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Background Non-natural mortality in children and adolescents is a global public health problem that varies widely from country to country. Data on child and adolescent maltreatment are not readily available, and mortality due to violent causes is also underestimated. Methods Injury-related mortality rates (overall and by specific causes) from 2000 to 2018 in selected European countries were analysed to observe mortality patterns in children and adolescents using data from the Eurostat database. Age-standardized mortality rates per 100,000 person-years were calculated for each country. Joinpoint regression analysis with a significance level of 0.05 and 95% confidence intervals was performed for mortality trends. Results Children and adolescent mortality from non-natural causes decreased significantly in Europe from 10.48 around 2005 to 5.91 around 2015. The Eastern countries (Romania, Bulgaria, Poland, Slovakia, Czech Republic) had higher rates; while Spain, Denmark, Italy, and the United Kingdom had the lowest. Rates for European Country declined by 5.10% per year over the entire period. Larger downward trends were observed in Ireland, Spain and Portugal; smaller downward trends were observed for Eastern countries (Bulgaria, Czech Republic, Poland, Slovakia) and Finland. Among specific causes of death, the largest decreases were observed for accidental causes (-5.9%) and traffic accidents (-6.8%). Conclusions Mortality among children and adolescents due to non-natural causes has decreased significantly over the past two decades. Accidental events and transport accidents recorded the greatest decline in mortality rates, although there are still some European countries where the number of deaths among children and adolescents from non-natural causes is high. Social, cultural, and health-related reasons may explain the observed differences between countries. Key Messages • The mortality of children and adolescents due to non-natural causes has decreased significantly in the last two decades; • Substantial differences between countries were found in the trend of non-natural causes of death in children and adolescents; • Accidental events and transport accidents showed the greatest decrease in mortality; • In some European countries, there is still a high number of deaths in children and adolescents that are due to non-natural causes;

The authors present the first case, to the best of our knowledge, of a preplanned suicide by self-waterboarding. Waterboarding (WB) is a military method of torture in which water is poured into the nostrils and the mouth of a victim, to evoke the sensation of asphyxiation by drowning. The victim was a 22-year-old male student, who was found dead and naked in the bathtub. His head was covered by a soaked canvas bag, and his hands were tied with two nylon ropes and a padlock. The water jet of the showerhead was specifically directed at the victim’s head, so that the canvas bag could be soaked with water. The cause of death was defined as the combination of asphyxiation by drowning with the direct suffocation provoked by the soaked canvas bag in the context of the waterboarding practice. Finally, the authors discuss the differential diagnosis regarding the modality (suicide versus homicide) through which this case of waterboarding was performed. The case is intended to be used as source data for similar forensic cases, where a multidisciplinary approach is advisable in such complex cases.

It is commonly accepted that crime scene recovery and recording are key moments of any judicial inspection in which investigators must decide on the correct strategies to put into place. Complex outdoor scenarios, presenting partially or entirely skeletonised remains, can benefit more than others by the intervention of environmental specialists (forensic anthropologists, archaeologists, entomologists and botanists). These experts are capable of singling out, correctly recording and recovering environmental evidence that can lead to a more comprehensive reconstruction of a given criminal episode. If human remains are discovered in an outdoor scenario, the on-site presence of a botanist will guarantee a correct approach to the identification, recording and recovery of any botanical evidence. If an on-site botanist is not available, the operators must be capable of both the botanical evaluation of a scene and the implementation of correct botanical sampling protocols.The following collection of unusual case histories that aim at underlining the efficacy of forensic botany will examine the determination of post mortem or the post depositional interval, evidence for a victim’s post mortem transfer, evidence for the identification of a primary crime scene and evidence for the identification of a victim’s dismemberment site. In another two cases, one, we will illustrate the important role that forensic botany played in the discrimination between botanical material used to voluntarily conceal a victim and vegetation that had grown naturally above a disposal site, whereas the other will highlight the protocols implemented for the identification of a murder weapon.

•Cadavers in acid are difficult forensic situations.•Destruction of human bone in hydrochloric acid (HCl).•HCl 37% is more destructive than HCl 10%.•Cortical bone is more resistant than spongy bone to acid.•Recognizability as human bone tissue may be lost. The destruction of a corpse in caustic acid can cause complex forensic scenarios to deal with. Furthermore, the literature on the subject is poor, having been the few studies carried out only on animal bones. We carried out an experimental analysis on human cranial, ribs, vertebrae, and femur bone fragments. These samples were sent for dissolution by hydrochloric acid (HCl) at two different concentrations in the lab: 10% and 37%. We have performed macroscopic and microscopic histological and cytological observations at set time intervals: 3, 4, 19, 24, 48, and 72 h of immersion in acids. The purpose of the study was threefold: to investigate the temporal evolution of bone dissolution, evaluate the destructive effect of the two hydrochloric acids, and establish whether or not the human pattern of histological structure could be recognized. A more significant destructive action of HCl at 37% has been observed. In the 10% acid, the bone nature of the samples was demonstrable up to 24 h of immersion, but the human pattern of histological structure was already compromised at 19 h, being lost at 24 h. Instead, in the 37% acid, the bone nature of the sample was demonstrable only within 4 h of immersion, and the human pattern of histological structure was markedly compromised within 3 h. At 19 h of immersion, neither the recognition of the bone nor its human nature was feasible. These preliminary findings and observations may be of practical use in forensic investigations of bodies found in acidic substances, for which there is no scientific evidence to refer.

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. [Display omitted] •Postmortem microbiology (PMM) is of great forensic utility.•Thanks to PMM the number of cases without a cause of death may be reduced.•Histology allows to better assess the postmortem microbiological findings.

Background: diatoms are unicellular algae that have been used for more than a century for forensic purposes to diagnose drowning, with more or less success depending on the historical era. Although many years have passed, scientific research on diatoms has never ceased, which testifies to their enduring allure in forensics. Of course, diatom research has evolved and expanded over time, changing with the availability of new techniques and technologies. The volume of articles and their production over a period of many years has resulted in old, current, and new knowledge on diatoms being scattered over a large number of books and articles. Objectives: the purpose of this narrative literature review is, therefore, to summarize all this information and bring it together in a single work that can be useful for those who are studying diatoms and their usefulness for forensics for the first time, for those who are looking for proven methods of analysis, and finally for those who are interested in exploring new frontiers of research. Methods: a comprehensive literature search that included all studies dealing with the applications of diatoms in forensic science was performed in the most popular electronic databases. Results: traditional methods have been complemented by molecular and imaging methods and, more recently, by artificial intelligence. In addition, new biological substrates have been found for the analysis of diatoms. Conclusions: all this has led, on the one hand, to the consolidation of a whole body of knowledge on diatoms, on which this forensic analysis is still based, and, on the other hand, has opened up numerous new research directions.

The microscopic evaluation of hemorrhagic infiltrates is crucial in forensic diagnostics, but it proves challenging in corificated and mummified cadavers. In these cases, pre-treatment with rehydrating solutions is recommended, although their effects on the hemorrhagic infiltrate are not well understood. In this pilot study, we microscopically investigated the effect of two different rehydrating solutions—Sandison’s solution and fabric softener—on well-preserved human cadaveric skin samples taken from areas affected by an ecchymotic lesion, comparing them with direct fixation in formalin. Specifically, we examined the topographic distribution of the hemorrhagic infiltrate in each layer of the skin by assigning a semi-quantitative score, conducted mutual comparisons, and performed statistical analysis. Histologically, compared to direct fixation in formalin, a slight and statistically non-significant reduction in the hemorrhagic infiltrate was observed in samples pre-treated with fabric softener. On the other hand, a more pronounced and statistically significant decrease in scores was observed in samples pre-treated with Sandison’s solution. This effect is likely due to the fact that Sandison’s solution, due to its components, exerts an osmotic effect, partially inducing osmotic lysis of red blood cells. Overall, extensive areas of hemorrhagic infiltrates were preserved, although to a lesser extent, while smaller foci were markedly reduced, sometimes even disappearing. The findings suggest that Sandison’s solution has a detrimental effect on cutaneous hemorrhagic infiltrates, emphasizing the importance of being cautious and conducting dual sampling, using both formalin and a rehydrating solution, for forensic examination of mummified or corificated skin samples.

The discovery of new methods for determining the post-mortem interval is of significant forensic interest. Mass spectrometry has enhanced the accuracy of assessing post-mortem protein decay, with skeletal muscle being the most studied substrate due to its intrinsic properties of postmortem decay. In this pilot study, human skeletal muscle tissue (iliopsoas) was harvested and allowed to decay under controlled temperature and humidity conditions at predetermined intervals. The samples were analyzed using mass spectrometry proteomics for both qualitative and quantitative evaluation of proteins and peptides. Candidate proteins were validated through immunoblotting. The results were significant, identifying several proteins that could aid in estimating the post-mortem interval. Notably, PLIN4, MYOZ2, SYNPO2, and BAG3 were validated by immunoblotting over a broader range of experimental points and temperatures. Furthermore, human results were compared with animal muscle samples from a previous study, revealing similarities in decomposition kinetics. This analysis of human samples marks a step forward in the potential forensic application of proteomic evaluation by mass spectrometry. •Proteomic approach seems to be beneficial for PMI estimation.•Mass spectrometry constitutes a significant advancement in the analysis of protein markers for PMI.•The proteins with better PMI kinetics appear to be MYOZ2, HNRNPA2B1 and PDLIM1.

Mass spectrometry (MS)-based proteomics has recently attracted the attention from forensic pathologists. This work is the first report of the development of a shotgun bottom-up proteomic approach based on rapid protein extraction and nano-liquid chromatography/high-resolution mass spectrometry applied to full-thickness human skin for the differential analysis of normal and ecchymotic tissues to identify new biomarkers for bruise characterization and dating. We identified around 2000 proteins from each pooled extract. The method showed excellent precision on independent replicates, with Pearson correlation coefficients always higher than 95%. Glycophorin A, a known biomarker of vital wounds from immunochemical studies, was identified only in ecchymotic tissues, as confirmed by Western blotting analysis. This finding suggests that this protein can be used as a MS-detectable biomarker of wound vitality. By focusing on skin samples from individuals with known wound dating, besides Glycophorin A, other proteins differentially expressed in ecchymotic samples and dependant on wound age were identified, although further analysis on larger datasets are needed to validate these findings. This study paves the way for an in-depth investigation of the potential of MS-based techniques for wound examination in forensic pathology, overcoming the limitations of immunochemical assays.

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.