Age assessment in unaccompanied minors: assessing uniformity of protocols across Europe.

Age assessment of migrants is crucial, particularly for unaccompanied foreign minors, a population facing legal, social, and humanitarian challenges. Despite existing guidelines, there is no unified protocol in Europe for age assessment.The Forensic Anthropology Society of Europe (FASE) conducted a comprehensive questionnaire to understand age estimation practices in Europe. The questionnaire had sections focusing on the professional background of respondents, annual assessment numbers, requesting parties and reasons, types of examinations conducted (e.g., physical, radiological), followed protocols, age estimation methods, and questions on how age estimates are reported.The questionnaire's findings reveal extensive engagement of the forensic community in age assessment in the living, emphasizing multidisciplinary approaches. However, there seems to be an incomplete appreciation of AGFAD guidelines. Commonalities exist in examination methodologies and imaging tests. However, discrepancies emerged among respondents regarding sexual maturity assessment and reporting assessment results. Given the increasing importance of age assessment, especially for migrant child protection, the study stresses the need for a unified protocol across European countries. This can only be achieved if EU Member States wholeheartedly embrace the fundamental principles outlined in EU Directives and conduct medical age assessments aligned with recognized standards such as the AGFAD guidelines.

Atypical Postmortem "Incaprettamento" After an Honor Homicide.

ABSTRACT: Honor killing is a murder carried out to protect the family's reputation by actions that are perceived as shameful. Usually, honor killing occurs to preserve family's honor, which has been violated by adulteries, premarital pregnancies, or inappropriate relationships. In Southern Italy, the culture of honor is quite strong especially among the local criminal organizations. Honor crimes by Italian Mafia are intended to have an admonitory significance, and they can be carried out following a macabre ritual of "incaprettamento." The aim is to humiliate the victim treating him like an animal, a bonded young goat where the victim is secured by a rope passing behind the back from the upper to lower extremities. In this case study, the honor homicide was perpetrated against the lover of a butcher's wife by sharp weapon, followed by a postmortem atypical incaprettamento of the corpse. The wrists and ankles were tied together by ropes, and the extremities were fixed to the abdomen with a string. The body was also wrapped in a plastic tape after penile amputation. According to killer's confession, the mutilation and the incaprettamento were expressions of revenge and contempt for the victim to restore the honor lost.

COVID-19: detection methods in post-mortem samples.

COVID-19 identification is routinely performed on fresh samples, such as nasopharyngeal and oropharyngeal swabs, even if, the detection of the virus in formalin-fixed paraffin-embedded (FFPE) autopsy tissues could help to underlie mechanisms of the pathogenesis that are not well understood.The gold standard for COVID-19 detection in FFPE samples remains the qRT-PCR as in swab samples, contextually other methods have been developed, including immunohistochemistry (IHC), and in situ hybridization (ISH). In this manuscript, we summarize the main data regarding the methods of COVID-19 detection in pulmonary and extra-pulmonary post-mortem samples, and especially the sensitivity and specificity of these assays will be discussed.

Colorectal Cancer: 35 Cases in Asbestos-Exposed Workers.

BACKGROUND: Asbestos is considered one of the major global work-related carcinogens. Some studies suggest a potential causal relationship between asbestos exposure and colorectal cancer (CRC). However, the role of asbestos in CRC carcinogenesis is still controversial. METHODS: 35 claims of occupational CRC among asbestos-exposed workers were reviewed. All claims were rejected by the Italian National Institute for Insurance against Accidents (INAIL) due to the "lack of scientific evidence in the causality" between asbestos exposure and CRC; causality was finally assessed in civil trials. All cases were categorized by age, gender, industry type, task, exposure and latency periods, anatomical location, and histopathological characteristics of CRC and concomitant respiratory diseases. RESULTS: Most workers were males aged 60 years or more and employed in occupational activities with extensive use of asbestos for over 20 years. In 31 out of 35 cases, CRC was diagnosed over 39 years after employment. Right-sided colic adenocarcinoma was diagnosed in nine cases; rectum was involved in eight cases. Respiratory comorbidities were observed in 22 workers. CONCLUSIONS: Our study provides some interesting points in the assessment of the causal relationship between asbestos exposure and CRC.

Choking in patients with neurological disorders and role of drug-induced dysphagia.

Choking is a type of asphyxia due to the internal obstruction of airways by foreign material, quite always food. Most choking deaths are accidental and occur so quickly that may mimic a heart attack. This scenario is also known as "Cafè coronary syndrome" or "bolus death". At autopsy, pieces of under-chewed food are commonly lodged at, above, or in the tracheal space adjacent to the laryngeal inlet. In these fatal events mechanisms other than asphyxia can be also involved, such as a vasovagal episode (by stimulation of the autonomic nerve plexus of the laryngeal inlet) or swallowing impairment. Swallowing is a complex, semi-automatic process that can be affected by various disorders (i.e. dementia, Parkinson, neurological injuries, etc.), senility and external factors causing oropharyngeal dysphagia (OD). Among these factors, several drugs have been also associated with impaired swallowing, including drugs acting on the CNS like antipsychotics and antiepileptics. Three cases of witnessed bolus deaths are reported. All victims were affected by neurological defects and took medications acting on CNS. In all cases, at autopsy pieces of food were found distributed from the laryngeal inlet along the main axis of the trachea up to the large bronchi over the bifurcation. Additional autopsy findings were represented by facial congestion and cyanosis, subepicardial petechiae and pulmonary emphysema.

Matricide and psychiatric evaluation: An update.

Matricide is an infrequent crime that has often raised to the suspicion that the offender could suffer from a pathological mental status. Although it is hard to establish a relationship between mental disorders and specific forms of homicide, several studies suggest that matricide offender frequently suffers from schizophrenia or other psychotic disorders. Aim of the study was to review the literature in order to explore epidemiologic and psychiatric characteristics of matricide offenders with a focus on the cause of death and crime circumstances. According to the PRISMA statement, 16 out of 225 studies were included. Main findings were the following: 80 victims were reported in total, killed by 81 offenders. In one case two brothers were responsible for the matricide. 81.5% of the offenders were young males. The most represented psychiatric pattern of the offenders was schizophrenia and psychotic disorders (43.2%). 6.2% of victims had also psychiatric disorders. Sharp force injuries were the first cause of death (55%), followed by blunt trauma (15%) and asphyxia (15%). In 12% of cases overkilling was also reported. 13.6% of offenders were considered not guilty for reason of insanity while 25.9% of the offenders had diminished criminal responsibility. A case study of a young homicide offender suffering from personality disorder is reported. In this case the victim also suffered from psychiatric disorders and an overkilling occurred as she was stabbed multiple times post-mortem. In a second case of matricide, the victim was dismembered and the human remains were concealed in several plastic bags.

Hypoplastic Left Heart Syndrome: About a Postnatal Death.

BACKGROUND: Hypoplastic left heart syndrome (HLHS) is a congenital heart disease that is associated with high mortality rates in the early neonatal period and during surgical treatments. This is mainly due to missed prenatal diagnosis, delayed diagnostic suspicion, and consequent unsuccessful therapeutic intervention. CASE REPORT: twenty-six hours after birth, a female newborn died of severe respiratory failure. No cardiac abnormalities and no genetic diseases had been evidenced or documented during intrauterine life. The case became of medico-legal concern for the assessment of alleged medical malpractice. Therefore, a forensic autopsy was performed. RESULTS: the macroscopic study of the heart revealed the hypoplasia of the left cardiac cavities with the left ventricle (LV) reduced to a slot and a right ventricular cavity that simulated the presence of a single and unique ventricular chamber. The predominance of the left heart was evident. CONCLUSIONS: HLHS is a rare condition that is incompatible with life, with very high mortality from cardiorespiratory insufficiency that occurs soon after birth. The prompt diagnosis of HLHS during pregnancy is crucial in managing the disease with surgery.

Evaluation of Muscle Proteins for Estimating the Post-Mortem Interval in Veterinary Forensic Pathology.

Postmortem cadaveric changes are commonly used to estimate the postmortem interval (PMI) in humans and animals. However, these modifications have been poorly investigated in animals of interest to veterinary forensic pathology. The aim of this study was to investigate the potential use of muscle proteins (desmin and dystrophin) as biomarkers for estimating the PMI in dogs. For this study, 10 dead adult dogs were evaluated for 4 days in a temperature-controlled room at 19 ± 1 °C. For each animal, at 3, 24, 48, 72, and 96 h after death, a 1 × 1 × 1 cm cube of muscle tissue was removed from the vastus lateralis and triceps brachii. Protein expression levels were analyzed by immunohistochemical examination and immunoblot analysis. The obtained results showed rapid dystrophin degradation, with complete disappearance at 72 h after death. In contrast, desmin-positive fibers and desmin protein bands detected by immunoblot were observed on all 4 days of observation. Our findings suggest the potential use of muscle proteins as biomarkers for estimating the PMI in dogs.

Gunshot injuries in Campania (Italy): A retrospective thirty-years (1981-2011) study.

From 1981 to 2011, 200 firearm deaths were autopsied in Naples, at the Institute of Legal Medicine of the University of Campania "Luigi Vanvitelli". 188 cases were homicides and most of them (116) were related to the local organized crime. The majority of victims were young Italian males in the 20-39 age fired in outdoor environments. The reason for choosing outdoor environments can be related to the opportunity for the killer run from the crime scene immediately after the murder. Only 11 of the bodies autopsied were suicide victims, mostly older individuals, over 50 years -old with history of mental illness. All the suicides occurred in indoor environments in order to protect their domestic intimacy. Only two female victims were accounted in this historical series which is quite impressive if compared to recent phenomenon of feminicides occurring mostly in domestic environments. A total of 772 entry wounds were observed: 658 fired from single-charge handguns and 114 from multiple-charge firearms. 9x21 pistol cartridge was the most common ammo used, followed by the 7.65 Parabellum. The head was the most common anatomical site injured (in 81.8% of the suicides cases and in 68.6% of homicides). Most of the victims of homicide died before arriving at an Emergency Service. Only a minority of victims lived from few hours up to less than a week, after being shot, and very few victims remained alive up to a couple of months.

Accidental Injury or "Shaken Elderly Syndrome"? Insights from a Case Report.

Subdural haemorrhage (SDH) as result of a traumatic brain injury (TBI) is a common cause of death in cases of fatal physical abuse. Since intracranial bleeding is a common finding in elderly due to age-related intracranial changes or increasing prevalence of anticoagulant medication, differential diagnosis between inflicted and non-inflicted head injury is challenging. A case of an elderly woman's death caused by TBI is reported. Autopsy showed multiple polychromatic bruises and a frontoparietal hematoma with bilateral subacute SDH. History excluded paraphysiological or pathological non-traumatic conditions that could justify SDH, while iatrogenic factors only played a contributory role. Since polychromatic bruises distributed on the face, the upper extremities and the chest were consistent with forceful grasping/gripping or repeated blows and SDH can form in absence of impact or by mild/minor blows, SDH was considered the result of repeated physical abuses. Differential diagnosis between traumatic and non-traumatic SDH is still challenging for forensic pathologists. As largely accepted in the pediatric population and occasionally described also in adults, however, violent shaking should be also considered as a possible mechanism of SDH-especially in elderly who do not have any sign of impact to the head.

Chronic Encapsulated Intracerebral Hematoma as an Occasional Finding in Sudden Cardiac Death.

Chronic encapsulated intracerebral hematoma (CEIH) is a rare solid mass characterized by the presence of a fibrotic capsule that can present a variety of signs and symptoms due to the mass effect and hydrocephalus. It may be caused by post-traumatic or spontaneous bleeding as related to an adjacent aneurysm, angiomas or neoplasms. Differential diagnosis must be applied in order for it to mimic neoplasm or a vascular malformation. Several cases of CEIH have been reported but only a few of them have an intraventricular localization. A forensic autopsy of a 50-year-old male who died suddenly while driving is discussed. Gross analysis, histology and toxicology were performed and a CEIH of the right lateral ventricle was found in a case of acute coronary death.

Helium Suicide, a Rapid and Painless Asphyxia: Toxicological Findings.

Suicide by helium inhalation has become increasingly common in the last few decades in Europe and the US because it produces a quick and painless death. Inhaled-gas suicides can easily be assessed through death scene investigation and autopsy. However, helium is a colorless and odorless inert gas that unfortunately cannot be detected using standard toxicological analysis. A successful gas analysis was performed following the suicide of a 17-year-old female. For the detection of helium, central/peripheral blood samples and gaseous samples from the esophagus, stomach, and upper and lower respiratory airways (from the trachea and the primary left and right bronchia) were collected with a gastight syringe, ensuring minimal dilution. Qualitative analyses were positive in all gaseous samples. Quantitative analyses were performed using a special gas-inlet system with a vacuum by which the sample can be transferred to a mass spectrometer, reducing the risk of contamination. Helium concentrations were 20.16% from the trachea, 12.33% from the right lung, and 1.5% from the stomach. Based on the high levels of helium, the cause and manner of death were assessed as asphyxia suicide by inhalation of helium. Therefore, toxicological analyses should always be applied in order to gain evidence of inhaled gas in gaseous samples.

Toxicological Analysis of Illicit Drugs Seized in Naples (Italy) and First Detection of Synthetic Cannabinoids during COVID-19 Pandemic.

The COVID-19 pandemic has consistently raised the number of drug seizures, in United States as in Europe. The COVID-19 pandemic has also changed the typology of seizures from "more traditional" drugs to New Psychoactive Substances (NPSs), depending on geographical area. In Europe, the most frequent NPSs are synthetic cannabinoids (SCs) and cathinones, nonetheless synthetic opioids and phenethylamines are widely used. The aim of the study is the detection of NPS and other substances of abuse available in the black market, by quali/quantitative methods in LC-MS/MS and GC-MS. From 2018 to 2021, 268 seizures occurred in total and were analyzed by the Forensic Toxicology Laboratory (FTL) of Naples (Italy). The distribution of analyzed seizures over the years is the following: 53 in 2018, 61 in 2019, 89 in 2020 and 65 during the first semester of 2021. Cannabis was the most detected drug both in hashish and marijuana seizures, followed by cocaine > heroine > prescribed drugs > ketamine−amphetamine MDMA. No NPSs were seized until June 2021, when NPSs were found in two different seizures: Case #1 showed a bar of Cannabis resin containing a low level of Δ9THC = 0.57% associated to SC AB-FUBINACA in 4.5%; Case #2 showed a vegetal resinous substance (Δ9THC = 0.27%) with SC 5F-APINACA (4.1%) associated with methadone (1.0%). The detection of NPSs is alarming evidence that can lead to an increase in the risk of overdose or other negative and unpredictable consequences, such as violent or self-harming behavior in unaware users of cannabis derivatives considered of "natural" origin.

Relationship between COVID-19 Mortality, Hospital Beds, and Primary Care by Italian Regions: A Lesson for the Future.

One of the characteristics of the SARS-CoV-2 infection in Italy is the significant regional difference in terms of lethality and mortality. These geographical variances were clear in the first wave and confirmed in the second one as well. The study aimed to analyze the correlation between regional differences in COVID-19 mortality and different regional care models, by retrospectively analyzing the association between the Italian COVID-19 deaths and the number of hospital beds, long-term care facilities, general practitioners (GPs), and the health expenditure per capita. The period considered was from 1 March 2020 to 1 March 2021. The number of hospital beds (p < 0.0001) and the number of GPs (p = 0.0094) significantly predicted the COVID-19 death rate. The Italian regions with a higher number of hospital beds and a lower number of GPs showed a higher number of deaths. Multivariate analyses confirmed the results. The Italian regions with a higher amount of centralized healthcare, as represented by the number of hospital beds, experienced a higher number of deaths, while the regions with greater community support, as exemplified by the number of the GPs, faced higher survival. These results suggest the need for a change in the current healthcare system organization.

PD-L1 Overexpression in the Lungs of Subjects Who Died from COVID-19: Are We on the Way to Understanding the Immune System Exhaustion Induced by SARS-CoV-2?

Knowledge of the pathogenic mechanisms of severe acute respiratory syndrome-associated coronavirus 2 (SARS-CoV-2) is certainly a priority for the scientific community. Two main elements are involved in the biology of the most severe forms of coronavirus disease 2019 (COVID-19): the direct cytopathic effect of the virus against the host tissues, and a dysfunction of the immune system, characterized by the exhaustion of T lymphocytes. The exhaustion of T cells in COVID-19 is poorly understand, but some data could suggest a possible role of PD-1/PD-L1 axis. The aim of this study was to evaluate the possible role of PD-L1 expression in the pulmonary tissue in subjects affected by COVID-19. The presence of SARS-CoV-2 in the pulmonary tissue, and its exact location, was indagated by in situ hybridization; the expression of PD-L1 and CD8 in the same tissue was indagated by immunohistochemistry. Overall, PD-L1 resulted diffusely expressed in 70% of the cases, and an intense expression was observed in 43.5% of cases. Diffuse and intense presence of SARS-CoV-2 by in situ hybridization significantly correlated with an intense PD-L1 expression, and with expression of PD-L1 by pneumocytes. PD-L1 is overexpressed in the pulmonary tissue of subjects died from COVID-19, and mainly in subjects with a high viral load. These data suggest a possible role of PD-L1 in the immune system exhaustion at the basis of the severe forms of the disease.

Suicide by Pesticide (Phorate) Ingestion: Case Report and Review of Literature.

It has been estimated that approximately one in seven of all global suicides is due to pesticide self-poisoning, mostly in rural areas of developing countries. Organophosphorus (OP) compounds are a group of pesticides exerting their toxicological effects through non-reversible inhibition of the enzyme acetylcholinesterase (AChE). Among these compounds, phorate (thimet) is one of the most dangerous compounds, the use of which is restricted in many countries. A case of intentional suicide after phorate ingestion in a 24-year-old Bengali male is described. This is the second case of suicidal ingestion of phorate reported in the forensic literature, and the first presenting complete toxicological findings.

MALDI-TOF Mass Spectrometry Analysis and Human Post-Mortem Microbial Community: A Pilot Study.

Introduction: The human post-mortem microbiome (HPM) plays a major role in the decomposition process. Successional changes in post-mortem bacterial communities have been recently demonstrated using high throughput metagenomic sequencing techniques, showing great potential as a post-mortem interval (PMI) predictor. The aim of this study is to verify the application of the mass spectrometry technique, better known as MALDI-TOF MS (matrix-assisted laser desorption/ionization time-of-flight mass spectrometry), as a cheap and quick method for microbe taxonomic identification and for studying the PM microbiome. Methods: The study was carried out on 18 human bodies, ranging from 4 months to 82 years old and with a PMI range from 24 h up to 15 days. The storage time interval in the coolers was included in the final PMI estimates. Using the PMI, the sample study was divided into three main groups: seven cases with a PMI < 72 h; six cases with a PMI of 72−168 h and five cases with a PMI > 168 h. For each body, microbiological swabs were sampled from five external anatomical sites (eyes, ears, nose, mouth, and rectum) and four internal organs (brain, spleen, liver, and heart). Results: The HPM became increasingly different from the starting communities over time in the internal organs as well as at skin sites; the HPM microbiome was mostly dominated by Firmicutes and Proteobacteria phyla; and a PM microbial turnover existed during decomposition, evolving with the PMI. Conclusions: MALDI-TOF is a promising method for PMI estimation, given its sample handling, good reproducibility, and high speed and throughput. Although several intrinsic and extrinsic factors can affect the structure of the HPM, MALDI-TOF can detect the overall microbial community turnover of most prevalent phyla during decomposition. Limitations are mainly related to its sensitivity due to the culture-dependent method and bias in the identification of new isolates.

Variable levels of spike and ORF1ab RNA in post-mortem lung samples of SARS-CoV-2-positive subjects: comparison between ISH and RT-PCR.

Post-mortem examination plays a pivotal role in understanding the pathobiology of the SARS-CoV-2; thus, the optimization of virus detection on the post-mortem formalin-fixed paraffin-embedded (FFPE) tissue is needed. Different techniques are available for the identification of the SARS-CoV-2, including reverse transcription polymerase chain reaction (RT-PCR), immunohistochemistry (IHC), in situ hybridization (ISH), and electron microscopy. The main goal of this study is to compare ISH versus RT-PCR to detect SARS-CoV-2 on post-mortem lung samples of positive deceased subjects. A total of 27 samples were analyzed by RT-PCR targeting different viral RNA sequences of SARS-CoV-2, including envelope (E), nucleocapsid (N), spike (S), and open reading frame (ORF1ab) genes and ISH targeting S and Orf1ab. All 27 cases showed the N gene amplification, 22 out of 27 the E gene amplification, 26 out of 27 the S gene amplification, and only 6 the ORF1ab gene amplification. The S ISH was positive only in 12 out of 26 cases positive by RT-PCR. The S ISH positive cases with strong and diffuse staining showed a correlation with low values of the number of the amplification cycles by S RT-PCR suggesting that ISH is a sensitive assay mainly in cases carrying high levels of S RNA. In conclusion, our findings demonstrated that ISH assay has lower sensitivity to detect SARS-CoV-2 in FFPE compared to RT-PCR; however, it is able to localize the virus in the cellular context since it preserves the morphology.

A Clue on the Skin: A Systematic Review on Immunohistochemical Analyses of the Ligature Mark.

BACKGROUND: A ligature mark is a common injury in cases of hanging or strangulation. Estimation of age and vitality of the ligature mark can be crucial for differentiating antemortem and postmortem wounds and to distinguish between simulated suicidal hanging or accidental strangulation to conceal a crime and not simulated events. The immunohistochemistry has been recommended by several Authors as a reliable tool to determine whether an injury was sustained during life or not. Unfortunately, no general agreement on the immunohistochemical markers to be used has been found among the scientific community. The aim of the study was to detect the type and function of the immunohistochemical markers useful in the assessment of the vitality and age of the ligature marks for routine diagnostics. METHODS: Papers available on Pubmed, Scopus, and Web of Science were reviewed according to the PRISMA statement. RESULTS: Only eight papers satisfied all the following inclusion criteria: full texts in English dealing with human ligature marks and immunohistochemistry published on impacted or indexed scientific journals. CONCLUSIONS: The assessment of the vitality of a ligature mark is still a challenging topic in forensic science. Under ideal conditions and in compliance with autopsy protocols, the diagnosis of death by hanging or strangulation on fresh bodies can be better supported by autopsy findings other than a ligature mark. The validation of immunohistochemical markers on large series could be of help in doubtful cases and differential diagnoses.

Long-Term Detection in Hair of Zolpidem, Oxazepam and Flunitrazepam in a Case of Drug-Facilitated Sexual Assault.

Drug-facilitated sexual assault (DFSA) cases are pretty common in forensic toxicology. In this case report, a 56-year-old female tourist claimed to have been sexually assaulted by five men after having had a drug-spiked alcoholic drink. Urine samples were collected at 38, 44 and 45 h after the alleged rape. After 7 months, hair strands (28 cm in length) were also sampled to perform the segmental hair testing. The urine samples and decontaminated hair segments were tested for different groups of basic, acidic and neutral substances (γ-hydroxybutyrate or GHB, Z-drugs, barbiturates, benzodiazepines, hypnotics, antipsychotics and drugs of abuse). Gas chromatography-mass spectrometry and liquid chromatography-tandem mass spectrometry methods were applied for the qualitative and quantitative analyses. Toxicological analyses performed on the urine samples showed inconclusive findings. Zolpidem, flunitrazepam and oxazepam were detected in the hair segments corresponding to the time frame of the alleged assault. The endogenous levels of GHB were detected along the hair shaft. No drugs were detected in the proximal and distal hair segments or in washing solutions. This DFSA case demonstrated that the segmental toxicological analysis of hair, even when performed 7 months after the sexual assault, can provide evidence consistent with a single exposure to psychoactive drugs, at the time of the offense.