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In pathological-forensic literature blast injuries are usually studied in relation to suicides, homicides during terroristic attacks and accidental events on the workplace, while the finding of these kinds of fatalities during misdoing is a recent occurrence. The Authors describe the peculiar case of the accidental death of the member of a criminal mob specialized in thefts to ATMs using acetylene tanks. The victim, having poured out the acetylene with the intent of triggering an explosion, was involved in the blast of the tank, deceasing immediately. This unusual manner of death highlights the danger of room saturation with acetylene, a criminal technique becoming more and more popular in the last years. [ABSTRACT FROM AUTHOR]

In a forensic scenario, if biological stains are found in very small quantities, these are usually collected for DNA analyses, considered essential for the forensic investigation and thus excluding possible investigations by other forensic disciplines as forensic toxicology. We developed an experimental study to evaluate the feasibility of analyzing DNA extraction residues obtained from DNA extraction procedures to perform toxicological analysis, with the aim to extract both genetic and toxicological information without affecting or compromising the genetic sample and/or DNA extraction. DNA extraction from four blood samples (fortified with 5 molecules of interest with a final concentrations of 1 µg/mL, 100 ng/mL, 10 ng/mL and 5 ng/mL, respectively) were analyzed with QIAGEN QIAmp® DNA Mini kit. Three waste residues collected from the DNA extraction were analyzed for the toxicological investigation via Solid-Phase Extraction and High-Performance Liquid Chromatography-Tandem Mass Spectrometry analyses (Thermo Scientific™ TSQ Fortis™ II Triple-Quadrupole Mass Spectrometer). The analytical investigation revealed that our analytes of interest were detected in two different residues of the DNA extraction procedure, allowing both genetic and toxicological analyses without affecting the DNA identification. At last, the experimental protocol was applied to a hypothetical case, with encouraging results and allowing the identification of our molecules of interest., (© 2023. The Author(s).)

Introduction: In this study we microscopically investigated, for the first time ever, the colorimetric detectability of barium of gunshot residues (GSR) on cadaveric human skin with gunshot wounds., Methods: For this purpose we used two different colorimetric techniques known in the literature, namely 0.2% sodium rhodizonate (Na-R-Ba 0.2%) and sodium rhodizonate in alcoholic environment (Na-R-Ba OH 0.2%). At the same time, we have also coupled it with scanning electron microscopy with energy dispersive X-ray (SEM/EDX) analysis and the colorimetric study for the detection of lead of GSR. These techniques were applied to 16 victims who died from gunshot injuries, as well as to a control group., Results: SEM/EDX demonstrated the presence of lead in all cases and barium in 11 of the 16 cases. The subsequent colorimetric technique with Na-R-Ba 0.2% did not show the barium of GSR in any case, unlike the Na-R-Ba OH 0.2% technique. This latter, in fact, has demonstrated the presence of this metal in 2 cases (18%). No microscopic case of false positive was recorded., Conclusion: The evidence obtained with Na-R-Ba OH 0.2% makes this method, applied here for the first time ever, worthy of further study. Meanwhile, although this technique can certainly be applied, it cannot be separated from the contextual colorimetric investigation for lead and the use of more sophisticated techniques., Competing Interests: The authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article., (© The Author(s) 2024.)

Abstract: Paraquat (PQ) is one of the most widely used herbicides in the world, and poisoning is generally associated with accidental, suicidal, or homicidal events. Therefore, in the forensic context, PQ could be in various ways involved as a possible cause of death of a subject. However, even though its systemic toxicity is known, the biological effects exerted on individual viscera have been explored only to some extent, especially in case of victim's survival. Therefore, a case concerning a suicidal ingestion of PQ with survival of 3 days was deemed of interest. Clinical toxicological analyses confirmed acute PQ intoxication, and after the death of the victim, an autopsy was performed showing local and systemic signs of ingestion of a caustic substance. Histologic examination revealed marked cellular damage to the major viscera, particularly the gastroesophageal tract, liver, kidneys, and lungs, with initial alveolar fibrosis noted despite the patient's short survival. This finding represents a new element in the context of PQ lung injury, as it has not been previously documented in the literature. Thus, histological findings in lethal intoxications after survival can reveal specific peculiarities still unknown and, therefore, assume transversal relevance not only at forensic but also clinical level., Competing Interests: The authors report no conflict of interest., (Copyright © 2023 The Author(s). Published by Wolters Kluwer Health, Inc.)

A suicide where the deceased has employed more than one means of ending his or her life is defined as a complex suicide (CS). Forensic practitioners may face difficulties caused by the articulated mechanisms underlying this event. Among CS, the combination of hanging and gunshots is unusual. In this study, we present three unique cases of such planned complex suicides (PCS) that we have encountered in our 28 years of activity at the Bureau of Legal Medicine of Milan. Careful inspection of the death scene, precise analysis of the anamnestic-circumstantial data, and accurate medico-legal autopsy examination were the starting points for a better understanding of the causes and manner of death. In particular, the presence of vital reactions of tissues involved in the two different means used, the coherence of the areas involved with a self-inflicted wound, and the absence of signs of third party intervention allowed us to classify these events as suicides. As for the chronology of events, the lethality of the cerebral lesions caused by the gunshots in all cases, in accordance with the cervical lesions caused by hanging, led us to conclude that we were dealing with PCS and catalogue these three cases as unusually planned complex suicides avoiding incorrect and superficial classification., Competing Interests: Declaration of conflicting interestsThe authors declare no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Competing Interests: Conflict of interest: None.

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., Competing Interests: Declaration of competing interest The authors declare that they have no conflict of interest., (Copyright © 2023 The Authors. Published by Elsevier Ltd.. All rights reserved.)

In contrast to other forensic disciplines, forensic microbiology is still too often considered a "side activity" and is not able to make a real and concrete contribution to forensic investigations. Indeed, the various application aspects of this discipline still remain a niche activity and, as a result, microbiological investigations are often omitted or only approximated, in part due to poor report in the literature. However, in certain situations, forensic microbiology can prove to be extremely effective, if not crucial, when all other disciplines fail. Precisely because microorganisms can represent forensic evidence, in this narrative review all the major pathological forensic applications described in the literature have been presented. The goal of our review is to highlight the versatility and transversality of microbiology in forensic science and to provide a comprehensive source of literature to refer to when needed., Competing Interests: Declaration of competing interest The authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article., (Copyright © 2023 The Authors. Published by Elsevier Ltd.. All rights reserved.)

In forensic toxicology, when conventional matrices are no longer available, alternative matrices can be used to assess toxicological investigations. Clothes worn by skeletal remains may be a good unconventional matrix for toxicological analyses considering that they have absorbed decomposition fluids and blood from a body. We hypothesized a scenario in which a skeleton, wearing clothes, was discovered in an open environment. From this starting point, an experimental study was developed on different textiles (cotton, wool, and polyester) to evaluate whether water-related atmospheric agents and molecule solubility can largely influence the detection of molecules of toxicological interest on this specific matrix, together with the characteristics of different garments chosen. The experimental study was performed on blood spots, previously spiked with 6-monoacetylmorphine and morphine, accurately placed on different textiles and washed with different quantities of deionized water adjusted at pH 5.6 with formic acid to simulate different rainfall conditions. Toxicological analyses were performed via Solid-Phase Extraction and High-Performance Liquid Chromatography-Tandem Mass Spectrometry analyses (Thermo Scientific™ TSQ Fortis™ II Triple-Quadrupole Mass Spectrometer). From the experimental study morphine could not be detected on 100% cotton and 100% wool fabric after the passing of 500 mL of deionized water and in 100% synthetic polyester textile after washing with 250 mL of deionized water. In conclusion, when toxicological analyses are carried out on unconventional matrices as textiles worn by corpses exposed to different environmental conditions, it is of great importance, in using such substrates as evidence for the presence of molecules of toxicological interest, to evaluate chemical-physical characteristics of each analyte under investigation in order to correctly interpret the toxicological data obtained., (© 2023. The Author(s).)

Background: Suicide methods chosen by victims are particularly critical in suicide risk research. To differentiate suicide deaths, it is usual to categorize them as violent and nonviolent depending on the detrimental method chosen by the victims. Caustic ingestion, for example, is traditionally considered as a nonviolent suicide method. It results in severe consequences for the human body and it is associated with high levels of lethality., Methods: In this study, we retrospectively analyzed suicides that occurred between 1993 and 2021 in Milan (Italy) and that underwent autopsy. We compared a sample of 40 victims that ingested caustic substances with a sample of 460 victims of other chemical ingestion, and a sample of 3962 victims from violent suicide. Univariate analyses and univariate logistic regression models were performed. Suicides from caustic poisoning were significantly older, had a higher mean number of diseases and were more affected by psychiatric diseases compared to other chemical ingestion victims. By contrast, caustic suicides, compared to violent suicides, had a more balanced gender ratio, a higher mean number of diseases, were more affected by psychiatric diseases, had a higher rate of complex suicides (more than one modality), and had victims who died more frequently inside instead of outside. In logistic regression models, age was the only feature differentiating caustic from other chemical ingestion suicides while the features differentiating caustic from violent suicides were gender, mean number of diseases and suicide place., Conclusions: Suicides by caustic ingestion showed substantial differences compared to violent suicides, with a higher severe profile. However, some differences were reported comparing caustic ingestion to other chemical ingestion as well. Thus, we argue whether it is more appropriate to differentiate the suicidal ingestion of caustics from both violent and nonviolent suicide methods.

The term "cocoon syndrome" defines a sclerosing encapsulating peritonitis (SEP) that involves a chronic fibrotic inflammatory reaction of the parietal peritoneum and of the viscera leading to a complete sclerosis. The cocoon that is formed causes an incarceration of the intestinal loops with severe complications leading to high mortality. We are presenting the case of a 15-year-old young man that underwent surgery for appendectomy and that was evaluated for having a regular abdominal state. During the post-surgery period, however, several episodes of intestinal occlusion required further surgical interventions leading to a right hemicolectomy. The presence of a fibrotic-adhesive ligneous peritonitis with blended intestinal loops, severely thickened walls, and intestinal scaring stenosis was observed during his second surgical operation. A stenosis of the colostomy led to a worsening of the vital signs of the young man with the onset of a cardiac failure and subsequent decease. Macroscopic autopsy examination and histological analysis confirmed the severe obstructive adhesive encapsulating abdominal context allowing to trace back the cause of death to a cocoon syndrome. Since no predisposing factor could be found, we hypothesized that this case could be characterized by an excessive peritoneal reactivity due to surgical appendectomy. Cocoon syndrome is a rare pathology, and its microscopic features are seldomly observed and could be underestimated. We present a directly observed case with a very substantial macroscopic and microscopic context., (© 2022. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Competing Interests: The authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

In this paper, we present the results of toxicological analyses of preserved brain tissue and bone samples from the remains of the seventeenth century patients of the Ospedale Maggiore, the main hospital in Milan and one of the most innovative hospitals in Europe from the Renaissance period. Beneath it, the crypt functioned as the burial place for the deceased of the hospital. In this multidisciplinary study of the remains, toxicological analyses in particular were performed with HPLC-MS/MS on different biological samples from nine individuals. Anthropological, paleopathological, histological, radiological examinations and radiocarbon dating were also carried out. As a result, archeotoxicological analyses revealed the presence of codeine, morphine, noscapine and papaverine, derived from Papaver somniferum, a plant present in the hospital pharmacopeia used as a narcotic, analgesic, astringent, coagulant, and antitussive agent. Such analyses have shed light on the pharmacological therapies administered to the patients near the time of death and have implemented our knowledge of medical treatment and drug administration in the 1600's., (© 2023. The Author(s).)

Nowadays, the diagnostic value of postmortem microbiological investigations is still a debated topic, but postmortem microbiology (PMM) remains a discipline with great forensic potential. To evaluate the usefulness and diagnostic-forensic value of postmortem microbiological cultures, it has been conducted a study on cadaveric material sampled during autopsy aiming to identify the correct cause of death. The study analyzed 45 cadavers subjected to judicial autopsy, divided into two groups based on the presence or absence of external or internal macroscopic autopsy signs suggesting infectious pathology. In the same cases, both the microbiological and conventional histological investigations have been simultaneously carried out. From the investigations, mono-bacterial, mono-fungal, mixed and negative cultures were observed. In mono-species microbiological growth, the histological epicrisis confirmed an infectious cause of death due to the presence of signs of acute infection with an aggressive infectious agent. In cases where growth was mixed, it was possible to distinguish between simple postmortal contamination and perimortem colonization. Finally, in some cases where the microbiology was negative, this has been essential in highlighting signs of a vital reaction to viral or parasitic infection. The joint and integrated evaluation of the laboratory results made it possible to correctly understand even those peculiar situations in which the PMM results alone would not have been significant. These methods, when combined, constitute an optimal forensic approach for the identification of the real cause of death and thus reduce the number of unsolved cases., Competing Interests: Conflicts of interest/Competing interests The authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Synthetic cathinones are a class of psychoactive drugs that have become, in recent years, of quite common observation in medical and toxicological forensic field. MDPHP (3,4-methylenedioxy-α-pyrrolidinohexanophenone) belongs to this class of substances but lethal acute intoxication caused by this specific substance without other co-ingestions has never been described in literature. We are presenting the unique case of a fatal acute intoxication caused by MDPHP in a 48-year-old man carried to the A&E department of a big Milanese hospital where he suddenly deceased after brief time. Clinical records, autopsy examination, histological findings, and toxicological analysis, assessed via a Q-Exactive Orbitrap with a HPLC system and LC/MS-MS system, are discussed. This case report may represent the first case of this type of intoxication and might help in improving recognition and treatment of these particular cases., (© 2022. The Author(s).)

Microscopic examination of mummified or corified skin may be of extreme importance for forensic purposes. However, standard histological samples in these cases are low-end, and preparation is burdened by several problems and so are diagnostic results: an improvement of these types of specimens is therefore advantageous. This study aims to identify the best performing rehydration solution among a fabric softener, a body lotion, and Sandison's rehydrating solution. Samples of skin undergoing mummification or corification were collected from 25 corpses and each sample was divided into 4 fragments: one of these fragments was directly fixated in 4% formalin, one was previously treated with a tissue softener, another one was previously treated with a body lotion, and the last one was treated with Sandison's solution. After 72 h, the pretreated samples were post-fixated in 4% formalin and then prepared for standard histological examination staining the histological slides with hematoxylin-eosin and Masson's trichrome. At the microscopic examination, samples directly fixated in formalin were characterized by usual marked structural alterations and altered stainability, typical of such dry tissues. Vice versa, those previously treated appeared to be better-preserved even though with different improvement levels: body lotion made a medium-low-grade restoration of the tissues, and fabric softener a high-grade restoration, while Sandison's rehydrating solution produced an optimal grade restoration. Sandison's rehydrating solution was confirmed to be the best rehydrating substance for mummified and corified skin. Fabric softener could be, however, considered a valid substitute, being productive of high-grade microscopic yield., (© 2022. The Author(s).)

Purpose: Since the solely certain remnants of a performed autopsy are formalin-fixed paraffin-embedded (FFPE) samples, stored in the archives of every institute of legal medicine, we managed to extract molecules of toxicological interest from these specimens., Methods: We assessed the analysis of ten fresh liver samples collected from heroin-related deaths and then histologically processed the same samples. The embedded blocks were then extracted by means of a new extracting method and the eluates were measured. We also selected five toxicological cases of heroin-related fatalities that were examined 20 years ago, collected the toxicological result documents of the analysis that were carried out at the time and then processed the corresponding FFPE liver samples that were stored in the archives., Results: We managed to isolate heroine-related metabolites from 20-year-old paraffin-embedded blocks and calculated ratios to evaluate the performance of our new extraction., Conclusions: According to our study, it is feasible to carry out a toxicological examination on old histological samples and, therefore, this matrix can be considered as a new alternative specimen for chemical-analytical evaluations of past cases or when fresh samples are not available anymore. The new extractive method was evaluated as efficient in treating these complex, paraffin-embedded samples. It was surprising that the target compounds could be quantitated from FFPE bocks created as long as 20 years ago., (© 2022. The Author(s).)

We are presenting a study on 136 cases performed in a 2-year period (2018-2019) at the Bureau of Legal Medicine of the University of Milan for which toxicological analyses were requested and we are making a detailed interpretation of clinical records and discussing toxicological results from each case included in the study. Total number of autopsies was 1323 and in 10.3% of the cases, toxicological analyses were requested to obtain further information. Analyses were assessed with High-Performance Liquid Chromatography-Mass Spectrometry system and Gas Chromatography-Mass Spectrometry analyses. Additionally, Blood Alcohol Concentration and detection of volatile substances were obtained with Head Space-Gas Chromatography-Mass Spectrometry system. From these analyses, 101 cases out of 136 provided positive results (74.3%). Main substances detected were cocaine, diazepam, morphine, and ethanol. The most representative profiles of individuals that emerged from this study were: a Caucasian male, age 41-50, that died for cocaine acute intoxication or was killed; a Caucasian male or female with a range-of-age of 31-50 deceased for simple suicide caused by acute intoxication or by complex suicide caused by acute intoxication and suffocation; and finally, a Caucasian male with a range-of-age 21-40 that died in a car accident without any toxicological evidence. From the results, acute intoxication at the time of death was confirmed in 54 cases and in 57 cases the toxicological analyses helped in the determination of the cause and manner of death. From this study, the importance of toxicological data among forensic sciences is confirmed., (© 2022 The Authors. Journal of Forensic Sciences published by Wiley Periodicals LLC on behalf of American Academy of Forensic Sciences.)

Cocaine is among the illicit substances most frequently implicated in deaths related to the use of drugs of abuse both worldwide and in Italy. Cutting agents involved in the adulterations of this substance are many, and the process of lacing can take place at various stages of the production of the drug. In this report we are discussing the case of a 27-year-old woman found dead next to her car in a wooded area in the suburban area of Milan. On the crime scene, several specimens of white powder were collected and subsequently analyzed via Q-Exactive Orbitrap with an high performance liquid chromatography system and liquid chromatography--tandem mass spectrometry analysis along with biological matrices sampled during autopsy examination. The toxicological analysis revealed that the death could be ascribed to a lethal dose of methomyl, a carbamide pesticide used as cutting agent for cocaine. According to literature, this is the first time that this substance is used as an adulterant., (© The Author(s) 2021. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Suicidal ingestion of food which the victim is aware they are allergic to is an exceptional occurrence in the forensic field. To the best of our knowledge, no cases of suicidal food anaphylaxis have been reported to date. Therefore we present the first case described in the literature. A 30-year-old prisoner was found dead inside his cell with the remains of a peach remains next to his body, and a handwritten farewell note in his pocket. The autopsy revealed only non-specific findings, while laboratory investigations (serological, toxicological, histological, and immunohistochemical) played a pivotal role in determing the cause and manner of death. In particular, a high titer of both total and specific IgE antibodies was detected, as well as an increase of the tryptase level in cadaveric blood. Moreover, a massive concentration of salicylates was measured in the gastric contents. Microscopically, cellular residues characterized by a vegetal structure were observed in the gastric contents and elements suggestive of mast cells were detected in the glottis, lungs, and myocardium. The immunohistochemical investigation with anti-CD117 and anti-tryptase antibodies showed positivity for mast cells, some of which appeared degranulated. Such findings were entirely consistent with an acute systemic anaphylactic reaction triggered by allergy. Therefore, the prisoner's death was attributed to self-induced food anaphylaxis caused by the ingestion of peaches. This conclusion was achieved based only on circumstantial data, anamnestic information, autopsy findings, and multiple laboratory results. This integrated approach should be used to pursue a post-mortem diagnosis of anaphylaxis., (© 2021. The Author(s).)

The microscopic examination of a hanging cutaneous furrow, stained with Goldner's Masson trichrome staining, highlighted an abnormal dyeing inversion affinity of the connective tissue - red instead of green - located in the region of maximal cutaneous compression and exsiccation. To identify if this different stainability could be considered as an intrinsic characteristic of all biological tissues compressed and exsiccated, we have assessed different cadaveric skin samples that underwent traumatic detrimental phenomena that can produce such effects. We collected skin fragments from 24 corpses deceased because of gunshot injuries, electrocution, hanging, and heat-induced lesions, sampling the areas directly involved, as well as skin specimens to use as a control. The slides were stained with hematoxylin and eosin and two different protocols of Goldner's Masson trichrome staining (one homemade and one commercial kit). The inversion of the staining affinity of the connective tissue was observed in 83% of the cutaneous samples, using both the Goldner's Masson trichrome staining protocols. This phenomenon was not observed in any of the control cases. Therefore, the inversion of the staining affinity of the connective tissue dyed with Goldner's Masson trichrome staining may represent a histomorphological aspect that must be expected when the skin has been affected by specific detrimental modalities able of producing compression and exsiccation., (Copyright © 2021 Elsevier B.V. All rights reserved.)

We analysed the recorded cases of suicides committed other than by hanging in prison in Milan. A retrospective analysis was conducted on 25,512 autopsies performed from 1993 to 2019, selecting all the suicides in prison but our attention was focused solely on cases where an alternative mode to hanging was used. From a total of 97 suicide events in prison, 15 were consistent with the established search criteria: 6 victims died from plastic bag suffocation, 4 by direct inhalation of butane gas, 2 associated plastic bag suffocation to inhalation of butane gas, one committed suicide with an edged weapon, one by self-burning and, finally, one by voluntary ingestion of a food to which he was allergic, with the intent of inducing an anaphylactic shock. Our analysis has shown that the landscape of prison suicides is diverse, not limited solely to hanging. Therefore, it is necessary for the forensic scientific community to raise awareness of potentially unusual suicide methods in prisons and, in the same way, for the Penitentiary Administration to put adequate preventive measures and strategies in place.

We present a case of a 48-year-old man's unexpected death affected by a relapsed clivalchordoma. After partial excision surgery of the neoplasm, he manifested 5 days later, in conditions of well-being, a sudden lethal extracranial hemorrhage from nose and mouth. The autopsy examination and the subsequent histological investigations did not allow us to clarify the exact origin of the bleeding. Based on the negativity of the accurate examinations performed, the extent of the bleeding, and the findings highlighted by the means of the nuclear magnetic resonance (NMR) carried out a few days before death, we have considered reasonable to localize the source of hemorrhage in the intrapetrous tract of the left internal carotid artery. Since this is a unique event, never previously documented, we believe that our report may be of interest to the scientific community., Competing Interests: None

We undertook a retrospective analysis of deaths that took place in prisons in Milan between 1993 and 2017, by identifying cases from a total of 24,101 autopsies that were performed at the Section of Forensic Medicine of the University of Milan. From the archives of this institution, we found 227 autopsy reports relating to deaths that had taken place in one of Milan's three detention facilities. These deaths were divided into two types: natural deaths ( n =135; 59.5%) and violent deaths ( n =92; 40.5%). The groups have different characteristics: while natural deaths mostly resulted from cardiovascular diseases, suicides were mainly the result of hanging. Further, people who died by suicide often had a history of psychiatric disease and/or drug abuse, and over a quarter of them had previous suicide attempts and/or had declared suicidal intentions. This study confirms the need for good quality healthcare services for prisoners, given that they remain a population at high risk of early death.

Scanning electron microscopy with energy-dispersive x-ray (SEM/EDX) analysis is an investigation whose potential has become increasingly important in the field of forensic research and diagnosis. We present the procedure to perform a well-carried-out SEM/EDX analysis on corpses affected by different types of injuries, such as blunt force trauma, ligature strangulation, electrocution, sharp force trauma, gunshot wounds, and intoxication. After the areas of forensic interest have been macroscopically identified, the sampling can be performed in 2 different ways: apposition of the double-sided graphite tape on the damaged area or performing the excision of a biological sample. In both cases, a proper negative control sample is required. In all cases, SEM/EDX analysis can detect exogenous microtraces consistent with the types of injuries involved. In blunt force trauma, microparticles of different nature deriving from the contact of the blunt instrument with the victim may be observed; in sharp force trauma, metal microtraces (Fe, Cr, Al, Ti) can be identified. In ligature strangulation, exogenous microtraces may be found in the cutaneous furrow. In electrocution, it allows to identify the pathognomonic metal pattern (Cu, Zn, Fe) of the "electric mark." In gunshot wounds, the main applications regards the detection of metal particles (Pb, Ba, Sb) of gunshot residues. Finally, in the analysis of intoxicants, it may identify traces of toxic substances. Thus, the authors conclude that SEM/EDX analysis can provide essential information to assist in the medicolegal investigation of death.

In this study we have evaluated the detectability of lead deriving from Gunshot Residues (GSR) on human bone using colorimetric techniques as well as the possible interferences of decalcification on the stainability of these particles. 22 well preserved cadavers were divided into two groups: group A enlisted 11 victims deceased because of gunshot injuries (bones and soft tissues sampled in correspondence of the entry wound of the bullet)- and group B, comprehending 11 victims deceased for natural causes (specimens sampled from intact skin areas). All the samples, after decalcification processes for bone specimens with a 14 % HCl solution, underwent standard histological procedures and were stained with hematoxylin and eosin (H&E), sodium rhodizonate (Na-R) and acidified sodium rhodizonate (Na-R-HCl 5 %). Two additional slides of soft tissues for each group were pre-treated with HCl 14 % before being stained with likewise staining. In group A, 10 samples out of 11 resulted to be negative to Na-R and Na-R-HCl 5% techniques while all the corresponding soft tissues resulted to be positive for the same staining. Additional soft tissue slides processed with 14 % HCl solution were negative in 10 cases out of 11. No sample in group B showed positivity to the staining procedures neither with nor without pre-treatment with 14 % HCl. The search for GSR residues on human bone is therefore not executable with Na-R and Na-R-HCl 5 % techniques since the essential decalcifying procedure modifies the stainability of the residues therefore adulterating the results., Competing Interests: Declaration of Competing Interest The authors report no declarations of interest., (Copyright © 2020 Elsevier B.V. All rights reserved.)

The term overkill identifies the infliction of massive injuries that greatly exceed those needed to kill the victim both in number and intensity. We present the case of a Peruvian transsexual, who was choked and hit by 11 sharp-force wounds all in the facial area. The scanning electron microscopy with energy dispersion X-ray analysis carried out on the skin of the facial wounds proved crucial in proving that two different point-and-edge weapons were used, following the finding of metallic micro-traces of heterogeneous composition. This discovery and the autopsy highlighted the use of two distinct types of injury (mechanical asphyxia and sharp-force trauma) showed that this murder amounted to o verkill .

Microscopic study and interpretation of skin samples obtained from mummified/corified cadavers request the adoption of special laboratory techniques in order to avoid that the histological samples may detach from the slide during staining processes. We have assessed an experimental and extended drying of corified and mummified skin samples in order to evaluate an eventual improvement after staining procedures. Samples of skin undergoing different post-mortem putrefactive phenomena-from parchment-like to mummification-were collected from 16 corpses and divided in halves: one half was fixated in buffered formalin while the other half was previously treated with Sandison's rehydrating solution. The so-obtained sections were subsequently dried for one night at 37 °C and stained with Hematoxylin & Eosin staining and Masson's Trichrome staining according to Goldner. At microscope observation, 69% of the dyed sections stained with H&E and 53% of the sections stained with Masson's Trichrome was lost during different steps of the coloring procedures. We then proceeded with a new cutting of the slides and decided to experimentally dry them for 5 days at a mean temperature of 37 °C: purpose of this procedure was to gradually remove the possible liquid component of the samples probably culpable of the detachment. With this modified procedure, all the sections resulted to be preserved in the following staining steps and showed, moreover, an improvement of the morphological structure, especially in those previously treated with Sandison's rehydrating solution. In those cases where a histological examination is requested on skin samples collected from corified or mummified cadavers, this technique is highly recommended in order to obtain microscopic slide of high quality.

In electrocutions, death may be caused by alterations in the heart conduction system provoking ventricular fibrillation. This study aims to identify histological cardiac markers of high- and low-voltage electrocution. Two groups of decedents were evaluated: group A included 14 fatalities caused by high- or low-voltage electrocution and group B (control) included 14 fatalities due to other traumatic or disease causes. Myocardial sampling with microscopic examination was performed on all the hearts using the hematoxylin and eosin and Masson's trichrome stains to investigate morphological characteristics that could indicate the damage caused by high- and low-voltage electrocutions. Interstitial myocardial hemorrhagic infiltration was the only differentiating finding, which was shown only in high-voltage electrocution. This pathological finding has not been previously reported, and it may be specific to high-voltage electrocution deaths. Further studies are warranted., (© 2019 American Academy of Forensic Sciences.)

In a previous study, we presented a case of an elderly woman's sudden death, in which microscopic examinations showed intramyocardial eosinophilic material suspected for amyloid, but not definable as such to the classic Congo Red staining. To overcome the arisen interpretative and diagnostic difficulties, we experimentally modified the classic Congo Red staining, using a specific one for corpse. The finding of a low-intensity positivity allowed us to formulate a very likely diagnosis of occult lethal cardiac amyloidosis. However, this low-intensity positivity obtained after having applied this experimental method for the first time and in only one case, as well as the existence of the rare pathology known as microfibrillar cardiomyopathy, which may be related to the observed microscopic findings, have forced us to investigate the correctness of the diagnosis. For this purpose, we performed in-depth investigations with sodium sulphate-Alcian Blue (SAB) staining and immunohistochemistry. Thanks to them, the amyloid nature of the intramyocardial material was confirmed and has been proved not only the reliability of our experimentally modified technique, but also the appropriateness of the diagnosis previously formulated. Therefore, the supposed involvement of the microfibrillar cardiomyopathy was excluded., Competing Interests: Declaration of competing interest The authors report no conflicts of interest. The authors alone are responsible for the content and writing of this paper., (Copyright © 2020 Elsevier Ltd and Faculty of Forensic and Legal Medicine. All rights reserved.)

We present a retrospective study of prostitutes' homicide in the Milanese area over a 26-year period (1993-2018), and compare the results with general women's homicides: 294 women were killed of whom 71 were prostitutes. In the general population, the type-victim is an Italian woman aged between 31 and 40 years suffering from no particular pathologies or drug dependence. Prostitutes are 10 years younger, mostly Italian, suffering from pathologies probably related to their activities. Prostitutes remain at high risk of violence, due to gaps in the crime prevention system.

The banning of the heavy metal thallium (Tl) in many Countries, because of its toxicity, led to a remarkable reduction of the number of cases of poisoning both accidental and homicidal forcing us to better study the pharmacokinetics of this poison using new technologies. The Authors, in this work, are reporting the case of a collective thallium toxicosis caused by voluntary adulteration of an infusion with thallium sulfate, occurred in 8 members of the same familial nucleus; the administration of Prussian Blu resulted to be ineffective for 3 of these members that died at a later time. The most peculiar aspects of this rare manner of poisoning are discussed; the analytical procedures used, Inductively Coupled Plasma Mass Spectrometry (ICP-MS) in particular, resulted to be fundamental in the forensic diagnosis process of acute poisoning cause by thallium., (Copyright © 2019 Elsevier B.V. All rights reserved.)

In forensic evaluation of charred corpses, internal detrimental signs may result as more significant of those observed during external examination and is often arduous to state if a victim was exposed to fire before or after death. When the histological analysis of the remaining internal viscera is necessary, the massive destruction caused by the lesion, the charring and the coarctation of the samples don't allow to give further information or to determine the remaining organic components of the viscera. This limit is determined by the intrinsic characteristics of this thermal lesivity of self-maintenance even after the exitus of the subject, worsening the initial detrimental framework. The Authors, with the purpose of improving the microscopic visualization of the samples collected from cadavers with peculiar deterioration, as in case of carbonization, suggest the use of a specific technical protocol based on the use of Sandison's rehydrating solution since the samples treated with this solution showed, at microscopic examination, a substantial histological-morphological improvement., (Copyright © 2019 Elsevier B.V. All rights reserved.)

The retention of gauze within the mediastinum is considered an exceptional finding. We are presenting the case of a woman, affected by aorto-mitral valvulopathy, in whose mediastinum was found, during the autopsy, a gauzome without signs of active phlogosis, which had been "forgotten" in a previous surgery, acted 14 years before the death. A 39 y.o. woman had received surgical treatment of double aorto-mitral valve substitution and tricuspid plastic. At age 53, she successfully received surgery once again at a different hospital; she then started rehabilitation therapy, during which she deceased due to sudden hematemesis with infectious multi organ failure. During the autopsy, a 5 cm long surgical gauze was found in the mediastinum. A SEM/EDX (Scanning Electron Microscopy with Energy Dispersive X-ray spectroscopy) exam was ran on the foreign body and a sample of the same gauze used in the cardiac-surgical department of the hospital the victim had passed away in, to act a comparison between the two kinds of material. The exam showed a morphological difference between exogenous fibers incorporated within the gauzome and the control gauze. This case highlights the diagnostic and interpretative challenges in finding a foreign body within the mediastinum without signs of active phlogosis. SEM/EDX investigations executed on the gauzome and a "control gauze" allowed the attribution of malpractice to the first hospital the victim had been operated in 14 years before the exitus. The SEM/EDX exam confirms its importance in correctly identifying the case, which becomes an example for similar happenings., (Copyright © 2019. Published by Elsevier B.V.)

Literature on electrocution in Italy is limited. The authors have focused their attention on the use of scanning electron microscope/energy-dispersive X-ray analysis in order to evaluate the current mark even on paraffin-embedded samples. A total of 24,104 autopsies were performed at the Section of Legal Medicine of Milan (1993-2017); all cases of death caused by electrocution (low and high voltage) were selected. We assessed a regular histological examination, a toxicological examination and detected metallisation with energy-dispersive X-ray spectroscopy analysis on paraffin-embedded tissues of typical current marks. We collected 27 high-voltage fatalities and five cases of low-voltage suicide electrocution; the technique revealed itself as sensitive. In all cases, microscopic examination highlighted the typical signs of electric current and thermal damage but the histochemical specific stainings for copper and iron gave negative results. Scanning electron microscope/energy-dispersive X-ray analysis, assessed on these same samples, even when paraffin-embedded, however, enabled us to detect these elements and differentiate between the various metal residues involved in the electrocution process. Based on their experience, authors advise the use of scanning electron microscope/energy-dispersive X-ray analysis on electrocution deaths to provide information for forensic diagnosis.

The authors used a particular protocol on putrefied corpses to highlight the cutaneous furrow. Two groups of cadavers were selected: in the first group (suicide by hanging mechanical asphyxia), the authors sampled cutaneous lozenges on the furrow still macroscopically recognisable, while in the second group (corpses deceased by different means), we sampled cutaneous lozenges in the hypostatic leaning. All specimens were divided in two: one underwent standard fixation while the other, previously rehydrated in Sandison's solution, was then fixed in formalin. All specimens were stained with hematoxylin and eosin and Resorcin-Fuchsin. Samples processed with formalin and the H&E staining underlined various artifacts; vice versa, the Sandison solution associated with Resorcin-Fuchsin staining demonstrated the compression of the elastic fibres, with focal positivity in hanged corpses, and diffuse positivity in the entire hypostatic leaning. Sandison's rehydrating solution and Resorcin-Fuchsin staining exalt the supposed application of an asphyctic mean even on putrefied corpses in those cases burdened by a diagnostic doubt.

In pathological-forensic literature blast injuries are usually studied in relation to suicides, homicides during terroristic attacks and accidental events on the workplace, while the finding of these kinds of fatalities during misdoing is a recent occurrence. The Authors describe the peculiar case of the accidental death of the member of a criminal mob specialized in thefts to ATMs using acetylene tanks. The victim, having poured out the acetylene with the intent of triggering an explosion, was involved in the blast of the tank, deceasing immediately. This unusual manner of death highlights the danger of room saturation with acetylene, a criminal technique becoming more and more popular in the last years.

Introduction: In a previous work, we wanted to evaluate if the histochemical determination of lead in Gunshot Residues (GSR) on firearm wounds could be misled due to possible environmental contamination produced by heavy metals and, in particular, by lead. The Sodium Rhodizonate test and its confirmation test with 5% HCl Sodium Rhodizonate resulted to be negative and therefore we wanted to verify if these techniques were sensible enough in order to evaluate this element. We have assessed, on these same samples, a more sensitive technique, as inductive coupled plasma mass spectrometry (ICP-MS) is. This technique is able to detect elements in solution at concentrations as low as 10 -15 gL -1 ., Materials and Methods: Skin samples taken from two groups of victims, whose cause of death was not related to gunshot wounds were analyzed using ICP-MS: group A included 25 corpses found in open spaces after a long time; group B included 16 corpses exhumed after a period of 11 years. As a positive control group we used skin samples from two subjects that had died due to firearm wounds: as a negative control group we used three different types of plain paraffin slides without included biological material., Results: At the analysis by ICP-MS, the evaluation of the samples belonging to groups A, B and for the negative control groups resulted to be negative for traces of lead (Pb), barium (Ba) and antimony (Sb). On the other hand, high concentrations of GSR could be found in the positive control group were victims died for firearm wounds., Conclusions: On these basis, we can state that environmental Pb does not contaminate cadavers exposed to open air nor those buried in soil, as confirmed using to ICP-MS technique. Sodium Rhodizonate and 5% HCl Sodium Rhodizonate confirmation test have therefore a high sensitivity, highlighting GSRs, for the diagnosis of death caused by firearm wounds., (Copyright © 2018 Elsevier B.V. All rights reserved.)

The morphological analysis of stab wounds may often not be accurate enough to link it with the type of wounding weapon, but a further evaluation may be performed with the search for metallic residues left during the contact between the instrument and the skin. In this study, Scanning Electron Microscopy-Energy Dispersive Spectroscopy (SEM-EDS) was applied to the study of cadaveric stab wounds performed with kitchen knives composed of iron, chromium and nickel, in order to verify the presence of metallic residues on the wound's edge. Two groups of 10 corpses were selected: group A, including victims of stab wounds and a control group B (died of natural causes). Samplings were performed on the lesions and in intact areas of group A, whereas in group B sampling were performed in non-exposed intact skin. Samples were then analysed with optical microscopy and SEM-EDS. In group A, optical microscopic analysis showed the presence of vital haemorrhagic infiltration, while SEM-EDS showed evidence of microscopic metal traces, isolated or clustered, consisting of iron, chromium and nickel. Moreover, in two cases organic residues of calcium and phosphate were detected, as a probable sign of bone lesion. Control samples (group A in intact areas and group B), were negative for the search of exogenous material to optical microscopy and SEM-EDS. The results show the utility and possible application of the SEM-EDS in theidentification of metallic residues from sharp weapons on the skin., (Copyright © 2017 The Chartered Society of Forensic Sciences. Published by Elsevier B.V. All rights reserved.)

The authors applied scanning electron microscopy with energy-dispersive x-ray spectrometry to the furrow derived from hanging means. The study was conducted with the purpose to detect possible extraneous microtraces, deriving from the ligature, that could have had an interaction with the cutaneous biological matrix, thanks to a transfert mechanism, in the proximities of the lesion.Fifteen cutaneous samples of the furrow and an equal number of fragments of graphite tape, directly positioned on the lesion produced by the ligature mean and used as a "conductor" of possible traces, were analyzed using scanning electron microscopy with energy-dispersive x-ray spectrometry.The research of microscopic traces on the furrow using this technique highlights extraneous traces leading to 3 main categories: natural fabrics, and synthetic and metallic materials, excluding possible environmental pollutants. The analysis, run on 7 hanging deaths, made available by the judicial authority, found a morphological and compositional compatibility with the traces found on the cutaneous furrow produced during hanging.The technique used in this study is innovative in the pathological-forensic field, and can be considered useful in clarifying and studying this typology of asphyxia leading to a specific ligature material, when missing, or attributing the cause of death to hanging when the furrow is not macroscopically obvious.