Sudden Death due to Diseases of the Adrenal Glands and Paraganglia

Coroners and pathologists commonly evaluate unexpected deaths due to diseases of the adrenal glands and paraganglia, which are, unfortunately, not rare in their totality. Although cardiac causes are the main cause of sudden death, endocrine conditions can produce sudden, unexpected deaths that need further investigation, especially in younger patients. This chapter focuses on the issue of sudden death due to diseases involving adrenal glands and paraganglia. The main causes of sudden death due to adrenal gland pathology will be examined, paying particular attention to the pathophysiology of sudden death, macroscopic and microscopic characteristics and their correlation with clinical features. These issues are of great interest, especially considering the clinical impact of sudden death and its rarity among patients with adrenal gland diseases. The forensic pathologist’s examination is extremely important in determining the cause of death and findings not clinically observable and can contribute to the improvement of the clinical and surgical approach in treating such patients

Diagnosis of sudden cardiac death due to early myocardial ischemia: An ultrastructural and immunohistochemical study

The aim of this post-mortem ultrastructural and immunohistochemical study is to explore the characteristics of acute myocardial ischemia in the context of sudden death, using the combination of two different methods, both more insightful than ordinary histology. Transmission electron microscope and immunohistochemistry, in addition to the traditional histology, were applied to study human heart specimens collected during forensic autopsies. The whole series was sub-grouped into cases (n=17) and controls (n=10). The control group consisted of unnatural death with a short agonal period (immediately lethal injuries). Heart samples of the two cohorts of subjects were prepared for electron microscopy. On the other hand, each specimen, formalin fixed and paraffin embedded, was stained with haematoxylin and eosin and immunoreacted with the following primary antibodies: antiFibronectin, antiConnexin-43, anti npCx43 (dephosphorylated form of Connexin43), antiZonula occludens-1. Immunopositivity of each marker in the myocardium was semi-quantitatively graded. Electron microscopy revealed a number of interesting differences between acute myocardial ischemia and controls, regarding the morphology of nucleus, mitochondria and intercellular junctions. By immunohistochemistry, fibronectin was found to be markedly increased in the extracellular matrix of the acute myocardial ischemia cases, with a remarkable difference in respect of controls. Connexin 43 staining disclosed a slightly increase in the cytoplasm of acute myocardial ischemia cases with respect to the controls, whereas no relevant differences were seen between cases and controls at intercellular junctions. Dephosphorylated form of Cx43 showed an evident difference of staining in cases compared to controls and overall this difference more evident in the cytoplasm. Zonula occludens 1, described as an important marker for functional modification of cardiac muscle fibers, resulted negative or very weak in the vast majority of both cases and controls. The present study attempts to simultaneously apply electron microscopy and immunohistochemistry, in order to figure out the morphological changes that might lead to pathological processes underlying the sudden, unexpected death due to acute myocardial ischemia, and consequently to find useful diagnostic markers of very early ischemic injury. Both methods showed significant differences between acute myocardial ischemia and controls, regarding, overall nuclei, mitochondria, and intercellular junctions.

Asbestos exposure and malignant mesothelioma: the role of inorganic fiber burden and disruption of iron homeostasis in lung microenvironment. A postmortem study on human lungs

Asbestos-related diseases still represents a major public health problem all over the world. Among them, malignant mesothelioma (MM) is a highly aggressive, poor-prognosis cancer, arising from the serosal lining of pleura, pericardium and peritoneum, triggered by asbestos exposure. Asbestos is the collective name of six kinds of naturally occurring minerals, namely chrysotile (the only serpentine asbestos), and the amphiboles crocidolite, amosite, tremolite asbestos, actinolite asbestos, anthophyllite asbestos. The response of human lungs to asbestos inhalation and the molecular mechanisms which lead to MM development several decades after exposure are still largely unknown. One of the most debated issues is the formation of asbestos bodies, that are asbestos fibers covered by an iron-rich amorphous substance. Literature data suggest the key role of iron metabolism in the coating process leading to the formation of asbestos bodies, that has been regarded as both protective and harmful. This study aims to understand more about the reaction of the human organisms to asbestos inhalation and the individual susceptibility to MM. First, the lung inorganic fiber burden has been characterized in lungs of individuals who were previously exposed to asbestos (occupationally or environmentally) using electron scanning microscopy with energy dispersive spectroscopy (SEM-EDS). All the subjects used to work at an asbestos-cement industry, which was active between 1932 and 1993 in Broni, a small town in Lombardy. Unexpectedly, a significantly lower concentration of asbestos fibers has been found in MM compared to asbestosis patients. Chrysotile was not detected at all in any of the examined samples, despite it was largely used at the plant, suggesting a complete clearance of this type of asbestos from lungs. Crocidolite was the most represented asbestos, followed by amosite, tremolite/actinolite asbestos and anthophyllite asbestos, consistently with the data about the industry production. The ratio between asbestos fibers and asbestos bodies was widely different from subject to subject. Based on the well-known role of iron in asbestos-induced pulmonary toxicity, the second part of the study investigated the frequency of a group of single nucleotide polymorphisms (SNPs) in genes involved in iron homeostasis in individuals who died from MM compared to controls. Despite the successful DNA extraction from formalin-fixed paraffin-embedded samples (FFPE), we failed to identify any genotype associated with a protective or predisposing effect in relation to MM development as a consequence of asbestos exposure. Finally, on the basis of the established role of BAP1 in MM pathogenesis and its association with ferroptosis impairment observed in various kinds of cancers, the expression of BAP1, transferrin receptor 1 (TRF1), ferritin heavy chain 1 (FTH1) and ferroportin (FPN) has been investigated using immunohistochemistry and rtPCR, finding that asbestos affects the expression of the mentioned proteins in lungs differently in MM patients compared to subjects exposed to asbestos but died of other causes. These findings suggest that a different biological response to asbestos inhalation and to the consequent iron overload in lungs may play an important role in cancer initiation. The formation of asbestos bodies appears to be a key mechanism in the formation of a pro-neoplastic microenvironment, as well as ferroptosis impairment. These results are important from a prevention point of view, as iron metabolism, as well as the consequent oxidative stress, chronic inflammation and cancerogenic stimuli might be targets for therapeutic strategies aiming to delay or prevent MM onset in individuals previously exposed to asbestos. Moreover, knowing the mechanism that can make an individual vulnerable to asbestos can be of crucial importance for prevention

GENETIC INDIVIDUAL IDENTIFICATION FROM DRIED URINE SPOTS (DUS): A COMPLEMENTARY TOOL TO DRUG MONITORING AND ANTI DOPING TESTING

8: The collection of liquid biological matrices onto paper cards (Dried Matrix Spots - DMS) is becoming an alternative sampling strategy. The stability over time of molecules of interest for therapeutic, sport drug monitoring and forensic toxicology on DMS has been recently investigated representing a reliable alternative to conventional analytical techniques. When a tampering of a urine sample in drug monitoring or doping control cases is suspected, it could be relevant to know whether genetic profiles useful for individual identification could be generated from urine samples spotted onto paper (Dried Urine Spot - DUS). To understand the influence of sex, storage conditions and time on the quality and quantity of the DNA, five female and ten male urine samples were dispensed onto Whatman 903 paper and sampled after different storage conditions over time, from 1 to 12 weeks. Direct PCR was performed starting from 2 mm punches collected from each spot amplifying a panel of markers useful for individual identification. The female DUS stored in different conditions produced genetic profiles fully matching the reference samples. The same result was obtained for the male DUS but using urine 30X concentrated by centrifugation instead of the original samples. Our data show that this approach is valid for genetic individual identification of urine samples spotted onto paper cards up to 12 weeks after deposition and could be easily incorporated in anti-doping or drug screening protocols to help on the suspicion of evidence tampering or to solve questions on the reliability of samples collection.nonemixedGrignani, Pierangela; Manfredi, Alessandro; Monti, Maria Cristina; Moretti, Matteo; Morini, Luca; Visonà, Silvia Damiana; Fattorini, Paolo; Previderè, CarloGrignani, Pierangela; Manfredi, Alessandro; Monti, Maria Cristina; Moretti, Matteo; Morini, Luca; Visonà, Silvia Damiana; Fattorini, Paolo; Previderè, Carl

Death of a seven-month-old child in a washing machine: a case report

The authors present a case which brings out a unique modality of child homicide by placing the baby in a washing machine and turning it on. The murder was perpetrated by the babyâs mother, who suffered from a serious depressive disorder. A postmortem RX and then a forensic autopsy were performed, followed by histologic examinations and toxicology. On the basis of the results of the autopsy, as well as the histology and the negative toxicological data, the cause of death was identified as acute asphyxia. This diagnosis was rendered in light of the absence of other causes of death, as well as the presence of typical signs of asphyxia, such as epicardial and pleural petechiae and, above all, the microscopic examinations, which pointed out a massive acute pulmonary emphysema. Regarding the cause of the asphyxia, at least two mechanisms can be identified: drowning and smothering. In addition, the histology of the brain revealed some findings that can be regarded as a consequence of the barotrauma due to the centrifugal force applied by the rotating drum of the washing machine. Another remarkable aspect is that we are dealing with a mentally-ill assailant. In fact, the babyâs mother, after a psychiatric examination, was confirmed to be suffering from a mental illnessâa severe depressive disorderâand so she was adjudicated not-guilty-by-reason-of-insanity. This case warrants attention because of its uniqueness and complexity and, above all, its usefulness in the understanding of the pathophysiology of this particular manner of death