[Features of detection and interpretation of intravital and postmortem changes according to the results of traditional X-ray and X-ray computed tomography of objects from historical graves and artefacts]. / Osobennosti vyyavleniya i interpretatsii prizhiznennykh i posmertnykh izmenenii po rezul'tatam traditsionnoi rentgenografii i rentgenovskoi komp'yuternoi tomografii ob"ektov iz istoricheskikh zakhoronenii i artefaktov.

OBJECTIVE: To study emergence mechanism, physical nature, pattern of intravital and postmortem changes of biological and non-biological objects originated in the period from 1550 to 1918 yr. using traditional X-ray and X-ray computed tomography. MATERIAL AND METHODS: The relics of Saint Macarius the Roman of Novgorod, the remains of the First Reverend of the Resurrection Novodevichy Convent in Saint Petersburg Mother Superior Theophania, damages on the chair leg on which Tsesarevich Alexey sat during the shooting of Russian Emperor Nicholas II, his family and entourage in 1918 in Yekaterinburg were stidued. RESULTS AND CONCLUSION: The application of highly informative methods of traditional X-ray and X-ray computed tomography of biological and non-biological objects showed their high informativity and allowed to correctly interpret the emergence mechanism, physical nature, pattern of intravital and postmortem changes of skeleton bones and historical artefact (chair legs) originated long ago. The necessity of special professional training and advanced training of experts in forensic radiology to prevent possible diagnostic and expert errors has been substantiated.

MEDICO-LEGAL APPLICATIONS OF FRACTURE HEMATOMA: REVIEW.

This review aimed to elucidate the critical role of fracture hematoma in forensic medicine, with a specific focus on its utility in differentiating antemortem from postmortem fractures. The study seeks to provide a comprehensive synthesis of current knowledge on the subject, highlighting the biological and medico-legal implications of fracture hematoma analysis in forensic investigations. A systematic review of literature was conducted, encompassing various scientific databases including PubMed, Scopus, and Web of Science, focusing on studies published from 2000 to 2024. The search employed keywords such as "fracture hematoma," "antemortem fractures," "perimortem fractures" and "postmortem fractures," among others, to explore relevant data. Selected studies were scrutinized based on their relevance, the presence of substantial data on fracture hematoma, and their contribution to forensic analysis. The review underscores the significance of fracture hematoma as an indicator of antemortem injuries, revealing that active blood circulation at the time of injury facilitates hematoma formation. Detailed analyses within the selected studies illustrate the interplay of cellular and molecular dynamics within fracture hematomas, emphasizing the roles of cytokines, particularly IL-6, and cellular constituents in the healing process. Fracture hematoma analysis emerges as a vital forensic tool in establishing the vitality of bone fractures, enhancing the accuracy of forensic assessments. However, the review also acknowledges the challenges posed by individual healing variability and postmortem changes, suggesting a need for further research to refine the interpretative frameworks used in forensic hematoma analysis.

Volabolomic Fingerprinting for Post-Mortem Interval Estimation: A Novel Physiological Approach.

Death is a multifaceted process wherein each individual cell and tissue has a metabolic homeostasis and a time of functional cessation defined by the dying process as well as by intrinsic and extrinsic factors. Decomposition is physiologically associated with the release of different types of volatile organic compounds (VOCs), and these form volaboloma mortis. The main purpose of this study was to record the volabolomic fingerprint produced by volatile molecules during the physiological decomposition process of human tissue and muscle cells. The volatile chemical signature has important implications for an open issue in forensics and pathology, namely the estimation of the postmortem interval (PMI), which decreases in accuracy with the passage of time. Volatile metabolites emitted from human tissues and muscle cells at 0, 24, 48, and 72 h were recorded in real time with an electronic nose sensor device. The key findings were the continuous sampling of VOCs emitted from tissues and cells. These showed a common behavior as time progressed; particularly, after 48 h the distributions became dispersed, and after 72 h they became more variable. Volabolomic fingerprinting associated with time progression relevant to the study of PMIs was reconstructed. Additionally, there may be broader applications, such as in dog training procedures for detecting human remains, and perhaps even for studying scavenger and insect attractants.

Postmortem chondrocyte viability in porcine articular cartilage: Influence of time, temperature, and burial under winter conditions.

The aim of the present study was to investigate the effects of time, temperature, and burial in a natural environment on the viability of chondrocytes in porcine femoral condyles using confocal laser scanning microscopy. Hind trotters from 10 pigs were buried or left unburied. Samples were collected daily and stained with a combination of vital dyes (calcein-AM and ethidium homodimer-1). The chondrocytes showed an intense staining corresponding to their vitality. In the first 3 days, viability decreased slowly and showed no statistical difference between buried and unburied samples. After the first 3 days, it decreased rapidly, with the viability of the buried samples being 66% on day 4, decreasing to 25% on day 8 and to 16% on day 10, while in the unburied samples it decreased to 43% on day 4, 13% on day 8 and 5% on day 10. Our results indicate a time, temperature, and burial dependent decrease in chondrocyte viability and suggest the use of chondrocyte viability as a marker for estimating PMI in both the natural environment and in animals, as well as its potential use in humans.

The influence of altitude on the abundance and occurrence of species of the family Calliphoridae (Diptera) in corpses in a Mediterranean area.

A carcass is defined as a temporary resource that can support high levels of diversity compared to other resources. The level of diversity often depends on the environmental conditions in which the corpse is found. Calliphoridae (Diptera) are the most important necrophagous insects used in forensic investigations because this family is common, widespread, abundant, and usually the first to colonize a corpse. However, understanding the processes responsible for variation in calliphorid species abundance and richness along gradients in ecology remains a key challenge. In this study, we analyzed the influence of altitude on the abundance and diversity of Calliphoridae species in corpses at three different altitudes (400, 900, and 1500 m a.s.l.) in a mountainous area of southeastern Spain. The results revealed a gradient of decreasing abundance with increasing altitude. An altitudinal substitution of species was observed, with Chrysomya albiceps being found at the lowest altitude and Calliphora vicina at the highest altitude. Seasonal variation was also noted, with Ch. albiceps being the dominant species in the warmer months and C. vicina being the dominant species in winter. Our results confirm the importance of Calliphorids as seasonal, altitudinal, and environmental indicators, given the wide distribution and abundance of this family. This information is of great interest on the interpretation of data in forensic practice.

A primary study of ethanol production in postmortem liver and muscle tissue of rats.

OBJECTIVE: To study the characteristics of postmortem ethanol production and its relation with alcohol congeners in postmortem rat liver and muscle tissues. METHOD: Postmortem liver and muscle tissues in Sprague-Dawley rats, from postmortem time interval (PMI) day 0-20, were analyzed via headspace gas chromatograph flame ionization detection to observe production of postmortem ethanol and 5 selected alcohol congeners. RESULT: 1. Putrid ethanol production increased gradually to a peak and then decreased with the prolongation of PMI; 2. Acetaldehyde, 1-propanol, and 3-methyl-butyraldehyde were produced along with postmortem ethanol; 1-butanol was only detected from day 11-20; 3. The concentrations of acetaldehyde, 1-propanol and 3-methyl-butyraldehyde was related with ethanol production. Fifteen mathematical models were constructed for putrid ethanol production based on acetaldehyde, 1-propanol, and 3-methyl-butyraldehyde. CONCLUSION: A peak in postmortem ethanol production was identified. The production trends of acetaldehyde, 1-propanol, and 3-methyl-butyraldehyde in the liver, and of 1-propanol in muscle, were consistent with those of ethanol, and could potentially to be used as biomarkers of postmortem ethanol production. Further human samples and data analysis are needed to verify this.

The Role of Protein Degradation in Estimation Postmortem Interval and Confirmation of Cause of Death in Forensic Pathology: A Literature Review.

It is well known that proteins are important bio-macromolecules in human organisms, and numerous proteins are widely used in the clinical practice, whereas their application in forensic science is currently limited. This limitation is mainly attributed to the postmortem degradation of targeted proteins, which can significantly impact final conclusions. In the last decade, numerous methods have been established to detect the protein from a forensic perspective, and some of the postmortem proteins have been applied in forensic practice. To better understand the emerging issues and challenges in postmortem proteins, we have reviewed the current application of protein technologies at postmortem in forensic practice. Meanwhile, we discuss the application of proteins in identifying the cause of death, and postmortem interval (PMI). Finally, we highlight the interpretability and limitations of postmortem protein challenges. We believe that utilizing the multi-omics method can enhance the comprehensiveness of applying proteins in forensic practice.

Post-mortem formation of ethanol: Is 1-propanol a reliable marker? A proof-of-concept study using an in vitro putrefactive environment setup.

Ethanol is the psychoactive substance identified most frequently in post-mortem specimens. Unfortunately, interpreting post-mortem ethanol concentrations can be difficult because of post-mortem alcohol redistribution and the possibility of post-mortem alcohol neogenesis. Indeed, in the time interval between death and sample collection, the decedent may be exposed to non-controlled environments for an extended period, promoting microbial colonization. Many authors report that in the presence of carbohydrates and other biomolecules, various species of bacteria, yeast, and fungi can synthesize ethanol and other volatile substances in vitro and in vivo. The aim of this study was to study the impact of several variables on microbial ethanol production as well as develop a mathematical model that could estimate the microbial-produced ethanol in correlation with the most significant consensual produced higher alcohol, 1-propanol. An experimental setup was developed using human blood samples and cadaveric fragments incubated under strictly anaerobic conditions to produce a novel substrate, "cadaveric putrefactive blood" mimicking post-mortem corpse conditions. The samples were analyzed daily for ethanol and 1-propanol using an HS-GC-FID validated method. The formation of ethanol was evaluated considering different parameters such as putrefactive stage, blood glucose concentration, storage temperature, and storage time. Statistical analysis was performed using the Mann-Whitney non-parametric test and simple linear regression. The results indicate that the early putrefactive stage, high blood glucose concentration, high temperature, and time of incubation increase microbial ethanol production. In addition, the developed mathematical equation confirms the feasibility of using 1-propanol as a marker of post-mortem ethanol production.

Morphometric analysis for determination of larval instars in Dermestes frischii Kugelann and Dermestes undulatus Brahm (Coleoptera: Dermestidae).

Dermestes frischii Kugelann, 1792 and Dermestes undulatus Brahm, 1790 are the most abundant species worldwide at outdoor or indoor crime scenes during the dry and skeletal stages of decomposition. The attribution of larval age in these beetles is problematic due to the variable number of instars, which is influenced by environmental factors. In this study, a morphometric approach was used to look for potential morphological features as evidence of larval stages. Breeding and monitoring were performed for both species in an incubator with a preset temperature of 28°C ± 0.5 without a photoperiod. Morphometric measurements were made on 10 larvae per instar for each species using length, width, and thickness parameters. Linear discriminant analysis was then used to generate decision boundaries that clearly separated larval stages. The cross-validation procedure demonstrated that the morphometric approach successfully discriminated adjacent larval stages in both species with high values of sensitivity and specificity. This less-invasive approach could improve the ability to estimate minPMI in forensic studies of Dermestidae beetles. Future studies may extend this approach to other species and establish good practices for collecting and storing specimens for morphometric analysis.

Evaluation of porcine decomposition and total body score (TBS) in a central European temperate forest.

The total body score (TBS) is a visual scoring method to scale the succession of decomposition stages. It compares decomposition between cadavers, to connect it with external taphonomic factors and estimate the post-mortem interval. To study decomposition in various climatic environments, pigs are often used as human proxies. Currently, there is one TBS system by Keough et al. (J Forensic Sci. 2017;62:986) for surface-deposited domestic pigs, coming from South Africa. Our study aims to evaluate this method and analyze porcine decomposition in Central Europe to inform forensic research and casework. We conducted an experiment studying six 50 kg pig carcasses in a temperate Swiss forest. Three observers documented decomposition patterns and rated the decomposition stages from photographs based on the porcine TBS model by Keough et al. (J Forensic Sci. 2017;62:986). We documented discrepancies between the carcass decomposition of our specimens and those in the South African study, especially related to the high insect activity in our experiment. Furthermore, we noted factors complicating TBS scoring, including rainfall and scavengers. The agreement between TBS observers from photographs was in the highest agreement category apart from one "substantial agreement" category. Our study is the first in Europe to systematically test the Keough et al. (J Forensic Sci. 2017;62:986) method. The results evidence that regional adaptations are required to be applicable for other environments. We present a modified approach based on experimental observations in a Swiss temperate forest. The identification of regional decomposition patterns and drivers will inform future taphonomy research as well as forensic casework in comparable contexts in Central Europe.

Ant bite artifacts in a case of hanging.

Artifacts produced by postmortem animal scavenging are commonly encountered at autopsy. Knowledge of the pattern of artifacts produced by postmortem animal scavenging is essential for the correct interpretation of the autopsy finding. In household deaths, such artifacts are usually caused by domestic animals and by small insects such as flies, ants, beetles, etc. Ants are one of the early scavengers which feed on the dead bodies. The artifacts produced by the postmortem ant activity are usually superficial and non-bleeding type. Rarely, in the congested body regions and areas of marked hypostasis, postmortem bleeding artifacts due to ant bites are possible. In the reported case of hanging, such postmortem bleeding artifacts were present over both the legs, predominantly over the left leg. Typical ant bite lesions in the form of superficial excoriations were also present over the peri-ligature area, over and around both nipples and over the lower part of the abdomen. Morphologically, postmortem bleeding artifacts produced by ant bites exhibit four patterns: droplet pattern, stripe pattern, pool pattern, and mixed pattern. In this case, a mixed pattern (droplet pattern and stripe pattern) of postmortem bleeding artifacts was observed. The presence of postmortem bleeding artifacts over the lower limbs was attributed to the pooling of the blood due to suspension of the body, followed by passive escape of blood due to ant bites. The possibility of such artifacts produced by ant bites should be considered when the origin of the lesion is unclear.

New perspective for Calpain-Mediated regulation of meat Quality: Unveiling the impact on mitochondrial pathway apoptosis in post-mortem.

While calpain's role in myofibrillar protein degradation is well-established, its impact on post-mortem apoptosis remains fully elucidated. This study aimed to examine how calpain influences the mitochondrial apoptotic pathway in post-mortem muscle cells and assess its potential impact on chicken tenderness. The findings indicate that the calpain inhibitor treatment could decelerate the rate of lysosome destruction in post-mortem chicken, which is a crucial factor in delaying the mitochondrial apoptotic pathway. Subsequently, this inhibition enhanced the mitochondrial membrane's stability and suppressed the apoptosis-inducing factor Cyt c release into the sarcoplasm. The Western blot results in a greater myofibrillar protein degradation degree in the caspase inhibitor samples compared to the calpain inhibitor samples. Interestingly, the two groups had no significant difference in shear force. Based on these reasons, a novel perspective was introduced in this paper: Calpain could affect the change in meat tenderness by regulating mitochondrial apoptosis in the post-mortem period.

geoFOR: A collaborative forensic taphonomy database for estimating the postmortem interval.

Accurately assessing the postmortem interval (PMI), or the time since death, remains elusive within forensic science research and application. This paper introduces geoFOR, a web-based collaborative application that utilizes ArcGIS and machine learning to deliver improved PMI predictions. The geoFOR application provides a standardized, collaborative forensic taphonomy database that gives practitioners a readily available tool to enter case information that automates the collection of environmental data and delivers a PMI prediction using statistically robust methods. After case submission, the cross-validating machine learning PMI predictive model results in a R² value of 0.82. Contributors receive a predicted PMI with an 80% confidence interval. The geoFOR database currently contains 2529 entries from across the U.S. and includes cases from medicolegal investigations and longitudinal studies from human decomposition facilities. We present the overall findings of the data collected so far and compare results from medicolegal cases and longitudinal studies to highlight previously poorly understood limitations involved in the difficult task of PMI estimation. This novel approach for building a reference dataset of human decomposition is forensically and geographically representative of the realities in which human remains are discovered which allows for continual improvement of PMI estimations as more data is captured. It is our goal that the geoFOR data repository follow the principles of Open Science and be made available to forensic researchers to test, refine, and improve PMI models. Mass collaboration and data sharing can ultimately address enduring issues associated with accurately estimating the PMI within medicolegal death investigations.

Parasitic and predatory behavior of Alysia manducator (Hymenoptera: Braconidae) on blow fly larvae feeding on an adult pig carcass in the Western Cape Province of South Africa: preliminary observations and forensic implications.

Wasps are part of the entomofauna associated with vertebrate carrion. They are known to parasitize and prey on specific life stages of insect hosts such as eggs, larvae, pupae, and/or adults associated with vertebrate carrion. However, reports of parasitic behavior of wasps on carrion-associated insect life stages and their possible forensic implications are non-existent in the Western Cape Province of South Africa. This study is part of ongoing research investigating the entomofauna and their pattern of succession on an adult pig carcass in Cape Town, South Africa. During this study, the parasitic wasp Alysia manducator was noted parasitizing and preying on blow fly larvae associated with the decomposing carcass. The arrival of A. manducator coincides with the occurrence of blow fly eggs and/or larvae on the carcass. These wasps were seen in close association with the eggs and larvae of blow flies on various parts of the carcass and some wasps were seen dragging fly larvae attached to their ovipositors away from one part of the carcass to another. Some A. manducator were also observed walking over several larvae on the carcass while exhibiting a stabbing behavior presumably in search of a host for oviposition. We suggest that the observations recorded in this study are of considerable forensic importance as the dragging effect and predatory and stabbing behavior exhibited by A. manducator could potentially disrupt the feeding and development of the fly larvae on the carcass. This could subsequently alter the process of carcass decomposition and/or affect minimum post-mortem interval estimations.

Key for the identification of third instar larvae of African blowflies (Diptera: Calliphoridae) of forensic importance in death investigations.

Blowfly larvae are the insects primarily responsible for the active stage of decomposition of exposed vertebrate remains and are the most frequently collected entomological evidence during forensic investigations of death. The necrophagous calliphorids in continental Africa that consistently develop on large vertebrate carrion include 11 species belonging to four genera: Calliphora, Chrysomya, Hemipyrellia and Lucilia. Most of these species are widespread in Africa and frequently reported on large animal carcasses and carrion and human corpses. A few keys have been compiled for identification of their third instar larvae, but none of them covers the complete set of taxa. Therefore, we provide a new comprehensive key with original illustrations of all taxonomically significant characters. The key is based on characters that should be easily observable even in poorly equipped local laboratories and is a reliable taxonomic tool for material collected in either urban or rural areas where synanthropic species predominate. However, it should be used with some caution in areas with relatively pristine natural habitats, where additional carrion-breeding species may occur. The publication of the key will significantly facilitate both medical and forensic entomological research and practice in Africa.

The relationship between postmortem interval and protein changes in mice.

Postmortem interval (PMI) estimation is important for forensic pathological autopsy. It has been reported that there is a correlation between certain protein changes in cadavers and PMI. However, no specific protein(s) has been used to determine the PMI so far. In this study, the total protein contents of mouse liver and spleen at different time of death were measured. The data showed that they were negatively correlated with the PMI. The degradation of ß-actin was found to be positively correlated with the PMI in the liver. Additionally, proteomic technique was used to study the changes of protein expression related to PMI in the liver of mice. By using Two-dimensional electrophoresis, the expressions of four proteins were found to be significantly decreased and those of other three proteins were unchanged with the increase of PMI. Among the seven proteins, six were identified with peptide mass fingerprinting using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. The three altered proteins were SBP2, ENOA, ALDH2 and three unchanged ones were 3HAO, TPIS, CATA, respectively. In the future, those unchanged proteins could be used as internal references to more accurately and reliably infer the time of death by assessing the level of changed proteins.

Bone citrate content as an indicator of post-mortem interval.

The citrate levels in the bone start to decrease after death. Therefore, it may be possible to estimate the post-mortem interval if the citrate content decreases at regular and predictable intervals. To verify this, it was used twelve fresh porcine tibiae and femora, fifteen recently buried porcine bones, two samples of human bones curated from the anatomical teaching collection from Bournemouth University, UK (never been buried) and fifteen samples of archaeological human bones from Saint Augustine the Less, Bristol, UK. The results obtained in this study align together with the data available in the literature, with higher citrate content in the fresh porcine bones and lower in the archaeological samples, however, a statistical significance was not found. Moreover, the formula used to estimate the time since death largely overestimated the known PMI. Further research should be done with a larger sample and a new formula to estimate the post-mortem interval is required.

Bridging the gap between decomposition theory and forensic research on postmortem interval.

Knowledge of the decomposition of vertebrate animals has advanced considerably in recent years and revealed complex interactions among biological and environmental factors that affect rates of decay. Yet this complexity remains to be fully incorporated into research or models of the postmortem interval (PMI). We suggest there is both opportunity and a need to use recent advances in decomposition theory to guide forensic research and its applications to understanding the PMI. Here we synthesise knowledge of the biological and environmental factors driving variation in decomposition and the acknowledged limitations among current models of the PMI. To guide improvement in this area, we introduce a conceptual framework that highlights the multiple interdependencies affecting decay rates throughout the decomposition process. Our framework reinforces the need for a multidisciplinary approach to PMI research, and calls for an adaptive research cycle that aims to reduce uncertainty in PMI estimates via experimentation, modelling, and validation.

Recognition of the Presence of Bone Fractures Through Physicochemical Changes in Diagenetic Bone.

Much research has focused on attempting to understand the drivers of bone diagenesis. However, this sensitive process is easily influenced by various factors, particularly the condition of the remains (i.e., whether they have been subjected to trauma). Previous research demonstrates that trauma can influence soft tissue decomposition, yet to date, no studies have looked at how bone fractures could affect bone diagenesis. To address this gap, two short timescale studies were conducted to investigate the influence of bone fractures on the physicochemical composition of disarticulated, partially fleshed animal remains. Disarticulated porcine bones were either fractured using blunt force or sharp force whilst fresh (producing perimortem damage), at 60 days producing postmortem damage (postmortem interval (PMI)), or left intact and left outside for up to 180 days post-fracture/240 days PMI. Retrieved bone sections were then analyzed for physicochemical differences using non-destructive methods, i.e., scanning electron microscopy energy dispersive spectroscopy and Fourier transform infrared spectroscopy with attenuated total reflectance. It was hypothesized that differences would be found in the physicochemical composition between the bones with fractures and those without after undergoing diagenetic change. The bone fractures significantly affected the elemental composition of bone over time, but structural composition initially remained stable. It was also possible to distinguish between perimortem and postmortem fractures using these two analytical techniques due to physicochemical differences. This research shows bone fractures can significantly alter the physicochemical composition of the bone during the postmortem period and have the potential to facilitate more accurate PMI estimations in forensic contexts.