Cardiac SARS-CoV-2 Infection, Involvement of Cytokines in Postmortem Immunohistochemical Study.

In the context of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection, significant attention was given to pulmonary manifestations. However, cardiac involvement is increasingly recognized as a critical factor influencing the prognosis, leading to myocardial damage, heart failure, acute coronary syndromes, potentially lethal arrhythmic events, and sudden cardiac death. Despite these findings, there is a lack of studies detailing the necroscopic, macroscopic, and microscopic cardiac changes associated with SARS-CoV-2. This study aimed to investigate the presence of SARS-CoV-2 viral proteins in cardiac tissue using immunohistochemical techniques to assess viral tropism. The analysis of cardiac tissue samples from deceased subjects, in different stages of conservation, confirmed to be positive for SARS-CoV-2 via reverse transcriptase-polymerase chain reaction (RT-PCR), showed immunopositivity for the SARS-CoV-2-NP viral antigen in 33% of cases. Notably, the presence of leukocyte infiltrates sufficient for diagnosing lymphocytic myocarditis was not observed. The central proinflammatory cytokines involved in the pathogenetic mechanism of coronavirus disease 19 (COVID-19) were researched using the immunohistochemical method. A significant increase in cytokine expression was detected, indicating myocardial involvement and dysfunction during SARS-CoV-2 infection. These findings suggest that the immunohistochemical detection of SARS-CoV-2 viral antigens and inflammatory cytokine expression in cardiac tissue could be crucial for a proper forensic assessment of the cause of death, even in sudden cardiac death.

Depressed fractures of the skull due to direct kick and the contribution of 3D CT reconstruction.

Depressed skull fractures occur when broken bones displace inward, meaning that a portion of the outer table of the fracture line lies below the normal anatomical position of the inner table. They typically result from force trauma, when the skull is struck by an object with a moderately large amount of kinetic energy but a small surface area, or when an object with a large amount of kinetic energy impacts only a small area of the skull. In the present case, a depressed fracture of the frontal bone was detected at the autopsy of a 52-year-old man who, according to the belated confession of the assailant, was kicked in the head. The assailant was wearing sneakers. Could such a fracture be caused "just" by a kick? In this case it was possible due to an extraordinarily thin cranial vault (0.2 cm frontal, 0.3 cm occipital), which allowed the fractures to occur from a kinetic force that might not have been sufficient with a normal cranial vault thickness. An important role in the forensic analysis of the case was played by the 3D CT reconstruction.

Exploring the role of the human microbiome in forensic identification: opportunities and challenges.

Forensic microbiology is rapidly emerging as a novel tool for human identification. The human microbiome, comprising diverse microbial communities including fungi, bacteria, protozoa, and viruses, is unique to each individual, offering a new dimension to forensic investigations. While traditional identification methods primarily rely on DNA profiling and fingerprint analysis, they face limitations when complete DNA or fingerprints profiles are unattainable or degraded. In this context, the microbial signatures of the human skin microbiome present a promising alternative due to their resilience to environmental stresses and individual-specific composition. This review explores the potential of microbiome analysis in forensic human identification, evaluating its applications, advantages, limitations, and future prospects. The uniqueness of an individual's microbial community, particularly the skin microbiota, can provide distinctive biological markers for identification purposes, while technological advancements like 16 S rRNA sequencing and metagenomic shotgun sequencing are enhancing the specificity of microbial identification, enabling detailed analysis of these complex ecological communities. Despite these promising findings, current research has not yet achieved a level of identification probability that could establish microbial analysis as a stand-alone evidence tool. Therefore, it is presently considered ancillary to traditional methods, contributing to a more comprehensive biological profile of individuals.

Intentional child and adolescent homicides in Milan (Italy): A 30-year interdisciplinary study.

This study aims to discuss the forensic and criminological implications of child homicides in the territory of Milan, Italy. The authors present a retrospective study on all the cases of child and adolescent homicides, that were observed at the Institute of Legal Medicine of Milan (Italy) in the last 30 years (from January 1991 to December 2020). A total of 46 child homicides were collected, focusing on the sociological features, by highlighting peculiar cases, risk factors, potential changing social trends, and comparing our cases with the current literature. The analysis revealed a statistically significant relationship (p < 0.05) for male adolescents and indicated that adolescent homicides were more frequently perpetrated in extrafamilial contexts. In contrast, neonaticides and infanticides were mainly committed at home. Furthermore, the Fisher's Exact test revealed that child murders were mainly committed by immigrants in the Milan district after 2005 (p < 0.05). The two main causes of death were due to sharp and firearm injuries. While the relationship between homicide clusters and homicides committed by sharp objects was not significant, adolescent homicides were mainly committed using firearms (p < 0.05). The present study may help to identify risk factors for homicides against child and adolescent. Consequently, policies that identify, prevent, and minimize this extreme violence should be designed to interrupt the vicious circle of such dreadful murderous events. Child and adolescent homicides deserve additional focus and better education for healthcare professionals and further research should be carried out to develop therapeutic and caring strategies.

Pathological Findings in Hanging: Is the Traditional Knowledge Correct?

BACKGROUND: In forensic pathology, asphyxia presents a problem as frequently as it is thorny and challenging. Some knowledge in forensic pathology is still considered to be traditionally acquired and is not critically assessed with modern statistical or technical tools. In this study, we seek to examine the injuries that are considered to be typical of cases of hanging (neck lesions, haemorrhages, and pleural and epicardial petechiae). METHODS: We evaluated whether there was, indeed, a statistically significant association between these injuries and deaths from hanging as compared to non-hanging deaths. We collected 399 cases (32 deaths by hanging and 367 cases of non-hangings), built contingency tables and performed chi-square tests for each variable (lesion) examined; we also analysed this association in various subgroups of the sample (according to sex, age and weight ranges). RESULTS: Our results did not deviate from the expected outcome based on traditional knowledge, although they do provide a more detailed demonstration and clarification of traditional knowledge regarding this topic. CONCLUSIONS: These findings provide valuable insights for future discussion, examination and deepening of knowledge that is traditionally accepted but often undersupported in the literature.

Why identification matters: an explorative study on six cases of family reunification.

The escalating phenomenon of migration, accompanied by a disturbing surge in associated tragedies, has persistently violated internationally protected human rights. Absence of physical evidence, namely the presence of adequately identified corpses, may impede the full enjoyment of human rights and-in some cases-the course of justice as it obstructs the initiation of legal proceedings against individuals implicated in causing such catastrophes. It also presents administrative obstacles, as death certificates are indispensable in legitimizing statuses like orphanhood and widowhood. Family reunification, particularly for orphans, plays a significant role for those attempting to reconnect with their relatives all over the world. Likewise, for mothers, the acknowledgment of their marital status or widowhood can be a pathway to regain their marginalized right to social life. To elucidate this issue, we analyzed six representative cases from the tragic October 3, 2013, shipwreck near the Italian island of Lampedusa, where 366 individuals were retrieved dead from the sea. These cases underscore the practical challenges involved, highlighting the compelling need for continued efforts to ensure that this burdensome problem transcends from being a mere ethical, moral, and legal discourse. Although considerable progresses, these cases also reveal that substantial work still lies ahead. There is a pressing need for improved mechanisms to certify kinship ties, which are often the limiting factor in many reunifications, and can hinder the granting of custody to children. The severity and far-reaching implications of this problem necessitate thoughtful attention and action, especially considering the ongoing escalation in migration and related fatalities.

Plastic bag suffocation and codeine overdose: An unusual case of complex suicide and review of the literature.

The term "planned complex suicide" refers to cases where two or more suicide methods are applied simultaneously. Plastic bag suffocation is a suicidal method commonly combined with self-poisoning by inhalation of volatile compounds or pharmacological substances at poisoning dosage. A 67-year-old woman was found dead on the couch. The head was wrapped in a plastic bag knotted at the front of the neck. No signs of struggle were present on the scene. In the fireplace, there were two blisters of acetaminophen/codeine phosphate tablets. Postmortem examination showed congestion of the face and the neck and pulmonary edema with patchy emphysema. There was no evidence of physical duress. Toxic levels of codeine were detected in the blood. The death was related to a complex suicide involving codeine overdose and asphyxia by plastic bag suffocation. The hypoxic/cardioinhibitory effects of plastic bag suffocation combined to codeine-induced deep reduction of respiratory rate, making the suicide hypothesis the more likely scenario. In order to assess what elements contribute to postmortem diagnosis in complex suicides with plastic bag suffocation, a review of the forensic literature published between 2002 and 2022 was performed. This article emphasizes the complexity of diagnosing deaths involving plastic bag suffocation in suicides, particularly when combined with other methods like inert gas inhalation or drug intake. It underscores the importance of comprehensive toxicological analyses, careful scene examination, and histological studies, not forgetting a thorough comparative analysis of the existing literature, to accurately determine the cause and manner of death in such cases.

Skin surface debris as an archive of environmental traces: an investigation through the naked eye, episcopic microscope, ED-XRF, and SEM-EDX.

Traces from bodies can be of various nature, for example of biological or inorganic origin. Some of these historically have received more consideration than others in forensic practice. Samplings of gunshot residues or biological fluid traces are commonly standardized, whereas macroscopically invisible environmental traces are usually ignored. This paper simulated the interaction between a cadaver and a crime scene by placing skin samples on the ground of five different workplaces and inside the trunk of a car. Traces on samples were then investigated through different approaches: the naked eye, episcopic microscope, Scanning Electron Microscopy (SEM) with Energy-dispersive X-ray spectroscopy (EDX) and Energy Dispersive X-Ray Fluorescence (ED-XRF). The purpose is to provide the forensic scientist with the awareness of the value of debris on skin and then to highlight implications for forensic investigations. Results demonstrated that even naked eye observation can reveal useful trace materials, for defining the possible surrounding environment. As a next step, the episcopic microscope can increase the number of visible particulates and their analysis. In parallel, the ED-XRF spectroscopy can be useful to add a first chemical composition to the morphological data. Finally, the SEM-EDX analysis on small samples can provide the greatest morphological detail and the most complete chemical analysis, although limited, like the previous technique, to inorganic matrices. The analysis of debris on the skin, even with the difficulties due to the presence of contaminants, can provide information on the environments involved in criminal events that can add to the investigation framework.

Transcending time: the forensic anthropological case study of three unidentified transgender women in Italy in the early 1990s.

In forensic anthropology, the estimation of skeletal sex results in a dichotomous outcome: male or female. This poses challenges when dealing with transgender individuals. Italy has the highest rate per capita of transgender murders, making their identification a compelling issue. This study presents three cases of unidentified skeletal remains of transgender women (MtF) that underwent autopsies in the 1990s at the Institute of Legal Medicine of Milan. The transgender status of the individuals was determined from the autopsy records which indicated the co-existence of breast implants and male genitalia. The biological profile was created by estimating sex, age, ancestry, and stature, and the bones were investigated looking for evidence of Facial Feminization Surgery (FFS). Anthropological estimations revealed that the skeletons belong to male, adults, European individuals, between 164 and 184 cm. Fragmented gelatinous masses were found in association with two skeletons, which were interpreted as remnants of breast implants. In two cases, signs of remodeling of the cortical surface were observed on the zygomatic bones, although the bone marks observed here were not specific enough to link them to FFS. Despite some limitations, this study highlights the need for greater awareness among practitioners about this limitedly addressed issue, advocating for a more inclusive forensic anthropology that strives to improve methods and interpretation of evidence for the identification of transgender individuals.

Toxicological analysis in victims of sexual and domestic violence: A retrospective study of a 3-year period (2018-2020).

This manuscript presents an epidemiological investigation carried out on abuse victims who accessed the Sexual and Domestic Violence Service (SVS&D) of IRCCS Ca' Granda in Milan, Italy. The focal point of this research was the detection of alcohol, prescription medications, and illicit substances in victims who solicited help from the SVS&D center between 2018 and 2020. Over this three-year span, biological samples of blood and urine were procured from 207 victims, out of a patient pool of 2470. All collected samples were analyzed via High Performance Liquid Chromatography - Tandem Mass Spectrometry (HPLC-MS/MS) and Gas Chromatography - Mass Spectrometry (GC-MS). Toxicological examination results demonstrated that 43% of the cases tested positive for substances in 2018, 39% in 2019 and 60% of the cases in 2020. Overall, 45% of the victims tested resulted positive to some substance over a 3-year period, equivalent to 3.6% of the overall cases (2470 victims). Substances of toxicological interest were detected in 104 samples (out of 377, corresponding to 27.6%) belonging to 94 patients. The most detected classes of drugs were stimulants, antidepressants, benzodiazepines and antipsychotics. Moreover, BAC (Blood Alcohol Concentration) indicated positivity in 25 cases (out of 184 cases analyzed - 14% of positive cases). Based on this study's findings, we recommend broadening the range of substances evaluated in drug-facilitated sexual assaults and establishing standardized protocols for both national and international implementation. Implementing procedures would significantly enhance forensic support provided to victims of abuse seeking healthcare services post-incident.

An unusual case of a triple suicide pact at the time of the COVID-19 pandemic.

In December 2020, the World Health Organization (WHO) declared SARS-CoV2 a global pandemic. Home confinement, low social contacts, and fear of virus transmission played a major role as risk factors for suicides during the following period. Suicide pacts, in particular, showed a different pattern. A rare case of a triple suicide pact among members of the same family nucleus is presented. The victims were an elderly, severely ill woman and her adult children (a son and daughter), linked by a morbid relationship. The last time the family was seen alive was 40 days before the discovery. All corpses presented decompositional changes. After a full autopsy, the cause of death was determined to be a lethal intake of morphine for the mother and acute blood loss due to self-stabbing at the neck for the siblings. The younger woman was under the effects of a large amount of heparin. Toxicological analysis was positive for opioids and alcohol in both siblings. Suicide pacts have rarely been described during the COVID-19 pandemic. In the few cases reported, the victims were more often relatives than people in a romantic relationship. The involvement of three people is unusual, as is the use of different suicide methods among the victims. In the presented case, the elderly mother's imminent death from terminal cancer, her concern over dying in a nondomestic environment, and the siblings' fear of being alone likely led to the conception of the suicide pact. Social isolation and economic difficulties also played a contributing role.

Estimation of Late Postmortem Interval: Where Do We Stand? A Literature Review.

Estimating time since death can be challenging for forensic experts, and is one of the most challenging activities concerning the forensic world. Various methods have been assessed to calculate the postmortem interval on dead bodies in different stages of decomposition and are currently widely used. Nowadays, the only well-recognized dating technique is carbon-14 radioisotope measurement, whereas other methods have been tested throughout the years involving different disciplines with different and sometimes not univocal results. Today, there is no precise and secure method to precisely determine time since death, and late postmortem interval estimation remains one of the most debated topics in forensic pathology. Many proposed methods have shown promising results, and it is desirable that with further studies some of them might become acknowledged techniques to resolve such a difficult and important challenge. The present review aims at presenting studies about the different techniques that have been tested in order to find a valuable method for estimating time since death for skeletal remains. By providing a comprehensive overview, the purpose of this work is to offer readers new perspectives on postmortem interval estimation and to improve current practice in the management of skeletal remains and decomposed bodies.

Review on forensic importance of myiasis: Focus on medicolegal issues on post-mortem interval estimation and neglect evaluation.

Myiasis is the infestation of live vertebrates by dipterous larvae that feed on living or necrotic tissues, liquid body substances or ingested food and develop in or on the vertebrate body. In both animals and humans, myiasis plays a fundamental role in forensic practice because of its implications in the evaluation of cases of neglect and in the mPMI (minimum post-mortem interval) estimation. The present study aims to provide a review of forensic issues related to myiasis. A retrospective analysis was conducted by exploring major electronic literature databases. Methodological evaluation of each study was performed according to the PRISMA (Preferred Reporting Items for Systematic Review and Meta-Analyses) standards. Sixty-one papers were included in this review. Entomology shows great potentiality in forensic investigations, mostly because of its contributory role in mPMI estimation. Such evaluation, however, requires a careful consideration of the possibility of an ante-mortem myiasis infestation, which could complicate the correct assessment of the mPMI. Hence, forensic pathologists should pay attention to the entomological fauna on a corpse, in both fresh and decomposed bodies. Moreover, in cases with poor nutritional or hygienic conditions, myiasis should be carefully evaluated as an indicator of neglect.

Mass spectrometry-based proteomic strategy for ecchymotic skin examination in forensic pathology.

Mass spectrometry (MS)-based proteomics has recently attracted the attention from forensic pathologists. This work is the first report of the development of a shotgun bottom-up proteomic approach based on rapid protein extraction and nano-liquid chromatography/high-resolution mass spectrometry applied to full-thickness human skin for the differential analysis of normal and ecchymotic tissues to identify new biomarkers for bruise characterization and dating. We identified around 2000 proteins from each pooled extract. The method showed excellent precision on independent replicates, with Pearson correlation coefficients always higher than 95%. Glycophorin A, a known biomarker of vital wounds from immunochemical studies, was identified only in ecchymotic tissues, as confirmed by Western blotting analysis. This finding suggests that this protein can be used as a MS-detectable biomarker of wound vitality. By focusing on skin samples from individuals with known wound dating, besides Glycophorin A, other proteins differentially expressed in ecchymotic samples and dependant on wound age were identified, although further analysis on larger datasets are needed to validate these findings. This study paves the way for an in-depth investigation of the potential of MS-based techniques for wound examination in forensic pathology, overcoming the limitations of immunochemical assays.

The "mystery" of the train station: The importance of a forensic approach to a rare case of survival after a train-pedestrian collision.

In the early morning a 28-year-old man was found lying on the tracks of a railway station with head injuries and fractures of the cervical spine resulting in permanent quadriplegia. He was in a club about 1 km away until about 2 h earlier and did not have any recollection of what could have happened. Was he the victim of an assault, did he fall down or was he hit by a passing train? The solution to this "mystery" came from a forensic evaluation that included the forensic branches of pathology, chemistry, merceology and genetics as well as the scene evaluation. Through these different steps, the role of a railway collision in determining the injuries was ascertained and a possible dynamic was postulated. The presented case is an expression of the importance of the different forensic disciplines and the difficulties the forensic pathologist encounters when analysing such peculiar and rare cases.

Physical assessment of tortures: Interobserver analysis of medico-legal degrees of consistency based on the Istanbul protocol.

The Istanbul Protocol section on medicolegal evaluation delineates the guidelines for the correct management of the physical examination and the methods of assigning the degrees of consistency. Considering that most cases exhibit highly heterogeneous lesions, the examiner is forced to rely on his own experience, and their evaluation may be very subjective. The purpose of this work is to understand how subjective such an evaluation may be and whether the "experience" factor, interpreted as years dedicated to this profession and the number of cases evaluated, is statistically significant. To this end, a survey containing eleven cases of pre-evaluated asylum seekers was sent to thirty Italian clinical forensic practitioners. The participants were invited to assign a degree of consistency to each case according to the Istanbul Protocol guidelines, besides answering a few questions regarding their professional record. The doctors were divided into groups based on the number of cases evaluated and the experience collected expressed in years, and then interobserver analysis was performed. Results showed that the Fleiss' Kappa coefficient acquired significant values when attention was turned to the sub-samples composed of more experienced participants. Therefore, the introduction of appropriately trained health professionals - "experts in migrations and torture" - could lower the risks of misinterpretation and make the assessment as reproducible as possible.