, revealing 31 commonly differentially expressed lncRNAs. In the PCR preliminary experiment, 11 lncRNAs showed stable amplification effects and high specificity, providing a basis for further validation, including MSTRG.30665.2, ENST00000414030, MSTRG.73422.1, MSTRG.87582.24, MSTRG.100816.5, MSTRG.54028.15, MSTRG.16169.7, MSTRG.98182.1, MSTRG.68271.29, MSTRG.63897.85, and MSTRG.16294.1. Subsequently, qPCR confirmed the identification ability of these lncRNAs. The identification ability of these 11 lncRNAs varies, with MSTRG.73422.1 having the strongest discrimination power. The forensic applicability assessment showed that five lncRNAs exhibited strong resistance to degradation under specific environmental conditions and robust longitudinal stability. Their expression levels did not significantly decrease after storage at room temperature for up to 180 days or following 10 freeze-thaw cycles. In summary, our study reveals the potential application of lncRNAs in the identification of MZTs.
Waterhouse–Friderichsen syndrome (WFS) represents a fulminant septic condition characterized by bilateral adrenal hemorrhage and rapid clinical deterioration. Although classically associated with meningococcal infection, fatal cases occur across multiple infectious etiologies and heterogeneous clinical contexts. In forensic practice, diagnosis frequently relies on post-mortem interpretation rather than clinical recognition, making consistent pathological assessment essential. A systematic review was conducted according to PRISMA guidelines to identify autopsy-confirmed fatal cases consistent with WFS. PubMed/MEDLINE and Scopus were searched from database inception to 21 February 2026, with a supplementary search conducted in Google Scholar. Eligible studies included fatal human cases with bilateral adrenal hemorrhage documented at autopsy and sufficient pathological and/or microbiological data. Data were extracted on clinical course, gross and histopathological findings, microbiological investigations, pathogens, and predisposing factors. Owing to marked heterogeneity of case-level reporting, qualitative synthesis and semi-quantitative descriptive analysis were performed. Sixty-five studies, encompassing 86 individual fatal cases, met the inclusion criteria. Bilateral adrenal hemorrhage consistently occurred within a broader systemic septic pattern characterized by multiorgan congestion, hemorrhagic manifestations, and variable disseminated intravascular coagulation-related changes. Histopathological and microbiological findings were heterogeneous, with frequent discrepancies between culture-based and molecular results and recurrent diagnostic limitations related to post-mortem microbiology. The synthesis of evidence supports interpreting WFS as a recurrent forensic-pathological pattern rather than a single morphological diagnosis. Based on key findings identified across cases, practical forensic frameworks were developed to support autopsy interpretation, microbiological sampling, and differential diagnosis. Integration of morphology, histopathology, microbiology, and circumstantial data may improve consistency and reproducibility in the medico-legal evaluation of sudden septic deaths.
Traumatic brain injury (TBI) is a major cause of death and long-term impairment in children and adolescents and is relevant to both clinical and forensic medicine. The aim of this study was to investigate associations between age, mechanism of injury and injury patterns in fatal pediatric TBI cases. In a retrospective observational study, 96 fatal cases in children and adolescents aged 0–18 years were analyzed over a 20-year period. A bimodal age distribution was observed, with peaks in early infancy and adolescence. Traffic accidents predominantly affected older children and were associated with complex cranial and facial fractures as well as subarachnoid and intraventricular hemorrhage (SAH, IVH). In contrast, abusive head trauma (AHT) mainly occurred in children aged two years or younger and was characterized by a higher prevalence of subdural hemorrhage (SDH) and an almost complete absence of cranial fractures. Age- and mechanism-specific injury patterns may help improve forensic evaluation in unclear cases and provide useful insights for prevention and clinical and forensic decision-making.
Forensic entomology can be pivotal for minimum postmortem interval (PMImin) estimation. Case-based studies can support forensic investigations and guide laboratory research, but systematic data from real cases in China remain limited. In this study, 65 forensic entomology cases collected from 2017 to 2024 in two regions of China, Beijing and Hunan, were analyzed to characterize necrophagous insect occurrence patterns and larval gut microbiota. A total of 30 colonizing insect species from 11 families were identified. Lucilia sericata, Chrysomya megacephala, and Sarcophaga crassipalpis were the most frequently recorded species. Diptera, especially Calliphoridae and Sarcophagidae, were mainly associated with shorter postmortem intervals, whereas coleopteran taxa were more often observed in longer-interval cases. In addition, insect occurrence showed clear geographical distribution and seasonality: L. sericata and S. crassipalpis predominated in Beijing cases, whereas C. megacephala and Sarcophaga peregrina were more frequent in Hunan cases. Overall species diversity was highest in summer, whereas only Calliphora vicina and Aldrichina grahami were observed in winter. 16S rRNA gene sequencing of larval gut microbiota revealed a stable core microbiome across diverse cases and species, including Ignatzschineria, Clostridium, Peptostreptococcus, Vagococcus, and Peptoniphilus. This study provides regional forensic entomology reference data and characterizes the gut microbiota of fly larvae collected directly from human remains in police casework, offering a useful basis for future forensic investigations and laboratory studies.
Forensic Anthropology applies biological characteristics in legal contexts to estimate the biological profile of an unknown individual. In this regard, teeth stand out due to their resistance and ability to preserve valuable information over time. More specifically, microstructural records present in dental enamel, in the form of incremental markings known as Retzius striae, provide data on dental development and, consequently, on the individual. The objective of this systematic review was to assess the applicability of Retzius striae in Forensic Anthropology. A search was conducted using topic-related keywords across six different databases in Portuguese, Spanish, and English. Studies were selected according to predefined inclusion and exclusion criteria. The methodological quality and risk of bias of the included studies were assessed. As a result, seven scientific studies addressing Retzius striae in contemporary human samples were identified. All investigations evaluated the striae using histological methods. Furthermore, all studies demonstrated a low risk of bias. Retzius striae have been investigated as markers for estimating age, sex, stature, and even body weight; however, the available evidence remains limited. Although they show potential, all these applications require further validation. Therefore, additional standardized and representative primary studies are necessary to ensure reliable and safe application in the forensic context.
Rib fractures are among the most common injuries in traffic accidents. Accurate evaluation of rib fracture age is important as it assists forensic experts in reconstructing injury processes and provides objective technical support for subsequent judicial judgments. Routine diagnostic methods sometimes cannot meet forensic requirements for fracture-age estimation. Technetium-99m-methyldiphosphonate (99mTc-MDP), a radiotracer administered intravenously, binds dynamically to hydroxyapatite involved in fracture healing. 99mTc-MDP uptake increases in areas with abundant blood flow and active osteoblast repair at the fracture site, offering potential for estimating fracture age. In this study, 99mTc-MDP uptake was measured using the semi-quantitative target-to-non-target ratio (T/NT) through single-photon emission computed tomography. A total of 151 rib fractures were included from 55 individuals. Fracture age ranged from 3 to 491 days and was divided into three groups (within 30 days, 30–60 days, and over 60 days). T/NT values differed significantly among groups using Mann-Whitney U tests (P < 0.0001). The correlation coefficient (R2) between fracture age and T/NT was 0.36 (P < 0.0001). Six machine-learning models were then constructed, among which the support vector machine model had the best performance with area under the curve (AUC) of 0.91, F1-score of 0.78, accuracy of 0.80, and recall of 0.79 when identifying the three groups in the independent testing set. This study offers a novel perspective for estimating rib fracture age using functional imaging, which may provide essential information for case reconstruction and serve as a useful complementary tool in forensic bone trauma analysis.
In sexual assault investigations, the rapid and accurate localization of biological traces on clothing is important for guiding evidence collection and preserving forensic material. Multispectral imaging systems provide a non-invasive optical approach for visualizing body fluids that may be difficult to detect under conventional lighting conditions. Seven biological fluids—venous blood, menstrual blood, semen, vaginal secretions, urine, saliva, and sweat—were applied to cotton and synthetic fabrics of different colors and textures. After air-drying, the samples were examined using three portable multispectral imaging devices: Forenscope Multispectral SAFE/Colposcopy, Safe Pro Smartphone, and Mobile Multispectral Tablet. Stain visibility was initially assessed using a predefined ordinal visual scoring system. In addition, post hoc quantitative image analysis was performed on exported image files by calculating signal intensity, background intensity, contrast, and signal-to-noise ratio (SNR). On light-colored fabrics, most biological fluids were readily detectable under white or visible light. On dark fabrics, infrared illumination provided improved stain-background separation, particularly for blood and semen. Semen, vaginal secretions, and saliva showed visible fluorescence under blue light in the 440–470 nm range combined with an orange filter. Quantitative analysis demonstrated significant differences in contrast and SNR according to imaging device and fabric color, whereas fabric type did not show a significant effect. Biological fluid type significantly affected contrast, but not SNR. The detectability of biological fluids on clothing depends on fluid type, fabric characteristics, and spectral imaging conditions. Portable multispectral imaging systems may support the rapid, non-destructive localization of biological traces in forensic examinations. However, the present study should be interpreted as an exploratory evaluation under controlled conditions, rather than as diagnostic validation or evidence of superiority over conventional forensic detection methods.
A man in his early twenties had been hospitalized due to acute nitrous oxide intoxication, during which a vitamin B12 deficiency and anaemia were also diagnosed. Two weeks later he was found dead in his apartment. Twenty empty nitrous oxide cartridges were found next to the body, and according to his mother, he had been using nitrous oxide excessively for approximately two months. The autopsy revealed a fatal pulmonary embolism secondary to deep vein thrombosis, and toxicological analysis detected nitrous oxide in low concentrations in the blood. Regarding the pulmonary embolism, a coagulation disorder attributable to suspected hyperhomocysteinemia in the setting of vitamin B12 deficiency could be considered a contributory factor – this deficiency, in turn, may possibly be related to excessive nitrous oxide consumption over several weeks. Obesity and nicotine abuse were present as additional risk factors for thromboembolic events. Reapeted excessive nitrous oxide consumption can induce profound metabolic disturbances, including functional vitamin B12 deficiency and associated biochemical derangements. In cases of fatal thromboembolic events in young individuals, this possibility should be taken into account. Toxicological analysis may demonstrate recent nitrous oxide exposure and support the hypothesis, but it cannot establish the underlying biochemical pathology.
