Pilot Application of SEM/EDX Analysis on Suspected Cigarette Burns in a Forensic Autopsy Case of Child Abuse.

ABSTRACT: Cigarette burn lesions present forensic scenarios that are often difficult to investigate, both from a morphological diagnostic point of view and with regard to the mode of infliction, especially if the victim is unable to speak or has died. Although there may be the suspicion for a lesion to be produced by a lit cigarette, to date one can only rely on the morphological aspects that characterize it, and there is a lack of tools to reach the most evidence-based diagnosis possible. This limitation arose when managing a forensic autopsy case of possible child abuse that resulted in the death of the child, characterized by the presence of 3 suspicious cigarette burn lesions. We therefore decided to perform scanning electron microscopy/energy-dispersive x-ray (SEM/EDX) spectrometry analysis on these lesions and on the cigarette butt found at the crime scene. At the same time, SEM/EDX was applied to the analysis of an unlit cigarette in its entirety (obtained from the same source package as the cigarette butt), a positive control skin sample with an iatrogenic cigarette burn injury, and a negative control skin sample. Among the various compounds highlighted on compositional analysis, only sulfuric anhydride (SO 3 ) and phosphoric anhydride (P 2 O 5 ) showed a highly significant distribution pattern by being found in the autopsy samples, the cigarette butt, the tobacco of the unlit cigarette, and the positive skin control. Considering this, cigarette burns appear to follow Locard's principle as well, and similarly to other lesions, SEM/EDX allowed the diagnosis of cigarette burn lesions, already suspected morphologically, to be corroborated. Therefore, SEM/EDX is confirmed as a helpful tool in forensic pathology investigations.

Correlation of soft tissue and skeletal injuries in cases of violent death: A retrospective study of autopsy cases for forensic anthropology.

In forensic pathology, deaths due to mechanical injuries (blunt, sharp, and gunshot) require an autopsy in order to determine the cause and manner of death through a detailed examination of viscera, soft tissues, and the skeletal system. Sometimes, such as in cases of severe putrefaction, only the skeleton is observable. In such cases, the absence of bony defects does not mean that a trauma-related death should not be considered: yet, how often will a lethal mechanical death produce a defect in the skeletal system? The present study involved a retrospective review of autopsy reports to investigate the frequencies of soft tissue and related skeletal injuries in cases of violent deaths. A total of 200 autopsy reports for cases of four mechanisms of injury including sharp force trauma (n = 50), gunshot (n = 50), blunt force trauma (n = 50), and mechanical asphyxia (n = 50) were examined. Manner of death, localization of bone fractures, and correspondence between external injuries and bone fractures were reported. Frequencies of bone fractures and degree of correspondence with soft tissue injuries for each trauma group were evaluated. The result showed that lethal blunt trauma always caused bone fractures; 94.0% of violent deaths by gunshot involved bones; 48.0% of sharp force soft tissue injuries involved the bone tissue; and hyoid bone fractures were observed in 20.0% of cases of mechanical asphyxia.

Two peculiar cases of cranial fractures running through craniotomy burr holes: may this be a kind of "exception" to the Puppe rule.

A correct assessment on the position, path, and direction of fracture lines is crucial when the sequence of different injuries on the skull has to be ascertained. In this context, the so-called Puppe's rule on intersecting fracture lines has always been considered a cornerstone of such an investigation. However, there is one factor that has never been previously considered: how do fracture lines behave when they reach the edges of an old and remodeled hole from a previous craniotomy? Two peculiar cases are presented of subjects undergoing cranial fractures due to blunt force trauma (case 1) and gunshot (case 2). Both previously underwent neurosurgical operations with persistence of the burr holes produced by the craniotomy drill (15 and 20 years before death). What was arguable, according to Puppe's rule, was that the fracture lines, when at the edge of the craniotomy hole, stopped. However, what has been detected was different than what expected: fracture lines continued exactly in the opposite direction, as though they were "skipping" the hole, following the same direction and the same axis and stopping a few centimeters over on the opposite side of the craniotomy hole. Puppe's rule has never been refuted, but these cases are the closest to an exception ever seen in forensic anthropology.

The Lethal Cutting: An Unexpected Cause of Death-A Methomyl Acute Intoxication.

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.

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.

Cutaneous wound metallic residues pattern analysis with SEM-EDX in a forensic case of overkill.

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 overkill.

Thallium toxicity due to audultered infusion with thallium sulfate in eight members belonging to the same family nucleus: Autopsy findings and ICP-MS analysis (inductively coupled plasma mass spectrometry) in a triple homicide.

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.

Extensive study on electrocution at the Bureau of Legal Medicine of Milan (1993-2017): Determination of the current mark with scanning electron microscope/energy-dispersive X-ray analysis on paraffin-embedded samples.

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.

Technical note: A histochemical approach in diagnosing hanging mechanical asphyxia on cadavers undergoing advanced putrefactive phenomena.

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.

The comparative performance of PMI estimation in skeletal remains by three methods (C-14, luminol test and OHI): analysis of 20 cases.

When estimating post-mortem interval (PMI) in forensic anthropology, the only method able to give an unambiguous result is the analysis of C-14, although the procedure is expensive. Other methods, such as luminol tests and histological analysis, can be performed as preliminary investigations and may allow the operators to gain a preliminary indication concerning PMI, but they lack scientific verification, although luminol testing has been somewhat more accredited in the past few years. Such methods in fact may provide some help as they are inexpensive and can give a fast response, especially in the phase of preliminary investigations. In this study, 20 court cases of human skeletonized remains were dated by the C-14 method. For two cases, results were chronologically set after the 1950s; for one case, the analysis was not possible technically. The remaining 17 cases showed an archaeological or historical collocation. The same bone samples were also screened with histological examination and with the luminol test. Results showed that only four cases gave a positivity to luminol and a high Oxford Histology Index (OHI) score at the same time: among these, two cases were dated as recent by the radiocarbon analysis. Thus, only two false-positive results were given by the combination of these methods and no false negatives. Thus, the combination of two qualitative methods (luminol test and microscopic analysis) may represent a promising solution to cases where many fragments need to be quickly tested.

Analysis of metallic medical devices after cremation: The importance in identification.

The recovery of a charred cadaver raises many issues concerning personal identification; the presence of prosthetic materials may provide very important and decisive information. Who is involved in the recovery of a charred body or of burnt human fragments, should therefore be able to recognize medical devices even if modified by fire effects. Metallic residues (585kg) that came from 2785 cremations were studied. Medical devices were then divided by type and material in order to esteem the representativeness of each typology. The study illustrates the great presence of metal medical devices that could be of great help in identifying bodies and underlines that metallic medical devices types and morphology should be known by forensic practitioner involved in identification cases and that this kind of material can still be identified by physician and dentists, even if exposed to very high temperatures.

The medico-legal observation of an aggressive urogenital fibromatosis with isolated development not related to any traumatic event.

Desmoid tumor is a fibroproliferative neoplasm with an intermediate malignancy and it can be localized in every bodily district: some locations are considered exceptional, like the urogenital localization. The Author point out a rare case of giant idiopathic scrotal fibromatosis that was found during an autopsy. A widower, that lived alone in poor hygienic conditions, was found dead in his house. The Judicial Authority ordered the autopsy, that was performed two days later at the Medico-Legal Section of Milan University. External examinations revealed only the considerable dimension of the scrotum (cm 24 × 41). The cause of death was fixed in a cardiac tamponade due to a natural heart laceration localized in correspondence of a transmural infarction. The toxicological exam resulted negative, while the histopathological and immunohistochemical analysis qualify the scrotal mass as a desmoids tumor. Due to the absence of predisposing conditions and of fibroproliferative infiltration in bladder and retroperitoneal space, the neoplasm was configured as an idiopathic desmoid tumor. The presented case gives the reason for the discussion concerning medico-legal aspects that are typical of rare neoplasms.

Two decades of adolescent suicides assessed at Milan University's medicolegal unit: Epidemiology, forensic pathology and psychopathology.

AIM: to analyze the cases of suicide committed by adolescents in and around the city of Milan over a 20-year period (1993-2012). MATERIALS AND METHODS: cases of suicide involving individuals between 10 and 19 years of age were drawn from 20,757 autopsies performed by the Medico-legal Unit at Milan University. Seventy-eight cases were considered (20 females and 58 males, with the ratio of 1:2.9), and their clinical and circumstantial histories, epidemiology, forensic pathology and psychopathological issues were analyzed. RESULT: Adolescents were involved in 2.23% of all suicides committed in Milan during the period examined. The "mean" victim is a male without psychiatric disease, aged between 16 and 19, that commits suicide outdoors. CONCLUSIONS: In the majority of cases, there were no premonitory signs, nor any particular contingent or remote reason that might explain these violent deaths, which therefore remain "impulsive".

Characterisation of the weapon used in a patricide by SEM/EDS analysis of a microscopic trace from the object.

This article presents a case of patricide. The murder was characterised by multiple blunt traumas and asphyxia. A mass of contused wounds was localised to the head and neck, and included the complete avulsion of the left eye (by an unknown tool), which was recovered near the cadaver. This case is of interest due to the possibility of identifying microscopic traces of the object that was used for the homicide by examining the skin margins around the ocular injury. The analysis was conducted using scanning electron microscopy combined with energy dispersive X-ray analysis (SEM/EDX). Analysis of the skin margins allowed microscopic inorganic traces to be detected, which were identified as ceramic material. This result focused the attention of the investigation on a small fish-shaped statue that had been previously found by the police when examining the crime scene. The use of SEM/EDX was therefore essential in determining a match between the microscopic traces detected on the perilesional skin and the composition of the statue. This led to the suspicion that the statue was the murder weapon.

Detectability and medico-legal value of the gunshot residues in the intracorporeal channel.

The application of the histochemical stain of sodium rhodizonate to the entrance wound for the detection of the lead (Pb) residues coming from the gunshot may be affected by false positive cases due to the contamination of the environmental Pb. The aim of the Authors is to histochemically search the Pb of GSR in a region which should be more protected by the contamination: the intracorporeal channel. Two hundreds and eighteen serial histological specimens of the intracorporeal channels coming from 25 subjects (dead due to gunshots and being autopsied at the Section of Legal Medicine of the Milan University, in the years 2013-2014) were stained with the sodium rhodizonate and sodium rhodizonate in acid environment (HCl 5%), and then observed by the microscope. The sodium rhodizonate showed a positivity for the Pb residues in the intracorporeal channel, with the detection of the particles within the first 2 cm beyond the entrance wound in 6 cases over the total number of 25 (24%). Victims were characterized by common features: short-barreled weapon; contact shots or short-distance shots; involvement of regions that were not covered by clothing; preservation of the microscopic structure of organs interested by the intracorporeal channel. The searching of GSR in the intracorporeal channel, even in conditions securing a high sensitivity, could represent an important test for the discrimination between an environmental contamination of Pb and the presence of Pb residues by GSR: once confirmed the presence of GSR in the intracorporeal channel by the histochemical analysis, the diagnostic process should require the application of the SEM-EDX for the confirmation of the results. Although not yet studied, this combination could be applied to cadavers exposed to the environment, with advanced post-mortal phenomena permitting at least the suspects of the existence of gunshot wounds at the macroscopic autopsy evaluation. Indeed, in some cases, the putrefaction is so advanced that no suspect of gunshot injuries could be derived from the soft tissue.

Suicide drowning in the non-coastal territory of Milan.

INTRODUCTION: A cadaver in water sets a relevant issue for the forensic pathologist, concerning the differential diagnosis between homicide, suicide, accidental event, and natural death in water. In the present study, the drowning suicides in a non-coastal territory are analyzed, discussing the main diagnostic difficulties, the preventive intervention, and the strategies for outlining the profile of a "mean victim" that is at risk of committing suicide by drowning. METHODS: The cases of drowning suicide were extrapolated by the database of the 21,472 autopsies performed at the Civic Morgue of Milan, in the period between 1993 and 2013. RESULTS: Drowning represents the fifth cause of suicide registered in Milan for the study period, with 139 cases (3.8% of all suicides). Victims were mainly elderly males (61-80 years old) with a history of psychiatric diseases in pharmacological therapy; they committed suicide mainly during summer, in the rivers. The diatoms test was positive in 82 cases (59%) and highlighted typical taxa of fresh water. Alcohol and illicit drugs were not noticed, at high levels. CONCLUSION: Drowning suicide occurs even in a non-coastal territory, far from the sea, if there is an easy access to the water: specifically, many natural and artificial waterways in the Milan district represent a high-risk factor predisposing the drowning suicide.

Transphobic Murders in Italy: An Overview of Homicides in Milan (Italy) in the Past Two Decades (1993-2012).

In Europe, 71 murders resulting in the death of transgendered persons were reported between 2008 and 2013, 20 of which perpetrated in Italy, the second highest rate in Europe after Turkey. We retrospectively analyzed the homicides of transgender people recorded at the Medicolegal Bureau in Milan from January 1993 to December 2012. First we considered the sociodemographic data of 20 victims and the circumstantial details of their deaths, then we examined the data related to the cause of death from the autopsy reports. Our data show that victims are mostly immigrants, biological males presenting with a feminine attire and with varying degrees of feminization. The large majority of the victims were sex workers from South America. As for murderers, they were unknown in 7 cases (35%); all the 13 murderers identified were males, aged between 17 and 63 (M age = 31 years). In 38% of the cases, the murderer was the victim's current or former partner. For half of the homicides, it was possible to identify at least one primary indicator of lesbian, gay, bisexual, and transgender (LGBT) hate crime. Our findings call for the need to make explicit in Italian legislation that a crime perpetrated on the ground of sexual orientation and gender identity constitutes a hate crime.

An unusual case of homicide with a crossbow and a hunting knife.

Traumatic lethal injuries caused by crossbows are a rarity in forensic pathology. They occur as accidents, suicide and, more rarely, as murders. We report a case of murder received at the Institute of Legal Medicine in Milan carried out by two weapons - a crossbow and a hunting knife - that resulted in multiple thoraco-abdominal wounds. The case is also rare because of the unique circumstances involving the victim - a local service psychiatrist - and the perpetrator - a former medical specialist in psychiatry, suffering from a delusional psychotic disorder. The crossbow was the decisive instrument in the realization of the murder; it was chosen with premeditation for its high destructive capacity and the fact that it was easily obtainable, and it was used with darts that were specially modified to increase its harmful capacity. The presented case, as well as providing an opportunity to highlight the damaging effects of a rarely used weapon, stresses the problems inherent with access to tools such as the crossbow that are used in sport, and which are potentially dangerous but easily accessible, even to those with serious mental disorders, rendering them legally unfit for using firearms.

Preservation of histological structure of cells in human skin presenting mummification and corification processes by Sandison's rehydrating solution.

INTRODUCTION: To overcome the difficulties of construction and interpretation of microscopic material from corpses presenting mummification and corification processes, a variety of techniques and tricks are used: in this research the results of applying the Sandison's rehydrating solution are presented, generally used in archeological field on Egyptian mummies of different ages, in human cadaveric material in an advanced state of decomposition. MATERIALS AND METHODS: Nineteen skin specimens were taken from corpses presenting corification and mummification processes, discovered in a time ranging between one and four months and exhumed after 11 years. Each biological sample was divided into two parts: one, directly fixed in buffered formalin 10%; the other, preliminarily treated with the Sandison's rehydrating solution and, therefore, post-fixed in 10% buffered formalin. All samples were then carried out the routine histological preparation, and the sections were stained by hematoxylin-eosin and by other histochemical stains. RESULTS: Under the microscope, the samples placed directly into formalin, showed marked structural changes of the various components, while those previously rehydrated with the Sandison's rehydrating solution allowed the clear recognition of different structures. CONCLUSIONS: The use of the Sandison's rehydrating solution on skin samples presenting corification and mummification processes, preserving significantly its general setting, stands as an indispensable procedure in the study of such cases.

Applicability of cranial models in urethane resin and foam as a substitute for bone: are synthetic materials reliable?

As literature is poor in functional synthetic cranial models, in this study, synthetic handmade models of cranial vaults were produced in two different materials (a urethane resin and a self-hardening foam), from multiple bone specimens (eight original cranial vaults: four human and four swine), in order to test their resemblance to bone structure in behavior, during fracture formation. All the vaults were mechanically tested with a 2-kg impact weight and filmed with a high-speed camera. Fracture patterns were homogeneous in all swine vaults and heterogeneous in human vaults, with resin fractures more similar to bone fractures. Mean fracture latency time extrapolated by videos were of 0.75 msec (bone), 1.5 msec (resin), 5.12 msec (foam) for human vaults and of 0.625 msec (bone), 1.87 msec (resin), 3.75 msec (foam) for swine vaults. These data showed that resin models are more similar to bone than foam reproductions, but that synthetic material may behave quite differently from bone as concerns fracture latency times.