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.

Assessing wound vitality in decomposed bodies: a review of the literature.

The capability of discriminating between a vital and a post-mortem injury has always been a central theme in forensic pathology, particularly when the corpse is an advanced state of decomposition. Post-mortem decay of the body can mask or disrupt the classical features of a skin lesion, making it difficult to establish the cause and manner of death. Taphonomically challenging situations pose several interpretative issues of skin lesions which need to be addressed with scientifically recent methods that are still limited in the forensic literature. For that reason, the present research aims at resuming what is currently available in the attempt to provide some insight regarding this topic. This review considers only original researches, in which the markers of vitality were studied a significant amount of time after death, in order to test post-mortem persistency of these markers over time. A number of 132 original articles and reviews were considered, and the most significant results are resumed in an overview table and in two intuitive figures. Though many researchers tried to establish the vitality of lesions in specimen, few analysed samples from bodies when a significant degree of putrefaction or burning had occurred. The most significant marker proved to be GPA, which sowed a satisfying persistence over time (up to 6 months in air putrefaction and 15 days in water). However, what clearly emerged is that further studies are needed to address the challenges of taphonomically transformed specimen and to possibly neutralize the variability of experimental conditions, which affect the reproducibility of results. In conclusion, this study could be a starting point for providing food for thoughts about the most useful markers to search for in unusually tricky autopsy cases.

Intra-familiar homicides: From 2006 to 2021 in the judicial district of Milan.

INTRODUCTION: The present research aims at analyzing criminological and medico-legal characteristics of intra-familiar homicides occurred in two major Italian cities (Milano and Monza) from the beginning of 2006 to the end of 2021. METHODS: Cases were identified using the Institutional database of the Institute of Legal Medicine of Milan, where all the autopsies of victims were performed. Data about these cases were obtained from autopsies findings, preliminary investigation reports, local papers and victim relatives' interviews. In this period 11,480 autopsies were performed: 392 were homicides and, among these, 94 were confirmed as intra-familiar homicides (as a result of 84 events). Cases were classified according to the classic definition of intimate partner violence (referring only to an intra-familiar context), parricides, filicides, fratricides, familicides and grannicides. Age, sex, nationality and risk factors of the victims and perpetrators, as well as methods of murder were registered for each case. Only in some cases, motives for murder were known. RESULTS: The most frequent type of intra-familiar homicides was intimate partner violence (41.5%), followed by parricides (16%, mainly matricides), filicides (10.7%) and fratricides (6.4%). 9.6% of the total number of events were familicides. Risk factors were frequently involved, in particular among perpetrators and in the group of parricides, while among siblicides they showed minimal relevance. Overall, psychiatric pathologies were the main risk factors involved (at least 23.8%), unlike extrafamiliar homicides, where previous criminal report or involvement in illicit traffics are frequently reported among perpetrators. This finding enlightens the difficulties of caring for a relative with a mental disorder, who can become dangerous for the domestic caregiver. Furthermore, the high number of physical illness and the advanced age of victims points out the tragical consequences of the lack of social support system for these categories of people. METHODS: of murder were mainly cold steel (30-31.9%) and firearms (21-22.3%). However choking/manual ligature (9-9.6%), blunt force trauma inflicted using objects found on the scenario or bare handed (16-17%) and combined methods (11-11.7%) were frequently represented confirming the expected high level of impulsivity related to this kind of crime.

A retrospective study on femicides assessed at the Institute of Legal Medicine of Milan. Are older women at risk?

The purpose of the present research is to analyze temporal tend, victim's and perpetrator's specific features and the role of risk factors of femicides assessed at the Institute of Legal Medicine of Milan (in the North of Italy) from 2006 to 2020. The classical definition of femicide (the killing of a woman because of her gender) was used, in order to exclude murders of a woman because of economic issues, neighborhood quarrels, vengeance for previous discussions and other motives. Data were obtained from autopsies findings, local papers and victim relatives' interviews. The analysis of 86 femicides occurred in the judicial district of the Cities of Milan and Monza from 2006 to 2020 were performed. The results show the temporal distribution of the phenomenon, the methods of murders, the mean age of the victims, victims and perpetrators' relationship and nationality, risk factors. Although a reduction in the total number of femicides was observed, the percentage of femicides when compared to the total number of homicides was the same. Also, an increasing mean age of the victims over the period emerged. Moreover, the analysis of nationality of the victims and the perpetrators did not reflect foreign citizens distribution in Italy.

Sexual assault and abuse committed against family members: An analysis of 1342 legal outcomes and their motivations.

BACKGROUND: Over the past years medical centres specifically addressed in gender-based violence have developed protocols for the collections of evidence useful in the courtroom, including accurate documentation of physical and psychological states of the victim and collection of samples. Previous studies showed an association between documented physical trauma and conviction but unfortunately, few studies in the recent literature analysed the factors that influence the legal outcome and final judgement. The present study focused on the elements that appeared of significance in the legal outcome, including medico-legal evaluation, source of the crime report and circumstance of the assault. METHODS: It was conducted a retrospective analysis of all the judgments issued by the Public Prosecutor's Office at a Court of a Metropolitan Italian city regarding sexual and domestic violence, from January 1st 2011 to 31st December 31st 2015. Examination regarded the demographic information of the victim and of the defendant, information on the crime, the circumstances of the aggression and medical information retrieved. Sentences were subsequently divided into two categories based on the legal outcome (conviction vs acquittal) and the different characteristics of the two sub-populations were compared to verify if there were variables significantly associated to the judge's final judgment. RESULTS: Over the 5 years taken into consideration, there have been 1342 verdicts regarding crimes of sexual violence (374 cases) and regarding abuses against family members or cohabitants (875): other 93 cases regarded both sexual violence and abuse. 66.3% ended in conviction of the offender and 33.7% in acquittal of the accused. Cases of conviction were more frequent when they involved: use of a weapon by the assailant, as well as if the assailant had a criminal record and had a history of drug abuse or other addictions; duration of proceeding less 22 months and a civil party involved; presence of clinical documentation together with other deposition in addition to victim's deposition; also frequent episodes of violence and application of precautionary measures were associated to conviction. CONCLUSIONS: Many factors seem able to influence the judge's judgment, although clearly each case must be singularly evaluated. The mere presence of medical documentation, without the support of other sources of evidence, such as the victim's statement or further declarations, however, is almost always not definitive for the verdict. Despite so, in cases where there are multiple sources of evidence, clinical documentation can provide useful elements and can give clues on the consistency between the history told and injuries observed.

Male victims of sexual abuse and domestic violence: A steadily increasing phenomenon.

Violence perpetrated on male victims is a phenomenon that is currently underestimated by both national and international scientific communities, since males are historically (and stereotypically) considered the perpetrators rather than the victims of violence. As a consequence, the available literature lacks data which would allow a better understanding of this issue and its presenting features. We undertook a retrospective analysis of 231 medical files of male victims of violence over a five-year-period (2014-2018) at the Soccorso Violenza Sessuale e Domestica Centre in Milan, Italy. The sample included 112 victims of sexual violence and 119 victims of domestic violence, most of whom were younger than 18 years of age. The main aim of this study was to describe the presentations of male victims of violence in order to understand the phenomenon better, increase awareness of the issue and enable improved health-care management. The need to pay more attention to this vulnerable part of the population is mandatory in health-care services, and it includes: the provision of management guidelines, training to attending physicians and a supportive service to male survivors. Understanding which risk factors are related to male abuse can help with the development of programmes that identify, prevent and minimise violence - this being especially useful for primary-care clinicians. This is the first Italian study to deal with domestic and sexual violence involving male victims.

Drugs in bone: Detectability of substances of toxicological interest in different states of preservation.

In forensic contexts of advanced decomposition, when conventional matrices are no longer available for toxicological analyses, finding alternative matrices is necessary. The skeleton, which is fundamental for anthropologists and geneticists, could be useful also for toxicological purposes. The present study aims to examine what kind of information toxicological analysis performed on bones (the cranium and the ribs) in different states of preservation could provide to the forensic practitioner. Thirty cadavers with known pharmacological history, subjected to forensic autopsy at the Institute of Legal Medicine of Milan, were selected. Rib and cranium samples were collected from each body and separated into two parts in order to create two different states of preservation: One was cleaned from soft tissues and analyzed as a well-preserved bone sample; the other was submitted to a long maceration process, simulating complete skeletonization. All specimens were then processed with accelerated solvent extraction and the eluates analyzed using Q-Exactive™ Orbitrap™ Mass Spectrometer. The analysis of blood and skeletal matrices showed positive results for the tested substances in 63% of cases, mainly benzodiazepines, antidepressants, and drug abuse. Significant Pearson correlations were observed between non-macerated vs. macerated bone samples: r = 0.79 for rib samples, r = 0.61 for cranium samples, and r = 0.69 for all bone samples. As a consequence, the positive results confirm the potential of the bone tissue as an alternative matrix in forensic toxicology, even in cases of extremely decomposed bodies. This study also highlighted important elements for reconstructing the biological profile in cases of forensic anthropological concern.

Understanding the structural features of symptomatic calcific aortic valve stenosis: A broad-spectrum clinico-pathologic study in 236 consecutive surgical cases.

BACKGROUND: With age, aortic valve cusps undergo varying degrees of sclerosis which, sometimes, can progress to calcific aortic valve stenosis (AVS). To perform a retrospective clinico-pathologic investigation in patients with calcific AVS. METHODS: We characterized and graded the structural remodeling in 236 aortic valves (200 tricuspid and 36 bicuspid) from patients with calcific AVS (148 males; average 72years); possible relationships between general/clinical/echocardiographic characteristics and the histopathologic changes were explored. Twenty autopsy aortic valves served as controls. In 40 cases, we also tested the immunohistochemical expression of metalloproteinases and cytokines, and characterized the inflammatory infiltrate. In 5 cases, we cultured cusp stem cells and explored their potential to differentiate into osteoblasts/adipocytes. RESULTS: AVS cusps showed structural remodeling as severe fibrosis (100%), calcific nodules (100%), neoangiogenesis (81%), inflammation (71%), bone metaplasia with or without hematopoiesis (6% and 53%, respectively), adipose metaplasia (16%), and cartilaginous metaplasia (7%). At multivariate analysis, AVS degree and interventricular septum thickness were the only predictors of remodeling (barring inflammation). All the tested metalloproteinases (except MMP-13) and cytokines were expressed in AVS cusps. Inflammation mainly consisted of B and T lymphocytes (CD4+/CD8+ cell ratio 3:1) and plasma cells. AVS changes were mostly different from typical atherosclerosis. Cultured mesenchymal cusp stem cells could differentiate into osteoblasts/adipocytes. CONCLUSIONS: Structural remodeling in AVS is peculiar and considerable, and is related to the severity of the disease. However, the different newly formed tissues-where "valvular interstitial cells" play a key role-and their well-known slow turnover suggest a reverse structural remodeling improbable.