microRNAs as New Biomolecular Markers to Estimate Time since Death: A Systematic Review.

Estimating the post-mortem interval is still one of the most complex challenges in forensics. In fact, the main tools currently used are burdened by numerous limitations, which sometimes allow the time of death to be placed only within too large time intervals. In recent years, researchers have tried to identify new tools to try to narrow down the interval within which to place the time of death; among these, the analysis of microRNAs seems to be promising. An evidence-based systematic review of the literature has been conducted to evaluate the state of the art of knowledge, focusing on the potential correlation between miRNA degradation and PMI estimation. The research has been performed using the electronic databases PubMed, Scopus, and WOS. The results allowed us to highlight the usefulness of miRNAs both as markers for PMI estimation and for normalization, especially due to their stability. In fact, some miRNAs remain particularly stable for long periods and in different tissues, while others degrade faster. Furthermore, there are numerous factors capable of influencing the behavior of these molecules, among which the type of tissue, the cause of death, and the circadian rhythm appear to be the most relevant. Despite the promising results of the few articles present in the literature, because of the numerous limitations they are burdened by, further research is still necessary to achieve more solid and shareable results.

Mummification in a forensic context: an observational study of taphonomic changes and the post-mortem interval in an indoor setting.

The objective of this study was to evaluate the presence of mummification in an indoor setting, with an emphasis on the forensic perspective. A dataset of 102 forensic autopsy cases was assessed for distribution of desiccation of skin and soft tissue (i.e., subcutaneous fat and musculature) and for moist decompositional (i.e., putrefactive) changes. Further, possible correlation with the post-mortem interval (PMI) was evaluated, as well as the effects of clothing coverage of the body. The results indicated that yellow to orange parchment-like desiccated skin was found at significantly shorter PMIs than reddish brown to black leathery desiccated skin, even when soft tissue desiccation was included in the comparative analysis. Clothing appeared to have a significant decelerating effect on the extent of desiccation on the legs, but findings in regard to whole body or torso/arms were inconclusive. A large variation in PMIs was evident as regards fully desiccated skin (PMI 18-217 days), indicating difficulties in PMI estimation due to a variable repressive effect on the decompositional process per se in an indoor setting. For the specific case in forensic practice, no definite conclusion can be drawn from the observed desiccation changes to the PMI. One way forward might be creating a systematic and standardized method for describing different desiccation types, as well as other cooccurring decompositional changes and how they relate to the PMI, as a foundation for a future quantification model.

[Estimation of time since death with the postmortem microbiome: a modern view and approaches to solving the problem]. / Opredelenie davnosti nastupleniya smerti s pomoshch'yu posmertnogo mikrobioma: sovremennyi vzglyad i podkhody k resheniyu problemy.

The aim of the review is to summarize and update the data of modern studies devoted to determining the post-mortem interval (PMI) with the use of microorganisms, as well as disclosing prospects for further study in the presented direction. Estimating the time elapsed since death based on the postmortem microbiome has great potential for accurate determination of PMI, but all methods currently used have their limitations. The dynamics of changes in microbial communities due to the influence of many external and internal factors significantly complicates the process of interpreting the results. The change of microbial communities in the human corpse has shown promising results for the assessment of PMI, but to date there is no evidence of the repeatability of such a continuity in various geographic and ecological conditions. The question of conducting new, large-scale studies, taking in all the factors that could affect the posthumous microbiome, is becoming urgent.

Human Bone Proteomes before and after Decomposition: Investigating the Effects of Biological Variation and Taphonomic Alteration on Bone Protein Profiles and the Implications for Forensic Proteomics.

Bone proteomic studies using animal proxies and skeletonized human remains have delivered encouraging results in the search for potential biomarkers for precise and accurate post-mortem interval (PMI) and the age-at-death (AAD) estimation in medico-legal investigations. The development of forensic proteomics for PMI and AAD estimation is in critical need of research on human remains throughout decomposition, as currently the effects of both inter-individual biological differences and taphonomic alteration on the survival of human bone protein profiles are unclear. This study investigated the human bone proteome in four human body donors studied throughout decomposition outdoors. The effects of ageing phenomena (in vivo and post-mortem) and intrinsic and extrinsic variables on the variety and abundancy of the bone proteome were assessed. Results indicate that taphonomic and biological variables play a significant role in the survival of proteins in bone. Our findings suggest that inter-individual and inter-skeletal differences in bone mineral density (BMD) are important variables affecting the survival of proteins. Specific proteins survive better within the mineral matrix due to their mineral-binding properties. The mineral matrix likely also protects these proteins by restricting the movement of decomposer microbes. New potential biomarkers for PMI estimation and AAD estimation were identified. Future development of forensic bone proteomics should include standard measurement of BMD and target a combination of different biomarkers.

Histological quantification of decomposed human livers: a potential aid for estimation of the post-mortem interval?

The objective of this study was to determine if a novel scoring-based model for histological quantification of decomposed human livers could improve the precision of post-mortem interval (PMI) estimation for bodies from an indoor setting. The hepatic decomposition score (HDS) system created consists of five liver scores (HDS markers): cell nuclei and cell structure of hepatocytes, bile ducts, portal triad, and architecture. A total of 236 forensic autopsy cases were divided into a training dataset (n = 158) and a validation dataset (n = 78). All cases were also scored using the total body score (TBS) method. We specified a stochastic relationship between the log-transformed accumulated degree-days (log10ADD) and the taphonomic findings, using a multivariate regression model to compute the likelihood function. Three models were applied, based on (i) five HDS markers, (ii) three partial body scores (head, trunk, limbs), or (iii) a combination of the two. The predicted log10ADD was compared with the true log10ADD for each case. The fitted models performed equally well in the training dataset and the validation dataset. The model comprising both scoring methods had somewhat better precision than either method separately. Our results indicated that the HDS system was statistically robust. Combining the HDS markers with the partial body scores resulted in a better representation of the decomposition process and might improve PMI estimation of decomposed human remains.

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.

Ecological succession of blow flies (Diptera: Calliphoridae) in the decomposition of pig carcasses in Cerrado areas of Brazil.

This research aims to describe the process of ecological succession by associating the decomposition stages of pig carcasses with flies from the Calliphoridae family (Diptera). For this, 6 pig carcasses were exposed in Maranhão's Cerrado, utilizing metal cages with sawdust trays to catch immature specimens and "suspended traps" to capture adults. Adults of the Hemilucilia benoisti Séguy, and Hemilucilia townsendi Shannon species were only associated with the swelling stage. Chrysomya megacephala (Fabricius) was associated with 2 stages (black putrefaction and fermentation). The species Chloroprocta idioidea (Robineau-Desvoidy), Chrysomya albiceps (Wiedemann), Cochliomyia macellaria (Fabricius), and Lucilia eximia (Wiedemann) were associated with 4 of the 5 stages of decomposition, the latter being the only 1 associated with the initial stage. The larvae abandoned the carcasses to pupate from the second stage of decomposition, with L. eximia being the only 1 leaving the carcasses in the swelling stage, and C. albiceps the only 1 associated with both the fermentation and dry stages. Our findings indicate that calliphorid species can help forensic investigators estimine the post-mortem interval of cadavers in situations similar to those detailed in this study. Since there was a link between adult and immature species and certain stages, they can be used as indicators in future forensic investigations.

Post-Mortem Dental Profile as a Powerful Tool in Animal Forensic Investigations-A Review.

Veterinary forensics is becoming more important in our society as a result of the growing demand for investigations related to crimes against animals or investigations of criminal deaths caused by animals. A veterinarian may participate as an expert witness or may be required to give forensic assistance, by providing knowledge of the specialty to establish a complete picture of the involvement of an animal and allowing the Courts to reach a verdict. By applying diverse dental profiling techniques, not only can species, sex, age-at-death, and body size of an animal be estimated, but also data about their geographical origin (provenance) and the post-mortem interval. This review concentrates on the dental techniques that use the characteristics of teeth as a means of identification of freshly deceased and skeletonised animals. Furthermore, this highlights the information that can be extracted about the animal from the post-mortem dental profile.

Quantifying human decomposition in an indoor setting and implications for postmortem interval estimation.

This study's objective is to obtain accuracy and precision in estimating the postmortem interval (PMI) for decomposing human remains discovered in indoor settings. Data were collected prospectively from 140 forensic cases with a known date of death, scored according to the Total Body Score (TBS) scale at the post-mortem examination. In our model setting, it is estimated that, in cases with or without the presence of blowfly larvae, approximately 45% or 66% respectively, of the variance in TBS can be derived from Accumulated Degree-Days (ADD). The precision in estimating ADD/PMI from TBS is, in our setting, moderate to low. However, dividing the cases into defined subgroups suggests the possibility to increase the precision of the model. Our findings also suggest a significant seasonal difference with concomitant influence on TBS in the complete data set, possibly initiated by the presence of insect activity mainly during summer. PMI may be underestimated in cases with presence of desiccation. Likewise, there is a need for evaluating the effect of insect activity, to avoid overestimating the PMI. Our data sample indicates that the scoring method might need to be slightly modified to better reflect indoor decomposition, especially in cases with insect infestations or/and extensive desiccation. When applying TBS in an indoor setting, the model requires distinct inclusion criteria and a defined population.

Conductometric titration to determine total volatile basic nitrogen (TVB-N) for post-mortem interval (PMI).

Precise measurement of cadaver decomposition rate is the basis to accurate post-mortem interval (PMI) estimation. There are many approaches explored in recent years, however, it is still unsolved completely. Total volatile basic nitrogen (TVB-N), which is an important index to predict meat freshness and shelf life in food science, could serve as an indicator for measuring PMI associated decomposition rate of cadavers. The aim of this work was to establish a practical method to determine TVB-N in cadaver soft tissues (mainly skeletal muscle) for measuring decomposition rate. Determination of TVB-N in the simulation and animal experiments was conducted by steam distillation and conductometric titration using Kjeldahl distillation unit and conductivity meter. In simulation, standard concentrations of ammonium were used as TVB analogies, TVB-N contents were determined and the recovery rates of nitrogen were calculated. In animal experiment, TVB-N in skeletal muscle of forty-two rats was determined at different PMIs for 312 h at 24 °C ± 1 °C. The relationship between PMI and TVB-N was investigated also. The method showed high precision with 99%-100% recovery rates. TVB-N in skeletal muscle changed significantly with PMI especially after 24 h, and the data fit well to y = 3.35 E-5x3-2.17 E-2x2+6.13x-85.82 (adj. R2 = 0.985). ECi (initial electrical conductivity in the samples just before titration) had positive linear relationship to final measured TVB-N values, y = 1.98x+16.16 (adj. R2 = 0.985). The overall results demonstrated that the method is accurate, rapid and flexible, which could be expected as a basic technique for measuring decomposition rate in later PMI-estimation researches. Further studies are needed to validate our findings.

Life history data on the fly parasitoids Aleochara nigra Kraatz and A. asiatica Kraatz (Coleoptera: Staphylinidae), and their potential application in forensic entomology.

Knowledge of the developmental time of the immature stages of necrophagous flies has been the main tool for estimating minimum post-mortem intervals (min PMIs) in forensic entomology. Many parasitic insects can alter the development of immature stages of flies and thus affect min PMI estimates. The larvae of most species of Aleochara rove beetles are ectoparasitoids of the pupae of cyclorrhapha flies. Among them, some species that parasitise necrophagous flies may have forensic importance. Two Taiwanese Aleochara species, A. nigra and A. asiatica, which visit carrion sites were studied herein. All five necrophagous (Hemipyrellia ligurriens, Lucilia cuprina, Chrysomya megacephala, C. rufifacies and sarcophagid sp.) and one non-necrophagous fly species (Bactrocera dorsalis) we examined have the potential to be parasitised by these two Aleochara species, but differences among the acceptability and suitability of these hosts to rove beetle species suggested that rove beetles may prefer specific hosts. Each stage of the beetle life history was recorded to estimate developmental durations at six different temperatures. The larval stage together with the pupal stage of both beetle species was longer than the pupal stages of their hosts, implying the possibility of elongating the min PMI estimation. In addition, the host weight and larval duration of these two Aleochara beetles were positively correlated; thus, potential applications can be expected when using parasitised fly pupae in min PMI estimations.