Pietro Zuccarello

Distinguishing antemortem from postmortem thermal injury remains one of the most challenging tasks in forensic pathology, particularly in cases involving rapid death, extensive charring, or advanced tissue degradation. Classical macroscopic and histological findings provide essential information but often lack sensitivity or specificity when considered in isolation. This review critically examines current evidence on burn vitality markers, with a primary focus on histological and immunohistochemical findings supported by human forensic studies, while contextualizing molecular and microRNA-based (miRNA-based) approaches as largely experimental and translational. Established indicators in forensic casework include erythrodiapedesis, fibrin deposition, inflammatory cell infiltration, and glycophorin-confirmed hemorrhage, together with selected immunohistochemical markers such as HSP70 (Heat Shock Protein 70) and aquaporin-3 (AQP3), which provide supportive evidence of tissue viability. In contrast, molecular biomarkers, such as cytokine gene expression, oxidative stress products, mitochondrial DNA (mtDNA) release, and miRNA signatures, offer mechanistic insight into early cellular responses to thermal injury but currently lack sufficient forensic validation in human autopsy-based studies to support routine medico-legal interpretation. The available evidence indicates that histological and selected immunohistochemical findings constitute the most reliable basis for burn vitality assessment, whereas molecular approaches should be interpreted cautiously as adjunctive and investigational. Diagnostic conclusions therefore rely on the integration of multiple independent findings, evaluated in relation to survival time, tissue preservation, and postmortem artifacts. Overall, this review emphasizes that burn vitality determination requires a cautious, multimodal, and context-driven approach, grounded in validated forensic evidence and supported, where appropriate, by complementary experimental data. Further progress in this field will depend on the systematic validation of emerging markers in human forensic autopsy cohorts.

Background/Objectives: Establishing drowning as the cause of death remains one of the most challenging tasks in forensic pathology, particularly when classical autopsy findings are absent, nonspecific, or compromised by decomposition. Molecular biomarkers may provide objective adjunctive evidence in such cases. This study investigated the diagnostic utility of Aquaporin-5 (AQP5) and Surfactant Protein A2 (SP-A2) measured in renal tissue as potential biomarkers of freshwater drowning and evaluated the influence of different freshwater environments on their expression. Methods: A controlled experimental study was performed using 29 adult Wistar rats allocated to freshwater drowning groups (Boračko Lake, n = 11; Jablaničko Lake, n = 11) and non-drowning control groups subjected to mechanical asphyxia followed by one hour of postmortem immersion (n = 7). Kidney tissues were collected within a maximum of 5 min after death and analyzed for AQP5 and SP-A2 concentrations using enzyme-linked immunosorbent assay (ELISA). Group differences were evaluated using Kruskal–Wallis and Dunn’s post hoc tests, while effect sizes were estimated using Cliff’s delta. Receiver operating characteristic (ROC) curve analysis was performed to assess the diagnostic performance of each biomarker in distinguishing drowning from non-drowning deaths. Results: AQP5 concentrations differed significantly among groups (χ² = 16.05, p = 0.00033), with large effect sizes observed in all pairwise comparisons. However, diagnostic performance was poor, as ROC analysis demonstrated no significant discrimination between drowning and non-drowning cases (AUC = 0.534, 95% CI: 0.314–0.754; p = 0.762). In contrast, SP-A2 showed significant intergroup differences (χ² = 6.69, p = 0.035) and moderate diagnostic accuracy for drowning identification (AUC = 0.752, 95% CI: 0.567–0.937; p = 0.008), with a sensitivity of 77.2% and specificity of 81.8% at the optimal threshold. Notably, AQP5 expression appeared suggestive of environmental modulation, whereas SP-A2 demonstrated greater relevance to the drowning process itself. Conclusions: Renal SP-A2 showed moderate diagnostic utility as a supportive biomarker for drowning, whereas AQP5 lacked sufficient discriminatory power for cause-of-death determination. The marked variability of AQP5 across freshwater environments suggests that it reflects environmental modulation rather than drowning-specific biological responses. Although neither marker is suitable as a standalone diagnostic tool, renal SP-A2 may represent a useful adjunct in forensic drowning investigations, particularly when pulmonary tissue is unavailable or unsuitable for analysis. These findings should be considered preliminary and applicable only to freshwater drowning conditions, as saltwater drowning was not investigated in the present study. Further validation in human forensic casework and larger experimental cohorts is warranted.

Background/Objectives: DNA-based identification of degraded human remains represents a major challenge in forensic science, particularly in cases involving burned, fragmented, or commingled bodies. Advances in forensic genetics have expanded the analytical capabilities for such samples; however, the effectiveness of different approaches and their integration within Disaster Victim Identification (DVI) workflows remain heterogeneous. This systematic review aims to critically evaluate current evidence on DNA-based identification of degraded remains, focusing on methodological strategies, emerging genomic technologies, and DVI applications, while integrating laboratory evidence and operational forensic practice into a structured analytical framework. Methods: A systematic literature search was conducted in Scopus and Web of Science from database inception to 5 June 2026, following PRISMA 2020 guidelines. Eligible studies included original research addressing DNA analysis of degraded, thermally altered, or highly compromised human remains in forensic or DVI contexts. After a multistep screening process involving title/abstract and full-text evaluation, 37 studies were included. Data were extracted and organized into three thematic categories: (i) core DNA analysis, (ii) advanced molecular technologies, and (iii) DVI case applications. Results: The findings demonstrate that DNA recovery from degraded remains is influenced by thermal exposure, tissue type, and sampling strategy. Teeth and dense cortical bone consistently provide higher DNA yield. While autosomal STR profiling remains the primary analytical approach, its limitations in highly degraded samples are mitigated through the complementary use of mitochondrial DNA (mtDNA), Y-chromosome STRs (Y-STRs), and SNP markers, together with advanced sequencing technologies such as massively parallel sequencing (MPS). Emerging technologies, including rapid DNA systems and predictive models based on macroscopic indicators, significantly enhance efficiency and success rates. DVI studies report identification rates exceeding 90–95% when multidisciplinary and structured workflows are applied. The evidence further supports a flexible triage-based analytical strategy, in which marker selection is guided by tissue preservation and degradation level. Conclusions: DNA-based identification of degraded human remains has evolved into an adaptive, multi-level forensic process. Successful outcomes rely on the integration of optimized sampling, hierarchical genetic analysis, and coordinated DVI strategies. The findings support a triage-based framework that links tissue selection, degradation assessment, and analytical methodology to maximize identification success. Future developments should focus on predictive models, advanced genomic tools, and standardized workflows to further improve identification in challenging forensic scenarios.

Evidence indicates that micro and nanoplastics (MNPs) may act as endocrine disruptors, promote neuroinflammation, and interfere with treatments. This study investigated the association between internal MNP exposure and major depressive disorder (MDD) through a comparative biomonitoring design conducted between July 2021 and June 2022. Blood and urine MNPs were quantified using patented extraction protocol and identified by SEM-EDX, while polymer characterization by PyGC/MS R,Scitalopram concentrations were measured via UPLCMS/MS. Thirty individuals with MDD and thirty controls were enrolled. Cases exhibited higher median blood MNP levels than controls (11,570 vs 6,186 p/mL), a pattern confirmed in sexstratified analyses for both men and women (13,139 vs 11,570 p/mL). Among controls, females showed higher levels than males (9,028 vs 4,138 p/mL). Urinary MNPs were more frequently detectable and substantially elevated in cases (152 vs 0 p/mL). Particle size did not differ by group or sex. Participants with blood MNP levels ≥8,780 p/mL had higher estimated odds of MDD (adjusted OR 4.70; 95% CI 1.17–19.0), whereas older age was associated with lower odds. Polyethylene and polypropylene predominated in both matrices. No differences in blood MNPs emerged between adherent and nonadherent citalopram users. Findings suggest a potential link between MNP burden and MDD.

Evidence indicates that micro and nanoplastics (MNPs) may act as endocrine disruptors, promote neuroinflammation, and interfere with treatments. This study investigated the association between internal MNP exposure and major depressive disorder (MDD) through a comparative biomonitoring design conducted between July 2021 and June 2022. Blood and urine MNPs were quantified using patented extraction protocol and identified by SEM-EDX, while polymer characterization by PyGC/MS R,Scitalopram concentrations were measured via UPLCMS/MS. Thirty individuals with MDD and thirty controls were enrolled. Cases exhibited higher median blood MNP levels than controls (11,570 vs 6,186 p/mL), a pattern confirmed in sexstratified analyses for both men and women (13,139 vs 11,570 p/mL). Among controls, females showed higher levels than males (9,028 vs 4,138 p/mL). Urinary MNPs were more frequently detectable and substantially elevated in cases (152 vs 0 p/mL). Particle size did not differ by group or sex. Participants with blood MNP levels ≥8,780 p/mL had higher estimated odds of MDD (adjusted OR 4.70; 95% CI 1.17–19.0), whereas older age was associated with lower odds. Polyethylene and polypropylene predominated in both matrices. No differences in blood MNPs emerged between adherent and nonadherent citalopram users. Findings suggest a potential link between MNP burden and MDD.

Background: Postmortem redistribution (PMR) is a major challenge in forensic toxicology, particularly when drug concentrations are measured in central blood rather than peripheral blood. Although peripheral blood remains the preferred specimen for postmortem interpretation, it is not always available in routine forensic practice. Methods: This retrospective descriptive study evaluated postmortem central blood drug findings from judicial autopsies analyzed at the Forensic Toxicology Laboratory of the University of Catania between 2017 and 2024. Approximately, 700 forensic toxicological investigations were reviewed. Overall, 52 therapeutic drugs were identified, resulting in a final dataset of 313 individual drug findings. For each finding, central blood concentrations were compared with published reference ranges and cautiously classified as normal/therapeutic, high but non-toxic, toxic, or potentially lethal. Results: Most findings were classified as normal/therapeutic (84.3%), while 28 (8.9%) were high but non-toxic, 15 (4.8%) toxic, and 6 (1.9%) potentially lethal. The distribution remained broadly consistent in sensitivity analyses restricted to drugs represented by more than 3, 5, or 8 findings. In cases with toxic or potentially lethal findings and available paired peripheral blood data, observed C/P ratios were moderate, ranging from 1.07 to 2.15. Conclusions: These findings do not indicate that central blood can replace peripheral blood or that PMR can be disregarded. However, they suggest that central blood drug concentrations are not intrinsically uninterpretable and may retain practical forensic value when interpreted cautiously within the broader medico-legal context. Central blood results should not be used in isolation to diagnose poisoning, estimate the ingested dose, or reconstruct exact ante mortem concentrations.

Sudden cardiac death (SCD) in patients with schizophrenia treated with antipsychotic drugs (APDs) has become a significant concern, with increasing incidence, particularly among young adults. This multicenter study aimed to investigate the cardiac effects of APDs in a cohort of patients who died from SCD. A retrospective analysis was conducted on seven schizophrenic patients, aged 25 to 39 years, who died from SCD while receiving antipsychotic therapy. Patients with cardiovascular risk factors, substance abuse at the time of death, or advanced age were excluded. Autopsies focused on cardiac tissue, and histological and toxicological analyses were performed. The findings revealed structural cardiac changes, including myocardial fibrosis, myofiber disruption, and contraction band necrosis—features consistent with arrhythmic SCD. Toxicological results confirmed APD use in all cases, with polypharmacy patterns commonly observed. The study suggests that APDs can induce significant cardiac alterations that may predispose patients to SCD, even in the absence of pre-existing cardiac disease. These findings underscore the need for careful cardiac monitoring in patients with schizophrenia treated with APDs, particularly those without traditional cardiovascular risk factors. Further research, including molecular autopsy and genetic analysis, is necessary to better understand genetic predispositions and the role of APDs in SCD.

Background/Objectives: I Paliperidone is an antipsychotic recently added into the market in various formulations. There are few data about safety and on therapeutic, toxic, or lethal blood concentrations. Currently, the published analytical methods are often applied to serum or plasma that are not obtained from cadaveric blood. Alternatively, aliquots of high volume of whole blood are used, but often in forensic investigations using samples at very small quantities. The aims of the present study were (a) to develop an analytical method to detect and quantify paliperidone in whole blood using only a small sample volume (10 µL) and (b) to summarize data on post-mortem blood analysis obtained from authentic autopsy cases. Methods: Method validation was carried out on 10 µL of whole blood, extracted by LLE and analyzed by LC-MS. Paliperidone concentrations obtained from blood analysis of 16 authentic autopsy cases were reported. Results: The method showed a good linearity and sensitivity, a normal distribution, the absence of anomalous values, an interday RSD% always less than 10%, and an 80–120% recovery, as required by AAFS guidelines. Femoral blood concentrations obtained from authentic autopsy cases ranged between 23.4 and 146.9 ng/mL. Conclusions: This method is to be used properly in all cases where it is necessary (a) to monitor the therapeutic adherence of patients, (b) to establish the psycho-physical conditions of the treated subject at the time of the death, and (c) to ascertain if the drug may have played a causal role in the obitus. This study reported the first data obtained from post-mortem investigation of subjects treated with paliperidone LAI. Cadaveric blood concentrations could be higher than ante-mortem reference values due to post-mortem redistribution.

Food aspiration is one of the major health risks for elderly people in nursing homes which could lead to death. Moreover, misconducts in pharmacotherapy may represent a potential risk of adverse drug reactions. It is reported here the toxicological evaluation of a combined death by food aspiration and acute escitalopram intoxication of a psychiatric subject, occurred in a nursing home. An 89-year-old man, suffering from dysphagia and Alzheimer’s, was resident in a nursing home. He was fed with a liquid diet administered directly in mouth using a syringe. The man was also being treated with escitalopram 10 mg tablet. One evening, after receiving the meal in the usual way, the man complained of sudden illness. Carried to the emergency room, the man died about 3 h later with a diagnosis of cardiogenic shock subsequentially to ab ingestis. The histological findings revealed the presence of exogenous material, probably food, up to the finest bronchial branches. The toxicological examination revealed the presence of escitalopram and its main metabolite, desmethylcitalopram: in the blood 1972 ng/ml and 285 ng/ml, in the brain 4657 ng/g and 1025 ng/g, in the gastric content 2317 ng/g and 423 ng/g, in the lung 21,771 ng/g and 468 ng/g, respectively. The bad practice of the nurses to dissolve the escitalopram tablet in the liquefied food and to administer the therapy with a syringe directly into the mouth emerged thanks this investigation. Following food aspiration, escitalopram was absorbed by inhalation route, reaching high concentrations in blood and tissues. The death occurred due to a combined mechanism between food aspiration and the escitalopram toxic action.