Corpses against science death: new Italian law titled 'Rules relating to the directives of one's body and tissues post-mortem for study, training, and scientific research purposes'.

The new Italian Law no. 10 of 2020, titled 'Rules relating to the directives of one's body and tissues post-mortem for study, training, and scientific research purposes' (10 February, 2020), has opened the doors to a new and clearer regimentation on body donation to science, according to the procedure already introduced by Law no. 219/2017 on the 'Rules relating to informed consent and advance directives of treatment'. The authors exposed some concerning regarding the application of the law, such as donation mistrust, also linked to our cultural heritage; the incomplete protection of minor and incompetent people; the complexity of an adequate information, which could undermine the informed consent; and finally, the fees overload for research institutes, with already exiguous budgets. Next, the authors analysed some positive aspects of the law and in primis the central role of corpses in scientifical knowledge and medical training. Moreover, a simple mechanism based on a conscious and pre-stated consent, registered in a database that follows European data protection Regulation, is discussed. Further, the procedure of the restitution of the bodies to their families, which represents a way to guarantee the relatives' commemoration, is also elucidated. In conclusion, the authors proposed a format of informed consent, to try a direct application of the law, thereby fastening the process of standardisation and centralisation of all the living wills. The authors optimistically investigated the opportunities offered by the law and encouraged a conscientious participation by citizens.

[Acroosteolysis in PVC autoclave cleaners: history of an occupational disease]. / Nascita, sviluppo e scomparsa di una malattia professionale: la acroosteolisi dei pulitori manuali di autoclavi nella produzione di PVC.

OBJECTIVES: This paper examines the history of an occupational disease which has now disappeared: acroosteolysis of manual tank cleaners in the production of polyvinyl chloride (PVC), which is a rare disease characterized by destructive alterations of the distal phalanges of the hands. METHODS: All the available literature on this disease was examined. The history of acroosteolysis was studied within the general framework of the history of the discovery of adverse health effects of exposure to vinyl chloride, and this history was studied up to the end of the 1960's. RESULTS: The disease was observed for the first time in mid-1963 in Belgium (Jemeppe) in a chemical plant operated by Solvay, and affected two workers whose job was the manual cleaning of vessels used for the polymerization of vinyl chloride; similar cases occurred in almost all PVC production plants all over the world, but not in the plants where the main activity was the production of vinyl chloride monomer (VCM). Little more than one hundred cases are described in the scientific literature, and this number increases by a few dozen if we consider known but unpublished cases. These figures confirm the rarity of the disease, which peaked at the end of the 1960's and disappeared during the 1970's, probably due to the complete elimination of manual reactor cleaning. Observation of the disease lasted no more than fifteen years and the disease was not replicated in experimental conditions on animals. DISCUSSION: The disease was clinically characterized, had a short latency (from several months to several years), was rare and unequivocally linked to the manual cleaning of PVC polymerization tanks. However many questions still remain open: the period when the disease first appeared (many years after the start of PVC production in the world), the etiology of the disease (the most accredited hypothesis considers three concomitant factors: a chemical factor--one of the many substances used during polymerization, and particularly vinyl chloride monomer, a physical factor--microtraumas of the fingers during manual cleaning, individual susceptibility), the pathogenetic mechanism (in particular: the role of skin, respiratory, or digestive system, as entrance door), a method (or test) to screen subjects potentially predisposed to the disease. In our view acroosteolysis of manual tank cleaners in PVC production is an occupational disease which is distinct from "vinyl chloride disease" as identified by Viola (1974).

Diagnosis of electrocution: The application of scanning electron microscope and energy-dispersive X-ray spectroscopy in five cases.

INTRODUCTION: Deaths from electricity, generally, do not have specific findings at the autopsy. The diagnosis is commonly based on the circumstances of the death and the morphologic findings, above all the current mark. Yet, the skin injury due to an electrocution and other kinds of thermal injuries often cannot be differentiated with certainty. Therefore, there is a great interest in finding specific markers of electrocution. The search for the metallization of the skin through Scanning Electron Microscope equipped with Energy Dispersive X-Ray Spectroscopy (EDS) probe is of special importance in order to achieve a definite diagnosis in case of suspected electrocution. MATERIALS AND METHODS: We selected five cases in which the electrocution was extremely likely considering the circumstances of the death. In each case a forensic autopsy was performed. Then, the skin specimens were stained with Hematoxylin Eosin and Perls. On the other hand, the skin lesions were examined with a scanning electron microscope equipped with EDS probe in order to evaluate the morphological ultrastructural features and the presence of deposits on the surface of the skin. RESULTS: The typical skin injury of the electrocution (current mark) were macroscopically detected in all of the cases. The microscopic examination of the skin lesions revealed the typical spherical vacuoles in the horny layer and, in the epidermis, the elongation of the cell nuclei as well as necrosis. Perls staining was negative in 4 out 6 cases. Ultrastructural morphology revealed the evident vacuolization of the horny layer, elongation of epidermic cells, coagulation of the elastic fibers. EDS-MICROANALYSIS: In the specimens collected from the site of contact with the conductor of case 1 and 2, the presence of the Kα peaks of iron was detected. In the corresponding specimens taken from cases 2, 4, 5 the microanalysis showed the Kα peaks of titanium. In case 3, titanium and carbon were found. CONCLUSIONS: In the suspicion of electrocution, the integrated use of different tools is recommended, including macroscopic observation, H&E staining, iron-specific staining, scanning electron microscopy and EDS microanalysis. Only the careful interpretation of the results provided by all these methods can allow the pathologist to correctly identify the cause of the death. Particularly, the present study suggests that the microanalysis (SEM-EDS) represents a very useful tool for the diagnosis of electrocution, allowing the detection and the identification of the metals embedded in the skin and their evaluation in the context of the ultrastructural morphology.

A case of sudden death after ultrasound-guided percutaneous alcohol injection of a paraganglioma mis-diagnosed as a peri-renal cyst.

Paragangliomas are cromaffin tumors arising from the neural crest cells of parasympathetic or sympathetic ganglia. They are known to be rare causes of sudden death. Here we present the autopsy findings, as well as microscopical and immunohistochemistry study, of a 48-years-old woman who died suddenly after a percutaneous alcohol injection of a peri-renal cyst previously diagnosed as a common complex cyst. She manifested a multiorgan failure, with acute heart failure, systemic and pulmonary vasoconstriction with hypoxia, metabolic acidosis (pH 6.974). It was therefore enacted resuscitation that was ineffective. The autopsy pointed out, close to the upper right renal pole, a cyst characterized by very friable walls and septa, with a thickness of approximately 0.5cm and containing about 75cm(3) of hemoserous fluid. Microscopically, through immunohistochemical examinations, the cyst showed the presence of chromaffin cells, containing enzymes involved in the synthesis of catecholamines, in particular noradrenalin. So, the cause of the death was ascertained in an multi-organ failure caused by a massive release of catecholamines (noradrenaline) from the cyst, identified post-mortem (on the basis of histologic and immunohistochemical examinations) in a noradrenalin-secreting paraganglioma, that remained silent until the cyst ablation.

Achalasia and sudden death: a case report.

A 23-year-old pregnant woman was found dead at home. The anamnesis was completely negative, except for a slight dysphagia. The most important autopsy finding was a remarkable previously undiagnosed, megaesophagus. Conjunctival and pleural petechiae were also observed. The most likely etiopathogenetic factors are considered and discussed.

Electron spectroscopic imaging of secretory granules in human eccrine sweat glands.

A study of human eccrine sweat glands by electron spectroscopic imaging has been performed. A high nitrogen content in secretory granules of the dark mucous cells has been found while mucous cells in the digestive or respiratory systems did not contain nitrogen detectable in our experimental conditions. High concentrations of nitrogen were found in zymogenic granules and in the core of the granules in the eosinophilic leucocytes. Our findings seem to suggest that the granules of the dark secretory cells are composed not only of mucoid substances but contain also high concentrations of proteins. The descriptive term "granulated cells" seems to be more correct than "mucous cells" to indicate these elements.

Parapharyngeal abscess and sudden death.

This case-report illustrates a sudden asphyctic death of a 68-year-old female caused by an inflammatory obstruction in the upper airway. The autopsy revealed a peritonsillar-retropharyngeal abscess. Possible explanations for this rapid and lethal progress of a tonsillar infection are discussed.

Observations on dental prostheses and restorations subjected to high temperatures: experimental studies to aid identification processes.

In large scale disasters associated with fire the damage caused by heat can make medico-legal identification of human remains difficult. Teeth, restorations and prostheses, all of which are resistant to even quite high temperatures can be used as aids in the identification process. In this project the behaviour and morphology of teeth and dental prostheses exposed to a range of high temperatures was studied. Healthy teeth, dental restorations and prostheses were placed in a furnace and heated at a rate of 30 degrees C/min and the effects of the predetermined temperatures 200, 400, 600, 800, 1000 and 1100 degrees C were examined by stereomicroscopy and scanning electron microscopy (SEM). Our observations show that some prostheses and restorative materials resist higher temperatures than theoretically predictable and that even when a restoration is lost because of detachment or change of state its ante-mortem presence can be confirmed and detected by both stereomicroscopic examination and SEM of the residual cavity. We further conclude that a reasonably reliable estimation of the temperature of exposure can be made from an analysis of the teeth and restorative materials.

Astrocytic reaction in long-term resuscitation.

The brain of 34 subjects who died after prolonged resuscitation (more than 20 days) were studied in order to evaluate the possibility of discriminating original cerebral injuries from changes caused by prolonged resuscitation. We focused our attention on the astrocytic reaction and its relationship to the associated lesions. Twelve of the 34 cases showed astrocytosis picture. Of these 5 were anisomorphic and 7 isomorphic. From the analysis of the various morphological findings, taking into account an individual's clinical history, we can conclude that isomorphic astrocytosis is a typical morphological reaction related to resuscitation duration and it is the result of a slow, gradual neuronal degeneration induced by chronic hypoxia. Anisomorphic astrocytosis, on the other hand, can be an indicator of acute and focal cerebral lesion.

Exposure to priority organochlorine contaminants in the Italian general population. Part 2: Fifteen priority polychlorinated biphenyl congeners in blood serum.

Concentrations of 36 polychlorinated biphenyl (PCB) congeners were measured in serum of 372 Italian residents of general population living in Novafeltria, Pavia, and Milan. Total PCB level differed significantly between these sites (p < 0.0001) with median concentrations of 836.50, 1354.57, and 2062.08 pmol/g lipid, respectively. However, there is no evidence for the difference in distribution of total PCB levels by genders. Total dioxin-like PCBs differed significantly (p < 0.0001) between the sites (median 109.78, 50.88, and 166.99 pmol/g lipid, respectively) and genders of Novafeltria and Pavia (p = 0.011 and 0.009, respectively). PCB 138, 153, 170, and 180 differed significantly between the places of residence (p < 0.0001) with higher values in Milan population. In the overall population, total PCB and PCB 138, 153, 156, 170, and 180 correlated positively with age (correlations range between 0.320 and 0.569, p < 0.0001). In Novafeltria, the correlations ranged between 0.545 and 0.670, and in Pavia, the correlations ranged between 0.516 and 0.666. In Milan, correlations with age range between 0.327 and 0.417 for total PCB and congeners 138, 153, and 180. With an exception of PCB 170, there was no evidence of significant difference in the distribution of most abundant PCB congeners and total PCB across the body mass index categories.

Exposure to priority organochlorine contaminants in the Italian general population. Part 1. Eight priority organochlorinated pesticides in blood serum.

Despite extensive use of organochlorinated pesticides (OCPs) such as dichlorodiphenyltrichloroethane (DDT) in Italy in the 1940s to 1970s, especially for public health control of malaria mosquitoes, information on their exposure levels among the general population is limited. These OCPs can be a source of health risk to human. A total of 137 blood samples were collected from residents of the general population of three Italian towns, Novafeltria, Pavia and Milan, to determine the levels of eight OCPs in blood serum. The concentrations of beta-hexachlorocyclohexane, hexachlorobenzene (HCB), 1,1-dichloro-2-(o-chlorophenyl)-2-(p-chlorophenyl)ethylene, 1,1-dichloro-2,2-bis(p-chlorophenyl)-ethylene (p,p'-DDE), 1,1-dichloro-2-(o-chlorophenyl)-2-(p-chlorophenyl)ethane, 1,1-dichloro-2,2-bis (4-chlorophenyl)ethane, 1,1,1-trichloro-2-(o-chlorophenyl)-2-(p-chlorophenyl)-ethane and 1,1,1-trichloro-2,2-bis(p-chlorophenyl) ethane were measured by gas chromatography-mass spectrometry. Variations in serum concentrations of OCPs with respect to place of residence, gender, age and body mass index (BMI) were evaluated by non-parametric tests. p,p'-DDE and HCB were the most abundant and major contributors of total OCP concentration. Their levels differed significantly between the three towns with a trend Milan > Novafeltria > Pavia (p < 0.0001). Females had significantly higher concentrations of HCB and p,p'-DDE than males in the overall population sample. HCB concentrations were significantly higher in females than in males of Milan (p = 0.029). We observed positive correlations of p,p'-DDE and HCB with age in Novafeltria subjects (r = 0.468, p = 0.004). Total OCP concentrations differed significantly across BMI categories (p = 0.018) in overall population. We have demonstrated a clear pattern of the main OCPs in a fairly large population. Generally, our study provides information on OCPs exposure among the Italian general population and provides indications for further investigations.

Two cases of accidental asphyxia by neck compression between bed bars.

Two cases of asphyxia due to compression of the neck between the side bars of a bed in elderly subjects affected by neuropsychiatric pathologies are presented. In both cases no lesions were found on the skin or in the anatomic structures of the neck. The absence of lesions made determining the cause of death difficult. Generic evidence of asphyxia (acute pulmonary emphysema and petechiae) allowed a diagnosis to be formulated. The discovery of the object that caused the death was possible only with information regarding the circumstances and inspection of the scenes of the deaths.

Morphology of sweat glands in determining time of death.

This study demonstrates post-mortem autolytic alterations in the skin at cellular and subcellular levels and identifies parameters which may assist in determining the time of death in the first few hours post-mortem. Serial skin samples from the ventral surface of the arm were taken at intervals of 3, 6, 9 and 12 h after death in 29 subjects of various ages, with no signs of skin disease; causes of death were various. Three types of tests were performed: cytochemical (hematoxylin-eosin and alcian-PAS), immunohistochemical (S-100, CEA, Cytokeratin, ASM) and ultrastructural (electron microscopy). Electron microscopy proved useful for identifying transformations which were found to be specific for each chronological step considered: reduction of intracellular glycogen in clear cells and reduction of secretory granules in dark cells are typical signs of the first stage (3 h) after death; mitochondrial dilatation and rarefaction of cristae in clear and dark cells are typical of the second stage (6 h); rarefaction of microvilli in dark and clear cells is a sign of the last stage (12 h). Cytochemistry and immunohistochemistry supply useful information--not for all the chronological stage considered here, but for individual phases (3 h for hematoxylin-eosin and 6 h for alcian-PAS). However, it is particularly important to use the results from all such techniques simultaneously, so that the question of the exact time of death within the first 12 h post-mortem may be more accurately answered.

Immunohistochemical evidence suggests intrinsic regulatory activity of human eccrine sweat glands.

Immunohistochemistry of normal eccrine sweat glands was performed on paraffin sections of human skin. Immunoreactivity (ir) for neuron specific enolase, S100 protein (S100), regulatory peptides, nitric oxide synthase type I (NOS-I) and choline-acetyltransferase (ChAT) was found in small nerve bundles close to sweat glands. In the glands, secretory cells were labelled with anticytokeratin antibody. Using antibodies to S100, calcitonin gene-related peptide (CGRP) and substance P (SP) a specific distribution pattern was found in secretory cells. Granulated (dark) and parietal (clear) cells were immunopositive for CGRP, and S100 and SP, respectively. Immunoreactivity was diffuse in the cytoplasm for CGRP and S100, and peripheral for SP. Myoepithelial cells were not labelled. Electron microscopy revealed electron dense granules, probably containing peptide, in granulated cells. Using antibodies to NOS-I and ChAT, ir was exclusively found in myoepithelial cells. Immunoreactivity for the atrial natriuretic peptide was absent in sweat glands. These results provide evidence for the presence of both regulatory peptides involved in vasodilation and key enzymes for the synthesis of nitric oxide and acetylcholine in the secretory coil of human sweat glands. It is suggested that human sweat glands are capable of some intrinsic regulation in addition to that carried out by their nerve supply.

Regulation of inflammatory responses to Bordetella pertussis by N(G)-monomethyl-L-arginine in mice intranasally infected.

To investigate effect of MMLA, an inhibitor of nitric oxide (NO) production, on regulation of inflammatory responses to Bordetella pertussis infection, mice were infected intranasally, and treated with various concentrations of MMLA. Ten days after infection, mice treated with MMLA at dosage of 100 mg/kg, given intraperitoneally in a single dose or for 5 consecutive days, showed at histopathologic examination, a significant decrease of intensity of inflammation (scores, 0.6 +/- 0.2 and 0.9 +/- 0.5 respectively). A decrease of cellular accumulation of neutrophils and lymphocytes in the bronchoalveolar lavage (BAL) fluid was observed in infected mice treated with MMLA, especially at dosage of 10 mg/kg, given in a single dose intraperitoneally. In addition, BP-infected mice treated with MMLA (100 mg/kg, intraperitoneally) for 5 consecutive days showed higher mortality rate than untreated mice infected with B. pertussis, and the number of B. pertussis in lungs of mice treated with MMLA was significantly increased. However, MMLA treatment of infected mice had some effect on levels of IFN-gamma and nitrite/nitrate (end-stable products of NO) in the BAL fluid. This study indicates that NO may play a role either as microbiocidal agent or as a modulator of immune regulation, inasmuch as it may upregulate tissue inflammatory response to B. pertussis.

Allo-activated CD4+ and CD8+ lymphocyte subsets: new ultrastructural findings based on computer-assisted image analysis.

Primary allo-stimulation induces substantial and differentiated modifications in the development of cytoplasmic organelles of peripheral T-lymphocyte subsets. The present computer-assisted image analysis provides evidence that the CD4+ subset undergoes significant increase in organelle density after stimulation, while the CD8+ subset exhibits a post-stimulation organelle development proportional to the cell size. Each immunological subset contains two distinct patterns of cytoplasmic organization, here defined as type I (containing a few cytoplasmic organelles) and type II (with well-developed synthesis and storage organelles). On the basis of these findings cytologic parameters could be ascribed to a heterogeneous proliferation of functionally distinct T-cell clones in CD4+ and CD8+ subsets.